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									                                  Research Summaries Book
                                  ENDO 2003
                             th
                The 85 Annual Meeting of The Endocrine Society
                                June 19–22, 2003
            Pennsylvania Convention Center, Philadelphia, Pennsylvania
                                      Press Room: 304
            Hours: Thursday–Saturday, 7:30 a.m.–5:00 p.m.; Sunday, 8:00 a.m. – noon

                                  HOW TO USE THIS BOOK
The ENDO 2003Research Summaries Book has been designed to facilitate media coverage of the 85th
Annual Meeting of The Endocrine Society. The highlighted abstracts contained in this book have been
identified as those that would be of particular interest to journalists. They were selected from among
nearly 2,500 abstracts that will be presented at ENDO 2003.

Each abstract in this book is presented in its original scientific form, just as it appears in the Program and
Abstracts Book. A news summary of the research, which was prepared in coordination with the research
teams, follows each abstract. The summaries include additional background information on the area of
research and, in many cases, the source of funding for the research. The original abstracts appear on the
left-hand pages and the news summaries on the facing right-hand pages.

The abstracts are divided into general topic areas, as denoted in the table of contents, which follows this
page. Hot Topics and Clinical Trials Symposia, the first section of this book, features results of up-to-the-
minute research and the latest clinical trials. At the end of the book, there are indices of abstracts in
numerical order, by author, date of presentation and by research funding. In addition, this book includes a
special compilation of abstracts related to the focus of ENDO 2003,“Cardiovascular Endocrinology.”
These abstracts are listed in the “Cardiovascular Abstracts” index at the back of the book and are marked
in the indices with the symbol: ♥

The full ENDO 2003 program is available on The Endocrine Society’s Web site through the On-line
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       ENDO 2003 Embargo Policy


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 embargoed until 6:00 a.m. (EST) on Thursday, June 19, 2003. The
 Endocrine Society reserves the right to lift the embargo on specific
abstracts that are selected for promotion prior to the start of ENDO
                                 2003.




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                                   Table of Contents


1.    Clinical Trials/Hot Topics                       pp. 1–6
2.    Aging                                            pp. 7–14
3.    Androgen Disorders                               pp. 15–26
4.    Cardiovascular Endocrinology                     pp. 27–38
5.    Diabetes                                         pp. 39–52
6.    Effects of Hormones on Cancer                    pp. 53–56
7.    Gene Expression                                  pp. 57–60
8.    Glucocorticoids                                  pp. 61–64
9.    Growth Hormone                                   pp. 65–80
10.   Nuclear Receptors                                pp. 81–84
11.   Obesity                                          pp. 85–112
12.   Osteoporosis                                     pp. 113–134
13.   Pediatric Endocrinology                          pp. 135–148
14.   Reproduction                                     pp. 149–182
15.   Steroid Hormone Action/Biosynthesis              pp. 183–194
16.   Thyroid                                          pp. 195–206
17.   Indices
         Day of Presentation                          pp. 207–210
         Author                                       pp. 211–213
         Subject/Headline                             pp. 214–218
         Abstract Number                              pp. 219–221
         Funding Source                               pp. 222–224
Clinical Trials/Hot Topics
                     ENDO 2003 Clinical Trials Symposium
                     Friday, June 20, 2003
                     9:30–11:00 a.m.
                     Room 204A, Pennsylvania Convention Center

Each year, the Clinical Trials Symposium includes announcements of groundbreaking clinical
trials that have never been presented at a meeting. The ENDO 2003 Clinical Trials Symposium
topics will include parathyroid hormone treatments, testosterone therapy, DHEA and growth
hormone deficiency.


                   Clinical Trials Symposium Presentations:


    S41-1 “Blood pressure Control in the Veterans Affairs Diabetes Trial
     (VADT)”
          Presented By Dr. Robert J. Anderson, Veterans Administration
          Medical Center (VAMC)


    S41-2 “Efficacy of Triiodothyronine (T3) Addition to Paroxetine in
     Major Depressive Disorder: A Randomized Clinical Trial”
          Presented By Dr. Eric Fliers, Academic Medical Center, University of
          Amsterdam


    S41-3 “Effects of Growth Hormone Administration (by
     Individually Tailored Dose titration) and/or Testosterone on
     Body Composition, Physical Performance and Quality of life in
     Healthy Elderly Men”
           Presented By Dr. Manthos G. Giannoulis, King’s College, St. Tomas
           Hospital




                                             1
2
               ENDO 2003 Hot Topics Symposium
               Saturday, June 21, 2003
               9:30–11:00 a.m.
               Room 204A, Pennsylvania Convention Center


The Hot Topics presentations focus on the most pivotal developments in
endocrinology. The ENDO 2003 Hot Topics presentations will cover blood
pressure, growth hormones and diabetes.


                Hot Topics Symposium Presentations:


   S41-1 “LRp5 and Bone Density”
         Presented By Dr. Robert Recker, Creighton University, Omaha,
         Nebraska

   S41-2 “Ghrelin and the Regulation of Appetite and Body Weight”
         Presented By Dr. David Cummings, University of Washington,
         Seattle, Washington

   S41-3 “PTH Type 1 Receptor Mutation in Enchondromas
         Presented By Dr. Benjamin Alman, Hospital for Sick Children,
         Toronto, Ontario




                                    3
4
                                                                                                    Clinical Trials
S19-1
Blood Pressure Control in the Veterans Affairs Diabetes Trial (VADT).
Robert J Anderson*1, Carlos Abraira2, William C Duckworth3, Madeline McCarren4, Mark B Zimering5, the VADT
Trial Group4. 1Endocrinology, VA Med Ctr, Omaha, NE; 2Res Svc, Miami VA Med Ctr, Miami, FL;
3
  Endocrinology, Carl T. Hayden VA Med Ctr, Phoenix, AZ; 4Coop Studies Coordinating Ctr, Hines VAMC,
Chicago, IL; 5Endocrinology, VA Med Ctr, Lyons, NJ.

It is not known whether intensive glycemic control is beneficial in delaying cardiovascular (CV) disease in type 2
diabetes. Blood pressure (BP) control is consistently correlated with CV outcomes in type 2 diabetes. In the UKPDS
it was twice as effective in preventing microvascular endpoints as was glycemic treatment. The VADT is a 20 –
center, 1700 patient prospective, randomized study of Intensive vs. Standard glycemic treatment in patients with
type 2 diabetes responding sub-optimally to maximum oral agents or insulin. The main objective is to address the
risk/benefit ratio of intensive glycemic control for up to 7 years on CV outcomes in patients with advanced type 2
diabetes. Other objectives include the assessment of effects on microvascular and neurological complications,
cognitive function, quality of life and cost effectiveness. BP, lipids, diet and lifestyle are treated identically in both
arms. In 1250 randomized subjects in this ongoing trial, mean age was 61, BMI 31, HbA1c 9.4%, and diabetes
diagnosis 11 years. Forty percent of patients had prior CV events. At entry, mean BP was 131/77 mmHg. All
entering the trial with new or treated high BP are given stepped treatment, beginning with angiotensin-converting
enzyme inhibitors or angiotensin II receptor blocking agents, with the addition of the following agents as needed:
low dose diuretics, cardioselective beta blockers or long acting calcium channel blockers, and clonidine. The cohort
of 771 patients at 6 months into the trial had a mean blood pressure of 126/74 mmHg, with no differences between
the standard and intensive glycemic treatment arms. Eighty-one percent of patients were treated with rosiglitazone
(8 mg in the Intensive arm vs. 4 mg in the Standard arm), and all patients were treated either with other oral agents,
insulin, or combinations of both therapies. Conclusion: The cohort entering VADT had near optimal mean BP,
which was further reduced within 6 months with additional BP treatment. By improving blood pressure control in an
identical manner in both glycemic arms, the VADT can exclude the effect of blood pressure differences in CV
events, and may reduce macrovascular complications during the trial.
           Supported by the Cooperative Studies Program of the Department of Veterans Affairs Research and
Development Service; also by the American Diabetes Association, National Eye Institute, GlaxoSmithKline, Novo
Nordisk, Aventis, Roche Diagnostic, and KOS Pharmaceuticals.

CLINICAL SYMPOSIUM: Clincial Trials, 6/20/2003, (9:30 AM – 11:00 AM); Location Ballroom A




                                                            1
                                                                                 Clinical Trials

S19-1 News Summary
Improving blood pressure in diabetics reduces other health problems

Improving blood pressure in diabetics reduces complications from eye, kidney and
cardiovascular problems, according to preliminary results from an ongoing clinical trial being
presented on Friday, June 20, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

More than 16 million Americans have diabetes, a condition characterized by high blood sugar
levels. At least 90 percent of diabetics have type-2 diabetes, caused when the body cannot
produce enough of a hormone called insulin – which helps remove excess sugar from the blood –
or when the body cannot use insulin properly. The majority of diabetic-related hospitalizations
and deaths are due to cardiovascular complications, such as heart attack and stroke. The
likelihood of drug side effects increases and insulin therapy can have adverse effects on people
who are older, overweight and in poor cardiac health. It has not been proven that control of
blood glucose will prevent cardiovascular complications. However, control of blood pressure
reduces these complications, and it has also been more than twice as effective as blood glucose
control in reducing eye and kidney complications in type-2 diabetes.

The Veterans Affairs Diabetes Trial (VADT), led by Dr. Robert J. Anderson, is examining the
effect of glucose control on cardiovascular events, in hope of determining the proper level of
treatment to decrease complications. Considering the number of Americans with diabetes and
the estimated $130 billion per year diabetes costs the nation, finding a way to reduce
complications would have a major impact on health care in the United States.

The VADT includes 1700 patients from 20 different VA hospitals with type-2 diabetes, who
were poorly controlled with their current treatment. Enrollment has been completed. The trial
started in December 2000 and is scheduled to continue for 7 years. The participants were
divided into two groups; one group receives a standard combination of oral agents and insulin
while the other receives a more intensive drug therapy with similar medications. All patients in
both groups are treated identically with respect to blood pressure, cholesterol, aspirin use, diet
and lifestyle.

The first 1,250 randomized patients were, on average, 61 years old, obese, poorly controlled and
had known diabetes for 11 years. Forty percent reported earlier cardiovascular problems. Their
average blood pressure at entry was considerably better than earlier reports in other diabetes
populations. By six months, the data showed that average blood pressure was even lower and
within current diabetes treatment targets, without difference between the two groups. All patients
with known high blood pressure were treated with a similar combination of blood pressure pills.

The trial is supported by the Cooperative Studies Program of the Department of Veterans Affairs
Research and Development Service, American Diabetes Association; National Eye Institute,
GlaxoSmithKline; Novo Nordisk, Aventis, Roche Diagnostic and KOS Pharmaceuticals.
                                              # # #


                                                 2
                                                                                                Clinical Trials
S19-2
Efficacy of triiodothyronine (T3) addition to paroxetine in major depressive disorder: a randomized clinical
trial
Eric Fliers*1, Bente C Appelhof1, Jantien P Brouwer1, Witte J Hoogendijk2, Jochanan Huyser1, Aart H Schene1, Jan
GP Tijssen1, Richard Van Dyck2, Wilmar M Wiersinga1. 1Depts of Endocrinology and Metab, Psychiatry and
Cardiol, Acad Med Ctr, Univ of Amsterdam, 1105 AZ Amsterdam, Netherlands; 2Dept of Psychiatry, Free Univ,
1075 BG Amsterdam, Netherlands.

Background. The addition of triiodothyronine (T3) to antidepressants has been advocated as an augmentation
strategy in the treatment of major depressive disorder. However, evidence has been derived exclusively from
relatively small clinical studies. In addition, the efficacy of T3 co-medication has been studied only in combination
with tricyclic antidepressants.
Aim. To investigate the efficacy of T3 addition to a selective serotonin reuptake inhibitor (SSRI) in patients with
major depressive disorder.
Method. We randomly assigned 113 men and women with major depressive disorder and a baseline 17-item
Hamilton Rating Scale for Depression (HRSD) score 16 to 8 weeks of double-blind outpatient treatment with low
dose (25 g), high dose (50 g) or placebo in addition to paroxetine 30 mg daily. We excluded patients with overt
hypo- or hyperthyroidism. Response was defined as a reduction of the HRSD score 50% and remission as a HRSD
score 8 at endpoint. Secondary outcome measures included the Montgomery Asberg Depression Rating Scale
(MADRS) and the Beck Depression Inventory (BDI).
Results. Hundred and six patients started treatment and were included in the final analysis of response. Response
rate after eight weeks was 46% in all three treatment arms (p=0.99). Remission rate was 32% in both T3 arms and
36% in the placebo group (p=0.92) (Table 1). None of the secondary outcome measures showed significant
differences between the treatment groups. The mean time interval to response did not differ between the treatment
groups (5.3, 5.9 and 6.3 weeks in the placebo, 25 and 50 microgram T3 group, respectively). Patients on T3 addition
reported significantly more side effects than patients on placebo addition. Nevertheless, attrition rates were similar
in the three groups.
Conclusion. These results do not support a role for T3 addition to SSRI in the treatment of major depressive
disorder.
                                          Table 1. Primary outcome
                              placebo (n=50) 25 g T3 (n=28) 50 g T3 (n=28) total (n=106) p-value
HRSD
Baseline (mean±SD)            20.7±3.2         20.7±3.3          20.8±3.7          20.7±3.3      0.98
End of treatment (mean±SD)    11.5±6.6         11.2±5.7          12.7±5.8          11.7±6.1      0.60
Response (n; %)               23; 46.0%        13; 46.4%         13; 46.4%         49; 46.2%     0.99
Remission (n; %)              18; 36.0%        9; 32.1%          9; 32.1%          36; 34.0%     0.92


CLINICAL SYMPOSIUM: Clincial Trials, 6/20/2003, (9:30 AM – 11:00 AM); Location Ballroom A




                                                           3
                                                                                  Clinical Trials
S19-2 News Summary
Thyroid hormone T3 provides more problems than benefits in treating depression, despite
current treatment guidelines for its use

Despite current recommendations, the thyroid hormone T3, when taken along with an
antidepressant, provides no extra benefits for patients with major depression, according to the
first clinical trial testing its use. The data are being presented on Friday, June 20, at The
Endocrine Society’s 85th Annual Meeting in Philadelphia. In addition, the study found that those
who took T3 actually had more side effects.

An important part of the therapy for patients with major depression is treatment with
antidepressant drugs. Most physicians prefer the so-called selective serotonin reuptake inhibitors
(SSRI) because they have only a few side effects. However, a large proportion of depressed
patients – some 40 percent – do not respond to this therapy. Official guidelines of the American
Psychiatric Association (APA) mention the possibility of adding the thyroid hormone T3, or
triiodothyronine, to antidepressants so as to enhance therapeutic effect. However, there have
been no clinical trials investigating if the combination of SSRI and T3 is more effective than
SSRI alone.

Because of this void of data, Dr. Eric Fliers and colleagues in Amsterdam designed a
randomized, double-blind, placebo-controlled clinical trial to assess the efficacy of T3 addition
to the SSRI paroxetine in patients with major depression. In the study, 106 patients were
randomized to low-dose thyroid hormone, high-dose thyroid hormone or placebo in addition to
paroxetine. They observed a positive response to treatment according to pre-defined criteria in
46 percent of the patients, irrespective of whether they had placebo, low-dose or high-dose T3
added to the SSRI. The addition of T3 did not enhance or accelerate response. Patients treated
with T3 reported more side effects.

The researchers note that this study is likely to impact psychiatric patient care, and it could lead
to adaptations of the APA guidelines for the treatment of major depression. In addition, these
results may prompt a review of the use of T3 in other clinical situations, such as hypothyroidism
and critical illness.

The trial was supported by the Board of the Academic Medical Center of the University of
Amsterdam.

                                               # # #




                                                 4
                                                                                                Clinical Trials
S19-3
Effects of Growth Hormone Administration (by Individually Tailored Dose Titration) and/or Testosterone on
Body Composition, Physical Performance and Quality of Life in Healthy Elderly Men
Manthos G Giannoulis*1, Carolyn V McMillan3, Clare Bradley3, Louise Breen1, Margot Umpleby1, Peter H
Sonksen1, Finbarr Martin2. 1Div of Med,Dept of Diabetes & Endocrinology,, King’s Coll, London, United
Kingdom; 2Elderly Care & Acute Med, King’s Coll, London, United Kingdom; 3Hlth Psych Res, Royal Holloway,
Univ of London, Egham, Surrey, United Kingdom.

The aim of the study was to compare the effects of GH, and/or Testosterone (T) on body composition, muscle
strength, aerobic capacity (VO 2 max) and quality of life (QoL). There is some evidence of synergistic interaction
between GH and T.We hypothesised that the co-administration of GH at doses titrated to achieve a target IGF-I
normal but not high for younger adults with T, would enable effectiveness whilst avoiding undesirable side effects.
Eighty healthy elderly men [mean (SEM) age 70(0.3) and BMI 26.4(0.3)], were randomised to placebo GH and
placebo T (P), GH and placebo T (GH), T and placebo GH (T) or GH and T (GHT) for a six month period in a
double blinded trial. The starting dose of GH was 0.1mg/day. Serum IGF-I levels were reviewed at week 2, 4, 6, and
8 and the dose of GH was adjusted accordingly. T was given as transdermal patches. We measured lean and fat mass
by dual-energy x-ray absorbsiometry, knee extensor and flexor strengths by dynamometer. QoL was assessed by a
new questionnaire developed for this study, the A-RHDQoL, measuring perceived impact of age-related hormone
decline in QoL and perceived health by SF-36 . All measurements were performed blind at 0 and 6 months.
Serum IGF-I levels increased significantly after GH and GHT (p<0.0001)[mean (SEM) P= 12.1(0.7), GH=25.3(1.4),
T=13(0.6) and GHT=27.4(1.8) nmol/l]. Similarly T levels increased after T and GHT but reached statistical
significance only in the T group (p<0.04) [P=15(1.1), GH=16.9 (1.3), T=24.8(3.3) and GHT=20.6(2.8) nmol/l].
Total lean mass increased when compared to placebo after GH and GHT (p<0.008) but not after T, [GH=1.7(0.6),
T=0.9(0.5) and GHT=1.6(0.6) Kg]. Percentage body fat decreased compared to placebo after GH (p< 0.03) and
GHT (p<0.008). However, fat total mass decreased significantly only after GHT (p<0.03) . VO2 max increased
compared to placebo only after GHT p<0.001. In addition, GHT, resulted in a significant increase in strength of
knee extension at angular velocity of 90° and knee flexion at 120° (p< 0.001) out of a total six observations.No
significant changes occurred in QoL.There were no major side effects during the 6months trial.
We conclude that the co-administration of GH and T results in greater potentially beneficial changes than either GH
or T alone, and that, using the GH dose titration method, this can be achieved without the significant adverse effects
reported in previous studies.

CLINICAL SYMPOSIUM: Clincial Trials, 6/20/2003, (9:30 AM – 11:00 AM); Location Ballroom A




                                                          5
                                                                                Clinical Trials
S19-3 News Summary
Growth hormone and testosterone together may counteract effects of aging on the body

To counter the effects of aging on the body, patients who take both growth hormone (GH) and
testosterone see greater benefits than those who take either GH or testosterone alone, according
to a clinical trial being presented on Friday, June 20, at The Endocrine Society’s 85th Annual
Meeting in Philadelphia. In addition, these benefits can be achieved without significant adverse
effects, which have been reported in other studies, by increasing the GH dose gradually.

GH is a hormone secreted by the pituitary gland. Its most important functions are promoting
growth in children and building up muscle and breaking down fat in adults. Testosterone, the
main male sex hormone, has similar actions to GH in that it helps build up muscle and break
down fat. It also increases bone strength and improves libido and erections.

Declining levels of GH and testosterone secretion in older people have been cited as possible
causes of the detrimental body changes – such as low muscle mass, low muscle strength and
increased body fat – that accompany aging. Most previous studies examining the use of GH and
testosterone in the elderly used the hormones alone. They either failed to show convincing
results when lower doses were used or were hampered with side effects from higher doses,
specifically with GH.

Dr. Manthos G. Giannoulis and colleagues in the United Kingdom hypothesized that combined
near-normal supplementation with GH and testosterone would reverse some of these changes in
the aging process and elicit few side effects. Eighty healthy elderly men with low testosterone
and GH levels, compared to young adults, were recruited for the study. They were randomized
into four groups: 1) Placebo GH and placebo testosterone, 2) GH and placebo testosterone, 3)
Testosterone and placebo GH and 4) GH and testosterone. Each group was treated for a six-
month period. GH replacement started at a low dose, and the dose was increased slowly to
minimize side effects.

Measurements also were taken for whole body muscle and fat by DEXA scan; fat around the
waist and muscle in the middle thigh by CT scan; muscle strength in the leg, at the knee joint
level in the forward and backward movement; exercise capacity by exercising on a bike and
quality of life and sexual mood.

Subjects in the GH alone and GH and testosterone groups had increased muscle mass and
reduced body fat after six months. However, only the group taking both GH and testosterone
improved their exercise capacity, which strongly reflects cardiovascular fitness, and showed
some improvement in muscle strength. Quality of life and sexual mood remained unaffected.

The researchers believe that bigger and longer studies in healthy elderly people would need to be
conducted before the changes observed in their study could translate in improving quality of life
and preventing or delaying frailty.
                                             # # #


                                                6
Aging
                                                                                                               Aging
P2-475
The Relationship between IGF-1 Levels and Physical Performance in Older Men and Women: The
InCHIANTI Study.
Gian Paolo Ceda*1, Fulvio Lauretani2, Cosimo Russo2, Benedetta Bartali2, Stefania Bandinelli2, Marcello Maggio1,
Anna Maria Corsi2, Domenico Valle3, Giorgio Valenti1, Luigi Ferrucci2,4. 1Dept of Intern Med and Biomed Scis,
Univ of Parma, Parma, Italy; 2Lab of Clin Epidemiology, INRCA Geriat Dept, Florence, Italy; 3Eli Lilly and Co,
Florence, Italy; 4Clin Res Br, Natl Inst of Aging, Baltimore, MD.

Aging is often associated with declining physical function, eventually leading to loss of autonomy in daily life
activities.IGF-I is a potential mediator of alterations in muscle mass and strength,and thereby of distal outcomes of
clinical importance in the elderly. However, the functional significance of the IGF-I levels in older individuals is not
completely clear. Prior observational studies have been unable to detect a relationship between IGF-I and body
composition or muscle strength after adjustment for age.Only one study has documented in a population of older
women a significant association of low IGF-I levels with poor muscle strength and mobility. Aim of this study was
to analyze the relationship between circulating IGF-1 levels and physical performance measures in a large
population of community-dwelling older subjects and to examine the existence of a gender difference.Methods: This
is part of the InCHIANTI study, a prospective population-based study of older persons, aimed to identify risk factors
for mobility-disability. The study sample consisted of 1014 participants, 447 male and 567 female, age range 20-100
years, living in the Chianti area (Tuscany). Persons with no disability in the activities of daily living (ADL) and with
a MMSE greater than 18 were included in this study. Physical performance was assessed using time to walk 400-m
and the summary performance score (SPS) of the EPESE performance battery of tests to assess lower extremity
function. IGF-1 levels were measured from frozen specimens using reagents provided by DSL. Linear regression
analyses were used to assess the relationship between log-transformed IGF-1 values and physical performance tests
after adjustment for age, BMI, knee extension torque, and level of physical activity. Analyses were performed
separately in men and women.Results: In men but not in women, IGF-1 levels were important, independent
predictor of the time to walk 400-m (p <.0001). Similar results were found with the measures of lower extremity
function. In fact,in men,IGF-1 levels were correlated with the SPS (p<0.05). Noteworthy this effect of IGF-1 on
physical performance was substantially reduced and no longer statistically significant after adjustment for knee
extension torque. In conclusion our data show a strong, sex-related, association of IGF-1 with physical performance
in older persons. This effect is likely mediated through lower extremity muscle strength.

CLINICAL POSTER: Aging (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                           7
                                                                                           Aging
P2-475 News Summary
Amino acid found to improve muscle mass in men, possibly leading to aging treatments

Insulin-like growth factor 1 (IGF-1) – an amino acid produced in response to growth hormone
(GH) – plays a major role in maintaining muscle strength and physical performance in men but
not in women, which may help in designing treatments for the decline in physical function that
accompanies aging, according to a new study being presented on Friday, June 20, at The
Endocrine Society’s 85th Annual Meeting in Philadelphia. The mechanism of this sex-related
difference is unknown, but may be related to an interaction between IGF-1 biological activity
and circulating levels of testosterone.

Geriatricians are actively searching for medicines that can help maintain or improve skeletal
muscle mass and force. During normal aging, there is gradual loss of muscle mass and
diminished capacity to reverse that loss, resulting in weakness and disability. Unfortunately,
there is no safe and effective medicine available that can slow down, arrest or reverse this
process. One of the mechanisms suggested to explain why muscles “shrink” with aging is the
progressive decline of circulating levels of hormones, such as GH, that are important for muscle
integrity and function. However, GH therapy can cause side effects and glucose intolerance.

Lately, the attention has shifted to IGF-1. Although GH exerts most of its action through IGF-1,
several studies have shown that IGF-1 is 10 times more potent than GH and positively affects
glucose metabolism and tolerance. In adults with GH deficiency, levels of IGF-1 are low.
Therapy with recombinant human GH (rhGH) stimulates the production of IGF-1, but recent
studies have shown that women need larger doses and longer treatment with GH than men.
Despite the smaller GH dose administered, the biological effect of GH treatment in terms of
body composition changes (fat free mass and total body water) is higher in men. These data
suggest that the relationship between circulating levels of IGF-1, muscle strength and physical
function is more robust in men than in women.

Dr. Gian Paolo Ceda, of the University of Parma in Parma, Italy, and colleagues in Italy and the
National Institute of Aging in Baltimore tested this hypothesis using data from InCHIANTI, a
prospective population-based study of older people aimed to identify risk factors for disability in
mobility. The study was composed of 1,014 participants, 447 men and 567 women, with ages
ranging from 20–100 years old living in the Chianti area in Italy. People who did not report
disability in activities of daily living and without severe cognitive impairment were included in
this analysis. Physical performance was assessed as time to walk 400-m and a composite score
of three tests assessing lower extremity function. Levels of IGF-1 were measured. In men but
not in women, total IGF-1 was an important, independent predictor of the time to walk 400-m
and lower extremity performance score. In addition, the effect of IGF-1 on physical performance
in men was substantially reduced and no longer statistically significant, after adjustments.

The research was supported by the Italian Ministry of Health, the U.S. National Institute on
Aging, and Eli Lilly Italia.
                                            # # #


                                                8
                                                                                                              Aging
P2-481
No Clinical Relevance of Biochemical Hypoandrogenism in Elderly Men.
Mirjam Christ-Crain*1, Beat Muller1, Thomas C Gasser1, Marius Kraenzlin1, Christian Meier1. 1Div of
Endocrinology, Dept of Intern Med, Univ Hosp, Basel, Switzerland.

Background: In males, aging is characterized by a progressive decline of testosterone, mainly non-SHBG bound
levels. The association of androgen deficiency with clinical symptoms and the ideal screening test are controversial.
Methods: We evaluated 51 healthy male volunteers between 55 and 70 years (62.2±4.6, BMI 26.6±3.2 kg/m²).
Circulating total (TT), free (FT), bioavailable testosterone (BT) and sex hormone-binding globulin (SHBG) levels
were collected after an overnight fast. Bone mineral density (BMD) was measured and questionnaires assessing
clinical symptoms, erectile function and mood (ADAM-score, IIEF-score, BDI-score) were completed.
Results: TT, FT, and BT levels (mean±SD in nmol/L) were 13.6±5.21 (range 5.9-31.0), 40.2±11.6 (22.3-70.1) and
3.7±1.8 (1.4-10.0). TT correlated moderately with FT (r=0.66). BT levels correlated with FT (r=0.54), but showed
no correlation with TT levels. Plasma SHBG levels showed a relatively weak correlation with TT (r=0.56) and BT
(r=0.43) and none with FT. Importantly, we found no correlation of TT, FT or BT values with either BMD or
clinical questionnaires. Testicular volume correlated significantly with TT levels (r=0.45), but not with FT or BT,
respectively.
Conclusion: Suspected hypogonadism is commonly verified by determining circulating hormone values. Based on
our results, testosterone values did not correlate with clinical signs such as affective symptoms, sexual interest, and
BMD. Hence, neither BT, nor TT or FT levels were reliable to mirror clinical symptoms related to age-associated
androgen deficiency. Symptoms in the aging male may therefore be rather multifactorial and unspecific and should
not be assigned to the age associated decline in testosterone levels.

CLINICAL POSTER: Aging (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                           9
                                                                                              Aging
P2-481 News Summary
Symptoms of hypogonadism are not related to testosterone deficiency, as many believe

Symptoms of hypogonadism – sexual dysfunction, breast enlargement, and muscle loss – in
aging men are not related to a testosterone deficiency, as is a common clinical assessment,
according to a new study being presented on Friday, June 20, at The Endocrine Society’s 85th
Annual Meeting in Philadelphia.

In men, aging is characterized by a progressive decline of testosterone – one of the male
hormones called androgens. The association between androgen deficiency and symptoms as
well as the ideal screening test is controversial.

To contribute to the debate, Dr. Mirjam Christ-Crain and colleagues at University Hospital in
Basel, Switzerland, investigated 51 healthy male volunteers between 55 and 70 years old.
Testosterone was measured as well as other parameters supposed to be dependent from
circulating testosterone levels. In addition, patients filled out questionnaires assessing clinical
symptoms, such as loss of libido or erectile function and mood.

Their data showed that circulating testosterone levels were not correlated with clinical signs and
symptoms of hypogonadism, such as changes in bone density, mood or lipids. Therefore,
symptoms in aging men, say the researchers, may be multifactorial and unspecific and should not
be assigned to the associated decline in testosterone levels.

The study was supported by the university hospitals of Basel.

                                               # # #




                                                 10
                                                                                                              Aging
P1-345
Lower C-Reactive Protein Concentrations with Dietary Restriction in Adult Male Rhesus Macaques.
Ricki J Colman*1,2, Joseph W Kemnitz1,2,3, Richard Weindruch1,2,4,5. 1Wisconsin Primate Res Ctr, Univ of
Wisconsin, Madison, WI; 2Inst on Aging, Univ of Wisconsin, Madison, WI; 3Dept of Physiol, Univ of Wisconsin,
Madison, WI; 4Dept of Med, Univ of Wisconsin, Madison, WI; 5Geriat Res, Educ and Clin Ctr, Veterans
Administration Hosp, Madison, WI.

Recent studies have affirmed the value of C-reactive protein (CRP), an acute-phase reactant, as a strong predictor of
negative cardiovascular events. Importantly, this relationship between CRP and coronary heart disease risk may be
independent of the presence of hyperlipidemia. Additionally, studies have shown a relationship between CRP
concentrations and insulin resistance independent of obesity. Dietary restriction (DR) is the only intervention that
has been shown to consistently extend maximum lifespan and to oppose a broad spectrum of age-associated
pathological and physiological changes in laboratory rodents. We have previously shown that adult-onset DR in
rhesus monkeys improves known risk factors for coronary heart disease, such as obesity, proteoglycan lipid binding,
lipoprotein (a) levels and insulin sensitivity. Therefore, we wished to determine if DR results in lower CRP
concentrations. We compared fasted morning serum CRP values in adult male rhesus monkeys following 12 years
of either a 30% DR (DR; n=11, mean±SEM age=21.5±0.5yrs, mean±SEM body weight=11.0±0.4kg) or ad libitum
feeding (C; n=12, mean±SEM age=21.0±0.5yrs, mean±SEM body weight=13.7±0.8kg). CRP concentrations,
measured by turbidometric assay, were lower in DR (0.50±0.0mg/dl) compared to C (0.76±0.1mg/dl) animals
(p<0.001). Additionally, CRP values correlated significantly with body weight, dual-energy x-ray absorptiometry-
measured lean body mass, fasting serum triglyceride concentration, fasting serum alkaline phosphatase
concentration and basal glucose values (C animals only). However, multiple regression analysis revealed that the
difference in CRP values between C and R groups persisted when these significant correlates were taken into
account. We conclude that long-term, moderate DR affects fasting serum CRP values above the effects of decreased
obesity. It is possible that this reduction in CRP concentration is related to decreased oxidative stress in DR animals
and can thus be added to the known benefits of dietary restriction.
          This work was supported by NIH grant PO1 AG11915.

BASIC POSTER: Aging (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                          11
                                                                                            Aging
P1-345 News Summary
Moderate changes in diet lower levels of a negative marker of heart health

Long-term, moderate dietary restriction lowers levels of C-reactive protein (CRP) – an indicator
of heart health – in rhesus monkeys, beyond the beneficial effects related to weight loss,
according to a new study being presented on Thursday, June 19, at The Endocrine Society’s 85th
Annual Meeting in Philadelphia.

Recent studies have affirmed the value of CRP as a strong predictor of negative cardiovascular
events. Moreover, this relationship between CRP and coronary heart disease risk may be
independent of the presence of hyperlipidemia. Studies have also shown a relationship between
CRP concentrations and insulin resistance, independent of obesity.

Dietary restriction is the only intervention that has been shown consistently to extend maximum
lifespan and to oppose a broad spectrum of age-associated pathological and physiological
changes in laboratory rodents. Dr. Ricki Colman and colleagues in Madison, Wis., have
previously shown that dietary restrictions in adult rhesus monkeys improves known risk factors
for coronary heart disease, such as obesity, proteoglycan lipid binding, lipoprotein (a) levels and
insulin sensitivity.

To find out more about the effect of dietary changes, Dr. Colman and colleagues studied 11 adult
male rhesus monkey after 12 years of restricting the monkeys’ diets by 30 percent and then
compared them to 12 other rhesus monkeys who ate as often as they wanted. They found lower
CRP levels in the monkeys in comparison to the other monkeys. In addition, CRP levels
correlated with body weight, lean body mass, triglycerides and basal glucose values.

This research, say researchers, lends a greater understanding of the relationship between a
marker of cardiovascular health and an aging intervention, which may further assist
endocrinologists with diagnostic interpretation. They also believe it is possible that this
reduction in CRP concentration is related to the decreased amount of oxidative stress in animals
with restricted diets and, therefore, can be added to the known benefits of dietary restriction.

This research was supported by the National Institutes of Health.

                                              # # #




                                                12
                                                                                                                  Aging
P1-348
Localization of a Gene Influencing Blood Pressure Variation on Chromosome 2q31-q34 in the Amish.
Sue Rutherford*1, Ikhide G Imumorin2, Nanette I Steine1, Braxton D Mitchell1, Alan R Shuldiner1. 1Dept of Med,
Univ of Maryland, Baltimore, MD; 2Dept of Biol, Valdosta State Univ, Valdosta, GA.

Unraveling the genetic basis of common complex disorders such as essential hypertension (EH) has been
complicated by the interaction of environmental influences, genetic heterogeneity and incomplete penetrance. Since
the etiology of hypertension begins several years before its clinical manifestation and is a major risk factor for
stroke, heart and renal failure, it is important to identify the factors that are likely to have an early influence on blood
pressure (BP) determination. Previously, we conducted a genome wide search with follow-up fine mapping for BP
variation in 691 members of the Old Order Amish population of Lancaster County, Pennsylvania, a genetically
homogeneous founder population. Peak evidence for linkage at the D2S117 marker located on chromosome 2q31-
q34 met the criteria for genome wide significance (multipoint allele sharing lodscore of 4.23, P = 0.00001) for
diastolic BP and met nominal significance (multipoint allele sharing lodscore of 1.60, P = 0.003) for systolic BP. We
now report the results of linkage disequilibrium mapping of this region with single nucleotide polymorphism (SNP)
markers. We identified a cluster of SNPs, all in strong linkage disequilibrium, showing significant association with
BP (P = 0.004 to 0.01). The associated SNPs constitute a region of less than 1 Mb at chromosome 2q32.2. The 5
positional candidate genes that reside near this region are inositol polyphosphate-1 phosphatase (INPP1),
NGF1A/EGR1 binding protein (NAB1), glutaminase phosphate activated (GLS), signal transducer and activator of
transcription-1 (STAT1) and signal transducer and activator of transcription-4 (STAT4). We conclude that a major
gene influencing BP variation resides within a 1 Mb region of chromosome 2q32.3, a region small enough for
positional cloning of the gene.

BASIC POSTER: Aging (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                            13
                                                                                             Aging
P1-348 News Summary
Amish lead researchers to possible gene responsible for high blood pressure

Amish in Lancaster, Pa., bring researchers one step closer to identifying a gene responsible for
high blood pressure, according to a new study being presented on Thursday, June 19, at The
Endocrine Society’s 85th Annual Meeting in Philadelphia.

Researchers at the University of Maryland, Division of Endocrinology, Diabetes and Nutrition
have found a region on chromosome 2 that is likely to harbor a gene that regulates blood
pressure in the Old Order Amish population of Lancaster County, Pennsylvania. This region
represents a novel susceptibility locus affecting blood pressure variation that is likely to be
relevant in other populations as well.

High blood pressure is a leading cause of death and morbidity in our society because of its
association with serious disorders such as renal failure, stroke and cardiovascular disease.
Approximately one in five individuals between the ages of 18–74 are affected by high blood
pressure and require lifelong medication, with the choice of appropriate medication currently
determined subjectively based on patient symptom and side effect responses. Individuals with
high blood pressure are likely to possess a genetic profile making them sensitive to
environmental factors, such as a high salt diet, obesity and the effects of smoking and limited
exercise. Successful identification of one or more blood pressure susceptibility genes may
reduce the occurrence of these disorders through genetic testing to identify individuals at risk for
high blood pressure so that early targeted preventive interventions may be instituted. Further
understanding of the genetics of high blood pressure may lead to the development of novel
therapeutic interventions targeted to an individual’s specific needs as well as early identification
of individuals at risk for high blood pressure for prevention of blood pressure complications.

The Amish are an excellent population for genetic studies because they keep extensive
genealogical record, allowing researchers to trace all living Amish to a small number of founding
ancestors who immigrated to the United States in the early 1700s. To identify this region on
chromosome 2, Dr. Sue Rutherford and colleagues in Baltimore and Valdosta, Ga.,
determined both systolic and diastolic blood pressures and tested more than 400 genetic markers
from 691 members of the Old Order Amish population of Lancaster County, Pennsylvania. They
compared the DNA of individuals with diastolic blood pressure above 90 mmHg with those less
than 76 mmHg and showed significant differences between these individuals in a DNA region of
less than 1 million base pairs on chromosome 2, specifically at 2q32.3.

This region is small enough to now conduct further investigations to identify a gene in this
region that may be involved in blood pressure variation.

This work was supported by research grants from the National Institute of Health and the
American Heart Association.
                                            # # #



                                                 14
Androgen Disorders
                                                          Androgen Disorders in Men & Women
P3-628
The Relationship of Testosterone to Adiposity in Ageing Men.
Carolyn A Allan*1,2, Boyd JG Strauss3, Elise A Forbes1, Henry G Burger1, Robert I McLachlan1,2. 1Prince Henry’s
Inst of Med Res, Clayton, Victoria, Australia; 2Ob and Gyn, Monash Univ, Clayton, Victoria, Australia; 3Clin Nutrit
and Metab Unit, Monash Med Ctr, Clayton, Victoria, Australia.

Increasing use of testosterone (T) replacement therapy in ageing men has drawn attention to the need to establish
normal values for serum T in this cohort. Ageing is associated with a decline in serum T levels of 1-2% per annum
with chronic illness further reducing T. Of particular importance is body composition as obese men have total T
levels 25% lower than their non-obese peers; 17% of the adult male Australian population is obese. There is little
data regarding the impact of obesity and body fat distribution on the biochemical diagnosis of hypoandrogenism in
ageing men. We are undertaking a randomized controlled trial of T replacement in otherwise healthy non-obese
(BMI<30 and abdominal circumference AC102cm) men aged55 years with symptoms consistent with androgen
deficiency. All subjects had basic anthropometry: height, weight and AC with calculation of BMI and conicity index
(CI) (0.109xACxwt/ht). AC and CI are measures of abdominal adiposity. Morning samples x2 were taken for total
T, SHBG and gonadotropins, and free T calculated (Sodergard). Recruitment allowed for screening with self-
reported BMI35 with 194 men enrolled to date. Across the cohort there were significant negative correlations
between BMI and both total T and SHBG. AC and CI were significantly related to free T in addition to total T and
SHBG. Non-obese (n=117) and obese men (n=77) showed similar mean age (63 years) and LH levels (5.5IU/L).
Obese men had lower mean total T (12.5vs.15.0nM;p<0.01), SHBG (34.9vs.40.1nM;p=0.01) and calculated free T
(269vs.297pM;p<0.05) levels than their non-obese counterparts. On multiple regression analysis AC was a more
important determinant of total and free T than BMI. The incidence of hypoandrogenism was 14% in the obese group
and 1.7% in the non-obese group according to the Endocrine Society of Australia, defined as total T<8nM or
8<T<15nM and LH>1.5xN. Our group of ageing men had an 8-fold difference in the incidence of hypoandrogenism
when stratified according to their degree of adiposity, with abdominal adiposity a more significant correlate of both
total and free T than BMI. Biochemical hypoandrogenism was very uncommon in non-obese men despite being self-
selected for symptoms ascribed to low T. These findings have important implications for the clinical management of
low T levels in obese older men.




         Supported by NHMRC, Mayne Pharma

CLINICAL POSTER: Androgen Disorders in Men & Women (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                         15
                                                 Androgen Disorders in Men & Women
P3-628 News Summary
Testosterone deficiency a problem in older obese men

Older obese men are more likely to have low levels of testosterone, which can result in numerous
other health problems, according to a new study being presented on Saturday, June 21, at The
Endocrine Society’s 85th Annual Meeting in Philadelphia.

Obesity is becoming increasingly prevalent worldwide. In the 20 years from 1980 to 2000, the
number of obese men in Australia increased from 7 percent to 17 percent. Of all adult Australian
males, 65 percent are now overweight or obese. In addition, some studies have recently
suggested a possible relationship between obesity and testosterone levels.

For these reasons, researchers in Victoria, Australia, led by Dr. Carolyn Allan, designed a cross-
sectional study to look at the relationship of testosterone levels in aging men. They recruited
men aged 55 years and older from the community through local newspaper advertisements.
Although in generally good health, the men had symptoms that have been associated with low
testosterone levels – lethargy, fatigue, depressed mood, poor short-term memory, lack of
concentration, loss of muscle strength or stamina, increased abdominal weight gain, lack of
libido and decreased sexual activity. All 223 of the participants had two morning blood tests to
measure testosterone levels, and height, weight and abdominal circumference (AC) were also
measured.

Dr. Allan and colleagues found a highly significant relationship between body composition – as
measured by BMI, weight, height and AC – and testosterone levels. Obese men had lower
testosterone levels (12.5 vs. 15.0nM) than their non-obese counterparts. Using the Australian
guidelines for diagnosis of testosterone deficiency, obese men were far more likely to be
testosterone deficient (14 percent vs. 1.7 percent). Non-obese men were very unlikely to be
testosterone deficient despite the fact that only men with testosterone-deficient–like symptoms
were included in the study.

As a result of the greater numbers of obese men, say the researchers, it is likely that the number
of symptomatic men with low testosterone levels will dramatically increase in the future, having
important implications for the clinical management of this group of men.

This work forms the basis for an ongoing randomized placebo-controlled study of the effects of
testosterone replacement on aging men.

This research is supported by a scholarship from the National Health and Medical Research
Council and the National Heart Foundation and Mayne Pharma, all located in Australia.

                                              # # #




                                                16
                                                            Androgen Disorders in Men & Women
P3-598
Polycystic Ovary Syndrome: Effects on Quality of Life and Sexuality.
Susanne Hahn*1, Sigrid Elsenbruch2, Daniela Kowalsky2, Manfred Schedlowski2, Klaus Mann1, Onno E Janssen1.
1
  Dept of Med, Div of Endocrinology, Univ of Essen, Essen, NRW, Germany; 2Dept of Med Psych, Univ of Essen,
Essen, NRW, Germany.

Polycystic Ovary Syndrome (PCOS) is a common endocrine disorder characterized by chronic anovulation in
combination with hyperandrogenism, affecting approximately 5% of premenopausal women. PCOS is one of the
leading causes of infertility and exhibits a variety of clinical signs, like hirsutism, acne, seborrhoea, hair loss, and
obesity. These clinical manifestations of PCOS are thought to cause the profound psychological distress found in
affected women. In addition, disturbances in sexual attitude and behavior may be expected in PCOS women. To
investigate the impact of PCOS on health-related quality of life (HRQL) and psychosocial well-being 50 PCOS
women (age 18-35) and 50 control women (age 18-37) were compared using three standardized validated
questionnaires, the German version of the SF-36, the “Fragebogen zur Lebenszufriedenheit “(FLZ) and the German
version of the SCL-90-R. On a five-point rating scale (“not at all “ to “very much”) women rated the impact of
hirsutism, obesity and infertility on their life. Sexual attitude and behavior were assessed using a 100-mm visual
analogue scale (VAS). PCOS patients and healthy controls did not differ in sociodemographic variables, including
education, family status and employment. PCOS women showed greater psychological disturbances, especially on
the SCL-90-R dimensions: Obsessive-Compulsive, Interpersonal Sensitivity, Depression, Anxiety and Aggression
(p < 0.05). In PCOS-women, analysis of the FLZ reported a lower degree of satisfaction in the areas: Health, Self
and Sex compared with the controls (p < 0.01). HRQL measured with the SF-36-questionnaire showed significantly
decreased scores for Physical Role Function, Bodily Pain, Vitality, Social Function, Emotional Role Function and
Mental Health in PCOS-patients (p < 0.05). For the scales Physical Function and General Health no differences
were found between the groups. PCOS-women and controls were comparable regarding partner status and frequency
of sexual intercourse, however PCOS affected patients were significantly less satisfied with their sex life (p < 0.001)
and found themselves less sexually attractive (p < 0.001). In conclusion, PCOS severely affects health-related
quality of life and sexuality.

CLINICAL POSTER: Androgen Disorders in Men & Women (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                          17
                                                  Androgen Disorders in Men & Women
P3-598 News Summary
Endocrine problem in women reduces quality of life and sexual satisfaction

A common endocrine problem in women called polycystic ovary syndrome (PCOS) causes a
major reduction in quality of life and severely limits sexual satisfaction in affected women,
according to a new study being presented on Saturday, June 21, at The Endocrine Society’s 85th
Annual Meeting in Philadelphia. In addition to medical treatment, psychological counseling and
participation in PCOS self-help groups should be offered, recommend researchers.

PCOS is the most common hormonal disturbance of young adult women. PCOS is one of the
leading causes of infertility and manifests with irregular or absent periods, excess facial or body
hair, acne, hair loss and obesity, leading to psychological distress and negative effects on self-
esteem. Women with PCOS also have an increased risk of diabetes, heart disease and
endometrial cancer in later years.

For the first time, researchers at the University of Essen in Essen, Germany, led by Dr. Onno E.
Janssen, evaluated quality of life, life satisfaction and sexuality in 50 adult PCOS patients and 50
control women with a comprehensive combination of standardized questionnaires and a
collection of PCOS-specific questions. PCOS patients and healthy controls did not differ in
sociodemographic variables, including education, family status and employment. However,
PCOS women were affected by psychological disturbances resulting in reduced quality of life
and psychosocial well-being. PCOS women and controls were comparable regarding partner
status and frequency of sexual intercourse, but PCOS patients were significantly less satisfied
with their sex life, found themselves less sexually attractive and believed that their outer
appearance makes it difficult to form social contacts.

In PCOS patients, the researchers advise that both medical and psychological aspects should be
taken into account during treatment.

                                               # # #




                                                18
                                                          Androgen Disorders in Men & Women
P3-629
The Prevalence of Hypogonadism in Men with Coronary Artery Disease: Diabetic Subgroup Analysis.
Christopher J Malkin*1, Peter J Pugh1, Richard D Jones2, Paul D Morris1, Sonia Asif1, Kevin S Channer1, Hugh T
Jones2. 1Dept of Cardiol Res, Royal Hallamshire Hosp, Sheffield, South Yorkshire, United Kingdom; 2Acad Unit of
Endocrinology. Div of Genomic Med, Univ of Sheffield, Sheffield, South Yorkshire, United Kingdom.

Background: Low serum testosterone levels are associated with risk factors for coronary artery disease (CAD)
including diabetes and obesity. Men with CAD have lower androgen levels than matched controls (1). Testosterone
treatment reduces symptoms and ischaemia in men with angina (2). This is a population of men prone to testosterone
deficiency that may especially benefit from testosterone replacement. In this study we have determined the size of
the problem and compared testosterone levels of both men with and without diabetes and impaired glucose tolerance
(IGT).
Methods Men were recruited on day of angiography and blood samples were taken before 0930 hours. 871/1146
men had CAD with >70% stenosis in an epicardial coronary artery. 69 subjects with evidence of inflammation were
excluded. Serum levels of total (TT) and bio-available testosterone (BT) (non-SHBG-bound T) were measured in
the remaining 802. Hypogonadism was defined as TT<7.5 nmol/L and / or BT <2.5 nmol/L. Subjects were divided
into 3 groups. (1) Diabetic: known diabetes or random blood sugar (RBG) >11.1 mmol/l. (2) IGT: RBG between 7
mmol/l and 11.1 mmol/l. (3) Normal: RBG <7mmol/l.
Results. The overall prevalence of hypogonadism was 23.4%. Sub-group analysis: prevalence in group (1)
29.6%(n= 135), (2) 34.7%(n=46) and (3) 21.3%(n=621). The prevalence in the pooled diabetic/impaired glucose
tolerance group was 30.9% which was significantly higher than the group with normal RBG (2 =12,p=0.005). The
pooled group (diabetes/IGT) had a higher BMI (29.5±03 v 27.7±0.2, p=0.0001), drank less alcohol (7.4±0.7 v
9.3±0.5, p=0.028), had lower TT (12.7±0.5 v 14.2±0.3, p=0.007), and lower SHBG (38.3±0.2 v 45±0.1, p=0.005).
Conclusions. Using strict criteria for the diagnosis of hypogonadism nearly a quarter of men with CAD are
biochemical testosterone deficient. The prevalence is higher in men with diabetes or impaired glucose tolerance. The
lower total testosterone may be explained by increased obesity. Whether or not these would benefit from
testosterone replacement would need further investigation with a large outcome study.
          Reference: 1. English et al. Men with coronary artery disease have lower levels of androgens than men with
normal coronary angiograms. Eur Heart J 2000;21:890-4.
          2. English et al. Low dose transdermal testosterone therapy improves angina threshold in men with stable
angina; a randomised, double blind, placebo controlled study. Circulation 2000;102:1906-1911.

CLINICAL POSTER: Androgen Disorders in Men & Women (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                        19
                                                 Androgen Disorders in Men & Women
P3-629 News Summary
Large proportion of men with heart disease have testosterone deficiency;
higher in diabetics

Nearly one-quarter of men with heart disease have low levels of testosterone, which has been
linked to heart and other health problems, according to a new study being presented on Saturday,
June 21, at The Endocrine Society’s 85th Annual Meeting in Philadelphia. The risk of a
testosterone deficiency was higher in diabetic, obese and older men.

Men suffer more than twice the incidence of coronary heart disease than women, which is not
due to differences cardiovascular risk factors, such as smoking and cholesterol. It is widely
believed that testosterone is bad for the heart and responsible in some way for the excess disease
seen in males. However, current evidence contradicts this notion. First, if men with angina are
treated with even relatively small doses of testosterone, they develop less chest pain and feel
better. Second, it seems that men with coronary heart disease – as diagnosed by a coronary
angiogram – have testosterone levels lower than that of men with normal heart X-rays. Finally,
low levels of testosterone are linked with a number of cardiovascular risk factors, such as
adverse cholesterol profiles, insulin resistance and adverse clotting profiles.

Dr. Hugh Jones and colleagues in Sheffield, United Kingdom, sought to determine the size of the
problem of testosterone deficiency in men with heart disease. They compared testosterone levels
of 802 men both with and without diabetes and impaired glucose tolerance. Men with low
testosterone are called hypogonadal; they may have vague symptoms, such as depresion and
mood swings, reduced physical strength or endurance, erectile dysfunction and long-term risk of
osteoporosis.

More than 23 percent of men were hypogonadal. The patients with diabetes or high fasting sugar
levels, who exhibited an even higher risk of hypogonadism, were more obese, which may
explain the lower testosterone levels because fat cells convert testosterone to estrogen. In
addition, patients with diabetes are recognized to have more aggressive and severe heart disease,
say the researchers, although they were unable to confirm this aspect in their study.

                                              # # #




                                                20
                                                            Androgen Disorders in Men & Women
P3-632
Prevalence of Polycystic Ovaries in Patients with the Polycystic Ovary Syndrome (PCOS) or with 21-
Hydroxylase (21-OH) Deficient Non-Classic Adrenal Hyperplasia (NCAH).
Duarte Pignatelli*1, Jorge Beires2, Ricardo Azziz3. 1Endocrinologia, Fac de Med do Porto, Porto, Portugal;
2
  Ginecologia, Fac de Med do Porto, Porto, Portugal; 3Ob/Gyn, Cedars-Sinai Med Ctr, Los Angeles, CA.

The presence of polycystic ovaries on transvaginal ultrasound (TV-U/S) has been used to signal the presence of the
endocrine disorder PCOS. However, we have previously suggested that this sonographic appearance was a non-
specific marker of hyperandrogenic anovulation, as a significant proportion of patients with 21-OH deficient NCAH
also demonstrate “polycystic” ovaries on TV-U/S. To determine the value of ovarian TV/US in the diagnosis of
PCOS, we studied the prevalence of “polycystic” ovaries in consecutive PCOS and NCAH patients. MATERIALS
AND METHODS: We prospectively determined the ovarian morphology sonographically in 22 consecutive women
with PCOS and 16 patients with NCAH. PCOS was diagnosed by the presence of oligo-ovulation (i.e. menstrual
cycles 8 cycles/yr) in the presence of hirsutism (modified Ferriman-Gallwey score 8) and hyperandrogenemia
(total testosterone 86.0 ng/dL, free testosterone 3.0 pg/mL, androstenedione 3.2 ng/mL, and/or DHS 4300
ng/mL), after the exclusion of 21-OH deficient NCAH, thyroid dysfunction and hyperprolactinemia. These criteria
are consistent with those proposed by an NIH-sponsored meeting in 1990. 21-OH deficient NCAH was diagnosed
by a 17-hydroxyprogesterone level 60 minutes after acute IV ACTH stimulation 10 ng/mL. Ultrasonography of the
ovaries was performed using a 7MHz transvaginal transducer (Aloka 5000, Japan), and the procedure was
performed between days 3 and 8 in the majority of patients. Polycystic ovaries were defined as an increased mean
ovarian volume (> 11 mL) and/or the presence of 12 mean follicles/ovary measuring 2 mm to 9 mm. RESULTS:
Overall, 6 of the 22 (27.3%) PCOS patients demonstrated “polycystic” ovaries on TV-U/S, compared to 3 of 16
(18.7%) NCAH women, a difference not reaching statistical significance. The presence of “polycystic” ovaries was
not correlated with any of the clinical features, including age, BMI, hirsutism score, androgen levels, or basal insulin
or glucose levels. CONCLUSIONS: In this prospective study approximately 25% of PCOS patients diagnosed
endocrinologically had “polycystic” ovaries on TV-U/S, a prevalence similar to that of 21-OH deficient NCAH.
These data suggest that “polycystic ovaries” may reflect hyperandrogenic ovulatory dysfunction, and that ovarian
ultrasonography is an insensitive and non-specific method of diagnosing PCOS.

CLINICAL POSTER: Androgen Disorders in Men & Women (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                          21
                                                  Androgen Disorders in Men & Women
P3-632 News Summary
Ultrasound not a reliable in diagnosing specific syndromes marked by ovarian cysts

Ultrasound appearance of polycystic ovaries is not an accurate way to distinguish the various
syndromes marked by excess body and facial hair growth in women, according to a new study
being presented on Saturday, June 21, at The Endocrine Society’s 85th Annual Meeting in
Philadelphia.

The frequency of hirsutism in women – the excess growth of body or facial hair – is very high
and this disease greatly affects quality of life. In addition to the aesthetic problem, these women
often have irregular periods and infertility. There is a need to distinguish those cases that have
polycystic ovary syndrome (PCOS) – a common endocrine problem in women – because they
represent a more complex form of disease, precisely the cases with irregular menses and
infertility. It is also known that women with PCOS have insulin resistance and, therefore, are
more prone to develop diabetes and cardiovascular disease.

The presence of polycystic ovaries on transvaginal ultrasound (TV-U/S) has been used to signal
the presence of the PCOS. However, researchers have previously suggested that this
sonographic appearance was a non-specific marker of hyperandrogenic anovulation, as a
significant proportion of patients with non-classic adrenal hyperplasia (NCAH) also demonstrate
“polycystic” ovaries on TV-U/S. NCAH is an endocrine-related disorder that manifests as
hirsutism, irregular periods, among other possible symptoms. To determine the value of ovarian
TV/US in the diagnosis of PCOS, Dr. Duarte Pignatelli and colleagues in Portugal and Los
Angeles studied the prevalence of “polycystic” ovaries in consecutive PCOS and NCAH
patients.

In this prospective study, approximately 25 percent of PCOS patients diagnosed
endocrinologically had “polycystic” ovaries on TV-U/S, a prevalence similar to that of patients
with NCAH. These data suggest that “polycystic ovaries” may reflect hyperandrogenic
ovulatory dysfunction, and that ovarian ultrasonography is an insensitive and non-specific
method of diagnosing PCOS.

This study was supported by the Portuguese National Health System.

                                              # # #




                                                22
                                                           Androgen Disorders in Men & Women
P3-593
Premature Pubarche (PP): Predictors of Risk for Persistent Hyperandrogenism (HA).
Selma F Witchel*1, Christopher E Aston2. 1Ped Endocrinology, CHP/Univ of Pittsburgh, Pittsburgh, PA; 2Genetic
Epidemiology Unit, Oklahoma Med Res Fdn, Oklahoma City, OK.

PP may be an early manifestation of persistent HA  incipient polycystic ovary syndrome (PCOS). Women with
PCOS have an increased risk to develop IGT, T2DM, coronary artery disease,  endometrial cancer. Familial
clustering of PCOS implies a role for genetic influences. However, while PP may precede PCOS, not all girls with
PP develop PCOS. To identify risk factors that predict progression from PP to PCOS, we have prospectively
followed 30 girls initially referred for PP. Methods: Blood samples were obtained for hormone determinations 
genetic analyses (CYP21  HSD3B2 genes  for variants of the IRS-1, ADRB, PPAR-2,  UGT2B15
genes).Results: Mean follow-up was 8.6±3.1 years with mean current age being 16.4±3.3 years; 13 had regular
menstrual cycles, 14 had irregular menses (cycles > 35 days), 2 were in their first gynecologic year,  1 had not yet
experienced menarche. The girls were classified into persistent (P) (elevated androgen concentrations at last follow-
up, n=21)  regressant (R) (normal androgen concentrations at last follow-up, n=9). Mean androstenedione (AND)
 free testosterone concentrations were significantly greater (p<0.0001 and p<0.05, respectively) and mean SHBG
significantly lower (p<0.005) among the P than the R group. Mean Ferriman-Gallwey score  mean BMI were
significantly greater among the P group (p<0.05). At the initial evaluation, mean chronologic age, mean bone age,
mean breast development, mean pubic hair development, mean AND,  ACTH-stimulated responses did not differ.
Mean basal 17-OHP  cortisol concentrations were significantly greater among the P group (p<0.05). Mean SHBG
concentrations were significantly lower (p<0.01) among P than R group at presentation. At timepoints 1  2, mean
SHBG concentrations were 0.85±0.54  0.58±0.39 for the P group,  1.55±0.43  1.44±0.49 for the R group. The
IRS-1 PPAR-2 variants occurred exclusively in the P group. One HSD3B2 variant carrier  4/5 CYP21 mutation
carriers progressed to persistent HA. Conclusions: Advanced skeletal age, higher basal 17-OHP concentrations, 
lower SHBG concentrations at the initial evaluation  greater weight gain during the peri-pubertal years were
associated with an increased risk to develop persistent HA. Heterozygosity for the G972R variant of the IRS-1 gene
and/or the P12A variant of the PPAR-2 gene may be genetic markers indicative of propensity to develop persistent
HA and incipient PCOS (Odds ratio = 17.2, p=0.026).
          Supported by NIH & AHA

CLINICAL POSTER: Androgen Disorders in Men & Women (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                         23
                                                 Androgen Disorders in Men & Women
P3-593 News Summary
Adolescent weight gain and genetic markers identify early maturing girls with likelihood of
endocrine problem

Girls who develop pubic hair at an early age are more likely to develop a common endocrine
problem if they show an increase in weight gain during early adolescence and have several
genetic markers, according to a new study being presented on Saturday, June 21, at The
Endocrine Society’s 85th Annual Meeting in Philadelphia.

Some girls with early development of pubic hair develop polycystic ovary syndrome (PCOS)
when they are adolescent or young adults. Women with polycystic ovary syndrome have an
increased risk of developing impaired glucose tolerance, diabetes and endometrial cancer.
Lifestyle interventions to prevent obesity have been reported to decrease the rate at which adults
with impaired glucose tolerance develop diabetes.

The goal of the current study was to characterize clinical features, hormone levels and genetic
markers that predict which girls with early development of pubic hair will develop PCOS so that
these girls and their families can be targeted for early intervention.

For this study, Drs. Selma Feldman Witchel of the University of Pittsburgh and Christopher E.
Aston of the Oklahoma Medical Research Foundation in Oklahoma City followed a group of 30
girls from the time of presentation with early development of pubic hair until mid-adolescence.
They found that increased weight gain during early adolescence and several genetic markers
appeared to be more common among girls who developed PCOS compared to those who did not.

This research was supported by the National Institutes of Health and the American Heart
Association.

                                              # # #




                                                24
                                                             Androgen Disorders in Men & Women
P3-622
Prostate Volume and Growth in Testosterone-Substituted Hypogonadal Men Are Dependent on the CAG
Repeat Polymorphism of the Androgen Receptor Gene: A Longitudinal Pharmacogenetic Study.
Michael Zitzmann*1, Marion Depenbusch1, Joerg Gromoll1, Eberhard Nieschlag1. 1Univ Clin, Inst of Reprod Med,
Muenster, NRW, Germany.

Testosterone (T) substitution in hypogonadal men results in growth of the prostate gland. T effects are mediated via
the androgen receptor (AR). The length of the (CAG)n polymorphism of the AR gene is negatively associated with
transcriptional activity and might account for variations in prostate growth during substitution therapy. In 131
hypogonadal men aged 18 to 69 years, we assessed prostate volume longitudinally by transrectal ultrasonography
and determined AR (CAG)n, sex hormone levels and anthropometric measures. 69 men with primary and 62 with
secondary hypogonadism began substitution therapy with intramuscular injections of T enanthate (n = 81),
transdermal T preparations (n = 19), subcutaneous injections of human chorionic gonadotropin (n = 17) or oral T
undecanoate (n = 14) for 2.4 ± 0.8 years. Average prostate size increased from 15.8 ± 6.1 ml to 23.0 ± 6.8 ml.
Analysis of variance including covariates revealed initial prostate size to be dependent on age (p < 0.001) and
baseline T levels (p = 0.01), but not on number of (CAG)n (ranging from 13 to 30, mean 21.4 ± 3.5). Prostate
growth per year and absolute prostate size under substituted T levels (baseline levels: 6.1 ± 3.3, substitution levels:
21.6 ±10.3 nmol/L) were strongly dependent on (CAG)n, with lower treatment effects in longer repeats (both p <
0.001). Other significant predictors were initial prostate size (negative for growth rate and positive for absolute size)
and age (positive for both growth rate and absolute size). The odds ratio for men with (CAG)n < 20 compared to
those with (CAG)n  20 to develop a prostate size of at least 30 ml under T substitution was 8.7 (95% CI 3.1 – 24.3,
p < 0.001). This observation was strongly age-dependent with a more pronounced odds ratio in men older than 40
years. This first pharmacogenetic study on androgen substitution in hypogonadal men demonstrates a marked
influence of the AR gene (CAG)n polymorphism on prostate growth.

CLINICAL POSTER: Androgen Disorders in Men & Women (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                           25
                                                  Androgen Disorders in Men & Women
P3-622 News Summary
Genetic factor identifies men on testosterone therapy who may develop enlarged prostate

In men who need to have testosterone therapy, a genetic factor can identify those at a higher risk
of developing an enlarged prostate gland before the start of therapy and be closely monitored or
given an adjusted testosterone dose, according to a new study being presented on Saturday, June
21, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

With advancing age or due to specific diseases, many men lack sufficient levels of testosterone,
which can result in a decrease of psychological factors, such as vigor, cognition and libido. It
can also negatively affect physical traits, such as bone density, red blood cell count and muscle
mass. Testosterone-substitution therapy is a useful tool to help these men. The prostate as a
testosterone-sensitive organ will grow under such a therapy, but growth and total organ size
show a great variety among patients. Some men exhibit a pronounced growth of the prostate
when administered testosterone, which can cause health problems; some men do not.

Dr. Michael Zitzmann and colleagues at the Institute of Reproductive Medicine in Munich
initiated testosterone therapy in 131 men to find out whether a specific, genetically determined
factor has any influence on prostate growth. This factor is the CAG repeat chain in the androgen
receptor gene, which ranges from 10 to 37 repeats in normal men. The androgen receptor
facilitates testosterone action, and the longer the CAG repeat chain is, the less the receptor will
activate the processes initiated by testosterone, a finding previously shown in cell cultures.

In the current study, the men with shorter CAG repeat chains exhibited a much more pronounced
growth of the prostate than those with longer chains because testosterone works more efficiently
in them. In men with less than 20 repeats and especially in men older than 40, there was an
eight-fold higher risk to be diagnosed with a prostate larger than 30ml – a size in which
symptoms of voiding problems usually start – than the other men.

The study could not investigate the risk to develop prostate cancer, as it was not observed in the
patients.

This study was supported by the German Research Society.

                                              # # #




                                                26
Cardiovascular Endocrinology
                                                                         Insulin-like Growth Factors
OR35-4
Hsp60 Amplified IGF-1 Receptor Signaling in Cardiac Muscle: Functional Implication on Declined
Myocardial Protection in Diabetic Cardiomyopathy.
Yue-xin Shan*1, Tung Lin Yang1, Ahmad Samsamshariat1, Ruben Mestril2, Ping H Wang1. 1Med, Univ of
California, Irvine, CA; 2Physiol, Loyola Univ, Chicago, IL.

Declined myocardial protection is a major feature of diabetic cardiomyopathy. Using DNA microarray, we
discovered that the expression of Hsp60 was reduced in the myocardium of STZ-DM rats. Hsp60 is a newly
recognized cardiac protective heat shock protein, but the role of Hsp60 in diabetic cardiomyopathy has never been
investigated. Western blots showed that the abundance of Hsp60 protein was reduced by 52% in diabetic
myocardium, accompanied by 40% reduction in IGF-1 receptor  subunits (IGF1R). Further study indicates that
insulin deficiency, not hyperglycemia, was the cause of Hsp60 reduction in diabetic myocardium. To define whether
decreased IGF1R was related to reduction of Hsp60, primary cardiomyocytes were transduced with Hsp60 via
adenoviral vector. Compared to the cells transduced with control vector, overexpression of Hsp60 increased the
abundance of IGF1R by 2-fold. IGF-1-stimulated IGF1R autophosphorylation was also increased by 2.8-fold in the
cardiomyocytes overexpressing Hsp60. Moreover, the stoichiometry of IGF1R phosphorylation/receptor was
enhanced by Hsp60 overexpression, suggesting that Hsp60 overexpression increased the number of functioning
receptors and thus amplified activation of IGF1R signaling. In the cardiomyocytes overexpressing Hsp60, IGF-1
activation of Mek, Erk, p90Rsk, and Akt were accordingly amplified and the anti-apoptosis action of IGF-1 was
enhanced. The effects of Hsp60 overexpression on IGF-1 signaling were observed at physiological and
pharmacological concentrations of IGF-1. Transducing cardiomyocytes with anti-sense Hsp60 oligo reduced Hsp60
expression by  40%, decreased the abundance of IGF1R by 45%, attenuated IGF1R autophosphorylation,
decreased Erk activation, and attenuated the anti-apoptosis action of IGF-1. Scambled control oligo did not affect
IGFIR signaling. Using cycloheximide to inhibit protein synthesis did not alter the effect of Hsp60 on IGF1R
signaling, suggesting that Hsp60 modulates IGF1R signaling through protein chaperoning. IGF-1 signaling enhances
myocardial survival, and we have recently shown that Hsp60 protects cardiomyocytes against apoptotic injury.
Conclusions: 1] Decreased Hsp60 expression and subsequent decline of IGF1R signaling may be a fundamental
mechanism underlying the development of diabetic cardiomyopathy, 2] Hsp60 represents a novel regulatory
mechanism that modulates IGF1R signaling.

BASIC ORAL: Insulin-like Growth Factors (11:15 AM - 12:45 PM)

Presentation Date: 6/22/2003, Time: 12:00 PM; Location: 108A/B




                                                       27
                                                                 Insulin-like Growth Factors
OR35-4 News Summary
Gene therapy shows promise for protecting heart muscle from injury in diabetics

Gene therapy with two proteins restored the levels of one of the proteins in the heart muscle of
diabetic animals, making the heart muscle more resistant to injury, according to a new study
being presented on Sunday, June 22, at The Endocrine Society’s 85th Annual Meeting in
Philadelphia.

Heart disease is the leading cause of death among diabetic patients. Diabetic patients often have
weakened heart muscle that may lead to the development of heart failure. The same weakened
heart muscle also increases the risk of death and complications in the event of a heart attack.

Dr. Ping H. Wang and colleagues in Irvine, Calif., and Chicago studied whether two proteins,
Hsp10 and Hsp60, can regulate a cell surface receptor – insulin-like growth factor 1 (IGF-1)
receptor – that commands the survival signals in heart muscle. The results showed that Hsp10
and Hsp60 could increase the survival signals sent out by the IGF-1 receptor in heart muscle.
Hsp10 and Hsp60 did so by preventing the IGF-1 receptor from destruction.

In two animal models of diabetes, the researchers previously observed that the level of Hsp60
was significantly reduced in the diabetic cardiac muscle, which is accompanied by a reduction of
IGF-1 receptor due to increased destruction of IGF-1 receptor. When they genetically
manipulated Hsp60 and reduced the level of Hsp60 in cardiac muscle cells, IGF-1 receptor
survival signals were reduced and cardiac muscle cells became less viable. These results suggest
that Hsp10 and Hsp60 may play a fundamental role in the regulation of cardiac muscle survival
in diabetes.

These experiments also showed that transferring the Hsp60 gene to cardiac muscle cells
enhanced the survival signaling commanded by IGF-1 receptor, thereby raising the possibility
that gene therapy with Hsp10 and Hsp60 may be used to restore the level of Hsp60 in the heart
muscle of diabetic patients and make the heart muscle more resistant to injury.

This study was supported by the National Institutes of Health.

                                              # # #




                                                28
                                                                                  Signal Transduction II
P2-51
Ucn-II Is Expressed in Cardiac Myocytes and Is Cardioprotective Against Ischemia Reperfusion Injury Via
Activation of a MAPK Pathway: An Essential Endogenous Cardioprotective Role for CRFR2 in the Murine
Heart.
Bhawanjit K Brar*1, Anne K Jonassen2, Elena M Egorina2, Alon Chen1, Marilyn H Perrin1, Ole D Mjos2, Wylie
Vale1. 1Clayton Fdn Labs for Peptide Biol, Salk Inst, La Jolla, CA; 2Med Physiol, Inst of Med Biol, Fac of Med,
Univ of Tromso, Tromso, Norway.

Corticotropin-Releasing factor (CRF) and the related peptide, Urocortin (Ucn-I) have a variety of effects on the
cardiovascular system. Ucn-I is cardioprotective against ischemic reperfusion injury (I/R). Recently, two new
members of the CRF/Ucn-I gene family have been discovered, Urocortin-II/Stresscopin Related Peptide (Ucn-II)
and Urocortin-III/Stresscopin (Ucn-III). Ucn-II and Ucn-III are highly selective for CRF type 2 beta (CRFR2b)
which mediates the cardiovascular actions of Ucn-I. Ucn-I activates both CRFR2b and CRF receptor type 1, the
receptor that mediates the endocrine stress response. Therefore, Ucn-II and Ucn-III would be preferred ligands to
treat I/R injury. We examined the relative cardioprotective effects of Ucn-II, Ucn-III, Ucn-I and CRF in wild type
mouse cardiac myocytes and investigated the signaling mechanisms of protection. Ucn-II, but not Ucn-III is
expressed in cardiac myocytes suggesting that endogenous Ucn-II may have a physiological role in the heart. Ucn-
II, Ucn-III, CRF and Ucn-I activate CRFR2 in cardiac myocytes resulting in ERK1/2 phosphorylation which is
regulated by the Ras/Raf-1 kinase pathway and is independent of adenylate cyclase and therefore of cAMP
activation. Cardiac myocytes isolated from CRFR2 null mice are less resistant than wild type cells to simulated I/R
injury suggesting that CRFR2 may be essential for preventing ischemic induced cell death. Ucn-I, Ucn-II, Ucn-III
protect cardiac myocytes from ischemic induced cell death when the peptides are added to the cells before and after
ischemia and the mechanism of protection is via activation of ERK1/2. In contrast to the urocortins, CRF protects
cardiac myocytes from simulated hypoxic/ischemic injury when administered before ischemia only and the
protective effect of CRF is independent of ERK1/2. Using the Langendorff perfused rat heart system we show that
the mechanism of cardioprotection by Ucn-II is via activation of ERK1/2 and confirm that the CRF pre-ischemic
cardioprotective effect is independent of ERK1/2. We propose the use of the CRFR2 selective peptides, Ucn-II or
Ucn-III to treat ischemic heart disease because they are less likely than Ucn-I or CRF to activate the hypothalamic
pituitary axis at pharmacological doses.
This work was supported by National Institute of Health Grants DK-26741 (W.V.), The Robert J. and Helen C.
Kleberg Foundation, Foundation for Research (W.V., M. P.), British Heart Foundation Fellowship (FS- 200173) (B.
K. B.)

BASIC POSTER: Signal Transduction II (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                        29
                                                                       Signal Transduction II
P2-51 News Summary
Newly found hormones may hold keys to protect against heart damage

Newly discovered hormones found in the heart and the brain may be potential therapies for
reducing damage to the heart caused by inadequate blood flow, according to a study of rodents
being presented on Friday, June 20, at The Endocrine Society’s 85th Annual Meeting in
Philadelphia.

When blood containing oxygen and nutrients cannot be delivered to the heart, a process known
as ischemia, the heart muscle cells die. Ischemia results in the heart becoming weaker, which
leads to heart failure and death. Ischemic heart disease effects millions of people in the United
States and worldwide.

Urocortin is a hormone that is found in the brain as well as in heart muscle cells. When rat and
mouse heart cells are treated with urocortin both before and after simulated ischemia, the heart
muscles are able to survive. Urocortin acts through two receptors known as type 1 and type 2.
The type-1 receptor is found in the pituitary region of the brain and elsewhere and is involved in
many stress-related actions. The second receptor, type 2, is found in the heart. Unfortunately,
while benefiting the heart, urocortin also exerts stress-like actions on the pituitary gland.

Recently, two new hormones have been discovered that are similar to urocortin, known as
urocortin 2 and urocortin 3. Urocortin 2 and 3 act on the receptor in the heart (type 2) and not
the pituitary (type 1); therefore, they may be superior to urocortin in preventing heart cells from
dying from ischemia.

Therefore, Dr. Bhawanjit K. Brar and colleagues from the University of Tromso in Tromso,
Norway, investigated whether urocortin 2 or urocortin 3 could protect mouse heart cells in
culture from ischemia as well as rat hearts from regional ischemia, when the coronary artery is
blocked.

Their results show that when mouse heart muscle cells or the rat heart is treated with urocortin 2
or urocortin 3 before or after ischemia both hormones protect against muscle cell death. The
data also indicate that urocortin 2 is found in heart muscle cells, revealing a potential natural
mechanism by which the heart can protect itself from ischemia. Dr. Brar and colleagues also
show that the mechanism by which urocortin 2 and urocortin 3 are protective against ischemia is
by activating an enzyme known as mitogen activated protein kinase. In addition, removal of the
urocortin 2 and 3 receptor in heart muscle cells increases the percentage of these cells that die
from ischemia compared to normal heart muscle cells that have this receptor, highlighting the
possible significance of the urocortin system in the heart.

This research was supported by a British Heart Foundation International Fellowship, the
National Institutes of Health, The Robert J and Helen C. Kleberg Foundation and in the
Foundation for Research.
                                              # # #


                                                 30
                                                                                   Signal Transduction II
P2-58
Etanercept Protection from TNF-Induced Reduction of -Adrenergic Receptor Kinase, Rapid Ventricular
Tachyarrhythmias and Infarct Size Reduction in Canine Models of Myocardial Infarction.
Xichun Yu*1, Eugene Patterson1,2, David C Kem1. 1Med, Univ of Oklahoma Hlth Scis Ctr, Oklahoma City, OK;
2
  Cell Biol, Univ of Oklahoma Hlth Scis Ctr, Oklahoma City, OK.

TNF increases after myocardial ischemia and may modify infarct size, signal transduction and the ultimate
development of lethal ventricular tachyarrhythmias (VT). To determine if anti-TNF treatment will alter infarct
size, dogs were injected with 2 mg/kg hTNFR:Fc (etanercept) or saline 24-hr and 1-hr prior to 2-hr occlusion/6-hr
reperfusion of the left anterior descending (LAD) artery. Infarct size was decreased when expressed as % area at risk
(47%) or left ventricular mass (51%) in the etanercept group (n = 8) compared to saline (n = 9, p < 0.05). Leukocyte
infiltration (myeloperoxidase activity) was markedly decreased in the etanercept group compared to saline controls
(n = 7, p < 0.01). We previously reported that ischemic subepicardial border zone (EBZ) tissue has markedly
decreased -adrenergic receptor kinase (ARK) activity leading to abnormal desensitization to isoproterenol 6-24 hr
after coronary artery ligation (CAL) in the dog (1). We have pretreated 16 dogs with 2 mg/kg etanercept (n = 7) or
saline (n = 9) 24-hr and 1-hr prior to CAL and monitored for 24-hr by ECG. The EBZ and non-ischemic remote site
(RS) tissues were collected at 24-hr. ARK protein was measured by quantitative immunoblot. Expressed as
percentage of unoperated control tissue, the EBZ in the saline group had a marked drop of ARK to 4.3 ± 2.2 %.
The etanercept group dropped only to 33.8 ± 7.2 % (p < 0.01, n = 5), representing an 8-fold salvage of ARK
expression in EBZ tissue. There was no significant difference of ARK content between the RS in saline and
etanercept groups (87.4 ± 8.2 vs 103 ± 16.2, p = 0.41, n = 5). A decreased frequency of rapid (360 bpm) ventricular
triplets capable of initiating rapid monomorphic VT and sudden cardiac death (SCD) was observed in the etanercept
group (6 ± 6/hr) as compared to the control group (49 ± 40/hr, n = 7, p < 0.05) from 3 - 24 hrs. 2 of 9 control
animals died from SCD while none of the etanercept-treated animals died. These data support a role for TNF on
infarct size and ARK expression. TNF antagonism may attenuate SCD following CAL by partially protecting
ARK-mediated desensitization of -adrenergic receptors in the EBZ.
Reference: Yu X, Zhang M, Kyker K, Patterson E, Benovic JL, Kem DC. Ischemic inactivation of G protein-
coupled receptor kinase and altered desensitization of canine cardiac -adrenergic receptors. Circulation 102:2535-
2540, 2000.

BASIC POSTER: Signal Transduction II (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                         31
                                                                       Signal Transduction II
P2-58 News Summary
Arthritis drug shows promise in preventing sudden cardiac death after heart attack

An anti-inflammatory drug, normally used to treat arthritis, reduces sudden cardiac death (SCD)
in dogs, through its action of replacing an important protective enzyme that disappears during a
heart attack, according to a new study being presented on Friday, June 20, at The Endocrine
Society’s 85th Annual Meeting in Philadelphia. The researchers hope that these results will lead
to a new approach of preventing sudden death after heart attacks in people.

Dr. Xichun Yu and colleagues from the University of Oklahoma Health Sciences Center in
Oklahoma City have discovered that an enzyme beta adrenergic receptor kinase (BARK),
normally present in the heart muscle, disappears by six hours after a heart attack and is absent for
at least 96 hours. This enzyme normally functions to limit the dangerous effects of increased
adrenaline hormones, such as during fright or stress. If not for its disappearance, it could be
protective against the rapid, irregular rhythms that these hormones can provoke during the first
24–36 hours after a heart attack. The irregular rhythms can lead to sudden cardiac death (SCD),
which represents up to 25 percent of heart attacks in people as well as in dog models of human
heart attack.

In the current study, these researchers examined the effect of inhibiting inflammation, using the
anti-inflammatory drug etanercept, on a dog model of heart attack. There was theoretical reason
to believe it might be protective against loss of BARK, explain the researchers. This drug is
used clinically for treatment of inflammatory diseases like rheumatoid arthritis. Pretreatment
with this drug led to an eight-fold protection against loss of BARK, decreased the frequency of
malignant rhythms that lead to SCD and was protective against SCD in the pretreated animals.
This pretreatment also led to the reduction of heart attack size by 50 percent. Further studies will
be needed to demonstrate the absolute protection of the loss of BARK, thereby providing
alternative approaches to preventing SCD following heart attacks.

This study was supported by the Veterans Affairs Medical Center Merit Review, the University
of Oklahoma Health Sciences Center and Immunex/Amgen Inc.

                                              # # #




                                                32
                     Renin, Mineralocorticoids, Pheochromocytoma & Hypertension
P1-516
Prevalence of Primary Aldosteronism Increase According to the Severity of Hypertension Disease.
Lorena M Mosso1, Cristian A Carvajal1, Alexis A Gonzalez1, Joaquin A Montero2, Carlos E Fardella*1. 1Dept of
Endocrinology, Fac of Med, Pontificia Univ Catolica de Chile, Santiago, Chile; 2Intern Med, Fac of Med, Pontificia
Univ Catolica de Chile, Santiago, Chile.

Recent studies in hypertensive populations using the serum aldosterone (SA) to plasma renin activity (PRA) ratio as
screening test have demonstrated a high prevalence of primary aldosteronism (PA). This frequency is clearly higher
than the classically described when hypokalemia is used as the screening method. However, other factors such as the
characteristic of the hypertensive population studied could also influence the prevalence of PA. We studied 609
essential hypertensive (EH) patients which were classified according the Joint national Committee VI (JNC VI) in
three different stages depending on the severity of the hypertensive disease. We measures SA and PRA and
calculated the SA/PRA ratio in all patients. A SA/PRA ratio greater than 25 was defined as being elevated and a
fludrocortisone test was done to confirm the diagnosis of PA. The SA/PRA ratio higher than 25 was detected in
63/609 and the fludrocortisone test confirmed the diagnoses of PA in 37 cases giving a total prevalence of PA of
6.1%. The prevalence of PA according the hypertension stage was: stage 1 = 6/301 cases (1.99%); stage 2 = 15/187
cases (8.02%) and stage 3 = 16/121 cases (13.2%), frequencies that show a statistically significant difference
(p<0.01). PA patients were lightly younger (48.4 ± 10.5 yrs) than the other hypertensives patients (53.6 ± 10.2 yrs),
difference that reach statistical significant. We did not found differences by gender (male/female) between patients
affected by PA (11/26) or EH (221/388). In the 37 patients with PA, the plasma potassium levels were always within
the normal range. CT scans of the adrenal glands showed a bilateral enlargement in 7 cases and an adrenal nodule in
2 cases. In summary we find a high frequency of PA in EH classified in the stage 2 and 3 of the JVC VI. The low
frequency of CT-scan abnormalities associated to the absence of hypokalemia suggests that most PA patients
diagnosed correspond to mild forms of the disease. These data support the need to redefine the concept of PA as a
continuous pathological disorder in which most patients present an attenuated disease and minorities present the
classical clinical picture.
          Supported by FONDECYT 101-1035

CLINICAL POSTER: Renin, Mineralocorticoids, Pheochromocytoma & Hypertension (11:00 AM - 12:00 PM and
2:30 PM - 3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                         33
                  Renin, Mineralocorticoids, Pheochromocytoma & Hypertension

P1-516 News Summary
Form of hypertension is mild in most but shows full characteristics in minorities

New data support the need to redefine the concept of primary aldosteronism (PA) – a rare but
curable form of hypertension – as a continuous pathological disorder in which most patients have
a milder disease and minorities have the standard clinical disorder, according to a new study
being presented on Thursday, June 19, at The Endocrine Society’s 85th Annual Meeting in
Philadelphia.

Primary aldosteronism (PA), often caused by an adrenal tumor or adrenal hyperplasia, is marked
by the increase in the hormone aldosterone, which is released by the adrenal glands. It is part of
the complex mechanism used by the body to regulate blood pressure. Recent studies using the
serum aldosterone (SA) to plasma renin activity (PRA) ratio as screening test in people with
hypertension have demonstrated a high prevalence of primary aldosteronism close to 10 percent.
This frequency is higher than the classically described case in which hypokalemia, a deficiency
of potassium in the blood, is used as the screening method. However, other factors, such as the
severity of the hypertensive disease, could also influence the prevalence of PA.

Researchers in Santiago, Chile, studied 609 essential hypertensive (EH) patients, which were
classified in three different stages, depending on the severity of the hypertensive disease and
according to Joint National Committee VI (JNC VI) categories. SA and PRA were measured
and the SA/PRA ratio was calculated in all patients. A SA/PRA ratio greater than 25 was
defined as being elevated and a fludrocortisone test was done to confirm the diagnosis of PA.
The SA/PRA ratio higher than 25 was detected in 63 of the 609 patients and the fludrocortisone
test confirmed the diagnoses of PA in 37 cases, suggesting a total prevalence of PA in 6.1
percent of those studied. The prevalence of PA according to the hypertension stage was nearly 2
percent in stage 1 (6 of 301 cases), more than 8 percent in stage 2 (15 of 187 cases) and more
than 13 percent in stage 3 (16 of 121 cases).

PA patients were slightly younger (approximately 38–58 years old) than the other hypertensive
patients (approximately 43–63 years old). No differences were found by gender between
patients affected by PA or EH. In the 37 patients with PA, the plasma potassium levels were
always within the normal range. CT scans of the adrenal glands showed a bilateral enlargement
in seven cases and an adrenal nodule in two cases.

The researchers found a high frequency of PA in EH classified patients in the JNC VI stages 2
and 3. The low frequency of CT-scan abnormalities associated with the absence of hypokalemia
suggests that most PA patients diagnosed have mild forms of the disease. Probably PA patients
with the mild disease represent the most common secondary cause of hypertension.

This research was supported by a grant from the Chilean government.

                                              # # #


                                                34
                                                    Renin, Mineralocorticoids & Hypertension
OR6-6
High Rate of Detection of Primary Aldosteronism, Including Surgically Treatable Forms, Associated with the
“Non-Selective” Screening of Hypertensives Attending a Recently Established Hypertension Unit.
Michael Stowasser*1,2, Richard D Gordon2, Thanuja G Gunasekera1, Diane C Cowley1, Gregory J Ward3, Colin
Archibald4, B Mark Smithers5. 1Hypertens Unit, Univ of Queensland Dept of Med, Princess Alexandra Hosp,
Brisbane, Queensland, Australia; 2Hypertens Unit, Univ of Queensland Dept of Med, Greenslopes Hosp, Brisbane,
Queensland, Australia; 3Queensland Hlth Pathol and Scientific Svcs; 4Dept of Radiology; 5Univ of Queensland Dept
of Surg, Princess Alexandra Hosp, Brisbane, Queensland, Australia.

Wide application of the aldosterone/renin ratio (ARR) among hypertensives has revealed primary aldosteronism
(PAL) to be common, with most patients normokalemic. Some investigators, however, have reported aldosterone-
producing adenoma (APA) to be rare among patients so detected. In a newly established Hypertension Unit at
Princess Alexandra Hospital (PAHHU), we used protocols developed by the Greenslopes Hospital Hypertension
Unit (which reports >30% of patients with PAL to have APAs) to diagnose PAL and determine the subtype. All
patients with elevated ARR (measured after correcting hypokalemia, standardising diet, posture and time of day, and
withdrawing interfering medications including beta-blockers, diuretics, dihydropyridine calcium blockers and
angiotensin receptor blockers) underwent fludrocortisone suppression testing (FST) to confirm (or exclude) PAL. In
confirmed PAL, if genetic testing (long-PCR) excluded glucocorticoid-remediable aldosteronism (GRA), adrenal
venous sampling (AVS) was performed in order to differentiate lateralizing from bilateral PAL. This approach
allowed the PAHHU to diagnose, within its first two years, 54 patients (only 7 [13%] hypokalemic) with PAL. All
tested negative for GRA. Of 49 in whom the subtype has been determined by AVS, aldosterone production
lateralized to one adrenal in 15 (31%; only 6 hypokalemic) and was bilateral in 34 (69%; all normokalemic). CT
demonstrated an ipsilateral mass in only 6 of the 15 who lateralized, and a contralateral mass in one. Fourteen
lateralizing patients underwent unilateral adrenalectomy with cure of hypertension in seven and improvement in the
remainder. Of 12 who underwent post-operative FST, nine demonstrated biochemical cure of PAL (three improved).
In 31 with bilateral PAL followed for at least 3 months, hypertension responded to spironolactone (12.5-50mg/d) or
amiloride (2.5-10mg/d), but these patients required more medications than operated patients (mean 2.4+/-0.2 SEM
versus 0.8+/-0.3; P<0.001) to maintain a similar level of control. When performed with careful regard to
confounding factors, application of the ARR to all hypertensives, followed by FST to confirm or exclude PAL and
AVS to determine the subtype, can result in the detection of significant numbers of patients with specifically
treatable or potentially curable hypertension.

CLINICAL ORAL: Renin, Mineralocorticoids & Hypertension (1:00 PM - 2:30 PM)

Presentation Date: 6/19/2003, Time: 2:15 PM; Location: 113 B




                                                        35
                                             Renin, Mineralocorticoids & Hypertension
OR6-6 News Summary
Rare form of hypertension found to be not so uncommon; good implications for treatment

What was formerly thought to be a rare but treatable form of hypertension is found to be much
more common, according to a study being presented on Thursday, June 19, at The Endocrine
Society’s 85th Annual Meeting in Philadelphia. This increased prevalence is good news for some
people with hypertension because, if diagnosed, this disorder can potentially be cured or their
medication use drastically reduced.

Hypertension is the most common, identifiable chronic disorder in Western societies and can
lead to strokes, heart attacks and kidney failure if not adequately controlled. Control often
means lifelong treatment with antihypertensive drugs, which can be costly and cause side effects,
unless a curable cause for the hypertension can be identified and rectified. Until recently, one
potentially curable form of hypertension, known as “primary aldosteronism” or PAL, was
thought to be rare, accounting for no more than 1 percent of patients. For this reason, most
physicians did not look into this diagnosis unless patients were found to have low plasma
potassium levels – a classic feature of PAL – in addition to having hypertension. In this
condition, the adrenal glands produce too much of a salt-retaining hormone, known as
“aldosterone,” leading to hypertension.

In some patients, the problem is restricted to one of the two adrenals (“unilateral PAL”) and can
be cured by the surgical removal of the overactive gland, which usually contains a benign
aldosterone-producing tumor. In others, the use of aldosterone-blocking medications can
markedly improve hypertension control.

Just more than 10 years ago, researchers in Greenslopes Hospital in Brisbane, Australia, found
that by looking for PAL in all of their patients with hypertension – and not just those with low
plasma potassium levels or very severe hypertension – their rate of detection of PAL increased
by 10 times to 50–90 patients each year, accounting for as many as 10 percent of their
hypertensive patients. These researchers have now confirmed their original findings in a new
study, led by Dr. Michael Stowasser, carried out within the Princess Alexandra Hospital. Within
its first two years of having a hypertension unit, they diagnosed 54 patients as having PAL.

Only seven of them had low plasma potassium levels. Of the 49 who have completed diagnostic
testing, almost one-third were found to have unilateral PAL, and surgery has led to a cure or
improvement of hypertension in all of them. The researchers point out that this high rate of
detection depends on a careful, methodical approach to the diagnostic work up. This work up
takes into account factors that can affect hormone test results and includes blood tests rather than
relying on adrenal CT scanning to distinguish unilateral from bilateral PAL.

This study was supported by the University of Queensland, the National Heart Foundation of
Australia and the Sylvia and Charles Viertel Charitable Foundation.

                                               # # #


                                                36
                                                                                       Neuroendocrinology
OR2-1
Circadian Regulation of Neuroendocrine Genes in the Heart.
Shuo-Yen J Lin*1, Fang Cai1, Sara Arab2,3, Tami Martino2,3, Peter Liu1,2,3, Michael J Sole1,2,3, Denise D Belsham1,3.
1
  Physiol, Univ of Toronto; 2Heart and Stroke Richard Lewar Ctr of Excellence; 3Cell and Molec Biol, Univ Hlth
Network, Toronto, ON, Canada.

The heart has been reported to be an endocrine organ, mainly due to the expression of atrial natriuretic peptide.
Intriguingly, other brain peptides, such as gonadotropin-releasing hormone and neuropeptide Y, have also been
found to be expressed in the heart, however their role and significance in heart physiology is not yet understood. We
used Affymetrix high-density oligonucleotide microarrays to assess the expression of 12,488 murine myocardial
genes at three hour intervals, under normal diurnal conditions of light and dark cycling with no experimental
interference. Variation in myocardial genetic activity in the normal heart was considerable, with 1634 genes (~13%
of the genes analyzed) exhibiting statistically significant changes in expression across the diurnal cycle. Some of
the genes analyzed exhibited rhythmic patterns of expression, while others showed a changing pattern across
light:dark transition times. Interestingly a number of these genes have long been associated with neuroendocrine
function, and are primarily expressed in the brain, particularly the hypothalamus. These include the clock genes
from the superchiasmatic nucleus, including period 1 and 2, bmal1, timeless, cryptochrome 1 and 2, and clock. We
have also confirmed expression of primarily neuroendocrine genes in the heart by reverse transcriptase PCR. These
genes include neuropeptide Y, agouti-related peptide, galanin, growth hormone-releasing hormone, gonadotropin-
releasing hormone, proglucagon, proopiomelanocortin, tryptophan hydroxylase, and the long form of the leptin
receptor. Diurnal regulation of galanin, growth hormone-releasing hormone, gonadotropin-releasing hormone,
proglucagon, proopiomelanocortin, and tryptophan hydroxylase was detected. These results indicate that specific
neuroendocrine genes, thought to function primarily in the brain, may also have vital functions in the heart, and
further analysis of their role in cardiovascular endocrinology is warranted.
Support from Canadian Institutes of Health Research, Heart and Stroke Foundation of Ontario.

BASIC ORAL: Neuroendocrinology (1:00 PM - 2:30 PM)

Presentation Date: 6/19/2003, Time: 1:00 PM; Location: 108 B




                                                         37
                                                                           Neuroendocrinology

OR2-1 News Summary
Heart gene expression is different in the day compared to night; finding important for drug
development

Researchers discover that the activity of genes in the heart have different patterns during the day
as compared to the night, according to a new study being presented on Thursday, June 19, at The
Endocrine Society’s 85th Annual Meeting in Philadelphia. They suggest that adequate sleep and
undisturbed circadian cycling is essential to tissue renewal and normal cardiac structure and
function. Night rhythms should be an important component of the response of the heart to drug
therapy in cardiovascular disease, they add.

In mammals, body processes appear to cycle with a 24-hour day–night (circadian or diurnal)
rhythm. This rhythm is controlled by molecular clocks in the brain and other tissues.
Cardiovascular disease appears to follow a diurnal pattern. For example, adverse events, such as
sudden death and heart attacks, are more likely to occur soon after awakening. Shift workers
have been shown to have a reduced life expectancy.

In the current study, Drs. Denise Belsham and Michael Sole and colleagues in Toronto observed
a striking 24-hour variation in the activity of 13 percent of the genes expressed in the heart. The
activity patterns are clearly divided into day and night phases and involve genes critical for cell
growth, structure and metabolism. A few of these genes, encoding neuroendocrine proteins,
have known functions in the brain but their function in the heart is unclear. Therefore, these
researchers postulate that the heart is regulated differentially in the day as compared to the night.

This research would be expected to have tremendously important implications, say Drs.
Belsham, Sole and colleagues, for the understanding of cardiac physiology and the treatment of
cardiovascular disease (e.g., morning versus evening administration of medications, role of
undisturbed sleep for maintaining heart health, effect of shift work or “jet lag” on cardiac
disease). Understanding these pathways may also result in the development of novel therapies
for heart disease.

This research was supported by the Heart and Stroke Foundation of Canada, the Canadian
Institutes of Health Research and the Natural Sciences and Engineering Research Council of
Canada.

                                               # # #




                                                 38
Diabetes
                                                                                                        Diabetes
P1-597
Does Hemodilution Explain Thiazolidinedione-Related Anemia?
M Cecilia Lansang*1, Michael S Gordon1, Deborah A Price1, Norman K Hollenberg1. 1Med and Radiology,
Brigham and Women’s Hosp, Boston, MA.

Thiazolidinediones are increasingly being used in the treatment of diabetes mellitus. A decrease in hemoglobin (Hb)
and hematocrit (Hct) is a recognized side effect of this class of drugs. With reports of weight gain and edema during
treatment with thiazolidinediones, the drop in Hb and Hct has been thought to reflect hemodilution, but this has not
been verified. We hypothesized that if hemodilution were the principle behind this side effect, then two other
measures would be affected: 1) serum protein would decrease proportionately and 2) changes in body composition
as measured by bioelectrical impedance would proportionately reflect fluid retention. Methods: We studied 2 male
and 7 female type 2 diabetic patients aged 35 to 74 years (mean ± SD: 52.9 ± 12.6) as part of a protocol designed to
investigate the effects of rosiglitazone on the renin-angiotensin system and other hormones regulating sodium
homeostasis. Rosiglitazone was given for 3 months; the dose was started at 4 mg qd then increased to bid. Hb, Hct,
and serum proteins were measured. Weight was monitored. Bioelectrical impedance measurements of fat mass, fat-
free mass and body water components were made. Results: The decrease in Hb and Hct was not accompanied by
proportional changes in 2 different measures: serum proteins and bioelectrical impedance measurements.
  Parameters measured during the 3-month treatment with rosiglitazone
Parameter (mean±SD) Baseline 3 months Percent (%) change
Hb (g/dL)                    13.2 ± 0.9 11.8 ± 0.9 - 9.8
Hct (%)                      39.1 ± 3.4 35.3 ± 3.4 - 9.7
Total serum protein (g/dL) 7.6 ± 0.5 7.2 ± 0.2 - 5.2
Serum albumin (g/dL)         4.4 ± 0.5 4.2 ± 0.3 - 4.5
Weight (kg)                  96.5 ± 20.6 98.2 ± 20.0 + 1.8
Fat-free mass (kg)           53.4 ± 13.6 53.8 ± 12.4 + 0.7
Fat mass (kg)                43.1 ± 17.1 44.4 ± 18.2 + 3.0
Total body water (L)         40.4 ± 9.4 41.3 ± 8.4 + 2.2
Intracellular water (L)      22.3 ± 6.6 22.2 ± 5.7 + 0.4
Extracellular water (L)      18.1 ± 3.2 19.1 ± 3.2 + 5.5
Conclusion: By two different indices, plasma volume expansion does not seem to account for the anemia seen
during rosiglitazone treatment. Bone marrow suppression has been cited by others as a possible mechanism(1).
However, the possibility of blood flow redistribution has to be investigated.
          Reference: 1. Hirase N et al. Thiazolidinedione suppresses the expression of erythroid phenotype in
erythroleukemia cell line K562. Leukemia Research 24(5):393-400,2000.
          This study was funded in part by the NIH and by Glaxo Smith Kline.

CLINICAL POSTER: Diabetes (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                         39
                                                                                          Diabetes
P1-597 News Summary
Diabetic drug shows promise in preventing or treating diabetic kidney disease

Based on important observations of changes in blood cells, protein and body weight and
composition, researchers believe that a class of diabetic drugs, thiazolidinediones (TZDs), has
the potential of preventing or treating diabetic kidney disease, according to a new study being
presented on Thursday, June 19, at The Endocrine Society’s 85th Annual Meeting in
Philadelphia.

TZDs are a class of drugs used in diabetes to lower blood sugar. They are especially attractive
since they do not increase the body’s insulin levels, but rather improve insulin use by the
muscles and fat tissue – decreasing the insulin resistance found in diabetes.

One effect of TZDs is a decrease in hematocrit (red blood cell concentration) and hemoglobin
(the part of the red blood cell that carries oxygen). Because weight gain and edema (swelling)
are also side effects of the TZDs, it has been thought that the drop in hematocrit and hemoglobin
is due to dilution from increased body water retention.

Dr. M. Cecilia Lansang and colleagues at Brigham and Women’s Hospital in Boston, Mass.,
studied seven women and two men and gave them the TZD rosiglitazone for three months. They
measured hemoglobin, hematocrit, serum proteins, body weight and body composition, using a
machine called bioelectrical impedance analyzer. If the drop in hemoglobin and hematocrit were
just due to water being retained by the body, then the serum protein would decline, and body
water would increase, proportionately to the decrease in hemoglobin and hematocrit. This was
not the case. The change in serum protein concentration was about half as much as the change in
hemoglobin and hematocrit. Dilution did not seem to explain these changes, which is important,
say the researchers, because there may be other actions of TZDs that can be explored further.

These findings arose from this research group’s ongoing study on TZDs to understand their
mechanisms of salt and fluid regulation, and their possible beneficial effects on diabetic kidney
disease. The renin-angiotensin system (RAS) is a hormone system linked to the development of
diabetic kidney disease; it is also thought to contribute to insulin resistance in diabetes. If TZDs
are able to reverse “insulin resistance” by acting on the RAS, say the researchers, then TZDs
have the potential of preventing or treating diabetic kidney disease.

                                               # # #




                                                 40
                                                                                                         Diabetes
P1-609
Sildenafil (Viagra®)- Therapy Not Only for Erectile Dysfunction in Type I Diabetes Mellitus?
Jutta C Piswanger-Soelkner*1, Heinz F Hammer1, Wolfgang J Schnedl1, Georg Leb1, Rainer W Lipp1. 1Intern Med,
Karl-Franzens-Univ, Graz, Austria.

Insulin-dependent diabetes mellitus (IDDM) is known to impair virility in up to 60 % as well as gastric emptying
rates. Sildenafil is known to improve erectile dysfunction in up to 56 % of patients but its influence on gastric
emptying rates is as yet unknown. A recently published study with animal has shown a metabolic stimulating effect
of Sildenafil on gastric emptying. A recent study showed a temporal sequel of gastric emptying disorders with the
duration of diabetes mellitus from normal to delayed and finally accelerated gastric emptying.
Aim: The study investigated the effect of Sildenafil on the rate of gastric emptying in patients with long-standing
IDDM.
Material and Methods:
Gastric emptying rates of 18 patients ( 10 female, 8 male) with long-standing IDDM (mean duration of diabetes: 30
± 8 yrs) served as baseline and were repeated within 7 to 14 days 30 min after oral administration of 50 mg of
Sildenafil (Viagra®, Pfizer Ltd, UK). GATED-SPECT myocardial perfusion imaging was performed during the
interval of two emptying examinations to exclude severe coronary artery disease. After ingestion of the standardized
semisolid test meal labelled with Tc-99m, gastric emptying was continuously recorded in supine position during 120
min and after 30 min of postprandial walking, recording was resumed during 20 min.
Results:
None of the patients showed a severe coronary artery disease. Five of 8 men reported erectile dysfunction (63 %).
The mean HbA1c was 8,7 ± 0,9%. Gastric emptying rates (mean ± SD) at 90 min without (50 ± 21) and with
Sildenafil ( 67 ± 24) were compared and showed a general delaying effect (p < 0.001).
Nine patients demonstrated normal gastric emptying, three diabetics showed delayed and six patients accelerated
gastric emptying rates without Sildenafil.
In patients with accelerated gastric emptying rates significance under the influence of Sildenafil was demonstrated at
60 minutes and a general delaying effect at all time points.A significant delaying effect was found in patients with
normal gastric emptying rates at all time points.
Conclusion:.
There may be a benefit for patients with long-standing diabetes mellitus with accelerated gastric emptying rates,
those patients with the longest duration of diabetes mellitus and a high incidence of erectile dysfunction .

CLINICAL POSTER: Diabetes (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                         41
                                                                                        Diabetes
P1-609 News Summary
Sildenafil (Viagra®) improves gastric problem as well as impotence in type-1 diabetics

Counteracting two common problems of type-1 diabetes, sildenafil (Viagra®) improves both
erectile dysfunction and normalizes accelerated gastric emptying, according to a new study being
presented on Thursday, June 19, at The Endocrine Society’s 85th Annual Meeting in
Philadelphia.

Long-standing, insulin-dependent diabetes (type-1 diabetes) is known to impair both virility and
the rates of gastric emptying in up to 60 percent of patients. Sildenafil is a therapeutic agent
known to improve impotence, or erectile dysfunction. In diabetic rodents, sildenafil has been
recently shown to normalize delayed gastric emptying rates in these animals. Therefore,
researchers sought to examine the effect of sildenafil on gastric emptying in human diabetics.
Rapid gastric emptying happens when the lower end of the small intestine fills too quickly with
undigested food from the stomach. Delayed gastric emptying is a disorder in which the stomach
takes too long to empty its contents..

Dr. Jutta C. Piswanger-Soelkner and colleagues at Karl-Franzens-University in Graz, Austria,
studied 18 patients (10 female, eight male) with long-standing, type-1 diabetes – with an average
duration of diabetes between 22–38 years. Gastric emptying was continuously recorded in a
supine position for 90 minutes. After a break of 30 minutes for walking, scintigraphic recording
was resumed in a supine position for an additional 20 minutes. The gastric emptying results
served as the baseline for the second examination seven to 14 days later. The second time, the
recording of gastric emptying started 30 minutes after oral application of 50 mg of sildenafil –
the therapeutic dose for erectile dysfunction.

Of the eight male patients studied, five (63 percent) reported erectile dysfunction. In general,
sildenafil had no effect during the first 60 minutes of gastric emptying; but after 90 minutes, a
delay of emptying rates became significant and remained until the end of recording (140 min).
Therefore, the normalization of delayed gastric emptying in diabetic animals by sildenafil was
not confirmed in the current study. In six patients with accelerated gastric emptying, sildenafil
showed a delaying effect after 60 minutes, remaining slow during the whole recording time.

Therefore, the use of sildenafil seems to have two beneficial outcomes. It has been shown to
improve both erectile dysfunction and to normalize accelerated gastric emptying in long-
standing, insulin-dependent diabetics. Only prokinetic drugs, which normalize delayed gastric
emptying rates, are well established in pharmacy. Therefore, this delaying side effect of
sildenafil may be beneficial to reduce accelerated gastric emptying to normal in diabetic patients.
Accelerated gastric emptying is likely to speed up the increase of unexpected blood sugar levels
after meal ingestion, influencing the necessary insulin application. Sildenafil may, therefore,
improve also glycemic control, say researchers. Additional studies are underway to substantiate
this conclusion.

                                              # # #


                                                42
                                                                                                          Diabetes
P1-611
The Use of Telemedicine in the Management of Poorly Controlled Patients with Diabetes Mellitus.
Robert A Vigersky*1, Tyrone Anderson1, Amy Filmore1, Donna Thomas-Wharton1, Lucia Novak1, Irma J Heltzel1,
Deirdre Smith1, Christine Kessler1, Jeanne Clarkson1, Anne Striegel1, Melissa Loughney1, Robert S Galen2.
1
  Endocrinology Svc, Walter Reed Army Med Ctr, Washington, DC; 2Coll of Pharmacy, Univ of Georgia, Athens,
GA.

Diabetes mellitus is a condition that lends itself to the use of telemedicine and technology because its management
relies heavily on the collection, sharing, and interpretation of blood glucose data by the patient and healthcare
provider. We studied whether or not we could improve the blood sugars of poorly controlled diabetics (A1C > 8.0%)
by giving them one of three technologies to communicate their blood sugar results from home to their provider.
Poorly controlled type 1 and 2 diabetic patients were prospectively randomized to “routine care” (n= 45) or
“technology” (n= 47). Patients in the latter group were further randomized to one of three technologies - modem,
WebTV, internet accessible computer - which connected their glucose meter to their provider via a secure password-
protected website maintained by HealthSentry. Patients in the WebTV and computer groups could also review their
own data on the website. “Routine care” patients were seen by their provider as needed but no less than every 3
months and treated according to clinical practice guidelines. The patients in the “technology” group were similarly
seen but were also instructed to download their blood glucose values weekly by the assigned technology. The
HealthSentry software accepts downloads from any brand of glucose meter which it then analyzes and displays in
tabular and graphic formats. Treatment recommendations were then made based on the data. Patients receiving
“usual care” significantly improved their mean (+/- sem) A1c over 6 months (10.05 ± 0.32 % to 8.32 +/- 0.30%; p <
0.01) as did those in the “technology” group (9.55 +/- 0.24% to 7.83 +/- 0.22%; p < 0.01). However, the
“technology” group did not significantly improve more than those receiving “usual care,” nor were there differences
among the “technology” groups. The number of office visits in 6 months was slightly but not significantly fewer in
the “technology” group (2.7+/-0.3 ) compared with “usual care” (3.2+/-0.3). The “technology” group had a higher
number of telephone consultations (8.4+/-0.7 vs. 4.6+/-0.7 ; p < 0.005). There were no differences in lipid profiles or
microalbumin/creatinine ratios after 6 months.
The data indicate that telemedicine techologies do not improve glycemic control above that of “usual care.” The
ability to show any advantage of these technologies may have been limited by the fact that all patients were treated
by endocrinologists and subspecialty-trained nurse practitioners.

CLINICAL POSTER: Diabetes (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                          43
                                                                                         Diabetes
P1-611 News Summary
Web technology connects diabetes patients with doctor for blood sugar monitoring

Patients with diabetes transmitted data on their blood sugar via the Internet and showed
improvement in their sugar control with fewer visits, according to a study being presented on
Thursday, June 19, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

Diabetes is an epidemic in the United States, affecting 6 percent of the population in general and
up to 27 percent of high-risk groups, such as older African-American women. It is one of the
leading causes of blindness, kidney failure and amputations in adults. High blood sugar is
responsible for these devastating complications; reducing these levels can prevent problems.

Therefore, diabetes lends itself to the use of technology because patients with diabetes need to
measure their blood sugar frequently and provide these results to their medical care provider –
usually at the time of their office visit – to get feedback on improving their sugar control. This
often does not happen for a variety of reasons. Patient forget their data sheets or do not bring in
their meter for downloading. Because there are multiple brands of meters, each of which uses a
proprietary software and cable connection, the physician must have multiple programs on his
computer desktop and numerous cables to connect the meter to the computer.

To overcome these obstacles, Dr. Robert A. Vigersky and colleagues at Walter Reed Army
Medical Center in Washington, DC and the College of Pharmacy at the University of Georgia in
Athens connected patients with their providers via the World Wide Web in an attempt to
improve their overall blood sugar control without additional clinic visits. They studied 92
patients with both type-1 and type-2 diabetes whose blood sugars were in poor control. They
were divided into a “routine care” group – 45 patients – and a “technology” group – 47 patients.
These patients had been referred to a specialty clinic and were seen by either endocrinologists or
nurse practitioners who had been trained in the management of patients with diabetes. Patients
in the latter group were asked to download their blood sugars from their glucose meters weekly
to the HealthSentry Web site (www.HealthSentry.net) via their computers, a modem or WebTV.
This Web site is meter “blind,” being capable of accepting and analyzing the data from 12 brands
of glucose meters. The data is displayed in both tabular and graphic form, and the patients were
contacted after downloading their information as deemed necessary to improve their control.

Both groups had clinic visits about every two to three months over a six-month period. The
“routine care” and “technology” groups had marked but equivalent improvement in the their
blood sugar control over six months, but the “technology” group did so with significantly fewer
clinic visits. Even though the effect of the technology may have been mitigated by the fact that
all patients received care by specialists, achieving the improvement with fewer clinic visits in the
“technology” group suggests that this approach may be both cost saving and improve
patient/physician satisfaction by overcoming barriers to care.

This study was supported by the United States Army Medical Research and Materiel Command.
                                            # # #


                                                 44
                                                                                                   Metabolism
P1-563
Insulin-Like Growth Factor-II (IGF-II) Predicts Future Weight Gain in Normal Weight Subjects with Type 2
Diabetes.
Adrian H Heald*1, Simon Anderson1, Martin Gibson1. 1Dept of Endocrinology and Diabetes, Salford Royal Univ
Hosps NHS Trust, Salford, Greater Manchester, United Kingdom.

Introduction
Insulin-like growth factors-I (IGF-I) and -II (IGF-II) are important in the regulation of metabolism and growth. In
transgenic models of IGF-II overexpression, high levels of IGF-II are associated with decreased fat mass. We have
previously reported in normal weight normoglycaemic individuals that low circulating IGF-II predicts future weight
gain. We went on to explore whether such relationships persisted in circumstances of abnormal glucose tolerance.
Method
We studied 174 type 2 diabetes subjects at baseline and follow-up. Anthropometric and metabolic data were
collected. We assessed the association between baseline IGF-II levels and risk of weight gain (> 2.0 kg) at 5 year
follow-up.
Results
At follow-up 69 (39.7%) participants gained more than 2.0 kg body weight. For normal weight subjects (BMI of 25
or less) at baseline, mean IGF-II levels were significantly lower in those who gained > 2kg weight than weight
stable subjects at 510ng/ml (95 % CI 264-756ng/ml) vs 818 (675-961ng/ml), F=7.4, p=0.01). Multivariate ANOVA
indicated that this difference was independent of baseline fasting insulin C-peptide, glucose, IGF-I, treatment and
duration of diabetes (F=3.1, p=0.06). For this subgroup low circulating IGF-II at baseline strongly correlated with
weight gain (Spearman’s rho= -0.62, p=0.01). With increasing weight the relationship was gradually lost so that for
individuals with BMI greater than 30 no relationship between low IGF-II and future weight gain was found.
Conclusion
Our data show that in normal weight type 2 diabetes subjects, baseline IGF-II concentration is inversely related to
future weight gain, strengthening the putative role for IGF-II in regulating fat mass. We propose that IGF-II
measurement has potential utility in this group for targeting such individuals for early intervention. The lack of a
cross-sectional association between IGF-II and indices of adiposity may be due to compensatory changes in IGF-II
with increasing fat mass.

CLINICAL POSTER: Metabolism (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                        45
                                                                                     Metabolism
P1-563 News Summary
Levels of a hormone can predict diabetics at risk for unhealthy weight gain

Low levels of a hormone in normal-weight diabetics predisposes them to future weight gain,
according to a new study being presented on Thursday, June 19, at The Endocrine Society’s 85th
Annual Meeting in Philadelphia. These results suggest that measuring levels of this hormone
may be valuable for targeting type-2 diabetics with normal body mass at diagnosis for earlier
intervention to reduce the risk of life-threatening complications, such as heart disease and stroke.

The prevalence of adult and childhood obesity is increasing, implicating changing environmental
and cultural factors as the predominant determinants. Variations in susceptibility to weight gain
and obesity may also be the result of disorders in appetite regulation and metabolism, which are
largely regulated by genetic and hormonal factors. Insulin-like growth factor I (IGF-I) and II
(IGF-II) are peptide hormones that circulate in the bloodstream and play an important role in the
regulation of metabolism and growth. Both IGF-I and IGF-II are similar in molecular structure
to insulin. Recent research suggests that IGF-II may be associated with lipid metabolism and
body weight regulation.

People with type-2 diabetes, particularly those who are overweight or obese, have a high
probability of premature death from heart disease and strokes. Many treatments for diabetes tend
to cause weight gain and in some people this can be excessive, resulting in greatly increased
health risks. Physicians would like to be able to target individuals who are more likely to gain
weight so that appropriate resources and treatment can be provided for them. For many years,
researchers have been looking for ways of identifying these individuals before weight gain
occurs but have had little success until now.

Dr. Adrian H. Heald and colleagues at Salford Royal University Hospitals NHS Trust in the
United Kingdom had already found that low blood levels of the cell growth factor IGF-II
predispose men and women without diabetes to weight gain and obesity while higher levels
appear to be protective. In this study, these researchers performed a similar analysis to see if the
same was true in people with type-2 diabetes. They recruited 174 men and women between the
ages of 34 and 70 years old. They then assessed the association between levels of IGF-II at
baseline and the risk of weight gain in the future.

After five years, a total of 69 (39.7 percent) of participants had gained more than 2.0kg of body
weight. As before, they found that people with low blood levels of IGF-II were predisposed to
future weight gain, confirming that low IGF-II can pre-date weight gain in diabetic individuals.
The researchers also discovered that this relationship was only true in those who were not obese
at the start of the study, suggesting some compensatory mechanisms may occur once obesity has
developed.

This study was supported by the United Kingdom NHS North-West Research and Development
Directorate.
                                           # # #


                                                 46
                                                                                                  Metabolism
P1-570
Glucose Intolerance and Altered Vascular Compliance in Healthy, Young, Asian Indian Men in the United
States.
Nicole Simon*1, Pawan Kumar1, Mustafa Tiewala1, Samy McFarlane1, Nathaniel Winer1, Harold E Lebovitz1, Mary
Ann Banerji1. 1Med - Endocrinology, SUNY Downstate Med Ctr, Brooklyn, NY.

Introduction Asian Indians have high rates of cardiovascular (CV) disease and type 2 diabetes. Glucose intolerance,
a known risk factor for CV disease, is reported to be 7% in Asian-Indians.(Diab Med 20: 2003). We hypothesized
that antecedent cardiovascular risk factors could be identified early in young immigrant Asian Indians.
Methods We performed a 75-gram 2 hour oral glucose tolerance test and measured plasma glucose and insulin at 0,
30, 60, 90 and 120 minutes as well as blood pressure (BP) and fasting lipids in 44 young healthy “non-obese” (BMI
25.5±3.4 kg/m2) Asian Indian men, age 25-40 years living in urban United States. Seventy percent had a family
history of diabetes. To further delineate early vascular abnormalities, we measured large (C1)  small (C2) artery
compliance using pulse contour analysis (HDI) in a subset of 12 subjects. Data are mean ± SD.
Results Forty-one percent (18/44) had abnormal glucose tolerance (WHO criteria). Glucose intolerant subjects had
higher BMI  WHR than normals (26.7±3.8 vs 24.3±2.7 kg/m2, p<0.02 and 0.89±0.04 vs 0.84±0.04, p<0.002,
respectively). Metabolic parameters were also significantly higher in the glucose intolerant group: fasting plasma
triglycerides and glucose 182±114 vs 115±58 mg/dl, p<0.02  105±13vs 95±6mg/dl, p<0.004; two hour plasma
insulin and glucose were 116±38 vs 66±42 uU/ml, p<0.001 and 171±30 vs 106±18 mg/dl p<0.0001 respectively.
The BP was higher: systolic BP 113±7 vs 108±7, p=0.05, diastolic BP 76±10 vs 70±8 mmHg, p<0.03. Family
history of diabetes did not differ in the two groups (p=0.22).
Small vessel compliance was 9.8±3.2 ml/mm Hgx100 and large artery compliance was 19±5.9 ml/mm Hgx10. Small
artery compliance was inversely correlated with fasting plasma triglyceride (r= - 0.61, p<0.03), but not BP or
glucose tolerance status. Vascular compliance did not correlate with LDL or HDL cholesterol.
Summary The data indicate that 41% of young, “non-obese”, healthy urban Asian Indian men living in the United
States have glucose intolerance that is associated with higher blood pressure, triglycerides and BMI. Increased
triglycerides are linked to decreased vascular compliance and imply altered endothelial function.
Conclusions These metabolic and vascular changes at an early age may explain the high observed rates of
subsequent CV disease in this immigrant population. This high risk population should be targeted for early detection
and intervention to reduce both diabetes and CV disease.

CLINICAL POSTER: Metabolism (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                        47
                                                                                      Metabolism
P1-570 News Summary
Large percentage of Asian Indian men in U.S. have abnormal blood sugar, leading to
diabetes and heart disease

Forty percent of young Asian Indian men have pre-diabetes, or abnormal blood sugar levels,
which predisposes them to developing diabetes, according to a study being presented on
Thursday, June 19, at The Endocrine Society’s 85th Annual Meeting in Philadelphia. Early
detection of these abnormalities and early appropriate intervention, advise the researchers, would
be beneficial to prevent this group from developing cardiovascular disease as they age.

Asian Indians living in the United States are at high risk for developing diabetes and heart
disease. Dr. Nicole Simon and colleagues at the State University of New York Downstate
Medical Center in Brooklyn wanted to determine the possible reasons involved.

They studied 44 healthy, young, non-obese Asian Indian men – between the ages of 25–40 –
working as health care professionals in urban United States. Seventy percent had a family
history of diabetes. They tested their glucose tolerance using a standard two-hour, 75 gram oral
glucose tolerance test and also measured various types of blood fats – including cholesterol and
triglycerides, blood pressure, weight and height.

In a subset of 12 men, the researchers measured the elasticity of the blood vessels using a non-
invasive method applied over an artery in the wrist. The elasticity of the arteries is a marker of
the blood vessels health. They found that 18 of 44 participants, 41 percent, had abnormal
glucose levels and were pre-diabetic. This group was more obese than the group with normal
blood sugar, had a higher blood pressure and higher levels of triglycerides. People with high
triglycerides had stiffer and less elastic blood vessels than those with lower triglycerides levels.

The researchers found an extraordinary high rate of abnormal glucose metabolism in young
professional Asian Indian men. Impaired glucose tolerance is a known risk factor for
cardiovascular disease. Moreover, Asian Indian men with higher triglycerides level had stiffer
blood vessels than those with normal fat levels.

                                               # # #




                                                 48
                                                                                Diabetes & Metabolism I
P1-284
American Ginseng Prevents IL-1 Induced  Cell Apoptosis and Promotes the Viability of Insulin-Productive
 Cells.
John ZQ Luo1, Naohiro Yano1, LuGuang Luo*1. 1The Hallet Ctr for Diabetes and Endocrinology, Brown Med Sch,
Rhode Island Hosp, Providence, RI.

Ginseng root has been used for millenniums in Asian tonics for its curative and restorative properties. Recently,
ginseng has been found to attenuate hyperglycemia both experimentally and clinically. The loss or disorder of
pancreatic beta cell’s ability to produce insulin and utilize glucose is considered as the major cause for diabetes.
Along with damage done to cell ability, over expression of “programmed cell death” in  cells, known as apoptosis,
has also been observed. We hypothesized that ginseng’s ability to normalized hyperglycemia may act as insulin
stimulator and/or anti-apoptotic factor by accessing pancreatic  cell directly. In the present study, we evaluated the
effect of ginseng extract purchased from H.S.U  Co. (Wisconsin) on insulin production and IL-1 induced
apoptosis utilizing rat immortalized  cell line, INS-1. Serum deprived quiescent cells were cultured with and
without IL-1 (200pg/ml) and ginseng extract (dosages of 5µg, 25µg, 250µg in 0.5mL of cell culture) for 24 hours.
Apoptosis, insulin production and  cell number were evaluated by tri-color fluorescence immunohistochemistry
and the activation of apoptotic factors was measured by western blotting quantified with integrated density. Ginseng
significantly increased insulin producing cells (90.0±0.04 % vs. control 40.2±0.02%, p<0.01). It appears the
response is dosage dependent while over dosage (2500µg) seems to have no benefits for  cell. In TUNEL assay,
IL-1 induced apoptosis (69.2±0.07% vs. control 31.3±3% p<0.01) while under IL-1 circumstances ginseng
reversed the apoptosis (22.0±0.3 % vs. IL-1  69.2±0.07%, p<0.01). To understand the mechanism of ginseng in
anti-apoptosis, we tested the effects of ginseng on two apoptosis related molecules, pro-apoptotic caspase-9 and anti-
apoptotic Bcl-2. IL-1 activated caspase-9 (143.1±11.8 % vs. control, p<0.05) and suppressed Bcl-2 (70.2±7.6 % vs.
control, p<0.05) while ginseng reversed effects of IL-1  on caspase-9 (58.6±12.8 % vs. IL-1  p<0.05) and Bcl-2
(109.7±10.1 % vs. IL-1 , p<0.05). This study proved evident for the first time that ginseng has a direct effect on 
cells to promote insulin production and suppress apoptosis. Ginseng contra IL-1 induced apoptosis is related to the
reverse of effects induced by IL-1 on caspase-9 and Bcl-2. This study gave theoretical evidence to the popular
herbal remedy to solidify its potential to be an effective complementary and alternative medicine for diabetic
patients.

BASIC POSTER: Diabetes & Metabolism I (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                         49
                                                                      Diabetes & Metabolism I
P1-284 News Summary
First-time finding opens pathway to future diabetes treatment with American ginseng

American ginseng root directly affects pancreatic beta cells, which may potentially lead to a
treatment for diabetes, according to a study being presented on Thursday, June 19, at The
Endocrine Society’s 85th Annual Meeting in Philadelphia.

There is strong growing interest worldwide in the use of herbal remedies as alternative
treatments for many different diseases. Ginseng, one of the most widely used herbs for ages, is
considered to play a role in carbohydrate metabolism, diabetes and longevity. Both experimental
and clinical evidence have shown that ginseng lowers blood glucose levels, possibly benefiting
diabetes. However, the mechanisms that may explain this effect are not understood.

American ginseng, Panax quinquefolium, is one of five species of ginseng. Only two of these
five, American and Asian ginseng, are known for their curative properties. Touted for increasing
energy and improving memory, Chinese herbalists also prescribe ginseng root to treat problems
of the digestive and pulmonary systems, nervous disorders, diabetes and low sex drive in men.
Although there have been theories that ginseng acts as an insulin stimulator – accessing
pancreatic β cells directly – it had been unproven.

Therefore, Dr. LuGuang Luo and colleagues at Brown Medical School in Providence, R.I., used
an experimental cell culture system to study the potential effectiveness of ginseng in protecting
insulin-producing pancreatic beta cells from inflammatory cell injury and death. The
inflammatory hormone interleukin-beta (IL-1) was used to induce a process of programmed cell
death (apoptosis) in a cultured pancreatic beta cell line (INS-1). β cells were cultured with or
without IL-1β and an aqueous extract of American ginseng at various doses for 24 hours.
Apoptosis and insulin content of cells were determined. The results indicated that ginseng was
able not only to increase significantly the numbers of insulin-producing cells but also was able to
reverse the numbers of IL-1 β-induced apoptotic cells. To understand how ginseng protects
against IL-1 β-                                        -related molecules, pro-apoptotic caspase-9
and anti-apoptotic Bcl-2, were tested with immunoblotting. Pro-apoptosis caspase-9 was
enhanced while anti-apoptotic Bcl-2 was down-regulated by IL-1 β both significantly versus
control. The most encouraging finding was that ginseng was able to reverse the low level of Bcl-
2 induced by IL-1 β to values similar to control cells and neutralize the effect of enhanced
caspase-9 induced by IL-1 β to below the control level.

Although this work does not provide direct evidence for a practical use of ginseng in treating
diabetes, say researchers, it establishes a direction for future studies on the potential protection of
beta cells by American ginseng.

This work was partially supported by Lifespan Development Grant and Hallet Diabetes
Foundation.
                                            # # #




                                                  50
                                                                                   Diabetes & Metabolism
OR29-1
High Dose Salsalate Improves Glucose Metabolism in Humans with Insulin Resistance.
Waleed A Aldhahi*1, Jongsoon Lee1, Gerald I Shulman2, Steven E Shoelson1, Allison B Goldfine1. 1Res, Joslin
Diabetes Ctr, Boston, MA; 2Intern Med, Yale Univ Sch of Med, New Haven, CT.

Our previous studies have implicated inflammation and the IKK/NF-B pathway in the pathogenesis of insulin
resistance and as molecular targets for its reversal. We have also showed high dose aspirin, acting as an IKK
inhibitor, improves insulin resistance in rodents and patients with type 2 diabetes (T2D). While aspirin and salicylate
are equipotent inhibitors of IKK, salicylate is much safer and has fewer side effects. We therefore investigated the
effects of high dose Disalcid (salsalate), a dimer of salicylic acid, on glucose and lipid metabolism in patients with
impaired glucose tolerance (IGT) and T2D. 7 insulin resistant subjects (5/2 DM/IGT; M/F 2/5; age 49±9 years; BMI
32±6 kg/m²; FBS 112±43 mg/dl; HbA1c 6.3±1.7 %) were evaluated before and after 14 days treatment with high
dose salsalate (4.5g/day, orally, divided doses). Subjects received misoprostol 200mg qid throughout the study.
Average salicylate levels were between 19-28 mg/dl (trough/peak). After two weeks of salsalate fasting glucose
(111.7±43 vs 90.6±19 mg/dl, pre vs post, p=0.03), cholesterol (201±49 vs 177±36 mg/dl, p=0.04), triglycerides
(173.9±97 vs 104.6±53 mg/dl, p=0.007) were all reduced, despite no change in weight. Following treatment there
was a 27% increase in glucose infusion rates (364±154 vs 524±158 ml, p=0.001) during hyperinsulinemic-
euglycemic clamps, a 31% improvement in glucose disposal (M) (4.2 ± 1.7 vs 6.0 ± 2.5 mg/kg/min, p=0.006), and
34% reduction in rates of insulin clearance (p=0.01). Baseline and achieved insulin levels (U/ml) during the clamp
were increased by 34% (7.21±3.5 vs 13.7±8, p=0.03) and 39% (183.8±45.5 vs 300±81, p=0.01), respectively.
Salsalate therapy was also accompanied by a 72% increase in AUC for insulin during first 10 minutes of an IVGTT
(124.5±75 vs 316.2±145 mU, p=0.005) and over 180 min (p=0.01). DNA binding capacity of NFB, assessed in
monocyte extracts, decreased after drug treatment by about 65% (p=0.04) and returned toward baseline levels one
week after drug discontinuation. Therapy was well tolerated except for mild to moderate tinitus; liver enzymes and
creatinine remained within normal ranges. We conclude that high-dose salsalate improves both peripheral glucose
disposal and acute insulin secretory responses in patients with T2D and IGT. These data support the hypothesis that
inflammation contributes to the pathogenesis of insulin resistance and that IKK provides a novel target for the
reversal of insulin resistance in T2D.

CLINICAL ORAL: Diabetes & Metabolism (1:00 PM - 2:30 PM)

Presentation Date: 6/21/2003, Time: 1:00 PM; Location: 201 A




                                                          51
                                                                       Diabetes & Metabolism
OR29-1 News Summary
Derivative of aspirin improves insulin resistance in diabetics

A derivative of aspirin improves insulin resistance in diabetics, without the gastrointestinal side
effects of aspirin, according to a new study being presented on Saturday, June 21, at The
Endocrine Society’s 85th Annual Meeting in Philadelphia. In addition, say researchers, the
mechanism by which it works suggests that inflammation may have a role in developing a
resistance to insulin, which is a precursor to diabetes.

Diabetes and obesity have reached epidemic proportions in the United States and worldwide. Dr.
Steven Shoelson and colleagues in Boston and New Haven, Conn., have been particularly
interested in determining molecular links between these conditions in hopes of developing new
treatments for diabetes. Their previous studies suggested that high-dose aspirin reduced insulin
resistance by inhibiting an inflammatory pathway referred to as IKK/NF-kB. Even though the
clinical effects of high-dose aspirin were impressive, the potentially serious, life-threatening side
effect of gastrointestinal bleeding precluded use of this treatment for anything but validation of
the molecular target.

However, salicylate – which is a salt or ester of salicylic acid – is much safer, particularly in the
form of disalcid, a covalent salicylate dimer. In the current study, these researchers investigated
the effects of high-dose disalcid on glucose and lipid metabolism in seven patients (two men and
five women; ages 40–58) with impaired glucose tolerance and type-2 diabetes. They were
evaluated before and after 14 days of treatment with high-dose salsalate. Participants received
misoprostol, a drug used to prevent stomach ulcers common with anti-inflammatory drugs,
throughout the study.

The results demonstrated that disalcid therapy in type-2 diabetes provides marked improvements
in glucose and insulin levels and lipid profiles. These findings, say the researchers, confirm a
role for inflammation in the pathogenesis of insulin resistance and demonstrate that salicylate, an
inhibitor of IKK/NF-kB, can be used to reverse insulin resistance in patients with type-2
diabetes.

The study was supported by the National Institutes of Health.

                                               # # #




                                                 52
Effects of Hormones on Cancer
                                                                      Effects of Hormones on Cancer
P3-424
Breast Cancer Incidence in Australian Women Using Testosterone in Addition to Estrogen Replacement.
Constantine Dimitrakakis1, Robert A Jones2, Aiyi Liu3, Carolyn A Bondy*1. 1Devel Endocrinology Br, Natl Inst of
Child Hlth and Hum Devel, Bethesda, MD; 2Mem Med Ctr, North Adelaide, Australia; 3DESPR, Natl Inst of Child
Hlth and Hum Devel, Rockville, MD.

Androgens exert anti-mammogenic effects in many clinical situations. We have shown that testosterone inhibits
estrogen’s mitogenic effects on the mammary epithelium in rhesus monkeys (FASEB J, 14:1725). From these
observations, we hypothesized that the addition of testosterone to estrogen and estrogen plus progestin replacement
therapy (HRT) might decrease the breast cancer risk associated with these treatments. To begin to investigate this
possibility, we undertook a retrospective analysis of data collected in 511 post-menopausal Caucasian Australian
women taking testosterone with conventional HRT. All study subjects received implants every 5 months in a dose of
50-150 mg testosterone (most commonly 100 mg), in addition to conventional estrogen treatment. The hormone
therapy was administered by a single practitioner at a private menopause clinic in Southern Australia. Breast cancer
status was ascertained by mammography at the beginning of testosterone treatment and biannually thereafter. The
duration of follow-up was 5.7 ± 2.5 years. Within this observation period, 7 breast cancer cases were diagnosed in
study subjects, resulting in a breast cancer incidence of 240 per 100,000 women-years. The age-specific incidence
rates in our study subjects were the same as those reported for the general population in the South Australian Breast
Cancer Registry. However, the rates are much lower than those reported for age- and race-matched women receiving
conventional HRT. For example, Schairer et al report a rate of 440 cases per 100,000 women-years in women
receiving estrogen alone and estrogen plus progestin (JAMA 2000, 283:485) and the Women’s Health Initiative
study reports a rate of 380 per 100,000 women-years, in women receiving estrogen plus progestin (JAMA 2002,
288:321). These preliminary observations suggest that the addition of testosterone to conventional HRT for post-
menopausal women may indeed reduce the HRT induced breast cancer risk, returning thereby the incidence to
normal rates of the order of those observed in the general untreated population.

CLINICAL POSTER: Effects of Hormones on Cancer (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                         53
                                                            Effects of Hormones on Cancer
P3-424 News Summary
Testosterone added to standard HRT may reduce breast cancer

Testosterone therapy along with standard hormone replacement therapy (HRT) in postmenpausal
women may prevent breast cancer, according to a new study being presented on Saturday, June
21, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

Testosterone was originally used in women using estrogen replacement therapy to encourage a
return of their sex drive and improve sexual function. While this function of the hormone
remains effective for this purpose today, over the years, other, perhaps more important,
subjective benefits have been observed that have taken precedence over the effect on libido.
This has primarily included modifying unstable mood and an improving psychological function.
In addition, there have been observations on the objective benefits, such as significantly
improving bone density to prevent osteoporosis. Moreover, testosterone has not adversely
affected the improvement in cholesterol levels induced by estrogen replacement.

Past studies have suggested that estrogen may have a stimulatory effect on the breasts that is both
uncomfortable and potentially worrying in terms of a possible increased risk of breast cancer.
The use of testosterone appears to block this breast stimulation. However, it is unknown whether
this apparent blocking effect might translate into a decreased incidence of breast cancer.

Researchers at the National Institute of Child Health and Human Development in Bethesda, Md.,
led by Dr. Carolyn Bondy, retrospectively analyzed data collected in 511 postmenopausal
Caucasian Australian women taking testosterone with conventional HRT. All study subjects
received implants every five months in a dose of 50–150 mg testosterone (most commonly 100
mg), in addition to conventional estrogen treatment. The hormone therapy was administered by
Dr. Robert A. Jones, a sole practitioner at a private menopause clinic in Southern Australia.
Breast cancer status was assessed by mammography at the beginning of testosterone treatment
and biannually thereafter. The patients were examined over a period of three to eight years.

Within this observation period, seven breast cancer cases were diagnosed, resulting in a breast
cancer incidence of 240 per 100,000 women-years. The age-specific incidence rates were the
same as those reported for the general population in the South Australian Breast Cancer Registry.
However, the rates are much lower than those reported for age- and race-matched women
receiving conventional HRT.

While this conclusion will have to be more rigorously tested, say the researchers, if it is found to
be accurate, it may remove the main objection to the continued use of estrogen replacement
during menopause that many women find advantageous.

This research was supported by a one time, unrestricted grant from Organon for data tabulation.

                                               # # #



                                                 54
                                           Effect of Hormones on Prostate & Other Cancers
P2-429
Dramatic Growth Inhibition of Human Pancreatic Adenocarcinoma Cells by Four Cardiac Hormones.
Brian A Vesely*1, Quentin McAfee1, William R Gower, Jr1, David L Vesely1,2,3. 1Cardiac Hormone Ctr, Univ of
South Florida Med Sch, Tampa, FL; 2Endocrinology, Diabetes & Metab, James A. Haley Veterans Med Ctr, Tampa,
FL; 3Endocrinology, Diabetes & Metab, Univ of South Florida Med Sch, Tampa, FL.

Cardiovascular endocrinology investigations have revealed that hormones made by the heart (atrial natriuretic
peptides, ANPs) help regulate blood pressure, blood volume and to enhance sodium, potassium and water excretion.
The atrial natriuretic peptide (ANP) gene synthesizes four peptide hormones i.e., long acting natriuretic peptide
(LANP), vessel dilator, kaliuretic peptide, and ANP within the heart. ANP has growth regulatory properties in blood
vessels where it inhibits smooth muscle proliferation (hyperplasia) and growth (hypertrophy). The present
investigation was designed to determine if one or more of these hormones can inhibit the growth and/or decrease the
number of cancer cells. Human pancreatic adenocarcinoma cells were chosen for this investigation since this cancer
has the lowest five-year survival (1%) of all cancers. Vessel dilator, LANP, kaliuretic peptide and ANP each at their
1 micromolar concentration decreased the number of pancreatic adenocarcinoma cells in culture by 65%, 47%, 37%,
and 34%, respectively, within 24 hours. At 96 hours the decrease in cancer cells was 70% (P<0.001); 48% (P<0.01),
42% (P<0.05) and 35% (P<0.05), respectively, secondary to vessel dilator, LANP, kaliuretic peptide, and ANP.
These findings are particularly striking since 5-fluorouracil, which was the standard therapy to treat pancreatic
adenocarcinomas for many years, decreases the growth of pancreatic adenocarcinoma cells in culture by only 16%.
The mechanism of decreasing the number of cancer cells includes a 91%, 84%, 86%, and 83% (P<0.001 for each)
inhibition of DNA synthesis in these cancer cells at 24 hours with 1 micromolar of vessel dilator, LANP, kaliuretic
peptide and ANP, respectively. These cardiac hormones did not effect apoptosis, i.e., programmed cell death, of
these cancer cells. Conclusions: 1) This first demonstration that any cardiovascular hormone can decrease the
number of adenocarcinoma cells revealed that not only one but four cardiac hormones significantly decrease the
number of adenocarcinoma cells. 2) With respect to their ability to decrease the number of cancer cells, vessel
dilator > LANP > kaliuretic peptide > atrial natriuretic peptide.

BASIC POSTER: Effect of Hormones on Prostate & Other Cancers (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                         55
                                    Effect of Hormones on Prostate & Other Cancers
P2-429 News Summary
Hormones made in the heart may be future treatment for most deadly form of cancer

A new study has shown for the first time that four peptide hormones – made primarily within the
heart – can dramatically decrease the number of cancer cells in the most common form of
pancreatic cancer, according to a new study being presented on Friday, June 20, at The
Endocrine Society’s 85th Annual Meeting in Philadelphia.

Pancreatic cancer is the most lethal of common cancers, with the average time of survival being
only four months from time of diagnosis. In addition, only 1 percent of people with the
pancreatic cancer are alive in five years with current chemotherapy. Pancreatic adenocarcinoma
is the form of cancer to which most people are referring when they use the term “pancreatic
cancer.” These tumors account for 75 percent of all cancers of the pancreas.

In the new study, Dr. David Vesely and colleagues at the University of South Florida and the
James A. Haley Veterans Medical Center in Tampa, Fla., found that one of these hormones
decreased the number of human pancreatic adenocarcinoma cells by 70 percent in 24 hours of
treatment. Another of these peptide hormones decreased the number of cancer cells by 50
percent within 24 hours. All four of the peptide hormones decreased the number of cancer cells
by at least twice as much as 5-fluorouracil, which has been the standard treatment of pancreatic
adenocarcinomas for many years. The ability of these hormones to decrease the number of
cancer cells was sustained for at least 96 hours.

Dr. Vesely discovered three of the four hormones that were used in the current study. In
addition, Dr. Vesely and colleagues have previously demonstrated that these peptide hormones
help regulate blood pressure and blood volume by their ability to enhance sodium and water
excretion.

The researchers found that the peptide hormones were able to decrease the number of cancer
cells due to a very potent ability to inhibit DNA synthesis within the cancer cells. They add that
it is important to note that the majority of breast, prostate, and colon cancers are
adenocarcinomas – malignant tumors that originate in the lining of glands – with this cancer also
present in the lung and several other tissues.

                                              # # #




                                                56
Gene Expression
                                                                                        Gene Expression I
P1-28
Unliganded Progesterone Receptor A (PR-A) Isoforms Modify the Phenotype of Human Breast Cancer Cells.
Britta M Jacobsen*1, Stephanie A Schittone1, Jennifer K Richer1, Kathryn B Horwitz1. 1Med/Div of Endocrinology,
Univ of Colorado Hlth Scis Ctr, Denver, CO.

Progesterone target tissues express two progesterone receptors (PR): PR-B and N terminally truncated PR-A. PR-B
have a third activation function and are stronger transactivators than PR-A in vitro. Endogenous genes are also
differentially regulated by liganded PR-A vs. PR-B in human breast cancer cells: progesterone-bound PR-B regulate
different gene subsets and many more total genes than PR-A. To determine the role of unliganded PR on gene
expression, PR-negative T47D breast cancer cells were engineered to inducibly express PR-A (Y iA), PR-B (Y iB)
or no PR (Y iNull). Because PR expression is tightly controlled by inducer dose, and the cells are otherwise
genetically identical, they are ideal models to study ligand-independent effects of PR expression on tumor biology.
Microarray studies were performed in cells before and after induction of no PR, PR-A or PR-B. Cells were treated
with vehicle or inducer for 24h, followed by progesterone or vehicle for 6h. Poly (A) + RNA was labeled, and
Affymetrix gene chips were used to identify PR regulated genes. There are four classes of PR regulated genes: I)
genes regulated only by liganded PR; II) genes regulated by unliganded PR and further upregulated by progesterone;
III) genes regulated equally by liganded and unliganded PR; IV) genes regulated only by unliganded PR. Of 154
total genes regulated by PR, 100 are ligand dependent, and 54 are ligand independent. Among the ligand
independent genes, 18 are regulated by both PR, 7 by PR-B only and 29 by PR-A only. This excess regulation by
unliganded PR-A is surprising, since in the presence of ligand, PR-B are much more transcriptionally active than
PR-A. Our data suggest a novel and unexpected function for unliganded PR-A. Unliganded PR-A regulate genes
that encode cell adhesion molecules, growth regulators, DNA repair enzymes and apoptosis regulators. To determine
whether genes regulated by unliganded PR-A affect breast cancer cell biology, we used Y iA cells to measure
adhesion, migration, apoptosis, growth and differentiation, before and after the receptors were switched on. We
show that in the absence of progesterone, PR-A induction renders cells more: 1) adhesive to extracellular matrix; 2)
resistant to apoptosis; and 3) differentiated, than the same cells in their PR-negative state. Measuring the genes
regulated by PR instead of the receptors themselves may provide an accurate method of assessing human breast
tumor PR content and predicting phenotypic behavior.

BASIC POSTER: Gene Expression I (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

                                                                        Presentation Date: Thursday, June 19, 2003




                                                        57
                                                                           Gene Expression I
P1-28 News Summary
Specific proteins that regulate genes may improve diagnosis and treatment of breast cancer
in the future

Researchers found that progesterone receptors (PRs) – which regulate cellular genes in female
reproductive organs and are used as a measured in breast cancer to guide treatment – are
composed of two types that provide more information than the total PR levels, according to a
new study being presented on Friday, June 20, at The Endocrine Society’s 85th Annual Meeting
in Philadelphia. Researchers believe that these two types of PRs will be important in the future
when assessing breast cancer treatment and prognosis.

Progesterone is a hormone that regulates the function of reproductive organs like the breast,
uterus and ovary. It works by binding to PRs, which are proteins that then regulate cellular
genes. Total amounts of PRs are routinely measured in breast cancers for two reasons: First,
PRs indicate whether a tumor will respond to hormone treatments, such as tamoxifen. Second,
presence of PRs in a tumor indicates the likelihood of a “good” prognosis.

However, researchers have found two types of PRs: PR-B, and a smaller form, PR-A. They
have different functions in breast cancers. However, current tests do not discriminate between
the two, despite the fact that their levels can vary widely in tumors, and some tumors have only
one or the other form. Traditionally, PRs are thought to regulate genes only when bound to
progesterone. But researchers have wondered what happens if when there is little or no
progesterone around, such as in postmenopausal women.

To study this issue, researchers at the University of Colorado Health Sciences Center in Denver,
led by Dr. Britta Jacobsen, engineered breast cancer cells in which PRs can be switched “on” or
“off.” They used these cells to study gene regulation and the behavior and appearance of cells
when PR-A are “off,” and then switched “on.” To measure the regulated genes, they used
“chips” that can measure 12,000 human genes in one experiment.

The researchers found that in the presence of progesterone PR-B turn on many more genes than
PR-A. Surprisingly, even in the absence of progesterone, PRs are able to regulate genes, and
PR-A are more effective than PR-B. Among the genes regulated by PR-A in the absence of
progesterone are ones that control tumor-cell growth and death. In addition, when PR-A are
switched “on,” the cells are less sensitive to chemotherapeutic agents like Taxol, and generally
display more “aggressive” behavior – again in the absence of progesterone. The researchers
believe that it is extremely surprising that PR-A can influence genes the absence of the hormone
progesterone.

This study was supported by the National Institutes of Health, the Department of Defense
Research Service Command, the National Foundation for Cancer Research and The Avon
Foundation.

                                             # # #


                                               58
                                                                                          Gene Expression II
OR38-2
The Use of Salivary Glands as a Potential Gene Transfer Target for Gene Therapeutics of Endocrine
Disorders.
Antony Voutetakis*1, Ioannis Bossis1, Marc Kok1, Jianghua Wang1, Corinne M Goldsmith1, Changyu Zheng1,
Amalia Sertedaki2, John A Chiorini1, Peng Loh3, Lynnette K Nieman4, Bruce J Baum1. 1Gene Therapy and
Therapeutics Br, NIDCR,NIH,DHHS, Bethesda, MD; 2Endocrine Unit, First Ped Dept, Athens Univ Med Sch,
Athens, Greece; 3SCN, Lab of Devel Neurobiol, NICHD,NIH,DHHS, Bethesda, MD; 4Ped and Reprod
Endocrinology Br, NICHD,NIH,DHHS, Bethesda, MD.

Inherited monogenetic endocrine disorders are ideal candidates for gene therapy. We have hypothesized that salivary
glands could be an alternative target for gene delivery for these disorders, potentially offering a safer way of vector
administration without limiting the efficiency of hormone production. The mammalian salivary glands have several
features that make them excellent targets for in vivo gene transfer. They are well encapsulated, limiting undesirable
dissemination of the virus beyond the glands. Their excretory duct orifices can be directly and non-surgically
accessed through intra-oral cannulation, a procedure similar to that used in sialography. In addition, the salivary
glands are not vital organs like many other gene therapy targets, such as the liver, reducing the possibility of serious
side effects.
Adeno-associated virus serotype 2 (AAV) vectors are able to infect dividing and non-dividing cells and establish
long-term transgene expression with minimal pathogenicity. Accordingly, AAV vectors are attractive for use in
many gene therapy applications. In the present study, we constructed two recombinant(r)AAV vectors containing
either the cDNA for human growth hormone (hGH) or human erythropoietin (hEPO), respectively, under the control
of the Rous Sarcoma Virus (RSV) promoter. As an additional consideration for safety, a relatively low dose of
rAAV-GH and rAAV-EPO (5x109 particles) was administered to two groups of Balb/c mice (n=10 each) via
cannulation of the submandibular glands. The two groups served as each other’s control. In the rAAV-EPO treated
group, the serum concentration of hEPO increased from zero on day 1 to a median of 24.6 mU/mL at 4 weeks.
Hematocrit (Hct) levels increased over the same time from a median of ~60% to ~80%. No hEPO, or difference in
the Hct levels, was noticed in the rAAV-GH treated group over this time. In the rAAV-GH treated group, the serum
concentration of hGH increased from zero on day 1 to a median of 200 pg/mL on week 4. Conversely, there was no
hGH detectable in the serum of the rAAV-EPO treated group.
Our results strongly suggest that mammalian salivary glands can be used successfully as gene therapy targets to
correct monogenetic endocrine abnormalities.
          Supported by Intramural Research Program, NIH

BASIC ORAL: Gene Expression II (11:15 AM - 12:45 PM)

Presentation Date: 6/22/2003, Time: 11:30 AM; Location: 203 A




                                                          59
                                                                            Gene Expression II
OR38-2 News Summary
Salivary glands may have role in gene therapy for inherited endocrine problems

Salivary glands can be used successfully as gene therapy targets to correct inherited endocrine
abnormalities, according to a study being presented on Sunday, June 22, at The Endocrine
Society’s 85th Annual Meeting in Philadelphia.

Adeno-associated virus serotype 2 (AAV) vectors are widely used in gene therapy applications
because they can establish long-term transgene expression with minimal pathogenicity.
Approximately 80 percent of the population has experienced an uncomplicated AAV infection.
In the current study, Dr. Antonis Voutetakis and colleagues at the National Institutes of Health in
Bethesda, Md., constructed two recombinant (r) AAV vectors, containing either the cDNA for
human growth hormone (hGH) or human erythropoietin (hEPO), respectively. As an additional
consideration for safety, a relatively low dose of rAAV-GH and rAAV-EPO (5x109 particles)
was used. Vector – sequence of genetic material into which a DNA segment has been inserted
and which can be used to stably introduce exogenous genes into an organism – was administered
to two groups of Balb/c mice (n=10 each) via cannulation of the submandibular glands. The two
groups served as each other’s control.

In the rAAV-EPO treated group, the serum concentration of hEPO increased from zero on day
one to a median of 24.6 mU/mL at four weeks. The hEPO was functional because hematocrit
(Hct) levels increased over the same time from a median of approximately 60 percent to 80
percent. No hEPO or difference in the Hct levels was noticed in the rAAV-GH treated group
over this time. In the rAAV-GH treated group, the serum concentration of hGH increased from
zero on day one to a median of 200pg/mL on week four. No hGH was detectable in the blood of
the rAAV-EPO treated group.

The researchers suggest that salivary glands could efficiently produce hormones, after viral gene
delivery, thus offering an alternative and potentially safer gene therapy target in the case of
inherited monogenetic endocrine disorders. Mammalian salivary glands are well-encapsulated
and not vital organs, which limits undesirable dissemination of the virus and reduces the
possibility of serious side effects, respectively. Their excretory duct orifices can be directly and
non-surgically accessed through intra-oral cannulation, a procedure similar to that used in
contrast radiography of salivary glands (sialography).

This study was supported by the National Institute of Dental and Craniofacial Research.

                                               # # #




                                                 60
Glucocorticoids
                                                                                               Glucocorticoids
P2-579
Maternal Consumption of a High Meat, Low Carbohydrate Diet in Late Pregnancy: Relation to Adult
Cortisol Concentrations in the Offspring.
Kirsten Herrick1, David IW Phillips*1, Soraya Haselden1, Mary Campbell-Brown2, Keith M Godfrey1. 1MRC Envir
Epidemiology Unit, Southampton Gen Hosp, Southampton, United Kingdom; 2Dept of Ob and Gyn, Univ of
Glasgow, Glasgow, United Kingdom.

Recent studies have linked an unbalanced high protein maternal diet in late pregnancy with impaired prenatal
growth and raised adult blood pressure in the offspring. A group of women in Motherwell, Scotland, UK had been
advised to eat 1lb. (0.45 kg) of red meat daily during pregnancy and avoid carbohydrate-rich foods; we previously
found raised blood pressure in the adult offspring of those whose mothers’ had reported high meat/fish and low
green vegetable intakes in late pregnancy. Because high protein diets stimulate the hypothalamic-pituitary-adrenal
axis, these findings could be explained if a high protein maternal diet increased maternal cortisol levels and exposed
the fetus to excess cortisol, programming its developing hypothalamic-pituitary-adrenal axis and leading to lifelong
hypersecretion of cortisol. We tested this hypothesis by measuring fasting plasma cortisol concentrations in 251 of
the men and women aged 28-30 years born in Motherwell. Fasting plasma cortisol concentrations were higher in
men and women with a lower body mass index (P<0.0001) and in those who reported vigorous activity in the
previous four weeks (P=0.03) and greater alcohol consumption (P=0.004). Allowing for the effects of gender,
current body mass index, activity and alcohol consumption, cortisol concentrations increased 5.4% per portion of
maternal meat/fish consumption per day (P=0.03), decreased 3.3% per portion of maternal green vegetables
consumption per week (P=0.14) and were 12.2% higher in those born into manual social class families (P=0.03).
This is the first human evidence that an unbalanced diet during pregnancy may program lifelong hypercortisolaemia
in the offspring.
          Supported by Dunhill Medical Trust and the National Institute of Child Development (1 R01 HD41107-
01).

CLINICAL POSTER: Glucocorticoids (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                         61
                                                                                 Glucocorticoids
P2-579 News Summary
Unbalanced diets in pregnant women may affect lifelong health of their children

Pregnant women who eat unbalanced high protein diets can affect the lifelong health of their
children by altering the intensity of their stress response, according to first-of-a-kind findings
being presented on Friday, June 20, at The Endocrine Society’s 85th Annual Meeting in
Philadelphia.

Mothers of children born at Motherwell Maternity Hospital in Scotland, United Kingdom,
around 30 years ago were advised to eat one pound of red meat per day and avoid carbohydrate-
rich foods during pregnancy.

Previous Motherwell studies show that the offspring of mothers who reported greater meat and
fish and lower green vegetable consumption have evidence of raised blood pressure and glucose
intolerance in adult life. Because high protein diets stimulate the hypothalamic-pituitary-adrenal
axis, researchers believed these findings could be explained if a high-protein maternal diet
increased maternal cortisol levels and exposed the fetus to excess cortisol, programming its
developing hypothalamic-pituitary-adrenal axis and leading to lifelong hypersecretion of cortisol.

Dr. David Phillips and colleagues in Glasgow and Southampton, United Kingdom, tested this
hypothesis by measuring fasting plasma cortisol concentrations in 251 men and women between
the ages of 28–30 years old who were born in Motherwell. They found that they have higher
levels of the stress hormone cortisol. They report that is the first human evidence that an
unbalanced diet during pregnancy may program lifelong hypersecretion of cortisol in offspring.
Further studies are needed to confirm these findings, say the researchers, and to understand their
relevance to pregnant women today. They suggest that an unbalanced maternal diet can increase
the mother’s stress hormone levels, exposing the fetus to excess amounts of these hormones and
leading to persisting changes in stress responses.

This study was supported in part by the Dunhill Medical Trust and the National Institute of Child
Development.

                                               # # #




                                                 62
                                                          Glucocorticoids & Cushing’s Syndrome
OR30-2
Glucocorticoid Therapy Increases Event Rates from Cardiovascular Disease; Results from Two
Pharmacoepidemiological Studies of 265,445 Participants.
Brian R Walker*1, Patrick C Souverein2, Li Wei3, Anick Berard4, Toine CG Egberts2, Hubert GM Leufkens2, Cyrus
Cooper5, Tjeerd van Staa2, Thomas M MacDonald3. 1Endocrinology Unit, Sch of Molec and Clin Med, Univ of
Edinburgh, Edinburgh, United Kingdom; 2Dept of Pharmacoepidemiology and Pharmacotherapy, Utrecht Inst for
Pharmaceutical Scis, Utrecht, Netherlands; 3Meds Monitoring Unit, Dept of Clin Pharmacol and Therapeutics, Univ
of Dundee, Dundee, United Kingdom; 4Fac of Pharmacy, Univ of Montreal, Montreal, Canada; 5MRC Envir
Epidemiology Unit, Univ of Southampton, Southampton, United Kingdom.

Glucocorticoids (GCs) induce systemic cardiovascular risk factors (eg hypertension, hyperglycaemia) but may also
have beneficial anti-inflammatory effects in the vessel wall. The net effect of GC therapy on cardiovascular events,
and its contribution to increased cardiovascular risk in inflammatory conditions such as rheumatoid arthritis, has yet
to be examined in studies with sufficient statistical power to compare event rates.
We performed two studies: (i) a population based cohort study in the Tayside region of Scotland to compare
hospitalisation or death from ischemic heart disease, stroke, or heart failure (n=14,519 events) in subjects with
different levels of GC exposure (total n=164,133, age > 40y, surveyed 1993-96); and (ii) a nested case-control study
in the UK General Practice Research Database (GPRD) to compare GC exposure in subjects with (n=50,656) and
without (n=50,656) these cardiovascular events (age >50y, surveyed 1988-97). GC exposure was classified as
‘none’ (Tayside 54%, GPRD 0%), ‘low’ (topical or inhaled GC only; Tayside 35%, GPRD 73%), ‘medium’
(systemic equivalent to <7.5 mg prednisolone daily; Tayside 10%, GPRD 9%), or ‘high’ (systemic equivalent to >
7.5 mg prednisolone daily; Tayside 1%, GPRD 18%). Data are adjusted relative risks or odds ratios with 95%
confidence intervals.
In both studies, there was a dose-dependent association of GC exposure with events: ‘low’ GC exposure had no
effect; ‘medium’ GC exposure was associated with increased rate of heart failure alone in Tayside (RR 1.56, 95% CI
1.33-1.82) and all cardiovascular events in GPRD (OR 1.44, 1.34-1.55); and ‘high’ GC exposure was associated
with increased rate of all cardiovascular events in both studies (Tayside 2.70, 2.33-3.13; GPRD 1.30, 1.21-1.40). In
the ‘high’ dose group, risks were greatest for heart failure, intermediate for ischemic heart disease, and detected for
stroke only in Tayside. The risks of GC exposure were similar in patients with different underlying diseases and not
attenuated by adjustment for use of anti-rheumatic drugs or bronchodilators.
We conclude that supraphysiological doses (>7.5 mg prednisolone equivalent) of GCs are associated with an
increased risk of cardiovascular events which is both statistically and, given the high prevalence of GC
prescriptions, clinically significant. Moreover, conventionally ‘physiological’ doses of GC are associated with a
small increase in cardiovascular risk which may be relevant, eg in hypopituitarism.

CLINICAL ORAL: Glucocorticoids & Cushing’s Syndrome (1:00 PM - 2:30 PM)

Presentation Date: 6/21/2003, Time: 1:15 PM; Location: 204 A




                                                          63
                                                 Glucocorticoids & Cushing’s Syndrome
OR30-2 News Summary
Anti-inflammatory glucocoritcoids increase a person’s risk for heart disease

People taking high doeses of anti-inflammatory glucocorticoids appear to be at an increased risk
for developing heart problems like heart attacks, strokes and heart failure, according to a new
study being presented on Saturday, June 21, at The Endocrine Society’s 85th Annual Meeting in
Philadelphia.

Glucocorticoid steroids are among the most commonly prescribed drugs and are used in the
treatment of inflammatory diseases, including asthma, rheumatoid arthritis and inflammatory
bowel disease. They are highly effective but also have well-known side effects, such as
aggravating obesity, high blood pressure and diabetes, also risk factors for heart disease. It has
long been suspected that glucocorticoids may increase the risk of heart disease, but the
magnitude of the risk and whether there is a threshold dose below which these steroids are safe
for the heart have not been previously examined.

To investigate these issues, researchers from the United Kingdom, the Netherlands, and Canada,
led by Dr. Brian R. Walker, carried out two studies on the topic. In the first study, conducted in
the Tayside region of Scotland, they collected information over a four-year period on drug
prescriptions, hospitalizations and causes of death in more than 164,000 randomly selected
adults, aged 40 years and older. Nearly half received a prescription for a glucocorticoid, such as
a cream or inhaler, and approximately 2 percent received high doses of oral steroids.
Cardiovascular events – such as heart attacks, strokes and, in particular, heart failure – were
more common in the steroid-treated group. The results suggest that in following 100 patients for
10 years 19 would have a cardiovascular event in the non-steroid treated group, but 32 would
have a heart-related event in the steroid-treated group. The risk was highest in those receiving
high oral doses and negligible in the lowest dose group. The risk could not be accounted for by
the underlying disease for which the steroids were being prescribed.

In the second study, similar information was collected over nine years from the U.K.’s General
Practice Research Database. This analysis included more than 50,000 patients – aged 50 years
and older – who had sustained a cardiovascular event and the same number of healthy controls.
The researchers found that patients with heart disease were 30 percent more likely than controls
to have been taking oral doses of glucocorticoids. Again, the risk was greatest for heart failure
and for the highest steroid doses, and the risk was not explained by the underlying disease.

These results suggest that physicians should aggressively treat cardiovascular risk factors in
patients receiving high doses of glucocorticoids and be vigilant for signs of heart failure. The
researchers emphasize the need to devise drugs with the benefits of steroids but without the
adverse side effects.

The research was supported by the Scottish Executive and the British Heart Foundation.

                                               # # #


                                                64
Growth Hormone
                                  Growth Hormone, Cardiovascular Risk & Malignancy
OR20-3
Low-Dose Growth Hormone (GH) as an Adjuvant to Life Style Modifications in the Therapy of Obesity.
Stewart G Albert*1, Arshag D Mooradian1, Angela T Cestia1, Linda S Hackney1. 1Div of Endocrinology, Dept of
Intern Med, St Louis Univ Sch of Med, Saint Louis, MO.

Background: An optimal goal in therapy of obesity is to lose body fat (BF) and retain lean body mass (LBM). GH
is anabolic and lipolytic; obese subjects have reduced GH and insulin-like growth factor-I (IGF-I) levels. Low-dose
(L-D) GH therapy, aimed at normalizing IGF-I, was added to life style modifications in therapy of obesity. The
study was powered for endpoints of weight (Wt) and BF loss. Methods: 59 subjects (36±7 yrs, BMI 36.9±5.0
kg/m²) were randomized in a double-blind, placebo-controlled trial of L-D GH (Genotropin ™) for a 6-month
intervention phase (6-m) followed by 3-month drug-free retention phase (9-m); 40 completed 6-m and 39 completed
9-m. Twenty-three (16w, 7m) self administered GH 200 mcg daily; after 1-month GH was increased to 400 mcg in
(m) and 600 mcg daily in (w). Controls (Con) of 17 subjects (12w, 5m) administered placebo injections. Body
composition was assessed by dual energy x-ray absorptiometry. All received instruction in diet, behavioral, and
exercise modifications starting at baseline (0-m).
Results: Wt decreased in the GH group from 100.4±13.2 at 0-m, to 98.0±15.6 at 6-m (p=0.04), and was sustained
at 98.1±16.6kg at 9-m (p=0.02). Wt loss in Con was not significant. Wt loss due to GH was entirely due to loss of
BF, which was greater in GH vs. Con at 6-m (-3.5±4.1 vs.-1.6±2.7 kg, p=0.0002) and at 9-m (2.8±4.4 vs.1.3±2.8
kg, p=0.002). LBM did not change in either group. GH increased IGF-I from -0.85 standard deviations (SD) (Z-
score) at 0-m, to +0.09 (SD) at 6-m (p=0.0002). IGF-I did not change in Con. The IGF-I (Z-score) at 6-m correlated
with change in BF(r=0.38, p=0.014). HDL cholesterol increased in the GH group from 43±13 at 0-m, to 51±11 at 6-
m (p<0.001) with a return to 46±9 mg/dL at 9-m. HDL was higher in GH vs.Con at 6-m (p=0.02). In both groups,
there were no changes in triglycerides, LDL, VLDL, glucose, insulin, insulin sensitivity (HOMA or QUICKI),
resting energy expenditure, or blood pressure. GH was well tolerated; 1 subject dropped out due to edema.
Conclusions: Normalization of IGF-I levels with L-D GH therapy was well tolerated, and associated with weight
loss of BF, no change in LBM, and improved lipid profile. The weight effects were retained for 3-m after
discontinuation of therapy. L-D GH therapy may be considered as an adjuvant to a clinically necessary weight loss
program.
          This trial was supported by a clinical grant from Pharmacia & Upjohn

CLINICAL ORAL: Growth Hormone, Cardiovascular Risk & Malignancy (1:00 PM - 2:30 PM)

Presentation Date: 6/20/2003, Time: 1:30 PM; Location: Ballroom A




                                                        65
                             Growth Hormone, Cardiovascular Risk & Malignancy

OR20-3 News Summary
Growth hormone helps obese people lose weight and improves cholesterol levels

Obese people who took moderate doses of growth hormone lost weight, composed solely of
body fat, and maintained muscle mass, according to a new study being presented on Friday, June
20, at The Endocrine Society’s 85th Annual Meeting in Philadelphia. In addition, levels of high-
density lipoprotein (HDL) cholesterol – the “good” cholesterol – increased, which may offer
some protection against developing heart disease.

Obesity is a major health risk in the United States and may affect as much as 30 percent of the
population. It is associated with complications of liver, lung and heart disease and increases the
risk for diabetes. Several medicines are approved for weight loss, and researchers believe that
growth hormone holds future potential to join these drugs as weight-loss therapy in the obese.
When people are obese, their bodies re-adjust their hormone balance. One of the hormones to go
down is growth hormone. When growth hormone is low, the body tends to accumulate body fat,
and lose lean body mass like muscle. When people try to lose weight, the low levels of growth
hormone may interfere with their goals. If they lose muscle, they may not have the energy to
exercise. In the past when doctors used growth hormone as a medicine to restore the body
levels, the doses were too high. People had side effects and did not want to continue the therapy.

Therefore, researchers at St. Louis University School of Medicine in St. Louis, Mo., led by Dr.
Stewart G. Albert, wanted to study the effects of providing growth hormone at a more balanced
dose. They screened 59 people between the ages of 20 and 45 (with an average of 36 years) with
obesity, and 39 people completed the study. The participants were about 40 percent over weight,
as calculated as a body mass index of 37. They were randomized to receive either human growth
hormone or a placebo by nightly injection for six months, followed by three months of receiving
no drugs. There were 23 participants (16 women and 7 men) who received the drug, and 17 (12
women and 5 men) received the placebo.

The dose of growth hormone was calculated to replace the normal amount of growth hormone
the body would make if there were no obesity. Body weight, body fat, and resting metabolic
weight were measured. Those who took growth hormone lost total body weight of about 2.4 kg
(about 5.25 pounds). This weight loss was all due to the loss of body fat; they did not lose any
muscle mass. Of the fat loss, most of it was due to loss of fat in the abdominal area. HDL levels
increased in the group taking growth hormone by about 19 percent. There was no change in the
low-density lipoprotein (LDL) cholesterol, the “bad” cholesterol. Only one study participant had
a side effect – swelling – from taking growth hormone, and the drug was discontinued.

The researchers do not know why people lost weight – whether their appetite decreased or their
energy increased because they maintained their muscle mass and were able to exercise.

The research was supported by a clinical grant from Pharmacia.
                                             # # #


                                               66
                                   Growth Hormone, Cardiovascular Risk & Malignancy
OR20-5
Does GH Treatment Increase the Risk To Develop a Malignant Neoplasm? Experience from KIGS.
Patrick Wilton*1, Anders Mattsson1, Kerstin Albertsson-Wikland2, Christopher Cowell3, Anthony Price4, Edward 0
Reiter5, Michael B Ranke6. 1KIGS/KIMS Outcomes Res, Pharmacia AB, Stockholm, Sweden; 2Dept of Peds, Univ
of Gothenburg, Gothenburg, Sweden; 3Inst of Endocrinology, The Children’s Hosp at Westmead, Westmead,
Australia; 4Dept of Peds, Royal Mancherster Children’s Hosp, Manchester, United Kingdom; 5Dept of Peds,
Baystate Med Ctr Children’s Hosp, Springfield, MA; 6Kinderklinik Sek Paed Endokrinology, Klinikum
Schnarrenberg, Tuebingen, Germany.

Based on two cases each of colon cancer and Hodgkin disease in children treated with pituitary growth hormone
Swerdlow et al (The Lancet 2002; 360:273) suggested that there was an increased mortality of neoplasms associated
with GH treatment. The aim of the present study was to analyse the risk of de novo neoplasms in KIGS (Pharmacia
International Growth Database). By January 2003 there were 40 451 non-tumour patients included in KIGS with at
least a baseline and one return visit on GH, idiopathic GHD (n= 22 511), Turner syndrome (TS) (n=4 923),
Langerhans cell histiocytosis (LCH) (n=162), kidney transplanted (n=123) and other diagnoses (n=12 957) and with
a total GH treatment time of 140 816 years of which 110 177 years were in KIGS. There were in total 26 patients
reported with a de novo neoplasm, eight in IGHD, two in organic GHD, five in TS, four in LCH, three in renal
transplant group and four in four other diagnosis. There were four renal carcinomas of which three were in renal
transplants, three lymphomas, three germinomas of which two were in LCH, two acute myelocytic leukemias and 14
other single neoplasms. Standardized incidence ratio for the whole cohort, but patients with LCH or renal transplants
and in three big subgroups has been calculated age, sex and country adjusted and is shown in Table.
Conclusions: There was no evidence of an increased incidence of neoplasm in GH trated patients, but longer
follow-up is recommended.
                                           Frequency, Incidence and SIR
                                                           No of
                    No of patients Years on GH in KIGS                   Frequency % SIR         95% CI 95% CI
                                                           neoplasms
                                                                                         mean low          high
Idiopathic GHD              22 571                  60 190       8             0.04      1.07    0.46      2.11
Turner syndrome              4 923                  15 518       5              0.1      2.92    0.94      6.81
Other etiologies *          12 957                  34 469       6             0.05      1.41    0.51      3.07
Total                       40 451                 110 177      19             0.05      1.41    0.85      2.21
* Patients with LCH and renal transplanted excluded.


CLINICAL ORAL: Growth Hormone, Cardiovascular Risk & Malignancy (1:00 PM - 2:30 PM)

Presentation Date: 6/20/2003, Time: 2:00 PM; Location: Ballroom A




                                                         67
                               Growth Hormone, Cardiovascular Risk & Malignancy

OR20-5 News Summary
Growth hormone use in children does not increase risk of cancer

A database that collects information about growth hormone use among children shows that the
rate of cancer in this group is no greater than that of similar children not taking this therapy,
putting to rest an ongoing concern, according to a new study being presented on Friday, June 20,
at The Endocrine Society’s 85th Annual Meeting in Philadelphia. The researchers believe that
this finding will be of great interest to physicians treating children with growth hormone.

Children and adolescents with diseases that disturb the growth hormone release from the
pituitary gland have been treated with biosynthetic growth hormone since 1985. The hormone is
identical to that produced in the gland; therefore, it is not surprising that very few side effects are
reported. When a child develops cancer, however, researchers have wondered if growth
hormone could encourage faster growth.

KIGS, the Pharmacia International Growth Database, is a database where physicians from all
over the world can send in data on their growth hormone–treated patients, after permission from
the patient and parents. Information, which could identify a specific patient, is never submitted
to the database.

The aim of the current multi-center, international study, led by Dr. Patrick Wilton, was to
evaluate the risk of developing cancer in growth hormone–treated children and adolescents. As
of January 2003, there were data on 40,451 patients in the KIGS database. Patients who had
been treated for cancer before starting growth hormone were not included. The patients were
treated for an average of 2.7 years. Nineteen were diagnosed with a form of cancer, which was
the expected number that would have been diagnosed among children of the same ages without
growth hormone treatment during a similar period of time.

This study was supported by Pharmacia Corporation.

                                                # # #




                                                  68
                                       Growth Hormone, Cardiovascular Risk & Malignancy
OR20-6
Neoplasms Reported during Growth Hormone Replacement in KIMS.
Patrick Wilton*1, Anders Mattsson1, Roger Abs2, Bengt-Ake Bengtsson4, Ulla Feldt-Rasmussen3, Miklos Goth5,
John Monson6, for KIMS International Board. 1KIGS/KIMS Outcomes Res, Pharmacia AB, Stockholm, Sweden;
2
  Univ Hosp, Antwerp, Belgium; 3RigsHosp., Copenhagen, Denmark; 4Sahlgrenska Hosp, Gothenburg, Sweden;
5
  Natl Med Ctr, Budapest, Hungary; 6St Bart.’s Hosp, London, UK
Does growth hormone treatment increase the risk of neoplasia in adult with GHD? The controversial issue of the
role of growth hormone (GH) in neoplasia still exists. The aim of the present study was to analyse the risk of de
novo neoplasms in adult patients with GHD substituted with growth hormone enrolled in KIMS (Pharmacia
International Metabolic Database). By July 2002 there were 6 428 patients (3 281 males) with at least one baseline
and one return visit on GH enrolled in KIMS. Median age was 44.4 years. Total duration on GH in KIMS was 14
073 years. Etiology to GHD was NFPA (n=1 683), non-secreting adenomas (n=1 219), craniopharyngioma (n=765),
other cranial tumours (n=428), idiopathic GHD (n=981) and miscellaneous (n=1 532). 118 neoplasms were reported
in 115 patients. (Table).
The increased standardized incidence ratio found in skin cancers and cranial tumours were most likely because the
patients were under close surveillance, but also that in the largest group of patients in KIMS, pituitary adenomas,
there is an inherent increased risk of neoplasia. The high SIR based on two cases of carcinoid is influenced by an
inherent risk and not associated with GH replacement.
Conclusions: There is no evidence in KIMS that GH replacement in adult GHD increases the risk to develop a
malignant neoplasm. However, longer follow-up is recommended.
                    Standardized Incidence Ratio (SIR) for different types of neoplasm
                               Number of cases        Standard Incidence Ratio 95% Confidence limits
Type of neoplasm               Observed Expected Mean                     Low           High
Basal cell carcinoma           12         3.8         3.2                 1.7           5.6
Prostate cancer                10         4.8         2.1                 0.98          3.9
Lung cancer                    10         9.2         1.09                0.5           2.0
Breast cancer                  8          8.5         0.9                 0.4           1.9
Cranial tumours                8          1.1         7.5                 3.2           14.8
Melanoma                       6          2.1         2.9                 1.06          6.3
Stomach cancer                 4          2.7         1.5                 0.4           3.8
Bladder cancer                 4          3.0         1.3                 0.4           3.4
Non-Hodgkin lymphoma           3          1.9         1.6                 0.3           4.7
Leukemia                       3          1.3         2.4                 0.5           6.9
Pituitary carcinoma            2          0.03        76                  8.5           274
Pancreas cancer                2          1.5         1.4                 0.2           4.9
Carcinoid                      2          0.18        11.3                1.3           40.7
Colon cancer                   1          4.2         0.2                 0             1.3
Rectal cancer                  1          2.9         0.3                 0             1.9
Corpus uteri myosarcoma 1                 1.5         0.7                 0             3.7
All neoplasms (2ndary incl.) 82           64          1.3                 1.0           1.6
All neoplasms but skin excl. 70           61          1.2                 0.9           1.5
All de novo neoplasms          73         64          1.1                 0.9           1.4
SIR not calc. in 2 cases of vocal cord cancer, single cases of kidney carcinoma, bone tumour and ab. smear.

CLINICAL ORAL: Growth Hormone, Cardiovascular Risk & Malignancy (1:00 PM - 2:30 PM)

Presentation Date: 6/20/2003, Time: 2:15 PM; Location: Ballroom A




                                                               69
                              Growth Hormone, Cardiovascular Risk & Malignancy

OR20-6 News Summary
Lifelong use of growth hormone does not appear to increase risk of cancer

A registry of adults taking growth hormone shows that the incidence of cancer is not
significantly higher in this group than in the general population, which answers a question that
has been proposed by researchers, according to a new study being presented on Friday, June 20,
at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

Growth hormone was originally used to treat children with a disease of the pituitary gland, which
normally produces this hormone. Among other symptoms of a growth hormone deficiency,
these children increase very slowly in height and can end up one or two feet below normal height
as adults. Research has shown that growth hormone has to be injected also in adult life to keep a
normal metabolism. Nowadays, ten thousands of adults around the world are treated with
growth hormone.

There are very few side effects reported with biosynthetic growth hormone, which is not
surprising because the compound is an exact copy of the growth hormone produced in normal
pituitary glands. However, in the case of a growth hormone–deficient patient developing cancer,
researchers have questioned if growth hormone could help the cancer to grow faster.

The aim of the current multi-center, international study, led by Dr. Patrick Wilton, was to
investigate whether adults with growth hormone deficiency and treated with biosynthetic
hormone develop symptoms of cancer more often than people without a growth hormone
disease.

Physicians from 28 countries around the world who are treating patients with growth hormone
are sending data, with patient consent, about possible side effects, including cancer, to a database
called KIMS, Pharmacia International Metabolic Database. Information that could identify a
specific patient is not submitted to the database.

As of July 2002, there were data from 6,428 patients collected. The average time on growth
hormone treatment was 2.2 years. A cancer was diagnosed after the first six months of growth
hormone treatment in 82 patients, which was not more than would be diagnosed in the same
number of normal people of the same ages during the same number of years of observation. Two
types of cancers, skin cancers and brain tumors, were, however, found in higher numbers than
expected. The most likely explanation of this is that the patients were on regular follow-up visits
to their doctors, and small cancers with no symptoms were found. In normal people, most
cancers are found when there are symptoms compelling them to see the doctor or through a
standard screening program, such as for breast cancer. The researchers note that a longer follow-
up period is needed to confirm these early conclusions.

This study was supported by Pharmacia Corporation.
                                          # # #


                                                70
                                                                                           Growth Hormones I
P2-344
The Incidence of Malignant Disease and Cardiovascular Morbidity in Hypopituitary Patients with or without
Growth Hormone Replacement Therapy.
Johan Svensson*1, Bengt-Ake Bengtsson1, Thord Rosen1, Anders Oden2, Gudmundur Johannsson1. 1Res Ctr for
Endocrinology and Metab, Sahlgrenska Univ Hosp, Goteborg, Sweden; 2Kungalv, Valler, Romelanda, Sweden.

Background.The effects of GH replacement therapy in adults on overall mortality, cardiovascular morbidity, and the
rate of malignancies are unknown.
Methods. First, a retrospective comparison was performed between 1411 adult hypopituitary adults and the normal
population in terms of fatal and non-fatal morbidity. Secondly, a prospective comparison was made between 289
hypopituitary patients on long-term GH replacement (1443 patient years) and the background population. Patients
with a history of previous acromegaly or Cushing´s disease were excluded in both studies.
Results. In the 1411 hypopituitary patients without GH replacement, overall mortality, as well as the rates of
cerebrovascular events and malignancies, were increased as compared with the normal population. Cancer in colon
and rectum was the most common malignancy in hypopituitary adults without GH therapy. The rate of myocardial
infarctions was increased in hypopituitary women without GH therapy. In the 289 hypopituitary patients on GH
replacement, overall mortality and the rate of malignancies were similar as in the normal population. The rate of
myocardial infarctions was even lower in hypopituitary women on GH therapy than that in the normal population.
There was a tendency that the rate of cerebrovascular events was increased in the hypopituitary adults on GH
therapy as compared with the normal population.
Conclusion. In hypopituitary patients without GH replacement, the rate of all observed malignancies, as well as the
rate of cancer in colon and rectum, were increased. The rate of myocardial infarctions was increased in hypopituitary
women without GH therapy, and reduced in hypopituitary women on GH replacement, as compared with the normal
population. The rate of cerebrovascular events was increased in hypopituitary adults without GH replacement
therapy, and also tended to be increased in hypopituitary adults on GH replacement therapy.

CLINICAL POSTER: Growth Hormones I (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                         71
                                                                        Growth Hormones I
P2-344 News Summary
Growth hormone treatment shows lower risk of cancer and heart problems for people with
pituitary hormone deficiency, strokes still an issue

In patients with low levels of pituitary hormones, those who were treated with growth hormone
had a lower risk of cancer, heart disease and strokes, according to a study being presented on
Friday, June 20, at The Endocrine Society’s 85th Annual Meeting in Philadelphia. Even so, say
researchers, strokes were still slighly increased in those taking growth hormone when compared
to the general population.

Hypopituitarism often causes deficiency of several hormones, including growth hormone. The
effects of growth hormone replacement therapy in adults on overall mortality, cardiovascular
morbidity and the rate of malignancies are unknown. To study this issue, Dr. Johan Svensson
and colleagues in Göteborg and Romelanda, Sweden, conducted a retrospective comparison of
1,411 hypopituitary adults and the normal population in terms of fatal and non-fatal health
problems. In addition, a similar prospective comparison was made between 289 hypopituitary
patients on long-term growth hormone replacement therapy and the general population. Average
length of treatment was 60 months.

The researchers found that hypopituitary patients without growth hormone replacement had an
increased rate of malignancies, with a predominance of colorectal cancer, suggesting that factors
other than growth hormone is of importance for the risk of cancer in this patient group. In
hypopituitary women, growth hormone replacement appeared to be protective against fatal heart
attacks with rapid time courses.

The rate of cerebrovascular events, such as strokes, was increased in hypopituitary adults without
growth hormone replacement therapy and also tended to be increased in hypopituitary adults on
growth hormone replacement therapy, demonstrating that factors other than growth hormone in
the overall treatment are of importance for the outcome of this patient group.

This work was sponsored by the Swedish Medical Research Council.

                                             # # #




                                               72
                                                                                     Growth Hormones II
P3-495
Body Mass Index Determines Evoked GH Responsiveness in 57 Normal Healthy Male Subjects: A Novel
Diagnostic Challenge.
Vivien S Bonert*1,2, Janet D Elashoff3, Rhodora G Enriquez1, Shlomo Melmed1,2. 1Endocrinology/Med, Cedars-
Sinai Med Ctr, Los Angeles, CA; 2UCLA Sch of Med, Los Angeles, CA; 3Biostatistics Core Services, Research
Inst, Cedars- Sinai Med Ctr, Los Angeles, CA.

Normal spontaneous and evoked GH secretion are determined by multiple factors including gender, age, ethnicity
and body mass index (BMI). GH responses to GH secretagogs, as well as spontaneous 24 -hr GH release are
decreased in obesity. However, IGF-1 concentrations are within the age-adjusted normal range, despite GH
insufficiency associated with obesity . Evoked GH levels in obese normal subjects may be as low as those in
panhypopituitary patients with severe GH deficiency (GHD).The mean GH peak after provocative GH stimulation
with growth hormone releasing hormone/arginine (GHRH+ARG) is 70% lower in obese subjects(BMI>30kg/m²)
than in normal individuals(1).
This study evaluates the clinical utility of provocative GH testing for the diagnosis of GHD in normal healthy male
subjects. Peak GH responses to GHRH+ARG were evaluated in fifty-seven healthy normal male subjects aged 20-
60 years, with BMI from 21-40.7. BMI< 25 is defined as normal, 25-29.9 as overweight, >30 as obese.
After an overnight fast, GHRH (1ug/kg iv) and arginine (0.5g/kg iv over 30 mins) were administered to subjects
and GH levels measured at -30, 0, +30, +60, +90, +120 mins. BMI correlated significantly with peak GH response at
30 (Pearson r = -0.50) and 60 minutes (r = -0.59) after GHRH+ARG. Age correlated moderately with both BMI
(r=0.35) and peak GH responses at 30 (r=-0.33) and 60 min(r=-0.39), but did not contribute significantly to the
prediction of GH response after BMI was entered into the regression.Using the maximum of the 30 min and 60 min
GH responses to GHRH+ARG and classifying peak values < 9 ng/ml as abnormal, 35% of subjects with BMI > 25
failed to respond. In contrast only 1 of 20 subjects with normal BMI (< 25) failed to evoke GH.
Normalized IGF-1 values (derived by subtracting the mean for their decade divided by the standard deviation) did
not correlate with BMI (r=0.19).
Conclusion: These results indicate that BMI is a major determinant of evoked GH response. Peak GH responses are
significantly suppressed in normal subjects with even very mildly elevated BMI (i. e. slightly overweight subjects) .
GH provocative testing for diagnosis of GH deficiency in patients with elevated BMI will not accurately distinguish
normal from deficient GH responses. BMI should be measured and evoked GH responses appropriately interpreted
for all adult subjects undergoing GH testing for GH deficiency.
          Supported by Pharmacia
          Reference: 1)Maccario et al (1999) J . Endocrinol. Invest.22:424-429

CLINICAL POSTER: Growth Hormones II (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                         73
                                                                       Growth Hormones II
P3-495 News Summary
Growth hormone test is not reliable measure in overweight and obese

The growth hormone (GH) stimulation test, which measures for appropriate levels of naturally
occurring GH, may not be a reliable predictor of GH deficiency in healthy people with any
amount of elevated body mass index (BMI), according to a new study being presented on
Saturday, June 21, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

Several factors – including, age, ethnic origin and obesity – influence the level GH in the
bloodstream. Individuals with GH resistance or known pituitary disease may not produce
enough GH. In children. this can cause short stature; in adults, GH insufficiency can lead to
changes in muscle mass, cholesterol levels and bone strength.

GH administration is advised only for people who are truly GH deficient. Patients suspected of
producing too little GH can undergo testing by injecting substances that stimulate the pituitary
gland to secrete GH. Evoked GH levels in response to these stimulants are then measured. It has
been shown, however, that obesity can decrease GH levels in the body to values as low as in
patients with pituitary damage.

Dr. Vivien Bonert and colleagues at Cedars-Sinai Medical Center and the UCLA School of
Medicine, both in Los Angeles, designed a study to establish the degree of obesity that would
affect GH levels. Sixty healthy males, with a body mass index (BMI) ranging from 21–40.7
were given GH stimulation tests, with subsequent measurement of GH levels. BMI relates
weight to height. A BMI of less than 25 is defined as normal, 25–29.9 is overweight and greater
than 30 is considered obese.

The researchers found that even a slight increase in weight can markedly decrease the GH
response to stimulation, which they believe should be an important consideration in interpreting
test results. The stimulation test may not be a valid diagnostic predictor of GH deficiency in
healthy people with any BMI elevation because of the incidence of the false test response caused
by being overweight.

This research is supported by Pharmacia.

                                              # # #




                                               74
                                                  Growth Hormone Measurement & Secretion
OR32-5
A Multicenter, Open-Label Study of Pegvisomant in Patients with Acromegaly Converting from Long-Acting
Octreotide.
Peter J Trainer*1, Pamela U Freda2, Michael O Thorner3, Aart J van der Lely4, David L Kleinberg5, Pia Burman6,
Keith E Friend7. 1Dept of Endocrinology, Christie Hosp, Manchester, United Kingdom; 2Dept of Med, Columbia
Univ, Coll of Physicians & Surgeons, New York, NY; 3Dept of Intern Med, Univ of Virginia Hlth Syst,
Charlottesville, VA; 4Dept of Intern Med, Erasmus Med Ctr, Rotterdam, Netherlands; 5Dept of Med, New York
Univ Med Ctr, New York, NY; 6Global Med Affairs, Endocrine Care, Pharmacia Corp, Stockholm, Sweden; 7Global
Med Affairs, Endocrine Care, Pharmacia Corp, Peapack, NJ.

Goals of acromegaly therapy include adequate biochemical control and reducing signs and symptoms. The major
classes of compounds available to achieve these clinical objectives are somatostatin analogues such as octreotide,
and GH receptor antagonists, represented by the recently approved agent, pegvisomant (Somavert®). We report the
efficacy of pegvisomant to decrease total serum IGF-I levels in a cohort of 48 patients with acromegaly (23 women,
median age 46.5, range 21-78) who were on treatment with the long-acting octreotide, Sandostatin LAR® (LAR).
During pegvisomant treatment at each monthly visit, IGF-I was measured (Nichols acid extraction), a sign and
symptoms score (SSS) completed (5 features of acromegaly each rated on a scale of 0 to 8; total score out of 40,
higher score=more symptoms), and ring size measured. All patients initially participated in a placebo-controlled trial
of pegvisomant (limb 1, L1). The patients were then placed on LAR (L2). Thereafter, patients were converted back
to pegvisomant (L3). The minimum duration of each treatment limb was 3 months.
                                      Results (median and range unless stated)
Parameter               L1 – Pegvisomant              L2 – LAR                           L3 – Pegvisomant
                        17.5 mg/d, (5-40)mean: 18.3 30 mg/mo (10-40) mean:               20 mg/d, (5-40) mean: 17.6
Dose
                        mg/d                          28.2mg/mo                          mg/d
Nadir IGF-I**
                        223 (16-472)                  397.5 (41-825) *†                  240 (13-773)*
(ng/mL)
Normal IGF-I (%) 96% (46/48)                          46% (22/48) *†                     88% (42/48)
SSS                     10 (0-34)                     12.5 (2-36) *†                     9.5 (2-37)
Ring size                                             12.5 (6.5-22) †                    12.0 (7-20)
*different from L1, P<0.005, †different from L3, P<0.05; **For L2, minimum value of 2 levels obtained prior to L3
In summary, preliminary results from this cohort of 48 patients treated serially with pegvisomant, LAR, and then
pegvisomant allows several conclusions to be made about key therapeutic objectives. First, the efficacy of
pegvisomant in achieving biochemical control (IGF-I normalization) is high, even in patients refractory to
octreotide. Additionally, serum IGF-I levels were closely correlated with the clinical markers (IGF-I levels rose with
the decline in clinical symptoms and ring size on LAR and then fell with improvement in these markers on
pegvisomant), confirming that the serum IGF-I level is a meaningful biomarker of disease activity.

CLINICAL ORAL: Growth Hormone Measurement & Secretion (1:00 PM - 2:30 PM)

Presentation Date: 6/21/2003, Time: 2:00 PM; Location: Exhibit Hall C




                                                         75
                                          Growth Hormone Measurement & Secretion
OR32-5 News Summary
New drug shows improvements over existing therapy to treat life-shortening disease

In people with a life-shortening disease called acromegaly, a newly approved medication
effectively controlled levels of a hormone that contributes to the disabling symptoms and the
long-term health problems associated with the disease, according to a study being presented on
Saturday, June 21, at The Endocrine Society’s 85th Annual Meeting in Philadelphia. The drug,
pegvisomant (Somavert®), controlled levels of this hormone, insulin-like growth factor I (IGF-I),
even in patients who did not respond to another drug used to treat acromegaly,

Acromegaly is a serious, life-shortening disease triggered by over-secretion of growth hormone,
most often caused by a pituitary tumor. This excess of growth hormone leads to overproduction
of a second hormone, IGF-I, that contributes to the disabling symptoms and the long-term health
problems associated with the disorder. Patients with acromegaly often suffer from headaches,
excessive sweating, soft-tissue swelling, joint disorders and, perhaps most striking, a progressive
coarsening of facial features and enlargement of the hands, feet and jaw. Patients with
acromegaly face a death rate two-to-four times higher than the average person, due to such
serious long-term complications as heart and respiratory disease, diabetes and some forms of
cancer.

Pegvisomant is the first in a new class of medicines called growth hormone receptor antagonists
and the only medicine designed to specifically block the effects of excess growth hormone in
acromegaly. Pegvisomant was approved on March 2003 by the U.S. Food and Drug
Administration for the treatment of acromegaly.

Dr. Peter Trainer and colleagues from multiple institutions studied 48 men and women with
acromegaly. Patients included in the study initially received pegvisomant as part of a previous
clinical trial. Then they were given long-acting octreotide (Sandostatin LAR®) – also used to
treat acromegaly – for at least three months. This study evaluated the efficacy of converting
these patients from long-acting octreotide to pegvisomant. Levels of IGF-I, the severity of five
clinical signs and symptoms and the ring size of the patients were measured once a month. After
six months of pegvisomant treatment, 88 percent of patients had normal levels of IGF-I,
compared with 46 percent of patients in the octreotide phase and 96 percent of patients in the
previous pegvisomant trial. Patients also had significantly fewer signs and symptoms and
smaller ring size during pegvisomant therapy compared to octreotide therapy.

Results of this study, say researchers, also indicate that measuring the levels of IGF-I in the
blood of acromegaly patients provides an accurate marker of disease activity, as these levels
were closely correlated with clinical markers of the disease, such as signs and symptoms and ring
size.

This study was supported by Pharmacia Corporation.

                                              # # #


                                                76
                                                 Growth Hormone Measurement & Secretion
OR32-3
Development of Ultra Sensitive Human Growth Hormone (hGH) Isoform Specific Immunoassays and Their
Use To Detect Doping with Recombinant hGH.
Zida Wu*1, Martin Bidlingmaier1, Alexandra Keller2, Eberhard Keller2, Christian J Strasburger1. 1Intern Med,
Klinikum der Ludwig-Maximilians-Univ, Munich, Bavaria, Germany; 2Klinik fur Kinder und Jugendliche, Univ
Leipzig, Leipzig, Saxonia, Germany.

Immunoassays specific for hGH isoforms have been shown to be useful tools in the detection of doping with
recombinant hGH (rhGH) (1, 2). To improve the approach, ultra sensitive immunoassays specific for 22 kD-, 20 kD-
and pituitary derived-hGH have been established using enhanced chemiluminescence techniques. Combined with
biotinylated high affinity mAbs, streptavidin conjugated to multiple horseradish peroxidase molecules on a dextran
backbone and sensitive substrates, it was possible to quantify hGH at concentrations as low as 1 pg/ml. Linear
working range of these ultra sensitive immunoassays is between 1-1000 pg/ml, intra- and inter-assay CVs are 3.2-
5.8% and 4.5-7.2% respectively.
To test the power of such ultra sensitive immunoassays in doping detection, we used 35 serum samples taken from 7
subjects between 10 and 18 hours after a single injection of rhGH. These samples have low “total hGH”
concentrations (<0.2 ng/ml) as determined by conventional hGH assays and the treatment with rhGH could not be
detected previously. 20 serum samples from 15 normal, untreated subjects with similar concentrations of hGH were
used as controls. Using the ultra sensitive 22 kD- and pituitary derived-hGH specific immuoassays, 6 out of 7
subjects treated with rhGH could be identified. The analytical methods were adapted for the analysis of urine
samples, and preliminary experiments showed that 20 kDa accounted for less than 0.6% of total hGH in urine
samples from rhGH treated patients (n=7), whereas 1.2-4.1% of total hGH were 20 kD hGH in urine samples from
controls (n=8, p<0.01).
          Reference: (1) Detection of doping with human growth hormone, Wu et al., Lancet, 353: 895, 1999.
          (2) Development of human growth hormone (hGH) isoform specific monoclonal antibodies and their use in
          a possible test to detect hGH doping in sports, Wu et al., Proc 84th Meeting of the Endocrine Society, San
          Francisco, P3-527, 2002.
          This study was supported by grants from BISp (Cologne, Germany, VF 0408/03/01) and the IOC
(Lausanne, Switzerland).


CLINICAL ORAL: Growth Hormone Measurement & Secretion (1:00 PM - 2:30 PM)

Presentation Date: 6/21/2003, Time: 1:30 PM; Location: Exhibit Hall C




                                                        77
                                         Growth Hormone Measurement & Secretion
OR32-3 News Summary
Improved method for detecting growth hormone use by athletes may put stop to abuse

A new method to differentiate naturally occurring growth hormone (hGH) from biosynthetic
growth hormone (rhGH), used in doping in sports, is now more sensitive and can potentially be
used to identify rhGH abuse among athletes, according to a new study being presented on
Saturday, June 21, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

Growth hormone has been estimated as one of the most frequently abused drugs in sport.
Although hGH, as produced by the human pituitary gland, is listed as an prohibited class E
substance by the International Olympic Committee (IOC), no official test for the detection of GH
abuse has been implemented to date. Due to the similarities between rhGH and endogenous GH,
differences have been believed to be undetectable.

With the support of the IOC and the German National Institute for Sport Science, Dr. Zida Wu
and colleagues in Munich and Leipzig, Germany, are developing methods to detect the abuse of
human growth hormone for doping in sports. One of their strategies is developing monoclonal
antibodies and immunoassays able to recognize the difference between rhGH and hGH. The
most widely abused form of growth hormone is the pure preparation produced by recombinant
DNA technology. This protein has a molecular mass of 22 kilo Dalton (kDa). Human pituitary
glands, however, produce not only a form, called an isoform, of GH that has identical or very
similar amino acid sequences to those of rhGH but also hGH dimers – two molecules of hGH
complex – and 20 kDa hGH – hGH lacking 15 amino acid residues.

These researchers have developed monoclonal antibodies (mAbs) that can recognize growth
hormone, distinguishing natural from biosynthetic, even though the differences are very minor.
They selected monoclonal antibodies with a high affinity to recombinant 22 kDa hGH but with
low affinity to pituitary-derived hGH. They also selected monoclonal antibodies with a high
affinity to pituitary-derived hGH but low affinity to recombinant 22 kDa hGH, and monoclonal
antibodies that only bind 20 kD GH or hGH dimers. By using these mAbs, it was possible to
establish immunoassays with high discriminatory potency.

In this study, they have developed a new analysis method, with up to a 100-fold increase in
sensitivity. They examined 35 blood samples taken from seven subjects between 20 and 36
hours after a single injection of rhGH. These samples have low “total hGH” concentrations, and
the treatment with rhGH could not be detected previously. Using the ultra sensitive 22 kD- and
pituitary derived-hGH specific immuoassays, six out of seven subjects treated with rhGH could
be identified. Furthermore, the ultrasensitive assays enable them, for the first time, to analyze
the hGH isoforms in urine samples. Preliminary experiments showed that 20 kDa accounted for
less than 0.6 percent of total hGH in urine samples from seven rhGH treated patients, whereas
1.2–4.1 percent of total hGH were 20 kD hGH in urine samples from eight controls.

This research is supported by the IOC and the German National Institute for Sport Science.
                                             # # #


                                               78
                                                                                                         Lipids
P2-223
Growth Hormone-Releasing Peptides as Inhibitors of Fatty Streaks Formation:A New Therapy for
Atherosclerosis.
Huy Ong*1,2, Andre Tremblay3,4, Maria Febbraio5, Roy Silverstein5, Roberta Avallone1,3, Khadija Iken1, Yanfe
Wang1, Kim Bujold1, Annie Demers1, Martin G Sirois6, Romano Deghenghi7, Sylvie Marleau1. 1Fac of Pharmacy,
Univ de Montreal, Montreal, QC, Canada; 2Dept of Pharmacol, Univ de Montreal, Montreal, QC, Canada; 3Ctr de
Recherche, Hosp Ste Justine, Montreal, QC, Canada; 4Dept Gynecol&Ob, Univ de Montreal, Montreal, QC,
Canada; 5Weill Med Coll, Cornell Univ, New York, NY; 6Montreal Heart Inst, Montreal, QC, Canada;
7
  Europeptides, Argenteuil, France.

We have previously reported that hexarelin (HEX), a member of the growth hormone-releasing peptides (GHRPs),
binds to CD36,a scavenger receptor involved in the scavenging of oxidized low density lipoproteins (oxLDL) in
monocytes/macrophages leading to foam cell formation.To determine whether GHRPs, may interfere with CD36
function/ expression in monocytes/macrophages, thereby hampering the uptake of oxLDL by macrophages and foam
cells development, long term studies of the effects of GHRPs administration on atherosclerotic lesion development
were conducted in ApoE null mice, and in ApoE/CD36 double null mice. Methods :Mice were put on a high fat
high cholesterol (HFHC) diet for a period of 12 weeks.and received daily s.c. injections of 0,9% NaCl or either HEX
(100 g/kg) or EP 80317 (300µg/kg). At 18 weeks, mice were anesthetized and cardiac blood withdrawn for plasma
cholesterol profiles analysis. Peritoneal macrophages were collected for measurement of CD36 expression by flow
cytometry analysis and by western blot.as well as for RNA extraction for RT- PCR analysis of PPAR-LXR-
ABCA1. The aortic trees were dissected and the aortic arch was opened longitudinally The aortic lesion area was
evaluated after staining with oil red-O . Results :A significant reduction of lesion area, by 28% and 47%, was
observed following treatment with HEX and EP 80317, in ApoE null mice respectively.The reduced lesion area in
ApoE null mice was associated with a decrease in total plasma cholesterol (31%) and non HDL cholesterol as
compared to controls. HDL cholesterol tended to increase in ApoE null mice by 65% and 73% following treatment
with HEX and EP 80317, respectively. Both GHRPs reduced ox-LDL induced peritoneal macrophage accumulation
by 39% in ApoE null mice. This reduction was paralleled with the increase of the expression of genes PPAR,
LXR and ABCA1. Conclusions :1)GHRPs protect ApoE null mice from developing fatty streaks lesions. 2)The
anti-atherosclerotic effect of GHRPs is mediated through the negative modulation of the expression of CD36 in
macrophages and the increase of expression of genes involved in the cellular cholesterol removal.3).The protective
effect of GHRPs are associated with favourable modulation in plasma lipid profile. GHRPs might be an interesting
alternative in the treatment of atherosclerosis and hypercholesterolemia.

BASIC POSTER: Lipids (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                        79
                                                                                           Lipids
P2-223 News Summary
New family of drugs shows success in preventing atherosclerosis in early studies

Growth hormone–releasing peptides (GHRPs) prevent fatty streaks and atherosclerotic lesions –
precursors to heart disease – from forming in mice, according to a new study being presented on
Friday, June 20, at The Endocrine Society’s 85th Annual Meeting in Philadelphia. This
discovery may open new perspectives for the treatment of atherosclerosis, also known as
hardening of the arteries.

In recent years, the inflammatory component of atherosclerosis has been revealed. This
inflammatory component is associated with the migration of white blood cell subtypes, the
monocytes, into the inner lining of arteries. In the arterial-prone sites, the monocytes are
transformed into resident cells named macrophages and acquire the ability to engulf lipids – fat
or fat-like substances. Lipid-laden macrophages are known as foam cells and are key elements
of the atherosclerotic lesion. Oxidized lipoproteins are the major lipids taken up by macrophages
through their binding to scavenger receptors, called CD36. The GHRPs that have initially
developed as growth hormone secretagogues – which stimulate secretion of growth hormone –
have shown to exert a protective effect in preventing the foam cell formation.

Researchers in Montreal, New York City and Argenteuil, France, led by Dr. Huy Ong, have
uncovered that GHRPs – a family of small synthetic peptides that are ligands of CD36 – exert a
negative modulatory effect on fatty streak formation and lesion development in ApoE-null mice,
a genetic model of atherosclerosis. They arrived at this conclusion by studying the mice, which
were put on a high fat, high cholesterol (HFHC) diet starting at six weeks of age. The mice
received daily injections for 12 weeks of 0.9 percent NaCl (control) or either one of the two
GHRPs, hexarelin (100 μg/kg) and EP80317 (300 μg/kg). At 18 weeks, blood was collected for
plasma lipid profile analysis and aortic lesions were evaluated.

The effect of GHRPs was associated with a decrease in CD36 expression on macrophages and an
increase in the expression of factors involved in the metabolism and flow of cholesterol from the
macrophages. This protective effect also parallels with a favourable modulation of plasma lipid
and cholesterol profiles.

This is the first report on the clear-cut inhibitory effect of GHRPs on fatty streak formation. The
potential use of these small synthetic peptides as antiatherogenic agents alone or in combination
with statins, say the researchers, might offer a unique alternative for the reduction of
atherosclerotic plaque formation, thus reducing the acute clinical complications of
atherosclerosis, including heart attack and stroke.

This study is supported by the Canadian Institutes of Health Research program and by
Europeptides of Argenteuil, France.

                                              # # #



                                                80
Nuclear Receptors
                                  Novel Functions of Orphan & Other Nuclear Receptors
OR40-1
A Functional Atlas of Orphan Nuclear Receptors.
Ronald N Margolis*1, Ronald M Evans2, Bert W O’Malley3. 1Div of Diabetes, Endocrinology, and Metab, Natl Inst
of Diabetes and Digestive and Kidney Dis, Bethesda, MD; 2Gene Expression Lab, Salk Inst for Biol Studies, San
Diego, CA; 3Molec and Cell Biol, Baylor Coll of Med, Houston, TX.

Orphan Nuclear Receptors (ONRs) represent a branch of the Nuclear Receptor superfamily with several shared
characteristics but distinct structures and emerging functional significance. ONRs function as morphogens,
intracellular regulators of metabolism, and as intermediates in biochemical pathways. To develop a deeper
understanding of their function, the NIH (NIDDK, NIA, NCI) and investigators from 5 institutions (Baylor College
of Medicine, Salk Institute, University of Pennsylvania, UT Southwestern, and Duke University) entered into a
consortium agreement to develop a Functional Atlas of Orphan Nuclear Receptors. The overall focus of the ‘Atlas’
will be to elucidate the role(s) played by ONRs in the development of metabolic processes and disorders, including
Type 2 diabetes, obesity, lipid metabolism, cardiovascular disease, as well as in hormone dependent cancers,
processes of aging and xenobiotic metabolism. Specific goals include: 1) Execute research strategies designed to
rapidly and efficiently elucidate those facets of ONR biology most critical to their understanding; 2) Facilitate the
generation of hypotheses, design of experiments and communication of results by scientists active in this field. The
Atlas will: 1) Use emerging and novel methods to elucidate ONR function, including genomic and metabolic
profiling of ONRs and proteomic profiling of ONR coregulators. 2) Develop a web-accessible bioinformatics
resource in which current and emerging information on ONRs will be organized into accessible and “user-mineable”
forms. 3) Pilot projects to explore chemical profiling of ONRs as part of a discovery process for putative ligands,
and other aspects of ONR structure and function. Information about ONRs will be collated and presented through a
public access website known as the Nuclear Receptor Signaling Atlas (Nursa: “http://www.nursa.org”) which will
become a central respository for information about ONRs.
          Supported by NIH grant: U19DK/AG/CA062434.

BASIC ORAL: Novel Functions of Orphan & Other Nuclear Receptors (11:15 AM - 12:45 PM)

Presentation Date: 6/22/2003, Time: 11:15 AM; Location: Lecture Hall




                                                         81
                            Novel Functions of Orphan & Other Nuclear Receptors

OR40-1 News Summary
New research project focuses on receptors involved in obesity, diabetes, cancer and aging

A new research project is set to uncover information about an important class of receptors in the
body, called orphan nuclear receptors, according to a presentation on Sunday, June 22, at The
Endocrine Society’s 85th Annual Meeting in Philadelphia. These receptors have important
relevance to obesity, diabetes, hormone-dependent cancers and processes of aging, and
researchers hope to elucidate the function of these receptors and to open new avenues for
potential drug development.

The Functional Atlas of Orphan Nuclear Receptors represents a broad, multidisciplinary research
effort to discover new information about an important class of receptors in the body. Orphan
nuclear receptors are related to steroid hormone receptors, such as estrogen and thyroid hormone,
but in many instances do not respond to clearly defined hormones. Orphan nuclear receptors
respond to a variety of natural products in the body, such as dietary lipids, products of fat
metabolism, drugs and toxic substances.

The “Atlas” will collate all information about orphan nuclear receptors into databases that will be
accessible to the public via a Web portal, the Nuclear Receptor Signaling Atlas at
www.NURSA.org. The objective of this work is to catalyze further research. Members of the
consortium working on the project include laboratories and investigators from Baylor College of
Medicine, the Salk Institute, Duke University, University of Texas Southwestern Medical Center
and the University of Pennsylvania.

The project is supported by the National Institute of Diabetes and Digestive and Kidney
Diseases, with co-funding from the National Institute on Aging and the National Cancer
Institute.

                                              # # #




                                                82
                                  Novel Functions of Orphan & Other Nuclear Receptors
OR40-2
Structures of the Nuclear Receptor, HNF4a, Identify Fatty Acids as Its Endogenous Ligands.
Karen M Duda*1, Young-In Chi1, Sirano Dhe-Paganon1, Steven E Shoelson1. 1Cell and Molec Physiol, Joslin
Diabetes Ctr, Boston, MA.

Mutations in HNF4a, a nuclear receptor with prominent functions in liver, gut, kidney and pancreatic  cells, are
associated with an autosomal-dominant form of diabetes (MODY1). We recently solved an x-ray crystal structure of
the HNF4a ligand binding domain (LBD) and have now solved a second structure of the LBD bound to coactivator
SRC peptide. Two conformational states were present in each homodimer of the aporeceptor: an open form with 
helix 12 (12) extended and collinear with 10 and a closed form with 12 folded against the body of the domain.
Both molecules in the coactivator-bound dimer adopt the closed configuration. Otherwise, there were no large
differences between structures of apo- and coactivator-bound receptors. Although all crystallizations were without
added ligands, the ligand binding pockets of both apo- and coactivator-bound receptors contained fatty acids. Gas
chromatographic analyses revealed mixtures of fatty acids and a tendency to select subsets of fatty acids from a
larger pool. In all 3 structural states (apo/open, apo/closed, SRC/closed) the carboxylic acid headgroup of the fatty
acids ion pair with the guanidinium group of R226 at one end of the ligand binding pocket; the aliphatic chains fill a
long, narrow channel that is lined with hydrophobic residues. These findings identify fatty acids as the endogenous
ligands for HNF4a and establish a framework for understanding how HNF4a activity is enhanced by ligand binding
and diminished by MODY1 mutations.

BASIC ORAL: Novel Functions of Orphan & Other Nuclear Receptors (11:15 AM - 12:45 PM)

Presentation Date: 6/22/2003, Time: 11:30 AM; Location: Lecture Hall




                                                         83
                             Novel Functions of Orphan & Other Nuclear Receptors

OR40-2 News Summary
Discovery of the function of genetic flaw in diabetes may lead to new drug treatments

Researchers discover the mechanism by which a genetic cause of diabetes functions, providing a
potential target for the development of new drugs to treat diabetes, according to a new study
being presented on Sunday, June 22, at The Endocrine Society’s 85th Annual Meeting in
Philadelphia.

Nuclear receptors are a special class of proteins that are especially useful as drug targets. Some
nuclear receptors bind hormones, such as estrogen, thyroid hormone, cortisol or vitamin D.
Other nuclear receptors bind molecules involved in metabolic processes, such as fatty acids or
related sterols.

Dr. Steven Shoelson and colleagues at the Joslin Diabetes Center in Boston have been working
on the nuclear receptor HNF4, whose hormone or ligand was unknown. Ligands are a group,
ion or molecule coordinated to a central atom or molecule in a complex. The researchers have
been particularly interested in knowing more about HNF4 because it is associated with diabetes
– one of the few known genetic causes of diabetes.

In the current study, they were able to solve the structure of HNF4to determine how it works
and found that its hormone/ligand binding site was occupied by a fatty acid. This
discovery provided information about its natural ligand, and they concluded that HNF4 is a
fatty acid sensor. Fatty acids are known to cause insulin resistance and diminish insulin
secretion, which fits well into the scheme of potential mediators of diabetes and provides a
potential target for the development of new drugs to treat diabetes.

This study was supported by the National Institutes of Health and the American Diabetes
Association.
                                             # # #




                                                84
Obesity
                                                                                                            Obesity
P1-258
Differential Gene Expression in the Hypothalamus of Obese and Lean Humans: A Preliminary Report.
Angelo DelParigi*1, Thomas G Beach2, Grier P Page3, David B Allison3, Antonio Tataranni1. 1Clin Diabetes and
Nutrit Sect, NIDDK-NIH-DHHS, Phoenix, AZ; 2Civin Lab for Neuropathology and Brain Bank, Sun Hlth Res Inst,
Sun City, AZ; 3Sect on Statistical Genet, Dept. of Biostatistics, Clin Nutrit Res Ctr, Univ of Alabama at
Birmingham, Birmingham, AL.

A large body of evidence indicates that the hypothalamus is central in the regulation of energy homeostasis but little
is known about hypothalamic defects that may underlie human obesity. To identify putative common abnormalities,
we used cDNA microarray technology to determine hypothalamic gene expression in 5 obese (2M/3F, age at
death=51.8±21.3 y, BMI=48.5±12.5 kg/m², mean±SD ) and 5 lean (2M/3F, 50.8±17.7 y, 23.0±2.6 kg/m²) humans.
Based on clinical history and neuropathology report, donors were free of cancer, diabetes or neuro-degenerative
diseases and not known to be on any chronic drug treatment at time of death. Five pair-matched (by sex and age at
death) assays were conducted using a human neurobiology array (Clontech Laboratories, Palo Alto, CA) consisting
of 588 genes. Prior to hybridization, the integrity of the RNA samples was verified using bioanalyzer. Each obese
array was normalized to the paired lean array by a normalization coefficient [ (intensity-background)all genes Lean/ 
(intensity-background)all genes Obese]. Only genes with a background-adjusted signal intensity at least 2 fold greater
than background and not affected by signal bleed were called present. According to background-adjusted intensity
ratios (obese/lean), the transcript of the 5-aminolevulinic acid synthase mitochondrial precursor gene (ALASH)
appeared to be down-regulated in all 5 obese compared to lean donors (mean ratio = 0.36). Two other genes,
histidine decarboxylase (HDC) and caspase-10 precursor (CASP10), appeared to be down- (mean ratio = 0.36) and
up-regulated (mean ratio = 4.07) respectively, in 4 out of 5 obese donors. Paired t-tests yielded p values of 0.045,
0.075, and 0.064 respectively, for these 3 genes. Other genes were more significant and had high true positive
posterior probabilities but did not pass Clontech’s quality control indicators for all arrays. Ongoing research is
attempting to determine the relative validity of different statistical inference strategies applied to these data. In
conclusion, we report possible common abnormalities in the gene expression profile of the hypothalamus of obese
subjects. The altered expression levels of ALASH, HDC, and CASP10 may be cause or consequence of obesity.
Further studies are warranted to confirm these preliminary results and decide if these genes should now be
considered candidate susceptibility genes for obesity.

BASIC POSTER: Obesity (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                          85
                                                                                           Obesity
P1-258 News Summary
First-time finding shows that obese people likely have molecular abnormalities in brain

Obesity may be able to be traced to common molecular abnormalities in the hypothalamus of
obese people’s brains, according to first-of-its-kind data being presented on Thursday, June 19,
at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

Obesity is a hereditary disease that affects millions of people. It is disproportionately prevalent
in some ethnic groups, and it has serious health consequences. Although the cause of obesity
remains unknown in the majority of people, there is increasing scientific consensus that people
gain weight because they eat in excess of their daily energy needs – thought to be caused by
abnormalities in the brain.

Dr. Angelo DelParigi and colleagues at the National Institute of Diabetes & Digestive & Kidney
Diseases (NIDDK) in Phoenix, Sun Health Research Institute in Sun City, Ariz., and the
University of Alabama at Birmingham designed a study to find out, for the first time, if the
molecular make-up of certain parts of the brain consistently differs in obese and lean people.

They studied the hypothalamus first because this region of the brain has been shown in animals
to play a major role in the regulation of body weight. Because of the difficulty of collecting
post-mortem brain samples, the researchers were able to complete the experiment in only five
lean and five obese subjects. With such a small number of subjects, they consider the results to
be exploratory.

To identify presumed common molecular abnormalities, they used micro array technology that
allows differences in the expression of a large number of genes – 588 in this experiment – to be
assessed at the same time. The researchers note that this approach generates large amounts of
data that require sophisticated statistical strategies to be analyzed properly. They identified
several molecules of potential interest for understanding the pathophysiology of human obesity.
In fact, some of these molecules have been also linked to the development of obesity in animals.
However, before they can be certain of the significance of these results, they must be
independently reproduced.

This research was suppported by the NIDDK.

                                               # # #




                                                 86
                                                                                                           Obesity
P3-434
Hormonal and Metabolic Responses to Acute Ghrelin Administration in Patients with Eating Disorders or
Simple Obesity.
Silvia Destefanis*1, Fabio Broglio1, Laura Gianotti1, Ruth Rossetto1, Francesco Tassone1, Valeria Mondelli2, Fabio
Lanfranco1, Secondo Fassino2, Giovanni Abbate Daga2, Carlotta Gauna1, Romano Deghenghi1, Ezio Ghigo1. 1Intern
Med, Div of Endocrinology and Metab, Turin, Italy; 2NeuroScis, Div of Psychiatry, Turin, Italy.

Ghrelin, a gastric-derived natural ligand of the GHS-R, strongly stimulates GH secretion but also possesses other
neuroendocrine, metabolic and non endocrine actions. Ghrelin secretion has been reported to reflect energy balance
status and to be inversely related to body mass index (BMI), being reduced in obesity, and elevated in anorexia
nervosa. Notably ghrelin levels have also been reported elevated in patients with bulimia nervosa in spite of normal
BMI. In order to define the endocrine and metabolic effects of ghrelin in conditions of eating disorders, namely
anorexia and bulimia nervosa, and of simple obesity, we studied GH, PRL, ACTH, cortisol, insulin and glucose
levels after acute i.v. ghrelin (1.0 µg/kg) or saline administration in young women with anorexia nervosa of restricter
type, (AN; n:9; BMI: [mean±SEM]:14.7±0.5 kg/m²), bulimia nervosa, (BN; n:6, BMI: 19.2±2.5 kg/m²), simple
obesity (OB; n;6; BMI:36.3±3 kg/m²) and in normal women (NW; n;7; BMI 20.3±0.5 kg/m²) were also studied. In
all the groups ghrelin significantly increased (p<0.01) GH, PRL, ACTH and cortisol levels. However, the GH
response to ghrelin in NW ( AUC: 4009.2±523.5 µg*min/l) was significantly (p<0.05) higher than in OB
(1491.9±458.1 µg*min/l) and in AN (1833.3±558.5 µg*min/l) that, in turn, were similar. The GH response in BN
(4028±297.3 µg*min/l) was similar to that in NW. In all group the PRL, ACTH and cortisol responses to ghrelin
were similar. Ghrelin increased glucose levels in NW as well as in BN and in OB, but not in AN; on the other hand,
it decreased insulin levels in OB but not in AN, BN and NW. In conclusion, anorexia nervosa and obesity, that are
connoted by ghrelin and GH hyper- and hypo-secretion, respectively, show a selective impairment of the
somatotroph responsiveness to ghrelin administration that is not recorded in bulimia nervosa. Peculiarly, acute
ghrelin administration has no impact on insulin and glucose metabolism in anorexia nervosa.

CLINICAL POSTER: Obesity (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                          87
                                                                                         Obesity
P3-434 News Summary
Hormone that controls appetite has varying effects in anorexia, bulimia and obesity

Researchers uncover, for the first time, information about the hormonal and metabolic effects of
ghrelin – a stomach hormone that triggers the desire to eat – in eating disorders resulting in
anorexia nervosa, obesity and bulimia nervosa, according to a new study being presented on
Saturday, June 21, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

The study shows that anorexia nervosa and obesity, that are defined by the over secretion of
ghrelin and under secretion growth hormone (GH), respectively, show an impairment of the GH
responsiveness to ghrelin administration, which is preserved in bulimia. Moreover, the acute
administration of ghrelin has no impact on insulin and glucose metabolism in anorexia.

Ghrelin, a gastric hormone that has been recently discovered as the natural endogenous
“counterpart” of synthetic molecules known as growth hormone (GH) secretagogues, strongly
stimulates GH secretion through the activation of specific receptors. Ghrelin also stimulates
food intake, controls energy balance, and influences glucose metabolism and cardiovascular
actions. Ghrelin secretion has been reported to reflect energy balance and to be inversely related
to body weight, with levels of ghrelin being reduced in the obese and elevated in lean people.

Researchers in Turin, Italy, led by Dr. Silvia Destefanis, evaluated the hormonal and metabolic
effects of acute intravenous ghrelin administration in women with eating disorders. Nine women
with anorexia nervosa, six women with bulimia nervosa, and in six women with simple obesity
were compared with seven normal women. Obese and anorexic women had similar increases in
GH levels, as induced by ghrelin, but their GH levels were significantly lower than those in
normal and bulimic women, who had similar levels. Moreover, ghrelin increased glucose levels
in normal women as well as in those with bulimia or with obesity but not in women with
anorexia. On the other hand, ghrelin decreased insulin levels in women with obesity but not in
those with anorexia, bulimia and in normal subjects.

This study, say the researchers, provides important new insight in the understanding of the
hormonal and metabolic regulatory mechanisms in three very common clinical conditions –
anorexia nervosa, obesity and bulimia nervosa. These data do not immediately translate into
variations in clinical practice and the care of patients; however, the results provide important
preliminary information about the metabolic and hormonal effects of ghrelin and may provide an
important starting point for further research in this field..

The research was supported by the University of Turin, Italy; Eureka Project; MURST, Rome,
Italy; and FSMEM.

                                              # # #




                                                88
                                                                                                            Obesity
P3-453
Meal Composition at Dinner and Postprandial Nutrient Oxidation in Prepubertal Girls: Are They
Contributing Factors to Obesity?
Claudio Maffeis*1, Lorenza Chini1, Yves Schutz2, Alessandra Grezzani1, Rita Piccoli1, Luciano Tato1. 1Dept of
Peds, Univ of Verona, Verona, Italy; 2Dept of Hum Physiol, Univ of Lausanne, Lausanne, Switzerland.

A positive fat balance is the final mechanism leading to fat gain. Several factors promote a larger fat intake than fat
oxidation in humans. We tested the hypothesis that a meal rich in fat at dinner might promote a different
postprandial energy balance and a different spontaneous food intake at breakfast than an isocaloric, isoproteic meal
with a lower fat content in a group of 10 prepubertal girls (age: 9±1 yrs) with varying excess weight (BMI:
24.6±3.0). The postabsorptive resting energy expenditure and the thermic effect of a meal after a low fat (LF: 19%
fat, 69% carbohydrate, 12% protein) or an isocaloric (2.1 MJ) and isoproteic high fat (HF: 50% fat, 38%
carbohydrate, 12% protein) meal were measured by indirect calorimetry. Postprandial energy expenditure was
measured overnight for 10.5 hrs. Each girl repeated the test with a different randomly assigned menu (HF or LF) one
week after the first test.
The thermic effect of the meal was not significantly different after LF or HF meal (134±78 vs 110±109 kJ/10.5h,
p=ns). Carbohydrate oxidation was significantly higher after LF than after HF meal (39±12 vs 29±9 g/10.5h,
p<0.05). Fat oxidation was not significantly different after the two meals (31±9 LF vs 35±9 g/10.5h HF, p=ns).
However, girls oxidized 1.8±0.9 times more fat than ingested with the LF meal vs 0.3±0.3 times that ingested with
the HF meal (p<0.001). On the contrary, carbohydrates ingested at dinner were not completely oxidized overnight:
girls failed to oxidize 57.5±13.4 % of carbohydrate ingested with the LF meal vs 43.9±17.7% of that ingested with
the HF meal (p=0.03). At breakfast, girls ingested spontaneously a similar amount of energy (1.5±0.7 vs 1.5±0.6 MJ,
p=ns) and nutrient (P: 9 vs 10%, C: 68 vs 64%, F 23 vs 26%, p=ns) after HF or LF at dinner.
In conclusion, HF dinner did not stimulate fat oxidation in absolute value and compensatory food intake at breakfast.
10.5 hrs after dinner, total fat oxidation was higher than fat ingested, with a much larger negative fat balance after a
LF meal. Spontaneous energy and nutrient intake at breakfast was similar after LF and HF isocaloric, isoproteic
dinner. Therefore, despite similar energy content, a lower fat intake at dinner may be useful to promote a
postprandial negative fat balance in overweight prepubertal girls.

CLINICAL POSTER: Obesity (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                          89
                                                                                          Obesity
P3-453 News Summary
Low-fat dinner in young, overweight girls promotes the body’s use of fat

Compared to a high-fat dinner, a meal lower in fat promotes an increase in the body’s use of fat
in overweight prepubertal girls, according to a study being presented on Saturday, June 21, at
The Endocrine Society’s 85th Annual Meeting in Philadelphia. This finding, say the researchers,
may have an immediate consequence in the treatment and prevention of childhood obesity,
consisting of nutritional advice to consume a low-fat diet at dinner.

A positive fat balance is the final mechanism leading to fat gain. Several factors promote larger
fat intake than fat oxidation in humans. Dr. Claudio Maffeis and colleagues in Verona, Italy, and
Lausanne, Switzerland, tested the hypothesis that a meal rich in fat at dinner might promote a
different post-meal energy expenditure and a different spontaneous food intake at breakfast than
a meal with the same calories (isocaloric) and protein content (isoproteic) but with a lower fat
content. They studied 10 overweight, prepubertal girls, between the ages of 8–10 years old, with
body mass index (BMI) ranging from 21–27. The content of the meals in the study were
composed of low fat (20 percent fat, 68 percent carbohydrate, 12 percent protein) or an isocaloric
and isoproteic high fat (50 percent fat, 38 percent carbohydrate, 12 percent protein) meal. The
resting energy expenditure, measured after a 12-hour fast, and the increase of energy expenditure
due to the meal after a low-fat or an isocaloric and isoproteic high-fat meal were measured by
indirect calorimetry. Post-meal energy expenditure was measured overnight for 10.5 hours.
Each girl repeated the test with a different randomly assigned menu (high fat or low fat) one
week after the first test.

The increase of energy expenditure due to the meal was not significantly different after the high-
fat or low-fat meal. Carbohydrate oxidation was significantly higher after the low-fat than after
the high-fat meal. Fat oxidation was not significantly different after the two meals. However,
the girls oxidized 1.8 + 0.9 times more fat than ingested with the low-fat meal versus 0.3 + 0.3
times that ingested with the high-fat meal. On the contrary, carbohydrates ingested at dinner
were not completely oxidized overnight: girls failed to oxidize 57 + 13 percent of carbohydrates
ingested with the low-fat meal versus 44 + 18 percent of that ingested with the high-fat meal. At
breakfast, girls ingested spontaneously a similar amount of energy and nutrients after both meals.

A high-fat dinner did not stimulate fat oxidation in absolute value and compensatory food intake
at breakfast. Ten and a half hours after dinner, total fat oxidation was higher than fat ingested in
both meals, but after the low-fat meal the girls oxidized a greater amount of fat than after the
high-fat meal. Spontaneous energy and nutrient intake at breakfast was similar after the low- and
high-fat isocaloric, isoproteic dinners. Therefore, despite similar energy content, a lower fat
intake at dinner may be useful to promote an increase of the body’s use of fat in overweight
prepubertal girls.

This research was supported by the University of Verona, Italy.

                                               # # #


                                                 90
                                                                                                           Obesity
P3-428
Leptin, but Not Adiponectin, Predicts Stroke in Males.
Stefan Soderberg1, Birgitta Stegmayr1, Lars Weinehall2, Goran Hallmans3, Tommy Olsson*1. 1Dept of Med, Umea
Univ Hosp, Umea, Sweden; 2Dept of Epidemiology and Public Hlth, Umea Univ Hosp, Umea, Sweden; 3Dept of
Nutritional Res, Umea Univ Hosp, Umea, Sweden.

Adipocyte-derived hormones like leptin and adiponectin may be important links between obesity and cardiovascular
disease. In this prospective nested case-referent study, we tested whether leptin and adiponectin are risk markers for
a first-ever stroke.
Two hundred seventy six cases with first-ever stroke (234 cases with ischemic and 42 with haemorrhagic stroke)
were identified who, prior to the stroke, had participated in population based health surveys in northern Sweden.
Referents were matched for sex, age, date and type of health survey and geographic region. Blood pressure (BP),
body mass index (BMI), presence of smoking, diabetes and hypertension were recorded and cholesterol was
analysed. Leptin and adiponectin were analysed in stored samples. Risk markers for first-ever stroke were analysed
by conditional logistic regression analysis.
The stroke event occurred on average 4.9 years after the initial survey. Subjects with a future stroke were more
obese and had higher levels of cholesterol and fasting glucose and had a higher frequency of diabetes mellitus and
hypertension. Leptin levels were higher in male subjects with a future stroke (p=0.004) whereas adiponectin did not
differ between cases and referents. Leptin correlated independently to diastolic blood pressure (p=0.003) and
cholesterol (p=0.01) in men, and to postload glucose (p=0.01) in women whereas adiponectin correlated inversely
with postload glucose in both men (p=0.04) and women (r<0.001). Leptin did not associate with adiponectin once
adjusted for obesity. A high leptin level predicted stroke independently in men (ORQ4=3.16; 95%CI: 1.32-7.61) but
not in women. The increased risk was similar for both ischemic and hemorrhagic stroke. Adiponectin did not
associate with stroke. Males with high leptin developed their stroke faster than males with low levels (p=0.0009)
and the time-related effect of leptin was independent of traditional risk factors. An interaction analysis indicated a
positive interaction between high blood pressure and high leptin in men whereas high leptin levels and adiponectin
were antagonists.
A high leptin level is independently associated with both risk for and time to a first-ever stroke in men but not in
women whereas adiponectin does not associate with a future stroke. Leptin may be a key link in the development of
cardiovascular disease in obesity.

CLINICAL POSTER: Obesity (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                         91
                                                                                         Obesity
P3-428 News Summary
High levels of a fat cell hormone is linked to risk of stroke in obese men

The hormone leptin, which is produced in fat cells, may be a link between obesity and the
development of stroke, according to a study being presented on Saturday, June 21, at The
Endocrine Society’s 85th Annual Meeting in Philadelphia.

Researchers have been looking for a link that can explain the association between obesity and the
development of cardiovascular disease, such as heart attacks and stroke. Dr. Tommy Olsson and
colleagues at Umea University Hospital in Umea, Sweden, designed a population-based study to
look at this issue. The studied was conducted in northern Sweden and included 276 cases of
first-time strokes, with twice the number of controls. They found that high levels of leptin,
independent of other risk factors for stroke, was strongly associated with an increased risk for
developing this disease in men but not in women.

The design of the study was a so-called nested case referent study. The samples in which the
hormones were analyzed was taken before the subjects developed the disease, which makes the
finding strong from an epidemiological standpoint, say the researchers. Modulating leptin levels
or leptin effects on target tissues, such as the sympathetic nervous system might decrease the risk
for developing stroke.

The study was supported by the Swedish Medical Research Council and Swedish Heart and
Lung Foundation.

                                              # # #




                                                92
                                                                                                        Obesity
P3-467
High Prevalence of Vitamin D Insufficiency in Obese African American Subjects.
Shamik J Parikh*1, Marni Edelman1, Denkinger Blakeley2, Karim A Calis3, Pamela C Slaughter1, Teresa McHugh1,
Jack A Yanovski1. 1Unit on Growth and Obesity, Develpmental Endocrinology Br, Natl Inst of Child Hlth and
Develpment, Natl Insts of Hlth, Bethesda, MD; 2Nutrit Dept, Clin Ctr, NIH, Bethesda, MD; 3Clin Ctr Pharmacy
Dept, NIH, Bethesda, MD.

Background: Ultraviolet light has previously been shown to be less able to stimulate synthesis of Vitamin D in
African American (AA) than Caucasian (C) adults. Previous studies have also found obesity to be associated with
low 25- hydroxy Vitamin D (25-OH Vit D) and high intact parathyroid hormone (iPTH) levels. Because obesity is
highly prevalent among AA, we hypothesized that Vitamin D deficiency and secondary hyperparathyroidism would
be highly prevalent in obese African American adults.
Methods: Following an overnight fast, we measured serum calcium, phosphorous, iPTH and 25-OH Vit D in 164
healthy overweight (BMI  25) C (n = 95, 55% females) and AA (n = 59, 88% females) adults. We defined Vitamin
D insufficiency as a 25-OH Vit D < 20 ng/dl and hyperparathyroidemia as an iPTH > 65 pg/ml.
Results: There was a significant negative correlation between 25-OH Vit D and BMI (r = - 0.44, p < 0.0001) and a
significant positive correlation between iPTH and BMI (r = 0.38, p < 0.0001). In a multiple regression model, after
including race and gender; 25-OH Vit D was still negatively correlated to BMI (r = -0.35, p = 0.0001) while iPTH
was positively correlated to BMI (r = 0.31, p <0.0001). Compared with C, AA were heavier (AA 36.0±7.5 kg/m² vs.
C 31.2±5.7 kg/m², p<0.0001), had lower 25-OH Vit D (18±8.1 vs. 32±10.7 ng/ml, p<0.0001) and higher iPTH
(60.6±19.7 vs. 51.2±16.6 pg/ml, p=0.0019), but after adjustment for differences in BMI and gender, only 25-OH Vit
D concentrations were different in AA and C (19.2±9.8 vs. 31.0±9.6 ng/ml, p < 0.001). Of the obese (BMI  30) AA
subjects (n = 44, BMI 39.1 ± 6.2 kg/m²), 66% had 25-OH Vit D < 20 ng/ml versus only 5 % of the obese C subjects
(n = 43, BMI 35.9 ± 5.2 kg/m²), p < 0.0001. 48% of the obese AA subjects versus 33% of the obese C subjects had
iPTH  65 pg/ml, (p = 0.19)
Conclusions: Both body mass index and race appear to be important risk factors for Vitamin D insufficiency. As a
result, obese African Americans are at significant risk for Vitamin D insufficiency. We propose that African
Americans with BMI  30 kg/m² should be specifically screened for vitamin D deficiency.
          Research grant provided by the Office of Dietary Supplements, NIH, Bethesda, 20892

CLINICAL POSTER: Obesity (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                        93
                                                                                        Obesity
P3-467 News Summary
Obese African Americans, especially women, have a high risk of vitamin D deficiency

Increased body weight and dark skin pigmentation are important risk factors for vitamin D
deficiency, according to a study being presented on Saturday, June 21, at The Endocrine
Society’s 85th Annual Meeting in Philadelphia. Obese African Americans, especially obese
African American women, are at highest risk, with an additional risk for hyperparathyroidism,
when one or more parathyroid glands become overactive. For these reasons, researchers
recommend that obese African Americans, especially women, should be routinely screened for
vitamin D deficiency.

Dark pigmentation of the skin as well as increased body weight have been previously described
as risk factors for vitamin D deficiency. Therefore, Dr. Shamik Parikh and colleagues at the
National Institute of Child Health and Development and other branches of the National Institutes
of Health in Bethesda, Md., examined the levels of vitamin D in a group of overweight African
Americans and Caucasians. They confirmed this earlier finding and also found that around 70
percent of obese African Americans, especially obese African American women, had a very high
prevalence of low vitamin D levels. In addition, most of these patients also had a secondary
hyperparathyroidism, which leads to too much calcium in the blood, apparently as a result of the
low vitamin D levels. Left untreated, hyperparathyroidism can cause additional health problems,
such as kidney damage or kidney stones and a loss of calcium from bones, leading to
osteoporosis.

The researhers say that they still do not fully understand the consequences of having low vitamin
D levels on body health, although a lot of research has been done with respect to vitamin D
levels and bone health. However, it is clear, they add, that the suboptimal levels of vitamin D in
the body are responsible for lower dietary calcium absorption from the stomach as well as
secondary hyperparathyroidism, which increases parathyroid hormone levels in an effort to
maintain normal calcium levels. Appropriate screening of the high-risk group of obese African
Americans can help detect people who can benefit from dietary modification, including vitamin
D supplements.

This study was supported the Office of Dietary Supplements of the National Institutes of Health.

                                              # # #




                                                94
                                                                                                          Obesity
P1-267
High-Throughput Phenotypic Analysis of Mice Identifies Novel Obesity- and Diabetes-Related Drug Targets.
David R Powell*1, Robert Brommage1, Gregory K Fontenot1, Urvi J Desai1, Christopher M DaCosta1, Deon D
Smith1, Kristal A Casias1, Jason P Gay1, Dorit B Donoviel1. 1Endocrinology, Lexicon Genet Inc, The Woodlands,
TX.

Screening gene function in mice in vivo is a powerful way to discover novel drug targets in the human genome (1).
To identify obesity- and diabetes-related drug targets, we are generating and phenotyping ~20 mouse knockout (KO)
lines/week; mutated genes encode proteins that are secreted or in families that are known targets of small molecule
drugs. Homozygous null (hom) and wild-type control littermates from each line are screened at 4 months of age for
weight, glucose tolerance by intraperitoneal GTT, % body fat and lean body mass by DEXA, serum and urine
glucose, and serum insulin. In general, lines are considered for further analysis if hom values differ by at least 1.5
SD from values of historical control mice. To date, > 750 KO lines including > 3000 wild-type control mice have
been phenotyped. Perilipin and melanin-concentrating hormone 1 receptor KO lines (the only published lean lines
studied) and the melanocortin-3 receptor KO line (the only published obese line studied) were all clearly identified
by this screen. Novel diabetes phenotypes were found in mice lacking either calcium channel alpha 2/delta 1 subunit
(CACNA2D1) or insulin degrading enzyme (IDE). CACNA2D1-deficient mice had glycosuria, impaired glucose
tolerance and low serum insulin, with further studies showing improved insulin tolerance and decreased insulin
immunoreactivity in pancreatic islets; calcium channel alpha 1D subunit-deficient mice have a similar phenotype
(2). Male IDE-deficient mice were glucose intolerant with high serum insulin. Importantly, the initial screen and
later confirmatory studies also found obesity and high serum insulin in mice lacking a G-protein coupled receptor,
and found low body fat with improved glucose tolerance and normal weight in 2 additional KO lines; in all 3 lines,
the gene mutated is a potential drug target not previously linked to obesity or diabetes. In 22 additional KO lines
where hom mice were of normal weight, the screen found low body fat (N=8), improved glucose tolerance (N=9), or
both (N=5); each line is being analyzed further. In summary, this high-throughput screen of KO mouse lines clearly
identifies both known and novel obesity- and diabetes-related phenotypes. We suggest that the novel phenotypes
result from inactivation of genes that are high quality targets for new drugs designed to treat human obesity and
diabetes.
          Reference: 1. Zambrowicz & Sands; Nat Rev Drug Discov 1:38, 2003
          2. Namkung et al, JCI 108:1015, 2001

BASIC POSTER: Obesity (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                         95
                                                                                         Obesity
P1-267 News Summary
Early research identifies potential drug targets for treating obesity and diabetes

New research in mice has identified known and novel genes with clinical characteristics that
make them potential drug targets for lowering the excess body fat and/or improving the insulin
resistance present in obese and diabetic patients, according to a new study being presented on
Thursday, June 19, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

The worldwide epidemics of obesity and type-2 diabetes have spurred a search for new drugs to
treat these conditions. Most drugs work by changing the action of a specific protein. Because
fewer than 100 proteins are the targets for all prescription drugs marketed, new approaches are
needed to identify the rare proteins with value as drug targets for obesity and diabetes.

Researchers at Lexicon Genetics have taken the approach to selectively turn off (knockout or
KO) individual genes in mice to block manufacture of the protein product of that gene. To
rapidly identify rare and novel obesity- and diabetes-related drug targets, the company has
industrialized the process so that they now make and clinically screen approximately 20 mouse
KO lines a week; they knock out only genes that are members of known drug target families and
they clinically screen young adult mice with tests routinely used to study humans, including
weight, DEXA to analyze body fat and lean body mass, glucose tolerance tests, serum and urine
glucose levels and serum insulin levels. To date, they have made and screened more than 750
KO lines.

They found the following: low body fat in the only two published lean lines screened (perilipin
and melanin concentrating hormone 1 receptor KO lines); high body fat in the only two
published obese lines screened (melanocortin-3 receptor and bombesin receptor subtype-3 KO
lines); signs of impaired insulin action in the only published insulin resistant line screened
(insulin degrading enzyme KO line) and insulin deficiency in mice lacking the calcium channel
alpha 2/delta 1 subunit, similar to the insulin deficient state reported in KO mice lacking the
related calcium channel alpha 1D subunit gene.

These findings show that the clinical screen used identifies genes relevant to obesity and diabetes
drug discovery. The clinical screen and later confirmatory studies also found a two- to three-fold
increase in body fat along with high insulin levels, suggesting a pre-diabetic state in young mice
lacking a specific G-protein coupled receptor. They further found low body fat with improved
glucose tolerance in two additional KO lines. In all three of these KO lines, the inhibited gene is
a potential drug target not previously linked to obesity or diabetes. In a number of additional
lines, the screen found low body fat, improved glucose tolerance, or both; each line is being
analyzed further as a potential obesity/diabetes drug target.

This research is supported by Lexicon Genetics of The Woodlands, Tex.

                                              # # #




                                                96
                                                                                                          Obesity
P3-458
Pharmacological Treatment of Hyperinsulinemia Obesity Syndrome in Children.
Fuad Ziai*1, Hanna S Sahhar1. 1Peds, Christ Hosp Med Ctr, Oak Lawn, IL.

We have treated children with significant hyperinsulinemia and obesity with combination drug therapy including
Amphetamine multi-compounds (Adderall) and Metformin (Glucophage XR). Thirty-three patients completed 12
months of treatment at the time of this report. They were divided into three age groups of <11 yrs, 11-14 yrs, and
>14 yrs. There were 15 males and 18 females, 21 Caucasians, 6 Hispanics, 1 African American, and 5 others. One
subject in the first group, three in the second group, and two in the third group failed to improve and were excluded
from the analysis.
The mean Body Mass Index (BMI) was 30.7, 36.2, and 41.8 at the baseline which decreased to 27.7 (p=0.004), 32.2
(p=0.001), and 37.4 (p=0.000) respectively by the 12th month of treatment. Blood glucose levels were normal in all
subjects both at fasting and two hour post prandial. The baseline mean insulin levels however, were elevated at 35.2
and 106.8 IU/ml at fasting and two hour post prandial which decreased to 15.4 (p=0.005) and 44.5 IU/ml (p=0.031)
by 12 months of therapy.
Conclusion: Combination drug therapy, Amphetamine compounds and Metformin can induce significant sustained
weight loss and improve insulin resistance.
    Comparison of the mean BMI in the age
                       groups
TIME                     BMI
                         <11yr (n=8)               11-14yr (n=8) >14yr (n=11) Total (n=27)
Initial                  30.71                     36.21          41.8           36.86
3 months                 29.55                     35.21          40.65          35.75
6 months                 28.4                      33.14          38.42          33.89
9 months                 28.11                     33.68          38.11          33.83
12 months                27.78                     32.24          37.48          33.05
There is a significant difference of BMI between the groups (Bonferroni & Tukey Test, p,0.005).

                  Comparison of the fasting and 2 hour post prandial insulin pre and post treatment
                                  Std.            Std. Error of       95% Confidence                   Sig. (2-
                           Mean                                                                 t   df
                                  Deviation       Mean                Interval                         tailed)
Pair 1 BASEFA-
                            19.87 26.35            6.21               6.76-32.97                3.2 17 0.005
POSTFAST
Pair 2 BASE2HR-
                            62.3 112.42            26.5               6.40-118.21               2.35 17 0.031
POST2HR
Paired-sample T-Test is used to compare the fasting insulin and 2-hr insulin separately. There are significant
differences between pre and post-test (fasting: p=0.005; 2-hr: p=0.031).

CLINICAL POSTER: Obesity (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                         97
                                                                                         Obesity
P3-458 News Summary
Weight-loss and diabetic drugs together helped children lose weight, reduce diabetes risk

Overweight children who took the combination of weight-loss and diabetic drugs lost a
substantial amount of weight and their insulin levels returned to near normal, according to a new
study being presented on Saturday, June 21, at The Endocrine Society’s 85th Annual Meeting in
Philadelphia

Childhood obesity has been a growing problem among United States population. The
consequences of being overweight can include diabetes, heart disease and hypertension in later
life. By far, there has been no effective method to treat this condition in childhood and
adolescence. Lifestyle modification – including improving nutrition and decreasing calorie
consumption while increasing physical activities – is successful but often is not maintained.

The pharmacological approaches currently used in adulthood obesity suffer from similar
limitations. To overcome current barriers, Drs. Fuad Ziai and Hanna S. Sahhar of Christ
Hospital Medical Center in Oak Lawn, Ill., began testing two well-known medications for a new
approach. The first, amphetamine compounds – through their effect on the hypothalamus and
the satiety center – have been used for appetite suppression and weight loss. Such a compound
was introduced several years ago to manage attention deficit disorder and attention deficit
hyperactivity disorder, with proven safety records in children. The second, metformin, is a
known medication for the management of type-2 diabetes in adolescents and adults.

In the current study, the researchers used the combination of these two drugs in children and
adolescents to address the problem of hyperphagia – an abnormally increased appetite for food
frequently associated with injury to the hypothalamus – and remedy hyperinsulinemia, an excess
of insulin in the blood. They started by choosing patients who had been referred to their clinic
for the management of obesity and had undergone a metabolic workup, which included
evaluating their glucose and insulin levels before and two hours after a meal.

The first 33 patients who completed a year of treatment had significantly high levels of insulin
with normal blood glucose, indicating a status of pre-diabetes. The range in age was 5–18 years
old, with approximately half from each sex and the majority being Caucasians, followed by
Hispanics and one African American. The majority of these children responded remarkably well
to treatment, especially during the first six months of therapy, with sustained weight control by
the 12th month. The insulin levels were abnormally high in the beginning but decreased
substantially towards normal ranges.

These results show that pharmacological management using both amphetamine compounds and
metformin is effective in controlling obesity and hyperinsulinemia in children and adolescents
and may be considered as an option to address this growing public health issue. The great
majority of patients tolerated these medications very well, with very few transient side effects,
including insomnia, irritability, emotional problems and gastrointestinal disturbances.
                                               # # #


                                                98
                                                                                                        Obesity
P3-457
Sibutramine / Orlistat Combination Offers No Advantage over Single Agent Therapy in Obese Women.
Galina Shenkerman*1, Etty Osher1, Naftali Stern1. 1Endocrinology, Metab and Hypertens, Tel Aviv Sourasky Med
Ctr, Tel Aviv, Israel.

Although significant weight loss can be attained by either sibutramine (SIB) or orlistat (ORL), the achieved weight
loss still falls short of target range in nearly all treated subjects. Because SIB and ORL operate via unrelated
mechanisms we postulated that combination therapy might have additive or even synergistic effects on weight. We
designed an open-label pilot study comparing the effects of SIB (15 mg/day, ORL (360 mg/day) or their
combination for 24 weeks in 50 women with a mean weight of 101.9±2.57kg and body mass index (BMI)
38.62±0.78Kg/m2. Patients were divided randomly into three groups to receive ORL (n=18); SIB (n=16,); or
SIB+ORL (COMB; n=16). At randomization there was no significant among-group difference in mean weight
(ORL group-103.87±5.0 kg, SIB group- 95.61±3.27kg and COMB group 105.77±4.28kg, p= NS) and BMI (ORL
group-40.3±1.55kg/m2, SIB group- 36.05±1.0 kg/m2, COMB group- 39.42 ±0.99kg/m2, p= NS). However the
initial waist as well as waist to hip ratio (WHR) were lower in the SIB group than in the two other treatment group
(waist: SIB - 104.4± 2.72cm, ORL -115.9±3.5cm, COMB -114.5±2.84cm, p=0.014; WHR: SIB- 0.84;ORL -0.93,
COMB - 0.91;p= 0.04). Withdrawal rates from the study were: ORL - 2/18, SIB-4/16, COMB-4/16 patients. After 6
months of treatment all groups showed significant (p<0.001) but entirely indistinguishable weight loss: [ORL-
6.1±1.13 kg (5.9%), SIB - 8.36±1.65kg (9.2%), COMB - 8.17±2.0kg (8.0%)] along with similar decrease in BMI
[ORL -6.2 %, SIB- 9.8%, COMB-8.13%; p=ns] and waist circumference [ORL group-5.17%,SIB group-
8.4%,COMB group-6.55%;p=ns]. All treatment groups experienced significant and essentially comparable
improvement in metabolic profile (CHOL: -6.0%, -6.1%, -18,3%; TG -29.7%, -18.7%, -21.2%; GLU -10.9%, -
4.7%, -15.5% in ORL, SIB, COMB, respectively) and a similar decline in blood pressure (ORL: –11 ±4 / –6.7
±2.2mmHg , SIB: –5.7 ±3.7 / –2.2 ±2.9mmHg , COMB : –8.9 ±4.2 /–8.3 ±3.3mmHg for systolic / diastolic pressure,
respectively; p= ns) . Conclusion: While SIB and ORL exerted significant and comparable effects on weight there
was no benefit in combining ORL/SIB. Whether longer treatment can elicit such responses or this outcome simply
reflects endogenous counter-mechanisms limiting the degree of attainable weight loss remains subject to further
studies.

CLINICAL POSTER: Obesity (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                        99
                                                                                       Obesity
P3-457 News Summary
Combining the weight-loss power of two drugs does not add up

Combined therapy with two effective weight-loss drugs is not more effective in inducing weight
loss than either drug alone, according to a new study being presented on Saturday, June 21, at
The Endocrine Society’s 85th Annual Meeting in Philadelphia.

Two drugs, sibutramine or orlistat, are currently approved for weight loss. Even though the two
drugs have been used for several years now, the usefulness of combined therapy had not been
evaluated.

Therefore, Drs. Galina Shenkerman, Etty Osher and Naftali Stern of the Tel Aviv Sourasky
Medical Center in Tel Aviv, Israel designed a controlled and prospective study to determine, for
the first, time, their combined benefit. They studied 50 obese women with an average age of 48,
who received sibutramine, orlistat or both drugs for six months. Each of the treatment protocols
resulted in significant reduction in weight, waist circumference, waist to hip ratio and
cholesterol. However, there were no differences in outcome among the three treatment groups.

                                             # # #




                                              100
                                                                          Obesity: Control of Appetite
OR33-2
Meal-Stimulated Release of the Putatitive Satiety Hormone PYY in Severe Obesity and Following Gastric
Bypass Surgery.
Carel W le Roux1, Simon JB Aylwin*2, Frances Coyle2, Mohammed Ghatei1, Ameet Patel3, Steven R Bloom1.
1
  Metab Med, Imperial Coll at Hammersmith campus, London, United Kingdom; 2Endocrinology, King’s Coll Hosp,
London, United Kingdom; 3Surg, King’s Coll Hosp, London, United Kingdom.

Background
The gut hormone PYY is expressed in the ileum and released in response to the ingestion of food. PYY has recently
been proposed as a satiety factor, acting centrally to inhibit neuropeptide Y (NPY) release from the arcuate nucleus
by binding specifically to the inhibitory type 2 NPY receptor (Y2R). Surgical treatment of obesity has an anorectic
effect and can lead to marked weight loss. We investigated the PYY response to a meal in morbidly obese subjects
and in patients who had previously undergone surgical procerdures for obesity.
Methods
The PYY response to a standard meal was evaluated in female patients who had previously undergone either gastric
bypass surgery (n = 5; post-procedure BMI: 36.8kg/m²) or vertical banded gastroplasty (n = 7; post-procedure BMI
36.3kg/m²); and compared to patients with severe obesity matched to pre-procedure BMI (n =10; BMI: 46.3kg/m²).
Following an overnight fast, plasma samples were collected before and at 30 minute intervals for 180 minutes
following a mixed 560kcal meal. Samples were stored at -20C and assayed for PYY.
Results
In a previous protocol basal PYY levels were documented at 14.1 pmol/L with a 47% increase after a standard meal.
We demonstrated that control obese patients had an attenuated and non-significant PYY response to 560kcal meal
(basal: 13.2, peak: 15.8pmol/L). Patients who had undergone gastric bypass, however, had an exaggerated PYY
response (basal: 13.8, peak: 36.3pmol/L; p<0.001 by ANOVA). This exaggerated response was not seen in patients
who had undergone vertical banded gastroplasty (basal 11.3, peak 14.83pmol/L). The response amongst patients
following gastric bypass was significantly greater than amongst obese controls (p<0.001) or patients following
gastroplasty (p<0.001).
Conclusions
Morbidly obese subjects have an attenuated release of the putatitive satiety factor PYY. Following gastric bypass
surgery performed for morbid obesity, release of PYY is exaggerated. This mechanism is consistent with the
reduced appetite observed in these subjects and may contribute to their weight loss.




CLINICAL ORAL: Obesity: Control of Appetite (1:00 PM - 2:30 PM)

Presentation Date: 6/21/2003, Time: 1:15 PM; Location: Lecture Hall




                                                        101
                                                               Obesity: Control of Appetite
OR33-2 News Summary
Newly found hormone released by stomach may serve as basis for future obesity therapy

A newly discovered hormone that is released by the stomach after a meal, or a drug like it, could
be a future treatment for people who are seriously overweight, according to a new study being
presented on Saturday, June 21, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

Severe obesity is increasingly seen as a disorder of appetite – a failure to recognize an adequate
energy store or an appropriate meal. Recently, research has demonstrated that genetic disorders
of appetite control are the cause of some of the more extreme forms of obesity. Ultimately, an
area of the brain called the hypothalamus acts as the sensor for food stores and recent meal size
and drives the opposing sensations of hunger and fullness. The release of certain hormones
(chemical messengers) from fat deposits and the stomach accounts for one mechanism for the
transmission of these signals to the brain, and the regulation of appetite and feeding.

Surgical procedures that reduce stomach size – gastric banding – or divert food away from the
stomach and duodenum –gastric bypass – are very effective and established treatments for severe
obesity. They seem to cause a decrease in appetite among patients and an increased ability to
accept a low-calorie diet. The reduction in appetite and dramatic weight loss are unexplained,
which led to a collaborative research project between the King’s College Hospital and Imperial
College at the Hammersmith Hospital in the United Kingdom. Because surgery for obesity is
becoming more popular, the researchers believe that it is necessary to understand the
mechanisms that are involved if the procedures are to be improved.

This research questioned whether the observed appetite reduction and weight loss might be
related to an increased release of a hormone called PYY. PYY seems to be one of the chemical
signals to the brain to stop a person from continuing to eat. The researchers measured the PYY
hormone blood level before and after a 560-calorie meal in female patients who had previously
undergone either gastric bypass surgery (5 patients) or vertical-banded gastroplasty (7 patients).
These PYY levels were compared to those from patients with severe obesity, matched to the
same weight for height as the patients had before their operations.

Patients who underwent gastric bypass had very high levels of the PYY hormone after a meal,
leading the researchers to believe that this hormone surge is responsible for the loss of desire to
eat excessively. In addition, PYY hormone levels in gastric bypass patients were significantly
greater than among obese controls or patients following gastroplasty. Interestingly, compared to
results from a previous study in normal weight volunteers, this study found that control obese
patients had a slightly lower response than lean people. If these results are confirmed by future
studies, the researchers note that this may mean that the PYY hormone – or a drug like it – could
be used to treat people who are seriously overweight.

This research was supported by Welcome Trust and King’s College Hospital Research and
Development Department.
                                          # # #


                                               102
                                                                           Obesity: Control of Appetite
OR33-1
Impact of Sleep Curtailment on Sympathovagal Balance, Leptin Levels, Hunger and Appetite.
Plamen Penev*1, Leonard Hudson1, Karine Spiegel1,2, Esra Tasali1, Rachel Leproult1,2, Eve Van Cauter1. 1Sect of
Endocrinology, Dept of Med, Univ of Chicago, Chicago, IL; 2Ctr d’Etude des Rythmes Biologiques, Lab de Physiol,
Univ Libre de Bruxelles, Brussels, Belgium.

Sympathetic activation is a known inhibitor of leptin release. Since sleep deprivation can be associated with
increased sympathetic activity, we examined the effect of bedtime restriction on cardiac sympathovagal balance,
leptin levels, hunger and appetite.
Twelve healthy subjects (age 22 ± 1y, BMI 23.6 ± 0.6 kg/m²) were studied in the laboratory on 2 occasions. Each of
them spent 2 consecutive nights with 4-h bedtimes followed by a night with 8 hrs in bed during one of the studies,
and 2 nights with 10-h bedtimes followed by a night with 8 hrs in bed during the other. After the 2 nd night of sleep
the subjects remained at bed rest until the end of the study. A constant glucose infusion (5g/kg/24h) was started at
8:00 and blood samples for leptin measurement (RIA, Linco Research) were obtained every 20 min until 21:00.
Subjects completed visual analog scales for hunger and appetite every hour from 9:00 to 21:00. Heart rate and beat-
to-beat intervals were recorded continuously (Mini-Logger, Mini-Mitter). The autocorrelation coefficient of
consecutive interbeat intervals (rRR), an index of sympathovagal balance, was calculated every 5 min for the last 36
hrs of each study.
Total sleep time was 9h 8min ± 11min with 10-h bedtimes and 3h 53min ± 2min with 4-h bedtimes (p=0.005).
Despite identical levels of caloric intake and energy expenditure, the 4-h bedtime condition was associated with a
19% decrease in leptin (p=0.041), a 16% increase in hunger (p=0.019), and a 16% increase in appetite (p=0.023),
particularly for high fat and high carbohydrate foods (+24%, p=0.034). Overall, subjects had faster heart rates in the
4-h bedtime condition, particularly in the early morning (60 ± 1 vs. 55 ± 1 bpm, p=0.032), as well as during the
night with 8-h bedtimes (55 ± 1 vs. 52 ± 1 bpm, p=0.048). Sleep curtailment also resulted in increased rRR both at
night (0.50 ± 0.02 vs. 0.44 ± 0.02, p=0.037) and for 12 hrs after awakening (0.62 ± 0.01 vs. 0.57 ± 0.01, p=0.021).
The higher rRR values and the increase in heart rate in the restricted sleep condition indicate a shift in cardiac
sympathovagal balance in favor of the sympathetic component.
We conclude that sleep restriction alters leptin signaling of caloric need and stimulates hunger and appetite.
Increased sympathetic activity may be involved in mediating the effects of sleep loss on the endocrine control of
energy balance and the cardiovascular system.

CLINICAL ORAL: Obesity: Control of Appetite (1:00 PM - 2:30 PM)

Presentation Date: 6/21/2003, Time: 1:00 PM; Location: Lecture Hall




                                                         103
                                                                 Obesity: Control of Appetite
OR33-1 News Summary
Sleep restriction is linked to weight gain and risk for obesity

Sleep restriction can stimulate hunger and appetite and alter the hormonal signaling of caloric
need, according to a study being presented on Saturday, June 21, at The Endocrine Society’s 85th
Annual Meeting in Philadelphia. These results raise the possibility that a decrease in levels of
the hormone leptin triggered by chronic sleep restriction, despite adequate food supply and a
sedentary lifestyle, may increase susceptibility for weight gain and the long-term risk for obesity.

Acute sleep deprivation results in increased activity of the sympathetic nervous system – a part
of the nervous system that plays an important part in the response and survival when exposed to
adverse events and stressful stimuli or illness. More prolonged sleep loss in rodents is associated
with significant alterations in energy balance and increased food intake. Leptin – found in fat
cells – is one of the main hormones that signal changes in energy balance to the brain and help
match our food intake and caloric expenditure. A reduction of body fat stores leads to a decrease
in leptin levels, whereas excess body fat is associated with increased leptin.

Because the activation of the sympathetic nervous system is a known inhibitor of leptin release,
Dr. Plamen Penev and colleagues in Chicago and Belgium wanted to examine the effect of
recurrent partial bedtime restriction on sympathetic activity, leptin levels, hunger and appetite.
Twelve healthy people, with an average age of 22 years old and body mass index of 23.6 ± 0.6
kg/m2, completed two studies in random order at least six weeks apart. In the sleep laboratory,
each subject spent two consecutive nights with four-hour bedtimes, followed by one night with
eight hours in bed during one of the studies, and two nights with 10-hour bedtimes, followed by
one night with eight hours in bed during the other. After the second night of sleep in both
studies, the participants remained at bed rest and received a constant glucose infusion instead of
meals to maintain the same level of caloric intake and energy expenditure. They rated their
hunger and appetite at hourly intervals. Leptin and heart rate variability were monitored.

Despite identical levels of caloric intake and energy expenditure, the four-hour bedtime
condition was associated with a 19-percent decrease in daytime leptin levels, a 16-percent
increase in hunger and a 16-percent increase in appetite, particularly for high fat and high
carbohydrate foods, a 24 percent increase. The participants had a significantly higher overnight
heart rate with restricted sleep, which remained elevated even when the participants were
exposed to identical eight-hour bedtimes. Sleep curtailment was also associated with
deterioration in overnight heart rate variability, which was particularly pronounced in the early
morning. The changes in heart rate and variability with restricted sleep indicate a shift in the
neural control of the heart towards a more “stress-like” state of sympathetic predominance. The
changes in heart rate and its variability in the restricted sleep condition suggest that increased
sympathetic activity may play an important role in the mechanism of these hormonal changes.

This study was supported by the National Institutes of Health.

                                              # # #


                                                104
                                       Central Nervous System & Gut Regulatory Peptides
P3-93
Dissociation of Ghrelin Action on GH-Stimulation and Weight Gain.
Michael D Culler*1, Jesse Z Dong1, Yeelana Shen1, Jundong Zhang1, Daniel De Oliveira1, John E Taylor1, Rakesh
Datta1. 1Biomeasure, Inc/Beaufour-IPSEN Grp, Milford, MA.

Ghrelin is a 28 amino acid peptide that was recently identified as an endogenous ligand for the growth hormone
(GH) secretagogue (GHS) receptor. Ghrelin has been demonstrated not only to stimulate GH secretion, but also to
increase food intake and body weight gain. We recently identified an analog of human (h) ghrelin, BIM-28163, that
is an antagonist at the h-GHS receptor. In vivo, BIM-28163 can completely abolish the ability of h-ghrelin to
stimulate GH secretion. In the present study, we have examined the effects of BIM-28163 on weight gain in normal,
adult rats. Male, Sprague Dawley rats (250g) were housed in individual cages and maintained under 12:12 hour
light:dark conditions with both food and water available ad libitum. The rats were injected 3x/day (0800, 1200, and
1600h), ip, with either BIM-28163 at 40 or 400nmoles/kg or vehicle for 7 days. For comparison, another group of
rats was injected with 40nmole/kg BIM-28167, another h-ghrelin analog with comparable in vitro plasma half-life as
BIM-28163. Unlike BIM-28163, BIM-28167 is a full agonist at the h-GHS receptor and stimulates GH-secretion in
vivo. Individual body weights and food consumption were measured daily. By day 7, the 400nmole/kg treatment
with BIM-28163 had stimulated a significant (p<0.008) 14.9 ± 4.3g body weight increase as compared with the
vehicle-injected control animals. The 40nmole/kg BIM-28163 treatment was without effect. In comparison,
treatment with the h-GHS receptor agonist, BIM-23167, also stimulated a significant (p<0.03) 9.3 ± 2.4g increase in
body weight as compared with controls. Cumulative food intake over the 7-day treatment period was similarly,
though non-significantly, increased by treatment with 400nmole/kg BIM-28163 and 40nmole/kg BIM-28167 (14.7 ±
1.7g and 15.9 ± 1.5g over food intake by vehicle-injected controls, respectively). The 40nmole/kg treatment with
BIM-23163 was without effect on food intake. The results of this study demonstrate that BIM-23163, which acts as
an antagonist at the h-GHS receptor, and which blocks ghrelin-stimulated GH secretion in vivo, acts as an agonist
with regard to the ghrelin property of stimulating increased weight gain. These divergent results suggest that ghrelin
influences weight gain through a receptor other than the presently known GHS receptor. The present observations
demonstrate the feasibility of creating ghrelin analogs that are selective for specific ghrelin activities.

BASIC POSTER: Central Nervous System & Gut Regulatory Peptides (11:00 AM - 12:00 PM and 2:30 PM - 3:30
PM)

Presentation Date: Saturday, June 21, 2003




                                                        105
                                  Central Nervous System & Gut Regulatory Peptides
P3-93 News Summary
Early study shows promise for creating weight-loss drugs based on hormone

Researchers have created a drug based on a weight-regulating hormone that can be used to
selectively stimulate or inhibit weight gain, according to early research being presented on
Saturday, June 21, at The Endocrine Society’s 85th Annual Meeting in Philadelphia

More than 60 percent of U.S. adults and a considerable percentage of children are considered
overweight or obese. Recent studies have implicated a new hormone, named ghrelin, as a
regulator of appetite and weight gain, and changes in ghrelin levels have been associated with
both successful and failed weight-loss efforts. It has also recently been publicized that ghrelin
levels remain low in patients who have lost weight by undergoing gastric bypass surgery, most
likely partially explaining the lack of appetite in these individuals.

The opposite, but equally serious, problem of lack of appetite and weight gain in the elderly and
the chronically ill may also potentially be corrected by ghrelin-based drugs. These results have
fostered the idea that drugs that can alter ghrelin action might be useful in controlling appetite
and body weight. The problem is that ghrelin is known to have a number of other actions in the
body; therefore, altering ghrelin action could produce undesirable effects. One such action of
ghrelin is alteration of growth hormone (GH) secretion.

To overcome this problem, researchers at Biomeasure, Inc. in Boston have invented a unique,
modified version of ghrelin, named BIM-28163, that can alter weight gain, while preventing
ghrelin-induced changes in growth hormone secretion. They have also developed other ghrelin-
based drugs, none of which has yet proceeded to clinical tests.

In the current study, Dr. Michael D. Culler and colleagues examined the effects of BIM-28163
on weight gain in normal, adult rats. The rats were injected three times a day for seven days,
with either 40 or 400nmoles/kg or a placebo. For comparison, another group of rats was injected
with 40nmole/kg of BIM-28167, another ghrelin-based drug but one that stimulates GH-
secretion in vivo. Individual body weights and food consumption were measured daily. By day
seven, the 400nmole/kg treatment with BIM-28163 had stimulated a significant body weight
increase as compared with the placebo-injected control animals. The 40nmole/kg BIM-28163
treatment had no effect. In comparison, treatment with BIM-28167 also stimulated a significant
increase in body weight as compared with controls. Food intake over the seven-day treatment
period was similarly, though non-significantly, increased by treatment with 400nmole/kg BIM-
28163 and 40nmole/kg BIM-28167.

These results show that it is feasible to create drugs that can selectively alter specific actions of
ghrelin, such as controlling weight gain without affecting other ghrelin actions in the body.

This study was supported by Biomeasure, Inc., the U.S. research division of Beaufour-IPSEN.

                                                # # #


                                                 106
                         Central Nervous System, Gut Regulatory Peptides & Obesity
OR36-1
Melanin-Concentrating Hormone (MCH) Ablation in ob/ob Mice Attenuates Phenotypic Manifestations of
Leptin Deficiency and Increases Energy Expenditure.
Gabriella Segal-Lieberman*1, Richard H Bradley1, Efi Kokkotou1, Xiaomei Wang2, Daniel Trombly1, Michael
Carlson1, Sarah H Bates1, Martin G Myers, Jr1, Jeffrey S Flier2, Eleftheria Maratos-Flier1. 1Obesity Sect/Res Div,
Joslin Diabetes Ctr, Boston, MA; 2Div of Endocrinolgy, Beth Israel Deaconess Med Ctr, Boston, MA.

Leptin deficiency underlies the phenotype of the ob/ob mouse, which is characterized by multiple abnormalities
including hyperphagia, marked reductions in energy expenditure and glucose intolerance. MCH is a hypothalamic
peptide important in mediating energy homeostasis. ICV administration of MCH increases appetite and mice lacking
the MCH gene are lean. As MCH is overexpressed in ob/ob mice we predicted that ablation of MCH in these leptin
deficient mice would lead to attenuation of the ob phenotype. MCH null mice backcrossed 7 generations onto a
C57BL/6 background were crossed with ob/ob mice on the same background yielding mice lacking both MCH and
leptin (double null). Compared to ob/ob mice, double null mice showed a marked reduction in weight (67±1.8 vs
54.5 ± 2.3 grams) secondary to decreased total body fat (37.5±2.2 vs 24.3±.08 grams of fat). Lean body mass was
increased by 1 gram. Decreased obesity was associated with improved glucose tolerance, however insulin levels
remained high. Corticosterone levels were signficantly decreased in double null mice compared to ob/ob mice,
although they were still mildly elevated compared to control animals. Surprisingly, double null mice were as
hyperphagic as ob/ob mice. Improvement of body weight was associated with a 3-fold increase in locomotor activity
as well as an increase in resting energy expenditure. Compared to ob/ob, double null mice had increased core
temperature, markedly improved cold tolerance and two fold increased UCP-1 protein levels in brown adipose
tissue. Expression of stearoyl-CoA desaturase 1, which is increased in ob/ob animals and is implicated in mediating
the effects of leptin deficiency, was significantly decreased in double null animals although still elevated compared
to control animals (ctl=83±20, ob/ob=373±58, double null=232±36 p=.02, arbitrary units) . Furthermore, double null
animals had normal bone length and partial normalization of brain weight, in contrast to the growth deficits seen in
ob/ob mice.
This study defines a novel, critical role for MCH in maintaining energy homeostasis through effects on both
locomotor activity and thermoregulation. While MCH is involved in the normal regulation of feeding it is not
necessary for the hyperphagia of leptin deficiency. In addition MCH appears to have a key role as a downstream
mediator of leptin action and is required for the full manifestation of leptin deficiency.

BASIC ORAL: Central Nervous System, Gut Regulatory Peptides & Obesity (11:15 AM - 12:45 PM)

Presentation Date: 6/22/2003, Time: 11:15 AM; Location: 113 B




                                                        107
                     Central Nervous System, Gut Regulatory Peptides & Obesity

OR36-1 News Summary
Brain peptides affect activity level and metabolism, leading to decreased weight

Activity and energy expenditure are regulated processes, and body weight can be altered by
regulating brain peptides that alter activity, according to a new study being presented on Sunday,
June 22, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

Obesity in both mice and humans results from an imbalance of eating and energy expenditure. It
is well known that appetite is in part mediated by factors like fat, the digestive system and the
brain, and an imbalance of these factors leads to overeating. The process by which energy
expenditure is regulated is not as well understood.

Dr. Eleftheria Maratos-Flier and colleagues in Boston studied obese mice (ob/ob) that lack leptin
– a peptide hormone produced by fat cells that suppresses appetite and burns fat. Their data
indicate that obesity can be reduced by taking away the brain peptide, melanin concentrating
hormone (MCH), which was initially identified to enhance appetite. However, ob/ob mice
missing MCH are leaner not because they eat less but because they are more active and have
higher metabolism than ob/ob mice that make MCH.

These results indicate that activity and energy expenditure are also regulated processes and that
body weight can be altered by regulating brain peptides that alter activity.

The study was supported by the National Institutes of Health.

                                              # # #




                                               108
                                                                          Prolactin Signaling & Action
P3-137
Prolactin Production and Release by Adipose Tissue from Morbidly Obese Patients.
Eric Hugo*1, Keith Gersin2, Adel Bakhsh2, Bonnie Neltner1, Nira Ben-Jonathan1. 1Cell Biol, Univ of Cincinnati,
Cincinnati, OH; 2Surg, Univ of Cincinnati, Cincinnati, OH.

The incidence of obesity in the US has reached epidemic proportion, with over 20% of adults classified as obese.
Adipose tissue consists primarily of terminally differentiated adipocytes with fewer preadipocytes capable of
proliferation and differentiation. Prolactin (PRL) is a pleiotropic hormone produced by both pituitary and
extrapituitary sites under dissimilar regulation. We previously reported de novo synthesis of PRL by breast adipose
explants. Here we have examined endogenous PRL in adipose tissue from obese patients (BMI: 47-80) undergoing
gastric bypass surgery. The specific aims were to: a) compare PRL release from visceral and subcutaneous explants,
b) correlate PRL and PRL receptor (PRL-R) expression during preadipocyte differentiation with early and late
markers of adipogenesis, and c) determine the effects of PRL immunoneutralization on preadipocyte differentiation
and endocrine function. Explants were cultured in serum-free medium, and media collected on days 1, 3, 7, and 10
were analyzed for PRL by a bioassay. PRL release was 5-20 fold higher in visceral than subcutaneous explants, with
some variability among patients. PRL release from visceral explants markedly increased between days 3 and 7
whereas that from subcutaneous explants was unchanged. Preadipocytes isolated from the two fat depots were
induced to differentiate and cultured for 14 days with either hPRL antisera or control serum. Expression of PRL,
PRL-R, lipoprotein lipase (LPL), PPAR, leptin, and adiponectin was examined by RT-PCR. Morphological
differentiation was assessed by lipid staining. PRL expression was unchanged during preadipocyte differentiation
whereas PRL-R expression increased progressively. PRL immunoneutralization resulted in increased LPL and
PPAR expression but reduced leptin and adiponectin, especially in visceral preadipocytes. In conclusion, this is the
first report on PRL expression and release by adipose tissue from obese patients. Reduction of secreted PRL by
immunoneutralization affects several key markers of adipogenesis and adipocyte-derived hormones, suggesting an
important role for PRL as a local adipogenic hormone.
          Supported by NIH grants ES10154 and CA80920.

BASIC POSTER: Prolactin Signaling & Action (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                        109
                                                                Prolactin Signaling & Action
P3-137 News Summary
First-time finding shows fat tissue in obese produces a hormone and possibly links it to fat
cell production

Researchers report for the first time that the hormone prolactin (PRL) is produced by body
cavity, or visceral, fat tissue and subcutaneous fat tissue in the obese, according to a new study
being presented on Saturday, June 21, at The Endocrine Society’s 85th Annual Meeting in
Philadelphia. The results also suggest that PRL may be important in fat cell production, a
finding which could lead to new ways of treating obesity.

The incidence of obesity in the United States has reached epidemic proportion, with more than
20 percent of adults classified as obese. Adipose tissue, or fat, consists of several different types
of cells, including mature fat cells that do not divide and precursor cells that can divide or
become mature fat cells. Identifying factors produced in fat that stimulate cell growth,
maturation or metabolism may provide targets to treat obesity.

Prolactin (PRL) is a hormone that stimulates milk production and was originally thought to be
produced only by the pituitary. It is now known that in addition to the pituitary PRL is produced
by other tissues, including white blood cells, prostate and breast, and has been shown to promote
cell growth and maturation.

In the current study, Dr. Eric R. Hugo and colleagues at the University of Cincinnati in
Cincinnati, Ohio, studied fat tissue from morbidly obese individuals undergoing stomach bypass
surgery. Small fragments of the tissue were maintained in culture medium and tested for PRL
release. PRL was found to be released into the medium after several days of incubation, with
more PRL produced by body cavity fat than subcutaneous fat. Isolated fat precursor cells from
these tissue samples were treated with drugs to stimulate their development into mature fat cells.

During the process of fat cell maturation, expression of the PRL gene and genes known to be
specifically active in fat cells, such as the hormone leptin gene, were monitored at the molecular
level. PRL was expressed in cells from both tissue sources during this process and the amount of
expression changed during fat cell maturation. Furthermore, removing PRL from the medium by
using an antibody that binds and inactivates PRL altered the expression of several fat-specific
genes during the process of maturation. This result suggests that PRL may be important in fat
cell development.

The researchers are currently conducting experiments to find out if PRL is involved in the
development and maintenance of fat tissue; if PRL has an effect on the metabolism of fat cells,
such as fat breakdown and production and if there are major differences in adipose-produced
PRL between obese and non-obese individuals. Answers to these questions, say the researchers,
could possibly lead to new methods for treating obesity.

                                               # # #



                                                110
                                                                                        Neuroendocrinology
OR2-3
Central Nervous and Metabolic Actions of Intranasally Applied Leptin.
Carla Schulz*1, Kerstin Paulus1, Hendrik Lehnert1. 1Dept of Endocrinology and Metab, Otto-von-Guericke Univ,
Magdeburg, Germany.

In obese people the ratio between CSF and serum leptin levels is diminished. This is assumed to be due to a
resistance of leptin-receptors, thus reducing the transport of leptin across the blood-brain-barrier (BBB). As a
consequence, peripherally elevated leptin levels can not adequately affect central nervous appetite regulation. This
resistance is also limiting a possible therapeutical use of leptin. A promising way to circumvent the BBB is the
intranasal administration of leptin via the vomeronasal organ.
In this study we present for the first time metabolic and central nervous peptide expression data in a study
employing the intranasal application of leptin. Male Wistar rats were intransally treated daily with leptin (0.1 or 0.2
mg/kg) or saline as control for 4 weeks at the beginning of the dark period. Body weight and food/water
consumption were measured daily (n=7 for leptin groups, n=10 for the control group). At the end of the observation
period animals were decapitated one hour after a final drug-application, trunc blood was collected and brains were
removed.
Metabolic parameters over the 4 week period were as follows (mean ± SD): Weight gain: 0.1 mg/kg: 79,4±6,3 g; 0.2
mg/kg: 80,8±12,3 g; control: 94,7±15,0 g. Food intake: 0.1 mg/kg: 588,7±22,8 g; 0.2 mg/kg: 616,1±30,8 g; control:
652,4±43,8 g. Water intake: 0.1 mg/kg: 710,3±34,7 g; 0.2 mg/kg: 756,5±55,6 g; control: 788,3±59,8 g. As assessed
by ANOVA for repeated measures, weight gain differed significantly between groups. Post-hoc analysis revealed
significant or highly significant differences between the leptin groups and the control group from day 7 until the end
of the experiment – weight gain did not differ between the leptin groups. Food intake differed highly significantly
between groups - post-hoc testing showed significant or highly significant differences starting at day 4. Water intake
differed significantly between groups.
Preliminary data for neuropeptide mRNA expression indicate a downregulation of NPY expression in the ARC and
an upregulation of CRF mRNA in the PVN of the hypothalamus in the leptin-groups, particularly at the higher dose.
Also elevated was CRF mRNA in the amygdala after leptin treatment.
From the results of our experiments we conclude that the intranasal application of leptin elicits central nervous
effects indicating a possible therapeutical use of this mode of administration. Data from in-situ hybridization in the
PVN confirm the CNS access of leptin.

BASIC ORAL: Neuroendocrinology (1:00 PM - 2:30 PM)

Presentation Date: 6/19/2003, Time: 1:30 PM; Location: 108 B




                                                         111
                                                                         Neuroendocrinology
OR2-3 News Summary
Hormone administered by nose drops reduced food intake, caused weight loss in animals

A hormone produced in fat cells reduced food intake, causing weight reduction, in animals that
received the treatment in nose drops, according to a study being presented on Thursday, June 19,
at The Endocrine Society’s 85th Annual Meeting in Philadelphia. These results provide
promising implications for further research and future developments of anti-obesity drugs.

Leptin is a hormone produced in body fat – also called adipose tissue – of all mammals,
including humans. The amount of leptin synthesized is proportional to the amount of fat.
Released into the circulation, it ends up in the brain’s hypothalamus, a major center of appetite
regulation. With its actions in the hypothalamus, it provides information to the brain on the
amount of body fat and influences food intake through a number of other substances, such as the
food intake stimulating neuropeptide Y (NPY) and the satiety producing corticotropin-releasing
factor (CRF). To reach the brain, leptin has to be shuffled by transporters across the blood brain
barrier, a protective structure that guards the brain against substances circulating in the blood.
However, it is known that in obese people leptin cannot reach the brain in adequate amounts
through the transporters, thereby misinforming the hypothalamus about the body fat mass. As a
consequence of this misinformation, food intake is inadequately high.

Dr. Carla Schulz and colleagues at Otto-von-Guericke University in Magdeburg, Germany,
developed an experimental design in rats in which a solution of leptin was administered in the
nasal cavity of the animals. There is no blood brain barrier preventing the olfactory nerve – the
nerve that transmits information on smell to the brain – from transporting leptin, which could
reduce appetite and, as a consequence, food intake by mimicking a large amount of body fat and
fooling the brain about energy stores.

They found in the experiments that animals treated with leptin in nose drops gained less weight
during the four weeks of treatment than the rats who received the drug-free solution, which
served as a control. Leptin-treated animals gained roughly 15 percent less weight than the rats
without hormone application. Statistical analysis revealed that the weight gain differed
significantly between leptin groups and the control group. Food and water intake differed
significantly between groups. The researchers believe the data show that the application of
leptin in nose drops is effective on food intake and body weight regulation and, thus, proof that
the intranasal application is an effective way to supply this hormone to the brain.

In another experiment, the researchers found in the hypothalamus an up-regulation of the satiety-
inducing CRF and a down-regulation of the food intake stimulating NPY in animals treated with
leptin. They believe that the changes of CRF mRNA in a region of the brain might be indicative
of an influence of leptin on emotional components of food intake.

                                              # # #




                                               112
Osteoporosis
                                                                                                Osteoporosis
P2-500
Sustained Effect of Risedronate Treatment over Seven Years in Postmenopausal Women with Osteoporosis.
O Sorensen1, S Goemaere2, D Wenderoth3, A Chines*3, C Roux4. 1Osteoporosis Res Ctr, Hvidorve Univ Hosp,
Hvidovre, Denmark; 2Unit for Osteoporosis and Metab Bone Dis, Ghent Univ Hosp, Ghent, Belgium; 3Procter &
Gamble Pharmaceuticals, Mason, OH; 4Dept of Rheumatol, Hosp Cochin, Paris, France.

In a multinational, double blind, placebo controlled study in women with established osteoporosis risedronate (RIS)
significantly decreased the incidence of vertebral fractures over 3 years. In a 2 year extension of this study, RIS
demonstrated a sustained antifracture effect. At the end of the 5 years, 164 women participated in a further 2 year
extension in which all patients were given RIS 5 mg daily. We now report the results for these women who either
continued another 2 years on RIS (7 yr RIS, n=83) or just initiated RIS (5 year PLBO/2 year RIS, n= 81).
Throughout the 7 years of the study, all patients received 1000 mg/d calcium and if baseline levels were low, up to
500 IU/day of vitamin D.
Over 7 years, lumbar spine (LS) BMD increased from baseline by 11.5% in the 7 yr RIS group and 6.1% in the 5 yr
PLBO/2 year RIS group. At the total hip, BMD increased by 3.9% and 1.0% in the respective groups. During years
6-7, increases in LS and total hip BMD were observed in both groups, with greater increases in the 5 yr PLBO/2
year RIS group, as expected. The annualized incidence of new vertebral fractures for this cohort is reported below.
Annualized Incidence of New Vertebral Fractures
               5yr PLBO/2yr RIS          7yr RIS
Years 0-3      7.6                       4.7
Years 4-5      12.3                      5.2
Years 6-7      3.8                       3.8
The cumulative incidence of osteoporosis-related nonvertebral fractures over 7 years was 25.9% and 15.7% in the 5
yr PLBO/2 yr RIS and 7 yr RIS groups, respectively. Overall adverse event profile was similar between the two
groups, including the incidence of upper gastrointestinal adverse events.
Treatment with risedronate over 7 years demonstrated continued BMD increases. The results of this extension study
strongly suggest sustained effects of risedronate on vertebral fractures.

CLINICAL POSTER: Osteoporosis (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                       113
                                                                                   Osteoporosis
P2-500 News Summary
Osteoporosis drug shows rapid and sustained ability to prevent fractures

Postmenopausal women with osteoporosis who took the drug risedronate (Actonel) for seven
years maintained bone strength – as exhibited by sustaining a reduced number of vertebral
fractures as in previous years taking the drug, according to a new study being presented on
Friday, June 20, at The Endocrine Society’s 85th Annual Meeting in Philadelphia. In addition,
those who recently began taking the drug showed a rapid increase in bone strength and few
fractures.

Risedronate is a non-hormonal drug used in the prevention and treatment of osteoporosis,
helping reverse bone loss and protect bones. Because osteoporosis treatment is often a long-term
therapy, establishing the long-term safety and efficacy of medications for the disease is essential.
Previously, a multinational, three-year, placebo-controlled clinical trial was extended by two
years, and that study demonstrated the sustained anti-fracture benefit of risedronate.

In the current study, 164 women continued the earlier study for two more years – a total of seven
years. All patients were treated with risedronate, even those patients who were previously
receiving placebo. Both the seven-year treatment group and the placebo-switched group
experienced increases in bone mineral density (BMD; a parameter of bone strength) in years six
and seven at the spine and hip. For the seven-year treatment group, this increase represented a
continued improvement in BMD over seven years. Also, within the placebo-switched group, the
number of vertebral fractures occurring in years six and seven dropped to a rate comparable to
that of the seven-year treatment group.

This research was funded by P&G Pharmaceuticals and Aventis Pharmaceuticals.

                                              # # #




                                                114
                                                                                                 Osteoporosis
P2-515
Adult Survivors of Pediatric Cardiac Transplantation Should Be Evaluated for Osteoporosis.
Adi Cohen*1, Linda J Addonizio2, Jacqueline M Lamour2, Elizabeth Shane1. 1Dept of Med, Div of Endocrinology;
2
  Dept of Peds, Div of Cardiol, Columbia Univ Coll of Physicians and Surgeons, New York, NY.

Adults who undergo cardiac transplantation (CTX) commonly develop bone loss and fractures. Hypothesizing that
adults who have had a CTX during childhood would also develop osteoporosis, we studied adult survivors of
pediatric CTX in a case-control, cross-sectional evaluation of bone mineral density (BMD), indices of mineral
metabolism and bone turnover markers. Nine patients (2 women), who were transplanted between ages 12-16, were
evaluated (ages 21-32) and compared to age, gender and race matched controls (C). Patients (P) had CTX at a mean
of 12±4 years prior to BMD evaluation and were treated with prednisone (mean dose at evaluation: 6±3 mg/day),
azathioprine, and cyclosporine or FK506. Mean BMD Z scores in P were -2.3±0.9 at the spine (LS), -1.4±0.6 at the
total hip (TH), -1.6±0.7 at the femoral neck (FN), and -3.2±0.7 at the distal radius (DR). BMD Z scores were
significantly higher in C (LS: -0.47±1.2, TH: 0.37±0.8, FN: 0.18±1.0, DR: 0.06±0.9; p<0.001 vs. P at all sites by
paired t test). Osteoporosis (Z-2) was present at the LS, FN and DR in 56%, 33%, and 100% of P, respectively.
Although body mass index was similar between P and C, current height compared to midparental height was
significantly lower in P (-8.2±4 vs. 0.5±8 cm; p=0.03). Bone mineral apparent density (BMAD, g/cm³), which
corrects for the effect of smaller bone size of P on BMD, remained significantly lower in P than C at the LS
(p=0.007), FN (p=0.001) and DR (p=0.003). By paired t test, parathyroid hormone (PTH) was 3-fold higher in P (P:
75±19, C: 25±7 pg/mL; p<0.001). Serum calcium, phosphorus, and 25-OH vitamin D were normal and not different
between P and C. Bone specific alkaline phosphatase (P: 35±13, C: 26±10 U/L; p=0.007), osteocalcin (P: 12±7, C:
7±2 ng/mL; p=0.06) and serum N-telopeptide (P: 30±14, C: 16±2 nM BCE; p=0.03) were higher in P than C. BUN
was higher in P (P: 30±12, C: 17±5 mg/dL; p = 0.04) and creatinine clearance tended to be lower (P: 83±42, C:
114±26 ml/min; p=0.07). Testosterone did not differ between male P and C. In summary, BMD is significantly
lower in adult survivors of pediatric CTX, particularly at the DR, a site sensitive to excessive PTH secretion. BMAD
is also lower, suggesting that bone size does not explain the difference in BMD. Low BMD is probably related to
mild renal insufficiency, hyperparathyroidism and increased bone turnover. We conclude that adult survivors of
pediatric CTX should be evaluated for osteoporosis.

CLINICAL POSTER: Osteoporosis (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                       115
                                                                                    Osteoporosis
P2-515 News Summary
Adults who had heart transplants as children should be checked for osteoporosis

Children who have a heart transplant may not acquire bone mass during the critical period of
bone growth in adolescence and, therefore, may cause osteoporosis as they age, according to a
new study being presented on Friday, June 20, at The Endocrine Society’s 85th Annual Meeting
in Philadelphia. Researchers recommend that adult survivors of pediatric cardiac transplantation
should be evaluated for osteoporosis.

Studies have shown that adults who undergo cardiac transplantation commonly develop
osteoporosis and fractures. The bone disease is thought to be related to cardiac illness as well as
to the immunosuppressive medications that patients must take after organ transplantation.

Until now, however, there have been no data on children and adolescents who receive cardiac
transplants and if they are also at increased risk for osteoporosis. In this cross-sectional study,
Dr. Adi Cohen and colleagues at Columbia University College of Physicians and Surgeons in
New York City evaluated bone mineral density and markers of bone turnover in adult survivors
of pediatric cardiac transplantation. They studied nine transplant patients who had received a
cardiac transplant before the age of 18 and were over the age of 20 at the time of the evaluation.

These transplant patients were compared to nine age-, gender- and race-matched control subjects.
The researchers found that bone density of the spine, hip and wrist was significantly lower in the
transplant patients compared to the control subjects. In fact, all of the transplant patients met
World Health Organization criteria for osteoporosis, compared to 22 percent of the control
subjects. The transplant patients were also found to have biochemical evidence of increased
bone turnover, elevated parathyroid hormone and mild renal insufficiency – possibly
contributing to the type of osteoporosis seen in the transplant recipients, say researchers. Larger
studies will be needed to assess fracture risk and determine whether there is ongoing bone loss.

This research has been funded by a grant from the Endocrine Fellows’ Foundation.

                                               # # #




                                                116
                                                                                                   Osteoporosis
P2-513
Differential Influence of Aging and Menopause on Hip Bone Loss in Healthy Middle-Aged Women.
Jane Lukacs*1, Nancy Reame1,2, Michael Kleerekoper3, Rudi Ansbacher4, Alice Rolfes-Curl5, Vasantha
Padmanabhan2,5. 1Sch of Nursing, The Univ of Michigan, Ann Arbor, MI; 2Reprod Sci Prog, Univ of Michigan,
Ann Arbor, MI; 3Endocrinology, Wayne State Univ, Detroit, MI; 4Ob and Gyn, The Univ of Michigan, Ann Arbor,
MI; 5Peds, Univ of Michigan, Ann Arbor, MI.

Although estrogen withdrawal at menopause is a major risk factor for osteoporosis, little is known about the nature
and time course of this bone loss especially in middle-aged women prior to the onset of menstrual cycle irregularity.
The objective of this study was to distinguish the effects of aging vs menopause status on bone turnover and bone
loss during the late reproductive years. We assessed bone turnover with alkaline phosphatase bone isoenzyme (ALP-
B) and bone mineral density (BMD) in regularly cycling mid-life women (CO n=13; mean age = 48 ± 0.5 yrs) and
compared them to subjects differing by age (under 30 yrs, CY n = 19; mean age 25 ± 1 yrs) or menopausal status
(PM n=13; mean age = 49 ± 1, mean yrs PM 3 ± 0.7 yrs). All cycling women were assessed on menstrual cycle day
5 ± 1. Volunteers were not using oral contraceptives or hormone replacement. Lumbar spine (L1-L4) and hip bone
mineral density (BMD) were assessed by DEXA (Lunar DPX-L). Data are expressed as mean ± S.E. Although there
were no differences in estradiol (E2), 43.7 ± 12.6 pg/ml vs 24.6 ± 2.6, p = NS), cycle length (28 ± 1 vs 31±1days, p =
NS), or bone turnover (ALP-B, 91.1 ± 7.5 vs 100.8 ± 7.2 U/L, p = NS ) between CO and CY groups, there were
differences in testosterone (T, 0.2 ± 0.03 vs 0.4 ± 0.02 ng/ml, p < 0.001), free T (0.5 ± 0.08 vs 0.9 ± 0.07 pg/ml, p <
0.001), femoral neck BMD (0.928 ± 0.038 vs 1.0723 ± 0.039 g/cm², p < 0.02) and Ward’s BMD (0.9052 ± 0.046 vs
1.0936 ± 0.051 g/cm², p < 0.02). In contrast to CO, the PM women had lower E 2 (43.7 ± 12.6 vs 9.9 ± 3.0 pg/ml, p <
0.001), similar concentrations of T (0.2 ± 0.03 vs 0.2 ± 0.03 ng/ml, p = NS), and free T (0.5 ± 0.08 vs 0.6 ± 0.1
pg/ml, p = NS); higher bone turnover (ALP-B, 91.1 ± 7.5 vs 133.8 ± 7.2 U/L, p  0.001) and corrected calcium (9.2
± 0.2 vs 9.4 ± 0.2 mmol/L, p < 0.004), and lower total hip BMD (1.0235 ± 0.041 vs 0.9035 ± 0.024 g/cm², p = 0.04).
These data suggest that in middle-aged women, aging-related bone loss has only a minor influence on bone function
compared to the impact of estrogen withdrawal which affects a number of parameters of bone turnover and bone
density.
         Pharmaceuticals of Pfizer, Inc.[br]NIH grants R01-AG15083 and 5-MORR00042

CLINICAL POSTER: Osteoporosis (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                         117
                                                                                  Osteoporosis
P2-513 News Summary
Menopausal estrogen loss affects risk of osteoporosis more than aging-related bone loss

In middle-aged women, aging-related bone loss has a minor impact on the metabolic activity of
bone compared to the impact of the estrogen loss that accompanies menopause, which affects
metabolic activity and bone density to a larger degree, according to a new study being presented
on Friday, June 20, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

Although estrogen withdrawal at menopause is a major risk factor for osteoporosis, little is
known about the nature and time course of this bone loss, especially in middle-aged women
before the onset of menstrual cycle irregularity.

By measuring the rate of metabolic activity in bone with a bone turnover marker (alkaline
phosphatase bone isoenzyme), and bone mineral density at the spine and hip (measured by duel-
energy X-ray absorptometry), Dr. Jane Lukacs and colleagues in Ann Arbor, Mich., and Detroit
sought to distinguish the effects of aging versus menopause. The researchers needed this
information to determine the timing of health interventions to maintain bone mass in healthy
middle-aged women.

They compared 13 middle-aged women (average age of 48) with regular menstrual cycles to two
control groups of women: 19 young (average age of 25) with regular menstrual cycles and 13
postmenopausal (average age of 49, three years postmenopausal). All women were of similar
body size and were not taking hormones. The women with regular menstrual cycles were
studied in the early portion of a menstrual cycle to avoid the normal hormone fluctuations
throughout the cycle. The older women with regular menstrual cycles were similar to young
cycling women with respect to estrogen status, menstrual cycle length, metabolic activity in bone
and actual calcium levels measured in the blood. However, there were two aging effects: lower
androgens (male hormones like testosterone) and less bone mass in the neck region of the hip.

In contrast to the cycling older women, postmenopausal women had markedly lower estrogen
levels, similar androgen levels, higher metabolic bone activity and higher blood calcium levels,
reflecting loss of calcium from bone. Although there were no differences in spine bone density,
there was lower bone density in all regions of the hip. Current medicines directed at slowing
metabolic bone activity – commonly prescribed for postmenopausal women to maintain bone
mass (like estrogen, bisphosphonates or selective estrogen receptor modulators) – might not be
appropriate in late reproductive women with regular menstrual cycles because the metabolic
activity of bone has not yet increased. Novel interventions prior to menstrual cycle irregularity
in middle-aged women designed to enhance bone mineral density will need further exploration.

This research was supported through a Pfizer Postdoctoral Fellowship.

                                              # # #




                                               118
                                                                                                  Osteoporosis
P2-501
Risedronate Rapidly Reduced Vertebral Fracture Risk in Elderly Postmenopausal Women.
Michael R McClung*1, Richard Eastell2, Ian Barton3, Steven Boonen4. 1Oregon Osteoporosis Ctr, Portland, OR;
2
  Hum Metab & Clin Biochem, Univ of Sheffield, Sheffield, United Kingdom; 3Procter & Gamble Pharmaceuticals,
Mason, OH; 4Ctr for Metab Bone Disease, Leuven, Belgium.

The risk of fractures vertebral and hip fractures related to osteoporosis increases with advancing age. However, data
are limited about the effectiveness of anti-remodeling agents to reduce fracture risk in elderly patients. Risedronate
(RIS) has been evaluated for the prevention and treatment of postmenopausal osteoporosis in women with ages
spanning several decades. The objective of this analysis was to determine the efficacy of risedronate in reducing
vertebral fracture risk in women with osteoporosis who were 75 years of age or older.
The analysis included more than 3000 subjects enrolled in the placebo and RIS 5 mg/day groups from the 3-year Hip
Intervention Program (HIP) and Vertebral Fracture Efficacy with Risedronate (VERT-MN and VERT-NA) studies.
Patients were included if they were 75 years of age, their baseline femoral neck bone density T-score was -2.5
and/or they had  1 prevalent vertebral fracture. All patients received 1000 mg/d calcium and, if baseline levels were
low, up to 500 IU/d vitamin D. Spinal radiographs were taken at baseline and at yearly intervals to determine
prevalent and incident vertebral fractures.
At baseline, the PBO and RIS groups were well matched for age and bone density. Three quarters of the patients had
at least one prevalent vertebral fracture, and the proportion of subjects with vertebral fractures at baseline did not
differ between the treatment groups.
Baseline Characteristics and Vertebral Fracture Incidence at 1 Year
                                      Placebo       RIS
Number                                1507          1531
Age (+/-SD)                           80 (3.9)      80 (3.9)
FN BMD T-score                        -3.0 (0.586) -2.9 (0.631)
% w/ prevalent vertebral fxs          74%           75%
Vertebral fx incidence                9.1%          3.4%
Relative risk reduction (95% CI) --                 65%* (46, 77%)
*p<0.001 vs placebo
          Compared with placebo, RIS significantly reduced the risk of new vertebral fractures by 65% at one year.
This effect was consistent across the three clinical studies. The risk new vertebral fractures was significantly
decreased by RIS over 3 years. RIS was well tolerated with a safety profile comparable to the placebo group.
          Elderly postmenopausal women with osteoporosis and previous vertebral fractures are at high risk for
having recurrent vertebral fractures. In these high-risk elderly patients, risedronate therapy produces a rapid and
sustained reduction in the risk of new vertebral fractures.

CLINICAL POSTER: Osteoporosis (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                        119
                                                                                   Osteoporosis
P2-501 News Summary
Elderly women significantly reduce risk of bone fracture with osteoporosis drug

The osteoporosis drug risedronate reduced the risk of fractures in vertebrae of the spine by 65
percent in elderly women – 75 years and older – at high risk for osteoporotic fractures, according
to a new study being presented on Friday, June 20, at The Endocrine Society’s 85th Annual
Meeting in Philadelphia. Little information was previously available about this high-risk group.

Risedronate (Actonel®) has been evaluated for the prevention and treatment of post-menopausal
osteoporosis and has been shown to be effective at preventing fractures in patients ranging in age
– spanning several decades – and disease severity. However, limited data are available on the
efficacy of osteoporosis therapies in older, higher risk patients. This information is important to
the practicing physician because rapid, effective treatment is crucial for these patients.

Therefore, Dr. Michael R. McClung and colleagues in Oregon, Ohio, United Kingdom and
Belgium examined the efficacy of risedronate in a population at high risk for vertebral fracture
based upon advanced age and on having already sustained at least one vertebral fracture. This
analysis included more than 3,000 women enrolled in placebo-controlled, phase III clinical trials
evaluating the efficacy of risedronate for the treatment of post-menopausal osteoporosis.

Research shows that one in five women with osteoporosis who sustain a vertebral fracture will
suffer another one within the next year, a process sometimes described as a “fracture cascade.”
It is critical that an osteoporosis therapy act quickly in order to help prevent further fractures
from occurring. Risedronate was effective at one year in this elderly, high-risk group of patients,
similar to the results from studies including both older and younger postmenopausal women.
This supports risedronate’s usefulness for a range of osteoporosis patients.

This research was funded by P&G Pharmaceuticals and Aventis Pharmaceuticals.

                                              # # #




                                               120
                                                                                                   Osteoporosis
P2-493
Endocrine-Orthopedics Team Approach Improves Treatment for Osteoporosis in Fragility Fracture Patients.
Elizabeth A Streeeten*1, Denise Orwig2, Tim Harrington3. 1Endocrinology, Diabetes & Nutrit, The Univ of
Maryland Sch of Med, Baltimore, MD; 2Dept of Epidemiology, The Univ of Med Sch of Med, Baltimore, MD;
3
  Rheumatol, Univ of Wisconsin, Madison, WI.

After hip fracture, the risk of subsequent fracture is high. Despite the availability of medications which reduce
fracture risk in osteoporotic patients by 50%, few (0-10%) hip fracture patients are treated with medication for
osteoporosis. Recently, the team approach to management after hip fracture has been shown to improve initiation of
osteoporosis treatment but no follow up data are available on compliance.
A Fragility Fracture Intervention Program (FFIP) was started at the University of Maryland Hospital (UMMS) and
affiliated VA one year ago to improve osteoporosis treatment and reduce subsequent fracture in patients with hip
and other inpatient fragility fractures. Patients hospitalized for fragility fracture were seen by our inpatient
Endocrine consult team, evaluated for secondary causes of osteoporosis (history, exam, lab studies) and started
during hospitalization on calcium 500 mg tid, Vitamin D 800 u qd and a bisphosphonate (weekly p.o. alendronate or
risedronate, or 30 mg IV pamidronate). A DXA was recommended after discharge. A few patients were seen shortly
after discharge. Compliance with medication was tracked by phone with the aid of “Stop Osteoporosis” software.
From 11/01-12/02, 31 patients were referred by orthopedists to the FFIP. Of these, 26 patients (80.8%) were seen (7
were seen shortly after discharge) by the endocrine consult team, 12 men (mean age 64.6±14.9) and 16 women
(mean age 69.9±13.9). Fractures were: 12 hip, 4 distal femur, 2 tib/fib, 2 tibia, 2 humerus, 2 pelvis, 2 lumbar spine.
Mean 25-OHD was 10.3±9.1 ng/ml (n=10) on inpatients (3 undetectable); 21.3±5.4 ng/ml (n=5) on those seen after
discharge. One patient was diagnosed with hemochromatosis as a result of the endocrine consult.
All patients seen were started on treatment for osteoporosis: 21/26 (81%) on calcium, Vit D and a bisphosphonate;
5/26 (19%) on calcium and Vit D. On follow up (3-11 mos), 14 patients were both available by phone and  3 mos
out from treatment initiation:11/14 (71%) patients remained on calcium, Vit D and a bisphosphonate; 3/14 (29%) on
calcium and Vit D.
Although the number of patients in this pilot study is low, we conclude that evaluating and treating fracture
inpatients for osteoporosis during hospitalization are feasible. This endocrine-orthopedic team approach may
increase the percentage of fragility fracture patients who receive osteoporosis treatment long term and reduce
subsequent fractures in this high risk population.

CLINICAL POSTER: Osteoporosis (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                         121
                                                                                   Osteoporosis
P2-493 News Summary
Team approach to evaluating fractures leads to better treatment for osteoporosis

A team approach for evaluating and treating fracture patients for osteoporosis resulted in all
fracture patients receiving some form of medical therapy for osteoporosis, according to a study
being presented on Friday, June 20, at The Endocrine Society’s 85th Annual Meeting in
Philadelphia. The researchers say that this is significant considering that previous reports show
that no more than 10 percent of fracture patients are often evaluated for the bone disease.
Furthermore, among patients who were available for follow up by phone after discharge, all
remained on some medical therapy for osteoporosis, which hasn’t previously been documented.

Osteoporosis is a condition characterized by reduced bone strength, leading to an increased risk
of fracture. Osteopenia is a milder form of osteoporosis. Osteoporosis affects approximately 30
percent of postmenopausal women and up to 6 percent of U.S. men over age 50. Even more
prevalent, osteopenia affects approximately 54 percent of postmenopausal women and up to 47
percent men over 50. One of the most feared complications of osteoporosis is hip fracture. After
hip fracture, only 25 percent of victims return to their pre-fracture mobility and independence, 25
percent die within a few months and 50 percent need nursing home care or lose independence.

Of the many risk factors for fracture, the highest risk is in those who have already had a fracture.
Essentially all patients who suffer a hip fracture from mild trauma have either osteoporosis or
osteopenia. There are several prescription and nonprescription medications that improve bone
strength significantly. Even so, the percentage of hip fracture patients who receive any treatment
for osteoporosis is very small. Without more aggressive treatment than is currently given for
osteoporosis in the United States, the number of hip fractures is expected to double or triple from
1980 to 2050. Simply initiating medical therapy for osteoporosis for a fracture could reduce the
number of these high-risk patients who fracture again by 50 percent.

Dr. Elizabeth Streeten and colleagues started a Fragility Fracture Intervention Program (FFIP) at
the University of Maryland Hospital (UMMS) in November 2001 to improve osteoporosis
treatment and reduce subsequent fractures in patients with hip and other inpatient fragility
fractures. Patients hospitalized for fragility fracture were seen by an inpatient endocrine consult
team, evaluated for underlying causes of osteoporosis and started on therapy for osteoporosis. In
one month, 26 inpatients with fractures were seen in the FFIP. There were 12 men and 14
women. Most had vitamin D deficiency. All patients seen in the FFIP were started on treatment
for osteoporosis, 81 percent on a bisphosphonate, calcium and vitamin D and 19 percent on
calcium and vitamin D alone. Compliance with medication was tracked by phone, with the aid
of a new software program, “Stop Osteoporosis,” developed by one of the authors. Of 14
patients available by phone for follow up, over a three to 11 month period, all remained on
calcium and vitamin D and 71 percent also remained on the bisphosphonate.

This study received no funding. The development of the “Stop Osteoporosis” software was
funded by Proctor and Gamble.
                                           # # #


                                                122
                                                                                                 Osteoporosis
P2-487
Small Proportion of Non-Vertebral Fracture Risk Reduction Explained by BMD Increases.
Nelson B Watts*1, D Felsenberg2, T D Johnson3, Zhengqing Li3, Richard Eastell4. 1Univ of Cincinnati Osteoporosis
Ctr, Cincinnati, OH; 2Dept of Radiology, Univ Hosp Benjamin, Berlin, Germany; 3Procter & Gamble
Pharmaceuticals, Mason, OH; 4Univ of Sheffield, Northern Gen Hosp, Sheffield, United Kingdom.

It is becoming increasingly apparent that BMD increases explain only a very small proportion of the observed
vertebral fracture risk reduction with anti-resorptive agents. In the present work, we examined the relationship
between changes in lumbar spine (LS) or femoral neck (FN) BMD and osteoporosis-related nonvertebral fracture
risk reduction in postmenopausal women.
The analysis included patients from the two risedronate VERT studies who were enrolled in the trials on the basis of
low LS BMD (T-score < -2.0) and at least one prevalent vertebral fracture or at least two prevalent vertebral
fractures. Patients were randomized to receive either risedronate 5 mg or placebo daily. Patients also received 1000
mg/day calcium supplementation and vitamin-D, if baseline levels were low.
The risk reduction by risedronate 5 mg treatment over 3 years was estimated to be 35% (95% CI=11%, 52%). The
mean BMD percent increases over placebo at endpoint were 4.6% at lumbar spine and 2.6% at femoral neck. The
analysis revealed a non-linear relationship between BMD increases and nonvertebral fracture risk reductions. The
proportion of nonvertebral fracture risk reduction explained by the BMD increases over 3 years (5mg vs. placebo)
was estimated to be 12.2% (95% CI=5.7%, 26.1%) for lumbar spine and 5.5% (95% CI=2.6%, 11.9%) for femoral
neck.
The relationship between BMD increases and nonvertebral fracture risk reduction is non-linear. BMD increases
observed during risedronate therapy only explain a small portion of the non-vertebral osteoporosis-related fracture
risk reduction. Other possible factors, which contribute to anti-fracture effects may include maintenance of bone
microarchitecture and improved bone quality.

CLINICAL POSTER: Osteoporosis (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                        123
                                                                                    Osteoporosis
P2-487 News Summary
Bone density represents small portion of osteoporosis drug’s effect on preventing fractures

Effects of an osteoporosis drug on other functions besides bone mineral density (BMD) must
account for its efficacy in preventing fractures, perhaps beneficial effects on bone remodeling,
bone microarchitecture or other aspects of bone quality, according to a new study being
presented on Friday, June 20, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

Bone mineral density (BMD) is a useful tool for the diagnosis of osteoporosis. However, based
on the study’s results, the increases in BMD after initiating treatment with osteoporosis therapies
can only explain a fraction of the observed reduction in vertebral and non-vertebral fractures.

According to the National Osteoporosis Foundation (NOF), the clinical decision to treat
osteoporosis with drug therapy should be based on strong evidence that it effectively prevents
fractures. Even though BMD is sometimes used as a surrogate marker for efficacy, it does not
have a linear relation to fracture reduction in patients who are receiving therapy. In addition, the
U.S. Food and Drug Administration (FDA) requires anti-fracture efficacy to be a primary
endpoint for osteoporosis treatment studies, and, therefore, it is the primary clinical endpoint by
which all osteoporosis treatments should be evaluated.

The data for the current analysis – conducted by Dr. Nelson Watts and colleagues from
Cincinnati and Mason, Ohio, Germany, the United Kingdom – were from the two pivotal trials,
which enrolled more than 3600 patients, supporting the approval of risedronate (Actonel®) for
the treatment of post-menopausal women for osteoporosis.

Although risedronate therapy significantly reduced the risk of non-vertebral fractures over three
years in prospective, randomized clinical trials, this analysis showed that increases in BMD
could only account for 6 percent to 12 percent of the benefit. It has become increasingly clear
that increases in BMD only explain a small fraction of the reduction in vertebral fractures with
osteoporosis therapies, and this analysis demonstrated that a similar situation exists for non-
vertebral fractures, too.

These results, say the researchers, demonstrate the importance of a physician basing his or her
therapeutic choices on the fracture reduction as demonstrated in clinical trials because reducing
fractures is the clinically relevant goal of osteoporosis therapy.

This research was funded by P&G Pharmaceuticals and Aventis Pharmaceuticals.

                                               # # #




                                                124
                                                                  Osteoporosis Diagnosis & Therapy
OR54-2
Early Changes in Biochemical Markers of Bone Formation Predict Improvements in Bone Structure during
Teriparatide Therapy.
Harald Dobnig*1, Astrid Fahrleitner1, Louis-Georges Ste-Marie2, J Chris Gallagher3, Imre Pavo4, Adrien Sipos4,
Jingyuan Wang4, Erik F Eriksen4. 1Dept of Intern Med, Univ of Graz, Graz, Austria; 2Ctr de Recherche du CHUM,
Hosp St-Luc, Montreal, QC, Canada; 3Bone Metab Unit, Creighton Univ, Omaha, NE; 4Lilly Res Labs, Eli Lilly and
Co, Indianapolis, IN.

Teriparatide [rhPTH(1-34)], a new bone formation agent for osteoporosis treatment, reversed osteoporotic changes
in bone structure (1) and decreased vertebral and nonvertebral fracture rates in a Fracture Prevention Trial (2). A
significant proportion of new bone formed during teriparatide treatment seems to be formed via modeling, i.e.
formation of new bone without previous resorption. To evaluate biochemical markers of bone metabolism as early
predictors of the effect of teriparatide therapy on new bone formation at the tissue level, we correlated the change in
markers of bone formation at 1 and 3 months with change in parameters pertaining to bone formation, assessed by 2-
dimensional (2D) histomorphometry and 3-dimensional (3D) structural µCT analysis. Thirty-one paired iliac crest
biopsies were obtained from patients treated with teriparatide for 12-24 months. At 1 month, increases (percent
change from baseline) in serum bone specific alkaline phosphatase (S-BAP) and serum C-terminal propeptide of
type I procollagen (S-PICP) were significantly correlated with the increase in 2D mean wall thickness (BAP, r=0.61,
p<0.001; PICP r=0.43, p=0.02) and 3D trabecular bone volume/total volume ratio (BAP r=0.46, p=0.009; PICP
r=0.39, p=0.03). Increases in BAP also correlated with increases in 2D trabecular bone volume (r=0.51, p=0.01), 3D
trabecular bone volume (r=0.41, p=0.02) and 3D trabecular thickness (r=0.44, p=0.01). At 3 months, S-PICP
showed an inverse correlation with marrow star volume (r=-0.44, p<0.05), an index inversely related to trabecular
connectivity. Early changes in markers of bone resorption like urinary N-terminal type I collagen cross-link peptide
(U-NTX), urinary total or free deoxypyridinoline were not correlated to changes in structural parameters. U-NTX at
3 and 12 months, however, exhibited significant correlations to tissue level (2D) mean bone resorption rate (r=0.67,
p<0.001 and r=0.73, p<0.001, respectively).
In conclusion, our data suggest that early increases in biochemical markers of bone formation (BAP and PICP)
predict improvements in bone structure during teriparatide therapy as reflected in increased mean wall thickness,
trabecular thickness, trabecular connectivity and trabecular bone volume.
         Reference: 1. Turner RT, et al. Endocrinology 1998
         2. Neer RM, et al. N Engl J Med 2001

CLINICAL ORAL: Osteoporosis Diagnosis & Therapy (2:35 PM - 3:35 PM)

Presentation Date: 6/22/2003, Time: 2:50 PM; Location: Ball B




                                                         125
                                                       Osteoporosis Diagnosis & Therapy
OR54-2 News Summary
Long-term bone strength improvements can be predicted by early markers of change
produced by new osteoporosis drug

The early changes in the biochemical markers of bone formation in patients taking a new drug
for osteroporosis, teriparatide, correlate with long-term changes in bone structure, according to a
new study being presented on Sunday, June 22, at The Endocrine Society’s 85th Annual Meeting
in Philadelphia. This correlation may provide an important early tool to predict the later bone-
building effect of the drug, say the researchers.

Osteoporosis is a major health threat, with up to one-half of women over the age of 50
experiencing one or more osteoporosis-related fractures during their lifetime. Until recently, the
only approved osteoporosis treatments were antiresorptives, which work mainly to slow or stop
bone loss by reducing the number and action of bone-removing cells called osteoclasts.

Teriparatide – recombinant human parathyroid hormone (1-34) – is a unique new bone formation
agent for the treatment of osteoporosis. Teriparatide reverses osteoporotic changes in bone
structure and decreases bone fracture rates. Teriparatide stimulates the formation of new bone
by increasing the number and action of bone-forming cells, called osteoblasts.

Histological examinations of bone biopsies are used to detect structural changes of bone.
Previously, it has been published that teriparatide results in significant improvements in bone
structure by increasing bone volume and strength after 12–24 months of treatment. Biochemical
markers of bone turnover are proteins in the blood and urine reflecting the dynamic changes in
bone metabolism through the activities of the bone-forming osteoblasts and the bone-resorbing
osteoclasts. Teriparatide results in early, rapid increase of biochemical markers of bone
formation as early as one to three months of treatment.

In the current study, Dr. Harald Dobnig of Graz, Austria, and colleagues in Graz, the United
States and Canada studied 31 biopsies of the pelvic bone from patients treated with teriparatide
for 12–24 months . They found that the early – one to three months – changes in the
biochemical markers of bone formation correlated with the long-term changes in bone structure
after 12–24 months of teriparatide treatment.

Recently, the U.S. Food and Drug Administration approved FORTEO® (teriparatide [rDNA
origin] injection) for the treatment of osteoporosis in postmenopausal women who are at high
risk for a fracture.

This study was funded by Eli Lilly and Company.

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                                                126
                                       Dietary & Hormonal Contributions to Osteoporosis
OR21-1
Idiopathic Hypercalciuria (IH) Is the Leading Cause of the Vertebral Fractures in Human Beings: A Ten-
Year Prospective Study on 2.753 Female and 153 Male Patients with IH.
Maurizio Bevilacqua*1, Laura Baruffaldi1, Velella Righini1, Luigi Foddis1, Rosanna Toscano1, Tarcisio Vago1.
1
  U.O. di Endocrinologia, Hosp Luigi Sacco-Polo Univ, Milano, Italy.

Background and Aims. Idiopathic hypercalciuria (>300 mg/die in ♂, 250 mg/die in ♀ or 4 mg/Kg/die) is the
leading cause of nephrolithiasis (~ 50%). Moreover hypercalciuric patients are also characterized by decreased
lumbar and femoral bone density. Increased (~ 4 fold) incidence of vertebral fractures has been observed in
nephrolithiasic subjects in Rochester (USA): however it is unknown if renal lithiasis or hypercalciuria “per se” are
associated to vertebral fractures. This study was designed to quantitatively investigate the relation between
hypercalciuria and fractures.
Materials and Methods. We prospectively studied the incidence of hypercalciuria in 13.259 pts consecutively,
admitted to our facilities in the decade 1991-2001.
Patients were otherwise healty men (689) and women (12.570) who attended the out patients clinic for aging-related
problems. Routine analysis included lumbar and femoral BMD (Hologic 2000 until 1998, then Delphi), lateral D 2 –
L5 x-rays, 1-84 PTH, ionized calcium, urinary NTX, 25 (OH) Vit D, alkaline phosphatase, 24 hr urinary calcium and
creatinine to exclude secondary hypercalciuria.
Results. Incidence of hypercalciuria was 22,0% in female (2.765) and 22,2% in male (153). After exclusion of
absorptive hypercalciuria (by a dietist-assisted low-calcium diet for 1 month and exclusion of patients with 2 hr
calcium/creatinine ratio < 0.36), we obtained 1.232 female (9,8% of female) and 47 male (6,8% of male) with
fasting (idiopathic) hypercalciuria.
The total number of vertebral and lumbar fractures was 687, 81 in ♂ (34 in normocalciurics: D5/12 = 25, L1/5 = 9; 47
in hypercalciurics: D5/12 = 34, L1/5 = 13) and 606 in ♀ (274 in normocalciurics: D5/12 = 173, L1/5 = 101; 332 in
hypercalciurics: D5/12 = 197, L1/5 = 135).
In female with fractures, 233 were normocalciuric (2,4% of normocalciuric female) and 262 were idiopathic
hypercalciuric (21,3% of idiopathic hypercalciuric female; p<0,001 vs normocalciuric).
In male with fractures, 16 were normocalciuric (3,0% of normocalciuric male) and 40 idiopathic hypercalciuric
(85,1% of idiopathic hypercalciuric male; p<0,001 vs normocalciuric).
Conclusion. Idiopathic hypercalciuria is a leading cause of osteoporosis.

CLINICAL ORAL: Dietary & Hormonal Contributions to Osteoporosis (1:00 PM - 2:30 PM)

Presentation Date: 6/20/2003, Time: 1:00 PM; Location: Ballroom B




                                                        127
                                  Dietary & Hormonal Contributions to Osteoporosis

OR21-1 News Summary
Calcium deficiency in the urine identifies women at risk for bone loss and fractures

Vertebral fractures are much more frequent in people affected by an excessive loss of calcium in
the urine, according to a new study being presented on Friday, June 20, at The Endocrine
Society’s 85th Annual Meeting in Philadelphia. Researchers stress that there are simple and
inexpensive measures that could identify those at risk, preventing bone loss and many of these
fractures.

Hypercalciuria – the excessive loss of calcium in the urine – affects approximately 10 percent of
the general population. It is known that people with this problem suffer from kidney stones and
the loss of bone minerals, which can make the bones brittle and susceptible to fracture.
However, the number of hypercalciuria people who have fractures has been unknown.

Therefore, research, led by Dr. Maurizio Bevilacqua, was conducted through the clinical and
laboratory evaluation of otherwise healthy people undergoing outpatient treatment for
postmenopausal and elderly-related problems at Ospedale L. Sacco in Milan. Participants also
had their bone calcium measured by densitometry.

Over a 10-year period (1991–2001), Dr. Bevilacqua and colleagues evaluated approximately
13,259 patients, identifying 153 men and 2,763 women with hypercalciuria. By measuring
urinary calcium after a short period of nutritional calcium restriction, the researchers eliminated
106 men and 1,533 women with nutrition-related–absorptive calciuria, identifying 46 men and
1,232 women with idiopathic or primary hypercalciuria – those with a slight, chronic status of
calcium loss. X-ray examination revealed that approximately 85 percent of men and 21 percent
of women with idiopathic hypercalciuria had fractures. In the general population, only 3.7
percent of women had fractures.

According to the results, idiopathic hypercalciuria increases the risk of vertebral fractures in
postmenopausal women from 3.7 to 21 percent. The small number of men examined precludes
statistical analysis. The researchers believe that a simple and inexpensive examination – 24
hours urine calcium after a short period of calcium restriction – may identify postmenopausal
women at risk for vertebral fractures. Consequently, the possibility to correct the urinary loss of
calcium, say researchers, could decrease the number of vertebral and other fractures, such as the
femur bone.

The research was partially supported by Lorenz Biotech, which sponsored two physicians
participating in the study. The majority of the study was made possible through the routine
analysis of patients admitted to the medical facilities for postmenopausal evaluation.

                                               # # #




                                                128
                                       Dietary & Hormonal Contributions to Osteoporosis
OR21-2
Prevalance of Vitamin D Deficiency among Healthy Adolescents.
Catherine M Gordon*1,2, Kerrin C DePeter1, Estherann Grace1, S Jean Emans1. 1Adolescent Med; 2Endocrinology,
Children’s Hosp, Boston, MA.

Adolescence is a critical period for bone accretion and nutritional deficits during this developmental period can
compromise the development of peak bone mass. In a clinic sample of healthy adolescent boys and girls, we sought
to determine the prevalence of hypovitaminosis D, a known risk factor for osteoporosis. We also investigated
whether a seasonal variation in serum 25-hydroxyvitamin D (25D) occurred, and if this nutritional problem was
more prevalent in girls than boys and in adolescents of a given ethnicity. 237 adolescents (85 boys and 152 girls)
who presented to an outpatient adolescent medicine practice in Boston, MA were evaluated at the time of a routine
physical examination. Serum levels of 25D, parathyroid hormone (PTH), calcium, phosphorus and magnesium were
obtained, and activity and nutritional questionnaires completed. The prevalence of hypovitaminosis D ( 15 ng/ml)
was 18% for the cohort. The problem was equally prevalent among girls (18%) and boys (18%), and more
prevalent in African American (27%) than Asian (17%), Hispanic (13%) or White (6%) teenagers. There was a
marked seasonal variation, with a prevalence of 10% noted during summer months (July - Sept.) and 47% during
winter (Jan. - March). The mean winter 25D level was significantly lower than that in summer (26.6 ± 11.2 ng/ml
vs. 17.8 ± 9.0, p =0.001), and PTH levels were higher (49.9 ± 28.2 vs. 40.0 ± 17.8 pg/ml, p=0.036). There was a
modest, but significant positive correlation between serum 25D levels and reported consumption of vitamins (r =
0.15, p = 0.22) and calcium supplements (r = 0.16, p = 0.013), and weekly exercise (r = 0.17, p = 0.011). Levels of
serum calcium, phosphorus and magnesium did not vary among seasons. There were no differences among ethnic
groups with regard to report of lactose intolerance, and consumption of milk, calcium supplements or multivitamins.
Hypovitaminosis D occurs in almost 20% of otherwise healthy teenagers in a clinic sample from Boston, a city of
Northern latitude. The problem was equally prevalent among boys and girls, and most prevalent among teenagers of
African-American and Asian descent. The positive correlation between serum 25D and specific lifestyle variables
suggests that a teenager’s vitamin D status may represent an index of adolescent wellness. As has been documented
in studies of adults and the elderly, these findings suggest that hypovitaminosis D is also a significant problem
among adolescents.

CLINICAL ORAL: Dietary & Hormonal Contributions to Osteoporosis (1:00 PM - 2:30 PM)

Presentation Date: 6/20/2003, Time: 1:15 PM; Location: Ballroom B




                                                       129
                                 Dietary & Hormonal Contributions to Osteoporosis

OR21-2 News Summary
Vitamin D deficiency is common among adolescents, affecting lifelong bone strength

Vitamin D deficiency is a common problem among adolescents, which may detrimentally affect
bone strength throughout their lives, according to a new study being presented on Friday, June
20, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

The adolescent years are critically important for the development of bone density for the rest of
life. Nutritional problems, such as vitamin D deficiency, may have a negative impact on the
bone density of growing adolescents.

To find out if this problem is widespread, researchers at Children’s Hospital in Boston, led by
Dr. Catherine M. Gordon, measured vitamin D levels in 237 healthy adolescents during routine
check-ups. Of those tested, 18 percent of the boys and girls were found to be vitamin D
deficient. The problem was equally prevalent among boys and girls, most prevalent in African
American teenagers and more common during the winter months.

The study was supported by the Charles H. Hood Foundation and the McCarthy Family
Foundation.

                                              # # #




                                               130
                                        Dietary & Hormonal Contributions to Osteoporosis
OR21-3
Serum Vitamin A and Hip Fractures: Increased Risk at Both Ends of the Concentration Curve.
Alexander R Opotowsky*1, John P Bilezikian1,2. 1Dept of Med, Coll of Physicians and Surgeons Columbia Univ,
New York, NY; 2Dept of Pharmacol, Coll of Physicians and Surgeons Columbia Univ, New York, NY.

Both vitamin A deficiency and hypervitaminosis A can adversely affect bone metabolism. Recent studies have
shown that high vitamin A intake can increase the risk of hip fracture, but these studies focused on populations
whose vitamin A intake was generally high. Intake in these studies did not reflect the broad distribution seen in the
general population. It is unclear, therefore, whether the results apply to a more general population. It seemed
possible that both high and low serum vitamin A might predispose to hip fracture, the hypothesis that we tested in
this study. The NHANES I Epidemiologic Follow-up Study, conducted between 1971 and 1974, included 2,799
women 50 to 74 years old, of whom 172 had incident hip fractures during the 22-year follow-up (14.7±5.8). The
relative risk (RR) of hip fracture for each quintile of baseline serum vitamin A, compared with the middle quintile,
was determined by Cox regression analysis. There were 39 (7.0%), 32 (5.5%), 21 (3.9%), 37 (6.3%), and 43 (7.9%)
incident hip fractures in the lowest to highest quintiles. Dietary vitamin A intake was not analyzed, since
information from NHANES was insufficient to accurately estimate vitamin A intake. When the data were controlled
for age, those in the lowest and highest quintiles had twice the risk of sustaining a hip fracture (RR=2.00, 95%CI:
1.18-3.42 and 2.02, 95%CI: 1.20-3.40 respectively, p0.01 for each) than those in the middle quintile of serum
vitamin A levels. Those in the second and fourth quintiles had a smaller and insignificant increase in hip fracture
risk (RR=1.50, 95%CI: 0.86-2.60, p=0.15 and 1.53, 95%CI: 0.89-2.61, p=0.12) The results remained significant in
the highest and lowest quintile groups after controlling for weight, race, hormone replacement use (ever vs. never),
smoking (current, former, never), level of recreational activity, dietary calcium intake, and history of previous
fractures. Of note, there was an interaction between smoking and serum vitamin A quintile. Both high and low
serum vitamin A increased hip fracture risk more significantly among current smokers than among former or never
smokers. Serum vitamin A levels are related to dietary intake of vitamin A, especially at the extremes. This is the
first study to demonstrate that both ends of the serum vitamin A concentration curve are associated with increased
hip fracture risk, and the results caution against both excess and deficient dietary intake.

CLINICAL ORAL: Dietary & Hormonal Contributions to Osteoporosis (1:00 PM - 2:30 PM)

Presentation Date: 6/20/2003, Time: 1:30 PM; Location: Ballroom B




                                                        131
                                 Dietary & Hormonal Contributions to Osteoporosis
OR21-3 News Summary
Too much and too little vitamin A is bad for bone strength, with increased fracture risk

Researchers have found, for the first time, that U.S. women with too low and too high levels of
Vitamin A are at risk for increased hip fracture, according to a study being presented on Friday,
June 20, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

Vitamin A plays an essential role in an array of biological functions. From the developing fetus
to the elderly adult, both vitamin A deficiency and excess can wreak havoc on human health.
Even moderately high or low intake of this vitamin can cause significant health effects, such as
birth defects. In adults, a vitamin A deficiency can impair the body’s ability to fight off
infection, harm reproductive function and, most important, lead to poor vision and blindness. In
addition, vitamin A status at the extremes can affect bone health. Early animal experiments
suggested that both too little and too much dietary vitamin A adversely affect bone strength;
however, because the animals were exposed to extremes of the vitamin A scale, the applicability
to the general human population at normal vitamin A intake was unclear.

Recent studies have found that individuals with high vitamin A intake have lower bone density
and are at a higher risk of fractures. Of special concern was that the levels of vitamin A intake
required to increase fracture risk was not much higher than the recommended daily intake.
These recent studies, however, focused on groups with generally high vitamin A intake, such as
in Sweden, known for eating vitamin A–rich foods like cod liver oil and dairy products.

Dr. Alexander R. Opotowsky and John P. Bilezikian of the College of Physicians and Surgeons
at Columbia University in New York City aimed to address the questions of whether older
women with vitamin A levels at the high or low ends of normal in the general population might
be at increased risk of hip fracture. They looked at a U.S. government study, started in the
1970s. Each of 2,799 women had vitamin A measured at when the study began. Over the 22
year follow-up period, 179 of these women had at least one hip fracture. They found that those
with the highest vitamin A levels were about twice as likely as those with average vitamin A
levels to sustain a hip fracture, agreeing with recent studies. However, they also found that
women in the lowest vitamin A group had almost twice the risk of having a hip fracture as
women in the middle group. This validated the researchers’ concerns that recent studies might
have missed such an effect at low vitamin A intake.

These results call for a reassessment of recent appeals for decreased vitamin A supplementation
and fortification among the general population, say the researchers. For some individuals, such
supplementation may indeed increase fracture risk. However, others with low blood levels may
benefit from extra vitamin A. This analysis suggests that, in terms of bone, small amounts of
extra vitamin A may not make much difference in overall hip fracture rates at current U.S.
vitamin A blood levels. However, elimination of vitamin A supplementation and fortification
could conceivably increase the number of vitamin A–deficient individuals. Such individuals
would also be at increased risk for vitamin A deficiency’s other related health effects.
                                               # # #


                                               132
                                                        Parathyroid Hormone, Calcium & Bone
P1-351
Estrogen Treatment Mitigates but Does Not Prevent Bone Loss during Lactation.
Joshua VanHouten*1, Pamela Dann1, John J Wysolmerski1. 1Intern Med/Endocrinology and Metab, Yale Univ Sch
of Med, New Haven, CT.

The most rapid period of bone loss in adults occurs during lactation. However, the regulation of this phenomenon is
incompletely understood. We studied separate cohorts of age-matched CD-1 mice on days 2, 5, 12, and 18 post-
partum. Controls were age-matched and included virgins and mice at the end of pregnancy. BMD, as assessed by
DEXA, progressively declined over the first half of lactation, and was 33% lower at the spine, 22% lower at the
femur and 21% lower for total body at day 12. Urine N-telopeptide (NTx) levels were elevated during lactation but
plasma osteocalcin levels were suppressed, suggesting that bone turnover was uncoupled. Plasma calcium levels
were slightly higher during pregnancy and lactation, but PTH levels remained unchanged. We next examined the
effects of dietary calcium restriction and estrogen and pamidronate treatment on bone metabolism during lactation.
Mice were placed on either normal (0.6% Ca) chow or a low calcium diet (0.01% Ca) upon delivery, and BMD was
measured at day 4 and then day 12 of lactation. Calcium restriction approximately doubled the rate of bone loss at
the spine, femur and total body. This was associated with a significant increase in urine NTx levels and maternal
hypocalcemia and secondary hyperparathyroidism. Estrogen levels were low in lactating controls, confirming an
estrogen-deficient state. Estradiol treatment was effective at raising circulating levels of estrogen, increasing
uterine weights and normalizing urine NTx levels. It mitigated but did not eliminate bone loss, reducing the change
in BMD by approximately 50% at all sites. Pamidronate treatment also normalized urine NTx levels and reduced
the rate of bone loss by 40-50%. We conclude that mice lose bone rapidly during lactation. Bone loss is worsened
by dietary calcium restriction and is lessened, but not prevented, by estrogen or bisphosphonate treatment. Our data
suggest that lactation is a state of physiologic uncoupling of bone turnover characterized by increases in bone
resorption and decreases in bone formation. Although estrogen deficiency triggers increased rates of bone
resorption during lactation, suppression of bone formation is likely to play an equally important role in mediating
the rapid mobilization of skeletal calcium necessary for milk production.

BASIC POSTER: Parathyroid Hormone, Calcium & Bone (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                        133
                                                 Parathyroid Hormone, Calcium & Bone
P1-351 News Summary
Researchers uncover the mechanism of temporary bone loss in breastfeeding women; may
lead to treatments for osteoporosis

The mechanism behind temporary bone loss during lactaction is based on estrogen levels and the
body’s suppression of bone formation, according to a study being presented on Thursday, June
19, at The Endocrine Society’s 85th Annual Meeting in Philadelphia. The goal of the research,
say the investigators, is to lead to new ideas for osteoporosis treatments.

Breastfeeding women undergo a rapid and substantial loss of bone density, which rebounds
rapidly after breastfeeding is discontinued. This bone loss is probably a mechanism for releasing
the calcium, stored in bones, that is needed to make milk. After lactation, bone mass is
subsequently recovered with no apparent ill effects.

Based on this knowledge, Joshua N. VanHouten and colleagues at Yale University School of
Medicine in New Haven, Conn., systematically examined bone loss during lactation using a
mouse model. They measured bone density by dual energy X-ray absorptiometry in groups of
non-mated, pregnant, lactating and weaned mice. They found that by mid-lactation, bone
mineral density declined by approximately 20 percent to 30 percent.

There is normally a balance between bone resorption – the process of breaking down existing
bone to free stored calcium – and the formation of new bone. When bone resorption increases,
so does bone formation, so there is usually not a rapid net gain or net loss of bone density. A key
finding of this study, say researchers, is that during lactation these processes become unbalanced.
By measuring biochemical markers of bone resorption and bone formation in lactating mice, they
found that bone formation is decreased while bone resorption is elevated. This phenomenon is
known as “uncoupling” of resorption and formation.

They next tried to find the hormones involved in regulating bone resorption and formation during
lactation. Because estrogen levels are usually low in lactating mammals, the prevailing
hypothesis has been that the lack of estrogen may be responsible for bone loss during lactation,
like during menopause. Artificially elevating estrogen levels in lactating mice with daily
injections slowed, but did not completely prevent bone loss.

Although estrogen replacement reversed the elevation in bone resorption in lactating mice, it
failed to prevent the decrease in bone formation. Therefore, the rapid removal of calcium from
the skeleton for milk production during lactation is due, in part, to increased bone resorption that
is permitted by estrogen deficiency. Suppression of bone formation, which was not stopped by
estrogen, is likely to be equally important in bone loss during lactation.

This research is supported by grants from the National Institutes of Health.

                                               # # #



                                                134
Pediatric Endocrinology
                                                             Pediatric Endocrinology: Growth & Puberty
OR8-4
Intrauterine Growth Retardation Versus Prematurity as Determinants of Reduced Insulin Sensitivity in
Childhood.
Rodrigo A Bazaes*1, Angelica Alegria2, Alejandra Avila1, German Iniguez1, Veronica Mericq1. 1Inst of Maternal
and Child Res, Univ of Chile, Santiago, Chile; 2Neonatal Unit, Dr. Sotero del Rio Hosp, Santiago, Chile.

Low birth weight (LBW) is associated with decreased insulin sensitivity in postnatal life. Available data come from
retrospective studies, assessing birth weight regardless of gestational age. Our goal was to study whether some of the
risks observed in LBW subjects may be related to prematurity rather to an adverse in utero environment.
We studied insulin secretion and sensitivity in 60 premature, very LBW children aged 5-7 years (birth weight <
1500 g, gestational age 25 to 34 weeks). Twenty (33%) were small for gestational age (SGA), and 40 (67%) were
appropriate for gestational age (AGA). Early postnatal morbidity was similar in both groups. A short IVGTT was
performed after an overnight fast by infusion of 0.5 g/kg glucose over 3 minutes. Blood glucose and serum insulin
were measured at times –5, 0, 1, 3, 5 and 10 min. Serum levels of IGFBP-1, SHBG and lipid profile were
determined in the basal samples. Fasting insulin sensitivity was estimated using HOMA, and first phase insulin
secretion was calculated as the insulin area under the curve (AUC). Data are shown as median (interquartile range),
and were analyzed using non parametric statistics. ANCOVA was also used in order to allow for the confounding
effects of current adiposity (assessed as BMI) and gestational age.
                              SGA (n=20)          AGA (n=40)            p (Mann Whitney’s U)
Birthweight (g)               1130 (940/1420)     1240 (1030/1430) 0.40
Gestational age (weeks)       31 (29/34)          28 (27/30)            <0.01
Current BMI (SDS)             0.91 (-0.94/2.10) 0.31 (-0.47/2.06) 0.95
Fasting insulin (uIU/ml) 5.19 (2.81/8.72)         3.79 (3.26/5.31)      <0.05
HOMA                          1.22 (0.52/1.76)    0.81 (0.65/1.12)      0.05
Insulin AUC (uIU·min/ml) 458.2 (301.5/763.9) 421.5 (260.2/651.3) 0.33
IGFBP-1 (ng/ml)               81.8 (65.4/99.3)    84.4 (68.7/98.2)      0.59
SHBG (nmol/l)                 80.8 (63.3/111.7) 81.4 (67.5/98.5)        0.92
Fasting insulin sensitivity was lower in the SGA group compared to AGA, whereas post-load insulin secretion,
IGFBP-1 , SHBG and lipid profiles were similar in both groups. Differences persisted after adjustment for current
BMI and gestational age.
These results suggest that reduced insulin sensitivity in children born small for gestational age is not attributable to
the consequences of prematurity and early postnatal morbidity, and may thus result from an adverse prenatal
environment.
          Supported by FONDECYT 2010049 and Fundacion Andes, Chile

CLINICAL ORAL: Pediatric Endocrinology: Growth & Puberty (1:00 PM - 2:30 PM)

Presentation Date: 6/19/2003, Time: 1:45 PM; Location: 204 A




                                                         135
                                           Pediatric Endocrinology: Growth & Puberty
OR8-4 News Summary
Smaller-than-expected babies, regardless of prematurity, have higher risk of adult diabetes

Being small at birth is a risk factor for the developing a resistance to insulin – a precursor to
diabetes – independent of prematurity, according to a new study being presented on Thursday,
June 19, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

Diabetes, alongside a number of related diseases, is now taking place as one of the main threats
to public health in the 21st century. During the last two decades, the remarkable increase in the
number of people diagnosed with diabetes seems to be closely linked to the widespread adoption
of a “westernized” lifestyle, and the ever-growing prevalence of obesity. Moreover, resistance to
the actions of insulin, a condition that usually precedes the onset of diabetes, has been found in
an increasing number of young people, including children.

Moreover, it has been recently recognized that other conditions may be related to the
development of diabetes, some arising – as found by Dr. David J.P. Barker – very early in life,
even before birth. Dr. Barker found that, among people born during the first third of the 20th
century, those who were smaller at birth had a higher risk of diabetes and heart disease in
adulthood. He suggested that these smaller babies were exposed to reduced availability of
nutrients during fetal life, which would result in metabolic changes leading to the development
of disease. However, a significant number of these small babies may have been actually
premature, and some researchers have raised concerns as to whether the increased risks are
related to the consequences of prematurity rather than to failure to grow in utero.

To address this question, Dr. Rodrigo A. Bazaes and colleagues at the University of Chile and
the del Rio Hospital, both in Santiago, Chile, studied 60 children, 5 to 7 years old, who were all
born prematurely. Although all of them weighed less than 1500 grams, approximately 3 pounds,
at birth, 20 were “small for gestational age” (SGA), meaning that they were smaller than
expected for the duration of pregnancy. The remaining 40 were still small, but “appropriate for
gestational age” (AGA). The researchers found early indicators of resistance to the actions of
insulin in the SGA children, as compared to their AGA counterparts. Interestingly, the degree of
resistance was not related to the duration of pregnancy, but rather to the adequacy of birth weight
to the length of pregnancy.

This research, say the investigators, will help in identifying people with a higher risk of diabetes.
More important, it has been recently shown that it is often possible at least to delay the onset of
this disease by introducing fairly simple changes in lifestyle. Focusing preventive actions in
people at higher risk will be essential in reducing the burden that diabetes and its complications
represent for society.

This research was funded by a doctoral grant from Fondecyt, Chile.

                                               # # #



                                                136
                                                                             Pediatric Endocrinology II
P2-381
Treatment of PCOS in Adolescents with Metformin.
Tracey Bridger*1, Celia Rodd2. 1Ped Endocrinology, Mem Univ, St John’s, NF, Canada; 2Ped Endocrinology,
McGill Univ, Montreal, QC, Canada.

Polycystic ovarian syndrome (PCOS) is a common disorder in females, often causally associated with
hyperinsulinemia. Metformin, an insulin sensitizer, has been used to treat this disorder in adult women with
significant improvement in fertility, hormonal imbalances and metabolic disturbances. PCOS is now increasingly
found in adolescent females
The aim of our study was to assess the effects of 3 months of metformin therapy on the androgen and insulin levels,
lipid profile, and clinical status in hyperinsulinemic non-diabetic adolescent females with PCOS compared to the
effects of a placebo. Subjects (n = 22) were randomly assigned to take either metformin 750 mg bid or placebo in a
double-blind fashion for 12 weeks. All subjects also received the same healthy lifestyle advice prior to starting the
medication. Pre- and post-treatment OGTT, fasting lipid profiles, and clinical measurements (BMI, hirsutism scores,
BP) were obtained.
At the start, the young women (age 14 to 18 years) were moderately obese (BMI 32.1 + 5.3), had elevated serum
levels of testosterone (6.3 ± 1.7 nmol/L N: <3, 0.18 ± .05 ng/dl), were hirsute and oligomenorrheic. The average
level of insulin during the OGTT was elevated (910.85 + 510 pmol/L). With metformin treatment, the insulin area-
under-the curve (AUC)significantly decreased by 30% (107689 vs 76050, p= 0.05) and the testosterone level
decreased by 30% (p= 0.03). Those given placebo had no significant change in insulin or testosterone levels. In
addition, the metformin seemed to improve the regularity of menses. No changes were noted in BMI, hirsutism
score, BP, or lipid profiles in either group. Side effects were minimal in the metformin group.
In conclusion, metformin treatment reduced hyperinsulinemia and hyperandrogenemia in adolescent females with
the hyperinsulinemic form of PCOS, with minimal side effects.

CLINICAL POSTER: Pediatric Endocrinology II (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                        137
                                                                   Pediatric Endocrinology II
P2-381 News Summary
Diabetes medicine shown to help adolescents with hormone problem and prevent diabetes

The diabetic medicine metformin can reduce high insulin and male hormone levels and improve
menstrual regularity in adolescents with polycystic ovary syndrome (PCOS), a common disorder
in women, according to a new study of being presented on Friday, June 20, at The Endocrine
Society’s 85th Annual Meeting in Philadelphia. In addition, this treatment can potentially
prevent or delay of the development of adult-onset, or type-2, diabetes.

PCOS is often characterized by excess facial hair (hirsutism), acne, menstrual irregularities,
obesity and infertility. It is diagnosed in at least 5 percent to 10 percent of women between
adolescence and menopause. Once thought to be purely a cosmetic and fertility problem, it is
now known that women with PCOS are at risk of developing diabetes, high blood pressure,
elevated cholesterol and heart disease. It is also known that PCOS can start at a young age,
many being younger than 15 years old.

No one knows exactly how PCOS starts, but high insulin levels seem to have a role. The
pancreas will make more insulin when the body’s cells do not respond to a normal amount of
insulin. The elevated insulin levels are a sign that the patient has insulin resistance or is one step
away from developing diabetes. The excess insulin is felt to alter normal hormone production in
the ovaries, leading to excess androgen – male hormones – production. In the past, the birth
control pill was generally prescribed to regulate periods and control hair growth and acne.

With the evidence of insulin resistance, treatments typical of diabetes have been investigated as
possible treatments in women with PCOS. Metformin is a medication that helps to increase the
body’s sensitivity to insulin, thereby lowering the insulin levels in the blood. It has been shown
to be an effective treatment in studies of adult women with PCOS. There are currently few
studies – and no clinical trials – showing that similar treatments work in young girls.

Therefore, Canadian researchers, Drs. Tracey Bridger and Celia Rodd, studied 22 adolescent
girls (14–18 years old), who were randomly assigned to take either metformin or placebo in a
double-blind fashion for 12 weeks. Pre- and post-treatment insulin and glucose levels, lipid
profiles and clinical measurements (body mass index, hirsutism, blood pressure) were obtained.

At the start, the young women were moderately obese, had elevated levels of testosterone and
insulin, were hirsute and oligomenorrheic (fewer than six periods per year). With metformin
treatment, the average insulin level decreased by 35 percent and the average testosterone level
decreased by 30 percent. Those given placebo had no significant change in insulin or
testosterone levels. In addition, the metformin seemed to improve the regularity of menses.
Both obese and non-obese subjects benefited from metformin therapy. No changes in other
measurements were noted in either group, and side effects were minimal.

The research was partially funded by the Montreal Children’s Hospital Research Institute.
                                            # # #


                                                 138
                                                                            Pediatric Endocrinology II
P2-385
Insulin Sensitization Early Post-Menarche Prevents Progression from Precocious Pubarche to Polycystic
Ovary Syndrome in Non-Obese Girls Born Small for Gestational Age.
Lourdes Ibanez*1, Ken Ong2, Rakesh Amin2, Maria Victoria Marcos3, David B Dunger2, Francis E de Zegher4.
1
  Endocrinology Unit, Hosp Sant Joan de Deu; Univ of Barcelona, Esplugues, Barcelona, Spain; 2Peds, Univ of
Cambridge, Cambridge, United Kingdom; 3Endocrinology, Hosp de Terrassa, Terrassa, Barcelona, Spain; 4Peds,
Univ of Leuven, Leuven, Belgium.

Girls born small for gestational age (SGA) and with history of precocious pubarche in childhood (PP), are at high
risk of polycystic ovary syndrome (PCOS) in adolescence, even if non-obese. Hyperinsulinemic insulin resistance
during the critical early post-menarcheal years may be a key factor in the development of PCOS. In formerly SGA-
PP girls, we examined whether early initiation of insulin sensitization therapy during early post-menarche could
prevent progression of PCOS features.
Twenty four girls, who were small at birth (mean birthweight: 2.4 Kg), presented with PP (at mean age: 6.7 y), and
were currently (mean age: 12.4 y) post-menarcheal, with hyperinsulinemic hyperandrogenemia but non-obese, were
randomized to receive metformin [850 mg/d] or no treatment for 12 mo. Six-monthly fasting blood samples were
collected and body composition assessed by dual-energy X-ray absorptiometry (DXA). Overnight GH and
gonadotropin secretion was analysed in 10 girls (6 treated; 4 untreated) at 0 and 6 mo.
At baseline, compared to healthy controls, SGA-PP girls with hyperinsulinemic hyperandrogenism had
dyslipidemia, excess truncal fat, reduced lean body mass, and also increased IGF-1 levels and hypersecretion of GH,
but not gonadotrophins. In untreated girls, insulin sensitivity, serum androgens, LDL- and HDL-cholesterol, total
and truncal body fat mass, and lean mass significantly diverged further from normal over 12 mo. In metformin-
treated girls, all these abnormalities significantly reversed within 6 mo, and body composition continued to improve
between 6-12 mo.
In conclusion, we report the first evidence that early metformin therapy prevents progression from PP to PCOS in
formerly SGA girls. This confirms the key role of hyperinsulinemic insulin resistance in the ontogeny of PCOS.
Furthermore, normalisation of body composition, lipid profiles and GH secretion could reduce long-term
cardiovascular risk.

CLINICAL POSTER: Pediatric Endocrinology II (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                       139
                                                                 Pediatric Endocrinology II
P2-385 News Summary
Diabetic drug shown, for the first time, to prevent endocrine condition in young women

Adolescents who were born small and exhibit certain characteristics before puberty are likely to
develop a common endocrine problem – polycystic ovarian syndrome (PCOS) – but it can be
prevented by identifying these girls at risk and administering the diabetic drug metformin,
according to first-time findings from a study being presented on Friday, June 20, at The
Endocrine Society’s 85th Annual Meeting in Philadelphia. By improving the action of insulin,
the drug not only prevented the progression to PCOS but it restored a more normal, “feminine”
body shape, characterized by a low waist-to-hip ratio.

PCOS is the most frequent endocrine condition affecting the health of young women. Symptoms
include fertility problems or the development of unwanted body hair, but a key factor in its
development is impaired action of the hormone insulin, which is a risk factor for diabetes and
heart disease. Researchers have discovered that the risk for PCOS can be identified in young
women, even before they start their periods, when low birth weight is followed by rapid
childhood weight gain, the development of abdominal fat and the occurrence of pubic hair before
puberty.

In such adolescents, researchers from Spain, United Kingdom and Belgium, led by Dr. Lourdes
Ibañez, studied 24 adolescent girls who had a history of low birth weight and developed pubic
hair around the age of 6 to 7 years old. All had experienced their first menstrual period between
6–12 months before the start of the study. Although not obese, they did have an excess of
abdominal fat and high serum insulin levels suggestive of reduced insulin action.

During the course of 12 months treatment with metformin, insulin levels came down and levels
of lipids in the blood reverted to normal. Toward the end of treatment, definite improvements in
the distribution of body fat were observed with reductions in abdominal fat. In the 12 girls who
received active treatment, there was no progression to PCOS, whereas control subjects who were
not given treatment developed symptoms and biochemical features of the condition.

The pathway to PCOS has been recognized to start before puberty and to definitely worse after
completion of puberty, say the researchers, making it clear that prevention is the best alternative
for this condition, reaping long-term benefits. They add that early intervention may be indicated
in young women at high risk of developing this condition. This is the first study pointing to the
possibility of PCOS prevention, an important public health issue.

This study was supported by the Fondo de Investigaciones de la Seguridad Social and the
Ministerio de Sanidad y Consumo of Spain.

                                              # # #




                                                140
                                                                             Pediatric Endocrinology II
P2-390
Spinal Changes Are Frequent in Pediatric Patients with Glucocorticoid-Induced Osteoporosis.
Flavia Rabie1, Outi Makitie*1, Andrea Doria2, William G Cole3, Ronald Laxer4, Alan Daneman2, Etienne B
Sochett1. 1Endocrinology; 2Diag Imaging; 3Orthopaedics; 4Rheumatol, The Hosp for Sick Children, Toronto, ON,
Canada.

BACKGROUND: Osteoporosis is one of the most serious complications of long-term systemic glucocorticoid (GC)
treatment. Risk of fracture increases with higher doses and longer duration of therapy. Spinal compression fractures
are common in GC treated adults but little is known about their prevalence in pediatric patients treated with oral
GCs.
AIM: To assess frequency of spinal changes in children with osteoporosis secondary to long-term oral GC treatment.
PATIENTS: Seventeen patients with a history of long-term oral GC treatment for chronic illness were included in
the study. All the patients had been referred to the Bone Health Clinic, The Hospital for Sick Children, for suspected
osteoporosis.
METHODS: Medical records were reviewed for details of the underlying condition and treatment. The average daily
dose of GCs over the total treatment period was estimated by dividing the cumulative dose of predinisone by the
duration of treatment time (data not available for 3 patients). Results of bone mineral density (BMD) assessments
were reviewed. Thoraco-lumbar spine radiographs were reviewed for presence of vertebral body changes by the
Kleerekoper method. These were classified as normal, mild, moderate or severe.
RESULTS: The mean age of the 17 patients (8 females) was 13.8 yrs (6.9-17.5 yrs). The mean duration of GC
treatment was 3.7 yrs (range 0.5-12.5 yrs) and the average Prednisone dose 9.3 mg/d (range 3.7 - 21.9 mg/d); the
cumulative Prednisone doses ranged from 1.4 to 55.8 g (mean 14.4 g). The mean BMD z-score for lumbar spine
(L2-L4) was -3.0 SDS (range -1.1- -8.0 SDS). Review of the radiographs showed severe spinal changes in two and
moderate changes in four patients. Three patients had mild changes and eight patients, normal spinal X-rays. The
means for cumulative Prednisone dose and BMD z-score were 28.5 g and -4.0 SDS, respectively, for patients with
moderate to severe spinal changes, and 8.7 g (P<0.05) and -2.5 SDS (P=0.077), respectively, for patients with mild
or no changes in radiographs.
CONCLUSIONS: This study suggests that compression fractures (i) are common in pediatric patients treated with
oral GCs, and (ii) are more commonly seen in patients with high cumulative GC dose and low BMD. These findings
question whether pharmacological treatment of osteoporosis should be considered to prevent spinal compression
fractures in patients with long-term oral GC treatment.
         Supported by ESPE Research Fellowship, sponsored by Novo Nordisk A/S (to O.M.)

CLINICAL POSTER: Pediatric Endocrinology II (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                        141
                                                                 Pediatric Endocrinology II
P2-390 News Summary
Long-term steroid treatment in children affects bone strength

Long-term steroid treatment has more serious effects on the bone health of chronically ill
children than previously thought, according to a study being presented on Friday, June 20, at The
Endocrine Society’s 85th Annual Meeting in Philadelphia. The researchers suggest that the
current prevention strategies are ineffective and need to be reviewed.

There is growing awareness among pediatricians that children with chronic illnesses are at risk
for osteoporosis. While the reasons are complex and not fully understood, the illness itself as
well as medications used to treat the illness seem to play major roles. Steroids are often used in
high doses for prolonged periods in children with great improvement to the underlying illness,
such as asthma. However, steroids have been found to cause osteoporosis in adult patients; by
comparison, little is known about how much they affect the bones of growing children.

In the current study, Dr. Etienne Sochett and colleagues at The Hospital for Sick Children in
Toronto examined bone health using bone mineral density (BMD) and spinal radiographs in
children who have been treated with steroids for various chronic illnesses. They studied 17
patients – from 6–17 years old – seen at the hospital’s Pediatric Bone Health Clinic. They had
been treated with steroids for six months to 12.5 years. BMD was low in most of these patients.
Spinal radiographs showed spinal compression fractures in six patients (35 percent). These six
had, on average, taken more steroids over the time of their illness and had lower BMD than
patients without compression fractures.

This study was supported by a Research Fellowship from the European Society for Pediatric
Endocrinology and Novo Nordisk.

                                              # # #




                                               142
                                                                            Pediatric Endocrinology II
P2-394
Effects of Smoking in Pregnancy on Growth in the First Year of Life.
Peter C Hindmarsh*1, Jane Pringle1, Michael P Geary1, Charles H Rodeck2, John C Kingdom3. 1London Ctr for Ped
Endocrinology, Univ Coll London, London, United Kingdom; 2Dept of Ob, Univ Coll London, London, United
Kingdom; 3Samuel Lunefeld Inst, Mt Sinai Hosp, Toronto, Canada.

Cigarette smoking is associated with a reduction in birth weight, length and head circumference. Little is known of
the growth of infants born to mothers who smoked during pregnancy. Rapid growth during the first year of life
particularly weight gain has been associated with an increased risk of cardiovascular disease in adulthood. We have
previously demonstrated that birth size and cord IGF-1 concentrations are lower in babies whose mothers smoked in
pregnancy and now describe a more detailed account of body composition at birth and at 1 year of age in the babies
of 1650 Caucasian mothers with a known smoking history. All pregnancies were singleton and analysis was
confined to pregnancies delivering at term. Pregnancies complicated by factors known to be associated with chronic
hypoxia, antepartum haemorrhage, pregnancy induced hypertension and preeclampsia were excluded along with
pregnancies where uterine blood flow was compromised yielding a sample population of 1092. At birth and at 6 and
12 months of age measures of weight, length, head circumference, skinfold thickness and mid arm circumference
were undertaken.
Status              Birth Weight SDS Change in Weight SDS at 6 months Change in Weightt SDS at 12 months
Non-Smoker          0.18               -0.11                               -0.09
Stopped Smoking 0.04                   0.03                                0.17
<10 per day         -0.28              0.55                                0.40
10-20 per day       -0.20              0.19                                0.22
>20 per day         -0.24              -0.04                               0.57
P value             <0.001             <0.001                              0.002
There was no significant change in length or head circumference over this period of time. In multiple linear
regression analysis weight gain was related to the numbers of cigarettes smoked (P<0.001) and maternal pre-
pregnancy weight (P=0.02). The higher the number smoked the longer the time taken for weight increase to be
manifest.
These results demonstrate an effect of smoking on post-natal growth with a catch-up in weight over the first year of
life with no significant change in length and head circumference. The greater the number of cigarettes smoked the
longer the time to attain catch-up. The more marked increase in weight during the first year of life in infants of
mothers who smoked may place them at increased risk of developing cardiovascular risk factors independent of their
potential smoking habits.

CLINICAL POSTER: Pediatric Endocrinology II (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                       143
                                                                 Pediatric Endocrinology II

P2-394 News Summary
Smoking During Pregnancy Increases Infants Risk of Heart Disease Later in Life

Smoking during pregnancy leads not only to a reduction in birth weight, length and head
circumference but also alters metabolism leading to weight gain and increasing the risk of heart
disease as an adult, according to a new study being presented on Friday, June 20, at The
Endocrine Society’s 85th Annual Meeting in Philadelphia.

Little is known of the growth of infants born to mothers who smoked during pregnancy. Rapid
growth during the first year of life, particularly weight gain, however, has been associated with
an increased risk of heart disease in adulthood. As a result, babies from families that have
smokers may be at risk of heart disease as a result of the smoking background itself as well as
from the effects of smoking on growth.

To further investigate, British and Canadian researchers, led by Dr. Peter C. Hindmarsh of
University College London, studied the growth of babies – whose mothers smoked in pregnancy
– during the first year of life. The study group of 1,092 mothers with a known smoking history
was selected from a larger group of 1,650 Caucasian mothers. Pregnancies complicated by other
medical conditions known to influence growth were excluded. All pregnancies were single
births, and the analysis was confined to pregnancies delivering at term. Babies born to mothers
who smoked during pregnancy were lighter, shorter and had reduced head growth at birth than
those born to mothers who did not smoke or who stopped in early pregnancy. During the first
year of life, the babies of mothers who smoked in pregnancy showed a marked increase in
weight with no change in length and head growth. In other words, they became fatter. The
effect was directly related to the numbers of cigarettes smoked.

These results demonstrate an effect of smoking on post-natal growth with a catch-up in weight
over the first year of life with no significant change in length and head circumference. The
marked increase in weight during the first year of life in infants of mothers who smoked may
place them at increased risk of developing cardiovascular risk factors independent of their
potential smoking habits. This suggests that the effects of smoking on reducing organ growth,
length and brain are permanent and that changes also take place in the metabolism of these
individuals leading to weight gain.

This research was supported by the British Heart Foundation.

                                              # # #




                                               144
                                          Pediatric Endocrinology: Fat & Insulin Sensitivity
OR42-5
Prepubertal Premature Children Are Insulin Resistant and Have an Abnormal GH/ IGF-1 Profile.
Paul L Hofman*1, Fiona M Regan1, Wendy E Jackson1, Elizabeth M Robinson2, Wayne S Cutfield1. 1Peds, Univ of
Auckland, Auckland, New Zealand; 2Community Hlth, Univ of Auckland, Auckland, New Zealand.

Aims: To establish whether premature children have metabolic abnormalities during childhood similar to those
observed in term small for gestational age (SGA) children.
Method: 45 prepubertal premature children (gestation 28.3±2.8 weeks, 8 SGA and 36 AGA) and 19 normal weight
prepubertal term controls aged 5-10 years underwent a modified intravenous glucose tolerance test and had insulin
sensitivity assessed using Bergman’s minimal model software. A subgroup of children (17 premature AGA, 7
premature SGA, 11 controls) had IGF-1, IGF2, IGFBP1, IGFBP3 and leptin levels taken at baseline (0900).
Results: Premature children were insulin resistant compared to term controls (Si=15.5±8.1 versus 31.6±22.4,
p<0.0001). Compared to premature AGA subjects being SGA did not modify insulin sensitivity further. In the
premature cohort neither prenatal nor postnatal dexamethasone use affected insulin sensitivity. However insulin
sensitivity was increased in children born to mothers with preeclampsia (13.1±7.2 versus 15.2±6.3, p=0.03). No
effect of macronutients in neonatal period was observed although all the children received inadequate early protein
intake and later elevated fat intake.
Premature infants were shorter than their parents by 0.6±1.1 SDS. They had lower IGF1 (67.1±47.8 mg/l) and
IGFBP3 (1092±245 mg/l) levels compared to similar sized controls (117±49 mg/l and 2340±120 mg/l, both
p<0.0005). IGF1 (r²=0.28, p<0.001) and IGFBP3 (r²=38, p<0.0001) levels were directly correlated and IGFBP1
inversely correlated (r²=58, p<0.0001) to fasting insulin. IGF2 levels were elevated in premature infants (569±143
versus 297±67 mg/l, p<0.0001). The IGF2 levels were directly correlated with both BMISDS (r²=0.22, p<0.05) and
leptin levels (r²=0.31, p<0.01) in premature subjects.
Conclusions: Prematurity is associated with reduced insulin sensitivity independent of growth restriction prior to
birth and thus subjects born prematurely may be at increased risk of adult diseases associated with insulin resistance.
The cause of the prematurity as well as post natal care may modify later insulin sensitivity. Prematurity is also
associated with a reduction in genetic height potential and a hormonal profile consistent with partial growth
hormone insensitivity.

CLINICAL ORAL: Pediatric Endocrinology: Fat & Insulin Sensitivity (11:15 AM - 12:45 PM)

Presentation Date: 6/22/2003, Time: 12:15 PM; Location: Ballroom A




                                                         145
                                   Pediatric Endocrinology: Fat & Insulin Sensitivity

OR42-5 News Summary
Premature children have metabolic abnormalities that may lead to adult diseases

Children born prematurely appear to have metabolic abnormalities, including insulin resistance,
that reduce height and can predispose them to obesity and later adult diseases, according to a
new study being presented on Sunday, June 22, at The Endocrine Society’s 85th Annual Meeting
in Philadelphia.

Low birth weight has been associated with a range of adult diseases, including type-2 diabetes,
hypertension, strokes, heart disease and obesity. These adult diseases have insulin resistance as
an early metabolic abnormality, one that is found often before any other disease develops.

Researchers have previously shown that healthy, growth-retarded children born at term have
insulin resistance by the time they are between 5–10 years old. Most low birth weight subjects
are predominantly born at term and growth retarded, and it is still unclear whether being born
prematurely at the same or lower birth weight – and not growth retarded – also increases the risk
of later adult diseases. As extremely premature infants have only been surviving in substantial
numbers for two to three decades, data on the incidence of these diseases is not yet available.
With the survival of premature children now comprising nearly 5 percent of all births, the impact
of an increase in adult disease has important implications.

To further investigate this issue, researchers in Aukland, New Zealand, led by Dr. Paul Hofman,
investigated a group of 47 prematurely born prepubertal children (less than 32 weeks gestation,
5–10 years old). There were 36 children with appropriate birth weights for gestational age and
11 low birth weight for gestational age who were compared to 20 healthy prepubertal normal
birth weight children of the same age. Insulin sensitivity was measured.

The results demonstrated that all premature children irrespective of whether they were growth
retarded at birth or not were insulin resistant by ages 5–10 years old. However, children born to
mothers with pregnancy-induced high blood pressure – called preeclampsia – were relatively
protected from insulin resistance.

Premature children also have compromised height compared to what would be expected from
their parents heights. This effect was greater in those who were low birth weight for gestational
age. Hormones that play a role in determining height were found to be low in these prematurely
born children. As these hormones are regulated by growth hormone, it is possible these children
also have partial growth hormone resistance.

Finally, the researchers found another hormone, IGF2, that was elevated in premature children.
IGF2 was associated with an indirect but validated measure of fat mass, which may lead to an
increased risk of obesity later in life. Researchers note that this needs further study.

                                              # # #


                                               146
                                         Pediatric Endocrinology: Fat & Insulin Sensitivity
OR42-3
Associations between a Polymorphism in the 11 beta Hydroxysteroid Dehydrogenase Type I Gene and Body
Composition.
Liat Yaniv2, Ningping Feng1, Nancy Sebring1, Zeev Hochberg2, Jack Yanovski*1. 1Unit on Growth and Obesity,
Natl Insts of Hlth, Bethesda, MD; 2Ped Endocrinology, Meyer Children’s Hosp, Haifa, Israel.

It has been hypothesized that increased concentrations of glucocorticoid due to increased conversion of the inactive
glucocorticoid cortisone to its active metabolite, cortisol by 11 beta hydroxysteroid dehydrogenase type I
(11betaHSD-1), may be involved in obesity and its complications, but to date, no polymorphisms significantly
associated with obesity or its complications have been found in the 11betaHSD -1 gene. We therefore investigated
11betaHSD-1 sequence variants in 103 healthy overweight (BMI > 2 SD) and 160 non-overweight (BMI -2 to +2
SD) children to examine the associations between body composition and 11betaHSD-1 polymorphisms. Only one
sequence variant was identified: 4.3% of children were homozygous and 30.0% heterozygous for an adenine
insertion in intron 3 (ins4436A). By ANCOVA (adjusting for age, sex, race, and height), BMI-SD differed
according to ins4436A genotype (p<0.005), with the greatest BMI-SD for ins4436A homozygotes (mean ± SD,
3.4±3.4, vs. heterozygotes, 0.8±5.5, or wild type, 1.8±7.5). Homozygotes also had greater waist circumference
(90.7 ± 12.7 cm ins/ins, 80.6 ± 12.7 cm WT/ins, 86.3 ± 12.6 cm WT/WT, p<0.05), waist-to-hip ratio (0.90 ± 0.06
ins/ins, 0.84 ± 0.08 WT/ins, 0.86 ± 0.10 WT/WT, p<0.05), and insulin resistance indices (HOMA iri, 4.44 ± 3.45
ins/ins, 2.01 ± 5.53 WT/ins, 2.98 ± 7.46 WT/WT, p<0.05) than heterozygote or wild type children, but no significant
differences in trunk fat by DXA, or in serum lipids. We conclude an intronic 11betaHSD-1 gene polymorphism is
associated with greater body mass, altered body composition, and insulin resistance in children. 11betaHSD-1 may
be one of the genes relevant for pediatric-onset obesity and its complications.
          Supported by the National Institute of Child Health, NIH HD-000641, and by the National Center on
Minority Health and Health Disparities, NIH

CLINICAL ORAL: Pediatric Endocrinology: Fat & Insulin Sensitivity (11:15 AM - 12:45 PM)

Presentation Date: 6/22/2003, Time: 11:45 AM; Location: Ballroom A




                                                       147
                                    Pediatric Endocrinology: Fat & Insulin Sensitivity

OR42-3 News Summary
Children with gene alteration are more prone to obesity and precursors of diabetes

Children with an alteration in a gene related to the steroid hormone cortisol are heavier, have
central obesity and insulin resistance, which makes them prone to develop type-2 diabetes,
according to a new study being presented on Sunday, June 22, at The Endocrine Society’s 85th
Annual Meeting in Philadelphia. Drugs targeting this altered gene structure may be a future
treatment for obesity in this group of children.

It was recently discovered that fat and liver produce the hormone cortisol from an inactive
substance called cortisone. An enzyme in the muscle and fat that is called 11-beta-HSD-1
converts cortisone to cortisol. Cortisol increases fat tissue and particularly affects the intra-
abdominal fat that is linked to the complications of being over weight, such as high blood
pressure, high cholesterol and diabetes. If this enzyme is too active, more cortisol is made
locally, and scientists predict that the excess cortisol causes central obesity – obesity of the
abdomen more than the thighs.

Dr. Jack A. Yanovski and colleagues at the National Institutes of Health in Bethesda, Md., and
Meyer Children’s Hospital in Haifa, Israel, studied the gene for this enzyme in a group of 263
children, including 103 healthy, overweight and 160 non-overweight children. They found that
4.3 percent of the group had a changed structure of the gene – called a polymorphism – and, after
adjusting for age, sex, race and height, this group was heavier, had central obesity and insulin
resistance, which makes them prone to develop type-2 diabetes. Currently, several
pharmaceutical companies are developing drugs would block the enzymatic activity of 11-beta-
HSD-1 to treat obesity. The subgroup, identified in the current study, might be the prime
candidates for such therapy, say researchers.

This study was supported by the National Institute of Child Health & Human Development and
the National Center on Minority Health and Health Disparities.

                                                # # #




                                                 148
Reproduction
                                                                            Reproductive Endocrinology
OR28-1
Genetic Screening of 750 Severely Infertile Men Candidates for Intracytoplasmic Sperm Injection.
Alberto Ferlin*1, Lucia Bartoloni1, Giorgia Rizzo1, Carlo Galeazzi1, Enrico Moro1, Carlo Foresta1. 1Dept Med &
Surg Scis, Ctr Male Gamete Cryopreservation, Univ of Padova, Padova, Italy.

Introduction. Recent reports suggested that children born after intracytoplasmic sperm injection performed for male
factor infertility are at increased risk of congenital malformations and chromosome aberrations. These anomalies
seem to be related to the underlying spermatogenic damage of the father, that frequently is caused by genetic
alterations. Methods. We studied 750 severely oligozoospermic men (sperm count < 5 mil/mL) who were
candidates for intracytoplasmic sperm injection. Genetic tests in these subjects included: 1. analysis of peripheral
blood karyotype; 2. analysis of the Y chromosome long arm for detection of microdeletions in the azoospermia
factors; 3. analysis of mutations in the cystic fibrosis gene; 4. analysis of mutations in the androgen receptor gene; 5.
analysis of sperm chromosome aneuploidies. Results. A total of 104 genetic abnormalities were diagnosed,
corresponding to a frequency of 13.9% (104/750). Chromosomal aberrations were present in 5.6% (42/750) and they
were in most cases alterations of the sex chromosomes. Y chromosome long arm microdeletions were detected in
6.0% (45/750) and most frequently included the azoospermia factor c. Mutations in the cystic fibrosis gene were
diagnosed in 1.2% (9/750), whereas mutations in the androgen receptor gene were present in 1.1% (8/750). Sperm
sex chromosome aneuploidies were increased in men with karyotype anomalies and Y chromosome microdeletions,
and also in subjects without genetic abnormalities. Discussion. This study showed that genetic alterations represent
a major cause of severe spermatogenic impairment leading to male infertility. The risk of transmission of
chromosomal and genetic diseases by intracytoplasmic sperm injection is therefore very high. Genetic tests and
genetic counselling are highly recommended in oligozoospermic men candidates for in vitro fertilisation techniques,
to avoid transmission, persistence, or even an increase of genetic defects in future generations.

CLINICAL ORAL: Reproductive Endocrinology (1:00 PM - 2:30 PM)

Presentation Date: 6/21/2003, Time: 1:00 PM; Location: 113 B




                                                          149
                                                              Reproductive Endocrinology
OR28-1 News Summary
Men with severe infertility likely to have genetic defects; common in vitro procedure may
pass bad genes to offspring

Genetic alterations are an important cause of severe male infertility and, therefore, genetic tests
should always be performed to have a correct diagnosis, according to a new study being
presented on Saturday, June 21, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.
In addition, the study found that the risk of transmission of chromosomal and genetic diseases by
an in vitro fertilization technique is very high, making genetic counseling a necessary aspect of a
patient’s consideration for this procedure.

Severe oligozoospermia – a form of male infertility – is the result of a significant reduction of
sperm production. Men with this condition are candidates to undergo in vitro fertilizing
techniques, such as intracytoplasmic sperm injection (ICSI). In ICSI, a single sperm is injected
into an egg that has been surgically removed from an ovary. The egg is transferred to an
incubator where fertilization takes place and then the fertilized egg is implanted into the uterus.
Introduced in 1992, ICSI is used worldwide, comprising nearly 50 percent of in vitro techniques.

However, some concerns about the safety of this procedure have been raised, above all because it
bypasses many physiological processes related to fertilization, and few animal studies were
performed before it was introduced in clinical practice. Furthermore, recent reports have
suggested that children born after ICSI might be at increased risk of congenital malformations
and chromosome and genetic alterations. Therefore, if male infertility is caused by genetic
defects, the use of ICSI may allow these anomalies to be transmitted to offspring. Under normal
circumstances, many of these genetic alterations would not be transmitted; however, ICSI
overcomes this natural selection.

During the last five years, researchers at the University of Padova in Padova, Italy, led by Dr.
Alberto Ferlin, performed a genetic screening of 750 infertile men with oligozoospermia,
performing tests for the most common alterations causing severe male infertility: 1) analysis of
karyotype for numerical and structural chromosomal aberrations; 2) analysis of small alterations
(microdeletions) in particular regions, named AZF, of the long arm of the Y chromosome; 3)
analysis of mutations in the cystic fibrosis gene; 4) analysis of mutations in the gene encoding
the receptor for androgens; and 5) analysis of numerical chromosomal alterations in sperm. The
researchers found a high percentage of genetic abnormalities, with a total of 104 men out of 750
with some alterations (13.9 percent), compared to less than 1.5 percent that are normally found in
fertile men. In particular, chromosomal aberrations were present in 5.6 percent, Y chromosome
microdeletions in 6 percent, cystic fibrosis gene mutations in 1.2 percent and androgen receptor
gene mutations in 1.1 percent.

Furthermore, men with karyotype anomalies and Y chromosome microdeletions produced a
higher number of sperm with chromosome aneuploidy, as did men in whom no genetic
abnormality was detected.
                                          # # #


                                               150
                                                                            Reproductive Endocrinology
OR28-2
Short-Term Testosterone Supplementation in Patients with Erectile Dysfunction: Evidence for Improvements
in Cavernous Arteries Vasodilation and Sildenafil Response.
Antonio Aversa*1, Andrea M Isidori1, Massimiliano Caprio2, Giovanni Spera1, Andrea Lenzi2, Emanuela Greco1,
Andrea Fabbri2. 1Med Pathophysiology, Univ ‘La Sapienza’Rome, Italy; 2Endocrinology Unit - Dept of Intern Med,
Univ ‘Tor Vergata’Rome, Italy.

We have recently shown that in men with erectile dysfunction (ED) free testosterone (FT) directly correlates with
penile arterial inflow. This led us to further investigate the effect/s of androgen administration on cavernous arteries
in patients (pts) failing sildenafil treatment. In a prospective randomised placebo-controlled pilot study we have
investigated 20 pts with arteriogenic ED as evaluated by dynamic Color-Duplex Ultrasound (D-CDU) studies,
normal sexual desire but testosterone (T) and FT in the lower quartile of normal range (low-normal), not responding
to sildenafil treatment (100 mg) on six consecutive attempts. All pts had D-CDU, hormonal (LH, PSA, total and free
testosterone, SHBG, estradiol), biochemical (hematocrit, LDL- and HDL-cholesterol, triglycerides), and sexual
evaluations [International Index of Erectile Function (IIEF)] before and after one month of therapy with transdermal
testosterone (5 mg/day, N=10) or placebo along with sildenafil treatment on demand. Measurement of flow
parameters by D-CDU on cavernous arteries was the primary endpoint of the study. Improvement of erection was
assessed using the IIEF questionnaire and the Global Assessment Question (GAQ). One month treatment with
transdermal testosterone led to a significant increase in T and FT levels (23.7 ± 3.3 SD vs. 12.8 ± 2.1 nmol/L and
473 ± 40.2 vs 260 ± 18.1 pmol/L, p < 0.01 respectively). In addition testosterone administration induced both a
significant increase in arterial inflow to cavernous arteries measured by D-CDU (32 ± 3.6 vs 25.2 ± 4 cm/sec, p <
0.05) and smooth muscle cell relaxation (resistive index 0.85 ± 0.16 vs 0.80 ± 0.12, p < 0.05), with no adverse
effects. Also, a significant improvement in erectile function domain score at IIEF was found in the androgen but not
in the placebo treated pts (21.8 ± 2.1 vs 14.4 ± 3.1 p < 0.05) which was associated with significant changes in the
GAQ score (80% vs. 10%, p < 0.01). Short-term T administration in ED pts with low-normal androgen levels
increases androgen levels and improves the erectile response to sildenafil by increasing vascular smooth muscle
cells relaxation and arterial inflow into the penis during sexual stimulation.
          This work was supported by a grant of the Ministero Universita e della Ricerca Scientifica e Tecnologica
(MURST-COFIN 40%, 2002).

CLINICAL ORAL: Reproductive Endocrinology (1:00 PM - 2:30 PM)

Presentation Date: 6/21/2003, Time: 1:15 PM; Location: 113 B




                                                         151
                                                                 Reproductive Endocrinology

OR28-2 News Summary
Testosterone treatment jump starts effects of Viagra in certain men

Testosterone treatment in testosterone-deficient men improves sexual function, including arterial
flow to the penis, allowing these men who previously did not respond to Viagra to overcome
erection problems, according to a study being presented on Saturday, June 21, at The Endocrine
Society’s 85th Annual Meeting in Philadelphia.

Testosterone is the main androgen, or male sex hormone, involved in the regulation of male
sexual function and exerts its action at three different levels: the brain, the spinal cord and inside
the penis, having a major role in the control of penile erection. Erectile dysfunction (ED) due to
hypogonadism – decreased or absent secretion of hormones from the testes, such as total
testosterone levels less than 2.5 ng/ml, which is present in about 30 percent of the male elderly
population – is thought to be responsive to androgen replacement therapy (ART).

However, researchers say that the role of ART in mild hypogonadic patients with ED has to be re-
evaluated in the presence or absence of drugs like Viagra (sildenafil). The difficulties to perform
sex lead to a reduction attempts to have sex, a fall in sexual interest and a reduction of circulating
androgen levels, which perpetuates the cycle. Overall, these factors show that testosterone is
involved in maintenance of sexual activity in many capacities, and adequate levels are necessary
the keep intact the molecular machinery of penile erection.

In this short-term study, Dr. Antonio Aversa and his colleagues in Rome recruited 20 patients
(average age of 56) with ED due to arterial blood flow problems, normal low testosterone levels
and an insufficient response to Viagra. Half of the participants used testosterone patches for one
month, and the other half a placebo patch. The results showed that testosterone improved
erections and the overall efficacy of Viagra compared with placebo treatment.

Accordingly, arterial flow to the penis improved significantly, thus suggesting a direct effect of
testosterone on penile vasculature. No side effects were reported by the patients throughout the
study period.

This paper represents the first demonstration, note the researchers, that testosterone positively
affects penile arterial function in men. This opens new perspectives in the management of aging
men, they add, in which partial or total androgen deficiency is very common. The dilemma to
treat or not to the deficiency with androgen substitution remains open to discussion.

This work was supported by a grant of the Ministero Universita e della Ricerca Scientifica e
Technologica.

                                                # # #




                                                 152
                                                                                    Female Reproduction
P2-190
Endothelial Dysfunction in Polycystic Ovary Syndrome.
Evantia Diamanti-Kandarakis*1, Krystalenia Alexandraki1, Athanasios Protogerou2, Pantelis Chatzismalis1, Thenia
Koutsoumpa1, John Lekakis2. 1Endocrine Sect, First Dept of Med, Laiko Hosp, Med Sch, Univ of Athens, Athens,
Greece; 2Dept of Clin.Therapeutic, Alexandra Univ Hosp, Athens, Greece.

Introduction: Diverging conclusions exists in polycystic ovary syndrome (PCOS) regarding endothelial dysfunction.
Purpose of the study was to investigate endothelial function in PCOS.
Methods: We studied 19patients (aged: 25.7±6.0) meeting the diagnostic criteria for PCOS of 1990 (NIH consensus)
and 14 normal women (aged: 27.1±3.2). None of the subjects we studied had diabetes mellitus, dyslipidemia or
hypertension. Three women in PCOS group and seven women in control group were smokers. Endothelial function
was assessed by flow-mediated dilatation (FMD, %) in the brachial artery. Serum total testosterone (T, ng/dl), serum
steroid hormone binding globulin (SHBG, nmol/L), serum fasting insulin (INS, IU/L), serum fasting glucose
(GLU, mg/dl), and the ratio GLU/INS were measured in all subjects.
Results: FMD was statistically lower in PCOS (3.5±0.8 vs 6.6±0.6, p=0.009) while endothelial-independent
dilatation assessed by nitrate induced dilatation (NID) was similar within two groups (14.6±1.6 vs 14.0±1.9, ns).
The two groups differed in T (95.4±9.0 vs 44.0±5.1, p=0.0001), in SHBG (27.7±3.1 vs 48.5±5.5, p=0.004). INS and
GLU were approaching statistical significance within two groups (15.7±5.2 vs 5.7±0.3, p=0.07 and 79.7±3.1 vs
72.7±1.5, p=0.07) respectively. The ratio GLU/INS did not differ statistically within two groups (11.0±1.7 vs
13.2±0.8, ns). Body mass index (BMI, Kg/m²) was statistically higher in PCOS (28.6±1.9 vs 21.0±0.6). In subgroup
analysis of subjects with BMI<27 differences in FMD, T and SHBG remaimed statistically significant within two
groups. In our population FMD was related to BMI (p=0.003, r=-0.508), T (p=0.040, r=-0.371), SHBG (p=0.016,
r=0.431), INS (p=0.011, r=-0,451), GLU/INS (p=0.017, r=0.425). In multiple regression analysis GLU/INS and T
were independent predictors of endothelial dysfunction (p=0.09, b=0.461 and p=0.034, b=-0.362, respectively).
Conclusion: In the present study endothelial function was found impaired in PCOS when compared to women with
normal menstrual cycle and normal androgens levels. It seems that hyperandrogenism and insulin resistance
propably participate in the mechanisms responsible for endothelial dysfunction in PCOS.

CLINICAL POSTER: Female Reproduction (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                       153
                                                                        Female Reproduction
P2-190 News Summary
Blood vessel function is impaired, leading to heart disease, in women with common
endocrine problem

Women with a common endocrine disorder, called polycystic ovary syndrome, have impaired
function of their blood vessels, which is a very early sign of atherosclerotic disease, according to
a new study being presented on Friday, June 20, at The Endocrine Society’s 85th Annual Meeting
in Philadelphia.

PCOS is the most common endocrine problem among women of reproductive age, affecting 4
percent to 6 percent of this group. Women with PCOS have excess facial and body hair
(hirsutism), acne, menstrual abnormalities and fertility problems. In addition, it has been found
that they are at increased risk for heart problems later in life because of metabolic aberrations
like insulin resistance, glucose intolerance, dyslipidemia, coagulation problems and elevated
cardiovascular risk factors in the blood (endothelin-1, CRP and adhesion molecules).

For the last 15 years, researchers in Athens, Greece, have been studying women with this
syndrome. In the current study, led by Dr. Evanthia Diamanti-Kandarakis, they studied 19
PCOS patients and 14 normal young women with a very simple, non invasive method – flow
mediated dilatation (FMD) by ultrasonography. This imaging test, which is highly reproducible
with no side effects – allows assessment of blood vessel, or vascular, function. The researchers
found that FMD was related to obesity, to androgens (male sex hormones) and insulin (a
chemical that lowers blood sugar) and to an index of insulin resistance, which is a basic
constituent of metabolic syndrome – a cluster of cardiovascular risk factors.

Women who show similar symptoms to this high-risk group of women with PCOS with early
cardiovascular manifestation should be tested, say the researchers, because lifestyle
modifications and certain medications could prevent or ameliorate the abnormalities. In
addition, the test is simple, with no side effects. Otherwise, they may be candidates for diabetes
and heart problems later in life.

                                              # # #




                                                154
                                                                                   Female Reproduction
P2-218
A Comparative Study of the Pharmacokinetics of Testosterone (T) Following Administration of a Metered-
Dose Transdermal Spray (MDTS®) to the Abdomen or Forearm in Healthy Postmenopausal Women with
Low Serum Testosterone.
Andrew J Humberstone*1, Allan M Evans2, Susan R Davis3. 1Acrux Ltd, West Melbourne, VIC, Australia; 2Sch of
Pharmaceutical, Molec and Biomed Scis, Univ of South Australia, Adelaide, SA, Australia; 3The Jean Hailes Fdn,
Clayton, VIC, Australia.

Background: Testosterone MDTS is being developed for the treatment of women with low libido associated with
low serum T levels. The MDTS delivers an accurate small volume of a fast drying solution containing T and
penetration enhancer to the skin, forming a depot from which T is slowly absorbed. The applied dose for this study
was chosen in a pilot study in surgically postmenopausal women(1).
Study population: Healthy postmenopausal women with low serum total T stabilized on transdermal estrogen
therapy.
Aims: To confirm the daily dose required to increase average serum free testosterone levels into the mid-to-high
normal range for premenopausal women (approx 5 pg/mL for the assay used), and to compare the steady-state
pharmacokinetics of free and total T after daily administration of the Testosterone MDTS to the abdomen or the
forearm.
Methods: Subjects were administered 2 sprays of the MDTS daily for 5 days on two occasions either to the
abdomen or the forearm in a randomized cross-over design. Intensive blood sampling was performed on day 5 and
samples were analyzed for serum free and total T concentrations to determine pharmacokinetic parameters. Total T
was measured by chromatography and RIA and free T was measured using equilibrium dialysis by Esoterix
Endocrinology, CA, USA.
Results: 14 women, mean age 60 years (range 50–77), completed the study. 11 women were naturally
postmenopausal and 3 had prior bilateral ovariectomy. Mean (±SD) average and maximum serum concentrations
(Cavg and Cmax) and mean baseline concentrations for free and total T are shown in the table below.
                           baseline      Cavg       Cmax       Normal*
free T (pg/mL) abdomen 1.7 ± 0.6 5.3 ± 1.9         7.8 ± 3.2 1.1–6.3
                 forearm 1.7 ± 0.5 7.4 ± 3.3 13.0 ± 7.8
total T (ng/dL) abdomen 20.3 ± 7.0 57.8 ± 18.2 81.1 ± 29.9 10–55
                 forearm 20.1 ± 6.3 77.6 ± 34.2 130.5 ± 90.4
* Normal range for premenopausal women
Conclusions: Application of a daily dose of Testosterone MDTS for 5 days to the abdomen resulted in increased
serum concentrations of both free and total T. Mean Cavg for free serum T was increased into the mid-to-high
normal range for premenopausal women. Average serum T concentrations were significantly higher after
administration to the forearm compared to the abdomen.
          Reference: 1. Humberstone, AJ and Williamson, MM. Elevation of serum testosterone levels in
oophorectomized women following application of a novel Testosterone MDTS. Presented at the NAMS Annual
Meeting, Chicago, Oct 2002.

CLINICAL POSTER: Female Reproduction (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                       155
                                                                        Female Reproduction
P2-218 News Summary
Testosterone spray shown to increase testosterone to healthy levels in certain women

An investigational testosterone spray for women is found to increase testosterone levels into a
healthy range when two sprays are applied daily to the abdomen for five days, according to a
new study being presented on Friday, June 20, at The Endocrine Society’s 85th Annual Meeting
in Philadelphia. These new data will be the basis of a larger study to compare the clinical effect
and safety of this dose to a placebo in women who have lowered libido and low testosterone.

Testosterone levels decrease with age in otherwise healthy women. There is increasing
recognition that a cluster of symptoms including low libido, fatigue and reduced sense of well-
being may be related to low testosterone levels – also called androgen insufficiency – in some
women. Several researchers have shown that these symptoms can be improved by the
administration of testosterone. However, establishing a minimum effective and safe dose has
been limited by lack of availability of low-dose testosterone formulations suitable for use in
women.

Dr. Andrew Humberstone and colleagues in Adelaide and Clayton, Australia, conducted a study
as part of the initial testing of a novel mode of administering testosterone to women. The
investigational product is fast-drying testosterone spray. When applied to the skin once a day,
the spray forms an invisible patch within the skin from which the testosterone is absorbed slowly
into the circulation.

Fourteen healthy postmenopausal women with low testosterone levels took part in the study.
The aim of the study was to confirm the number of sprays required to increase testosterone levels
into the mid-to-high range for healthy young women. The researchers also compared two
different application sites: the abdomen and the forearm.

The results confirmed that two sprays applied daily to the abdomen for five days increased
testosterone levels into the required range. They also showed that absorption was higher but
more variable when applied to the forearm.

One of the authors, Dr. Susan Davis is a leading researcher in the area of testosterone therapy in
women. Most recently, her research group has shown that testosterone therapy, using a cream,
significantly improved libido in premenopausal women, 30–45 years old.

This study was supported by Acrux Ltd of Australian.

                                              # # #




                                               156
                                                                                       Female Reproduction
P2-214
Estrogen Replacement Reduces Errors in Verbal Perseveration, but Does Not Enhance Other Cognitive
Domains in Menopausal Women: A Randomized, Double-Blind, Placebo-Controlled Study.
Hadine Joffe*1,2,3, Deborah Yurgulen-Todd3,4, Janet E Hall3,5, Ingrid A Sarmiento1, Staci A Gruber3,4, Lee S
Cohen2,3, Kathryn A Martin3,5. 1Women’s Ctr for Behav Endocrinology, McLean Hosp, Belmont, MA; 2Perinatal
and Reprod Psychiatry Clin Res Prog, Massachusetts Gen Hosp, Boston, MA; 3Harvard Med Sch, Boston, MA;
4
  Cognitive and Neuroimaging Ctr, McLean Hosp, Belmont, MA; 5Reprod Endocrine Unit, Dept of Med,
Massachusetts Gen Hosp, Boston, MA.

Background: Previous trials report inconsistent findings about whether estrogen replacement therapy (ERT)
improves cognition in postmenopausal women. Cognitive enhancement with ERT has been attributed to inclusion of
subjects with hot flushes and sleep disruption, but the effect of ERT on cognitive performance has not been
examined specifically in symptomatic menopausal women.
Methods: In a double-blind trial, we randomly assigned 52 menopausal women to receive estradiol 0.05 mg/day
transdermal patch (26 patients) or a placebo patch (26 patients) for 12 weeks. All subjects underwent neurocognitive
testing using an abbreviated battery that included measures of attention, verbal, and visual memory at both baseline
and study-end. The primary end-point was change in verbal recall performance on the California Verbal Learning
Test. Vasomotor symptoms and sleep quality during the month prior to each study visit were recorded.
Results: Subjects were perimenopausal (n=29), postmenopausal (n=21), and hysterectomized (ovaries intact, n=2)
women (ages 51.1 ± 3.7) who had moderate or severe vasomotor symptoms (32.7%), mild symptoms (30.8%), or
were asymptomatic (36.5%). Baseline serum FSH levels were comparable between treatment groups (77.7 ± 46.1
IU/L for ERT group vs. 97.9 ± 58.1 IU/L for placebo group). There was no effect of ERT use on verbal recall (59.2
± 8.3 at baseline to 59.8 ± 9.6 at study-end for ERT-treated vs. 58.0 ± 8.2 at baseline to 60.6 ± 9.4 at study-end for
placebo-treated subjects, p=0.48). However, ERT reduced perseverative errors on CVLT Trials 1- 5 more than
placebo (6.5 ± 5.5 at baseline to 3.7 ± 3.3 at study-end for ERT-treated vs. 6.2 ± 5.4 at baseline to 5.6 ± 3.8 at study-
end for placebo-treated subjects, p=0.03). There was no effect of ERT on tests of other cognitive domains examining
attention, verbal or visual memory. There was significant correlation between improvement in verbal recall and
reduction in vasomotor symptoms (Spearman r=- 0.34, p=0.02), but no correlation with improvement in sleep
(Spearman r<0.01, p=0.95).
Conclusions: While ERT reduced errors in verbal perseveration, there was no global benefit of ERT for cognitive
performance in menopausal women, despite a predominately symptomatic menopausal study population. Our
finding of reduced errors in verbal perseveration with ERT treatment requires further investigation.
          Supported by Pfizer/Society for the Advancement of Women's Health Research Scholars Grant

CLINICAL POSTER: Female Reproduction (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                          157
                                                                       Female Reproduction
P2-214 News Summary
Estrogen therapy improves cognitive processes in verbal learning and memory

Estrogen replacement therapy (ERT) selectively improves specific cognitive processes involved
in verbal learning and memory during verbal recall in perimenopausal and recently
postmenopausal women, according to a new study being presented on Friday, June 20, at The
Endocrine Society’s 85th Annual Meeting in Philadelphia. This cognitive benefit is not
explained by improvement in hot flushes or sleep, suggesting that ERT enhances the efficiency
of verbal information processing.

There is much debate about the benefits and risks of hormone replacement therapy. Results of
clinical trials examining cognitive effects of ERT have been mixed, with some, but not all,
suggesting that there are specific processes enhanced by ERT use. However, it has also been
suggested that improvement in cognitive processes is due to the inclusion of recently
postmenopausal women whose hot flushes and insomnia resolve with ERT.

However, this question has not been addressed by studying perimenopausal – the period around
the onset of menopause – and recently postmenopausal women, who are most likely to have hot
flushes and associated sleep disruption. For this reason, Dr. Hadine Joffe and colleagues in
Boston and Belmont, Mass., designed a randomized, double-blind, placebo-controlled trial to
determine whether ERT influences selective cognitive processes in these women.

Perimenopausal and recently postmenopausal women with complaints of cognitive changes
during menopause transition were recruited from the community. The age of the 52 study
participants ranged from 4560 years old. Participants were assigned randomly to an estrogen
patch (n=26) or an identical placebo patch (n=26). Before starting treatment, all participants
completed an assessment of the short-term verbal and visual memory and learning skills using
standardized methods of measuring cognitive function. At the end of the 12th week of treatment,
the same tests were repeated so that changes in performance on these tests could be determined.

There was no effect of ERT use on overall verbal recall. However, ERT reduced perseverative
errors during verbal recall more than placebo, reflecting a 43 percent reduction of perseverative
errors in ERT-treated and a 9 percent reduction in placebo-treated subjects. Perseveration is the
uncontrollable repetition of a previously appropriate or correct response, even though the
repeated response has since become inappropriate or incorrect. Although hot flushes improved
more with ERT than with placebo treatment, the benefits of ERT for hot flushes and associated
sleep disruption did not explain the cognitive benefits observed. These results, say researchers,
provide important information for risk-benefit decisions about ERT use in this population.

This work was supported by the Pfizer/Society for the Advancement of Women’s Health
Research Scholars Grant.

                                              # # #



                                               158
                                                                                      Female Reproduction
P2-208
Estrogen Bio-Activity in Fo-Ti and Other Herbs Used for Their Estrogen-Like Effects as Determined by a
Recombinant Cell Bioassay.
Karen O Klein*1, Mona Janfaza1, Jeffrey A Wong1, R Jeffrey Chang1. 1Peds and Reprod Med, Univ of California -
San Diego, San Diego, CA.

One of the most important issues in women’s health concerns the risks and benefits of estrogen replacement therapy.
However, continual uncertainty and lack of consensus of standard ERT has driven many women to seek alternative
sources of estrogen, including herbal remedies. To determine the presence and extent of estrogen in herb extracts,
we adapted a recombinant cell bioassay for estradiol to quantify estrogen bio-activity in selected herbs. Using this in
vitro system, estrogen bio-activity was measured in red clover, dong quai, black cohosh, soy, licorice, chaste tree
berry, fo-ti, and hops. We found that soy, red clover, licorice, and hops have a large amount of measurable estrogen
bio-activity which is consistent with previous reports using other methods. There was a surprisingly high degree of
estrogen activity in extracts of Fo-Ti, which has not been previously reported. Chaste Tree Berry, Black Cohosh, and
Dong Quai did not have measurable activity. We also discovered that removal of a glycone group from soy (SoyA)
significantly increases its estrogen bio-activity.
               Estrogen Bio-activity in Herbs
   Herb Mean (pg/mL) per mcg herb Significance
   Chaste            Not measurable
  Cohosh             Not measurable
Dong Quai            Not measurable
   Fo-Ti                 111 ± 15                   b
   Clover                 85 ± 39                   b
   Hops                 0.2 ± 0.06                  a
  Licorice                 20 ± 3                  a,b
    Soy                   46 ± 13                  a,b
   SoyA                 396 ± 106                   c
mean ± SD, Values with different letters are significantly different from each other.
We conclude that this recombinant cell bioassay for estradiol can be used to measure bio-activity in herbal extracts.
The preparations of soy, clover, licorice and fo-ti studied have estradiol activity in the range of 0.2 – 400 pg/mL
estradiol equivalents per mcg of herb, which is approximately 1/300 th the activity of 17B-estradiol itself. Clinical
studies are underway to determine the estrogen bio-activity present in women using dietary supplements containing
these herbs.

CLINICAL POSTER: Female Reproduction (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                         159
                                                                        Female Reproduction
P2-208 News Summary
Estrogen found in Chinese healing herb, among other natural products

Researchers developed a method to detect naturally occurring estrogen and found it in herbs like
hops, red clover, soy and licorice as well as in the ancient Chinese healing herb fo-ti, which is a
first-time finding, according to a new study being presented on Friday, June 20, at The Endocrine
Society’s 85th Annual Meeting in Philadelphia. This study shows the potential for estrogen’s
effects in natural products, say the researchers, and highlights the need for further study of these
products, both to determine whether products have the effects that are claimed and to study the
potential risks associated with these products.

One of the most important issues in women’s health concerns the risks and benefits of hormone
treatments used during menopause. Menopause is the time during which female hormones,
primarily estrogen, decrease in a woman’s body. This change can cause hot flashes – a short
interval of feeling warm, vaginal dryness, decreased sexual desire, thinning of the skin and
decreased thickness of bone. Some women are prescribed estrogen pills – called estrogen
replacement therapy (ERT) – by their physicians. Other women seek natural alternatives to
ERT. The actual estrogen action in natural products is not well known, however.

Dr. Karen O. Klein and colleagues at the University of California in San Diego developed and
used a method to measure estrogen activity in herb products. They found estrogen activity in
some herbs as others have also shown, including hops, red clover, soy and licorice. They
discovered estrogen activity in fo-ti, which has not been reported previously. They also
discovered that soy products can be processed in a way to increase the amount of estrogen
activity. They did not find estrogen activity in chaste tree berry, balck cohosh or dong quai. The
researchers are now ready to study the estrogen effect of these herb products in women using
them. The findings of the future studies will help women know what effects herb products are
having on their bodies.

This study was done in the laboratory and not in women. The system used to measure estrogen
activity involved exposing yeast, designed to be sensitive to estrogen, to liquid extracts from the
different herb plants. The yeast grow and respond to any estrogen-like substances in the herbs,
such that the amount of estrogen activity can be measured.

This study was supported by Natural Alternatives International, Inc.

                                              # # #




                                                160
                                                                                     Female Reproduction
P2-195
The Association of Endogenous Androgen Levels with Metabolic Risk Factors for Coronary Heart Disease in
Postmenopausal Women.
Hyesoo Lowe*1, Laurie Feldman2, Mihaela Aslan2, Teresa Caulin-Glaser3, Christopher Howes4, Barbara Gulanski5.
1
  Intern Med, Yale Univ Sch of Med, New Haven, CT; 2Intern Med, Ethel F. Donaghue Women’s Hlth Investigator
Prog, New Haven, CT; 3Intern Med/Cardiol, McConnell Heart Hlth Ctr, Columbus, OH; 4Intern Med/Cardiol, Yale
Univ Sch of Med, New Haven, CT; 5Intern Med/Endocrinology, Yale Univ Sch of Med, New Haven, CT.

Although much attention has been focused on the relationship between endogenous androgens and metabolic risk
factors for coronary heart disease (CHD) in women with PCOS, less data are available regarding this association in
postmenopausal women. It has been postulated that increased androgenicity may contribute to the increased CHD
risk seen after menopause via its relationship to lipid levels, vascular inflammatory markers, and insulin resistance.
We measured fasting serum lipids (total cholesterol, HDL, LDL and triglycerides), vascular inflammatory markers
(C-reactive protein [CRP], interleukin-6 [IL-6]), insulin resistance (as measured by homeostasis model assessment,
HOMA), serum androgens, and measures of obesity (BMI and waist circumference) in healthy postmenopausal
women, none of whom were taking exogenous gonadal steroids. The primary androgen for the analyses was the free
androgen index (FAI) calculated as total testosterone (nmol/L)/SHBG (nmol/L) x100.
Thirty-one women (mean age 56.8 years, range: 44-64) are included in these analyses. The mean BMI was 27.6
(range: 20-40.2). The majority (87%) were caucasian. There was a trend toward a higher mean FAI for obese
women (BMI 30) compared to overweight (BMI 25-29.9) and lean (BMI <25) women (21.9 vs 8.16 vs 7.75,
respectively).
In bivariate analyses (using Spearman correlation coefficients), FAI was significantly correlated with CRP (r=0.61,
p=0.001), IL-6 (r=0.44, p=0.046), LDL (r=0.62; p=0.008), triglycerides (r=0.40, p=0.05), and showed a trend with
HDL (r=-0.38, p=0.06). Preliminary analyses also revealed a significant correlation between FAI and HOMA
(r=0.50; p=0.02). After adjustment for BMI, FAI was no longer significantly associated with CRP or IL-6. However,
FAI remained significantly and positively associated with LDL (p=0.001) and inversely related to HDL (p=0.053).
A similar pattern was observed after adjustment for waist circumference.
In conclusion, endogenous androgens as measured by FAI were associated with a dyslipidemic profile even after
adjustment for anthropometric measures of adiposity. However, FAI was no longer associated with CRP and IL-6
after controlling for fat mass. These data suggest that higher endogenous androgen levels are predictive of an
adverse lipid profile in healthy postmenopausal women, and that this finding is independent of obesity.
          Supported by Ethel F. Donaghue Women’s Health Investigator Program at Yale
          Yale-New Haven Hospital General Clinical Research Center

CLINICAL POSTER: Female Reproduction (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                        161
                                                                        Female Reproduction
P2-195 News Summary
High levels of testosterone in postmenopausal women may be culprit of increased heart
disease risk

Higher levels of androgens – male sex hormones like testosterone – in postmenopausal women
are associated with an unfavorable lipid profile and insulin resistance, which are known risk
factors for cardiovascular disease, according to a new study being presented on Friday, June 20,
at The Endocrine Society’s 85th Annual Meeting in Philadelphia. The results of this preliminary
study, say researchers, suggest that androgens, or a disruption of the estrogen–androgen balance,
may play a critical role in the development of cardiovascular disease in postmenopausal women.

For women in the United States, the risk of heart disease is increased after the transition to
menopause and is the primary cause of death. This risk is further increased among diabetic
women. The increased incidence of heart disease seen after menopause, when estrogen levels
decrease, has been largely attributed to the loss of estrogen’s beneficial effects. However, the
cardiovascular benefits of estrogen have recently been called into question, with little benefit
seen in studies of hormone replacement therapy (HRT).

Androgens that women produce may play a significant role in cardiovascular risk, particularly in
women with diabetes. Much of the research about androgens and heart disease risk has focused
on premenopausal women with the polycystic ovary syndrome (PCOS), who have high androgen
levels, obesity and insulin resistance, a precursor to diabetes. Women with PCOS have
been shown to have a significantly increased risk for heart disease, suggesting a
relationship between insulin resistance, high androgen levels and cardiovascular disease. Some
researchers have suggested that increased androgens may contribute to cardiovascular disease
through the relationship to lipid levels, blood vessel inflammation and insulin resistance.
However, there had been little data about this association in postmenopausal women.

For this reason, Dr. Hyesoo Lowe and colleagues in New Haven, Conn., and Columbus, Ohio,
designed a study that included 31 healthy postmenopausal women, 44–64 years old, and they
measured obesity, low-density lipoprotein (LDL; “bad”) and high-density lipoprotein (HDL;
“good”) cholesterol, insulin resistance, markers of blood vessel inflammation (C-reactive protein
[CRP] and interleukin-6 [IL-6]) and androgens. The participants, mostly Caucasian, were not on
HRT and were overweight (average body mass index [BMI] was 27.6; range of 20–40.2).

They found that higher androgen levels are significantly correlated with cardiac risk factors,
including LDL cholesterol, triglycerides and insulin resistance. Women with obesity (BMI≥30)
had significantly higher androgen levels than did lean women (BMI<25). Even after adjusting
for obesity by accounting for BMI and waist circumference, the relationship between androgens
and cardiac risk factors remained significant.

The research was supported by the Yale-New Haven Hospital General Clinical Research Center
and the Yale University Ethel F. Donaghue Women’s Health Investigator Program.
                                           # # #


                                               162
                                                                                     Female Reproduction
P2-215
Healthy Young Women Compared to Men Are More Resilient to Sleep Loss and Sleep Disturbance Than
Age-Matched Men: Potential Protective Role of Estrogen.
Alexandros N Vgontzas*1, Emmanouil Zoumakis2, Edward O Bixler1, Hung-Mo Lin3, Heather Follett1, George P
Chrousos2. 1Psychiatry, Sleep Res & Treatment Ctr, Penn State Univ Coll of Med, Hershey, PA; 2Ped and Reprod
Endocrinology Br, NIH, Bethesda, MD; 3Hlth Evaluation Scis, Penn State Univ Coll of Med, Hershey, PA.

Objectives: Total sleep deprivation or even modest sleep restriction to 6 hours per night is associated with increased
daytime sleepiness, decreased performance and increased secretion of pro-inflammatory cytokines. The goal of this
study was to assess whether pre-menopausal women have better quality responses to sleep loss and sleep
disturbance than age-matched men.
Methods: Twenty-five young healthy normal sleepers, 12 men and 13 women, were recorded in the sleep laboratory
for 12 consecutive nights (4 baseline nights during which subjects were allowed to sleep for 8 hours followed by 1
week of sleep restriction to 6 hours). At baseline and following 1 week of sleep restriction we obtained measures of
daytime sleepiness (multiple sleep latency test) (performance psychomotor vigilance test) and serial twenty-four
hour plasma measures of IL-6, TNF, and cortisol.
Results: Following one week of sleep restriction, men but not women showed a significant elevation of plasma
TNF levels (P < 0.05). Also cortisol secretion in the morning was significantly lowered only in men. As
anticipated after one week of sleep restriction subjects demonstrated shorter sleep latencies (SL), increased
percentage of sleep time (% ST), more SWS (deep sleep) and less stage 1 sleep (light sleep) (all P < 0.05). However,
the described homeostatic responses of SL, % ST and % SWS to sleep restriction were significant in women but not
in men (P < 0.05). Furthermore evaluation of the sleep disturbing effects of blood drawing showed a stronger effect
in men vs. women (significantly higher percentage of light sleep (P < 0.05) and similar trends in terms of SL and %
ST).
Conclusions: Pre-menopausal women are more resilient to the effects of sleep loss in terms of hormonal changes
whereas their sleep during sleep restriction becomes more consolidated (stronger homeostatic response).
Furthermore women appear to be more resistant to external sleep disturbance. These results are consistent with
recent data that women’s sleep across all ages is of better quality and quantity (Bixler et al., 2003). This marked
sexual dimorphism in sleep regulation may have been to protect women from the profound demands of infant and
childcare for most of mankind’s history. Furthermore these data collectively, may explain to some extent the
differences between men and women in terms of cardiovascular risks and longevity.
         Supported by National Institutes of Health Grants R01 HL64415, HL40916, and HL51931.

CLINICAL POSTER: Female Reproduction (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                        163
                                                                        Female Reproduction
P2-215 News Summary
Women’s sleep is better in quantity and quality than men’s, may stem from adaptation

Pre-menopausal women are more resilient than men to the effects of sleep loss and more
resistant to stress-related sleep disturbance, according to a study being presented on Friday, June
20, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

Dr. Alexandros Vgontzas and colleagues at the Pennsylvania State University College of
Medicine in Hershey, Pa., and the National Institute of Child Health and Human Development in
Bethesda, Md., recruited 25 healthy young men (n=12) and women (n=13) and asked them to
forego two hours of sleep for one week so that they received only six hours of sleep, instead of
the customary eight hours. The researchers tested the study volunteers for the quantity and
quality of nighttime sleep, daytime alertness and performance and measured their levels of
immune and stress hormones.

Following one week of sleep restriction to six hours per night, men but not women secreted
higher amounts of the immune hormone tumor necrosis factor-α. Also, following that week of
sleep restriction, women’s sleep became deeper compared to men. Furthermore, women’s sleep
was more resistant to sleep disturbance induced by a mild stressor, such as blood drawing.

These results are consistent with recent data that women’s sleep across all ages is of better
quality and quantity (Bixler et al., 2003). The researchers say that this marked sexual
dimorphism in sleep regulation may have been to protect women from the profound demands of
infant and childcare for most of mankind’s history. Furthermore, these data collectively may
explain to some extent the differences between men and women in terms of cardiovascular risks
and longevity.

This study was supported by the National Heart, Lung and Blood Institute.

                                              # # #




                                                164
                                                                                     Female Reproduction II
P3-15
Neonatal Exposure to the Soy Phytoestrogen Genistein Alters Mammary Gland Differentiation and
Development.
Elizabeth Padilla-Banks*1, Wendy N Jefferson1,2, Retha R Newbold1. 1Lab of Molec Toxicology, Natl Inst of Envir
Hlth Scis, NIH/DHHS, Research Triangle Park, NC; 2Dept of Envir and Molec Toxicology, North Carolina State
Univ, Raleigh, NC.

The effect of soy on the breast is controversial. Some reports suggest developmental exposure of rats to genistein
protects against mammary carcinogens later in life; this effect is reported to be correlated with a decrease in
carcinogen-sensitive undifferentiated terminal end buds in the mammary gland. However, in vitro studies suggest
the opposite, i.e., that genistein stimulates breast cancer cells to proliferate. Further, exposure to estrogens, including
the period of fetal development, is a well-known risk factor for the development of breast cancer. Also, numerous
experimental studies involving the mammary gland and the reproductive tract attest to the proliferative and
carcinogenic potency of estrogens. Since soy products including soy based infant formulas are increasingly being
marketed to children, and to adults for potential health benefits and hormone replacement therapy, we investigated
the effects of early exposure to genistein on the mammary gland. Neonatal CD-1 mice were treated on days 1-5 with
genistein (0.5-50 mg/kg); these doses span the range for human exposures. At 5 and 6 weeks, mammary glands (#4)
were removed and examined by whole mount analysis; estrogen receptor (ER), ER and progesterone receptor
(PR) were studied by immunohistochemistry and/or Western blots. Control mice exhibited an increase in terminal
end buds (TEBs) from 5 to 6 weeks of age indicating active ductal morphogenesis. Mice treated with the lowest
dose of genistein (0.5 mg/kg/day) showed similar increase in the number of TEBs from 5 to 6 weeks but the
numbers were higher than controls at both ages. The genistein 5mg/kg/day dose group showed a different pattern of
mammary gland development with more TEBs present at 5 weeks but less at 6 weeks of age suggesting an advance
in differentiation at this dose. The highest dose of genistein (50 mg/kg/day) showed similar numbers of TEBs as
controls, but mammary gland development was inhibited with smaller ducts and less ductal branching. As another
indication of altered development, the length of the furthest elongated duct was measured; the expanding mammary
ductal mass increased after exposure to genistein 0.5 mg/kg at 5 weeks, and the 0.5 and 5 doses at 6 weeks. In
contrast, mammary glands from the 50 mg/kg/day dose showed growth inhibition of the genistein exposed gland;
this stunted development persisted throughout life.

BASIC POSTER: Female Reproduction II (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                           165
                                                                    Female Reproduction II

P3-15 News Summary
Soy infant formula may negatively affect mammary gland growth, based on animal study

A naturally occurring chemical in soy products – found also in soy infant formula – stunts the
growth of mammary glands in mice, according to a new study being presented on Saturday, June
21, at The Endocrine Society’s 85th Annual Meeting in Philadelphia. The amount of this
chemical, called genistein, to which the mice were exposed is comparable to the amounts of
genistein consumed by young babies drinking soy infant formula.

Although soy has been widely touted for its health benefits, little attention has been paid to any
potential health risks that may result from soy, particularly if exposure occurs during
development. Soy products contain genistein, which is well-known to have estrogenic activity.
Estrogen exposure during development is associated with long-term adverse effects, including a
low incidence of reproductive neoplasia – or tumor formation (exposure to diethylstilbestrol,
DES, is one example).

Therefore, researchers at the National Institutes of Health’s National Institute of Environmental
Health Sciences and the Department of Environmental and Molecular Toxicology at North
Carolina State University, led by Dr. Elizabeth Padilla Banks, investigated the effects of
genistein on developing breast tissue using outbred CD-1 neonatal mice. Genistein treatment
caused a stunting of mammary gland development in mice at levels that are comparable to the
amounts of genistein to which young babies are exposed if they receive soy infant formula.

This adverse finding is one of a series of previously reported abnormalities that Dr. Padilla
Banks and her colleagues have found in mice following developmental exposure to genistein.
Changes in reproductive tract tissues and the ovary have also been described. These data, say the
researchers, add support to the growing concern of exposing young children to soy-based
products – some of which are specifically marketed for children – and infant-based soy formula,
in the absence of medical necessity.

This research was supported by the federal government.

                                              # # #




                                               166
                                                     Mechanisms of Trophoblast Differentiation
OR49-2
A Marked Decline in Coactivators Resulting in a Decrease in Histone Acetylation in the Pregnant Uterus at
Term May Contribute to Inactivation of Progesterone Receptor (PR) Function Leading to the Initiation of
Labor.
Jennifer C Condon*1, Carole R Mendelson1. 1Depts of Biochem and Ob-Gyn, Univ of Texas SW Med Ctr, Dallas,
TX.

Progesterone plays an important role in the maintenance of uterine myometrial quiescence throughout >95% of
pregnancy. In many species, a decline in circulating progesterone occurs prior to the initiation of parturition.
However, in humans, levels of circulating progesterone and of myometrial progesterone receptor (PR) remain
elevated through term. Nevertheless, the finding that PR antagonists can increase uterine contractility in pregnant
women suggests that functional PR inactivation may be involved in the onset of labor. We propose that labor in
women and other mammalian species is initiated by a series of biochemical events that negatively impact PR
function within the myometrium. In the present study, we have tested the hypothesis that a decline in PR
coactivators, leading to a decrease in histone acetylation in the uterus toward term, may contribute to the decline in
PR function, thereby leading to the onset of labor. Myometrial tissues were obtained by cesarean section from the
uterine fundus of women before and after initiation of labor. Uterine tissues were also obtained from pregnant mice
on days 15-19 (term) of gestation. We observed markedly reduced expression of the coactivators CREB binding
protein (CBP) and the steroid receptor co-activators (SRC), SRC-2 and SRC-3, in myometrial tissues of women in
labor as compared to those not in labor. We also found dramatic decreases in expression levels of CBP, SRC-1 and
SRC-3 in mouse uterus at term. The coactivators CBP, SRC-1 and SRC-3 possess histone acetylase activity. To
examine the functional consequence of this decline in coactivators, we analyzed levels of acetylated and total
histone H3 by immunoblotting. Intriguingly, we found a striking decrease in levels of acetylated histone H3 in
fundal myometrium from women in labor as compared to those not in labor. We also observed a dramatic decrease
in levels of acetylated histone H3 in the pregnant mouse uterus at term. Total histone H3 levels were maintained
constant in all samples tested. We suggest that maintenance of elevated levels of histone acetylation within the
myometrium during most of pregnancy promotes an open chromatin structure which facilitates PR activation of
genes that maintain uterine quiescence. At term, the decline in global histone acetylation results in a ‘closing’ of
chromatin structure, resulting in compromised PR function and increased sensitivity of the uterus to contractile
stimuli leading to labor.
          Supported by NIH-3-P01-HD11149-25S1

BASIC ORAL: Mechanisms of Trophoblast Differentiation (2:35 PM - 3:35 PM)

Presentation Date: 6/22/2003, Time: 2:50 PM; Location: 203 A




                                                         167
                                            Mechanisms of Trophoblast Differentiation
OR49-2 News Summary
First-time finding of mechanics of labor may lead to prevention of premature births

Researchers uncover novel mechanism by which progesterone-related proteins help maintain
pregnancy and induce delivery at term, according to a new study being presented on Sunday,
June 22, at The Endocrine Society’s 85th Annual Meeting in Philadelphia. This discovery may
lead to the development of therapeutic strategies to prevent preterm labor and its consequences.
More than 450,000 babies are born prematurely in the United States each year. Significant
numbers of these infants die at birth or suffer lifelong impairment caused by cerebral
hemorrhage. Others develop respiratory distress syndrome because of their poorly developed
lungs, predisposing them to a life of chronic lung disease and potentially other problems.

Despite decades of research, the biochemical mechanisms that cause a woman to go into labor
both prematurely and at term – 40 weeks – remain poorly understood. It is known that
progesterone, a steroid hormone that circulates in the blood at very high levels in the pregnant
woman, plays a critical role in preventing the uterine muscles from contracting until the very end
of pregnancy. In most mammals, progesterone levels, which are high during pregnancy, fall
dramatically just before the onset of labor. This decrease in progesterone is thought to trigger
contractions of the uterus that lead to delivery of the baby. However, in women, the levels of
progesterone do not fall before the onset of labor – creating a scientific mystery for researchers.

Based on these observations, Drs. Jennifer C. Condon and Carole R. Mendelson at the University
of Texas Southwestern Medical Center in Dallas hypothesized that accessory proteins – the so-
called coactivators that help progesterone keep the uterus relaxed – might be reduced at the end
of pregnancy and lead to the induction of labor.

Therefore, they analyzed levels of coactivators in uterine muscle tissues from two groups of
pregnant women at term who were undergoing cesarean section. One group, composed of 16
women, was in labor and the other group of 16 was not. They made a novel observation that
levels of these coactivator proteins were dramatically reduced in the uterine muscle of women
who were in labor, as compared to those who were not in labor. A similar decrease in the
coactivator proteins was observed in the uteri of pregnant mice just before labor. To prove the
link between coactivator decline and labor, they treated pregnant mice with a drug that blocked
the actions of the coactivators. They found that the treated mice failed to go into labor at the
normal time, delivering 24–48 hours late. Based on these findings, the researchers believe that
they have uncovered an important mechanism leading to labor in women and in other mammals.
They suggest that the decrease in coactivator levels in the pregnant uterus at the end of
pregnancy prevents the normal function of progesterone to maintain the uterine muscles in a
relaxed state, leading to increased uterine contractions and delivery of the baby.

This research was supported by the National Institutes of Health and the Texas Higher Education
Coordinating Board.
                                             # # #



                                               168
                                                                                         Male Reproduction
P2-161
Suppression of Spermatogenesis with Levonorgestrel and Testosterone Implants in Asian and Non-Asian
Men.
Ka Kui Lee*1, Ronald S Swerdloff1, Anita Nelson1, Xinghai Wang2, Jian-Sun Tong2, Yu Gui Cui3, Rajiv Roy1,
Robert Sinow1, Nancy Berman1, Laura Hull1, Christina Wang1. 1Depts of Med, OB/GYN, Peds & Radiology,
Harbor-UCLA Med Ctr, Torrance, CA; 2Jiang-su Family Planning Res Inst, Jiangsu, Nanjing, China; 3Dept of Med,
Nanjing Med Univ, Jiangsu, Nanjing, China.

Previous studies have shown that addition of a progestin to an androgen will enhance the rate of suppression of
spermatogenesis in men, To examine whether combination of a long-acting progestin to a long acting androgen
preparation would enhance the efficacy of suppression of spermatogenesis by the androgen alone, 40 non-Asian (in
Los Angeles, USA) and 40 Chinese (in Nanjing, China) healthy male volunteers were randomised to receive either
levonorgestrel (LNG) capsules (2 systems of Norplant II /Jadelle inserted on day 1 delivering approximately 160 ug
LNG per day) together with testosterone (T) implants (4 x 200 mg T pellets inserted on day 1 and week 15
delivering about 6 mg T/day) versus T pellets alone for 30 weeks. The primary endpoint was the % of subjects that
suppressed to very severe oligozoospermia defined as less than 1 million/mL. Prior studies showed that this very
low sperm concentration would result in high contraceptive efficacy. As shown in the figure, the combination of
LNG + T pellets caused firstly a faster suppression of spermatogenesis to a sperm concentration of less than 1
million/mL than T pellets alone in both Asian and non-Asian men.




Secondly, the combination also prevented the drop in spermatogenic suppression observed at week 15 in the T
implant group before the insertion of the next set of pellets suggesting that this dip might be related to insufficient
suppression provided by the T pellets alone. In Asian men severe oligozoospermia was achieved in over 90 % of
men in both treatment groups at week 30. In the non-Asian men severe oligozoospermia was achieved in 56% of the
T alone group, addition of LNG increased this rate to 89%. T pellet extrusion occurred in 5 Asian men. No clinically
significant changes in body weight, lipid profile, hematocrit or PSA were observed throughout the study period.
Addition of a progestin to an androgen alone regimen appeared to induce a faster rate of suppression of
spermatogenesis in both Asian and non-Asian men and enhanced the efficacy of suppression in non-Asian men.

CLINICAL POSTER: Male Reproduction (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                         169
                                                                          Male Reproduction
P2-161 News Summary
Combination of male and female hormones are viable male contraceptive

The combination of progestin and androgen implants would be a safe, effective, low cost,
reversible and a long-acting method of male contraception, according to a new study being
presented on Friday, June 20, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.
The researchers say that the results of the study are important because it is vital that a safe,
effective, relatively low cost, reversible, and long-acting method of contraception for men be made
available worldwide.

Dr. Christina Wang and colleagues at Harbor-UCLA Medical Center in Torrance, Calif., and from
two institutions in Nanjing, China, recruited 80 healthy male volunteers – 40 non-Asian (in Los
Angeles) and 40 Chinese (in Nanjing, China). They randomly received the combination of
progestin and androgen – in pellet form inserted under the skin – or androgen alone – also in pellet
form inserted under the skin – for 30 weeks.

Results showed that the combination of progestin and androgen suppressed sperm production
faster in both groups than use of androgen alone. It also showed that the combination enhanced
the efficacy of suppression in non-Asian men. No clinically significant side effects were noted,
although the pellet extruded from five Asian men.

The researchers note that the creation of an effective and culturally accepted male contraceptive
would have a profound effect on family planning, giving couples a choice of which partner should
be responsible for birth control. In cases where a woman is advised not to take birth control pills
or finds other methods unacceptable, the man could take responsibility.

The investigators of the study are pioneers in the development of male contraception based on
hormonal methods and have performed many previous studies of male contraceptive
development.

The study supported by the Contraceptive Research and Development Program and the Mellon
Foundation.

                                              # # #




                                               170
                                                                                      Male Reproduction
P2-166
Does Pretreatment Serum Testosterone Affect Responsiveness to Long Term Transdermal Testosterone Gel
(Androgel) Treatment in “Hypogonadal” Men?
Christina Wang*1, Ronald S Swerdloff1, Nancy Berman1, Ali Iranmanesh2, Adrian S Dobs3, Peter J Snyder4, Glenn
Cunningham5, Alvin M Matsumoto6, Thomas Weber7, Testosterone Gel Study Group8. 1Depts of Med & Peds,
Harbor-UCLA Med Ctr & Res & Educ Inst, Torrance, CA; 2Dept of Med, VA Med Ctr, Salem, VA; 3Dept of Med,
Johns Hopkins Univ, Baltimore, MD; 4Dept of Med, Univ of Pennsylvania Sch of Med, Philadelphia, PA; 5Dept of
Med, Baylor Univ Sch of Med, Houston, TX; 6Dept of Med, Univ of Washington, Seattle, WA; 7Dept of Med, Duke
Univ Med Ctr, Durham, NC; 8Dept of Med, Unimed Pharmaceuticals Inc, Deerfield, IL.

We have previously shown that transdermal testosterone gel (Androgel) improves sexual function, mood, body
composition and bone mineral density (BMD) in hypogonadal men. In these studies, the entry criteria included a
serum testosterone (T) <300 ng/dL while off T replacement for 6 weeks measured in the laboratory of each center.
However on the day before Androgel application, about 30% of the men had baseline serum T levels above this
threshold when measured by a central laboratory. We compared the effects of treatment with T gel for up to 36
months in 76 hypogonadal men (mean serum T 154 ± 11 ng/dL, low T group) and another 48 men who were
diagnosed to have hypogonadism yet had baseline serum T levels of >300 ng/dL (398 ± 10 ng/dL, normal T group).
These men with the higher baseline serum T levels also had significantly higher baseline serum DHT, E2 and free T
levels. Initially the subjects were treated with Androderm patch 5 mg/day, Androgel 5 or 10 g /day, dose adjustment
to 7.5 g /day occurred in some men at 3 months to maintain serum T within the eugonadal range. After 6 months
treatment, all subjects were switched to Androgel treatment and dose adjustments were allowed throughout
treatment depending on the symptoms of the subjects while maintaining serum T concentration within the eugonadal
range. Mean serum T, DHT, E2 and free T concentrations were lower (by definition) in the hypogonadal men at
baseline (p < 0.03), they were similar and in the eugonadal range in the two groups after Androgel treatment. There
were no significant differences in age, final dose of Androgel and ethnic distribution between the 2 groups. Sexual
desire was not different between the groups at baseline and throughout treatment. Lean mass was significantly lower
(low T:58.3 ± 1.0, normal T: 62.0 ± 1.3 Kg, p<0.03) and the fat mass (low T:27.5 ± 1.2, normal T: 24.5 ± 1.0 Kg,
p<0.01) higher in the hypogonadal men. With Androgel treatment, the changes in lean (low T: 2.7 ± 0.5, normal T:
2.5 ± 0.6 Kg) and fat (low T: -1.6 ± 0.7, normal T: -1.4 ± 0.6 Kg)mass were not significantly different between the
two groups, BMD of the hip and spine at baseline and after treatment were also not different between the two
groups. We conclude from these data that the baseline serum T concentrations did not significantly affect the
responsiveness of sexual function, body composition and BMD to long term Androgel treatment.
          Supported by grants from UNIMED/Solvay Pharmaceuticals

CLINICAL POSTER: Male Reproduction (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                       171
                                                                             Male Reproduction
P2-166 News Summary
Men with hormone disorder respond well to testosterone gel, regardless of testosterone
levels at treatment

Men with sex hormone disorder with varying levels of testosterone all respond equally well to
treatment with testosterone topical gel, according to a new study being presented on Friday, June
20, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

A normal level of testosterone is important to the health of all men. Testosterone deficiency can
reduce feelings of sexual desire and the ability to maintain an erection, decrease bone density and
cause changes in body composition, including a reduction of muscle mass and increased body
fat. Testosterone replacement therapy is used to improve energy, stabilize or increase bone
density, improve mood, improve muscle strength and body composition and increase libido and
sexual function in men with hypogonadism, a reduced or absent secretion of hormones from the
sex glands.

In this multi-center study, researchers, led by Dr. Christina Wang, compared the effects of
treatment with testosterone gel for up to 36 months in 76 hypogonadal men whose testosterone
levels were repeatedly below those of healthy men. They also treated another 48 men who were
diagnosed to have hypogonadism based on initially low blood testosterone levels but, on repeat,
the majority had levels in the low normal range. All were treated similarly with a transdermal
testosterone gel (AndroGel) after the initial six months. Doses were adjusted throughout
treatment to maintain testosterone levels in the mid-normal range.

The researchers suspected that the men with persistently low blood testosterone levels would
respond better to a rise in testosterone levels than those with variably low baseline levels.
However, this proved not to be the case. The results indicated that both sub-groups improved
and that, in men selected for an initial low testosterone level, the repeat testosterone levels did
not significantly predict the positive responsiveness of sexual function, body composition and
bone mineral density in men to long-term testosterone treatment via a testosterone gel
(AndroGel) applied on the skin. Results showed that there were no significant differences
between the two groups at the end of the study.

This study was performed at 15 centers in the United States. The Male Reproductive Research
Center at the Research and Education Institute at Harbor-UCLA Medical Center is the lead
center for this study.

The study was supported by Unimed-Solvay.

                                                # # #




                                                 172
                                                                                       Male Reproduction
P2-162
Sustained Gonadotropin Suppression with Subcutaneous Testosterone Microcapsule Injections in Normal
Men: A Potential Agent for Male Hormonal Contraception.
Andrea D Coviello*1, Janet K Gilchriest2, Brad D Anawalt2, Alvin M Matsumoto2. 1Med, Univ of Washington,
Seattle, WA; 2Med, VA Puget Sound Hlth Care Syst, Seattle, WA.

Background: In male hormonal contraceptive regimens exogenous testosterone (T) suppresses spermatogenesis by
inhibition of gonadotropins. Currently available T esters require intramuscular injection. A sustained release
preparation consisting of T complexed to a biodegradable polylactide-co-glycolide polymer capsule (T
microcapsule) delivered by subcutaneous (sc) injection may provide a more practical and acceptable method of T
administration as well as sustained gonadotropin inhibition. We hypothesized that a single sc injection of T
microcapsules would raise serum T resulting in prolonged suppression of LH and FSH. Methods: 6 healthy men
(age 20-44) were given 534 mg of T microcapsules (supplied by Biotek, Woburn, MA) by sc injections in the
buttocks. Serum T, LH, and FSH were determined on days 0, 1, 2, 4, 7, 14, 28, 56, and 84 after T injection.
Hematocrit, fasting lipids, and prostate specific antigen (PSA) were determined on days 0, 28, 56, and 84. Sperm
counts were determined on days 0, 56, and 84. Mean changes from baseline over time were assessed by paired t-test.
Results: Mean T increased 244% (19.4 to 47.3 nmol/L) in the first 24 hours post injection and returned to baseline
by day 28 (17.6 nmol/L). LH was suppressed to < 1 mIU/ml by day 4 (p < 0.01) and FSH was suppressed by day 7
(p < 0.01). Both remained significantly suppressed through day 28 (p < 0.01). Mean sperm count decreased from 69
million/ml at baseline to 39 million/ml at day 56 and to 13 million/ml at day 84 (p = 0.01) despite the return of
serum T and LH/FSH to baseline by day 84. There was no significant change in weight, hematocrit, lipids, or PSA.
Men experienced moderate pain at the site of injection which resolved spontaneously within a week. Summary: 1)
T delivery by sustained release microcapsules results in an increase in serum T that induces gonadotropin
suppression within one week which is maintained for one month in normal men. 2) Sperm production was
suppressed at 2 and 3 months after a single injection. 3) Although men experienced transient discomfort at injection
sites, there were no significant side effects. Conclusion: Microcapsule formulations of T plus progestins and/or
gonadotropin releasing hormone antagonists may be a practical and safe male hormonal contraceptive.

CLINICAL POSTER: Male Reproduction (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                       173
                                                                           Male Reproduction
P2-162 News Summary
Sustained-released, testosterone male contraceptive appears safe, effective for future use
A new male contraceptive that releases testosterone over three months is potentially a safe and
practical method of contraception, according to a new study being presented on Friday, June 20,
at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

There have been ongoing efforts to find safe, effective, hormone-reversible contraceptive options
for men to expand their options outside of condoms or vasectomy. Testosterone has previously
been shown to effectively suppress sperm production and is an essential component of male
hormonal contraceptives to both suppress sperm production and to replace testosterone normally
produced by the testes. However, the vast majority of testosterone preparations that have been
used in contraceptive research protocols are injected intramuscularly on a weekly basis. There
are ongoing efforts by researchers to develop less painful and more practical methods of
delivering testosterone. A new sustained-release testosterone microcapsule – a thick liquid
administered by injection under the skin – has been developed that alters testosterone to a
biodegradable polymer, releasing testosterone over three months.

To evaluate testosterone microcapsules for potential use in a male contraceptive, Dr. Andrea
Coviello and colleagues in Seattle gave subcutaneous injections to six healthy normal men and
followed their sperm counts for three months. Testosterone microcapsules successfully
suppressed the hormones responsible for stimulating sperm production. Sperm counts
significantly decreased (81 percent) from measurements at the beginning of the study. Even
though there was some pain at the injection site, there were no serious side effects. The
prolonged suppression of sperm production after a single injection suggests that testosterone
microcapsules may be a safe, practical method of testosterone delivery in male contraceptive
regimens.

In addition, Dr. Coviello and colleagues at the University of Washington are participating in a
study of thin, contraceptive, rod-like inserts that are implanted under the skin in men and are also
planning to test other non-injection formulations in the near future.

This study was supported by the Contraceptive Research and Development Program in
Arlington, Va., and Biotek, Inc. in Woburn, Mass., who also supplied the testosterone
microcapsule drug.

                                              # # #




                                                174
                                                                  Reproductive Biology in the Male
P1-202
Effects of Androgens on Coronary Heart Risk Profile and the Relation to Male Pattern Baldness: A Study in
Female-to-Male Transsexuals.
Arno WFT Toorians*1, Erik J Giltay2, Angela R Sarabdjitsingh1, Nienke A de Vries1, Louis JG Gooren1.
1
  Endocrinology/Androl, Vrije Univ Univ Med Ctr, Amsterdam, Netherlands; 2Div of Hum Nutrit & Epidemiology,
Wageningen Univ, Wageningen, Netherlands.

There is a sex difference in the incidence of coronary heart disease (CHD) traditionally attributed to the higher
androgen levels in men compared to women and their effects on lipid profiles. Male-pattern baldness (MPB) has
been associated with CHD; MPB is one of the virilizing effects of androgens, and might serve as a marker of
androgen-induced effects on risk factors of CHD, such as increases in LDL cholesterol and reductions in HDL
cholesterol, and other markers of CHD.
We studied 86 eugonadal female-to-male transsexuals (FMTr), aged 27 years (range 16-48), treated with
intramuscular testosterone esters (Sustanon®, n=64, 250 mg/2 weeks) or oral testosterone undecoanoate (Andriol®,
n=22, 160-240 mg/day). At baseline and during follow up sex hormones, body mass index, blood pressure, smoking
status were assessed, as well as lipid, glucose, and insulin levels. The risk of CHD was estimated by the
Framingham and PROCAM models. Baldness was self-assessed using a modified 5-point Hamilton Scale.
Androgen induced baldness (score II to V) was observed in 34 of 86 participants (39.5%). Initial laboratory values
of sex steroids and cardiovascular risk markers and the shifts induced by androgen administration were not different
between ‘non-bald’ vs. ‘bald’ participants. Coronary risk increased from median 0.94 per 100,000 females per 8
years (p25 0.38 to p75 3.55) to 1.79 (0.92 – 7.29; P<0.001) according the PROCAM model, and from 0.03 (0.00 –
1.37) to 0.87 (0.10 - 11.17) according the Framingham model (P<0.001 for both).The estimated relative risk of CHD
clearly increases following androgen administration to young FM transsexuals, yet the absolute estimated risk
remains low. MPB induced by androgen administration was not associated with stronger shifts in endocrine and
CHD risk variables. This finding does not support a hypothesis of shared androgen pathways of androgenetic
alopecia and CHD risk factors.

BASIC POSTER: Reproductive Biology in the Male (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                       175
                                                        Reproductive Biology in the Male

P1-202 News Summary
Contrary to previous research findings, male baldness is not a sign of heart disease risk

Despite an association between male hormones, balding and heart disease risk factors,
researchers found that male pattern baldness (MPB) is not an indicator of an increased risk of
heart disease, according to a new study being presented on Thursday, June 19, at The Endocrine
Society’s 85th Annual Meeting in Philadelphia.

The scalp sensitivity to androgens – male sex hormones, such as testosterone – is part of the
pathophysiology of MPB. Established risk factors for heart disease are also affected by
androgens. Therefore, researchers from the Netherlands hypothesized that the association
between androgens and heart disease risk factors might explain the epidemiology association
between MPB and heart disease.

In a retrospective, observational design, Dr. Arno Toorians and his colleagues studied 81 female-
to-male transsexual subjects, with an average age of 27 (range 16–48), treated with two forms of
testosterone: intramuscluar esters (na; 250mg IM/2wks) or oral undecanoate (n; 160–240 mg/d
orally). The degree of MPB was self assessed using a five-point scale. For example, type I on
the scale was “no hair loss” and type V was “complete hair loss.” Body mass index, blood
pressure and levels of lipid and insulin were assessed at the start of testosterone administration,
between six to 24 years before, and at three-to-four months after the start of the treatment with
testosterone.

Of the 81 transsexuals, 31 (38.3 percent) had MPB from type II to type V. Thinning of hair was
related to the duration of androgen administration and present in 50 percent of androgen-treated
female-to-male transsexuals after approximately 13 years. None of the heart disease risk factors
or proportional changes were associated with the degree of MPB at follow up, except that there
was an unexpected tendency of lower fasting glucose in balding subjects.

The researchers findings, therefore, do not support the idea that MPB serves as an indicator of
increased heart disease risk through androgenic effects on classical heart disease risk factors.

                                              # # #




                                               176
                                                                   Reproductive Biology in the Male
P1-183
Erectile Dysfunction in Men with Diabetes Mellitus: How Safe Is Sildenafil Citrate?
Cornelia Jaursch-Hancke*1, Walter Merkle1, Hans-Peter Tries1, Heinrich Lambertz1, Dietz Ulrich1. 1German Diag
Clin, Wiesbaden, Germany.

Introduction:
Erectile Dysfunction (ED) is common in men with diabetes mellitus. It is suggested that oral Sildenafil citrate is
well-tolerated and effective in improving ED in diabetic men (1). Before treatment with Sildenafil a treadmill ECG
at a 100W level is recommended to assess cardiovascular health (2).
The aim of the study was to evaluate the cardiovascular health in men with diabetes and investigate the effects of
Sildenafil on coronary flow dynamics (CFR) of the left anterior descending artery.
Patients and Methods:
Fourty three otherwise healthy diabetic men (aged 59 +/-7 years) with ED and without symptoms of coronary artery
disease were included. In each patient a treadmill ECG and echocardiography were performed. In case of
unremarkable cardiological evaluation CFR was done.
Results:
ECG- treadmill at a 100 W level were normal in all men (n=43). Because of clinical suspicion (n=11), CFR < 200%
(n=2) and pathologic ECG-treadmill at a higher W level (n=3) coronary angiography was performed. Severe
coronary artery disease was confirmed in 12 patients who wee excluded from further analyses. In 10 patients CFR
was not possible because of technical reasons. CFR at baseline was at the lower of the normal range in 17/21
diabetics (median 245%, range 210-490%). CFR decreased insignificantly in 12/21 patients after Sildenafil
administration ( delta CFR -10%, p=0,3).
Conclusion:
Asymptomatic severe coronary artery disease was found in about 25% of diabetic men. Sexual activity after
treatment with Sildenafil could be a risk factor for cardiovascular events in this group. Diabetics with ED often have
a CFR in the lower normal range, but Sildenafil did not further reduce CFR.
         Reference: (1) Rendell MS et al. Sildenafil for threatment of erectile dysfunction in men with diabetes,
JAMA 1999, 281 (5): 421-466
         (2) Cheitlin MD et al. Use of Sildenafil (Viagra®) in patients with cardiovascular disease, Circulation
1999, 99: 168-177

BASIC POSTER: Reproductive Biology in the Male (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                         177
                                                        Reproductive Biology in the Male
P1-183 News Summary
Sexual activity after taking sildenafil (Viagra®) may not be heart safe in some diabetic men

Researchers found that sildenafil did not significantly affect heart function in diabetic men;
however, cardiac evaluation revealed severe heart disease without symptoms in one-quarter of
this group, according to a new study being presented on Thursday, June 19, at The Endocrine
Society’s 85th Annual Meeting in Philadelphia. This underlying but silent disease could be a
harmful combination with sildenafil, say the researchers.

Sexual dysfunction is common in men with diabetes. Oral agents like sildenafil are comfortable
and effective in improving erectile dysfunction (ED) in diabetic men; however, there have been
many studies warning of potential problems in the case of certain heart conditions.

Therefore, Dr. Cornelia Jaursch-Hancke and colleagues at the German Diagnostic Clinic in
Wiesbaden, Germany, investigated cardiovascular health in 43 diabetic but otherwise healthy
men (52–66 years old) before and after treatment with sildenafil. In each patient, a treadmill
electrocardiogram and echocardiography were performed.

The researchers found that sildenafil did not significantly influence heart function. However,
careful cardiac evaluation revealed asymptomatic, severe cardiac disease in 25 percent of the
diabetic men. Therefore, the researchers concluded that sexual activity after treatment with
sildenafil could be a risk factor for heart attacks in diabetic men.

                                             # # #




                                               178
                                                                   Reproductive Biology in the Male
P1-186
The Phytoestrogen Metabolite Equol Acts as a Novel Anti-Androgen To Inhibit Prostate Growth and
Hormone Feedback.
Trent D Lund*1, Daniel J Munson1, Megan E Haldy1, Edwin D Lephart2, Robert J Handa1. 1Biomed Scis, Colorado
State Univ, Fort Collins, CO; 2The Neurosci Ctr and Dept of Physiol and Devel Biol, Brigham Young Univ, Provo,
UT.

Equol represents the major metabolite of the phytoestrogen daidzein, an isoflavone found abundantly in soy.
Because of its protective effects in cardiovascular, bone and menopausal health equol is heralded as an important
isoflavone in humans, although it is produced by the gut bacteria of only some individuals. As shown by the studies
presented here, one aspect of equol’s mechanism of action is apparently its ability to prevent 5-alpha
dihydrotestosterone’s (DHT) actions in physiological processes. To examine the effects of equol on androgenic
processes we administered equol (0.25-0.5 mg/kg) to intact adult male rats for 4-7 days. This treatment caused a
reduction in prostate and epididymal weight without changing testicular, pituitary, or body weights. Another group
of male rats were castrated (GDX) and then treated, 7 days later, with DHT or DHT+equol. Equol blocked DHT’s
trophic effects on the prostate gland. Furthermore when administered to intact animals, equol significantly increased
LH levels vs. DMSO treated control values indicating a blockade of negative feedback. Similarly, when GDX males
were treated with equol +/-DHT it completely reversed the inhibitory action of DHT on plasma LH levels. DHT
treated males had low levels of LH, whereas DHT+equol-treated male rats displayed high LH levels similar to that
of GDX controls. These data suggest that equol has the ability to block the negative feedback action of DHT. In
these studies equol’s inhibitory effects occurred without reducing plasma DHT levels. Males injected with DHT in
combination with equol actually had plasma DHT levels that were elevated above those animals treated with DHT
alone. Equol’s ability to inhibit DHT action has important ramifications in health and disease and may indicate a
broad and important usage for equol in the treatment of androgen mediated pathologies. The possible mechanisms
for equol’s action are presented in a related abstract (Lund et al, The Phytoestrogen Metabolite Equol Acts as a
Novel Anti-Androgen by Binding DHT, Endo Abstracts, 2003). Given the high consumption of phytoestrogens in
Asian populations, our findings correlate with the low incidence rates of benign prostate hyperplasia/prostate cancer
in Asian men compared to Western populations and suggest a mechanism for such epidemiology. These data also
provide a potential avenue for the therapeutic use of equol in the treatment of androgen-mediated pathologies.
         Supported by NIH grants AA12693 and NS39951 (RJH).

BASIC POSTER: Reproductive Biology in the Male (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                        179
                                                        Reproductive Biology in the Male
P1-186 News Summary
By-product of naturally occurring estrogen may help male hormone–related diseases

A by-product of a naturally occurring estrogen, called equol, can block negative actions of
metabolized testosterone, which can promote a number of health problems, according to a study
being presented on Thursday, June 19, at The Endocrine Society’s 85th Annual Meeting in
Philadelphia. These data may lead to the therapeutic use of equol in treating male hormone–
mediated diseases, such as prostate cancer and benign prostate hyperplasia (BPH).

Phytoestrogens are naturally occurring estrogen-like molecules found abundantly in soy. One
type of phytoestrogen, daidzein, can be converted by intestinal bacteria, found in about 30
percent of the human population, to another potent compound – equol. Studies showing
protective effects of equol in cardiovascular, bone and menopausal health indicate that it may be
an important consideration in human health and disease. In contrast to all previous studies
examining equol’s estrogenic properties, Dr. Trent D. Lund and colleagues in Fort Collins,
Colo., and Provo, Utah, uncovered a unique aspect of equol’s mechanism of action.

In this study, Dr. Lund and colleagues examined the effects of equol administration on certain
androgen-mediated functions. First, equol was administered to intact adult male rats for four to
seven days. This treatment caused a reduction in prostate gland and epididymal weights without
changing testicular, pituitary or body weights. Both prostate and epididymal growth have been
linked to their ability to metabolize testosterone to dihydrotestosterone (DHT).

DHT is a potent and extremely active metabolite of the male hormone, testosterone. It has been
linked to a number of health problems, including benign prostate hyperplasia and prostate
cancer; female- and male-pattern baldness; facial and body hair growth; acne, aging and photo
aging; skin integrity like collagen and elastin robustness and emotional and mental health issues.

Subsequently, another group of male rats were castrated and then treated with DHT or DHT and
equol. Equol treatment blocked DHT’s growth promoting effects on the prostate gland.
Furthermore, equol treatment of intact male rats significantly increased luteinizing hormone
(LH) levels compared to the control group. This finding indicates that equol can block the
negative feedback actions of androgens on pituitary hormone secretion. Similarly, when
castrated male rats were treated with equol at the same time as DHT, it completely prevented the
inhibitory action of DHT on plasma LH secretion. Equol’s anti-androgen actions occurred
without reducing circulating plasma DHT levels. In fact, male rats injected with DHT and equol
had higher plasma DHT levels than those animals treated with DHT alone.

Given the high consumption of phytoestrogens in Asian populations, researchers say that these
findings correlate with the low incidence of PBH and prostate cancer in Asian versus Western
populations and suggest a mechanism for such epidemiology.

This research was supported by the National Institutes of Health.
                                             # # #


                                               180
                                    Inhibin, Activin & Transforming Growth Factor-beta
P2-153
Reduction in Cardiac Muscle Mass in Transgenic Mice That Hyperexpress Myostatin Suggests a Role of
Myostatin in Cardiomyogenesis.
Jorge N Artaza*1, Shalender Bhasin1, Suzanne Reisz-Porszasz1, Nestor F Gonzalez Cadavid1. 1Div of
Endocrinology and RCMI DNA Repository and Molec Med Core., Charles Drew Univ, Los Angeles, CA.

Inactivating mutations of myostatin (Mst) are associated with hypermuscularity, indicating that Mst is a negative
regulator of skeletal muscle growth; we do not know whether Mst also regulates cardiac muscle mass. To elucidate
the role of Mst in cardiomyogenesis, we studied the effects of Mst hyperexpression on cardiac muscle mass in
transgenic mice (TG) in which wild type muscle creatine kinase (MCK) promoter was used to drive hyperexpression
of Mst in skeletal and cardiac muscle. TG harboring this MCK promoter upstream of EGFP protein cDNA
expressed fluorescence only in skeletal and cardiac muscles, demonstrating the ability of MCK promoter to restrict
transgene expression to these tissues. Mst mRNA and protein expression, measured by RT-PCR and western blot,
were higher in cardiac muscle of male and female TG, in comparison to wild type mice (WT). Male Mst-TG mice
had, in comparison to WT (n=11), significantly lower total cardiac muscle mass (110.9±5.7 vs 134.3±7.1 mg,
p<0.01); left ventricular mass (LV) corrected by body weight (1.77±0.06 vs 2.08±0.12 mg/g, p<0.01); and left atrial
mass (2.26+/-0.20 vs 3.14±0.42 mg, p<0.05). Cardiac muscle mass (95.3±2.9 vs 95.4±4.6 mg), and individual
chamber masses in female Mst-TG mice were not significantly different from WT mice even though Mst mRNA
and protein contents were higher. To determine whether decreased cardiac muscle mass in Mst-TG male mice was
due to increased expression of Mst transgene in cardiac muscle or to the endocrine effects of circulating Mst, we
generated another TG mouse in which Mst trasngene hyperexpression was limited to skeletal muscle, by using a
mutated form of the MCK promoter (MCK-3E-Mst); this promoter directs expression only in the skeletal muscle but
NOT in cardiac muscle. Although skeletal muscle mass was significantly lower in MCK-3E-Mst TG mice compared
to WT mice, the cardiac muscle mass was not significantly different between MCK-3E-Mst and WT mice.
Conclusions: Increased expression of Mst in the heart is associated with lower cardiac muscle mass due to a
reduction in left ventricular and atrial mass, suggesting that myostatin also regulates cardiac muscle growth.
Increased myocardial expression of Mst is necessary for inhibition of cardiac muscle mass because this effect was
not observed in MCK-3E-Mst mice in which Mst hyper-expression was limited to skeletal muscle. The gender
differences and the mechanisms by which Mst regulates cardiomyogenesis should be investigated.

BASIC POSTER: Inhibin, Activin & Transforming Growth Factor-beta (11:00 AM - 12:00 PM and 2:30 PM - 3:30
PM)

Presentation Date: Friday, June 20, 2003




                                                       181
                               Inhibin, Activin & Transforming Growth Factor-beta

P2-153 News Summary
Novel protein shown to affect heart muscle growth; may hold keys to future heart therapy

The protein myostatin, which has been shown to block the development of muscle mass, may
also be an important factor in cardiac muscle growth, according to a study being presented on
Friday, June 20, at The Endocrine Society’s 85th Annual Meeting in Philadelphia. Based on this
early animal research, myostatin may be an important factor preventing heart cells to regrow and
repair damaged tissue after a heart attack.

Cardiac tissue repair after a heart attack is limited by the inability of heart cells – called
cardiomyocytes – to proliferate during regeneration. This tissue can only be repaired by the
growth of existing cells, and factors that impede cardiomyocyte proliferation are unknown.

Myostatin is a protein that is a novel negative regulator of skeletal muscle mass, and laboratory
mice that have been genetically engineered to block the production of myostatin (“knock-out”
mice) have much higher skeletal muscle mass than wild-type animals. The same occurs with
certain breeds of cattle that harbor an inactivating mutation of the myostatin gene. Myostatin is
over expressed in conditions associated with muscle wasting and after experimental damage to
the heart in laboratory animals.

Researchers at Charles Drew University in Los Angeles, led by Dr. Jorge Artaza, constructed a
genetically altered, or transgenic, mouse that, in contrast to the myostatin knock-out mouse, over
produced myostatin in the skeletal muscle and heart. As a result, the skeletal muscle mass was
reduced by nearly 20 percent compared to the wild-type animal, but it occurred only in males.

To determine whether myostatin is one of the factors that blocks the replication of
cardiomyocytes and limits heart tissue regeneration, the researchers measured cardiac muscle
mass in the transgenic mouse and found that there was also a reduction in the size of the heart,
specifically of the left ventricle, and it occurred only in the male. To confirm this finding, the
researchers created another transgenic mouse that over expressed myostatin only in the skeletal
muscle and not in the heart and found that this mouse did not exhibit a smaller heart, despite
reduced skeletal muscle.

However, the researchers note that further work must be done to determine if myostatin affects
cardiac function, whether this growth inhibitor can really stop the replication of cardiomyocytes,
and what the reason is for the apparent gender differences in myostatin action. Once these
questions are answered, future studies can examine if myostatin is a key inhibitor of cardiac
regeneration after a heart attack or if it plays a role in the excessive growth of extraneous
connective tissue in the heart. Should this be the case, they add, then future therapies may try to
block myostatin effects in the heart.

The research was supported by the National Institutes of Health.
                                            # # #


                                                182
Steroid Hormone Biosynthesis/Action
                                                          Steroid Hormone Action & Metabolism
P3-221
Pharmacologic Effects of a Novel Selective Androgen Receptor Modulator (SARM), Flutamide and
Finasteride in Intact Male Rats.
Wenqing Gao*1, Jeffrey D Kearbey1, Kiwon Chung3, Duane D Miller2,3, James T Dalton1,3. 1Div of Pharmaceutics,
Coll of Pharmacy, The Ohio State Univ, Columbus, OH; 2Dept of Pharmaceutical Scis, Coll of Pharmacy, Univ of
Tennessee, Memphis, TN; 3GTx Inc., Memphis, TN.

The purpose of this study was to compare the pharmacologic effects and tissue-selectivity of flutamide, finasteride
and a novel selective androgen receptor modulator in intact male rats. Intact male Sprague-Dawley rats were treated
with flutamide (5 mg/kg), finasteride (5 mg/kg), SARM (5mg/kg) or vehicle for 3, 6, or 9 days. Castrated rats were
also included as controls for each time point. The drugs were administered daily via subcutaneous injections. The
weights of androgenic (i.e., prostates and seminal vesicles) and anabolic (i.e., levator ani muscle) tissues in animals
of different treatment groups were measured at the end of each treatment period, and were normalized with the body
weights. Percentages were determined by comparison to intact animals. In castrated control groups, prostate,
seminal vesicles, and levator ani muscle weights decreased gradually from 45%, 30% and 71%, to 15%, 14% and
62%, respectively, between day 3 and day 9. Flutamide significantly decreased the prostate, seminal vesicles, and
levator ani muscle weights in intact male rats to 65%, 63% and 84%, respectively, with no significant change over
time. However, both finasteride and our SARM showed significant decreases in prostate and seminal vesicle without
affecting the levator ani muscle. Finasteride significantly decreased the prostate and seminal vesicle weights to 55%
and 29% respectively, with no significant change over time. Our SARM gradually decreased the prostate and
seminal vesicle weights from 79% and 74%, to 48% and 42%, respectively. In summary, this SARM can effectively
decrease the weights of the prostate and seminal vesicles without affecting the levator ani muscle in intact male rats.
These data suggest that a non-steroidal SARM may be useful in the treatment of benign prostatic hyperplasia (BPH).
          Supported by a grant from GTx, Inc.

BASIC POSTER: Steroid Hormone Action & Metabolism (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                         183
                                                 Steroid Hormone Action & Metabolism
P3-221 News Summary
New class of non-steroidal drugs may be superior treatment for common prostate problem

Non-steroidal anti-androgens may be useful in the treatment of benign prostatic hyperplasia
(BPH), overcoming limitations of other therapies by both shrinking the prostate and preventing
the progression of BPH, according to a study being presented on Saturday, June 21, at The
Endocrine Society’s 85th Annual Meeting in Philadelphia. This new class of drugs also shows
potential for treating muscle wasting and osteoporosis or as a male contraceptive.

More than 50 percent of men older than 60 years of age have symptoms of BPH – a condition in
which non-cancerous nodules enlarge the prostate gland. Symptoms include urinary hesitancy,
pain with urination, bloody urine, increased urinary frequency and incontinence.

The most popular medical therapies currently include α-adrenergic blockers to relieve these
symptoms. Unfortunately, α-adrenergic blockers do not shrink or prevent the progression of
BPH. Androgen receptor antagonists, like flutamide, are effective in this regard, but they have
unfavorable side effects, including muscle wasting, decreased sexual libido and impotence.
Another class of drugs, 5 α reductase inhibitors, does not block the androgen receptor, thereby
allowing testosterone to bypass this enzyme and stimulate the prostate. Nonsteroidal selective
androgen receptor modulators (SARMs) are tissue selective and antagonize testosterone’s
growth, promoting activities in the prostate, but are able to maintain muscle, bone and libido.

Researchers at Ohio State University in Columbus and the University of Tennessee in Memphis
– led by senior investigator Dr. James T. Dalton – compared the pharmacologic effects and
tissue-selectivity of flutamide, finasteride and a novel SARM in male rats. The rats were divided
into four groups, receiving one of the three drugs or a placebo for three, six or nine days.
Castrated rats were also included as controls for each time point. The drugs were administered
daily via subcutaneous injections. The weights of androgenic (i.e., prostates and seminal
vesicles) and anabolic tissues (i.e., levator ani muscle, which is a broad, thin muscle on the inner
service of the pelvis) in animals of different treatment groups were measured at the end of each
treatment period and were normalized with the body weights.

In the castrated control groups, prostate, seminal vesicles and levator ani muscle weights
decreased gradually from 45 percent, 30 percent and 71 percent to 15 percent, 14 percent and 62
percent, respectively. Flutamide significantly decreased the prostate, seminal vesicles and
levator ani muscle weights in intact male rats to 65 percent, 63 percent and 84 percent,
respectively. However, both finasteride and the SARM showed significant decreases in prostate
and seminal vesicle without affecting the levator ani muscle. Finasteride significantly decreased
the prostate and seminal vesicle weights to 55 percent and 29 percent, respectively. The SARM
gradually decreased the prostate and seminal vesicle weights from 79 percent and 74 percent to
48 percent and 42 percent, respectively.

This research was supported by GTx, Inc.
                                              # # #


                                                184
                                                           Steroid Hormone Action & Metabolism
P3-196
The Influence of Classical Yoga Practices on Plasma Cortisol Levels.
Justin Mager*1, Vijayendra Pratap2, Barbara Levitt2, John Hanifin1, George Brainard1. 1Ctr for Integrative Med,
Jefferson Med Coll, Philadelphia, PA; 2Yoga Res Soc, Philadelphia, PA.

In modern western culture, yoga has become increasingly popular as a vehicle for coping with stress. A preliminary
report (Brainard et al., 77th Meeting of the Endocrine Society, 1995) suggested that classical yoga may have a
specific influence on cortisol regulation in healthy humans. The specific aims of the following study were: 1) to
repeat the experiment to test if a single set of classical yoga practices can lower plasma cortisol levels in
inexperienced practitioners of yoga; and 2) to extend the experiment to test if 7 continuous days of yoga practices
can lower plasma cortisol levels in inexperienced practitioners of yoga. Healthy female and male volunteers (n=16,
mean age 27.6 ± 1.0) were recruited for a within-subjects designed experiment. All subjects were yoga naive,
defined as having one or less formal yoga classes within one year of participation and no past history of a consistent
yoga practice. On day 1 of the eight day study, volunteers sat quietly in chairs, reading or writing from 11:00 AM
until 11:50 AM. Blood samples (20 ml) were collected from all volunteers immediately before and after this control
session. On day 2 and 8 of the study, subjects were guided through a 50 minute set of yoga practices, from 11:00
AM until 11:50 AM, by an experienced classical yoga instructor. Again, blood samples (20 ml) were collected
immediately before and after the yoga practice sessions. During days 3 through 7, subjects were guided through a
50 minute set of yoga practices, each day from 5:30 PM until 6:20 PM. No blood samples were collected during
days 3 through 7. Plasma cortisol levels were quantified by RIA. Analysis of variance showed there were no
significant differences between plasma cortisol levels on days 1, 2 and 8 for the first time point (F= 0.47; P = 0.63).
A decrease in plasma cortisol levels from 11:00 AM until 12:00 PM were observed in 42 of the 48 samples collected
at the second time point, which is consistent with the normal circadian changes in cortisol. Paired t-tests showed
that these decreases were statistically significant on the yoga practice days (t = 4.43, p < 0.001 on day 2; t = 4.34, p
< 0.001 on day 8) but were not significant on the control day (t = 1.65, p = 0.12 on day 1). Data from the current
study demonstrate that classical yoga practices are associated with significantly lower plasma cortisol levels in
healthy humans after one day and one week of practice.

BASIC POSTER: Steroid Hormone Action & Metabolism (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                         185
                                                 Steroid Hormone Action & Metabolism
P3-196 News Summary
Yoga reduces levels of stress-related hormone

Yoga induces significant decreases in the stress-related hormone cortisol in healthy,
inexperienced practitioners of yoga, according to a study being presented on Saturday, June 21,
at The Endocrine Society’s 85th Annual Meeting in Philadelphia. The findings provide
preliminary data for further research into possible therapeutic applications of yoga in modifying
hormone levels in healthy individuals as well as people with specific endocrine disorders.

Yoga has become increasingly popular as a means for coping with stress. To date, there are few
studies investigating effects of classical yoga on hormone regulation. In 1995, research
presented at the The Endocrine Society’s Annual Meeting suggested that classical yoga practices
may have a specific effect on cortisol, a hormone whose levels are strongly associated with states
of stress. In addition to a wide range of physiologic functions in normal everyday activity,
cortisol plays a crucial role in the “fight or flight” stress response. Simply stated, acute and
chronic stress are known to cause elevations in blood and tissue cortisol levels.

Dr. George Brainard, also an author from the 1995 study, and colleagues in Philadelphia sought
to repeat the previous study to verify if a one-hour set of classical yoga practices can lower blood
cortisol levels in healthy female and male vounteers with no past history of practicing yoga. In
addition, they wanted to find out if yoga could lower blood cortisol levels in the same healthy
subjects who have practiced yoga for seven consecutive days.

In this study,16 healthy female and male volunteers were recruited for an eight-day study. On
the first (control) day, blood samples were collected and cortisol levels were measured before
and after a 50-minute quiet period, without the intervention of classical yoga. On the second
day, blood cortisol levels were measured immediately before and after a 50-minute set of
classical yoga practices, guided by an experienced yoga instructor.

Classical yoga practices used during the study included a variety of postures that promote
strength, flexibility and balance of the body as well as massaging and stimulating the internal
organs. For the following five days, subjects were guided through 50-minute sets of yoga
practices with no blood sampling. On the seventh day, blood was sampled and cortisol measured
before and the yoga set. All blood samples were collected at approximately 11 a.m. and noon.
Of the 48 total, paired samples collected during the study, 42 showed a decrease in blood
cortisol, which is consistent with normal daily decreases observed between 11 a.m. and noon.
When compared to the control day, however, larger decreases were observed on the yoga
practice days. These data are consistent with the earlier study and establish evidence of how
yoga influences hormone secretion from the adrenal gland and provides preliminary data for
future studies of how classical yoga influences human biology.

This study was supported by Thomas Jefferson University Center for Integrative Medicine and
The Yoga Research Society.
                                           # # #


                                                186
                                                          Steroid Hormone Action & Metabolism
P3-209
Selective Glucocorticoid Receptor Modulators for the Treatment of Inflammatory Disease.
Jeffrey N Miner*1, Peer Jacobson2, Linda V Kessler1, Christopher Larson1, Junlian Hu1, Debbie Osburn1, Masaki
Nakane2, Chun W Lin2, Jon Rosen1. 1Molec and Cell Biol, Ligand Pharmaceuticals, San Diego, CA; 2Abbott Labs,
Abbott Park, IL.

Glucocorticoids are commonly used to treat inflammatory disease, unfortunately the long-term use of these steroids
leads to a large number of debilitating side effects. The anti-inflammatory effects of glucocorticoids are a result of
GR-mediated inhibition of expression of pro-inflammatory genes as well as GR mediated activation of anti-
inflammatory genes. Similarly, side effects are most likely due to both activated and repressed GR target genes in
affected tissues. An as yet unachieved pharmaceutical goal is the development of a compound capable of separating
detrimental side effects from anti-inflammatory activity. We describe the discovery and characterization of AL-438,
a glucocorticoid receptor ligand that exhibits an altered gene regulation profile, able to repress and activate only a
subset of the genes normally regulated by glucocorticoids. When tested in vivo, AL-438 retains full anti-
inflammatory efficacy and potency comparable to steroids, but has less negative effects on bone metabolism and
glucose control at equivalently anti-inflammatory doses. The mechanism underlying this selective in vitro and in
vivo activity may be the result of differential cofactor recruitment in response to ligand. This compound serves as a
prototype for a unique, nonsteroidal alternative to conventional glucocorticoids in treating inflammatory disease.

BASIC POSTER: Steroid Hormone Action & Metabolism (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                         187
                                                 Steroid Hormone Action & Metabolism
P3-209 News Summary
New compounds have anti-inflammatory effects of steroids without side effects

Researchers have identified compounds that exhibit the anti-inflammatory benefits of steroids
but without the harmful side effects, according to a new study being presented on Saturday, June
21, at The Endocrine Society’s 85th Annual Meeting in Philadelphia. They hope that the
information from this study can be used to further refine these compounds as well as discover
new compounds with improved activity.

Corticosteroids (steroids), such as prednisone or dexamethasone, provide excellent therapeutic
benefit in the treatment of numerous inflammatory diseases. Unfortunately, currently used
steroids cause a variety of serious side effects – such as increased risk for osteoporosis and
fractures, hyperglycemia, hypertension and stunted growth in children – that ultimately limit
their use. Researchers at Ligand Pharmaceuticals and Abbott Laboratories are attempting to
discover new compounds that maintain the anti-inflammatory activity of steroids but lack some
of their side effects.

Steroids exert their effects through the glucocorticoid receptor (GR), a member of a family
known as the nuclear receptors. Activation of the GR by binding to a steroid or other ligand
causes the receptor to regulate the transcription of many genes. Some of these genes are related
to the anti-inflammatory effect of activated GR, and others are associated with its side effects.
To separate these activities, researchers at the two companies, led by Dr. Jeffrey Miner, exploited
differences in the mechanism by which genes involved in the anti-inflammatory effect of steroids
are regulated compared with the regulation of genes that cause some of the side effects.

They established assays that could determine whether one or both sets of genes are regulated.
All tested steroids activated the GR to regulate both sets of genes. They were, however, able to
identify non-steroidal compounds that bound to the GR and regulated the genes involved in
inflammation but had little effect on the genes involved in side effects. In vivo studies
demonstrated the in vivo efficacy of this class of compound combined with reduced effects on
glucose homeostasis, a side effect that can lead to diabetes after long-term steroid treatment. In
addition, these molecules have less impact on bone loss. Therefore, the researchers believe that
they have identified compounds with the efficacy of steroids but reduced side effects.

They then investigated the mechanism of action of these compounds. Recently, a variety of
proteins, called cofactors, have been shown to bind to nuclear receptors. Cofactors involved in
repressing the activity of a receptor, called corepressors, tend to be bound in the absence of
ligand, while cofactors that are required for transcriptional activation, called coactivators,
associate with the receptor after ligand is bound. They showed that a distinct set of cofactors are
bound to the receptor in the presence of steroids but only a subset of those cofactors are
associated in the presence of the nonsteroidal compounds.

The research was supported by Ligand Pharmaceuticals and Abbott Laboratories.
                                          # # #


                                                188
                                                       Steroid Hormone Action & Metabolism
P3-212
Pharmacological Doses of Androgen Do Not Improve Muscle Quality in Young or Older Men: Results from
Two Studies.
Edward T Schroeder*1,2, Michael Terk1,2, Fred R Sattler1,2. 1Keck Sch of Med, Univ of Southern California, Los
Angeles, CA; 2Biokinesiology & Physical Therapy, Univ of Southern California, Los Angeles, CA.

Androgen therapy increases skeletal muscle mass and maximal voluntary strength. However, the effects of
androgens on muscle quality (MQ, strength per unit of muscle) are unknown. Purpose: To determine whether
androgen therapy alone improves MQ in two different populations. Methods: Study #1: 23 HIV+ men
(age=38.7±8.7 years) were randomly assigned to receive nandrolone (600 mg/week) alone or in combination with
resistance training (RT). Study #2: 33 older men (age=72.4±5.2 years) were randomly assigned to receive
oxandrolone (20 mg/day) or placebo. Maximal strength for the leg press, leg extension, and leg flexion were
determined by the 1-repetition maximum (1-RM) method. Muscle cross-sectional area (CSA) of the thigh and lower
extremity lean body mass (LBM) were measured by MRI and DEXA, respectively. MQ by MRI and DEXA were
calculated dividing strength (kg or N) by CSA (cm²) or lower extremity LBM (kg), respectively. Results:
Change (weeks 0-to-12)           Nandrolone Nand+RT Oxandrolone Oxan Placebo
Strength (kg or N)--total thigh 24.5±20.7* 69.3±29.4*        76±79*          0±66
Strength (kg)--quadriceps           6.0±5.4* 28.9±10.9*        N/T            N/T
Strength (kg)--hamstrings           4.8±4.1* 12.9±3.7*      4.4±5.9*        0.2±3.1
Muscle CSA (cm²)--total thigh 15.4±8.0* 15.1±5.5* 11.8±8.5*                 1.4±6.9
Muscle CSA (cm²)--quadriceps 7.2±4.0*          7.2±3.0*     4.7±3.6*       -0.2±3.1
Lower Extremity LBM (kg)            1.6±0.7*   1.7±0.6*     0.9±0.8*       -0.1±0.3
MQMRI (kg/cm²)--total thigh        0.04±0.13 0.38±0.19* -0.2±0.6            0.0±0.5
MQMRI(kg/cm²)--quadriceps         -0.01±0.11 0.35±0.18*        N/T            N/T
MQDEXA(kg/kg)--total thigh         0.02±0.02 0.04±0.02*      0.5±5.6        0.7±3.7
MQDEXA (kg/kg)--hamstrings           3.2±2.6   9.1±4.6* -0.03±0.10        -0.02±0.08
Means±SD; N=Newtons; N/T=not tested; *significant within group change p<0.003
Conclusion: Pharmacological doses of androgen increased thigh muscle CSA, lower extremity LBM, and maximum
strength in both groups but gains were modest. Despite these adaptations and adequate statistical power to show a
15% change in MQ, MQ assessed by MRI or DEXA did not improve as gains in strength were proportional to gains
in muscle mass. The addition of RT to androgen therapy improved MQ in subjects with HIV since the gains in
strength were disproportionately greater than gains in muscle mass.
          Supported by NIH Grants DK-49308 and NCRR GCRC MOI RR-43[br]Bio Technology General Corp.

BASIC POSTER: Steroid Hormone Action & Metabolism (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                      189
                                                Steroid Hormone Action & Metabolism
P3-212 News Summary
Weight training with male hormone therapy can improve muscle quality in important
patient groups

Resistance training appears to be a necessary strategy to optimize muscular adaptations to male
hormone therapy – called androgens, according to studies of HIV positive and elderly men being
presented on Saturday, June 21, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

Androgen therapy increases muscle mass and strength; however, the effects of androgens on
muscle quality – strength per given amount of muscle – are unknown. Increases in muscle
quality would be optimal because gains in strength would be proportionally larger than the
increases in muscle mass, as occurs with resistance training.

To determine if androgen therapy improves muscle quality, Dr. E. Todd Schroeder and
colleagues in Los Angeles conducted two different studies in two very different populations. In
the first study, 23 HIV positive men, with an average age of 39, were randomly assigned to
receive nandrolone alone or in combination with resistance training.

In the second study, 33 older men, with an average age of 72, were randomly assigned to receive
oxandrolone or placebo. To evaluate muscle quality, maximal strength for the leg press, leg
extension and leg flexion exercises were determined. Muscle cross-sectional area (CSA) of the
thigh and lower extremity lean body mass (LBM) were measured by MRI and DEXA,
respectively. Muscle quality by MRI and DEXA were calculated dividing strength by CSA or
lower extremity LBM, respectively.

Treatment with androgens increased muscle CSA, lower extremity LBM and strength in both
populations, which are important adaptations contributing to improved physical function and
health. However, despite these adaptations, muscle quality did not improve, as gains in strength
were proportional to gains in muscle mass. The addition of resistance training to androgen
therapy improved muscle quality in patients with HIV because the gains in strength were
disproportionately greater than gains in muscle mass.

This study was supported by the National Institues of Health and Bio Technology General Corp.

                                             # # #




                                              190
  Steroid Receptors & Steroid Hormone Action in the Cardiovascular System
P1-82
A Variant Form of the Glucocoticoid Receptor Is Translated from a Transcipt Containing an Open Reading
Frame of Exon 1B.
Francine Lafaille1, Kazunori Suzuki1, Tony Antakly*1. 1Biochem, Univ of Montreal, Montreal, Canada.

Accumulating data show that members of the steroid receptor superfamily could be expressed as alternate forms
which may reflect tissular and functional specificity. The most recent analysis of rat glucocorticoid receptor (GR)
gene organization reveals a more complex structure than suspected earlier. At least eleven variants of the 5'
untranslated exon 1
and several alternate promoters could exist. These exon 1 variants are annexed to the main GR gene transcript to
form transcripts differing only within the 5'-termini. Exon 110 sequence analysis reveals a putative open reading
frame (ORF). If translated, this ORF could code for a 40 amino acids peptide that may have physiological relevance.
We developed an antibody (Ab 60) against this ORF peptide. Extensive characterization of Ab 60 proved its
monospecificity. We used Ab 60 as a probe in experiments based on RIA, Western blots, immunoprecipitation and
immunocytochemistry at the EM level. As seen in Western blots, Ab 60 immunoreacted with one major 98 ±5 kDa
protein band in rat liver cells. The molecular weight of this form is slightly higher than the classical GR (93 ±5
kDa) detectable within the same extract. The EM immunocytochemistry using gold-labeled probes also confirms
exon 110 ORF recognition by Ab60. Depending on the tissue cell
types, expression of this ORF differs in magnitude and in its distribution between the cytoplasm, mitochondria and
the nucleus. Heterochromatin, unlike the euchromatin, shows prominent immunoreactivity in several tissues.
Overall, our data establish that this ORF peptide is indeed expressed. Furthermore, its subcellular distribution is
different from the well known "nuclear GR". Since a stop codon is located at the end of the ORF, it is
evident that the translational machinery must ignore it in order to generate the large 98 kDdal protein. Sincesuch a
readthrough process in vertebrates is extremely rare, this study sets a precedent. The functional role of this novel
GR forms is currently under inestigation.

BASIC POSTER: Steroid Receptors & Steroid Hormone Action in the Cardiovascular System (11:00 AM - 12:00
PM and 2:30 PM - 3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                       191
   Steroid Receptors & Steroid Hormone Action in the Cardiovascular System

P1-82 News Summary
Precedent-setting study lays groundwork for therapy of steroid-related human diseases

A novel molecular form of the cortisol receptor is produced in animal and human cells by a
mechanism “borrowed” from unicellular organisms like viruses and bacteria, according to a
study being presented on Thursday, June 19, at The Endocrine Society’s 85th Annual Meeting in
Philadelphia. These results, say the researchers, signify that the cell can inactivate physiological
responses to a hormone by physically sequestering its receptor away from active genes.

In higher organisms, steroid hormones regulate a wide variety of physiological functions, such as
stress, immunity, reproduction and response to disease. These actions are mediated by hormone
receptors within the nucleus of target cells like the brain, lymphocytes and endocrine tissues. It
is widely accepted that steroid receptors elicit their action by binding to active nuclear genes;
however, the current study unveils another mechanism for their action outside these sites.
Researchers at the University of Montreal, led by Dr. Tony Antakly, discovered a novel form of
the cortisol receptor, which is expressed by human and rat cells through an unusual mechanism.
This novel form of the receptor targets not the classical active nuclear genes found in the loose
chromatin, but it results in the accumulation of the receptors in the cytoplasm and on the
condensed chromatin where inactive genes are kept. Sequestering the receptors away from
transcriptionally active chromatin prevents them from regulating nuclear gene expression.

Exon 110 sequence analysis reveals a putative open reading frame (ORF). If translated, this ORF
could code for a 40 amino acids peptide. The researchers developed and characterized an
antiserum (Ab 60) against this ORF peptide. Extensive characterization of Ab 60 proved its
monospecificity. They used Ab 60 as a probe in experiments based on immmunoassay, Western
blots, immunoprecipitation and immunocytochemistry at the Electron Microscope level.
Overall, their data establish that this ORF peptide is expressed. Furthermore, its subcellular
distribution is different from the well-known “nuclear GR.” Because a stop codon is located at
the end of the ORF, it is evident that the translational machinery must ignore it in order to
generate a large 98 kDa protein. The rat GR exon 110 open reading frame is ended by a stop
codon (UGA) and is separated by 17 base pairs from the GR exon 2 start codon. Two molecular
mechanisms of translation regulation are required for the expression of this putative protein,
including preventing the release of the translation complex when it reaches the stop codon and a
jump of the ribosomal translation complex is required in order to “readthrough.”

This mechanism is commonly found in viruses and bacteria, where a distortion of the mRNA
structure allows different proteins to be translated from the same gene. This readthrough process
in humans is extremely rare; therefore, this study sets a precedent. Conceptually, this finding has
important clinical implications. In fact, if researchers could block this readthrough process using
drugs that affect ribosomal functioning, then normal physiological responses could be expected.

This study was supported by the Canadian Institutes of Health Research.
                                            # # #


                                                192
                                                                         Steroid Hormone Biosynthesis
OR23-6
Unexpected Phenotype of P450c17 Knockout Mice.
Synthia H Mellon*1, Susanna R Bair1. 1Ob, Gyn & Reprod Scis, Univ of California, San Francisco, San Francisco,
CA.

Neurosteroids, steroids that are made in and act on cells in the nervous system, are synthesized in a
developmentally-, regionally-, and cell-specific fashion in species from frogs to human beings. Neurosteroids act as
ligands for ion-gated neurotransmitter receptors such as GABAA or NMDA, and augment the effects of GABA and
NMDA at those receptors. Our studies on the developmental function of the neurosteroid dehydroepiandrosterone
(DHEA) and DHEA-sulfate (DHEAS) indicates that they are distinct neurosteroids: DHEA stimulates axonal
growth from specific populations of neocortical neurons while DHEAS stimulates dendritic growth from those cells.
DHEA, but not DHEAS, also affects differentiation of hindbrain cholinergic neurons. P450scc initiates
(neuro)steroidogenesis, converting cholesterol to pregnenolone. Pregnenolone conversion to 17
hydroxypregnenolone and then to DHEA, is mediated by P450c17, an enzyme with both 17 hydroxylase and
c17,20 lyase activities. P450c17 is expressed in specific neurons in the developing rodent nervous system, but does
not appear to be expressed in the adult rodent brain, suggesting that DHEA may have specific functions during
development. To assist us with determining crucial functions of DHEA during neural development, we ablated the
mouse P450c17 gene in 129/SvJ mice. Two lines of targeted ES cells yielded several lines of mice that had
germline transmission of the knockout allele. In all mouse lines, P450c17 -/- mice were not viable, and died during
early gestation, prior to gastrulation and placental growth. The cause of the early lethality is unknown since there is
no known function of fetal steroids at embryonic day 7, the latest time when P450c17-/- embryos were detected.
Heterozygotes were phenotypically normal. Surprisingly, P450c17 is expressed in embryonic endodermal cells in
E7 wild type and heterozygous embryos, but its function in those cells is unknown. Embryos that were not
immunopositively stained for P450c17 were collected by laser catapult microdissection, and DNA was genotyped to
ensure that those embryos were P450c17-/-. Treatment of pregnant females (hetXhet) with subcutaneous pellets
releasing DHEA at a constant rate, or giving pregnant mice DHEA or androstenedione in drinking water all failed to
rescue P450c17-/- mice. Our results suggest that steroid products of P450c17 activity have heretofore unknown
essential functions in early embryonic development.
          Supported by NIH, March of Dimes

BASIC ORAL: Steroid Hormone Biosynthesis (1:00 PM - 2:30 PM)

Presentation Date: 6/21/2003, Time: 2:15 PM; Location: 103 B




                                                         193
                                                             Steroid Hormone Biosynthesis
OR23-6 News Summary
First-time finding shows that enzyme is crucial part of early development of embryo in
mice, possibly humans

An enzyme involved in steroid hormone production – in mice and possibly in humans –has been
shown for the first time to be a crucial part of the early formation and development of an
embryo, according to a new study being presented on Saturday, June 21, at The Endocrine
Society’s 85th Annual Meeting in Philadelphia.

P450c17 is an enzyme involved in steroid hormone production. It converts the steroid
pregnenolone or progesterone into precursors of androgens and estrogens, or into other steroids
called “neurosteroids,” steroids that are active in the brain. Therefore, this enzyme is necessary
for proper reproductive function and for neurological function – in mice and possibly in human
beings.

To assess its role in neurosteroid synthesis, action and development of the mouse nervous
system, Drs. Synthia H. Mellon and Susanna R Bair of the University of California, San
Francisco, removed the P450c17 gene in transgenic mice. They obtained several lines of mice,
and the knockout mice were not viable and were not born.

The researchers demonstrated that they died very early in gestation, approximately embryonic
day seven. They found that P450c17 is expressed in the embryonic endoderm at that time. The
researchers tried to rescue P450c17 knockout mice by treating pregnant moms with various
steroids that they thought might be missing in the embryos, but no steroid treatment was
successful. This study, say Drs. Mellon and Bair, provides the first evident of functions of
P450c17 in early embryogenesis.

This work was supported by the March of Dimes, the National Science Foundation, and the
National Institutes of Health.

                                              # # #




                                                194
Thyroid
                                   Hyperthyroidism, Thyroid Nodules & Thyroid Cancer
P1-506
Primary Prevention of Thyroid-Associated Ophthalmopathy by Pentoxifylline.
Csaba Balazs1, Nadir R Farid*2. 1III Dept of Med, Kenezy Teaching Hosp of Debrecen, Debrecen, Hungary;
2
  Osancor Biotech Inc, Watford, Herts, United Kingdom.

Thyroid associated opthalmopathy (TAO) is a troublesome manifestation of Graves’ disease (GD) which not
infrequently is difficult to manage. Ever minor degrees of eye involvement may impact quality of life negatively .
We have previously demonstrated that pentoxifylline (PTX) can reverse mild to moderate severe TAO. PTX exerts
its action by inhibiting the elaboration of cytokines by orbit-infiltrating T lymphocytes, thus abrogating the
proliferation of orbital fibroblasts, their synthesis of glycosaminoglycans and progression to adipocytes. We
wondered whether PTX would prevent the onset and /progression of TAO if instituted early in course of GD &
whether PTX treatment can counteract the predilection to TAO associated with smoking. To this end, we assigned
182 newly diagnosed patients with GD to either treatment with methimazole plus placebo (Group A) or
methimazole plus PTX at a dose of 400 mgs tid ( Group B) for a period of 12 months. Patients with severe TAO at
the time of presentation were excluded from the study. None of the patients received previous radioiodine treatment.
Group A comprised 70 patients (53 F), mean age 44.1 yrs± 15.3 ; 8 patients had mild TAO at recruitment. With 112
patients, mean age 46.8yrs± 7.7 Group B contained 97 females and 13 with mild TAO. Pre-determined variables
were smoking, family history of GD and concurrent endocrine disorders. All patients were euthyroid by the end of
the study. There were proptionately more patients in remission in group B than A [p<0.05]. Smoking presented a
major risk factor for TAO ( OR=7.9), which was further increased by family history of GD ( OR= 9.1 ). Treatment
with PTX was associated with a reduction in the incidence of TAO, in smokers without (OR=2.5) or with a family
history of GD ( OR=2.0,p<0.001 for both).
PTX, a cheap medicament relatively free of side-effects, is efficacious in the primary prevention TAO, irrespective
of smoking. Although the study was not designed to address the influence of PTX on GD remission, it is apparent
that PTX increases its likelihood in spite of smoking. It is likely that cytokines modulated by PTX are important not
only in TAO but also in the maintenance of GD activity in spite of anti-thyroid medications. A strong argument is
mounted to add PTX to thyrostatic therapy of GD, particularly in those who smoke.

CLINICAL POSTER: Hyperthyroidism, Thyroid Nodules & Thyroid Cancer (11:00 AM - 12:00 PM and 2:30 PM -
3:30 PM)

Presentation Date: Thursday, June 19, 2003




                                                        195
                              Hyperthyroidism, Thyroid Nodules & Thyroid Cancer

P1-506 News Summary
Inexpensive drug for blood circulation found to prevent thyroid-related eye disease

Pentoxifylline (PTX) – commonly used to improve blood flow in patients with circulation
problems – significantly reduces the rates of thyroid-associated eye disease, even in patients who
smoke, many of whom would have, otherwise, developed eye disease, according to a new study
being presented on Thursday, June 19, at The Endocrine Society’s 85th Annual Meeting in
Philadelphia.

Graves’ disease is the most common cause for overactive thyroid, called hyperthyroidism. Eye
disease, one of the manifestations of Graves’ disease, frequently presents a challenge in
treatment. In addition, smoking increases the chances of thyroid-associated eye disease several
fold. Researchers from the current study have previously shown that smoking makes Graves’
hyperthyroidism less responsive to treatment. In a preliminary, uncontrolled study, they found
(PTX) to be an effective therapy for mild to moderately severe thyroid-associated eye disease.

These researchers – from Hungary and the United Kingdom, led by senior investigator Dr. Nadir
R Farid – designed a prospective control study to find whether PTX can prevent the onset or
progression of thyroid-associated eye disease when given alongside conventional anti-thyroid
drugs, upon the initial visit with the endocrinologist .

In addition to finding the benefits of PTX to prevent thyroid-associated eye disease, they also
found that hyperthyroidism in the PTX treatment group was more likely to remit after
completing a one-year course of anti-thyroid medication compared to those not receiving PTX.

PTX is inexpensive, relatively free of side-effects, which makes it a good choice for future use
along-side conventional treatment, recommend the researchers.

The research was supported by the Hungarian Ministry of Health and Osancor Biotech Inc.

                                              # # #




                                               196
               Hypothyroidism, Autoimmunity, Goiter & Other Thyroid Disorders
P3-675
Effect of Seaweed Ingestion on Thyroid Function in Postmenopausal Women.
Jane Teas1, Sam Pino2, Thomas G Hurley1, Lewis E Braverman*2. 1Dept of Hlth Promotion, Educ and Behav,
Norman J. Arnold Sch of Public Hlth, Univ of South Carolina, Columbia, SC; 2Dept of Med, Sect of Endocrinology,
Diabetes and Nutrit, Boston Med Ctr, Boston, MA.

We have previously reported that there was a significant (p< 0.007) delay in the development of DMBA-induced
mammary tumors in seaweed treated rats compared to control rats. Breast cancer rates are significantly higher in the
US than in Asia where seaweed is a staple in the Asian diet. Prior to determining whether seaweed ingestion could
possibly affect the occurrence of breast cancer in US women, a study was carried out to determine the effect of the
ingestion of iodine (I) containing seaweed in healthy, postmenopausal American women. Commercially available
samples of 12 different kinds of edible seaweeds were analyzed for iodine content and found to range from 16 ug/g
to 8165 ug/g. We recruited twenty-five women (mean age 58 yr) to a double-blinded randomized crossover study.
Five gm/d of encapsulated seaweed (Alaria esculenta) containing 550 mg I or placebo capsules were given to the
women as follows: 13 women seaweed (A) and 12 women placebo (B) for 6 weeks, a 3 week washout period,
followed by group A placebo and group B seaweed. Thus, 25 women received seaweed or placebo for 6 weeks.
Urine I and serum T4, FTI, TT3 and TSH concentrations were compared at the termination of each 6 week course of
treatment. Urinary I concentrations increased significantly to 568 ± 32 ug/day (mean ± SE) in the seaweed-treated
women compared to 266 ± 32 ug/day on placebo (p<0.0001). There were no significant changes in serum T4, FTI,
and TT3 concentrations during seaweed or placebo ingestion. However, seaweed ingestion induced a significant
rise in the serum TSH concentrations due to the excess I (1.69 ± 0.22 on seaweed vs 2.19 ± 0.22 µU/ml on placebo,
p < .0001). Conclusion: Ingestion of seaweed containing I induced small but significant increases in serum TSH
concentrations in healthy, postmenopausal women. Whether further abnormalities of thyroid function would occur
during more prolonged ingestion of I rich nutrients remains unclear.

CLINICAL POSTER: Hypothyroidism, Autoimmunity, Goiter & Other Thyroid Disorders (11:00 AM - 12:00 PM
and 2:30 PM - 3:30 PM)

Presentation Date: Saturday, June 21, 2003




                                                       197
            Hypothyroidism, Autoimmunity, Goiter & Other Thyroid Disorders

P3-675 News Summary
Long-term intake of seaweed may contribute to thyroid problems in some

The chronic ingestion of seaweed – recommended as an alternative treatment for an underactive
thyroid, or hypothyroidism, by some – may actually contribute to hypothyroidism, according to a
new study of being presented on Saturday, June 21, at The Endocrine Society’s 85th Annual
Meeting in Philadelphia.

In recent years, dietary supplements have been recommended to prevent and treat a wide variety
of illnesses and to improve general health. Seaweed (i.e., kelp, kombu, dolts) has been
recommended to treat hypothyroidism by a leading alternative medicine physician in one of his
monthly publications. Seaweed is rich in iodine, a necessary constituent of the thyroid
hormones. The content of iodine in edible seaweed is extremely variable, ranging from 16 µg
iodine to 8165 µg per gram.

There are previous reports that women and men with normal thyroid function (euthyroid) with
autoimmune thyroid disease (Hashimoto’s thyroiditis) are inordinately sensitive to excess iodine
ingestion and may develop hypothyroidism. Thus, excess iodine is contraindicated in people
with Hashimoto’s thyroiditis, present in approximately 25 percent of U.S. women over age 60.

Dr. Lewis E. Braverman and colleagues have previously reported that seaweed ingestion
prevents the development of breast cancer in rats and the incidence of breast cancer is
significantly lower in Asia where seaweed is a staple in the diet. Prior to determining whether
excess seaweed ingestion could possibly decrease the prevalence of breast cancer in American
post-menopausal women, Dr. Braverman and colleagues in in Boston and South Carolina studied
the potential effects of seaweed ingestion on thyroid function in 25 euthyroid women who did
not have Hashimoto’s thyroiditis. The daily ingestion of 5 gm encapsulated powdered seaweed
(550 µg iodine) for six weeks resulted in a 30-percent increase in serum thyroid stimulating
hormone (TSH) – the first marker of decreased thyroid function – compared to taking a placebo
capsule.

Thus, the chronic ingestion of seaweed might induce hypothyroidism, especially in women with
underlying Hashimoto’s thyroiditis. Furthermore, excess iodine ingestion might induce an
overactive thyroid, or hyperthyroidism, in individuals with thyroid nodules, also common in
older people, especially women. The researchers strongly recommend that commercial seaweed
supplements include the iodine content per serving on the label.

The research was supported by the Susan G. Komen Foundation.

                                             # # #




                                              198
                                                                       Neoplasia of Endocrine Tissues
P2-446
Gene Therapy of Anaplastic Thyroid Carcinoma witha Single Chain Interleukin-12 Fusion Protein.
Yufei Shi1, Ranjit S Parhar2, Minjing Zou1, Essa Baitei1, Ali Alzahrani3, Futwan A Al-Mohana2, Nadir R Farid*4.
1
  Dept of Genet, King Faisal Specialist Hosp & Res Ctr, Riyadh, Saudi Arabia; 2Dept of Biol and Med Res, King
Faisal Specialist Hosp and Res Ctr, Riyadh, Saudi Arabia; 3Dept of Med, King Faisal Specialist Hosp and Res Ctr,
Riyadh, Saudi Arabia; 4Osancor Biotech Inc, Watford, Herts, United Kingdom.

Anaplastic thyroid carcinoma is the most aggressive thyroid malignancy with a mean short survival time . No
effective therapeutic approach is currently available, making the development of novel treatments necessary.
Interleukin 12 (IL-12) is a pro-inflammatory heterodimeric cytokine with strong antitumor activity. In the present
study, we investigated the potential of IL-12 gene therapy for anaplastic thyroid carcinoma in BALB/c (nu/nu ) nude
mice. A single chain IL-12 fusion protein was created to assure equal expression of its p35 and p40 subunits. Human
anaplastic thyroid carcinoma cell line ARO was stably transfected with IL-12 expression plasmid under the control
of CMV promoter (scIL-12/CMV). High levels of functional IL-12 (27± 4.1 ng/ml/10 6 cells/48 h) were produced by
scIL-12 transfected ARO cells (ARO/IL-12). Tumorigenicity in nude mice was completely lost in scIL-12
transfected ARO cells, as demonstrated by the lack of tumor formation following subcutaneous injection of 2 x 106
ARO/IL-12 cells, even though there was no difference in cell proliferation between ARO and ARO/IL-12 cells.
Tumor growth was observed after challenge with ARO tumor cells, indicating that protective immunity had not
developed against the parental cells. Furthermore, the growth rate of established subcutaneous ARO tumors was
significantly reduced by either subcutaneous injection of 2 x 10 6 ARO/IL-12 cells weekly or intramuscular injection
of 50 mg scIL-12/CMV twice weekly. The anti-neoplastic activity was , however, abrogated by intra-peritoneal
injection of anti-NK cell antibody. Moreover, significantly higher number of ARO/IL-12 cells and ARO cells were
killed by splenocytes from nude mice previously treated with ARO/IL-12 as compared to those treated with ARO
cells (32% vs 9%, when ARO were used as targets and 43% vs 17% when ARO/IL12 cells ere used as target,
p<0.01 for both) in an in vitro cytotoxicity assay. Again, tumor cell killing was neutralized by the addition of anti-
NK cell antibody in the assay. In conclusion, we have demonstrated successful immunotherapy with a scIL-12
fusion protein against the anaplastic thyroid carcinoma in an in vivo model. NK cells mediate the immune response
against ARO/IL-12 cells. These results may set the stage for clinical application of IL-12 gene therapy for poorly
differentiated thyroid carcinoma.

BASIC POSTER: Neoplasia of Endocrine Tissues (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                        199
                                                             Neoplasia of Endocrine Tissues

P2-446 News Summary
Gene therapy shows promise for untreatable, aggressive form of thyroid cancer

In the future, people who develop an aggressive and deadly form of thyroid cancer, called
anaplastic thyroid carcinoma, may have hope for surviving the disease with gene therapy,
according to a new study being presented on Friday, June 20, at The Endocrine Society’s 85th
Annual Meeting in Philadelphia.

Anaplastic thyroid carcinoma – mostly occurring in people over 60 years old – is a rapidly lethal
disease, whose course is only slightly modified by currently available treatments. Therefore,
researchers believe it to be a logical candidate for innovative gene therapy. To test the
effectiveness of a gene therapy treatment plan, researchers at the King Faisal Specialist Hospital
& Research Centre in Saudia Arabia, in collaboration with Osancor Biotech Inc. in the United
Kingdom, studied mice that lack thymus-dependent immune cells. The thymus is a small organ
located in the upper front portion of the chest, extending from the base of the throat to the front
of the heart. Human tumor cells injected in these mice survive and form solid tumors.

The researchers, led by senior investigator Dr. Nadir R. Farid of the United Kingdom, injected
human anaplastic tumor cells without genetic manipulation or similar cells that were genetically
engineered to carry a “balanced” interleukin-12 (IL-12) gene, made by joining the two subunits
making up IL-12 end-to-end. IL-12 is known to be a potent inducer of cell-mediated
immunological response to tumors.

The results show clearly, say the researchers, that anaplastic thyroid tumor cells engineered with
IL-12 are easily destroyed by immune cells. In addition, when the engineered tumor cells are
injected alongside similar but unmodified tumor cells, the unmodified cells were also completely
destroyed. The effectiveness of IL-12 was such that when the IL-12 gene was injected directly
into muscle, tumor cells also regressed. In contrast to previous studies using IL-12
immunotherapy in non-thyroidal tumors, which incriminated thymus-dependent T cells, the
researchers found in their studies that the immune attack against anaplastic thyroid tumor cells
was mediated by “natural killer cells.” Natural killer cells are not as particular in selecting their
targets as are T cells and are easier to manipulate.

The researchers believe that these results provide a strong foundation to move to human trials of
IL-12 modified anaplastic cells to initiate immune-mediated regression and cure.

The majority of the research project was funded by the King Faisal Specialist Hospital &
Research Centre. The remaining support came from Osancor Biotech Inc.

                                               # # #




                                                200
                                                                              Thyroid Nodules & Cancer
P2-532
A Novel and Promising Therapy for Anaplastic Thyroid Cancers.
Akira Ohtsuru*1, Alexei Podtcheko1, Hiroyuki Namba1, Shunichi Yamashita1. 1Dept of Molec Med, Atomic Bomb
Disease Inst, Nagasaki Univ Grad Sch of Biomed Scis, Nagasaki, Japan.

 Background & Aim We have already reported the limited efficacy of p53 gene therapy for anaplastic thyroid
carcinomas, which is a poor therapeutic response and lead to high mortality. The non-receptor tyrosine kinase, c-
ABL, along with one of p53 downstream targets, is critical in the regulation of the cell cycle and radiation-induced
cell response. To find out the novel therapy for anaplastic thyroid carcinomas, we investigated at first the therapeutic
potential of a c-ABL-specific tyrosine kinase inhibitor, STI571. Methods & Results Western blot analysis
demonstrated high expression levels of c-ABL in anaplastic thyroid cancer cell lines ARO (mutated p53) and FRO
(non-detectable p53), and in the papillary cancer cell line NPA (mutated p53). These cell lines showed marked
inhibition of cell growth after treatment with STI571. In contrast, the growth of TPC-1 and KTC-1, papillary thyroid
cancer cell lines which carry wild type p53 and have low levels of c-ABL, was not inhibited by STI571.
Furthermore, STI571 did not affect the growth of 1F3, a cell line generated from FRO by stable transfection with
wild type p53. Immunohistochemical study has demonstrated that undifferentiated thyroid carcinomas highly
express c-ABL as well as mutant p53 nuclear staining. Colony formation assay showed that STI571 enhanced the
effect of ionizing radiation in anaplastic cancers, but not in differentiated cancer. Cell cycle analysis and PARP
cleavage revealed that STI571 treatment increased the apoptotic fraction of FRO and ARO cells, but not in normal
or differentiated cancer cells harbored with wild type p53. These changes were accompanied by inhibition of c-ABL
phosphorylation /activation and increased expression of p21cip1 in FRO and p27kip1 in both FRO and ARO cells.
The growth of FRO cells implanted into immunocompromised mice was inhibited by STI571 and eradiated by the
STI571 and radiation (10 Gy) combination therapy. In contrast, tumor growth did not changed in 10 Gy radiation
alone compared with untreated control group. Discussion & Conclusion The clinical usefulness of STI571 has
been demonstrated in chronic myelogeneous leukemia and gastrointestinal stromal tumors. In conclusion, this is a
first report that c-ABL is overexpressed in anaplastic thyroid carcinoma cells, and selective suppression of c-ABL
activity by STI571 is a potential molecular targeting therapy in combination with radiation therapy for p53-defective
or -mutated thyroid carcinomas.

CLINICAL POSTER: Thyroid Nodules & Cancer (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                         201
                                                                Thyroid Nodules & Cancer
P2-532 News Summary
Novel leukemia drug may be valuable treatment for rare, deadly form of thyroid cancer

Researchers in Japan have found compelling evidence that a novel drug for treating a type of
leukemia is also a promising therapy for anaplastic thyroid cancers, according to a new study
being presented on Friday, June 20, at The Endocrine Society’s 85th Annual Meeting in
Philadelphia.

Anaplastic thyroid cancer grows very rapidly, is an invasive type of thyroid cancer and responds
poorly to therapy. It occurs most often in people over 60 years old. The cause is unknown, and
thyroid function tests are usually normal. Anaplastic cancer is a very rare disease, accounting for
only about 1 percent of all thyroid cancers. The quality of life of these patient is impaired, and
their prognosis is not good.

A new therapy for metastatic or recurrenced thyroid cancers is highly desirable. Through their
research, Dr. Akira Ohtsuru and colleagues at the Atomic Bomb Disease Institute in Nagasaki,
Japan, have discovered that ST1571, a novel enzyme inhibitor drug for leukemia, is effective to
kill the anaplastic thyroid cancer cells in a cell culture system and a tumor implanted in an
animal model, without evident side effects. Thus, the selective suppression of c-ABL activity by
ST1571 may represent a potential anti-cancer strategy for human anaplastic thyroid carcinomas.

The tyrosine kinase inhibitor, STI571, selectively suppresses the activity of ABL, platelet-
derived growth factor receptor (DGFR) and c-KIT. In chronic myelogenous leukemia (CML),
proliferation and growth of BCR/ABL expressing tumor cells can be blocked by STI571. In
clinical trials, treatment with STI571 led to the remission of CML, with minimal side effects.

                                              # # #




                                               202
                                     The Thyroid & Thyroid Dysfunction: Human Biology
OR10-1
Ultrasound-Guided Percutaneous Ethanol Injection (PEI) in Thyroid Cystic Nodules: A Randomized
Controlled Trial.
Andrea Frasoldati1, Marialaura Pesenti1, Francesca Briganti1, Michele Zini1, Roberto Valcavi*1. 1Unit of
Endocrinology, Arcispedale Santa Maria Nuova, Reggio Emilia, Italy.

Ultrasound (US)-guided PEI for benign thyroid cystic nodules has been proposed as an alternative treatment to
surgery. However, EBM studies on PEI efficacy and safety are still lacking. This may have prevented the spreading
of routine PEI treatment, and surgery remains the first choice in most centers.
Study design. We carried out a prospective controlled study on 259 consecutive patients of both sexes (203 F, 56
M), aged 18-83 years, with benign thyroid cystic nodules. Inclusion criteria were: local discomfort or cosmetic
damage, volume >2 ml , 50% or more fluid component as assessed by US examination, benignity as demonstrated
by cytology obtained by (US)-guided Fine Needle Aspiration Biopsy (FNAB), euthyroidism. Patients with
inadequate, suspicious or positive FNAB cytology, with elevated serum Calcitonin, and laryngeal cord palsy were
excluded. Patients were randomly assigned to receive either simple cyst evacuation (N=127) or evacuation plus
ethanol injection by a skilful operator using a US-guided technique (N=132). Ethanol amount injected corresponded
to 50-70 % of cystic fluid extracted. Patients were then examined after 1, 3, 6, 12 months. If necessary, a second and
a third ethanol injection was administered. The two groups were similar regarding age, sex, nodule volume and
characteristics, discomfort score, thyroid hormone and TSH levels, anti-thyroglobulin and anti-TPO antibodies.
Results. Seventy, 39 and 23 patients received 1, 2 or 3 PEI treatments, respectively. Before treatment, the mean
(±SE) nodule volume was 18.3±1.6 vs 19.9±1.2 in the PEI vs the simple evacuation group (n.s.). After one year,
volumes were 5.4±0.9 vs 16.1±1.2 (p<0.001), with a median 86.7 % vs 6.25 % reduction of the initial volume
(p<0.001). In patients with unicamerated, (>80% fluid component, N=84) median nodule volume reduction after PEI
achieved 92.1%, while in mixed lesions (50-80% fluid component, N=48) was 74.7%. Compressive and cosmetic
symptoms improved/disappeared in 66.6% and 71.9 % of patients treated with PEI vs 29.1% and 25.9% of patients
treated with simple evacuation, respectively (p<0.001). One patient treated with PEI had transient laryngeal
dysfunction, subsided after 2 months. No permanent vocal cord palsy, nor other major side effects occurred.
Conclusions: our data provide definitive evidence that PEI is a safe and effective treatment for cystic thyroid
nodules. Unicamerated thyroid cystic nodules are the most suitable for PEI.

CLINICAL ORAL: The Thyroid & Thyroid Dysfunction: Human Biology (1:00 PM - 2:30 PM)

Presentation Date: 6/19/2003, Time: 1:00 PM; Location: Exhibit Hall C




                                                        203
                               The Thyroid & Thyroid Dysfunction: Human Biology

OR10-1 News Summary
Simple procedure can cure benign thyroid cysts, bypassing need for surgery

A simple interventional procedure is a safe, effective and curative treatment for benign cystic
thyroid nodules, according to a study being presented on Thursday, June 19, at The Endocrine
Society’s 85th Annual Meeting in Philadelphia. The study also showed that an alternative
therapy – aspiration of the fluid content of thyroid cysts – is ineffective because thyroid cysts
usually recur. Researchers say that this procedure may become the first therapeutic option to
effectively treat these cysts, thereby allowing patients to avoid unnecessary surgery.

Thyroid cysts are very common, composing approximately 25 percent of all thyroid nodules.
So far, the only final therapeutical option in patients with large thyroid cystic nodules, which
cause neck lumps and local discomfort or compression, has been surgery.

In the current study, Dr. Roberto Valcavi and colleagues at the Arcispedale Santa Maria Nuova
in Reggio Emilia, Italy, treated 132 adult patients with benign thyroid cysts by ultrasound-guided
percutaneous ethanol injection (PEI), an interventional procedure. These cysts, on average, had
a volume of 18 ml (3–4 cm in diameter). This group of patients was compared with 127 patients
with similar thyroid cysts that were, instead, submitted only to aspiration of the fluid content.

Patients were randomly assigned to the two groups – PEI versus aspiration only. All patients
were re-examined with ultrasound after 12 months. Researchers note that a year interval is
sufficiently long to evaluate whether the treatment can be considered definitive. After one year,
PEI patients had, on average, an 86-percent volume reduction while the simple aspiration group
had only a 6-percent reduction. The results were even better, a 92-percent reduction, considering
a subset of cystic lesions – single/empty body cysts – treated with PEI. Also symptoms of local
compression and/or cosmetic complaint, initially referred by patients, significantly improved or
disappeared in the group treated with PEI. No major side effects occurred.

PEI is performed under ultrasound guidance by inserting a special needle inside the thyroid cyst.
The fluid content is first aspirated and then a small amount of ethanol is injected. The procedure
is controlled in real time by ultrasound imaging. Ethanol administration causes the formation of
scar tissue and, therefore, it is usually a definitive cure, while simple cyst aspiration is not.

As compared to the traditional surgical treatment of large thyroid cysts, the researchers note that
PEI is a valid alternative with several advantages, such as avoids surgical risks, performed in an
outpatient setting, requires no anesthesia (general or local), takes only a few minutes, costs less
and preserves normal thyroid tissue.

The researchers say that this study demonstrates that the clinical management of thyroid cystic
nodules could change in the future. They add that it is up to the patient’s physician to evaluate if
the cystic nodule is suitable for PEI.
                                             # # #


                                                204
                                                                                Autoimmune Endocrine
P2-472
Association between Vitiligo and Thyroid Disfunction: Study of a Large Group of Patients with Vitiligo.
M Appetecchia*1, R Baldelli1, P Iacovelli2, F Diacono1, A Paro Vidolin2, G Leone2. 1Endocrinology, Regina Elena
Cancer Inst, Rome, Italy; 2Dermatol, S. Gallicano Inst, Rome, Italy.

BACKGROUND: Vitiligo affect 1 to 3% of the general population and is characterized by circumscribed
depigmented macules resulting from loss of cutaneous melanocytes. The disease can be associated with autoimmune
disorders such as thyroid diseases, diabetes mellitus, pernicious anemia and alopecia areata. An association between
vitiligo and autoimmune thyroid disease has been reported with a prevalence of up to 30% in vitiligo patients.
Circulating organ-specific autoantibodies to the thyroid are commonly detected in the sera of vitiligo patients. The
aim of this study was to investigate the occurrence of autoimmune thyroid disease in patient with vitiligo.
PATIENTS AND METHODS: 62 patients affected by vitiligo were seen between 2000 and 2002. All patients (50
females and 12 males, age 47.0±13.0 yrs., disease onset of 12.5±12.6 yrs.) received information about the study and
accepted to participate. More frequently the vitiligo occurred around the eyes and mouth, the anterior neck, elbow
and fingers, perineum, palmar wrist and dorsal ankles, anterior tibial regions and low back. A blood sample was
collected for thyroid function evaluation. TSH, FT3, FT4, Thyroglobulin, TPO-Ab and TG-Ab (n.v. TSH 0.3-4.2
mUI/ml, FT3 1.8-4.2 pg/ml, FT4 9.3-17.0 pg/ml, TPO-Ab 0-32 IU/ml , TG-Ab 0-115 IU/ml). TSH values higher or
lower the normal range were considered for hypo- or hyperthyroidism respectively. RESULTS: Thyroid disfunction
was present in 20 out of 62 vitiligo patients; in particular subclinical hyperthyroidism (TSH < 0.3 mUI/ml) was
found in 2 out of 62 patients (3.2%), clinical hypothyroidism (TSH > 4.2 mUI/ml and FT4 < 9.3 pg/ml) in 8 out of
62 patients (13%), subclinical hypothyroidism (TSH > 4.2 mUI/ml) in 10 out of 62 patients (16%). Positive thyroid
antibodies were present in 49 out of 62 patients (79%). Among the 49 patients studied with positive autoantibodies,
61% showed an autoimmune disease with euthyroidism, 4% hyperthyroidism and 35% hypothyroidism. No
relationship was seen between the sites and the number of the lesions at presentation, the average disease duration
and thyroid status. DISCUSSION: Our preliminary data confirm that the occurrence of both clinical and subclinical
autoimmune thyroid diseases is increased in vitiligo patients; subclinical thyroid disease indicates regular
surveillance while the clinical evidence of thyroid disfunction could require an adequate treatment.

CLINICAL POSTER: Autoimmune Endocrine (11:00 AM - 12:00 PM and 2:30 PM - 3:30 PM)

Presentation Date: Friday, June 20, 2003




                                                       205
                                                                      Autoimmune Endocrine
P2-472 News Summary
Skin condition with loss of pigmentation carries high risk of autoimmune thyroid disease

People with a skin condition characterized by smooth, white spots on the skin have a high risk of
developing autoimmune thyroid disease, according to a new study being presented on Friday,
June 20, at The Endocrine Society’s 85th Annual Meeting in Philadelphia.

Depigmented areas of the skin characterize the skin disease called vitiligo. It is a disorder
caused by the presence of autoantibodies against its own skin-related antigens, resulting in an
autoimmune disease that can be accompanied by alterations in the eyes. In addition, specific
autoantibodies against other organs, such as the thyroid, pancreas and blood cells can be present
in patients affected by vitiligo. Autoantibodies against the thyroid can result in different types of
diseases. An association between vitiligo and autoimmune thyroid disease has been reported,
with a prevalence of up to 30 percent in vitiligo patients compared to a 1-percent prevalence in
the general population. The aim of current study, led by Dr. M. Appetecchia, was to investigate
the occurrence of autoimmune thyroid disease in patients with vitiligo treated at the Regina
Elena Cancer Institute in Rome.

Sixty-two patients affected by vitiligo were seen between 2000 and 2002. All patients –
including 50 women and 12 men, 34–60 years old – received information about the study and
accepted to participate. A general history was taken from all patients and a full clinical
examination was carried out. Vitiligo was defined as circumscribed lesions characterized by a
variable number of pale, white macules that tended to enlarge centrifugally. More frequently,
the vitiligo occurred around the eyes and mouth, the anterior neck, elbow and fingers, perineum,
palmar wrist and dorsal ankles, anterior tibial regions and low back. A blood sample was
collected for thyroid function evaluation, including thyroid-stimulating hormone (TSH), free
triiodothyronine (FT3), free thyroxine (FT4), thyroglobulin, TPO and Tg autoantibodies. TSH
values higher or lower the normal range were considered for hypo- or hyperthyroid condition,
respectively.

Autoantibodies against the thyroid were present in a large number of patients, 49 out of 62
patients (79 percent). Among the 49 patients studied with positive autoantibodies, 61 percent
showed an autoimmune disease with euthyroidism, 4 percent with hyperthyroidism and 35
percent with hypothyroidism. No relationship was seen between the sites and the number of the
lesions at presentation, the average disease duration and thyroid status. Thyroid dysfunction was
present in 20 out of 62 vitiligo patients; in particular subclinical thyroid hyperfunction condition
(TSH < 0.3 UI/ml) was found in two out of 62 patients (3.2 percent), clinical thyroid
hypofunction (TSH > 4.2 UI/ml and FT4 < 9.3 pg/ml) in eight out of 62 patients (13 percent),
and subclinical thyroid hypofunction (TSH > 4.2 UI/ml) in 10 out of 62 patients (16 percent).

Preliminary data underline the occurrence of autoantibodies against the thyroid with both clinical
and subclinical thyroid dysfunction in vitiligo patients. Subclinical thyroid disease indicates
regular surveillance while the clinical evidence of thyroid dysfunction could require treatment.
                                               # # #


                                                206
Indices
                      Index of Abstracts by Day of Presentation


Thursday, June 19, 2003

Headline                                                                                   Page #
1. Moderate changes in diet lower levels of a negative marker of heart health                     11
2. Amish lead researchers to possible gene responsible for high blood pressure                    13
3. Form of hypertension is mild in most but shows full characteristics in minorities              33
4. Rare form of hypertension found to be not so uncommon; good implications for treatment 35
5. Heart gene expression is different in the day compared to night; finding important for drug
    development                                                                                   37
6. Diabetic drug shows promise in preventing or treating diabetic kidney disease                  39
7. Sildenafil (Viagra®) improves gastric problem as well as impotence in type-1 diabetics         41
8. Web technology connects diabetes patients with doctor for blood sugar monitoring               43
9. Levels of a hormone can predict diabetics at risk for unhealthy weight gain                    45
10. Large percentage of Asian Indian men in U.S. have abnormal blood sugar, leading to
    diabetes and heart disease                                                                    47
11. First-time finding opens pathway to future diabetes treatment with American ginseng           49
12. First-time finding shows that obese people likely have molecular abnormalities in brain       85
13. Early research identifies potential drug targets for treating obesity and diabetes            95
14. Hormone administered by nose drops reduced food intake, caused weight loss in animals 111
15. Researchers uncover the mechanism of temporary bone loss in breastfeeding women; may
    lead to treatments for osteoporosis                                                           133
16. Smaller-than-expected babies, regardless of prematurity, have higher risk of adult diabetes 135
17. Contrary to previous research findings, male baldness is not a sign of heart disease risk     175
18. Sexual activity after taking sildenafil (Viagra®) may not be heart safe in some diabetic men 177
19. By-product of naturally occurring estrogen may help male hormone–related diseases             179
20. Precedent-setting study lays groundwork for therapy of steroid-related human diseases         191
21. Inexpensive drug for blood circulation found to prevent thyroid-related eye disease           195
22. Simple procedure can cure benign thyroid cysts, bypassing need for surgery                    203


Friday, June 20, 2003

Headline                                                                             Page #
1. Improving blood pressure in diabetics reduces other health problems                      1
2. Thyroid hormone T3 provides more problems than benefits in treating depression, despite
    current treatment guidelines for its use                                                3
3. Growth hormone and testosterone together may counteract effects of aging on the body     5
4. Amino acid found to improve muscle mass in men, possibly leading to aging treatments     7
5. Symptoms of hypogonadism are not related to testosterone deficiency, as many believe     9


                                              207
Headline (Friday, cont.)                                                                  Page #
6. Newly found hormones may hold keys to protect against heart damage                            29
7. Arthritis drug shows promise in preventing sudden cardiac death after heart attack            31
8. Hormones made in the heart may be future treatment for most deadly form of cancer             55
9. Specific proteins that regulate genes may improve diagnosis and treatment of breast cancer
    in the future                                                                                57
10. Unbalanced diets in pregnant women may affect lifelong health of their children              61
11. Growth hormone helps obese people lose weight and improves cholesterol levels                65
12. Growth hormone use in children does not increase risk of cancer                              67
13. Lifelong use of growth hormone does not appear to increase risk of cancer                    69
14. Growth hormone treatment shows lower risk of cancer and heart problems for people with
    pituitary hormone deficiency, strokes still an issue                                         71
15. New family of drugs shows success in preventing atherosclerosis in early studies             79
16. Osteoporosis drug shows rapid and sustained ability to prevent fractures                     113
17. Adults who had heart transplants as children should be checked for osteoporosis              115
18. Menopausal estrogen loss affects risk of osteoporosis more than aging-related bone loss      117
19. Elderly women significantly reduce risk of bone fracture with osteoporosis drug              119
20. Team approach to evaluating fractures leads to better treatment for osteoporosis             121
21. Bone density represents small portion of osteoporosis drug’s effect on preventing fractures 123
22. Calcium deficiency in the urine identifies women at risk for bone loss and fractures         127
23. Vitamin D deficiency is common among adolescents, affecting lifelong bone strength           129
24. Too much and too little vitamin A is bad for bone strength, with increased fracture risk     131
25. Diabetes medicine shown to help adolescents with hormone problem and prevent diabetes 137
26. Diabetic drug shown, for the first time, to prevent endocrine condition in young women       139
27. Long-term steroid treatment in children affects bone strength                                141
28. Smoking During Pregnancy Increases Infants Risk of Heart Disease Later in Life               143
29. Blood vessel function is impaired, leading to heart disease, in women with common
    endocrine problem                                                                            153
30. Testosterone spray shown to increase testosterone to healthy levels in certain women         155
31. Estrogen therapy improves cognitive processes in verbal learning and memory                  157
32. Estrogen found in Chinese healing herb, among other natural products                         159
33. High levels of testosterone in postmenopausal women may be culprit of increased heart
    disease risk                                                                                 161
34. Women’s sleep is better in quantity and quality than men’s, may stem from adaptation         163
35. Combination of male and female hormones are viable male contraceptive                        169
36. Men with hormone disorder respond well to testosterone gel, regardless of testosterone
    levels at treatment                                                                          171
37. Sustained-released, testosterone male contraceptive appears safe, effective for future use   173
38. Novel protein shown to affect heart muscle growth; may hold keys to future heart therapy 181
39. Gene therapy shows promise for untreatable, aggressive form of thyroid cancer                199
40. Novel leukemia drug may be valuable treatment for rare, deadly form of thyroid cancer        201
41. Skin condition with loss of pigmentation carries high risk of autoimmune thyroid disease 205


                                              208
Saturday, June 21, 2003

Headline                                                                                 Page #
1. Testosterone deficiency a problem in older obese men                                          15
2. Endocrine problem in women reduces quality of life and sexual satisfaction                    17
3. Large proportion of men with heart disease have testosterone deficiency; higher in diabetics 19
4. Ultrasound not a reliable in diagnosing specific syndromes marked by ovarian cysts            21
5. Adolescent weight gain and genetic markers identify early maturing girls with likelihood of
    endocrine problem                                                                            23
6. Genetic factor identifies men on testosterone therapy who may develop enlarged prostate 25
7. Derivative of aspirin improves insulin resistance in diabetics                                51
8. Testosterone added to standard HRT may reduce breast cancer                                   53
9. Anti-inflammatory glucocoritcoids increase a person’s risk for heart disease                  63
10. Growth hormone test is not reliable measure in overweight and obese                          73
11. New drug shows improvements over existing therapy to treat life-shortening disease           75
12. Improved method for detecting growth hormone use by athletes may put stop to abuse           77
13. Hormone that controls appetite has varying effects in anorexia, bulimia and obesity          87
14. Low-fat dinner in young, overweight girls promotes the body’s use of fat                     89
15. High levels of a fat cell hormone is linked to risk of stroke in obese men                   91
16. Obese African Americans, especially women, have a high risk of vitamin D deficiency          93
17. Weight-loss and diabetic drugs together helped children lose weight, reduce diabetes risk 97
18. Combining the weight-loss power of two drugs does not add up                                 99
19. Newly found hormone released by stomach may serve as basis for future obesity therapy 101
20. Sleep restriction is linked to weight gain and risk for obesity                              103
21. Early study shows promise for creating weight-loss drugs based on hormone                    105
22. First-time finding shows fat tissue produces a hormone in obese and possibly links it to fat
    cell production                                                                              109
23. Men with severe infertility likely to have genetic defects; common in vitro procedure may
    pass bad genes to offspring                                                                  149
24. Testosterone treatment jump starts effects of Viagra in certain men                          151
25. Soy infant formula may negatively affect mammary gland growth, based on animal study 165
26. New class of non-steroidal drugs may be superior treatment in common prostate problem 183
27. Yoga reduces levels of stress-related hormone                                                185
28. New compounds have anti-inflammatory effects of steroids without side effects                187
29. Weight training with male hormone therapy can improve muscle quality in important
    patient groups                                                                               189
30. First-time finding shows that enzyme is crucial part of early development of embryo in
    mice, possibly humans                                                                        193
31. Long-term intake of seaweed may contribute to thyroid problems in some                       197




                                              209
Sunday, June 22, 2003

Headline                                                                                Page #
1. Gene therapy shows promise for protecting heart muscle from injury in diabetics             27
2. Salivary glands may have role in gene therapy for inherited endocrine problems              59
3. New research project focuses on receptors involved in obesity, diabetes, cancer and aging 81
4. Discovery of the function of genetic flaw in diabetes may lead to new drug treatments       83
5. Brain peptides affect activity level and metabolism, leading to decreased weight            107
6. Long-term bone strength improvements can be predicted by early markers of change
    produced by new osteoporosis drug                                                          125
7. Premature children have metabolic abnormalities that may lead to adult diseases             145
8. Children with gene alteration are more prone to obesity and precursors of diabetes          147
9. First-time finding of mechanics of labor may lead to prevention of premature births         167




                                             210
                               Index of Abstracts by Author


Contact/Author                                   Page #
Albert, Stewart G.                               65
Aldhahi, Waleed A.                               51
Allan, Carolyn A.                                15
Anderson, Robert J.                              1
Antakly, Tony                                    191
Appetecchia, M.                                  205
Artaza, Jorge N.                                 181
Aversa, Antonio                                  151
Aylwin, Simon J.B.                               101
Bazaes, Rodrigo A.                               135
Bevilacqua, Maurizio                             127
Bondy, Carolyn A.                                53
Bonert, Vivien S.                                73
Brar, Bhawanjit K.                               29
Braverman, Lewis E.                              197
Bridger, Tracey                                  137
Ceda, Gian Paolo                                 7
Chines, A                                        113
Christ-Crain, Mirjam                             9
Cohen, Adi                                       115
Colman, Ricki J.                                 11
Condon, Jennifer C.                              167
Coviello, Andrea                                 173
Culler, Michael D.                               105
Dalton, James T.                                 183
DelParigi, Angelo                                85
Destefanis, Silvia                               87
Diamanti-Kandarakis, Evantia                     153
Dobnig, Harald                                   125
Duda, Karen M.                                   83
Fardella, Carlos E.                              33
Farid, Nadir R.                                  195, 199
Ferlin, Alberto                                  149
Fliers, Eric                                     3
Giannoulis, Manthos                              5


                                           211
Contact/Author                       Page #
Gordon, Catherine M.                 129
Hahn, Susanne                        17
Heald, Adrian H.                     45
Hindmarsh, Peter C.                  143
Hofman, Paul L.                      145
Hugo, Eric                           109
Humberstone, Andrew J.               155
Ibanez, Lourdes                      139
Jacobsen, Britta M.                  57
Jaursch-Hancke, Cornelia             177
Joffe, Hadine                        157
Klein, Karen O.                      159
Lansang, M. Cecilia                  39
Lee, Ka Kui                          169
Lin, Shuo-Yen J.                     37
Lowe, Hyesoo                         161
Lukacs, Jane                         117
Lund, Trent D.                       179
Luo, LuGuang                         49
Maffeis, Claudio                     89
Mager, Justin                        185
Makitie, Outi                        141
Malkin, Christopher J.               19
Margolis, Ronald N.                  81
McClung, Michael R.                  119
Mellon, Synthia H.                   193
Miner, Jeffrey N.                    187
Ohtsuru, Akira                       201
Olsson, Tommy                        91
Ong, Huy                             79
Opotowsky, Alexander R.              131
Padilla-Banks, Elizabeth             165
Parikh, Shamik J.                    93
Penev, Plamen                        103
Phillips, David I.W.                 61
Pignatelli, Duarte                   21
Piswanger-Soelkner, Jutta C.         41



                               212
Contact/Author                     Page #
Powell, David R.                   95
Rutherford, Sue                    13
Schroeder, Edward T.               189
Schulz, Carla                      111
Segal-Lieberman, Gabriella         107
Shenkerman, Galina                 99
Simon, Nicole                      47
Stowasser, Michael                 35
Streeten, Elizabeth A.             121
Svensson, Johan                    71
Toorians, Arno W.F.T.              175
Trainer, Peter J.                  75
Valcavi, Roberto                   203
VanHouten, Joshua                  133
Vesely, Brian A.                   55
Vgontzas, Alexandros N.            163
Vigersky, Robert A.                43
Voutetakis, Antony                 59
Walker, Brian R.                   63
Wang, Christina                    171
Wang, Ping H.                      27
Watts, Nelson B.                   123
Wilton, Patrick                    67, 69
Witchel, Selma F.                  23
Wu, Zida                           77
Yanovski, Jack                     147
Yu, Xichun                         31
Ziai, Fuad                         97
Zitzmann, Michael                  25




                             213
                 Index of Abstracts by Subject and Headline

1. Clinical Trials/Hot Topics

Headline                                                                                       Page #
Improving blood pressure in diabetics reduces other health problems                            1
Thyroid hormone T3 provides more problems than benefits in treating depression,
despite current treatment guidelines for its use                                               3
Growth hormone and testosterone together may counteract effects of aging on the body           5


2. Aging

Headline                                                                                       Page #
Amino acid found to improve muscle mass in men, possibly leading to aging treatments           7
Symptoms of hypogonadism are not related to testosterone deficiency, as many believe           9
Moderate changes in diet lower levels of a negative marker of heart health                     11
Amish lead researchers to possible gene responsible for high blood pressure                    13


3. Androgen Disorders

Headline                                                                                       Page #
Testosterone deficiency a problem in older obese men                                           15
Endocrine problem in women reduces quality of life and sexual satisfaction                     17
Large proportion of men with heart disease have testosterone deficiency; higher in diabetics   19
Ultrasound not a reliable in diagnosing specific syndromes marked by ovarian cysts             21
Adolescent weight gain and genetic markers identify early maturing girls with likelihood of
    endocrine problem                                                                          23
Genetic factor identifies men on testosterone therapy who may develop enlarged prostate        25


4. Cardiovascular Endocrinology

Headline                                                                                       Page #
Gene therapy shows promise for protecting heart muscle from injury in diabetics                27
Newly found hormones may hold keys to protect against heart damage                             29
Arthritis drug shows promise in preventing sudden cardiac death after heart attack             31
Form of hypertension is mild in most but shows full characteristics in minorities              33
Rare form of hypertension found to be not so uncommon; good implications for treatment         35
Heart gene expression is different in the day compared to night; finding
   important for drug development                                                              37




                                              214
5. Diabetes

Headline                                                                                     Page #
Diabetic drug shows promise in preventing or treating diabetic kidney disease                39
Sildenafil (Viagra®) improves gastric problem as well as impotence in type-1 diabetics       41
Web technology connects diabetes patients with doctor for blood sugar monitoring             43
Levels of a hormone can predict diabetics at risk for unhealthy weight gain                  45
Large percentage of Asian Indian men in U.S. have abnormal blood sugar,
    leading to diabetes and heart disease                                                    47
First-time finding opens pathway to future diabetes treatment with American ginseng          49
Derivative of aspirin improves insulin resistance in diabetics                               51


6. Effects of Hormones on Cancer

Headline                                                                                     Page #
Testosterone added to standard HRT may reduce breast cancer                                  53
Hormones made in the heart may be future treatment for most deadly form of cancer            55


7. Gene Expression

Headline                                                                                     Page #
Specific proteins that regulate genes may improve diagnosis and treatment of breast cancer
in the future                                                                                57
Salivary glands may have role in gene therapy for inherited endocrine problems               59


8. Glucocorticoids

Headline                                                                                     Page #
Unbalanced diets in pregnant women may affect lifelong health of their children              61
Anti-inflammatory glucocoritcoids increase a person’s risk for heart disease                 63


9. Growth Hormones

Headline                                                                                     Page #
Growth hormone helps obese people lose weight and improves cholesterol levels                65
Growth hormone use in children does not increase risk of cancer                              67
Lifelong use of growth hormone does not appear to increase risk of cancer                    69
Growth hormone treatment shows lower risk of cancer and heart problems…                      71
Growth hormone test is not reliable measure in overweight and obese                          73
New drug shows improvements over existing therapy to treat life-shortening disease           75
Improved method for detecting growth hormone use by athletes may put stop to abuse           77
New family of drugs shows success in preventing atherosclerosis in early studies             79



                                              215
10. Nuclear Receptors

Headline                                                                                      Page #
New research project focuses on receptors involved in obesity, diabetes, cancer and aging     81
Discovery of the function of genetic flaw in diabetes may lead to new drug treatments         83


11. Obesity

Headline                                                                                      Page #
First-time finding shows that obese people likely have molecular abnormalities in brain       85
Hormone that controls appetite has varying effects in anorexia, bulimia and obesity           87
Low-fat dinner in young, overweight girls promotes the body’s use of fat                      89
High levels of a fat cell hormone is linked to risk of stroke in obese men                    91
Obese African Americans, especially women, have a high risk of vitamin D deficiency           93
Early research identifies potential drug targets for treating obesity and diabetes            95
Weight-loss and diabetic drugs together helped children lose weight, reduce diabetes risk     97
Combining the weight-loss power of two drugs does not add up                                  99
Newly found hormone released by stomach may serve as basis for future obesity therapy         101
Sleep restriction is linked to weight gain and risk for obesity                               103
Early study shows promise for creating weight-loss drugs based on hormone                     105
Brain peptides affect activity level and metabolism, leading to decreased weight              107
First-time finding shows fat tissue produces a hormone in obese and possibly links it to
    fat cell production                                                                       109
NeuroendocrinologyHormone administered by nose drops reduced food intake,
    caused weight loss in animals                                                             111


12. Osteoporosis

Headline                                                                                      Page #
Osteoporosis drug shows rapid and sustained ability to prevent fractures                      113
Adults who had heart transplants as children should be checked for osteoporosis               115
Menopausal estrogen loss affects risk of osteoporosis more than aging-related bone loss       117
Elderly women significantly reduce risk of bone fracture with osteoporosis drug               119
Team approach to evaluating fractures leads to better treatment for osteoporosis              121
Bone density represents small portion of osteoporosis drug’s effect on preventing fractures   123
Long-term bone strength improvements can be predicted by early markers of change
    produced by new osteoporosis drug                                                         125
Calcium deficiency in the urine identifies women at risk for bone loss and fractures          127
Vitamin D deficiency is common among adolescents, affecting lifelong bone strength            129
Too much and too little vitamin A is bad for bone strength, with increased fracture risk      131
Researchers uncover the mechanism of temporary bone loss in breastfeeding women;
    may lead to treatments for osteoporosis                                                   133




                                               216
13. Pediatric Endocrinology

Headline                                                                                       Page #
Smaller-than-expected babies, regardless of prematurity, have higher risk of adult diabetes    135
Diabetes medicine shown to help adolescents with hormone problem and prevent diabetes          137
Diabetic drug shown, for the first time, to prevent endocrine condition in young women         139
Long-term steroid treatment in children affects bone strength                                  141
Smoking During Pregnancy Increases Infants Risk of Heart Disease Later in Life                 143
Premature children have metabolic abnormalities that may lead to adult diseases                145
Children with gene alteration are more prone to obesity and precursors of diabetes             147


14. Reproduction

Headline                                                                                       Page #
Men with severe infertility likely to have genetic defects; common in vitro procedure
    may pass bad genes to offspring                                                            149
Testosterone treatment jump starts effects of Viagra in certain men                            151
Blood vessel function is impaired, leading to heart disease, in women with common
    endocrine problem                                                                          153
Testosterone spray shown to increase testosterone to healthy levels in certain women           155
Estrogen therapy improves cognitive processes in verbal learning and memory                    157
Estrogen found in Chinese healing herb, among other natural products                           159
High levels of testosterone in postmenopausal women may be culprit of increased
    heart disease risk                                                                         161
Women’s sleep is better in quantity and quality than men’s, may stem from adaptation           163
Soy infant formula may negatively affect mammary gland growth, based on animal study           165
First-time finding of mechanics of labor may lead to prevention of premature births            167
Combination of male and female hormones are viable male contraceptive                          169
Men with hormone disorder respond well to testosterone gel, regardless of testosterone
    levels at treatment                                                                        171
Sustained-released, testosterone male contraceptive appears safe, effective for future use     173
Contrary to previous research findings, male baldness is not a sign of heart disease risk      175
Sexual activity after taking sildenafil (Viagra®) may not be heart safe in some diabetic men   177
By-product of naturally occurring estrogen may help male hormone–related diseases              179
Novel protein shown to affect heart muscle growth; may hold keys to future heart therapy       181


15. Steroid Hormone Action/Biosynthesis

Headline                                                                                       Page #
New class of non-steroidal drugs may be superior treatment in common prostate problem          183
Yoga reduces levels of stress-related hormone                                                  185
New compounds have anti-inflammatory effects of steroids without side effects                  187
Weight training with male hormone therapy can improve muscle quality in
   important patient groups                                                                    189



                                               217
Headline (Steroid Hormone Action, cont.)                                                   Page #
Precedent-setting study lays groundwork for therapy of steroid-related human diseases      191
First-time finding shows that enzyme is crucial part of early development of embryo in
    mice, possibly humans                                                                  193


16. Thyroid

Headline                                                                                   Page #
Inexpensive drug for blood circulation found to prevent thyroid-related eye disease        195
Long-term intake of seaweed may contribute to thyroid problems in some                     197
Gene therapy shows promise for untreatable, aggressive form of thyroid cancer              199
Novel leukemia drug may be valuable treatment for rare, deadly form of thyroid cancer      201
Simple procedure can cure benign thyroid cysts, bypassing need for surgery                 203
Skin condition with loss of pigmentation carries high risk of autoimmune thyroid disease   205




                                              218
                   Index of Abstracts by Abstract Number


Abstract #   Author                            Page #
OR10-1       Roberto Valcavi                   203
OR20-3       Stewart G. Albert                 65
OR20-5       Patrick Wilton                    67
OR20-6       Patrick Wilton                    69
OR2-1        Shuo-Yen J. Lin                   37
OR21-1       Maurizio Bevilacqua               127
OR21-2       Catherine M. Gordon               129
OR21-3       Alexander R. Opotowsky            131
OR2-3        Carla Schulz                      111
OR23-6       Synthia H. Mellon                 193
OR28-1       Alberto Ferlin                    149
OR28-2       Antonio Aversa                    151
OR29-1       Waleed A Aldhahi                  51
OR30-2       Brian R. Walker                   63
OR32-3       Zida Wu                           77
OR32-5       Peter J. Trainer                  75
OR33-1       Plamen Penev                      103
OR33-2       Simon J.B. Aylwin                 101
OR35-4       Ping H. Wang                      27
OR36-1       Gabriella Segal-Lieberman         107
OR38-2       Antony Voutetakis                 59
OR40-1       Ronald N. Margolis                81
OR40-2       Karen M. Duda                     83
OR42-3       Jack Yanovski                     147
OR42-5       Paul L. Hofman                    145
OR49-2       Jennifer C. Condon                167
OR54-2       Harald Dobnig                     125
OR6-6        Michael Stowasser                 35
OR8-4        Rodrigo A. Bazaes                 135
P1-183       Cornelia Jaursch-Hancke           177
P1-186       Trent D. Lund                     179
P1-202       Arno W.F.T. Toorians              175
P1-258       Angelo DelParigi                  85
P1-267       David R. Powell                   95
P1-28        Britta M. Jacobsen                57


                                         219
Abstract #   Author                              Page #
P1-284       LuGuang Luo                         49
P1-345       Ricki J. Colman                     11
P1-348       Sue Rutherford                      13
P1-351       Joshua VanHouten                    133
P1-506       Nadir R. Farid                      195
P1-516       Carlos E. Fardella                  33
P1-563       Adrian H. Heald                     45
P1-570       Nicole Simon                        47
P1-597       M. Cecilia Lansang                  39
P1-609       Jutta C. Piswanger-Soelkner         41
P1-611       Robert A. Vigersky                  43
P1-82        Tony Antakly                        191
P2-153       Jorge N. Artaza                     181
P2-161       Ka Kui Lee                          169
P2-162       Andrea Coviello                     173
P2-166       Christina Wang                      171
P2-190       Evantia Diamanti-Kandarakis         153
P2-195       Hyesoo Lowe                         161
P2-208       Karen O. Klein                      159
P2-214       Hadine Joffe                        157
P2-215       Alexandros N. Vgontzas              163
P2-218       Andrew J. Humberstone               155
P2-223       Huy Ong                             79
P2-344       Johan Svensson                      71
P2-381       Tracey Bridger                      137
P2-385       Lourdes Ibanez                      139
P2-390       Outi Makitie                        141
P2-394       Peter C. Hindmarsh                  143
P2-429       Brian A. Vesely                     55
P2-446       Nadir R. Farid                      199
P2-472       M. Appetecchia                      205
P2-475       Gian Paolo Ceda                     7
P2-481       Mirjam Christ-Crain                 9
P2-487       Nelson B. Watts                     123
P2-493       Elizabeth A. Streeten               121
P2-500       A. Chines                           113
P2-501       Michael R. McClung                  119
P2-51        Bhawanjit K. Brar                   29


                                           220
Abstract #   Author                          Page #
P2-513       Jane Lukacs                     117
P2-515       Adi Cohen                       115
P2-532       Akira Ohtsuru                   201
P2-579       David I.W. Phillips             61
P2-58        Xichun Yu                       31
P3-137       Eric Hugo                       109
P3-15        Elizabeth Padilla-Banks         165
P3-196       Justin Mager                    185
P3-209       Jeffrey N. Miner                187
P3-212       Edward T. Schroeder             189
P3-221       James T. Dalton                 183
P3-424       Carolyn A. Bondy                53
P3-428       Tommy Olsson                    91
P3-434       Silvia Destefanis               87
P3-453       Claudio Maffeis                 89
P3-457       Galina Shenkerman               99
P3-458       Fuad Ziai                       97
P3-467       Shamik J. Parikh                93
P3-495       Vivien S. Bonert                73
P3-593       Selma F. Witchel                23
P3-598       Susanne Hahn                    17
P3-622       Michael Zitzmann                25
P3-628       Carolyn A. Allan                15
P3-629       Christopher J. Malkin           19
P3-632       Duarte Pignatelli               21
P3-675       Lewis E. Braverman              197
P3-93        Michael D. Culler               105
S19-1        Robert J. Anderson              1
S19-2        Eric Fliers                     3
S19-3        Manthos Giannoulis              5




                                       221
                    Index of Abstracts by Funding Source

Funding Source                                                    Page #
Abbott Laboratories                                               187
Acrux Ltd.                                                        155
American Diabetes Association                                     1, 83
American Heart Association                                        13, 23
Australian National Health and Medical Research Council           15
Australian National Heart Foundation                              15
Aventis                                                           1, 113, 119, 123
Avon Foundation, The                                              57
Biomeasure, Inc.                                                  105
Bio Technology General Corp.                                      189
Biotek                                                            173
British Heart Foundation                                          29, 63, 143
Canadian Institutes of Health Research                            37, 79, 191
Charles H. Hood Foundation                                        129
Contraceptive Research and Development Program                    169, 173
Department of Defense Research Service Command                    57
Department of Veterans Affairs Research and Development Service   1
Dunhill Medical Trust                                             61
Eli Lilly and Company                                             125
Eli Lilly Italia                                                  7
Endocrine Fellows’ Foundation                                     115
Eureka Project                                                    87
European Society for Pediatric Endocrinology                      141
Europeptides                                                      79
Fondecyt                                                          135
Fondo de Investigaciones de la Seguridad Social                   139
Foundation for Research                                           29
FSMEM                                                             87
German National Institute for Sport Science                       77
German Research Society                                           25
GlaxoSmithKline                                                   1
Government of Chile                                               33
GTx, Inc                                                          183
Hallet Diabetes Foundation                                        49
Heart and Stroke Foundation of Canada                             37



                                           222
Funding Source                                                   Page #
Hungarian Ministry of Health                                     195
Immunex/Amgen Inc.                                               31
International Olympic Committee                                  77
Italian Ministry of Health                                       7
King Faisal Specialist Hospital & Research Centre                199
King’s College Hospital Research and Development Department      101
KOS Pharmaceuticals                                              1
Lexicon Genetics                                                 95
Lifespan Development Grant                                       49
Ligand Pharmaceuticals                                           187
Lorenz Biotech                                                   127
March of Dimes                                                   193
Mayne Pharma                                                     15
McCarthy Family Foundation                                       129
Mellon Foundation                                                169
Ministerio de Sanidad y Consumo of Spain                         139
Ministero Universita e della Ricerca Scientifica e Tecnologica   151
Montreal Children’s Hospital Research Institute                  137
MURST                                                            87
National Cancer Institute                                        81
National Center on Minority Health and Health Disparities        147
National Eye Institute                                           1
National Foundation for Cancer Research                          57
National Heart Foundation of Australia                           35
National Heart, Lung and Blood Institute                         163
National Institute on Aging                                      7, 11
National Institute of Child Development                          61
National Institute of Child Health & Human Development           147
National Institute of Dental and Craniofacial Research           59
National Institute of Diabetes & Digestive & Kidney Diseases     11, 85
National Institutes of Health                                    11, 13, 23, 27,
                                                                 29, 51, 57, 83,
                                                                 103, 107, 133,
                                                                 167, 179, 181,
                                                                 189, 193
National Science Foundation                                      193
Natural Alternatives International, Inc.                         159


                                            223
Funding Source                                                          Page #
Natural Sciences and Engineering Research Council of Canada             37
Novo Nordisk                                                            1, 141
Office of Dietary Supplements of the National Institutes of Health      93
Organon                                                                 53
Osancor Biotech Inc.                                                    195, 199
P&G Pharmaceuticals                                                     113, 119, 123
Pfizer                                                                  117, 157
Pharmacia Corporation                                                   65, 67, 73, 75
Robert J. and Helen C. Kleberg Foundation                               29
Roche Diagnostic                                                        1
Scottish Executive                                                      63
Society for the Advancement of Women’s Health Research                  157
Susan G. Komen Foundation                                               197
Swedish Heart and Lung Foundation                                       91
Swedish Medical Research Council                                        71, 91
Sylvia and Charles Viertel Charitable Foundation                        35
Texas Higher Education Coordinating Board                               167
Thomas Jefferson University Center for Integrative Medicine             185
U.S. Federal Government                                                 165
Unimed-Solvay                                                           171
United Kingdom NHS North-West Research and Development Directorate      45
United States Army Medical Research and Materiel Command                43
University Hospitals of Basel, Switzerland                              9
University of Amsterdam                                                 3
University of Oklahoma Health Sciences Center                           31
University of Queensland                                                35
University of Turin, Italy                                              87
University of Verona, Italy                                             89
Veterans Affairs Medical Center Merit Review                            31
Welcome Trust                                                           101
Yale-New Haven Hospital General Clinical Research Center                161
Yale University Ethel F. Donaghue Women’s Health Investigator Program   161
Yoga Research Society, The                                              185




                                           224

								
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