13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
13th BALKAN BIOCHEMICAL BIOPHYSICAL DAYS
&
MEETING ON METABOLIC DISORDERS’
PROGRAMME & ABSTRACTS
COMMITTEES
Honorary Presidents
Engin Bermek
ġerafettin Özkurt
Executive Committee
Presiding Officials Secretariat Treasurers
Nazmi Özer AyĢe Gül Çetin Necla Öztürk
Feride Severcan Mesude ĠĢcan A.Kevser PiĢkin
Members
Hakan Aydın Nezaket Eren
Gürbüz Çelebi Nadide Kazancı
Süleyman DaĢdağ Fatma Z.Kutay
Orhan Değer Sema T.Ozan
Turk J Biochem, 2003; 28(3), 62-224 62 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
Scientific Committee
Pekcan Ungan , Coordinator Beki Kan Zehra Sayers
Ġ.Hamdi ÖğüĢ, Co-Coordinator Güldal Kırkali Vesna Svetlicic
Nurhan Puralı, Secretary Eugenia Kovacs Yusuf Tan
N.Leyla Açan Zdravko Lalchev Tijen Tanyalçın
Nejat Dalay Yahya Laleli Azmi Telefoncu
Aleksandar Dimovski Ay ÖğüĢ Aslıhan Tolun
Burak Erman H.Avni Öktem Belma Turan
Ömer Güzel Cihan Öner Engin Ulukaya
Mustafa Gültepe Ġnci Özer Figen Zihnioğlu
Vasıf N.Hasırcı Asuman Özkara Matjas Zorko
Ivan Ivanov Rüstem Nurten
Dusanka Janazic Erhan PiĢkin
Advisory Board
Kıymet Aksoy Füsun Erciyes Asuman Orçun
T.Aslan Aksu Nurten Erdal Sinan Önen
Emel Arınç Biltan Ersöz Tomris Özben
Diler Aslan Hamza Esen Meral Özgüç
Erol Atalay Sina Gökçe Ferit Pehlivan
Sevil Atasoy ġendoğan Gülen Gül Saydam
Uğur Atik Ġsmail Gülen Ahmet Sivas
Ebubekir Bakan Gül Üner Zerrin Söylemez
Oya Bayındır Münire Hacıbekiroğlu Bolkan ġimĢek
Cumhur Bilgi Levent Karaca Yavuz Taga
Bora Barutçu Baysal Karaca E.Ferhan Tezcan
Kutlay Burat Hilal Karagül Demir Tiryaki
Gülden Burçak Levent Kayrın Asuman Tokullugil
Naime Canoruç Edip Keha IĢık Türkalp
Gönenç Ciliv Nedret Kılınç Suna Türkoğlu
Salih Çelik Yüksel Koca Müjdat Uysal
Ömer Çolak ġebnem Kösebalaban Ali Yılmaz
IĢıl Çokur Mehmet Köseoğlu Piraye Yargıçoğlu
Ġlhami Demirel Yusuf KurtulmuĢ Meral Yüce
Mübeccel Durusoy Türker Kutluay Doğan Yücel
A.Kaya Emerk Sevnur Mandalcı GüneĢ Yüreğir
Ahmet Eraslan Taner Onat
Turk J Biochem, 2003; 28(3), 62-224 63 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
SCIENTIFIC PROGRAM FOR BIOCHEMICAL & BIOPHYSICAL DAYS
OCTOBER 12, 2003 SUNDAY
09:00 - 17:00 Registration
Hall C
17:00 Opening Ceremony
17:30 Opening Lecture
Chair: Nazmi Özer
Novel approaches to genetic diseases
Pınar T. Özand
18:30 Break
20:00 Welcome Cocktail
Turk J Biochem, 2003; 28(3), 62-224 64 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
OCTOBER 13, 2003 MONDAY
Hall A
Hall B
Chair: George Russev, Mesude ĠĢcan Chair: Stefan Szedlacsek, Beki Kan
Molecular Basis of Colorectal Cancer in The Republic of Exploring The Interaction of Seven-Transmembrane
Macedonia Receptors With G-Proteins By Using Synthetic Model
09:00
Alexander Dimovski Peptides
Matjaz Zorko
Irregular Ionisation of Proteins. An Alternative Oncogenic Signalling Pathways and Cell Death Based
09:40 Interpretation of Experimental Observations Tumour Therapies
Andrey Karshikoff Alexander Pintzas
From Microparticles to Giant Cells of Northern Adriatic Modelling of Cellular Receptor Signalling Pathways
10:20 Vesna Swetlicic Haluk ReĢat
Coffee Break
11:00
Chair: Vladimir Mrsa, Figen Zihnioğlu Chair: Dusanka Janezic, Kevser PiĢkin
A Simple Spectro photometric Method for the Human Interferon Gamma-Significance of the C-Terminal
11:30 determination of Selenium in Biological Materials Flexible Domain for its Biological Activity
Burcu Okutucu Genoveva Nacheva
FT-IR Spectroscopic Studies of Melatonin - Induced Non-Enzymatic Glycosylation of Recombinant Human
11:50 Changes in Rat Liver Microsomes Interferon-Gama
Güvenç Görgülü Roumyana Mironova
Molecular Investigation of The Effect of Melatonin on Rat Noninvasive Optical Assay for Synthetic Hemoglobin
12:10 Brain Tissue Fluids
Sara Banu AkkaĢ Mehmet Dinçer Bilgin
A Rat Dementia Model by Chronic Ethanol Consumption The Status of Patients with Diabetes Mellitus Monitored
and Withdrawal: Validation by Passive Avoidance by Turkish Diabetes Society in Denizli / Turkey
12:30
Measurement and Serum Cholinestrase Level Diler Aslan
Erdinç Çakır
13:00 Lunch
14:00 Poster
17:00 Free Time
Turk J Biochem, 2003; 28(3), 62-224 65 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
OCTOBER 14, 2003 TUESDAY
Hall A
Hall B
Chair: Alexander Dimovski, Zoe Lygerou Chair: Andrey Karshikoff, Gürbüz Çelebi
Global Genomic and Transcription-Coupled DNA Repair Yeast Cell Wall Proteins: Localisation, Function,
09:00 Rates in Human Cells Application
George Russev Vladimir Mrsa
Cloning, Expression and Preliminary Characterization of Catabolic and Anabolic Agents of Bone Organ - Culture
Xylulose-5-Phosphate Phospoketolase From Lactococcus Emina Sofic
09:40 lactic.
Stefan Szedlacsek
Y-Chromosome Microdeletions Among Infertile Men Electro-manipulation of The Biological Cells: Basics and
10:20 From The Republic of Macedonia Applications
Dijana Plaseska-Karanfilska Mihai Radu
11:00 Coffee Break
Chair: Ivan Ivanov, Dijana Plaseska-Karanfilska, Chair: Ivan Ivanov, Azmi Telefoncu
Ġ. Hamdi ÖğüĢ
Screening of RPGR Gene in Turkish Retinitis Pigmentosa Immunoliposomes Directed Toward VCAM-1 : Vehicles
11:30 Patients for Specific Drug Delivery to Activated Endothelial Cells
Ceren Acar Manuela Vionea
Methylation Pattern and Dnasel Hypersensitivity within the Development of two Amperometric Biosensors Based on
Intregenic Spacer of Ribosomal RNA Genes in Excised Catalase Immobilized in Gelatin - Alignate and Gelatin-k-
11:50 Cotyledons of Cucurbita Pepo L. ( Zucchini ) after Carrageenan for Alcohol Determination
Cytokinin Treatment Erol Akyılmaz
Evgeni Ananiev
Gene Polymorphism of Endothelial Markers in Renal Determination of Chromium ( VI ) by a Catalytic
Dysfunction Spectrophotometric Method in the Presence of p-
12:10
Constantina HeltianuAminobenzoic Acid
Angelina Stoyanova
Expression of Estrogen in Breast Carcinomas Altered Drug Resistance and Neurologic Disorders in
Vesna Vranic Mandusic Drosophila Melanogaster with a Deficient Histamine-Gated
12:30 Chloride Channel
Eugene Semenov
13:00 Lunch
14:00 Tour To Ephesus
Turk J Biochem, 2003; 28(3), 62-224 66 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
OCTOBER 15, 2003 WEDNESDAY
Hall A Hall B
Chair: Matjaz Zorko, Alexander Pintzas Chair: Vesna Swetlicic, Mihai Radu
The Roles of CYP1A and CYP2E in Chemical Inactivation of Melanoma Cells Irradiated with Gamma-
09:40 Carcinogenesis Rays and Low Energy Protons
Emel Arınç Alexandra Ristic-Fira
Cell Cycle Regulation and Genome Integrity Large Scale Macromolecular Simulations By Simplectic
10:20 Zoe Lygerou Integration Methods
Dusanka Janezic
11:00 Coffee Break
Chair: Alexandra Ristic-Fira, Leyla Açan Chair: Necla Öztürk, Vassilj Goltsev
The Effect of Insulin Level on Steroid Receptors Implementation of a Neovascular Coupling Model
11:30
Mojka Vulovic Ata Akın
Molecular Analysis of the ABCA4 Gene in Turkish Analysis of Millisecond Delayed Fluorerescene Dark
11:50 Patients with Stargardt Disease and Retinitis Pigmentosa Decay
Rıza Köksal Özgül Vassilj Goltsev
Mitochondrial Enzyme Activities and mtDNA Development of an Analogue Model Simulating the Portal
12:10 Polymorphism in Parkinson's Disease Vein
Gülnihal Kulaksız Necla Öztürk
Determination of Steadt-State Levels of OxoGuanine in Purification and Characterization of A PSTS Hyperalkaline
12:30 Calf Thymus DNA by Means of FPG Protein Phosphates from T.Thermophilus
Beran YokuĢ Anastasia Pantazaki
13:00 Lunch
14:00 Poster
Closing Ceremony
17:30 Free Time
20:00 Farewell Dinner
Turk J Biochem, 2003; 28(3), 62-224 67 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
SCIENTIFIC PROGRAM FOR MEETING ON METABOLIC DISORDERS
OCTOBER 13 , 2003 MONDAY
Hall C
Chairmen: A. Kaya Emerk, IĢık Türkalp
Ortadoğuda Genel Problem Olan Homosistinürinin üç Tipi
09:00 Three Types of Homocystinuria A Common Problem in Middle East
Pınar T. Özand
Yenidoğanda Konjenital Hipotiroidi Taraması
10:05 Screening for Congenital Hypothyroidism in theNeonate
Alev Özön
Yenidoğanda Tandem-Mass Spektrometre ile GeniĢ Kapsamlı Kalıtsal Metabolik Hastalıkların Taranması
10:20 Expanded Newborn Screening for inborn Errors of Metabolism by Tandem Mass Spectrometry
Ġnci Karaaslan
10:40 TartıĢma/ Discussion
11:00 Coffee Break
Chairmen: Fatma Z. Kutay, Nezih Hekim
Kitle Tarama Testlerinin UygulanıĢında Kalite Kontrolunun Önemi ve Kitle Tarama Verilerinde Referans
Aralığının Saptanması
11:30
The Use of Split-Sample Design for Performance Evaluation of Screening Kits: A Real Life Study
Tijen Tanyalçın
Fetusta Biyokimyasal Tanı
11:50 Foetal Biochemistry: Biochemical Diagnosis for Foetus
Nezih Hekim
12:10 TartıĢma / Discussion
13:00 Lunch
14:00 - 16:30 Poster
Panel : Türkiye'de Neonatal Tarama Politikası
Neonatal Screening Policy of Turkey
17:00 - 19:00
Chair: Uğur Dündar / Mine Özbek
Pınar T. Özand, Ġmran Özalp, Münip Üstündağ, Yahya Laleli, Mine Özbek
Turk J Biochem, 2003; 28(3), 62-224 68 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
OCTOBER 14 , 2003 TUESDAY
Hall C
Chairmen: Yahya Laleli, Tijen Tanyalçın
Amino Asit Metabolizması Bozukluğunda Kullanılan Metodlar ve KarĢılaĢılan Güçlükler
09:45 Amino Acid Analysis Methods-Details and Difficulties
Gürsel Biberoğlu
The Use of Tandem Mass Spectrometry in Newborn Mass Screening Programs: The Facts Beyond the Myth
10:15
François Eyskens
11:00 Coffee Break
Chairmen: Aslan Aksu, Sema T. Ozan
Karbohidrat Metabolizması Hastalıkları: Klinik YaklaĢım, TeĢhis ve Tedavide Laboratuvardan Beklentiler
Disorders of Carbohydrate Metabolism : Clinical Approach
11:30
Future Prospects of Diagnosis and Treatment
Benal Büyükgebiz
Karbohidrat Metabolizması Hastalıkları: Metodlar ve KarĢılaĢılan Güçlükler
Disorders of Carbohydrate Metabolism Basic Concepts, Evaluation of Laboratory Methods and Difficulties
12:00
Observed
Aslan Aksu
12:30 TartıĢma / Discussion
13:00 Lunch
14:00 Tour to Ephesus
Turk J Biochem, 2003; 28(3), 62-224 69 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
OCTOBER 15, 2003 WEDNESDAY
Hall C
Chairmen: Güldal Kırkalı, Hakan Aydın
Obesite, Polikistik Over ve Ġnsülin Direnci
09:00 Dsyregulation of P450C17 Enzyme in Polycystic Ovary Syndrome
Fahrettin KeleĢtimur
Obesite: Nöral Mekanizmalar
09:30 Neuroendocrine System and Obesity
Üstün Korugan
Obesite Bir Ġnflamasyon mudur?
10:00 Is Obesity an Inflammatory Disease ?
Candeğer Yılmaz
10:30 TartıĢma / Discussion
11:00 Coffee Break
Chairmen: Orhan Değer, Nezaket Eren
Hiperlipoproteinemiler ve Diğer Lipid Metabolizması Hastalıklar
11:30 Laboratory Diagnosis in Lipoprotein and Other Lipid Metabolism Disorders
Gülçin Eskandari
Glikosfingolipidozlar: Hastalıktan Metabolizmanın Temel Ġlkelerine
12:00 The Glycosphingolipidoses: From Disease to Basic Principles of Metabolism
Asuman Özkara
12:30 TartıĢma / Discussion
13:00 Lunch
14:00 Poster
17:00 Closing
17:30 Free Time
20:00 Fairwell Party
Turk J Biochem, 2003; 28(3), 62-224 70 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
2) Institute for Pharmaceutical Chemistry, Faculty of
OCTOBER 12, 2003 - SUNDAY Pharmacy, Skopje, Republic of Macedonia
HALL C Correspondence: adimovski@baba.ff.ukim.edu.mk
OPENING LECTURE Colorectal cancer (CRC) is one of the most common
cancers and the second cause of death in developed
NOVEL APPROACHES TO GENETIC countries. In addition to environmental factors, genetic
DISEASES predisposition has a significant role in the
Pinar T. Ozand MD,Ph. D., Chairman, ethiopathogenesis of the disease. Apart from the two
dominantly inherited syndromes (FAP and HNPCC)
Department of Genetics, King Faisa Specialist
several low penetrance genes were implicated in the
Hospital & Research Centre. PO Box 3354, Riyadh 11211,
initiation of colorectal cancerogenesis. The aim of this
Saudi Arabia.
study was to determine the molecular basis of FAP, the
The objective of this presentation is to present the incidence of HNPCC and the frequency of polymorphisms
novel strategies adopted to prevent genetic diseases in in several low penetrance genes (I1307K and E1317Q in
Saudi Arabia. The genetic diseases either due to the APC gene, TRI(6A) and CCND1) associated with
chromosome defects or to alterations in single gene CRC. A total of 173 patients with CRC, of which six
structure produce infants with usually severe diseases. patients with multiple adenomatous polyposis, and a
The management of such disorders is: (A) usually costly control group of 100 newborns and 100 aged individuals
due to the fact the morbidity lasts for many years, (B) not were included in this study. Out data indicate that FAP and
always successful and doesn‘t always produce good HNPCC have relatively low frequency of 0.1% and 5x106/ml). The Evgueni D. ANANIEV1, Galina ABDULOVA1, Petar
prevalence of Y microdeletions among the infertile males GROZDANOV2, Luchezar KARAGYOZOV2
from the Republic of Macedonia is 6.4%, among patients 1
Institute of Plant Physiology “M. Popov”, 2 Institute of
with azoospermia – 17.4% and among those with severe Molecular Biology, Bulg. Acad. Sci., “Acad. G. Bonchev”
oligozoospermia - 3.6%. Different testicular defects were str., block 21, 1113, Sofia/ BULGARIA
found among the patients with AZFc deletions (SCOS, ananiev@obzor.bio21.bas.bg
hypospermatogenesis and maturity arrest). Testicular
―Run-on‖ transcription experiments with isolated nuclei
volumes were reduced in the patients with Y
have showed that treatment of detached marrow cotyledons
microdeletions, FSH concentrations were high and LH and with the cytokinin 6-benzyladenine (BA) resulted in 2-4-
testosterone were within the normal range.
Turk J Biochem, 2003; 28(3), 62-224 75 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
fold stimulation of RNA polymerase I activity. In this work followed by agarose gel electrophoresis. The Glu298Asp
we studied the methylation pattern of the intergenic spacer of eNOS was assessed by RFLP-PCR technique and then
(IGS) of rRNA genes as a measure of their activity using separated by polyacrilamide gel electrophoresis.
the restriction enzymes Msp I and Hpa II and the method Results: There is no association of eNOS and ACE gene
of ―indirect end labeling‖. A cloned fragment containing variants with the presence of renal dysfunction in diabetic
the 5/ portion of 18S rRNA gene from flax was used as patients. In the case of Fabry disease, both eNOS gene
DNA probe. Results showed heavy methylation of the polymorphism were significantly associated with the
rRNA genes. As judged from the almost total lack of affliction namely P=0.02, OR=2.7, 95%CI=1.1-6.4 for
digestion with Hpa II, in the repeating rDNA units there Glu298Asp, and P=0.05, OR=2.6, 95%CI=1.0-7.0 for
were either no methylation free regions or just a few were intron 4 VNTR. Carriers of eNOS 298Asp allele were
observed. A hypomethylated Hpa II site near the promoter overrepresented (P=0.04) in Fabry subgroup with renal
region was detected in a very small number of rDNA failure when compared with patients without it, giving a
repeats. Digestion with Msp I affected nearly 50% of the 4.64-fold increased risk (95% CI=2.43-6.85) of the
repeating units. This suggested that in addition to –CpG- incidence of renal dysfunction. The distribution of ACE D
sequences, methylation in –CpNpG- might not be random. allele was similar among the Fabry patients and control
The methylation pattern of IGS was not changed upon subjects.
hormonal treatment of cotyledons DNase I assay with
isolated nuclei revealed defined regions of DNase I Conclusion: These findings suggest that the eNOS
hypersensitivity in IGS which coincide with the regulatory polymorphism might influence the development of renal
elements. The DNase I hypersensitive sites in IGS were failure in Fabry disease.
mapped to the active promoter (transcription initiation site This project was financially supported by the Romanian
–TIS), to transcription termination site (TTS) and the Academy (GAR 125) and INSERM France (PECO 637).
―spacer‖ promoter region. No appreciable differences in
DNase I hypersensitivity of IGS were found after hormone ORAL PRESENTATION 4
treatment of cotyledons nor in differentiated leaves of
intact seedlings. The obtained results are discussed with EXPRESSION OF ESTROGEN RECEPTOR β
respect to the possible molecular mechanisms of cytokinin IN BREAST CARCINOMAS
regulation of rRNA gene transcription.
Vesna VRANIģ MANDUSIģ1, Dusan POPOV-
CELKETIģ1, Ksenija KANJER2 and Dragica NIKOLIģ-
ORAL PRESENTATION 3
VUKOSAVLJEVIģ2
GENE POLYMORPHISM OF 1 ”Vinča” Institute of Nuclear Sciences, Laboratory for
ENDOTHELIAL MARKERS IN RENAL Radiobiology and Molecular Genetics, Belgrade, Serbia
DYSFUNCTION and Montenegro
Constantina HELTIANU, Gabriela COSTACHE, Mihaela 2 Institute for Oncology and Radiology of Serbia,
BODEANU, Alexandra DOBRIN, Kemal AZIBI*, Livia Belgrade, Serbia and Montenegro
POENARU*, Maya SIMIONESCU
vvranic@vin.bg.ac.yu
Institute of Cellular Biology and Pathology
“N.Simionescu”, Bucharest, ROMANIA, *Department of To determine whether estrogen receptor beta (ERβ)
Genetic, Faculty of Medicine Cochin Port-Royal, expression is associated with estrogen receptor α (ERα)
University Rene Descartes, Paris, FRANCE and progesterone receptor (PR) status we compared its
iheltianu@simionescu.instcellbiopath.ro expression with levels of ERα and PR. Thirty two samples
of breast carcinomas were examined for ERα, ERβ and PR.
Introduction: Renin-angiotensin system is implicated in Levels of ERα and PR were measured by conventional
the progression of kidney disease in diabetes mellitus
biochemical techniques. Expression of ERβ mRNA was
(DM). Another disease in which the kidneys are
determined by RT-PCR. Briefly, RNA from tumor samples
dramatically affected is the monogenic X-linked Fabry
was isolated according the method of Chomczynski and
disease. The Latter is characterized by progressive
accumulation of lipids (due to the deficiency of alpha- Sacchi (1987); total RNA was reverse transcribed with
galactosidase A activity) preferentially in vascular oligo dT primer; PCR was performed using the specific
endothelial cells (EC) which become activated. primers positioned in exons 4 and 6. PCR (254 bp product)
were analyzed on polyacrilamide gel electrophoresis. In 9
Aim: The purpose of the study was to analyze whether the carcinomas ERβ mRNA was not detected, and 23
gene encoding endothelial markers such as endothelial carcinomas (71,8%) were positive for ERβ expression.
nitric oxide synthase (eNOS), and angiotensin converting
Level of expression was quantified densitometricaly and
enzyme (ACE) influence the development of nephropathy
relatively ranged from 1 to 3. Results conform positive
in DM and Fabry disease.
correlation between ERα and PR status. Analysis of
Methods: To test this hypothesis about 200 DM subjects, correlation between ERα and ERβ expression as wel as
28 hemizygot unrelated Fabry patients and 120 healthy ERβ expression and PR shows:
individuals were genotyped for the intron 4VNTR and
Glu298Asp of eNOS, and I/D variant of ACE to analyze absence of any correlation between ERα and ERβ;
their association with the disease. The 4VNTR eNOS and statistical significant (hi-square test, p=0,03) inverse
I/D ACE variants were determined by PCR amplification correlation between ERβ and PR status.
Turk J Biochem, 2003; 28(3), 62-224 76 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
Therefore, expression of ERα and ERβ are independent the predominant human CYP enzyme involved in the
one from the other, but increased expression of ERβ is activation of AFB1. In the presence of hepatitis B virus
connected with decreasing of ER functionalities (HBV), a relative risk for liver cancer upon exposure of
represented through decreased PR expression. AFB1 increased 30-fold.
CYP2E1 is also involved in chemical carcinogenesis, both
OCTOBER 15 , 2003 - WEDNESDAY in the metabolic activation of small molecular weight
organic chemicals and in the generation of ROS. N-
HALL A Nitrosodimethylamine (NDMA) is a procarcinogen that is
LECTURE 1 activated by CYP2E1 dependent NDMA N-demethylase.
NDMA is known to be carcinogenic in liver, kidney and
THE ROLES OF CYP1A AND CYP2E IN lung. Pyridine, constituent of tobacco and tobacco smoke,
CHEMICAL CARCINOGENESIS has tumor-promoting and teratogenic activities. Following
Emel ARINÇ in vitro pyridine treatment of rabbits, NDMA N-
demethylase activity of both liver and lung is stimulated by
Graduate Program in Biochemistry, Department of 6.9- and 5.2-fold, respectively indicating that pyridine
Biology, Middle East Technical University, 06531, Ankara, increases the metabolic activation of NDMA and in turn,
TURKEY may potentiate formation of liver and lung cancers
earinc@metu.edu.tr significantly (Arınç, E., Adalı, O., Gençker-Özkan, A. M.:
Arch. Toxicol., 34, 329-334, 2000). In addition,
It has been established that approximately 80% of human
experiments carried out in our laboratory demonstrated that
cancers are attributable to environmental agents and about
in the experimentally induced diabetic rabbits, both
70% of the human cancers are caused by chemical
CYP2E1 and NDMA N-demethylase activity have been
compounds. The majority of the organic carcinogens is not
increased about two-fold in kidney and liver of rabbits.
reactive as such but must undergo enzymatic reactions to
Therefore, it is expected that the incidence of tumor
form electrophilic species. The first step in
formation due to exposure to NDMA will be more
biotransformation is usually the oxidative step, catalyzed
pronounced in diabetic state because of CYP2E1 induction.
by microsomal cytochrome P450 (CYP) dependent
monooxygenases. Upto now more than 2000 P450 genes
are sequenced and 265 P450 families have been identified LECTURE 2
and 18 of them are found in humans. Although other P450s CELL CYCLE REGULATION AND
and other mechanisms may activate the carcinogens, GENOME INTEGRITY
CYP1A enzymes are the most significant and 90% of the
known carcinogens are metabolically activated by CYP1A Z. Lygerou
to the proximate and ultimate carcinogens. Department of Gen Biology, Medical School, University of
Persistent organic pollutions such as polyaromatic Patras, Greece
hydrocarbons (PAHs), dioxins, polychlorinated biphenyls lygerou@med.upatras.gr
(PCBs) specifically induce CYP1A1 through Ah receptor
For cells to maintain genome integrity, S-phase must
mediated mechanism. At the same time, CYP1A1 mostly
alternate with mitosis at every cell cycle. Licensing of
converts PAH- and PCB-type precarcinogens to their
DNA for replication takes place only after completion of
carcinogenic epoxides or other oxygenated metabolites.
mitosis and ensures once per cell cycle replication.
These epoxides, in turn bind DNA covalently forming
DNA-adducts which may cause cancer in the years to Cdt1, a central licensing factor conserved across evolution,
come. Thus induction of CYP1A1 is used to measure both is tightly controlled in human cells, so that it is present
exposure and resulting toxic carcinogenic effects of these only in the G1 phase, when licensing is legitimate. Over-
types of organic pollutants. (Arınç, E., ġen, A., expression of Cdt1 leads to genomic instability. We have
Bozcaarmutlu, A.: Pure & Appl. Chem. 72, 985-994, shown that ubiquitin dependent proteolysis ensures that
2000). Greater CYP1A1 induction may result in high levels Cdt1 is destroyed as soon as S-phase starts. Cdt1 is
of activated carcinogens, and consequently to higher undetectable in cells accumulating cyclin A, suggesting
degree of persistent DNA-adduct formation or to enhanced that phosphorylation by cdk2/cdc2 – cyclin A complexes
oxidative DNA damage. targets Cdt1 for degradation. Geminin, a molecular
inhibitor of Cdt1, accumulates in cells which do not
Good correlation between smoking and lung cancer is
contain Cdt1, suggesting that Geminin‘s inhibitory
established. Individuals with a high inducible phenotype
function over Cdt1 might be redundant for the majority of
CYP1A1 show high risk of lung cancer. Gene-environment
the mammalian cell cycle. When primary cells exit the cell
interactions are more pronounced at lower levels of
cycle to G0, both Cdt1 and Geminin protein and mRNA
cigarette exposure in which the susceptibility to lung
levels are decreased. In contrast, cancer cells over-express
cancer increased in the case of patients with either
both these proteins.
CYP1A1 mutated alleles and GSTM1 null gene.
In order to study the localization, dynamics and
On the other hand, CYP1A2 activates many arylamines
interactions of licensing factors in the living cell, we
and amides present in food and cigarette smoking.
constructed fusions of Cdt1 and Geminin to Green
CYP1A2 exhibits polymorphisms and inducibility.
Fluorescent Protein (GFP), Cyan Fluorescent Protein
Aflatoxin B1 (AFB1) must be activated to a reactive
(CFP) and Yellow Fluorescent Protein (YFP) and
epoxide prior to exerting carcinogenic effects. CYP1A2 is
characterized their behavior after transfection into
Turk J Biochem, 2003; 28(3), 62-224 77 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
mammalian cultured cells. Time-lapse microscopy and ORAL PRESENTATION 2
fluorescence lifetime imaging microscopy (FLIM) allows
us to identify the location and timing of protein-protein and MOLECULAR ANALYSIS OF THE ABCA4
protein-DNA interactions as cells progress through the cell GENE IN TURKISH PATIENTS WITH
cycle. STARGARDT DISEASE AND RETINITIS
PIGMENTOSA
OCTOBER 15, 2003 – WEDNESDAY Rıza Köksal ÖZGÜL1, Hakan DURUKAN2, AyĢe
TURAN3, Cihan ÖNER1, Ay ÖĞÜġ1, Debora B.
HALL A FARBER4
ORAL PRESENTATION 1 1 Hacettepe University, Department of Molecular Biology,
Beytepe-Ankara,Turkey
THE EFFECT OF INSULIN LEVEL ON Gülhane Military Medical Academy, Department of
STEROID RECEPTORS Ophthalmology, Ankara,Turkey
Mojca VULOVĠģ, Goran KORIģANAC, Sneţana Numune Research and Training Hospital, Department of
RADIVOJŠA‚ Zorica ŢAKULA, Nevena RIBARAC- Ophthalmology, Ankara,Turkey
STEPIģ University of California, Los Angeles (UCLA), Jules Stein
Laboratory for Molecular Biology and Endocrinology, Eye Institute , Los Angeles, USA
“Vinča“ Institute of Nuclear Sciences, P.O. Box 522, rkozgul@hacettepe.edu.tr
11001 Belgrade/SERBĠA AND MONTENEGRO Stargardt disease (STGD), the most prevalent autosomal
mojca@rt270.vin.bg.ac.yu recessively inherited macular dystrophy is characterized by
severe reduction of central visual acuity and leading to
Steroid receptors are key molecules in steroid hormone partial or complete blindness due to photoreceptor
action. It has been proposed that a nonsteroidal hormone degeneration. The responsible gene for STGD encodes a
such as insulin may exert an influence on steroid receptors ATP-binding cassette transporter, ABCA4 which has been
and alter the cell responses to steroid hormone action. shown to be involved in retinoid transport in the retina.
Diabetes mellitus is very frequent metabolic disorder that Molecular characterization of the ABCA4 gene led to the
also has implications on steroid hormone action. The main identification of many mutations in Stargardt disease,
goal of the presented study was to elucidate the effects of cone-rod dystrophy (CRD), autosomal recessive retinitis
insulin treatment and insulin deficiency on steroid pigmentosa (arRP) and age- related macular degeneration
receptors. (AMD). By the screening of the ABCA4 gene in various
The experiments were performed on 3-months-old male types of inherited retinal degenerations, more than two
Wistar rats. Intact and streptozotocin-pretreated rats hundred disease-associated mutations reported in different
(60mg/kg, 7 days before hormone treatment) were injected populations.
In this study, all fifty exons and flanking intronic
sacrifice. Protein contents of estrogen (ER), progesterone sequences of the ABCA4 gene were screened by Single
(PR), androgen (AR) and glucocorticoid receptors (GR) in Strand Conformation Polymorphism (SSCP) in a cohort of
liver were analyzed by immunoblotting method using 40 Turkish patients with STGD and arRP. After SSCP
appropriate specific antibodies. analysis, DNA fragments showing different migration
patterns were sequenced.
The obtained results point out that streptozotocin did not
cause significant changes in protein content of any kind of Our results revealed the presence of three novel mutations
analyzed steroid receptors. Insulin injection significantly (C54G, T829M, IVS19-6C>A), two mutations previously
decreased the AR protein content in intact rats, while there reported (R212C, IVS28+4C>T) and several polymorphic
were no changes in protein content of other three analyzed changes in the ABCA4 gene among Turkish patients
steroid receptors in rat liver. Insulin given to insulin- affected with Stargardt and arRP.
deficient animals did not cause changes in steroid receptor This is the first report on the mutation profile of the
content compare to intact rats. However, in comparison to ABCA4 gene in Turkish patients. Further studies will be
diabetic rats there was the significant decrease in PR helpful in understanding of complex genotype- phenotype
content, whereas protein contents of other steroid receptors relationship in ABCA4 gene in our population.
were unchanged.
ORAL PRESENTATION 3
The presented data indicate that insulin treatment and
insulin deficient state did not alter the protein content of MITOCHONDRIAL ENZYME ACTIVITIES
ER and GR in rat liver. The levels of PR and AR were not AND MtDNA POLYMORPHISMS IN
influenced by streptozotocin treatment, while, depend on PARKINSON’S DISEASE
endogenous level of insulin, hormone treatment decreased
their content. Meltem MÜFTÜOĞLU1, Özlem DALMIZRAK1, Bülent
ELĠBOL2, AyĢe ERCAN1, Gülnihal KULAKSIZ1, Hamdi
The research was supported by grant from Ministry of ÖĞÜġ1, Turgay DALKARA2 and Nazmi ÖZER1
Science, Technology and Development, Republic of
Serbia: ‖Insulin and steroid receptors as mediators of Hacettepe University, Faculty of Medicine, 1Department of
hormone actions and their cross-talk under physiological Biochemistry,
and nonphysiological conditions‖. (Grant number 1999). 2
Department of Neurology, 06100 Sihhiye, Ankara, Turkey.
Turk J Biochem, 2003; 28(3), 62-224 78 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
Parkinson‘s disease (PD) is a progressive formamidopyrimidine-DNA glycosylase) is a combined
neurodegenerative movement disorder characterized DNA glycosylase-AP lyase that removes the damaged
clinically by tremors, rigidity, postural instability and bases (fapy-pyrimidine and 8-OxoGua lesions). The
bradykinesia. Idiopathic PD is the most common form of oxidized DNA base 8-OxoGua has been commonly
parkinsonism, but its etiology is still unknown. measured by enzymatic hydrolysis of DNA followed by
Mitochondrial dysfunction that generates oxidative stress reverse phase HPLC-EC. There has been recently a debate
contributes to the etiology of idiopathic PD. The reduced surrounding the validity of this approach, from which it has
complex I activity, that is one of the basic abnormalities become clear that artifactual oxidation of the native base to
responsible for mitochondrial dysfunction, has been 8-OxoGua that can occur at numerous stages in sample
reported in PD not only in the substantia nigra but also in preparation.
peripheral tissues. However, there are contraversing studies Hence, we developed an alternative/modified method to
about the decrease in complex I activity in peripheral traditional enzymatic digestion of DNA, which based on
tissues of idiopathic PD, since methodological factors such the use of the base excision repair enzymes (Fpg protein)
as the use of homogenates or purified mitochondria or age and limits the potential for artifactual oxidation and speeds
differences between patients and control group affect those up the assay. In addition, we showed that substrate
enzyme activities. In order to eliminate these factors, in specifity of fpg protein.
this study, we purified mitochondria from leukocytes of
idiopathic PD patients and analyzed mitochondrial All chemicals purchased from Sigma. Calf Thymus DNA
complex I and complex IV enzyme activities in these was dissolved in 20 mM TE buffer (pH 7,4),. Different
purified mitochondrial suspensions to evaluate the concentrations of the calf thymus DNA was incubated with
functional activity of the mitochondrial respiratory 16 μl Fpg protein 37° C for 2 h and hydrolysate was
enzymes. In addition, ND2 subunit of complex I enzyme analysed by HPLC for 8-OxoGua using electrochemical
were analyzed to identify 5460G/A polymorphism in both detection (Decade, Antec-Leyden). Guanine was detected
leukocytes and platelets of thirthyseven Turkish idiopathic with UV/Visible spectrophotometric (Shimadzu) detector.
PD patients and 100 healthy subjects. We found a Results were given as 8OxoGua / Gua. Retention time of 8-
statistically significant decrease in complex I (55 %) and OxoGua was 4,8. Km value 7 nm as calculated from the
complex IV (58 %) enzyme activities in the leukocytes of Lineweaver-Burk plot. In conclusion, excision enzymes
idiopathic PD patients. But, there was no significant have proved useful tools for the determination of the yield.
correlation between these enzyme activities and age, age of
onset and duration of the disease. The frequency of
5460G/A polymorphism was found to be 0.08 (3/37) in the OCTOBER 13, 2003 – MONDAY
idiopathic PD patients and 0.10 (10/100) in the control HALL B
group. Thus, no effect of ND2 G5460A genotype on
complex I enzyme activity was detected. The observed LECTURE 1
respiratory chain enzyme deficiency supports the EXPLORING THE INTERACTION OF
hypothesis that systemic mitochondrial dysfunction is
SEVEN-TRANSMEMBRANE RECEPTORS
important in the pathogenesis of idiopathic PD.
WITH G-PROTEINS BY USING SYNTHETIC
ORAL PRESENTATION 4 MODEL PEPTIDES
Matjaţ ZORKO
DETERMINATION OF STEADY-STATE
LEVELS OF 8-OxoGuanine IN CALF Medical Faculty, Institute of Biochemistry, 1000 Ljubljana,
Vrazov trg 2, Slovenia
THYMUS DNA BY MEANS OF FPG
PROTEIN zorko@mf.uni-lj.si
Beran YOKUġ1, Naime CANORUÇ2, Dilek Ülker Synthetic model peptides derived from the intracellular
ÇAKIR2, YILDIZ ATAMER2, Abdurrahman KAPLAN2, parts of different seven-transmembrane receptors were
Sabri BATUN2 used to assign the regions of the receptors that interact with
1 G-proteins.
Dicle University, Veterinary Faculty Department of
Biochemistry, Diyarbakir To determine the domains essential for G-protein coupling
2 of the human galanin receptor type 1 (GalR1), we used
Dicle University, Medical Faculty Departments of GalR1 mutants and synthetic receptor-derived peptides in
Biochemistry, Diyarbakir 125
I-galanin and [35S]-GTPS binding studies. Peptides
7,8-dihydro-8-oxoguanine (8-Oxoguanine or 8-OxoGua), a derived from the third intracellular loop (IC3), especially
major DNA damage resulting from oxidative attack, is its N-terminal part, increased the rate of [35S]-GTPS
highly mutagenic leading to translation of GC→AT . DNA binding to the trimeric Gi1, but not to Gs, Go, and G11;
adduct are lethal if not repaired. The primary function of peptides corresponding to the first and the second
Base excision repair (BER) enzymes are known to intracellular loops (IC1 and IC2) had no such effect. IC3
recognise various types of base damage: oxidised purine, also inhibited the binding of 125I-galanin to GalR1. This
pyrimidine damages and remove these ox datively suggests that the N-terminal part of IC3 defines the
damaged bases from DNA, protecting cells from the coupling of GalR1 to Gi1 and consequently, to the signal
mutagenic and lethal effects of oxidative DNA damage. transduction cascade.
Escherichia coli Fpg protein (also known as
Turk J Biochem, 2003; 28(3), 62-224 79 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
Peptides corresponding to IC1, IC2, and IC3 of glucagon- Psichari, E., Balmain, A., Plows, D., Zoumpourlis, V., and
like peptide-1 receptor (GLP-1R) showed differential Pintzas, A. (2002). High activity of serum response factor
effects on various G proteins. Results suggest much more in the mesenchymal transition of epithelial tumor cells is
complex coupling of GLP-1R to G-proteins in comparisson regulated by RhoA signaling. J. Biol. Chem. 277, 29490-
to GalR1. GLP-1R is coupled to Gs, Gi1, Go, and G11. 29495.
IC3 is the main switch that mediates signalling via GLP-1R
4. Zoumpourlis, V., Papassava, P., Linardopoulos, S.,
to G-proteins, while IC1 and IC2 are important in
Gillespie, D., Balmain, A., and Pintzas, A. (2000). High
discrimination between different types of G-proteins. We
levels of phosphorylated c-Jun, Fra-1, Fra-2 and ATF-2
have found a new potential level of GLP-1R modulation by
the endogenous ADP-ribosylase, since IC3 peptide is a proteins correlate with malignant phenotypes in the
good substrate of this enzyme and it also competes with the multistage mouse skin carcinogenesis model. Oncogene
pertusis toxin sensitive G-proteins for ADP-rybosylation. 19, 4011-4021.
Knowledge from the presented and other studies was used
LECTURE 3
to design the receptor derived synthetic peptides with the
potential to regulate some physiological processes, as MODELING OF CELLULAR RECEPTOR
demonstrated by ex vivo functional studies on the isolated SIGNALING PATHWAYS
tissues.
Haluk RESAT and H. Steven WILEY
LECTURE 2 Biological Sciences Division, Pacific Northwest National
Laboratory, Richland, WA 99352 USA
ONCOGENIC SIGNALLING PATHWAYS
haluk.resat@pnl.gov
AND CELL DEATH BASED TUMOUR
THERAPIES Endocytic trafficking of many types of receptors can have
profound effects on subsequent signaling events.
Roberts M. , Moumtzi, S. , Drosopoulos, C., Psahoulia, F., Quantitative models of these processes, however, have
Psarras, I., Papadogiannakis, I. and Pintzas, A. E-mail: usually considered trafficking and signaling independently.
apint@eie.gr Here, we present an integrated model of both the
Laboratory of Signal Mediated Gene Expression, Institute trafficking and signaling pathway of the epidermal growth
of Biological Research and Biotechnology, National factor receptor (EGFR) using a probability weighted-
Hellenic Research Foundation, 48, Vas. Constantinou dynamic Monte Carlo simulation. Our model consists of
Avenue, 116 35 Athens, Greece. hundreds of distinct endocytic compartments and about
Constitutively active forms of Ras are found in a variety of 13,000 reactions/events that occur over a broad spatio-
tumours (1) suggesting an important role for this pathway temporal range. By using a realistic multi compartment
in cancer. Ras activates the MEK-ERK pathway and model, we can investigate the distribution of the receptors
activated ERK1/2 translocate to the nucleus where they among cellular compartments as well as their potential
phosphorylate a variety of targets . signal transduction characteristics. Our new model also
allows the incorporation of physio-chemical aspects of
We have developed conditionally mouse and human cell
systems of activated V12 mutant Harvey Ras oncogene ligand-receptor interactions, such as pH-dependent binding
expression. Here we report for the first time that in the in different endosomal compartments. To determine the
absence of growth factors initial cellular exposure to utility of this approach, we simulated the differential
oncogenic Ras only transiently activated the same pathway activation of the EGFR by two of its ligands, epidermal
in the nucleus by a mechanism which involves the growth factor (EGF) and transforming growth factor- alpha
phosphotyrosine phosphatase MKP-1 (2). We have also (TGF-a). Our simulations predict that when EGFR is
investigated the impact of transient nuclear MAPK activated with TGF-a, receptor activation is biased toward
activation on the cell cycle as well as to changes in global the cell surface whereas EGF produces a signaling bias
gene expression profiles by using high density (microarray) towards the endosomal compartment. Experiments confirm
analysis. We have also compared early events in Ras these predictions from our model and simulations. Our
signalling with late nuclear effects of Ras associated with model accurately predicts the kinetics and extent of
cell transformation (3, 4). The interplay between
receptor down-regulation induced by either EGF or TGF-a.
proliferative and apoptotic signals mediated by Ras are
Our results suggest that receptor trafficking controls the
going to be discussed.
compartmental bias of signal transduction, rather than
1. Bos, J. L. (1989) Ras oncogenes in human cancer: a simply modulating signal magnitude. Our model provides a
review. Cancer Res. 49, 4682-4689. new approach to evaluating the complex effect of receptor
Plows, D., Briassouli, P., Owen, C., Zoumpourlis, V., trafficking on signal transduction. Importantly, the
Garrett, M. and Pintzas, A. (2002). Conditional activation stochastic and compartmental nature of the simulation
of oncogenic Ras leads to transient nuclear localisation of allows these models to be directly tested by high-
activated ERK regulated by MKP-1. Biochem. J. 362, 305- throughput approaches, such as quantitative image
315. analysis.
Turk J Biochem, 2003; 28(3), 62-224 80 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
OCTOBER 13, 2003 – MONDAY takes place in Escherichia coli as well. In the present study
we demonstrate that the post-translational processing
(proteolysis and covalent dimerization) observed with the
ORAL PRESENTATION 1 cysteineless recombinant human interferon-gamma (rhIFN-
) is tightly associated with its in vivo glycation. Our
HUMAN INTERFERON GAMMA: results showed that at the time of isolation rhIFN-
SIGNIFICANCE OF THE C-TERMINAL contained early but not advanced glycation end products
FLEXIBLE DOMAIN FOR ITS BIOLOGICAL (AGEs). Using RP-HPLC in conjunction with fluorescence
ACTIVITY measurements, ELISA and mass spectrometry we found
Genoveva NACHEVA1, Kristina TODOROVA1, Maya that AGEs arose in rhIFN- on storage. The latter were
BOYANOVA1, Alfredo BERZAL-HERRANZ2, Andrey identified mainly in the Arg/Lys rich C-terminus of the
KARSHIKOFF3 and Ivan IVANOV1 protein, which was also the main target of proteolysis.
1 Mass spectral analysis and N-terminal sequencing revealed
Institute of Molecular Biology, Bulgarian Academy of four major (Arg140/Arg141, Phe137/Arg138,
Sciences, Acad. G. Bonchev Str., 21, 1113 Sofia, Bulgaria; 2 Met135/Leu136 and Lys131/Arg132) and two minor
Instituto de Parasitologia y Biomedicina "Lopez-Neyra", (Lys109/Ala110 and Arg90/Asp91) cleavage sites in this
CSIC, Ventanilla, 11, 18001-Granada, Spain; 3Department region. Tryptic peptide mapping indicated that the covalent
of Biosciences at Novum, Karolinska Institutet, Huddinge, dimers of rhIFN-
S-14157 Huddinge, Sweden mainly on account of lateral cross-linking of the monomer
genoveva@obzor.bio21.bas.bg subunits. Antiviral assay showed that the proteolysis
The significance of the C-terminal part of human interferon lowered, while the covalent dimerization completely
gamma (hIFN) for its biological activity was studied by abolished the antiviral activity of rhIFN-.
3‘-end gene mutagenesis. A series of 9 derivative genes
obtained by systemic deletion of 3 codons was constructed ORAL PRESENTATION 3
and expressed in E. coli LE392. It was shown that the yield NONINVASIVE OPTICAL ASSAY OF
of recombinant protein gradually decreased and the SYNTHETIC HEMOGLOBIN FLUIDS
solubility gradually increased with truncation of the C-
terminus. To avoid artifacts related with the imperfect Mehmet D BĠLGĠN
folding of the proteins during purification, the biological Adnan Menderes University, Medical Faculty, Biophysics
activity of the hIFN proteins was measured in clear cell Dept., 09100 Aydın / TURKEY
lysates containing the soluble fractions only. The deletion
mdbilgin@adu.edu.tr
of the C-terminus had a two step effect on both hIFN
antiviral and antiproliferative activities. Whereas the The aim of this study is to measure oxygen saturation and
removal of the last 3, 6 and 9 C-terminal amino acids led to methemoglobin (MetHb) content in synthetic hemoglobin
a gradual increase (up to 10 times) in biological activity of fluids, which contain significant amounts of non-oxygen
hIFN, the deletion of more than 9 amino acids had an carrying MetHb formed by the preparation and during
opposite effect. The truncation of the whole unstructured storage. Conventional oximeters for whole blood do not
C-terminal domain resulted in a 10-fold decrease (but not assay for MetHb.
in a complete loss) in biological activity of hIFN. The Liposome encapsulated hemoglobin (LEH) was used as a
latter was sequestered upon deletion of 24 amino acids, synthetic hemoglobin fluids. Total transmittance and
three of which belonged to the -helical domain F. reflectance of the LEH suspensions {oxyhemoglobin
(OxyHb), deoxyhemoglobin (deoxyHb), MetHb} were
ORAL PRESENTATION 2 measured by spectrophotometer. OxyHb, deoxyHb and
MetHb concentrations were calculated singular value
NON-ENZYMATIC GLYCOSYLATION OF decomposition (SVD) and were compared with known
RECOMBINANT HUMAN INTERFERON- mixtures of [OxyHb]: [MetHb]. Also, diffuse reflection
GAMMA measurements were done in OxyHb, deoxyHb, MetHb, and
1 mixtures of OxyHb-MetHb and OxyHb-deoxyHb.
Roumyana MIRONOVA, 2Toshimitsu NIWA, 1Rositsa
DIMITROVA, 1Maya BOYANOVA and 1Ivan IVANOV The constituent of hemoglobin derivatives analyzed by
1 SVD. The mean deviation of the calculated concentrations
Department of Gene Regulations, Institute of Molecular
Biology, Bulgarian Academy of Sciences, 1113 Sofia, from the ―as mixed‖ values is 2 % for OxyHb and 16 %
BULGARIA for MetHb. In addition, the effect of thermal incubation at
2 40C on LEH was determined. Our experiments stated that
Department of Clinical Preventive Medicine, Nagoya significant loss of OxyHb occurred after 4 hrs and all
University School of Medicine, 466-8560 Nagoya, JAPAN OxyHB was lost after 24 hrs. Also, singlet oxygen
rummy@obzor.bio21.bas.bg quenchers such as imidazole, sodium azide, and ascorbic
Until recently the non-enzymatic glycosylation (glycation) acid were not protective against thermal incubation. But
was thought to affect the proteins of long living eukaryotes radical scavenger (sodium formate) and antioxidant agents
only. However, in a recent study (Mironova, R., Niwa, T., ( -tocopherol) caused protection against thermal effect
Hayashi, H., Dimitrova, R., and Ivanov, I. (2001) Mol. when they were added to the lipid mixture. For example,
Microbiol. 39, 1061-1068) we have shown that glycation for a LEH mixture consisting of 70% OxyHb, 10%
deoxyHb, and 20% MetHb, reflection coefficient was
Turk J Biochem, 2003; 28(3), 62-224 81 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
calculated by SVD and this calculations lead to OCTOBER 14, 2003 – TUESDAY
fractionalOxyHb = 0.7 and fractionalMetHb = 0.20. This
HALL B
approach can employ to determine LEH substitute,
LECTURE 1
containing known amounts of the hemoglobin derivatives.
In conclusion, reflection measurements to analyze for
YEAST CELL WALL PROTEINS:
fractionalOxyHb and fractionalMetHb in hemoglobin fluids LOCALISATION, FUNCTION,
were helped to develop an instrument for performing APPLICATION
noninvasive optical measurements for OxyHb and MetHb. Renata TEPARIģ, Igor STUPAREVIģ, Vladimir MRŠA
Laboratory of Biochemistry, Faculty of Food Technology
ORAL PRESENTATION 4
and Biotechnology, University of Zagreb, Pierottijeva 6,
THE STATUS OF PATIENTS WITH 10000 Zagreb, Croatia
DIABETES MELLITUS MONITORED BY vmrsa@pbf.hr
TURKISH DIABETES SOCIETY Yeast cell wall is composed of structural polysaccharides,
IN DENIZLI / TURKEY glucan, mannan and chitin, and a number of glycoproteins.
Diler ASLAN (1), Nalan AKALIN (1), Gamze CAN S. cerevisiae wall mannoproteins can either be
YILMAZTÜRK (1), noncovalently (Scw proteins - soluble cell wall proteins),
or covalently (Ccw proteins – covalently linked cell wall
Galip YILDIZ (2), Ġbrahim AKKAN (2), Mehmet proteins) bound to glucan. Scws are released from the wall
BOSTANCI (3) by hot SDS, while Ccws are usually extracted by
(1) Pamukkale University School of Medicine Department glucanases although a group of them (so called Pir –
of Biochemistry, 20200, Denizli/TURKEY proteins with internal repeats) can also be released from
glucan by mild alkali treatment.
(2) Turkish Diabetes Society, 20010, Denizli/TURKEY
Different extraction methods reflect differences in
(3) Pamukkale University School of Medicine Dept. of localisation mechanisms of the three groups of proteins.
Population Health Care, 20200, Denizli/TURKEY. For Scws no enzymatic attachment to wall polysaccharides
daslan@pamukkale.edu.tr was postulated and the adsorption may simply be due to
chemical properties of -1,3 glucan which reacts with
Introduction: Optimal control of glycemia in the Diabetes many proteins by hydrogen bonding. Most glucanase-
Control and Complications Trial (DCCT)in USA and the extractable proteins share the localisation mechanism
Prospective Diabetes Study (UKPDS) in U.K. showed the involving the binding of C-terminally attached GPI-anchor
reduction in the incidence and progression of to -1,6-glucan, while the Pir-protein family is anchored
complications of diabetes, and the importance of glycated -1,3-glucan by a so far unexplained reaction
hemoglobin (HbA1c) for guiding therapy for Diabetes involving the characteristic repetitive sequence of these
Mellitus (DM). proteins.
Objective: To determine the status of patients with DM To assess their possible role, cell wall protein mutants like
monitored by the Turkish Diabetes Society (TBS) in SCW4, SCW10, SCW11, and SCW8/BGL2, as well as all
Denizli and the impact of glycaemic status on the four known PIR genes (CCW5, CCW6/PIR1,
complications of DM. CCW7/PIR2/HSP150, and CCW8/PIR3) were constructed.
Methods: The results of the biochemical tests (HbA1c, All mutants were viable, however, some of them like
glucose, cholesterol, HDL-Chol., LDL-Chol., triglyceride, scw4scw10 and scw4scw10bgl2, showed significantly
urea, creatinine, calcium, magnesium, microalbumin) of increased fraction of dead cells in the culture. Additional
848 patients aged 7-85 years admitted between 1999-2003 scw11 mutation suppressed the observed phenotype
to the TDS, and the correlations between HbA1c with the indicating an antagonistic behaviour of Scw11p to Scw4p,
other tests and the complications were evaluated. The Scw10p and Bgl2p.
SPSS program was used in the statistical evaluations. Successive mutations of PIR genes led to changes in cell
Results and Discussion: The HbA1c levels were found as morphology and stability and also to increased mortality.
7.0% (the highest:18.0%) for the 48.3%. There were no investigation showed an increased fraction of cells with
significant differences between parameters measured for more than one bud and in most cases daughter cells still
the two HbA1c groups except HbA1c, glucose (p=0.0001) attached to their mothers stained with methylene blue.
and magnesium (p=0.030) levels. Almost half of the Some potential medical and biotechnological applications
population seems to be monitored properly, but we didn‘t of the obtained results will be discussed.
observe serious examinations for the complications. The
LECTURE 2
results show that more training courses should be arranged
about HbA1c and the complications of DM. The other CATABOLIC AND ANABOLIC AGENTS OF
advantage of this study may be the initiation of the BONE ORGAN-CULTURE
standardization of HbA1c measurements in Turkey.
Sofıc Emın
Keywords: Diabetes Mellitus, glycated hemoglobin,
Diabetes Mellitus and complications, HbA1c
Turk J Biochem, 2003; 28(3), 62-224 82 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
Harvard School of Public Health, Harvard University, inhibition effect. This fenomenon can be observed and
Boston, MA, U.S.A. and Faculty of Science, University of estimated by conducting a calcium analysis.
Sarajevo, Bosnia and Herzegovina
Calvaria of five-day old mice ICR strain, were dissected LECTURE 3
aseptically to encompass part of the frontal bone and most ELECTRO-MANIPULATION OF THE
of both parietal bones. Dulbecco‘s Modified Eagle‘s
Medium containing glucose, glutamine, bovine serum
BIOLOGICAL CELLS BASICS AND
albumin, fraction V, penicillin and streptomycin, were APPLICATIOS
added to each bone culture tube. This medium was serum Mihai RADU
free. Catabolic or anabolic agents were included in the Horia Hulubei National Institute for Physics and Nuclear
medium. The bone culture tubes were incubated in a roller Engineering, Department of Life and Environmental
aparatus for 7 or 14 days at 37 C and oxygenated with 50 Physics, Bucharest – Magurele, PO Box MG-6, R 76900,
% O2, 5 % CO2 and 45 % N2. The media were changed Romania, mradu@ifin.nipne.ro
every 2-3 days and after each change of media, the used
medium from each bone culture tube was analyzed The accumulation of the mobile charges at the interfaces
individually for calcium release from the bone into the between media with different electrical properties
medium. Bones were fixed with formalin and processed for (interfacial polarization) induces the occurrence of an
histological examination. Bones without fixing with electric dipole into a cell exposed to the action of an
formalin were hydrolyzed and analyzed for external electric field. The consequences depend on the
hydroxyproline. Amount of calcium measured from the electric field features (amplitude, frequency, spatial
bone organ culture medium after prostaglandine E2 (PGE2) distribution) and on the cell electrical characteristics
or human parathyroid hormone (h-PTH) fraction 1-34 was (conductivity and permittivity of the media which
included in the medium. Both test substances cause compound the cell). Some of the mechanical effects
calcium release or bones resorption. Reliable and produced by the interaction between the electric field and
consistent calcium resorption from bone using 500 ng/ml the induced dipole led the so called ―electro-manipulation‖
PGE2 or 250 ng/ml h-PTH /1-34) have occurred. of the biological cells. On the other hand, the electrical
potential induced across the cellular membranes is able to
Calcium release from the bone into the medium can be produce local reversible increase of the permeability
observed by using a calcium ion-selective electrode for the (electroporation) allowing to the exogenous chemical
purpose of observation of resorption process. This method species to diffuse into the cell.
is absolutely exact.
The electro-mechanical techniques (dielectrophoresis,
The resorption process in bone organ culture can be electro-rotation and electro-orientation) will be reviewed
measured and evaluated by only measuring the calcium addressing the mechanisms, the theoretical models and the
concentration amount in the medium. Further analysis is applications. The multi-shell model and its use to the
not required. The results, thus, indicate that calcium can be description of the yeast cells mechanical behavior in an
considered as an independent index of bone resorption. external electric field will be discussed. Also other
Reliable and consistent bone formation in bone organ techniques rely on the cell electro-mechanical behavior
culture using ascorbic acid (AA) 150 g/ml or after will be described.
addition 50 ng/ml of bone morphogenetic protein 4 The electroporation of the artificial and natural membrane
(BMP4) have been stimulated. Both substances stimulate will be analyzed in connection with the mechanism of the
osteogenesis. In this case, increased calcium release into phenomenon reversibility. The diffusion of the different
the medium did not occur. kind of molecules will be discussed.
Measuring of calcium release from the bone into the The combination of these effects with other techniques, as
medium using a calcium ion-selective electrode for laser trapping of the cell, produced other complex
observation and estimation of formation process is noninvasive methods for cells investigation and
insufficient. manipulation that will be briefly presented.
A complex method of High Pressure Liquid
Chromatography with Fluorescence detection (HPLC-FD)
of hydroxyproline (biomarker of colagen synthesis in bone OCTOBER 14, 2003 – WEDNESDAY
matrix) or time-consuming and expensive histological HALL B
examination for osteoides observation are necessary.
ORAL PRESENTATION 1
However, both methods can be substituted with simple,
reliable and practical method of biogravimetry, as IMMUNOLIPOSOMES DIRECTED
established by Sofic and Goldhaber in 1998. Significant TOWARD VCAM-1: VEHICLES FOR
increase in calvarial weight at a range of about 50 %, after SPECIFIC DRUG DELIVERY TO
14 days was recorded after adding anabolic agents AA or
BMP4 to the bone organ culture. Significant decrease in ACTIVATED ENDOTHELIAL CELLS
calvarial weight of about 80 %, after 7 days was recorded Manuela VOINEA1, Elena DRAGOMIR1, Ileana
when catabolic agents PGE2 or h-PTH were added to the MANDUTEANU1, Monica CAPRARU2, Dorin
bone organ culture. Neutral red 25-33 g/ml (concentration CAPRARU2, Maya SIMIONESCU1
which is not toxic for bone) added into the medium can
inhibit calcium release in organ culture. Light intensified
Turk J Biochem, 2003; 28(3), 62-224 83 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
1
Institute of Cellular Biology and Pathology “Nicolae Biosensors are defined as an analytical device
Simionescu", Bucharest, ROMANIA, 2 Millitary Hospital incorporating a biological sensing element such as enzyme,
"D.Gerota", Bucharest, ROMANIA tissue, microorganism, cell, and DNA with a suitable
mvoinea@simionescu.instcellbiopath.ro transducer.
Introduction: The use of liposomes as carriers for In this study, two different amperometric biosensors based
selective targeting of drugs and genes to endothelial cells is on catalase immobilized in gelatin-alginate and gelatin--
an attractive strategy in the treatment of cardiovascular carrageenan on a dissolved oxygen (DO) probe covered
diseases. A potential molecular target is vascular cell with a oxygen sensitive teflon membrane, were developed
adhesion molecule-1 (VCAM-1) which is over-expressed for the alcohol determination. Measurements were made by
in vivo by activated endothelial cells (EC) covering the standard curves which were obtained by the determination
developing atheromatous plaque and has an significant role of consumed oxygen level related to alcohol concentration
in leukocyte adhesion to these cells. Taking advantage that according to two reactions catalyzed by catalase given
VCAM-1 can be induced in cultured EC in the presence of below;
inflamatory cytokines and endotoxins, we searched for the Catalase
mechanisms of interaction between activated EC and H2O2 + H2O2 O2 + 2H2O (1)
liposomes targeted to VCAM-1 expressed on the cell
surface. Catalase
CH3CH2OH + H2O2 CH3COH + 2H2O (2)
Materials and methods: Human EC line (EAhy 926)
activated with TNF- were exposed to small unilamellar
liposomes, plain or coupled with anti-VCAM-1 (L-VCAM- The response of the both of two biosensors depended
1) or with irrelevant IgG. For binding studies the cells were linearly on a alcohol concentration range of 0.05 – 0.8 mM
incubated with fluorescently labelled liposomes for 2h at with a response time of two min. For the biosensors
4oC. To follow the fate of liposomes after binding to the developed in order to optimize working conditions some
cell's surface we analyzed the uptake and the optimization studies such as determination of optimum pH,
transmigration of liposomes and the subsequently induced temperature, the most suitable buffer system and
intracellular changes using radioactively labelled concentration were done. In the characterization studies of
liposomes. As methods, flow cytometry, liquid scintilation the biosensors substrate specificity, reproducibility,
counting, fluorescence microscopy and fluorimetry were determination of interference effects of some substances,
employed. operational and storage stability experiments were done.
Results obtained in the optimization and the
Results: The data showed that: (i) liposome coupled to
characterization studies for both two biosensors were also
anti-VCAM-1 bind selectively to activated EC; (ii) the
compared eachother.
immunoliposomes are taken up by specific(e.g. receptor-
mediated endocytosis) and unspecific mechanisms; (iii)
binding of L-VCAM-1 to EC surface induced ORAL PRESENTATION 3
rearrangements of actin filaments and rises in intracellular DETERMINATION OF CHROMIUM(VI) BY A
calcium concentration; (iv) a small percent of liposomes CATALYTIC SPECTROPHOTOMETRIC METHOD
transmigrate into subendothelial space. IN THE PRESENCE OF p-AMINOBENZOIC ACID
Conclusion: The data suggest that VCAM-1 may be an Angelina STOYANOVA
appropriate molecular target for specific delivery of drugs
Higher Medical Institute-Pleven, Department of Chemistry
to activated EC using immunoliposomes.
and Biochemistry & Physics and Biophysics, 5800
This work was supported by the Ministry of Education and Pleven/BULGARIA
Research, National Program VIASAN (Grant nr.
031/2001). astoy@abv.bg
Chromium(VI) is a strong oxidizing agent and possesses
ORAL PRESENTATION 2 high toxicity to humans and animals due to its carcinogenic
DEVELOPMENT OF TWO and mutagenic properties. That is why the determination of
chromium in environmental and biological samples is of
AMPEROMETRIC BIOSENSORS BASED ON great interest.
CATALASE IMMOBILIZED IN GELATIN-
In this work a catalytic spectrophotometric method for the
ALGINATE AND GELATIN-- determination of chromium(VI) is proposed. The method is
CARRAGEENAN FOR ALCOHOL based on the catalytic effect of chromium(VI) on the
DETERMINATION oxidation of sulphanilic acid (SA) by hydrogen peroxide in
Erol AKYILMAZ and Erhan DĠNÇKAYA the presence of p-aminobenzoic acid (PABA) as an
activator.
Ege University, Faculty of Science, Biochemistry
Department, 35100 Bornova-Ġzmir/ TURKEY The reaction was followed spectrophotometrically by
akyilmaz@sci.ege.edu.tr tracing the formation of the reaction product at 360 nm
after 15 minutes of mixing the reagents.
Ethanol is a toxic material which is very often in great need
of being determined in forensic medicine and clinical On the bases of the investigations made, the optimum
toxicology especially as the alcoholic drinks are widely reaction conditions were established:
consumed.
Turk J Biochem, 2003; 28(3), 62-224 84 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
-3 -1
4.0x10 mol l SA, 0.57 mol l-1 H2O2 , 1x10-3 mol l-1 INACTIVATION OF MELANOMA CELLS
PABA and 0.04 mol l-1 acetic acid – boric acid - IRRADIATED WITH GAMMA RAYS AND
orthophosphoric acid buffer solution (pH 6.6), at 50 oC.
LOW ENERGY PROTONS
The linear range of the calibration graph was up to 140 ng
ml-1 and the detection limit was 10 ng ml-1. Interferences Aleksandra RISTIC-FIRA1, Ivan PETROVIC1, Danijela
of Cu(II) and Cr(III) ions were masked. The method was TODOROVOC1, Miroslava VUJCIC1, Lela
applied to the analysis of Cr(VI) in industrial water with KORICANAC1, Sabera RUZDIJIC2, Miroslav DEMAJO1,
recoveries of 95.2 - 104.3 % and a mean RSD (n=6) of Giacomo CUTTONE3
5.6%. 1
Vinca Institute of Nuclear Sciences, Belgrade, Serbia and
Keywords: chromium(VI), catalytic method, sulphanilic Montenegro;
acid, p-aminobenzoic acid, industrial water 2
The Institute for Biological Research, Department of
ORAL PRESENTATION 4 Neurobiology and Immunology, Belgrade, Serbia and
Montenegro;
ALTERED DRUG RESISTANCE AND 3
Istituto Nazionale di Fisica Nucleare, LNS, Catania, Italy.
NEUROLOGIC DISORDERS IN
DROSOPHILA MELANOGASTER WITH A aristic@rt270.vin.bg.ac.yu
DEFICIENT HISTAMINE-GATED Radiotherapy and particularly proton therapy is very
CHLORIDE CHANNEL efficient in eliminating malignant growths, but it is also
very delicate, since healthy tissue surrounding ill tissue
Mladen IOVCHEV1, Plamen KODROV1, Adrian should not be affected at all or very little by the irradiation.
WOLSTENHOLME2, William L. PAK3 and Eugene
The main characteristics of protons, such as their well
SEMENOV1
1
defined range, relatively small lateral scattering, and high
Institute of Molecular Biology, Bulgarian Academy of energy deposition density, just before the end of the range,
Sciences, Sofia 1113, BULGARIA; 2Department of Biology make them particularly suitable when malignant growths
and Biochemistry, University of Bath, Bath BA2 7AY, UK; are deeply embedded or are close to critical organs, where
3
Department of Biological Sciences, Purdue University, there is a high demand to minimize the destruction of the
West Lafayette, IN 47907, USA; esen@obzor.bio21.bas.bg neighbouring and overlaying tissue.
The recent identification and characterization of two genes, In order to obtain better results in eliminating malignant
encoding histamine-gated chloride channel subunits from cells, the aim of this in vitro study was to investigate the
Drosophila melanogaster, has confirmed that histamine is a difference in response of HTB63 human melanoma cells to
major neurotransmitter in the visual system of the fruitfly.
irradiation with either gamma rays or protons considering
One of the cloned genes, hclA, corresponds to ort (ora
dynamics of cell growth. Single irradiation with gamma
transientless), mutations in which affect histaminergic
rays using doses from 2 to 20 Gy exhibited weak
synaptic transmission in the Drosophila visual system. We
identified a mutational change (a null mutation) in the inactivation of human melanoma cells in vitro. The best
genomic and RNA copies of hclA derived from flies with effect, 26% of growth inhibition was obtained after single
the ort1 allele. This correlates with new phenotypes irradiation with gamma ray using dose of 16 Gy. Using the
observed in the mutant strain. We found hypersensitivity same doses of proton irradiation, with energy at the target
to neurotoxins of the avermectin group in both the ort 1 of 22.6 MeV, significant melanoma cell growth inhibition
adult flies and third instar larvae compared to Oregon R was induced. Doses of 12 and 16 Gy provoked growth
wild-type animals. In contrast, the mutation makes male inhibition of 48.9 and 51.2% respectively. Estimated RBEs
and female adults more resistant to treatment with diethyl for inactivation of HTB63 cells ranged from 1.02 to 2.22.
ether, and the animals show substantially prolonged The electrophoretical analyses of DNA samples and flow
recovery from paralysis after diethylether anaesthesia, as cytometric evaluation have shown a small percentage of
well as an impaired recovery from paralysis after apoptotic cells after both types of irradiation.
mechanical shock, as revealed by the bang sensitivity test. The inhibitory effect of protons on melanoma growth, in
The examination of several other alleles ort (with identified contrast to gamma rays, can be explained considering
mutations in hclAs) in the same tests revealed the allele- specific physical properties of protons, especially taking in
specific responses. Altogether, our data give direct account good dose distribution.
evidence that in vivo a HCLA subunit-containing receptor
has a distinct role in the response to general anaesthesia LECTURE 2
and the neurotoxins, as well as indicate that its function is
not limited by the frames of the visual system. LARGE SCALE MACROMOLECULAR
TOPICS: 1) Ion channels and membrane trafficing;
SIMULATIONS BY SYMPLECTIC
2) Metabolic disorders; 3) Molecular structure and function INTEGRATION METHODS
Dušanka JANEŢIĥ
National Institute of Chemistry, Hajdrihova 19, Ljubljana/
OCTOBER 15, 2003 – WEDNESDAY
SLOVENIA
HALL B dusa@cmm.ki.si
LECTURE 1 Among the main theoretical methods of investigation of
the dynamic properties of biological macromolecules, such
as proteins, are molecular dynamics (MD) simulation and
harmonic analysis. MD simulation is a technique in which
Turk J Biochem, 2003; 28(3), 62-224 85 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
the classical equation of motion for all atoms of a molecule Bogazici University, Biomedical Engineering Institute,
is integrated over a finite period of time. The resulting 34342, Bebek - Istanbul, Turkey
trajectory is used to compute time-dependent properties of e-mail: ataakin@boun.edu.tr
the system. Harmonic analysis is a direct way of analyzing
vibrational motions. Harmonicity of the potential function There has been tremendous achivements in brain functional
is a basic assumption in the normal mode approximation imaging techniques, namely in fMRI, MRS and more
used in harmonic analysis. This is known to be inadequate recently in functional optical imaging. However
in the case of proteins because anharmonic effects, which quantification and determination of the physiological and
MD has shown to be important in protein motion, are biochemical mechanisms in the neurovascular system and
neglected. When anharmonic effects are incorporated in neural activation remain to be challenging. For our
quasiharmonic analysis may be applied. In this method, the studies in metabolic disorders and functional optical
MD simulation is utilized to obtain effective modes of imaging, we have implemented a software package which
vibration from the atomic fluctuations about an average uses Aubert and Costalat‘s [1] model of neurovascular
structure. These modes include the anharmonic effects coupling in the brain. The reasons of selecting this model
neglected in a normal mode calculation [1]. are that it is a compact form of the neurovascular coupling,
it is recent, and it takes into account the previous models.
The role of low frequency normal modes involving global
conformation changes and which have been theoretically The model is essentially a coupling model between brain
determined for several proteins is emphasized. Low electrical activity, metabolism, and hemodynamics. This
frequency modes of proteins are particularly interesting model combines the interactions of the following
because theyare related to functional properties. The parameters and mechanisms: (i) cerebral blood flow, (ii)
analysis of these motions in the limit of harmonic intracellular sodium (iii) glycolysis (iv) ATP, PCr, and
dynamics lends insight into the behavior and flexibility of mitochondrial respiration (v) blood–brain barrier
these molecules. The modes presented here include the exchanges and (vi) the Balloon model. The model attempts
lowest modes of Bovine Pancreatic Trypsin Inhibitor to model the relationships between the above-mentioned
(BPTI) [2, 3]. parameters by means of 15 differential equations.
Differential equations‘ parameters were obtained basically
Harmonic analysis also proved useful in developing with the use of fMRI (functional magnetic resonance
efficient symplectic MD integration methods. Symplectic imaging) and MRS (magnetic resonance spectroscopy)
integration methods are often the right way of integrating measurements.
the Hamilton equations of motion. Recent advances in
development of SISM Our software package is developed in Matlab 6.0
environment (in MS Windows XP) and it has a user-
(Split Integration Symplectic Method) and HANA friendly graphical user interface. Basically, it solves 15
(Hydrogens ANAlyticaly) for combined analytical and differential equations and enables us to get the biochemical
numerical solution of the Hamiltonian system based on a responses of the brain under different metabolic conditions.
factorization of the Liouville propagator are presented [4,
5]. We have essentially generated a simulation environment
for neurovascular coupling model of Aubert and Costalat.
SISM and HANA use an analytical treatment of high We plan to use it for the study of metabolic disorders as
frequency motions within a second order generalized leap- well as for the comparison of the responses measured by
frog scheme. The computation cost per integration step for functional optical imaging technique to the simulation
both methods is approximately the same as that of results generated by the implemented model.
commonly used algorithms, and they allow an integration
time step up to an order of magnitude larger than can be ORAL PRESENTATION 2
used by other methods of the same order and complexity.
SISM and HANA have been tested on a variety of ANALYSIS OF MILLISECOND DARK
examples. In all cases they posses long term stability and RELAXATION KINETICS
the ability to take larger time steps while being economical OF CHLOROPHYLL A DELAYED
in computer time.
FLUORESCENCE IN LEAVES DURING THE
The approach developed here is general, but illustrated at INDUCTION PERIOD OF DARK TO LIGHT
present by application to the MD integration of the model
ADAPTATION
system of linear chain molecules and a box of water
molecules. Vassilij GOLTSEV, Ivelina ZAHARĠEVA1, Petar
LAMBREV1, Petko CHERNEV, Chavdar SLAVOV, Ivan
YORDANOV2, Reto J. STRASSER3
OCTOBER 15, 2003 – WEDNESDAY
Department of Biophysics and Radiobiology, Faculty of
HALL B Biology, St. Kliment Ohridski University of Sofia, 8,
ORAL PRESENTATION 1 Dragan Tzankov Blvd., 1164, Sofia, Bulgaria, E-mail:
goltsev@biofac.uni-sofia.bg;
IMPLEMENTATION OF A 1
Institute of Biophysics and 2 Institute of Plant Physiology,
NEUROVASCULAR COUPLING MODEL
Bulgarian Academy of Sciences, “Acad. G. Bonchev” Str.,
Sefer Burak KACAR, Ömer ġAYLĠ and Ata AKIN Bl.21, 1113 Sofia, Bulgaria;
Turk J Biochem, 2003; 28(3), 62-224 86 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
3
Bioenergetics Laboratory, University of Geneva, CH- g/mm (n=10). In addition, it was observed that the slope of
1254 JUSSY – GENÈVE Switzerland the stiffness-force relationship depends on the preload
The contribution of different components of delayed applied to the muscle. When the experimental results
fluorescence (DF) dark decay during the induction period obtained from the portal vein were compared with the
of dark to light adaptation was analyzed. Using theoretical results calculated from the models, it was seen
phosphoroscope fluorometer with high speed digitalization that none of the models could fully represent the portal
a prompt chlorophyll fluorescence signal was registered vein. However Maxwell model can be used if it is assumed
simultaneously with a series of dark relaxation kinetics of that series elastic element of the model has a preload
DF, recorded at different moments during the induction depended stiffness properties.
period. The dark relaxation of DF between 0.35 and 5 ms is This study was supported by the Scientific and Technical
poly-exponential and can be approximated by 3 Research Council of Turkey (SBAG-1206).
components with life-times of about τ1 ~ 0.6, τ2 ~ 3.5 ms
and τ3 ≥ 20 ms. Both the amplitudes and the life-times of ORAL PRESENTATION 4
the DF components drastically changed during the
induction. Тhe contribution of the millisecond components
PURIFICATION AND
with lifetimes 0.6 and 2 – 4 ms predominated during the CHARACTERIZATION OF A PHOSPHATE
first second of the induction period, and later the SPECIFIC TRANSPORTER
amplitudes of the tree components became approximately HYPERALKALINE PHOSPHATASE FROM
equal. The contribution of DF components was highly THERMUS THERMOPHILUS
dependent on registration temperature. At low temperature Anastasia A. PANTAZAKI, Gregoris P. TSOLKAS and
(5 ºC) the main contribution in the fast phase of DF Dimitrios A. KYRIAKIDIS
induction curve had the millisecond component, at high
(38 ºС) – the sub-millisecond and at room temperature (22 Laboratory of Biochemistry, Dept. of Chemistry, Aristotle
ºС) the amplitudes of the both components were University of Thessaloniki, 54124 GREECE
approximately equal. On the basis of kinetic models natasa@chem.auth.gr
describing the redox reactions in the donor and acceptor A soluble alkaline phosphatase from the thermophilic
side of Photosystem II, the participation of different redox bacterium T. thermophilus was purified to homogeneity as
states of the reaction center in the formation of separate a single band on SDS/PAGE with a molecular mass of 40
components of DF dark decay is discussed. kDa. The enzyme exhibited an optimal pH of
Acknowledgments. This work was financially supported by approximately 12.3 and highest activity at 70oC. Among
the Swiss National Science Foundation (SCOPES 2000– the tested divalent cations Ca2+ or Mg2+ cause a small
2003 grant № 7BUPJ062408.00/1). increase in enzyme activity and thermostability as well.
Other cations, EDTA, pyrophosphate, vanadate and
ORAL PRESENTATION 3 molybdate markedly inhibited the enzyme activity. NaF,
tartrate and okadaic acid had a less potent or not at all
DEVELOPMENT OF AN ANALOGUE inhibitory effect. The enzyme hydrolyzed ATP,
MODEL SIMULATING THE PORTAL VEIN phosphoenol-pyruvate and thymidine 3´-monophosphate-p-
nitrophenyl ester (ammonium salt). Its apparent Km for p-
Necla Öztürk nitrophenyl phosphate was 0.1 mM, while for ATP,
Department of Biophysics, faculty of Medicine, Hacettepe phosphoenol-pyruvate, and p-nitrophenyl-3΄-thymidylic
University, Ankara, Turkey acid were 0.006, 0.005, 0.080 mM, respectively. The
It is known that the force generated by muscle cells enzyme was activated approximately 35% and 30% when
p-nitrophenyl phosphate is used as substrate by the
depends on the stiffness of the cross -bridges. In addition,
presence of 40 μM ATP and 100 μΜ of oleic acid,
experiments have shown that the passive tissue mechanics
respectively. N-terminally characterization of protein
modulate the force. Therefore, to understand the interaction
exhibited high degree of similarity with the mature chain of
between the force on the cross bridges and the passive
alkaline phosphatases of PSTS family while partially
tissue mechanics, mechanical models (Maxwell and Voigt)
internal sequence analysis showed that the protein showed
have been used. The present experiment was performed in similarities with ATPases involved. Based on the above
order to find an analog model to simulate smooth muscles data and the enzyme substrates specificities that work as
of the portal vein. For this purpose five different ATPase and phosphodiesterase activity we can conclude
combination of Maxwell and Voigt models were designed that this enzyme belongs to the phosphate specific
and the stiffness-force relations of these models were transporter system (PSTS) which probably participate in
obtained theoretically. Also, the stiffness-force relations of the DNA repair.
the portal vein were obtained experimentally for 7 preload
levels. Stiffness was measured by applying constant
amplitude 5 Hz sinusoidal length perturbations OCTOBER 13, 2003 - TUESDAY
continuously to the contracting muscle preparation. It was HALL C
found that during isometric contractions of the muscle, the
stiffness increased linearly with the isometric force; and LECTURE 1
the slope of the stiffness-force relation is 1.26 0.08
(1/mm) and the line intersects the ordinate at 0.250.13
Turk J Biochem, 2003; 28(3), 62-224 87 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
THREE TYPES OF HOMOCYSTINURIA, A The method of screening for CH involves the
COMMON PROBLEM IN MIDDLE determination of T4 and/or TSH on dried blood spots
collected on the second through fifth days of life. Ten-year
EAST. Pinar T. Ozand MD, Ph. D., Chairman, analysis of our regional screening program for CH (since
Department of Genetics 1991) using primary TSH determination revealed a
King Faisal Specialist Hospital & Research Centre. prevalence of 1/2512 for CH (1/3516 for dysgenesis, and
PO Box 3354, Riyadh 11211, Saudi Arabia. 1/8791 for dyshormonogenesis). Prevalence of transient
Saudi Arabia has an inordinately large number of hypothyroidism was 1/1208. The recall rate was quite high
autosomal recessive diseases. A retrospective study of at 2.8 % reflecting the iodine status of Turkey. Forty
over 1,000 patients indicated that 5 % of them have percent of infants with ectopy, and 20 % of those with
various types of homocystinuria (HCU). In fact this is a dyshormonogenesis had initial serum T4 levels within
common disease in the Middle East area. An average normal limits, in addition to 27.5 % of cases with transient
physician in this part of the world is not well aware of hypothyroidism that needed hormone replacement in early
this disease and missed diagnoses lead to death and life. Hence primary TSH screening is the preferred method
crippling. While successful treatments are available. for screening in Turkey.
Homocystine is an end product of methionine Early detection of CH through newborn screening proved
metabolism. It is toxic and will destroy Fibrillin. This to be one of the great successes of preventive medicine.
leads to the dolicostenomelic features and cataracts of The screening should be oriented to detection of primary
lens. It also damages vascular endothelium causing hypothyroidism. Expected standard for a screening test is a
thrombosis. There are two systems that get rid of it: 1) sensitivity approaching 100 %. However, owing to the
conversion to cystathionine, 2) conversion into element of human error and the potential for biological
methionine by methionine synthetase. The deficiency of variations, no screening test could truly achieve 100 %
cystathionine synthetase, the first pathway, leads to the over long time periods. Simultaneous measurement of both
accumulation of very high levels of homocystine. The toxic T4 and TSH in dried blood spots is the most sensitive
effects start appearing at 4-5 years of age with classic method to that effect, however it is not cost-effective.
clinical picture of HCU. The deficiency of methionine Alternatives are primary TSH, or primary T4 supplemented
synthetase system leads usually to very early by TSH.
symptomatology mainly characterized by failure of the
development of CNS. This usually is caused by the LECTURE 3
deficiency of the cofactors of methionine synthetase,
methylene-tetrahydrofolic (MTHF) acid and EXPANDED NEWBORN SCREENING FOR
methylcobalamine. (Cbl). These latter forms have milder INBORN ERRORS OF METABOLISM BY
elevations of homocystine and very low levels of TANDEM MASS SPECTROMETRY
methionine and are almost always missed. The existence Ġnci KARAARSLAN
of a tandem MS has changed the prognosis of both forms
of this disease. In the last ten years our department didn‘t Düzen Laboratuvarlar Grubu, Ġstanbul/TURKEY
encounter a single thromboembolic phenomenon in incik@duzen.com.tr
classic HCU. We have been able to prevent the delay Screening tests relied on the ―one test-one disorder‖
in CNS development now in five patients with MTHF
concept until the introduction of tandem mass spectrometry
deficiencies.
into newborn screening in the 1990's. Profiling of amino
The metabolic pathways and rationale for treatment of acids and acylcarnitines in a single analysis has enabled
HCU will be discussed. newborn screening programs to expand testing to include
up to 30 treatable inborn error of metabolism(IEM).
LECTURE 2 Besides the increase in the number of diseases covered,
tandem MS has also improved testing from an analytical
SCREENING FOR CONGENITAL point of view. Ġt is very specific and sensitive in its
HYPOTHYROIDISM IN THE NEONATE identification of the compounds. The false positive rates
NurĢen Yordam, MD*, Alev Ozon, MD** are lowered because disorders are identified not only on
* the basis of quantification of metabolites but also by the
Professor of Pediatrics, Hacettepe University Faculty of
screening for a pattern of metabolite abnormalities as
Medicine, Department of Pediatrics, Division of
opposed to screening for a single metabolite and also by
Endocrinology, Ankara, Turkey
measuring metabolite ratios.
**
AssistantProfessor of Pediatrics, Hacettepe University
Between October 2001-August 2003, 12188 newborn (1-10
Faculty of Medicine, Department of Pediatrics, Division of
day old) were screened by tandem MS in our laboratory .
Endocrinology, Ankara, Turkey
%95.5 of the babies were healthy and had normal birth
Congenital hypothyroidism (CH) is a disease long been weight. % 4.5 of the babies either had birth weights less
known to result in mental retardation. It occurs in 1/3000 to than 1500 gr , required neonatal intensive care, or had
1/4000 newborn infants. Early diagnosis and treatment symptoms or familiy history of an IEM. Within the first
prevents brain damage and the ensuing mental retardation. group, three babies with PKU and one with Citrullinemia
Before the era of screening, only 10 % of the affected were identified. In the latter group, we identified 8 amino
infants were diagnosed within the first month of life. acid disorders, 4 urea cycle defects, 7 organic acidemias
and 1 fatty acid oxidation defect.
Turk J Biochem, 2003; 28(3), 62-224 88 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
Within the same period we also screened 1853 patients Health-associated reference values are universally needed
(age 11 days – 14 years old) who had clinical symptoms in clinical chemistry.
associated with IEM. We identified 15 amino acid The aim of this study was to establish the reference
disorders and 13 organic acidemias
intervals of two populations from data obtained by the
The conclusions we can deduct from our experience with mass screening of newborn babies and to demonstrate how
screening for IEM by tandem mass spectrometry are to determine 95 % confidence intervals around the lower
1. The overall frequency of IEM is high in our country and and upper limits of reference values from values that are
newborn screening for these disorders at least in a selected not normally distributed. This experiment shows a way to
high risk group will be cost effective both for the family define the rank numbers for n>1000 and to obtain reference
and for the society in the long run. values with 95% confidence intervals for lower and upper
reference limits.
2. Quite a number of treatable IEM can be rapidly
diagnosed from a very simple sample , namely a dried
blood spot which is both easy to obtain, to tranport and to LECTURE 5
store. This advantage should be made use of for FOETAL BIOCHEMISTRY: BIOCHEMICAL
screening IEM especially in states of emergency and in DIAGNOSIS for FOETUS
cases where laboratories capable of performing advanced
metabolic tests are not readily available. Nezih Hekim
Dr.Pakize Ġ.Tarzi Laboratuarları NiĢantaĢı Ġstanbul
LECTURE 4 Clinical chemistry laboratories are going through a
THE USE OF SPLIT-SAMPLE DESIGN FOR challenge. With the improvement of ―Point of Care‖
system, cinical chemistry laboratories are moving to
PERFORMANCE EVALUATION OF decentralization from centralization, which means that
SCREENING KITS: A REAL LIFE STUDY availability of bed side testing, with the help of
AND EXCEL PROGRAMME FOR automatization and easiness of testing at different
REFERENCE VALUE DETERMINATION OF departments at health centers with decreasing costs clinical
nTSH MEASUREMENT laboratories are moving to new areas of analysis for less
requested but increasing demand tests. Some of these
Prof. Dr. Tijen Tanyalçın (MD, PhD) areas are: Pre implantation genetics, detection of metabolic
Ege University Medical School and Hospital Department diseases of foetus and screening for treatable metabolic
of Biochemistry Bornova 35100 Izmir/Turkey diseases of new born.
Laboratory medicine is an important discipline in health Preimplantation genetics provides the opportunity to detect
care with its remarkable effect on risk assessment, 105.000 single nucleotide polymorphism for many single
diagnosis of health and disease state and especially from gene disease even before fertilization. There are two main
newborn screening approach with its, retest, recall and problems: For applying this method, there is need for
follow-up procedures. This real life trial, emphasizes the conception with assisted reproduction techniques like IVF
and ICSI. Also, at the time, approach to disease detection
need of split sample design evaluation of newly opened test
is to look for mutations on the gene, but there are
kits. Quantitative measurement of phenylalanine and nTSH
variations of mutations in each and every country even in
(neonatal thyroid stimulating hormone) were performed in the same country at different locations and sequence
both of the laboratories. After validation of calibrations analysis is not a screening test. The best approach for the
were performed in the laboratory that used these time seems to analyse amniotic fluid or amnion cells and/or
industrially prepared screening kits for the first time, the their cultured cells during early pregnancy (CVS 15-16
same real newborn blood spot samples were analysed for week) for enzyme or metabolite with classical biochemical
phenylalanine and nTSH measurements in both of the methods. This area is named foetal biochemistry and many
laboratories and the obtained results were compared non biochemistry laboratories are shifting interest to this area.
parametrically and examined by the Deming regression Without any doubt, the diagnosis of the diseases and
graph and by the difference plot. There was no problem abortion after the diagnosis will be within the frame of
with the phenylalanine results, similar results were prenatal rules. Foetal biochemistry will be the starting
obtained for the same blood spot cards from both of the point for in utero genetic treatments in near future. We
laboratories (P=0,496; bias estimation was 0,13). However, have aimed to share our experiment and knowledge on
nTSH values were found to be significantly higher in the foetal biochemistry with our colleagues with this
laboratory that used the nTSH kit for the first time. presentation.
Although the validation of calibration of the nTSH kit was
valid with its own control materials, split sample results OCTOBER 14, 2003 – TUESDAY
showed that there was a significant difference between two
laboratories (P=0,005; bias estimation was 28,6). This HALL C
work implies that acceptable comparability of split sample LECTURE 1
design analysis is strictly needed for testing the analytical
performance of the industrially prepared tests kits and this
AMINO ACID ANALYSIS METHODS –
can be achieved only by certified reference materials. DETAILS AND DIFFICULTIES
Gürsel BĠBEROĞLU
Turk J Biochem, 2003; 28(3), 62-224 89 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
Gazi University, Faculty of Medicine, Department of exchange column, followed by gradient elution. Long
Pediatric Metabolism and Nutrition, 06500, period of investigation is a disadvantage for these methods.
Ankara/TURKEY
Amino acid levels in body fluids are influenced by a
Gurselb@gazi.edu.tr number of factors, such as age, physiological changes,
Disorders of amino acid metabolism constitute an nutritional status, diseases, medications and toxins. Also
important part of inborn errors of metabolism. These the factors such as collection time, transportation and
disorders which are mainly seen in the newborn period and keeping of the samples are very important.
early childhood are characterized by the high levels of one As a conclusion to gain success in the treatment and to
or more than one amino acids in the plasma or urine due to prevent permanent sequels due to disorders of amino acid
the enzyme deficiency. In amino acid metabolism metabolism which makes up a great part of metabolic
disorders, like the other inborn errors of metabolism; early diseases, early diagnosis and treatment is highly important.
diagnosis is very important to prevent morbidity and Also the importance of the amino acid measurement
permanent sequels and also for the success of the techniques is beyond discussion for newborn screening,
treatment. In the light of these points, accurate, sensitive genetic counselling to the family and for the follow-up of
and prompt amino acid analysis in biological fluids is very the patients.
important. Blood, urine, cerebrospinal fluid, vitreous fluid
and amnion fluid are used for the diagnosis of disorders of LECTURE 2
amino acid metabolism.
The analytical techniques for the measurement of amino THE USE OF TANDEM MASS
acids can be investigated in two parts as screening tests and SPECTROMETRY IN NEWBORN MASS
quantitative methods. Screening tests including Guthrie SCREENING PROGRAMS: THE FACTS
test, thin layer chromatography, paper chromatography, BEYOND THE MYTH.
photometric methods and spot tests in the urine. In the
Eyskens François J.M.
recent years a world wide and important technique, that
could screen many metabolic diseases in a single analytical Provinciaal Centrum voor Opsporing van Metabole
step named tandem mass spectrometry has been used for Aandoeningen, Lab Metabolic Diseases, Antwerp,
this purpose. Among the tests those are used for the Belgium.
quantitative measurement of amino acids are capillary Tandem mass spectrometry (TMS) is an analytical
electrophoresis, gas-liquid chromatography, high pressure technique that can be implemented in the analysis of blood
liquid chromatography, ion-exchange liquid spots taken shortly after birth. Rather than testing the
chromatography (amino acid analayzer) and tandem mass blood for the presence of just one compound (e.g.
spectrometry. High resolution nuclear magnetic resonance phenylalanine in the case of phenylketonuria), this
spectroscopy and molecular analysis are also used in amino technique can simultaneously examine a large number of
acid measurements. metabolites (>30 metabolic disorders!) in a single blood
One of the most recent and effective technic of those is spot by ―electronically‖ weigh these molecules. Using this
tandem mass spectrometry. Tandem mass spectrometry is a new technique the screening can be changed from the ―one
very important analytical technique that could determine test-one disorder‖ towards the ―one test-many disorders‖
many metabolic diseases from one blood sample in a very strategy. This technology thus allows a ―sea change‖ in
short time in a single analytical step. Phenylketonuria, newborn screening but it is important that we shouldn‘t
hyperphenylalaninemia, maple syrup urine disease, drown from it: until this moment the experience is limited
tyrosinemia type I and II, homocystinuria, and, except from some defects in fatty acid oxidation (e.g.
hypermethioninemia are the disorders of amino acid MCAD deficiency) and some organic acidemias (e.g.
metabolism those could be determined by tandem mass glutaric aciduria type 1), the expand of the newborn
spectrometry. In addition to amino acid disorders fatty acid screening covering a mass of metabolic disorders is not
oxidation disorders, organic acidemias and urea cycle been studied thoroughly on its efficacy and utility.
defects could also be determined by tandem mass This technique detects well amino acids and acylcarnitines;
spectrometry. at this moment however it is still impossible to screen for
In the newborn period, screening tests were begun by the congenital hypothyroidism or biotinidase deficiency, and
screening of phenylketonuria which is a kind of bacterial there is only a limited experience in screening for
inhibition test progressed by Robert Guthrie. Thin layer congenital adrenal hyperplasia using this technique.
chromatography and paper chromatography are The sensitivity of the screening by TMS is high but the
chromatographic methods those are used for the separation specificity can be rather low with a high rate of false
and determination of amino acids. positives resulting in a high number of retests and recalls:
In quantitative amino acid analysis with high pressure e.g. Belgium (Antwerp, Brussels): retests between 5 and
liquid chromatography the main steps of the method are the 7% in the first two years of screening with TMS; New
pre-column derivatization of the amino acids with South Wales (Australia), Pittsburgh (USA): false positive
phenylisothiocyanate, o-phthalaldehyde and other similar rate of 0.15-0.26%; (for comparison: false positive rate of
compounds, separation in reversed-phase column and phenylalanine screening by an enzymatic method
detection with either ultraviolet or fluorescence detectors. (Quantase, BioRad) is only 0.015%). A broad screening
In amino acid analayzer the main part of the system is ion- program also involves the detection of non-diseases (e.g. 3-
methylcrotonylcarboxylase def. (Germany) and/or atypical
Turk J Biochem, 2003; 28(3), 62-224 90 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
cases whose risk of developing clinical problems is METHODS AND DIFFICULTIES
unknown (e.g. MCAD def. (Australia)). OBSERVED
H. Levy in his editorial on ―Newborn screening by Tandem
Aslan AKSU
Mass Spectrometry: A new era‖ (Clinical Chemistry
(1998);44,12:2401-2) stated that until nowadays screening Akdeniz University, Faculty of Medicine, Department of
services are almost completely controlled by state or other Biochemistry, 07070, Antalya/TURKEY
governmental health departments and that these agencies taksu@akdeniz.edu.tr
generally are not distinguished by their technological
innovation or by their readiness to incorporate new ideas. In this oral presentation, the metabolic disorders of
The great danger in my opinion of the technique of TMS is carbohydrates metabolism will be discussed on basic
that labs that have no experience whatsoever with theoretical concepts and laboratory evaluation point of
screening, will take over the newborn mass screening view. Defective absorption of carbohydrates, glycogen
solely on the base that they possess such an instrument, storage diseases, metabolic disorders causing
without providing a screening program that serves the hyperglycemia and hypoglycemia, mechanism of insulin
community, completely lacking a high technological action, insulin resistance, disorders of fructose and
performance (absence of quality assurance testing) and a galactose metabolism, pentosuria, glucose-6-phosphate
well established structure that allows recovery of all blood dehydrogenase deficiency will be reviewed on this context.
cards and having good contacts with treating physicians.
Last but not least one should not ignore the higher costs of
TMS screening in comparison with the current used OCTOBER 15, 2003 – TUESDAY
techniques (literature: $ 0.7-20; own experience: $ 6 HALL C
supplementary costs/screened newborn).
LECTURE 1
Conclusion: Mass screening of newborns should stay
centralised in the screening labs that have the most DYSREGULATION OF P450c17 ENZYME IN
experience and the best performance; only these labs can POLYCYSTIC OVARY SYNDROME
implement new techniques in a way that newborn
Fahrettin KELEġTĠMUR
screening remains on a high level.
Erciyes University Medical School, Kayseri, Turkey.
Screening is not just a job, it is a dedication.
fktimur@erciyes.edu.tr
LECTURE 3
Polycystic ovary syndrome (PCOS) is the most common
DISORDERS OF CARBOHYDRATE reproductive endocrinopathy of women in their
METABOLISM: CLINICAL APPROACH, childbearing years. PCOS is estimated to affect 5 % of
women of reproductive age and it is associated with higher
FUTURE PROSPECTS OF DIAGNOSIS AND
rates of cardiovascular risk factors and cardiovascular
TREATMENT disease. Current data demonstrate that type 2 diabetes,
Prof. Dr. Benal Büyükgebiz hypertension and hyperlipidemia are more frequent among
Pediatri Anabilim Dalı, Dokuz Eylül Üniversitesi Tıp women with PCOS which is a form of functional ovarian
Fakültesi hyperandrogenism. On the other hand, functional adrenal
hyperandrogenism (FAH) which is characterized by
Carbohydrates are the body's sugar source. Sugars used to hyperresponsiveness of adrenal androgen production to
provide energy for the body include glucose, sucrose, ACTH is also seen in PCOS women. The most likely cause
fructose among many others. Some sugars need to be of increased androgen production by both the ovaries and
broken down, usually by enzymes, before they can be used the adrenals is abnormal regulation of 17-hydroxylase and
by the body. If the enzymes needed are not present (usually 17-20 lyase activation of P450c17 enzyme. Insulin
due to an inherited disorder), these sugars can build up and resistance, at least in part, is responsible for the elevated
cause problems. The type of problem depends on the sugar androgen production. Recent data suggest that amelioration
involved and the localization of the enzyme defect. Most of of insulin resistance may lead to improved
the inherited disorders of carbohydrate metabolism fall into hyperandrogenism.
a few broad clinical syndromes. Hepatomegaly,
convulsions, hyperbilirubinemia, cataract, mental LECTURE 2
retardation, diarrhea, episodic lactic acidosis from early
infancy, failure to thrive, and hypotonia are most common NEUROENDOCRINE SYSTEM AND
signs and symptoms. The demonstration of defective OBESITY
enzyme activity must serve as the basis of diagnosis and
Üstün Korugan.M.D.
treatment.
Prof.Med.Univ of Istanbul
LECTURE 4
Energy expendıture and inhibition of appetite are increased
DISORDERS OF CARBOHYDRATE by Hypotalamic stimulii.
METABOLISM: BASIC CONCEPTS, The hormon LEPTIN which is produced in the fat tissue
EVALUATION OF LABORATORY binds to its own spesific receptors in the Hypothalamus and
causes the inhibition of appetite and the energy
Turk J Biochem, 2003; 28(3), 62-224 91 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
loosing.One the other hand a negativ feed back system mutations, and hepatic lipase-lipoprotein lipase, lechitine
operates between Cortisol cholesterol acyl transferase activities are being used to
Leptin and Insulin.Insulin and Cortisol stimulate the determine the etiopathogenesis of lipoprotein metabolism
production and the secretion of Leptin.In turn Leptin disorders.
inhibits the secretion of Insulin. The second category of the lipid metabolism disorders is
NPY is another Hypothalamic neuropeptide which composed of mitochondrial fatty acid oxidation defects.
These disorders are caused by a group of enzyme
stimulates the appetite and promotes the energy
deficiencies and transport defects, and clinically
expenditure in contraste to LEPTIN.NPY stimulates the
characterized by hypoglycemic-hypoketotic coma, induced
Hypothalamico-Hypopyseal axis,causes the increase of
by fasting. In acute phase of these disorders, serum
ACTH and Cortisol.In turn NPY production is inhibited by
electrolytes, glucose, ammonia, transaminase levels are
ACTH and LEPTĠN whose production and secretion is routine screening tests, while carnitine-acylcarnitine levels,
stimulated by Cortisol.On the other hand NPY stimulates and acylcarnitine profiles, urinary organic acid analyses by
the secretion of Insulin, and eventually the production of GC-MS, measurements of enzyme activities, and mutation
LEPTIN.LEPTIN inhibits the secretion of NPY. analyses are required in determining the etiology. With its
Apart from these, two groups of neuropeptideswhich are significant incidence detected in these patients, and similar
called OREXIGENIC and ANOREXIGENIC take place in frequency as phenylketonuria in North Europe, MCAD
the Hypothalamus.LEPTIN inhibits the OREXIGENICS deficiency is thought to be included in newborn screening
and stimulates the others. programs. It is advised to check acylcarnitine levels,
especially octanoylcarnitine by tandom-MS, in blood
Besides the Hypothalamic peptides, intestinal peptides play samples taken for screening of phenylketonuria and
roles in the control of appetite.OREXINES and neonatal hypothyroidism.
COLESYSTOKININ are the intestinal substances which
can be found olso in the brain,enhances and supress feeling LECTURE 5
of hunger respectively.
The transfer of Phenylalanine into the CSF promotes the THE GLYCOSPHINGOLIPIDOSES: FROM
production and secretion of SEROTONIN which inhibits DISEASE TO BASIC PRINCIPLES OF
the appetite and especially CARBOHYDRATE METABOLISM
CRAVING.INSULIN which is secreted as a result of H. Asuman ÖZKARA
carbohydrate consumption promotes the transfer of
Phenylalanin into the brain tissue. Hacettepe University, Faculty of Medicine, Department of
Biochemistry, 06100 Ankara/TURKEY
LECTURE 3 ozkara@hacettepe.edu.tr
The glycosphingolipidoses are a set of diseases that are
IS OBESITY AN INFLAMMATORY caused by defects in the lysosomal degradation of
DISEASE? glycolipids derived from the plasma membrane.
Candeğer Yılmaz Catabolism of these lipids contains enzymes and activator
proteins.
Over the past decade, biochemical and molecular genetic
LECTURE 4
studies of sphingolipidoses have expanded our
LABORATORY DIAGNOSIS IN understanding of underlying metabolic principles of these
LIPOPROTEIN AND OTHER LIPID diseases and their genes. A new lysosomal digestion model
was developed and mechanisms of glycosphingolipids
METABOLISM DISORDERS hydrolysis within the lysosome was understood. The
ESKANDARI H. G. discovery of sphingolipid activator proteins was an
Dept. of Biochemistry, Medical Faculty. Mersin University, important factor in this process. By investigating the
molecular basis of the diseases, basic principles of storage
Mersin, Turkey
disease pathology begin to understood and several
Disorders of lipid metabolism will be discussed in two mechanisms were described in the pathogenesis. However,
major categories. One of them is composed of frequently our understanding of pathogenesis in these diseases is
seen lipoprotein disorders in clinical biochemistry incomplete. The generation of mouse models and
laboratories. Different disorders of lipoprotein metabolism sphingolipid research on cell signaling will help to
are triggered by apolipoproteins, enzymes, and lipoprotein investigate the pathophysiology and have facilitated the
receptors, some clinically characterized by development of new and promising therapeutic strategies
hyperlipoproteinemia. These disorders can be classified for these diseases, most of which are not treatable at
under seven titles according to Frederickson and WHO. present. Currently few options exist for therapy. One of
Hypolipoproteinemias which are not seen as frequent as these is enzyme replacement therapy (ERT) that has been a
hyperlipoproteinemias are also types of lipoprotein highly effective therapy in type 1 Gaucher disease and
metabolism disorders. Routine tests such as total more recently has been undertaken in Fabry disease. ERT
cholesterol, triglyceride, HDL cholesterol, LDL is not beneficial to the neuronopathic form of
cholesterol, lipoprotein (a), apo AI, and apo B, and spesific glycosphingolipidoses. Gene therapy holds considerable
tests such as HDL subfractions, lipoprotein (a) isoforms, promise for this family of diseases and evaluation in mouse
apo E polymorphism, apo B-3500-apo CII-apo CIII models is a major way forward in evaluating different gene
delivery systems and evaluating efficacy. Small-molecule
Turk J Biochem, 2003; 28(3), 62-224 92 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
drugs have recently emerged as candidate therapeutics for HYPERHOMOCYSTEINEMIA AND LVH IN
juvenile and adult forms of glycosphingolipidoses. The NIDDM
approach is to use inhibitors of glycosphingolipid
biosynthesis and thereby reduce the number of Hülya Yılmaza , Bedia Ağaçhana, H. Arzu Ergena, Zeynep
glycosphingolipid molecules the cells degrade. These drugs ErmiĢ Karaalib, Turgay Isbira
a
have been evaluated multiple mouse models of Institute of Experimental Medical Research, Department
glycosphingolipidoses, including those with brain of Molecular Medicine, University of Istanbul, Istanbul,
involvement. In all cases efficacy has been demonstrated. TURKEY; b Taksim State Hospital , Department of
It is interesting to reflect that diseases that glycolipid Internal Medicine, Taksim, Istanbul, TURKEY
catabolism helped unravel the basic biochemistry of Elevated plasma concentrations of homocysteine (Hcy) as
glycolipids and this knowledge, in turn, has led to new a risk factor of coronary artery disease (CAD), essential
therapies for these diseases. The science has therefore gone hypertension and diabetic target organ damage.
full circle from disease to basic biochemistry to disease Methylenetetrahydrofolatereductase (MTHFR) gene
therapy. C677T polymorphism is associated with
hyperhomocysteinemia. This study was designed to
investigate an association of MTHFR C677T
13.10.2003 P1 polymorphism with homocysteine levels and diabetic
complications in the Turkish population.
APOE AND PON 55/192 POLYMORPHISM Our study was carried out in 249 patients with type II
AND EFFECTS ON PON ACTIVITY AND diabetes mellitus (T2DM) (102 men, 147 women) and 214
LIPIDS IN NIDDM healthy volunteers as controls (91 men, 123 women).
Serum Hcy levels were measured in the fasting state by
Bedia AĞACHAN1, Hulya YILMAZ1, Zeynep
KARAALI2, Turgay ĠSBĠR1 immunological assay. MTHFR C677T genotypes were
1
determined by PCR, RFLP techniques.
University of Istanbul, Institute of Experimental Medical
Research, Department of Molecular Medicine, Istanbul, No differences were observed in the distribution of
Turkey; 2Haseki State Hospital, Department of Internal MTHFR genotypes or allele frequencies in cases versus
Medicine, Istanbul, Turkey controls. In the T2DM and control groups, The
homozygous mutant genotype (T/T) had the highest
tisbir®superonline.com homocysteine levels compared to wild (C/C) and
Objective: Paraoxonase (PON1) and Apolipoprotein E heterozygous mutant (C/T) genotypes (p0.05). Plasma Ca t
control. Plasma was obtained in fasting state between 8 to (p0,05),
patients with essential hypertension and twenty-eight NO (p0,01); the Utku, H1, Miller Lisa M2
concentration of P was for 79,85% higher in the pubertal 1. Institute of Nuclear Sciences, Hacettepe University,
persons compared with the persons over 65 years Beytepe, Ankara, Turkey
(p>0,001).
2. National Synchrotron Light Source, Brookhaven
Conclusion: The obtained results suggest relationship National Laboratory, New York, USA
between concentration of Ca and P with the age. The older
persons have significant lower level of Ca and P compared Fluorescence-assisted synchrotron IR microspectroscopy
with the young persons. permits us to identify target proteins with fluorochromes or
fluorescent antibodies and simultaneously determine their
P24 secondary structure with IR micro-spectroscopy.
CONJUGATION OF DIFFERENT Fluorescence microscopy may be used to identify amyloid
SUBSTRATES WITH GLUTATHIONE plaques and tangles in the brain and other tissues of
CATALYSED BY GLUTATHIONE-S- control. We have incorporated protein structure methods
into Matlab to generate routines for image processing.
TRANSFERASES T1-1 Once the desired structural images are obtained, correlation
Yasemin AKSOY1, Anna-Karin LARSSON2, Bengt analysis can be performed with the fluorescence
MANNERVIK2 microscopy images.
1
Hacettepe University, Faculty of Medicine, Department In order to generate IR images, the IR data was reduced to
of Biochemistry, 06100, Ankara/Turkey, a relatively compact description through cluster analysis.
2
Uppsala University, Biomedical Center, Departmentn of The principal function of clustering is to display so that the
Biochemistry Uppsala/Sweden influences or causes in arriving a pathogenic state might be
predicted. Desired function of clustering is to reveal the
yaseminb@hacettepe.edu.tr protein structures in an infected tissue. Result of the cluster
Glutathione S-transferase (GSTs; EC 2.5.1.18) is a analysis can contribute directly to classification schemes. If
detoxifying enzyme catalyzing the conjugation of the grouping suggested by the cluster analysis is to be
glutathione with a variety of electrophilic substrates that adopted for operational use, then it may become the basis
can be either exogenous or endogenous. In mammals 8 for classifying new observations. The developed software
different classes (alpha, kappa, mu, omega, pi, sigma, theta provides Linear Discriminant Analysis (LDA) option to
and zeta) of soluble GSTs have evolved with members that perform classification based on the output of the cluster
promote the detoxification of many structurally different analysis.
electrophiles. The evolution of proteins for novel functions
involves point mutations and recombinations of domains or
structural segments. P26
Alkyl halides, epoxides and benzyl halides are substrates of POSSIBLE ANTIOXIDANT EFFECTS OF
GSTs. Substrates which were worked are industrial CALCIUM CHANNEL BLOCKER IN
intermediates, laboratory reagents. It behaves as alkylating HYPERTENSION PREGNANT WOMEN
agents. Reports have shown them to cause the respiratory
Didona UNGUREANU1, Mihaela MAFTEĠ1, Cristiana
and dermal toxicity in animals and humans. It has also
Filip1, Nastasia GHEORGHĠTA1, Alexandrina CABA2
been reported to be carcinogenic in experimental models.
Thus, the wide-spread use of these aliphatic epoxides, 1.University of Medicine and Pharmacy “Gr.T.Popa”,
halides is of great concern in human health problem. Biochemistry Department, 700115, Iasi, Romania,
didonaungureanu@yahoo.com
In our study, we purified GST T1-1 (h T1-1) from E.coli.
GST activities towards 1,2 Epoxy-3-(4-nitrophenoxy)- 2.”Cuza-Voda”Hospital, First Clinic, Iasi, Romania
propane(EPNP) and 4-nitrophenethyl bromide (NPB), Arterial hypertension in third trimester pregnant women is
styrene 7,8 oxide, acrylonitrile, benzyl bromide benzyl defined as values higher then 140/85 mmHg, one of the
chloride, epichlorohydrin, glycidol were measured. pathogenic mechanisms involved being intracellular
Reactions of substrates with glutathione were measured by calcium accumulation. Nifedipine- as a calcium channel
following the disappearance of glutathione. Glutathione blocker- in pregnant women hypertension will prevent
was measured colorimetrically using Ellman‘s reagent calcium accumulation and thus, will restore blood pressure.
(DTNB method). Results of DTNB method were compared
with EPNP results. Activities were expressed as micromole
Turk J Biochem, 2003; 28(3), 62-224 102 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
The study was carried out on 25 third trimester pregnant P28
women, aged between 25 and 35 years old, diagnosed with
pregnancy induced hypertension (PIH), and treated with
RESPONSE OF BARLEY SEEDLINGS TO
nifedipine. The results were compared with a control UV-B RADIATION AS AFFECTED BY NACL
group- 20 third trimester normal pregnant women. At both Irena GRIGOROVA, Ivanka FEDINA, Katya
groups we dosaged enzymatic and nonenzymatic GEORGIEVA
antioxidants parameters: superoxide dismutase (SOD), Institute of Plant Physiology, Bulgarian Academy of
glutathione peroxidase (GPx), catalase (CAT), glutathione Sciences, Akad. G. Bonchev str., bl. 21, Sofia 1113,
(GSH) and uric acid (UA). As marker of lipid peroxidation Bulgaria; Djelepova@yahoo.com
we used malondialdehyde (MDA). Under Nifedipine
treatment antioxidant enzymes activities and GSH The effect of pre-exposure to NaCl on barley seedlings
increased (p 3). The production, distribution, and use of electricity promote
former group has been shown to be associated with a cancer or initiate other health problems?".This study
higher risk of breast cancer than the latter group. Arginase, evaluated the possible effect of a sinusoidal 50 Hz
a cytoplasmic enzyme, catalyses the hydrolysis of L- magnetic field (1.35 mT) on the sperm count, testes, liver,
arginine to urea and ornithine in the last step of kidney and brain histopathology, malondialdehyde (MDA)
mammalian urea cycle. Arginase also play an important
concentration of the tissue under investigation, p53
role in the synthesis of polyamines through ornithine.
immune reactivity of bone marrow and some trace
Polyamines were shown to be a cell promoter and their
elements in blood of rat. Sixteen Sprague-Dawley male rats
levels have been found to be higher in malignant lesions. In
were separated into two groups of eight, sham exposed
addition, arginase activities were found to be higher in
several carcinomas including breast, colorectal, stomach (control) and experimental. The rats in the experimental
and prostate. Therefore, presence of arginase and ornithine groups were exposed to Extremely Low Frequency
in breast cyst fluid may help to develop breast carcinoma Magnetic Field (ELF MF) 2 hr/day/ for 2 months (7 days a
from the gross cystic disease of breast. In this study, we week). Eight rats of sham group were treated like
have determined the arginase enzyme activities and experimental group except ELF MF exposure. The Mann-
ornithine levels in breast cyst fluid samples. Arginase and Whitney U-test was used for statistical comparisons of
ornithine levels were measured spectrophotometrically groups. No statistically significant alteration in any
using thiosemicarbazide diacetylmonoksime urea and endpoints was noted except Mn+2, concentrations
Chinard methods respectively. There was no istatistically (p0.05) and
serum total sialic acid levels were found to be 62.30±11.80 in control group (r= -0.272, p>0.05). As a result, we may
mg/dl in patients with benignant skin tumors, 68.31±11.27 report that serum TSA and ceruloplasmin levels are
mg/dl in patients with malignant skin tumors and elevated at 24h post-infarction in patients with acute
51.40±4.26 mg/dl in control group. There was a significant myocardial infarction and an increased output of
difference between serum total sialic acid levels of control ceruloplasmin from the liver cannot be only factor
group and patients with benignant or malignant skin responsible for an increased serum TSA concentration
tumors (p0.05;tHCYmol/l (15.83.23 v.s oxidative stress activation and rising of reactive oxygen
14.723.11) p>0.05. There was statistically high species and lipid peroxidation toxic products.
correlation between Lp(a) concentration at baseline and 12
months later r=0.936 p0,05). In spite of the significant decrease
spectrophotometrically. It was shown an increase
in plasma viscosity, there was no significant improve in
concentrations of stable aldehyde lipid peroxidation
fibrinogen levels those which one of the determinants of
products in the medium by 2.88-6.76 - fold. Under these plasma viscosity. Therefore, we considered that
conditions, the iodine uptake by thyrocytes was almost Atorvastatin‘s improving effects on lipid profile could have
completely inhibited within 2 hours. A 31.1% decrease in positive effects on other determinants of hemorheological
TPO activity was also found in 2 hours, at Fe2+/ascorbate parameters.
concentration of 0.1 x10-4M. At higher concentrations,
P89
TPO was inhibited by 30% after 5 hours and by 61.55%
after 8 hours. The TPO inhibition and iodine uptake were ERYTHROCYTE MEMBRANE PROTEINS
reversible since after 24 hours the enzyme activity was AND LIPID COMPOSITION IN
recovered to the control values. The addition of the 1x10-2 ATORVASTATIN ADMINISTERED
- 1x10 -4 M H2O2 concentrations leads to 24-h inhibition PATIENTS WITH
of iodine uptake and a decrease of TPO activity by 17.23 - HYPERCHOLESTEROLEMIA
33.4%. The data obtained suggest pronounced sensitivity BIKMAZ PS1, ALBENIZ I2, BIKMAZ B3, GOKKUSU C4,
of thyroid hormone synthesis in the thyroid and oxidative TAMER S1
stress activation. The disturbed iodine uptake, as well as its 1 Department of Physiology, Istanbul Faculty of Medicine,
oxidation and organification by thyrocytes seem to be the Istanbul University, Findikzade, Istanbul / Turkey
most important mechanism of thyroid function impairment 2 Department of Biophysic, Istanbul Faculty of Medicine,
under the action of an unfavourable ecologic factor. The Istanbul University , Findikzade, Istanbul / Turkey
research was supported by a grant from the Belarusian 3 Department of Neurosurgery, SSK Okmeydanı
Foundation for Fundamental Studies (Grant B01-343). Educational Hospital, Okmeydani, Istanbul / Turkey
4 Department of Biochemistry, Istanbul Faculty of
P88 Medicine, Istanbul University, Findikzade, Istanbul /
THE EFFECT OF HMG-COA REDUCTASE Turkey
sbikmaz@hotmail.com
INHIBITION ON HEMORHEOLOGICAL
PARAMETERS Although HMG-CoA (3-Hydroxy-3 Methyl Glutaryl Co
enzyme A) reductase inhibitors are known as significantly
BIKMAZ PS1, BIKMAZ K2, TAMER S1, YIGIT R1 effective in reducing plasma and cholesterol levels, not
1
Istanbul University, Istanbul Faculty of Medicine, many studies concerning the relationship between
Department of Physiology, Findikzade, Istanbul / Turkey membran lipid and protein levels are available. In our
2
study, we aimed to examine the changes in erythrocyte
SSK Okmeydanı Teaching Hospital, Department of membrane proteins and lipid composition in
Neurosurgery, Okmeydani, Istanbul / Turkey hypercholesterolemic patients administered with
sbikmaz@hotmail.com atorvastatin, a HMG-CoA (hydroxy methyl glutaryl Co
enzyme A) reductase inhibitor. Therefore, 20 patients
3-Hydroxy-3 Methyl Glutaryl Co-enzym A (HMG-CoA) whose were enrolled in Okmeydani Training Hospital
reductase which is participated in cholesterol synthesis General Internal Diseases Clinic, were included the study.
starting with the Acetyl-coenzym A in liver cell catalyse The patients had no clinical symptom except
the conversion of HMG-CoA to Mevolanat, which is a hyperlipidemia. The hyperlipidemic patients were treated
velocity limiting stage in cholesterol biosynthesis in with orally administered atorvastatin (20 mg/day) during
human. HMG-CoA reductase inhibitors inhibit HMG-CoA 12 weeks. At the end of this time period, the lipid
reductase enzym competitively thus reducing the composition in plasma and erythrocyte membranes (RBC)
cholesterol and lipoprotein levels in liver cells. As a result, was determined using enzymatic methods whereas
they decrease LDL cholesterol and total cholesterol level membrane protein levels were determined by SDS-PAGE
by reducing lipoprotein synthesis and also increasing the electrophoresis method. When the findings were compared
entrance and destruction of the lipoprotein containing Apo- to healthy control group (n=15), a significant reduction
B to the liver cells and the other cells. It is reported that, in (p30 mg/dl (
activity of -galactosidase from Aspergillus orysae. RR= 0.18, 95%CI: 0.10-0.32, p = 2 x 10-5): HTA
Enzymatic synthesis of oligosaccharides and (RR=0.30, 95% CI: 0.19-0.48; p = 1 x 10-5): LMW-S1
alkylglycosides via transglycosylation reaction of lactose apo(a) isoform (RR=7.04, 95%CI: 1.40-35.40, pS4:(RR= 2.59; 95%CI:1.28-5.21, p30mg/dl and
tetrasaccharide and new, different from the lactose plasma total/HDL Ch. ratio (4.5-5.8), then in those with
Turk J Biochem, 2003; 28(3), 62-224 131 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
plasma Lp(a) levels5.8 Note. Values are means±SEM, enzymatic activities expressed as U/mg protein,
*
PS4 apo(a) isoforms, HTA and ethanol intake.
combination of increased Lp(a) levels and total/HDL
cholesterol ratio increase the risk for the development of
atherosclerosis in patients with NIDDM. P105
MEAN CORPUSCULAR VOLUME AND
P104 ENZYME ACTIVITIES IN ALCOHOLIC
LIVER DISEASE
EFFECTS OF PROLONGED ETHANOL
CONSUMPTION ON LIPID PEROXIDATION Lejla BEGIģ, Aida ARNAUTOVIģ1
AND ANTIOXIDANT ENZYMES IN RATS Medical Faculty University of Tuzla, Univerzitetska 1,
75000 Tuzla, Bosnia and Herzegovina
Diana DINU, Marina Tamara NECHIFOR and Gheorghe
1
STOIAN Internal Clinic UKC Tuzla
University of Bucharest, Faculty of Biology, Splaiul lejla_begic@hotmail.com
Independentei 91-95, Bucharest/ROMANIA Harmful alcohol use is a huge socio-economic problem.
nemar@bio.bio.unibuc.ro Clinical features of alcoholic liver disease are: fatty liver,
alcoholic hepatitis and cirrhosis. If alcohol problems are
Ethanol consumption may result in an increased oxidative
recognized at an early stage, a physician may prevent
stress with formation of lipid peroxides and free radicals.
further development and progression of disease. The aim of
Antioxidant enzymes are very important scavengers of
our work was the evaluation of MCV (mean corpuscular
oxygen radicals in the cell. Thus, the purpose of this study
volume), ALAT (alanine aminotransferase), ASAT
was to examine the effects of oxidative stress induced by
(aspartate aminotransferase) and GGT (gamma-
long-term ethanol consumption on main antioxidant
glutamyltransferase) assays in alcoholic liver disease, and
enzymes (superoxide dismutase, catalase and glutathione
their application as markers in chronic alcoholism.
peroxidase) in rat liver and kidney. We have also measured
the level of malondialdehyde content as an indicator of The investigation included 54 patients with reliable
lipid peroxidation process. anamnesis data about chronic alcoholism. By needle
biopsy and ultrasound of liver, examinees were classified
Wistar rats were given ethanol (2g/kg of body weight)
into 5 groups. The MCV is determined by automatic
daily by intragastric infusion. Rats were sacrificed in
method based on alteration of impedance. ALAT, ASAT
groups of 16 (8 ethanol treated and 8 controls) after 10 and
and GGT activities in serum are determined by
30 weeks of treatment. Malondialdehyde was determined
spectroscopic methods.
by a colorimetric test with thiobarbituric acid. Superoxide
dismutase activity was determined by using nitroblue In total sample, the biggest group was composed of
tetrazolium as a detector of superoxide anions. Catalase patients suffering cirrhosis (19) followed by groups of
activity was measured by H2O2 disappearance at 240 nm. patients suffering hepatitis (16), steatosis (12),
Glutathione peroxidase activity was performed following hepatocellular carcinoma (6) and fibrosis (1). As many as
NADPH oxidation at 340 nm. Statistical analysis was 68.52% of all patients have had increased values of MCV,
carried out using Student's t test. 75.9% had increased levels of ASAT, 55.6% had increased
The content of lipid peroxidation products estimated as levels of ALAT, and 90.7% had increased GGT activity.
malondialdehyde does not present significant modification The ratio ASAT/ALAT in our patients was 1.88.
in the liver and the kidney of ethanol-treated rats. The
activities of antioxidant enzymes were increased after Based on our investigation we can conclude that the GGT
prolonged ethanol consumption. The results are presented level in serum and the ASAT/ALAT ratio are valuable
in the table to follow. indicators of chronic excessive alcohol intake. The major
shortcomings of the GGT as a marker of excessive alcohol
Liver Kidney consumption are lack of both sensitivity and specificity.
10 weeks 30 weeks 10 weeks 30 weeks
Numerous other disorders and drugs can elevate the GGT
Superoxide
dismutase and produce false positive results. The ASAT/ALAT ratio
Control 113.12±9.4 93.6±8.3 2.76±0.12 2.52±0.11 is better marker of alcoholic liver disease than separate
Ethanol 139.3±12.1* 116.3±9.5* 3.04±0.31 2.82±0.16
Catalase
serum levels of ASAT and ALAT.
Control 890.3±71 812±60 194±11 176±15
Ethanol 1083±102* 1089±95* 201±14 206±9*
Glutathione P106
peroxidase
Control 0.92±0.08 1.17±0.08 0.034±0.0016 0.03±0.002
Ethanol 1.44±0.13* 1.63±0.15* 0.045±0.0021* 0.044±0.0023*
Turk J Biochem, 2003; 28(3), 62-224 132 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
TRANSCRIPTIONAL INACTIVATION OF Biosensors have found promising applications in various
fields such as biotechnology, food and agriculture product
THE DNA-REPAIR GENE MGMT IN
processing, health care, medicine and pollution monitoring.
PATIENTS WITH ORAL CANCER The combination of oxidoreductases and amperometric
Tatjana DRAMIģANIN1, Koviljka KRTOLICA1, Nasta electrodes is by far the most commonly studied biosensor
concept and through various strategies the enzyme reaction
DEDOVIģ1, Nikola TANIģ2, Dragana TRIFUNOVIģ1,
can be easily followed and sensitively measured by
Miodrag GUŢVIģ1 and Bogomir DIMITRIJEVIģ1 electrochemical means. Laccases (benzenediol: oxygen
1
Institute “Vinca,” P.O. BOX 522, 11001 Belgrade, Serbia, oxidoreductase, E.C. 1.10.3.2) are copper containing
1 oxidoreductases produced by higher plants and
Institute for Biological Research “Siniša Stanković,” microorganisms, mainly fungi and have wide substrate
11001 Belgrade, Serbia specificity and a great potential for the determination of
email: tatjana@vin.bg.ac.yu phenolic compounds which are highly toxic, carcinogenic
and allergenic and due to their toxic effects, their
Alterations in DNA methylation has been proposed as a determination and removal in the environment are of great
central phenomenon underlying the neoplastic importance.
transformation. Generally, hypermethylation is one of the In this work, thick film biosensors containing Trametes
most important epigenetic mechanisms responsible for versicolor (TvL) laccases were developed for the
inactivation of gene transcription. Consequently, determination of phenolic compounds and the
methylation of CpG islands in promoter region of DNA measurement was based on oxygen consumption in relation
repair genes, such as MGMT, will result in loss of MGMT to analyte oxidation. The electrodeposited organic
polymer; polyaniline was used as a matrix for the
protein responsible for the correction of G to A point
immobilization of biological compounds. The systems
mutations. These mutational events, the consequence of were calibrated for different phenolic substances. A
MGMT hypermethylation, may generate genomic linearity was obtained in concentration range between 0.4
instability associated with promotion and / or progression and 6.0 M phenol, 0.2 and 1.0 M catechol, 2.0 and 20.0
of neoplastic phenotype. M L-DOPA, respectively in the response time of 300 sec.
Our aim was to determine methylation status of MGMT Furthermore, as well as sample application and accuracy,
optimum pH, temperature and thermal stabilities of the
gene in 20 samples of planocelular cancer of lip vermilion. proposed systems were also investigated.
For that reason, we performed methylation specific PCR
(MSP) based on amplification of bisulfite modified DNA
P108
with pair of primers specific for methylated and
unmethylated DNA in the promoter region of this gene. AMPEROMETRIC BIOSENSOR FOR
Hypermethylation of CpG islands in promoter region of HYPOXANTHINE DETECTION
MGMT was found in 4 out of 20 DNA samples from oral Dilek ODACI, Suna TĠMUR, Mustafa SEZGĠNTÜRK,
cancer patients (20%). Our results suggest that Erol AKYILMAZ, Erhan DĠNÇKAYA, Azmi
TELEFONCU
transcriptional inactivation of MGMT by hypermethylation
Ege University, Faculty of Science, Biochemistry
may participate in the pathogenesis of planocellular head
Department, 35100-Bornova, Izmir/TURKIYE
and neck carcinoma. timur@sci.ege.edu.tr
P107 Quality control is of utmost importance in food industry.
THICK FILM SENSORS BASED ON The establishment of a rapid and accurate method for the
LACCASE FROM Trametes versicolor determination of fish freshness is a requirement in the
marine food industry. Development of an efficient and
IMMOBILIZED IN POLYANILINE MATRIX cheap sensor to monitor the quality of fish is therefore, a
Suna TĠMUR1, Nurdan PAZARLIOĞLU1, Roberto desired goal. The continuing development and application
PILLOTON2, Azmi TELEFONCU1 of analytical methods are proceeding at a rapid pace and
1
Ege University, Faculty of Science, Biochemistry many methods have been proposed for the determination of
Department, 35100-Bornova, Izmir/TURKEY trace amounts of hypoxanthine (HX) which is a major
2
ENEA C.R. Casaccia, Via Anguillarese 301-I-00060- metabolite in the degradation of adenine nucleotide, and
Santa Maria di Galeria-Rome/ITALY accumulates in fish and meet continuously after death.
timur@sci.ege.edu.tr Autodegredation of adenosine triphosphate (ATP) in fish
tissue follows the pathway;
During the past two decades, bioelectrochemistry has
received increased attention. Progress of ATP→ADP→AMP→IMP→INO→HX
bioelectrochemistry has been integrated into analytical Where ADP is adenosine diphosphate, AMP is adenosine
applications, e.g. in biosensors working as detectors in monophosphate, IMP is inosine monophosphate, INO is
clinical and environmental analysis. The development of inosine and HX is hypoxanthine. Whereas IMP is one of
sensors, which are highly selective and easy to handle the major contributing factors to pleasant flavour of fresh
opens the door to the problem in analysis. On the other fish, its degradation product HX imparts the bitter ‗off-
hand, conducting polymers have enough scope for the taste‘. The level of hypoxanthine is generally used in the
development of various sensors. Sensor systems based on food industry as an index for evaluating meet or fish
conducting polymers also rely on sensible changes in the freshness.
optical and electrical futures of this kind of materials.
Turk J Biochem, 2003; 28(3), 62-224 133 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
In this study, an enzyme electrode based on xanthine further studies with different doses, different time interval,
oxidase (XO) was developed for the determination of HX. and more animal number are needed.
The HX biosensor employs the amperometric detection of
oxygen consumed by the enzymatic reaction catalyzed by P110
XO immobilizing on the oxygen electrode. The system was
PLASMA PHOSPHOLIPASE A2(PLA2) AND
calibrated for both hypoxanthine and xanthine,
respectively. Furthermore, as well as sample application ACETYLCHOLINESTERASE(AChE)
and accuracy, optimum pH, temperature and thermal ACTIVITIES IN TYPE 2 DIABETES
stabilities of the proposed system were also investigated. MELLITUS
Mustafa TEKE1, Suna TĠMUR1, Erdal DUMAN2, Figen
P109 ZĠHNĠOĞLU1, Candeğer.YILMAZ2, Azmi TELEFONCU1
1
THE EFFECTS OF GINKGO BILOBA Ege Univ., Faculty of Science, Biochemistry Department,
35100, Bornova/ĠZMĠR-TÜRKĠYE
EXRACT ON TISSUE ADENOSINE 2
DEAMINASE, XANTHINE OXIDASE, Ege Univ., Faculty of Medicine,Endocrinology and
Metabolism Diseases Division, Bornova/ĠZMĠR-TÜRKĠYE
MYELOPEROXIDASE AND
MALONDIALDEHYDE, NITRIC OXIDE mustafateke@mail.ege.edu.tr
LEVELS IN CISPLATIN-INDUCED One propose that type 2 diabetes mellitus is due to damage
NEPHROTOXICITY to neurons in the ventromedial hypothalamus or to a defect
in the action or properties of insulin or insulin receptors in
Mukaddes GÜLEÇ1, Mustafa IRAZ2, Ramazan YILMAZ3,
brain. Phospholipase A2(PLA2; EC 3.1.1.4) is a lypolitic
Hüseyin ÖZYURT4, Ömer AKYOL1, Ġsmail TEMEL1
enzyme that catalyses, the hydrolysis of membrane
Departments of 1Biochemistry and 2Pharmacology, Inonu phospholipids into the corresponding lysophospholipid and
University Medical Faculty, Malatya, 3Department of fatty acid, mainly arachidonic acid(AA). Arachidonic acid,
Medical Biology and Genetics, Suleyman Demirel which is a precursor of eicosanoids, prostaglandins,
University Medical Faculty, Isparta, 4Department of prostacyclins, tromboxanes and leukotriens, enhances the
Biochemistry, Gaziosmanpasa University Medical Faculty, glucose uptake and glucose in turn augments
Tokat acetylcholine(ACh) release. Acetylcholinesterase(AChE;
mukaddesgul@hotmail.com EC 3.1.1.7) plays a key role in cholinergic transmission by
This study was carried out to determine if Ginkgo Biloba catalysing the rapid hydrolysis of the neurotransmitter ACh
Exract (GBE) exerts a beneficial effect against cisplatin- into acetate and choline. Recent studies in humans
induced renal failure in rats. Sprague Dawley rats were indicated that the cholinergic effects of ACh on insulin
divided into four groups and treated as follows: 1)control, secretion are mediated through muscarinic receptors,
untreated rats (n=7); 2) rats treated with i.p. injection in a located on the beta cell plasma membrane. To date both
single dose of 7 mg/kg body wt CDDP (Cisplatin, Ebewe) enzymes were thought to be differentiated in diabetic
(n=8); 3) rats treated with CDDP plus i.p. injection of 10 patients in various conditions.
mg/kg body wt vit E (Evigen-Aksu, Turkey) (n=9); 4) rats The present study was undertaken to emphasize the
treated with CDDP plus oral administration of GBE in the relationship between type 2 diabetes and plasma PLA2 and
dose of 100 mg/kg body wt (n=7). AChE activities. Venous blood samples were taken from
CDDP was found to lead statistically significant increases all volunteers which are female and closer age into tubes
in plasma BUN and creatinine levels as well as urine micro containing EDTA. Healthy and type 2 diabetic patients
total protein (MTP) levels leading ARF in rats. Renal were grouped according to the routine biochemical
xanthine oxidase (XO) activities increased in all of groups. analysis, glucose tolerance test and antropometric
The increase of XO in CDDP+GBE-treated rats was characteristics. Insulin sensitivity was also conducted by
HOMA. Group 1-4 are; controls without family history of
statistically significant according to control (p 0.05) between MEGX levels and values of
seedlings with SA alone resulted in decreased level of
routine liver functional tests (prothrombin time, albumin,
chlorophyll (Chl), A and transpiration. Pq treatment led to
total bilirubin, alkaline phosphatase, aspartate
a decrease in Chl and protein contents and to a very strong
aminotransferase, alanine aminotransferase). The MEGX
inhibition of A. Pq-treatment did not affect the activity of
test is an index in evaluating hepatic function and it is also
RuBPC but highly increased the activity of the
quick and easy to perform and capable of determining
photorespiratory enzymes. Pre-treatment of seedlings with
residual liver function. This test should not be used for
SA fully blocked the inhibitory effect of Pq on A and
Turk J Biochem, 2003; 28(3), 62-224 146 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
preoperative assessment in patients with benign hepatic believe that electromagnetic fields have an important effect
tumors. on blood levels and require care in daily use.
CONTROL GROUPS EXPOSED GROUPS
P143 Day 50 Days 100 Day 50 Day 100
Mean±SD Mean±SD Mean±SD Mean±SD
ALTERATIONS OF WHOLE BLOOD (n=12) (n=12) (n=12) (n=12)
RESULTS IN RATS THAT HAVE BEEN WBC 3.78 ±2.98 3,02 ±2,01 WBC 3,15±2,96 2,49 ±1,31
EXPOSED TO LOW FREQUENCY NEU 0.40±0.23 0,58 ±0,44 NEU 0.40±0.33 0,42 ±0,1
MAGNETIC FIELDS (50 HZ) LYM 2.83±2,98 1,56 ±1,32 LYM 1.56±0.8 1,54 ±1,03
MONO 0.24±0.22 0,43 ±0,48 MONO 0.12±0.09 0,19 ±0,1
Beran YOKUS1, Dilek Ulker CAKIR2, Zülküf AKDAG3, EOS 0.13±0.08 0,34 ±0,34 EOS 0.06±0.03 0,27 ±0,33
Nuriye METE2, Cemil SERT 4 BASO 0.09±0.07 0,13 ±0,13 BASO 0.09±0.12 0,05 ±0,04
1 RBC 7.77±0.27 7,33 ±1,27 RBC 6.76±1.8 7,58 ±0,51
Dicle University, Veterinary Faculty Department of
HGB 14.9±0.97 14,04 ±1,16 HGB 13.45±1,94* 14,1 ±0,94
Biochemistry, Diyarbakir
HCT 67.69±2.42 63 ±10,9 HCT 59.08±15,4 64,6 ±4,38
2
Dicle University, Medical Faculty Departments of MCV 86,8±1,86 85,9 ±2,02 MCV 87.68±1,95 85,3 ±2,09
Biochemistry, and Biophysic3 Diyarbakir MCH 19.20±1.36 19,7 ±3,85 MCH 22.29±9,25 18,6 ±0,44
4 MCHC 21.73±1.91 22,98 ±4,39 MCHC 25.37±10.37 21,81 ±0,36
Harran University, Medical Faculty Departments of RDW 16.66±1.83 16,79 ±1,68 RDW 15.5±0.94 16,32 ±1,33
Biophysics, Urfa PLT 672,75±327,4 503,3 ±247,1 PLT 703.27±289,4 525,3 ±217
MPV 10,04±0.89 8,44 ±1,12 MPV 8.43±0.81* 9,02 ±1,18
Exposure to a low frequency electromagnetic field (EMF)
PCT 0,77 ±0,23 0,5 ±0,15 PCT 0,65 ±0,21 0,51 ±0,09
(50 Hz) has some risks for health. One of the interaction PDW 19,7 ±1,62 19,62 ±1,65 PDW 18,5 ±1,35 20 ±2,14
mechanisms of magnetic fields with biological systems is
As compared to control *p0.05) compared with controls.
immunoradiometric assay, The results were analysed using No significant difference (P>0.05) was observed between
non-parametric statistical methods. Estrogen-regulated cut- any of the apoB XbaI genotypes and serum lipid and
Turk J Biochem, 2003; 28(3), 62-224 153 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
lipoprotein parameters. Associations of apoE years, 4.Type 2 diabetics ; duration 10 years). The results
polymorphism with the lipids analyzed were consistent indicated that both GFAT activity and UDP-GlcNAc levels
with the well-identified effects of apoE: E4 significantly were significantly increase in Type 2 diabetes patients with
(P0.05) were Also less increase in the levels of other two groups were
found in serum apoAI, apoE and Lp(a) by apoE alleles. observed. Correlation between all data was evaluated
The E4 allele was associated with increased serum apoB statistically by means of other biochemical parameters.
(P0.01). TOBACCO) ON HEMATOLOGICAL
These results suggest that excitotoxic hippocampal injury PARAMETERS
induced by convulsant doses of KA leads to oxidative Metin Kılınç*, Erdoğan Okur**, Fatma Ġnanç*, Ergül
stress in mitochondria and, the up-regulation of Trx may
Belge KurutaĢ*, Ġlhami Yıldırım**
be related its ROS scavenging function during this process.
Kahramanmaras Sutcuimam University, Faculty of
Medicine, Departments of Biochemistry* and
P174 Otolaryngology**. 46050 Kahramanmaras/TURKEY.
HIGH GLUCOSE EFFECT ON mkilinc@ksu.edu.tr
GELATINASES SECRETION BY Purpose: Nicotine is used in different forms including
ENDOTHELIAL CELLS smokeless tobacco. A special kind of smokeless tobacco
Manuela NICOLAE, Magdalena TIRCOL, Dorin also known as Maras Powder (MP) is widely used in South
ALEXANDRU eastern region especially Kahramanmaras, Gaziantep and
Institute of Cellular Biology and Pathology other south eastern cities of Turkey. It is obtained from a
“N.Simionescu”, tobacco species, Nicotina rustica L. (NRL) and ash of oak
or grapevine wood. The aim was to investigate the effect of
Laboratory of Extracellular Matrix, 79691,Bucharest/ nicotine on haematological parameters in MP users.
ROMANIA
Method: Sixty-nine MP users from Kahramanmaras and its
dalexandru@simionescu.instcellbiopath.ro
environs and 30 healthy controls who did not use MP were
Angiogenesis is one of the complications that appear in included in the study. We measured Soluble transferrin
diseases like diabetes. Two members of MMP family, reseptor (sTfR), transferrin (tf), ferritin, iron, iron binding
MMP-2 and MMP-9 play an important role in capacity (TIBC), white blood cell (WBC), neutrophil,
neovascularization process. Recent studies have revealed lymphocyte, monocyte, eosinophil, basophil, hemoglobin
that some drugs (acetylsalicylic acid, captopril, statins), (hgb), hematocrit (hct), MCV, MCH, MCHC, RDW,
that are not specific inhibitors of MMPs, are potent Platelet levels in the blood samples of MP users and
inhibitors of angiogenesis. The aim of this study was to controls.
determine the influence of high glucose condition on the
Turk J Biochem, 2003; 28(3), 62-224 157 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
Results: Our results showed that while iron and WBC SUPEROXIDE DISMUTASE, CATALASE
levels were higher in MP users than the controls (p0.05). KILINÇ1, , ġermin GÜL2
1
Conclusion: Increased leukocyte counts in MP users may KahramanmaraĢ Sütçü Ġmam University, Medical Faculty,
be an indicator of the present inflammatory events in Department of Biochemistry, KahramanmaraĢ, TURKEY
2
various tissues. So, we assume that MP, because of either Çukurova University, Art&Science Faculty, Department
high nicotine content or high tobacco-specific nitroso of Chemistry, Adana, TURKEY
amines levels (TSNA), causes chronic inflammatory The analytical, intra-individual and inter-individual
changes in various cells, organs and systemic circulation. variations were determined for urine superoxide dismutase,
Keywords: Hematological parameters Maras Powder, catalase and also malondialdehyde, and the reference
smokeless tobacco. values were established. A total of 143 apparently healthy
people, 70 male and 73 female, were randomly selected
from villages and cities of the southern part of Turkey. No
P176 significant difference was observed between the male and
LIPID PEROXIDATION, ANTIOXIDANT the female subjects. The mean (standard deviation) of the
DEFENCE SYSTEM AND ACID-BASE population investigated for superoxide dismutase was 5.36
(2.72) U/mg protein, for catalase was 0.52 (0.33) U/mg
STATUS IN PLACENTAL TISSUE protein, and also for malondialdehyde was 0.27 (0.12)
ACCORDING TO THE ROUTE OF nmol/mg protein, respectively. The analytical, intra-
DELIVERY individual and inter individual variations were assessed in
1
Fatma INANC, 2Alanur GUVEN, 1Metin KILINC, 1Ergul 15 apparently healthy subjects and were found to be;
BELGE KURUTAS, 3Gurkan CIKIM, 2Hakan KIRAN superoxide dismutase: 4.0%, 8.6%, and 31.0%, catalase:
3.5%, 15.0% and 29.5%, malondialdehyde 7.2%, 54.0%
During labour it is known that increased oxidative stress and 14.9%, respectively. The results of the index of
disturbs the balance between the oxidant-antioxidant individuality showed that reference values of
systems. Even though there are various publications that malondialdehyde could be used for diagnostic purposes
labour has an increasing effect on oxidative stress the except superoxide dismutase and catalase.
information about the effect of the mode of delivery on
oxidant and antioxidant systems is not decisive and
sufficient yet. P178
The aim of this study was to find out the degree of BIOLOGICAL CHARACTERIZATION OF
oxidative stress which the newborn is exposed to during CELL LINES ESTABLISHED FROM MC29
delivery and to investigate the state of the antioxidant
system and to see whether this showed any changes VIRUS-INDUCED TRANSPLANTABLE
according to the mode of labour. HEPATOMA IN CHICKEN
This study included 36 elective cesarean section and 37 Radostina ALEXANDROVA1, Ivaylo ALEXANDROV1,
normal vaginal deliveries. All of the patients had normal Evelina SHIKOVA1, Ivan IVANOV1, Plamena.
singleton pregnancies between 37 and 42 weeks gestation. JORDANOVA1, Weselina TSENOVA2, Martina
Immediately after delivery a segment of umbilical cord POTURNAJOVA3 Kristina HLUBINOVA3, Andrea
was double clamped and blood was drawn from both the PASTORAKOVA3, Veronika ALTANEROVA3, Cestmir
umbilical artery and umbilical vein into separate 5-ml pre- ALTANER3
1
heparinized plastic syringes. The blood samples were Institute of Experimental Pathology and Parasitology,
analyzed within 5-15 minutes of collection on a blood gas Bulg. Acad. Sci., Acad. G. Bonchev Str., Bl. 25, Sofia 1113,
analyzer for pH, carbon dioxide (pCO2), oxygen (pO2), Bulgaria, 2Queen Ioanna Hospital, 8 Bialo More Str., Sofia
bicarbonate, oxygen saturation and base excess. Placental 1527, Bulgaria, 3Cancer Research Institute, SAS,
samples were collected immediately on ice, washed with Bratislava, Slovakia
cold 0,9 per cent NaCl and stored at -20ºC. The placental
rialexandrova@hotmail.com
lipid peroxidation levels, superoxide dismutase (SOD) and
catalase (CAT) enzyme activities were evaluated The permanent cell line LSCC-SF-Mc29 was established
spectrophotometrically. from a transplantable chicken hepatoma induced by the
myelocytomatosis virus Mc29. The cell line was cloned
The levels of lipid peroxidation and the activities of CAT
and subcloned and four sublines were isolated: E7, E10,
and SOD increased in the plasental samples of cesarean
G9B4 and D6E10. The aim of the present study was to
section compared to normal vaginal deliveries (p LSCC- Sham controls received similar treatment except for
SF(Mc29) > D6E10 > E10 > G9B4. The most tumorigenic occlusion of the carotid blood flow. ROS were determined
were E7 cells: 85 – 100 % of the inoculated chickens by the CL technique in the fresh brain tissue samples.
developed tumours at the site of injection after 4-14 days Specimens were put into vials containing PBS-HEPES
latent period. G9B4 cells exhibited the lowest tumorigenic buffer. ROS were quantitated after the addition of luminol
potential – tumour growth appeared in only 12-25 % of (quantitates H2O2, OH-, HClO) and lucigenin (selective for
implanted chickens. While many organs were examined O2-) for a final concentration of 0.2 Mm. Counts were
tumour nodules were observed only in the liver (3 cases) obtained at 15 sec. intervals and the results were given as
and pancreas (1 case) of four chickens implanted with E7 the area under curve (AUC) for a counting period of 5 min
cells. E7 cells were also found to induce tumours in 5-6 and corrected for wet tissue weight (rlu/mg tissue).
weeks old nude mice when administered subcutaneously at
doses of 5, 7.5, 10 and 20 x 106 cells/mouse. The electron- Both luminol and lucigenin CL counts were significantly
microscopic investigations showed that the cells from all higher in all groups when compared with the healthy
lines were virus-producing. Liver and kidney tumours were controls. For the diabetic+I/R samples lucigenin CL
observed after intravenous inoculation of cell-free culture measurements were significantly increased with respect to
fluid from the cell cultures in 1-day old 15I White Leghorn both diabetic (p0.05, p>0.05). It is clear that, the oxidative injury in
OXIDATIVE INJURY IN CEREBRAL cerebral ischemia reperfusion becomes intensified with
ISCHEMIA REPERFUSION EXPOSED TO diabetes through excessive ROS generation.
DIABETIC RATS
Pınar ATUKEREN(1), M.Koray GUMUSTAS(1), Meral P180
YUKSEL(2), Ugur AKSU(3), A.Suha YALCĠN(2)
(1)
SINCREASED LEVELS OF NITRIC OXIDE
Istanbul University, Cerrahpasa Faculty of Medicine, DERIATIVES IN INDUCED SPUTUM IN
Dept. of Biochemistry, (3)Faculty of Sciences, Dept. of
Biology, (2)Marmara University, School of Medicine, Dept.
PATIENTS WITH CHRONIC
of Biochemistry,Istanbul, TURKEY OBSTRUCTIVE LUNG DISEASE (COPD)
p_atukeren@yahoo.com Aysun Bay Karabulut1, Süleyman SavaĢ Hacıevliyagil2,
Levent Cem Mutlu2, Ġsmail Temel1, Hakan Günen2 ,
Reactive oxygen species (ROS) are believed to be involved
Muzaffer Koçer1
in the pathogenesis of a variety of central nervous system
disorders, including cerebral ischemia-reperfusion (I/R) Inonu University, Faculty of Medicine, Department of
injury. During cerebral ischemia a number of events may Biochemistry1 and Pulmonary Diseases 2, Malatya, Turkey.
occur that predispose the brain to the formation of oxygen e.mail: abkarabulut@inonu.edu.tr,
free radicals. After reperfusion, these events can set off a sshacievliyagil@inonu.edu.tr
cascade of other biochemical and molecular sequale, such
as the xanthine/ xantine oxidase reaction and Chronic Obstructive Lung Disease (COPD) is a major
phospholipase activation, leading to free radical cause of chronic morbidity and mortality throughout the
production, especially superoxide anion (O2-), and causing world. One of the prevalent theory concerning of
additional central nervous system damage. On the other pathogenesis of COPD is that nitric oxide (NO) plays an
hand diabetes accelerates maturation of neuronal damage, important role as an inflammatory mediator in the airways.
increases infarct volume, and induces postischemic In this study, sputum induction was performed in 25
seizures. patients with COPD and 13 normal control subjects. Level
The aim of the present study was to investigate the of NO was measured in sputum samples. Total nitrite
oxidative damage in diabetic rats exposed to cerebral I/R levels in induced sputum were significantly higher in
injury by measuring chemilumiscence (CL). Male Wistar patients with COPD than in normal controls (341,5 164,8
Albino rats were divided into 4 groups as : control, mmol/L vs 95,028,2 mmol/L, p0.05).
and Mg2+ stress conditions. Absence of Ca2+, Mg2+ and Cardiovascular events were noted in 21 patient.
Zn2+ caused higher values of SOD and CAT in all organs Hypertension prevalence and CRP concentrations were
of M. pulegium than control and maximum increases were also higher in ESRD patients with CVD. All these
obtained in roots as 213.6 4.2 and 45.5 1.3 IU/mg; abnormalities in lipid/apolipoprotein status, elevated serum
139.9 2.7 and 29.2 0.5 IU/mg; 140.4 3.0 and 28.0 CRP concentrations, and hypertension may act
0.6 IU/mg in the absence of Ca2+, Mg2+ and Zn2+, synergistically with smoking, hypercoagulability and other
respectively. Whereas the activities increased above classical cardiovascular risk factors and contribute to the
control levels under excess Ca2+ and Mg2+ stress cardiovascular events in ESRD patients.
conditions, the values were lower than control under excess P188
Zn2+ conditions. Whereas AsA-dep POD activities in Ca2+,
Mg2+ and Zn2+ stress conditions were usually lower than DIFFERENTIATION OF K562 CELL LINE
control, the lower Gua-dep POD activity values were UNDER THE EFFECT OF HIGH
obtained only in leaves. All these antioxidant enzyme EXTRACELLULAR MAGNESIUM AND
activities correlated positively with increasing Cu2+ EXTREMELY LOW FREQUENCY
concentrations in all M. pulegium organs. SOD and CAT ELECTROMAGNETIC FIELDS
Turk J Biochem, 2003; 28(3), 62-224 162 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
1
G.AyĢe ĠNHAN , ġule ÖNCÜL1.M.Tunaya KALKAN2 the isolate GMBAE 42 have been submitted to NCBI,
1
Marmara Univ.School of Medicine,Biophysiscs NLM, NIH and GenBank nucleotide sequence databases
Dept.,34716,Ġstanbul,Turkey. under the accession number AY152839. Enzyme
2 production carried out by shaking flask cultivation of the
Ġstanbul Univ.Cerrahpasa Faculty of strain at 37°C and pH 10.5 in protein rich medium. The
Medicine,Biophysiscs Dept.,Ġstanbul,Turkey.
highest alkaline protease activity was observed at the late
aysein9@yahoo.com stationary phase of cell cultivation. The extra-cellular
Magnesium, the second most abundant cation in the alkaline protease in culture filtrate was highly purified by
intracellular environment is involved in a large variety of DEAE-cellulose anion exchange chromatography followed
metabolic functions. In the last years researches have by ammonium sulfate precipitation step. 57% of enzyme
shown that magnesium is also involved in regulation of cell activity available in the culture filtrate was obtained by 16-
proliferation and differentiation. The erythroleukemia ( fold purification. The molecular weight of enzyme was
chronic myeloid leukemia) cell line K562, was induced to found to be 64 kDa SDS-PAGE analysis. Michaelis-
differentiate with hemin. In order to evaluate the effect of Menten constant (Km) and turnover number (kcat) of
high extracellular magnesium on differentiation cells were enzyme was estimated as 1.8 mg ml-1 Hammarsten casein
kept above physiological levels ranging from 0.75mM to and 14.47 min-1 (specific activity: 4628 U mg-1, protein
2.00mM. Cells were then stained with trypan blue and concentration: 0.144 mg ml-1), respectively. Optimum
counted on the fourth day of induction with temperature of enzyme was found to be 60°C, however it is
hemacytometer. In contrast to the results obtained with shifted to 70°C after addition of Ca2+ ions in 5 mM
promyelocytic leukemia HL-60 cell line an increase in concentration. The enzyme was stable between 30-40°C
differentiation (%10-%40) and also a moderate increase in intervals when incubated for 2 hrs at pH 10.5. Only 14%
proliferation (%10) were observed. However these results activity loss was observed at 50°C at the same incubation
imply that magnesium is able to change the differentiation time and pH. Optimal pH of the enzyme was found to be
pattern. The cells were also exposed to extremely low 11.3. Enzyme did not show any activity loss at pH values
frequency electromagnetic field (ELF-EMF), 50 Hz, 5mT, between 9.0 to 12.2 when incubated for 24 hours at 30°C.
in similar extracellular magnesium concentrations at 38 and 76% activity losses were observed at pH values
different time sequences. One hour exposure at the time of 12.7 and 13.0 respectively at the same incubation time and
hemin induction caused a decrease in differentiation on the temperature. The activation energy of the Hammarsten
other hand when the cells were exposed each day for one casein hydrolysis by purified enzyme was found to be
hour an increase in differentiation was observed. These 10.59 kcal mol-1. The treatment of enzyme by active site
results demonstrate that the impact of ELF-EMF on living inhibitors iodoacetate, ethylacetimidate, phenylglyoxal,
systems depends on the exposure time sequence, which iodoacetimidate, n-ethylmaleimidate, n-bromosuccinate,
reflects the non-linear character of the interaction, and diethylpyrocarbonate, n-ethyl-5-phenyl-iso-xazolium-3'-
imply that this is affected by extracellular magnesium sulfonate did not affected the enzyme activity. The strong
concentration. inhibition of enzyme by phenylmethanesulfonyl-fluoride
(PMSF) treatment suggested that enzyme is a serine
P189 alkaline protease. Enzyme was stable in the presence of the
PURIFICATION AND 1% concentration of Tween-20, 40, 60, 80 and 0.2% SDS
after 1 hour incubation at 30°C and pH 10.5. Only 10%
CHARACTERISATION OF ALKALINE activity loss was observed by 1% sodium perborate (SPB)
PROTEASE FROM NEWLY ISOLATED at the same incubation conditions.
Bacillus clausii GMBAE 42 Key Words: Alkaline protease; Bacillus clausii; enzyme
Dilek Kazan1,2, A. Akın Denizci1, Mine N. Kerimak Öner3, purification; kinetic properties, active site inhibitors,
Altan Erarslan1,4 enzyme stability.
1The Scientific and Technical Research Council of Turkey
(TUBITAK), Research Institute for Genetic Engineering P190
and Biotechnology (RIGEB), Marmara Research Center DETERMINATION OF CHROMIUM(VI) BY
Campus (MRC), P.O. Box 21, 41470 Gebze – Kocaeli / A CATALYTIC SPECTROPHOTOMETRIC
TURKEY
2
METHOD IN THE PRESENCE OF p-
Marmara University, Faculty of Engineering, Department AMINOBENZOIC ACID
of Chemical Engineering, Göztepe Campus, 81040
Ziverbey-Kadıköy, Ġstanbul / TURKEY Angelina STOYANOVA
3
Kocaeli University, Köseköy Technical Collage, Division Higher Medical Institute-Pleven, Department of Chemistry
of Fermentation, 41135 Ġzmit – Kocaeli /TURKEY and Biochemistry & Physics and Biophysics,
4
Kocaeli University, Faculty of Arts and Sciences, 5800 Pleven/BULGARIA
Department of Chemistry, Division of Biochemistry, 41300 astoy@abv.bg
Ġzmit-Kocaeli / TURKEY Chromium(VI) is a strong oxidizing agent and possesses
kazan@rigeb.gov.tr high toxicity to humans and animals due to its carcinogenic
An extra-cellular alkaline protease producer Bacillus strain and mutagenic properties. That is why the determination of
capable of growing under highly alkaline conditions was chromium in environmental and biological samples is of
isolated from compost. Strain was identified as Bacillus great interest.
clausii according to the investigations on the physiological
In this work a catalytic spectrophotometric method for the
properties, cellular fatty acid composition, G + C content
of genomic DNA and 16SrRNA gene sequences analysis determination of chromium(VI) is proposed. The method is
and designated as GMBAE 42. 16S rRNA sequence data of based on the catalytic effect of chromium(VI) on the
Turk J Biochem, 2003; 28(3), 62-224 163 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
oxidation of sulphanilic acid (SA) by hydrogen peroxide in
the presence of p-aminobenzoic acid (PABA) as an P192
activator.
The reaction was followed spectrophotometrically by
THE LEVELS OF LEPTIN IN TIP I
tracing the formation of the reaction product at 360 nm DIABETES MELLITUS AND OBESITY
after 15 minutes of mixing the reagents. Berna ASLAN,Alev KURAL,ġebnem Ciğerli,Nezaket
On the bases of the investigations made, the optimum EREN
reaction conditions were established: The Biochemistry and Clinical Biochemistry Laboratory of
4.0x10-3 mol l-1 SA, 0.57 mol l-1 H2O2 , 1x10-3 mol l-1 Sisli Etfal Education and Research Hospital
PABA and 0.04 mol l-1 acetic acid – boric acid - The prevalance of type 2 DM has shown a dramatical
orthophosphoric acid buffer solution (pH 6.6), at 50 oC. increase in the last 20 years. The increase in obesity,
The linear range of the calibration graph was up to 140 ng decrease in the physical activity level and the changes in
ml-1 and the detection limit was 10 ng ml-1. Interferences the feeding habits are thought to be responsible. Besides
of Cu(II) and Cr(III) ions were masked. The method was being a serious disorder obesity plays a significant role in
applied to the analysis of Cr(VI) in industrial water with the pathogenesis of other disorders. For those reasons we
recoveries of 95.2 - 104.3 % and a mean RSD (n=6) of examined a new hormon called leptin in a patient
5.6%. population including obesity and type 2 DM. 80 patients
consisting of obese and nonobese type 2 DM patients and
Keywords: chromium(VI), catalytic method, sulphanilic obese and nonobese nondiabetic patients are examined in
acid, p-aminobenzoic acid, industrial water our study. BMI is calculated in those patients. Using leptin,
blood sugar, HbA1c, lipid profile( total kol, HDL-Kol,
P191 LDL-Kol.), insuline values in hunger and HOMA-IR
formule, IR values are facind. The results of all parameters
MOLECULAR MECHANISMS OF are are coralated with leptin and groups.As a conclusion
INTERACTION BETWEEN C1Q we determined that serum leptin level varies according to
COMPLEMENT SUBCOMPONENT AND sex, it is higher in females than males, that it has positive
IMMUNOGLOBULINS/ corelation with BMI: We also determined that leptin which
has multifactoriel effects has no relation with type 2 DM
Michaela KOJOUHAROVA1 , Michaela GADJEVA1, but is a parameter dependent on BMI in obese patients.As a
Ivanka. TSACHEVA1, Alexandra. ZLATAROVA1, conclusion more studies must be achieved in order to
Ljubka. ROUMENINA1, Magdalena CHORBADJIEVA, clarify the effects and the interactions with other
Boris ATANASOV 2 , Uday KISHORE 3, 4 and Ken. B. molekules of that hormon which has been identifical
M. REID3 recently and thus new steps should be taken in the
1
Dept. Biochemistry, Sofia University, Bulgaria, 2Institute pathogenesis and treatment of obesity and accompanying
of Organic Chemistry, Bulgarian Academy of Sciences, BG diseases like DM.
3
MRC Immunochemistry Unit and 4Weatherall Institute of
Molecular Medicine, University of Oxford, UK
1
P193
mkojouharova@biofac.uni-sofia.bg
BIOCHEMICAL EFFECTS OF DIAZINON
C1q is a recognition subunit of the classical complement
cascade. The interaction of the globular C1q heads with
ON ANTIOXIDANT DEFENCE SYSTEM,
their ligands- IgG or IgM triggers the classical pathway. LIPID PEROXIDATION AND
Each globular head (gC1q) is composed of the C-terminal ACETYLCHOLINESTERASE ACTIVITY IN
parts of A-, B- and C- chain ( ghA,ghB and ghC). Recent DIGESTIVE GLAND OF Cyprinus carpio L.
evidence suggests that the gC1q region has a modular Elif ÖZCAN ORUÇ and Demet USTA
organisation and is composed of three, structurally and
functionally, independent modules which retain Cukurova University, Faculty of Arts and Sciences,
multivalency in the form of a heterotrimer. We have Department of Biology, 01330, Balcali- Adana/TURKEY
expressed ghA, ghB and ghC and their single-residue eozcan@cu.edu.tr
mutants (ghAR162A, ghAR162E; ghBR114A, ghBR114E,
We investigated the effects of diazinon, at different
ghBR114Q, ghBR129A, ghBR129E, ghBR163AghB163E,
ghBH117A, ghB117D, ghCR156A and ghCR156E in E. concentrations and exposure times in fish, Cyprinus carpio
coli as soluble fusion proteins linked to maltose-binding to elucidate the possible mechanism related to oxidative
protein. The functional activity of the wild types and stress as well as the inhibitory effect of diazinon on
mutants were examined by several kinds of ELISA assays. acetylcholinesterase activity. Cholinesterase inhibition is
Our obsarvations lead to the conclusion that the interaction considered a specific biomarker of exposure and effect for
of ghB and ghC with immunoglobulins have mainly organophosphorus pesticides. Biochemical studies were
electrostatic nature, whereas in ghA the hydropfobic recorded spectrophotometrically in fish exposed to 0.036
interactions are involved as well. Our results highlight the ppb, 0.18 ppb, 0.36 ppb sublethal concentrations for 5, 15,
importance of arginine and histidine residues within gC1q 30 days. Digestive gland was chosen because of its
domain in the interaction between C1q and IgG and IgM. important role in the first pass of biotransformation of
The main role of ghBArg114 was proved. lipophilic xenobiotics. After 5 days diazinon exposure,
Turk J Biochem, 2003; 28(3), 62-224 164 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
superoxide dismutase, glutathione peroxidase and catalase STREPTOZOTOCIN-INDUCED DIABETIC
enzyme activities decreased and malondialdehyde content RAT DIAPHRAGM
increased, while 15 and 30 days of treatment caused no
further changes in the parameters. Acetylcholinesterase AyĢe DEMĠRKAZIK*, IĢıl ÖCAL*, AyĢe DOĞAN**,
activity remained constant in all the treatment groups Abdullah TULĠ***, Ġsmail GÜNAY*
compared with controls. An induction of antioxidant * Cukurova University, Faculty of Medicine, Department
enzyme activity and malondiladehyde content, as observed of Biophysics, 01330, Adana/TURKEY
in 5 days of diazinon exposure, may represent a first
response in this study, followed by an adaptation of ** Cukurova University, Faculty of Medicine, Department
antioxidant system to pesticide exposure. Since of Physiology, 01330, Adana/TURKEY
malondialdehyde content increased after diazinon exposure *** Cukurova University, Faculty of Medicine,
it is thought that diazinon toxicity may be possitively Department of Biochemistry, 01330, Adana/TURKEY
correlated to oxidative stress. Results of this study also
e-mail: ademir@cu.edu.tr
indicate that diazinon exposure may not essentially alter
the acetylcholinesterase activity, but may enhance lipid Electromagnetic fields can modify molecular structure, and
peroxidation to fish digestive gland. they play an important role in diverse physilogical
processes. Evidence obtained indicates that
eloctromagnetic fields can influence man and a wide range
P194 of animals. The effects of acute and chronic magnetic field
on live organism have taken part in current researches.
THE EFFECTS OF RICH OF
MONOUNSATURATED OIL ACID Diabetes mellitus is metabolic disorder that is characterized
HAZELNUT OIL AND RICH OF mainly with high blood glucose concentration. On the other
hand the effects of alternating (AC) macnetic field on
POLIUNSATURATED OIL ACID FISH OIL skeletal mucsle biomechanic in diabetic patients have not
ON THE LIPID PROFILE OF been identified yet. With this in mind, we aimed to
HEMODİALYSIS PATIENTS. evaluate magnetic field could be used as therapeutic tools.
ġebnem CĠĞERLĠ,Nurcan ÖZYÜREK, Nezaket EREN, In the present study, totally 40 rats Wistar Albino weighing
Aysun TOKER 271±12 gr were used. They were divided into four groups;
The biochemistry and Clinical Biochemistry Laboratory of control (n=10), control group exposed to AC magnetic
Sisli Etfal Hastanesi field (n=10), diabetic groups(n=10) and diabetic group
exposed to AC magnetic field (n=10). 20 rats were in the
Cardiovascular disease is considered one of the most experimental group exposed to AC magnetic field, 10 of
important mortility reasons in end stage renal failure.One them (control) having magnetic field and the other ten
of the factors responsible for atherosclerosis related to were diabetic group. The rats in the experimental groups
üremia is hyperlipidemia.It is known that hyperlipidemia were exposed to AC Magnetic field with 5 mT force in 50
increases the risk of cardiovascular disease. Hz frequency during four weeks 3 hours a day. Then rats
In our research we have divided hiperlipidemia were anaesthesized, blood samples were taken from their
hemodialytic patients into 3 groups according to their lipid heart ventricule, and diaphragm muscle strips (0.053 ±.0,06
profiles.And then ın each group we divided then into g.) were taken from the rats in the each group.
subgroups according to usage of two different fish oil and That blood glocose concentration of the diabetic group
hazelnut oil we administered these oils at specific doses for exposed to AC magnetic field is compared with diabetic
2 months.Triglycerides , cholesterol, VLDL , HDL, LDL groups, plasma glucose level was significantly lower.
levels have been measured.At the end we didn‘t find Muscle twitch measured by isometric transducers was
statistically significant difference in all groups and observed through digital storage oscilloscope, and put into
subgroups for cholesterol ve VLDL between pre-treatment computer in order for analysis. Of the isometric twitch
of fish and hazelnut oil(P>0,05).However , LDL tensions (Ps), contraction time (CT), one-half relaxation
cholesterol and HDL cholesterol levels were statistically time (½RT) were determined.
significant betwen pre- and post treatment (P100μM for MDA-MB-453 cells. Results obtained showed Most of the drugs used today in the therapy of malignant
that ellagic acid could not be solely cancer-preventive diseases express their effects either through inducing or by
agent, but could also possess anticancer activity. These the direct production of reactive oxygen species (ROS).
findings point to the importance of elaborating in vivo Increase ROS generation causes additional engagement of
studies to further elucidate the antitumor action of ellagic
the antioxidant defense system (AODS) in the organism, as
acid.
Turk J Biochem, 2003; 28(3), 62-224 168 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
well as inducing changes in parameters indicating to the Group III- Normoalbuminuria 300mg/24 h (n=10) higher levels than m2 in K562 cells. The levels of m3 and
m2 mRNA's were compared at 1,3,5, 24 and 48 hours of
Group II- Microalbuminuria 30-300 mg/24 h (n=24) agonist challenge. When compared to the level of
Turk J Biochem, 2003; 28(3), 62-224 169 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
expression at one hour after carbachol treatment, a These decreases and increases were not significant
decrease in mRNA transcripts was observed for m2 and m3 istatistically. In alendronate treted group, alendronate did
receptors after five hours of challenge. not cause a sifnificant difference in the level of plasma
Acknowledgement: glutathione and lipid peroxide both on day 14 and 28 as
compared with saline treated group.
This work was supported by grant from L'ORĒAL
TURKEY (UNESCO)
P207
P206 ATHEROSCLEROTIC POLYMORPHISMS
IN POSTMENOPAUSAL WOMEN
THE EFFECT OF SYSTEMIC
WITH ESTABLISHED CORONARY
ADMINISTRATION OF ALENDRONATE ON
DISEASE
PLASMA GLUTATHIONE AND LIPID
PEROXıDE LEVELS FOLLOWING TOOTH Lale AFRASYAP1, Ibrahim BARIS2
1
EXTRACTİON IN RATS Mugla University, Health High School, Department of
1 2 1 Biochemistry, Mugla ;
Azize ġENER , Hatice ALTUNDAL , Bahar GÖKER , 2
Ertuğrul YURTSEVER1 Bogazici University, Department of Molecular Biology
1 and Genetic, Istanbul/ TURKEY
Marmara University, Faculty of Pharmacy, Department of
Biochemistry, 34668, ĠstanbuL/TURKEY laleafrasyap@hotmail.com
2 The incidence of coronary disease risk due to
Yeditepe University, Faculty of Dentistry, Department of
Oral Surgery, 34668, ĠstanbuL/TURKEY atherosclerosis is higher in men and postmenopausal
women than in premenopausal women. Although the
A wound can be described as the damage to a tissue
polymorphisms of the MTHFR (C677T and A1298C) and
integrity. The tissue damage can result from several
eNOS (G894T) genes were investigated in different
factors. One of the significant mechanism in cell damage is
population groups with coronary disease, very few studies
the destruction due to free radicals. Eventhough the
have addressed about the association between these
mechanism of the oxygen free radicals formation is
polymorphisms and coronary disease in postmenopausal
thoroughly underrstood, the role of these compounds on
women. The aim of study is to investigate if genetic
healing process in wounded tissue has not yet been clearly
mutations increase the risk of coronary disease in
elucitated.
postmenopausal women. The study was organized for 40
The aim of this study was to investigate the effect of the postmenopausal women with an intact uterus. They were
free radicals formation on plasma lipid peroxide and divided into two groups, according to angiography results.
glutathione levels during soft tissue healing following tooth 1- 25 women with >50% stenosis affecting at least one
exraction in rats. In addition, the effect of alendronate, artery were included in group with coronary heart disease
which is applied to prevent alveolar bone loss following (patients) 2-15 women with > 5.5 ms. The DF changes during induction are compared
up to 20–30 ms after the beginning of illumination. This
with simultaneously registered chlorophyll fluorescence
DF component forms mostly the first peak, I1, of the DF
transients and 820 nm absorption changes that correlate
induction curve. The time course of the concentration of
with P700 reduction. Both amplitudes and lifetimes of DF
components are modified typically during induction. It was centers in the Z+QA–QB= redox state shows that these
shown, that the first DF maximum, I1, appeared at 20–30 centers take part in the formation of both I1 and I2 (which is
ms after beginning of illumination, is produced by both observed after 100–150 ms of illumination), and are
sub- and millisecond DF components. It correlates with suggested to correspond to the millisecond component of
formation of high relative concentration of opened DF dark relaxation. The possible role of transmembrane
Photosystem II (PS II) reaction centers with secondary electrical gradient formation for the appearance of DF
quinone acceptor at reduced or semi-reduced state as well maximums is discussed.
as with transmembrane electrical gradient formation. The Acknowledgments. This work was financially supported by
second maximum, I2, is observed at 100–150 ms of the Swiss National Science Foundation (SCOPES 2000–
illumination and includes predominantly millisecond DF 2003 grant № 7BUPJ062408.00/1).
component. Its rise is associated with reaction center
reopening as a result of QB= reoxidation. At the end of the
first second of illumination a minor peak, I3, was registered P216
that is associated with slowest component of light emission
from closed PS II reaction centers. We are supposed that PROMPT AND DELAYED CHLOROPHYLL
the emission of observed DF components is a result of FLUORESCENCE OF INTACT LEAVES IN
charge recombination in PS II reaction centers at different THE PRESENCE OF PHOTOSYNTHETIC
redox states: Z+QA–QB– – for sub-millisecond, Z+QA–QB= – HERBICIDES
for millisecond and SiZQA–QB= – for slowest DF
component, respectively. Petar LAMBREV1, Vassilij GOLTSEV, Reto J.
STRASSER2
Acknowledgments. This work was financially supported by
the Swiss National Science Foundation (SCOPES 2000– Dept. Biophysics and Radiobiology, Faculty of Biology,
2003 grant № 7BUPJ062408.00/1). “St. Kliment Ohridski” University of Sofia,
8 “Dragan Tzankov” Blvd., 1164 Sofia, Bulgaria. E-mail:
P215 goltsev@biofac.uni-sofia.bg
A MATHEMATICAL MODEL OF THE 1
Institute of Biophysics, Bulgarian Academy of Sciences,
KINETIC COMPONENTS OF “Acad. G. Bonchev” Str., Bl.21, 1113 Sofia, Bulgaria. E-
MILLISECOND DARK DECAY OF mail: lambrev@spnet.net
DELAYED CHLOROPHYLL A 3
Bioenergetics Laboratory, University of Geneva, CH-
FLUORESCENCE IN LEAVES DURING THE 1254 Jussy – Genève Switzerland
FIRST SECOND OF INDUCTION Prompt fluorescence (PF) and delayed fluorescence (DF)
Petko CHERNEV, Vassilij GOLTSEV, Reto J. of chlorophyll a are extremely sensitive intrinsic probes for
STRASSER1 the function of the photosynthetic apparatus in vivo,
however, they are still not sufficiently understood. We
studied the effects of photosynthetic herbicides that block
Turk J Biochem, 2003; 28(3), 62-224 173 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
the Photosystem 2 electron transport on the PF and subjected to 2D-PAAG electrophoresis. Upon comparing
millisecond DF induction curves and the DF decay the protein pattern of secreted and ionically bound cell wall
kinetics. The herbicides diuron and atrazine were applied proteins we observed the appearance and accumulation of
in various concentrations to 14-days-old pea plants through certain specific proteins from salt adapted lines. Phage
their stems. PF and DF were measured simultaneously display has been used in order to generate specific
from detached leaves using a phosphoroscope fluorometer. antibodies against some proteins characteristic for NaCl-
High-resolution fluorescence induction transients (OJIP selected lines. Proteins were transferred to a nitrocellulose
curves) were registered using a HandyPEA fluorometer membrane after 2D PAAGE and protein spots were used
(Hansatech, UK). The presence of diuron or atrazine in the for the selection by phage display. Specific antibodies were
leaves had a profound effect on both luminescence types
selected after 5 round of panning from human synthetic
that could be measured quantitatively. The herbicides
single-chain Fv (scFv) phage display library (Griffin 1).
diminished the second peak in the fast phase of the DF
Futher investigations are in progress in order to elucidate
induction curve, I2, which is supposed to be related to the
intersystem electron transport. The first maximum, I1, was the possible use of these antibodies as markers for
less sensitive, in accordance with the concept that it arises adaptation to salt stress.
due to a transiently generated transmembrane electrical Research was supported by grant 3457 from the Science
gradient. The second component of the slow-phase DF Fund of Sofia University
peak, I5, was strongly inhibited, suggesting that it is a result
of a partial reopening of the reaction centres in non-treated P218
samples. Only I4, which is supposed to reflect the thylakoid
membrane energization, was expressed in the slow phase at INTEGRATING MUTATION DATA AND
high herbicide concentrations. In this case I4 might be STRUCTURAL ANALYSIS OF THE G6PD
related to a proton gradient built by Photosystem 1 via ENZYME
cyclic electron transport. PF was progressively quenched
by increasing the herbicide concentration even when the Erdinç A. YALIN, Kıymet AKSOY
electron transport was completely inhibited. This effect Çukurova University, Faculty of Medicine, Department of
could not be fully explained by the now-accepted Biochemistry, 01330, Adana/Turkey
quenching by oxidised plastoquinone.
eyalin@cukurova.edu.tr
Acknowledgments. This work was financially supported by
the Swiss National Science Foundation (SCOPES 2000– Glucose-6-phosphate dehydrogenase (G6PD, MIM#
2003 grant № 7BUPJ062408.00/1). 305900) is a cytosolic enzyme encoded by a house keeping
X-linked gene whose main function is to produce NADPH,
a key electron donor in the defence against oxidizing
P217 agents and in reductive biosynthetic reactions. Hereditary
GENERATION OF SPECIFIC ANTIBODIES deficiency of human G6PD is one of the most common
AGAINST STRESS-RELATED PROTEINS human enzyme deficiency. The deficiency affects an
estimated 400 million people worldwide with gene
FROM NaCL ADAPTED EMBRYOGENIC
frequencies ranging from 5% to 25%. G6PD deficiency is
CALLUS OF DACTYLIS GLOMERATA L. very prevalent in the Çukurova Region of Turkey, a gene
Mima I. PETKOVA, Magdalena I. CHORBADJIEVA and frequency about 8.2 % has been documented.
Mariela ODJAKOVA
Beside about 440 different G6PD variants have been
Sofia University, Faculty of Biology, Department of described based on biochemical and clinical characteristics,
Biochemistry, 1164 Sofia/Bulgaria over 125 distinct mutations of G6PD have been identified
modjakova@biofac.uni-sofia.bg to date. The relational databases integrates up-to-date
The physiological and biochemical changes in plant tissue mutations and structural data from various databanks.
in response to different types of osmotic stresses are not These databases and recently developed procedures
completely understood. Despite extensive research of provides insights into the molecular aspects and clinical
desiccation tolerance in plants, little is known about the significance of G6PD deficiency for researchers and
genes and their related proteins involved in stress-defense clinicians, and these web-based functions as a knowledge
mechanisms. The presence of some stress proteins at base relevant to the understanding of G6PD deficiency and
different stages of embryonic response shows that besides its management.
their protective function, they are important for More than 50% of the mutations in the G6PD gene have
embryogenic competence, too. been reported to be in severe (Class I) deficiency, and these
The aim of our research is to generate specific antibodies affect dimer interface and/or coenzyme binding cleft,
against stress-related proteins resulting in partial or complete loss of enzyme activity.
We studied the changes in the protein profiles of Among the 104 distinct mutations we analysed 53 (50.9 %)
intracellular, extracellular and ionically-bound cell wall mutations considered in Class I variants. We report here
proteins from embryogenic callus from Dactylis glomerata the results of systematic analysis of the effect of 53
L. genotype Embryogenic P, selected and maintained on mutations corresponding Class I variants, which can be
SH30 medium with different NaCl concentrations. Proteins explained in structural terms by their predicted effects on
from different cell compartments were isolated and protein stability.
Turk J Biochem, 2003; 28(3), 62-224 174 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
P219 P220
THE EFFECT OF CAFFEIC ACID THE ANTIULCEROGENIC EFFECT OF
PHENETHYL ESTER (CAPE) ON USNIC ACID ISOLATED FROM USNEA
CISPLATIN-INDUCED TOXICITY IN RAT LONGİSSİMA ON INDOMETHACINE-
LIVER TISSUES INDUCED GASTRIC ULCER IN RATS
Fehmi ODABASOGLU a,* Halis SULEYMAN b, , Ahmet
Elif ÖZEROL1, Mukaddes GÜLEÇ1, Ersin CAKIR c, Ali ASLAN d, Yasin BAYIR a, Mesut HALICI a,
FADILLIOĞLU2, Mustafa IRAZ3, Seda AĞLAMIġ3, Osman YUCELe, Cavit KAZAZ f and Erdal
Ömer AKYOL1 DEMIRCEYLANa
a
Inonu University, Faculty of Medicine, Departments of Ataturk University, Faculty of Pharmacy, Department of
1
Biochemistry,2 Physiology and 3Pharmacology, 44069 Biochemistry, 25240, Erzurum, Turkey.
Malatya, Turkey b
Ataturk University, Faculty of Medicine, Department of
eozerol@inonu.edu.tr Pharmacology, 25240, Erzurum, Turkey.
c
Ataturk University, Kazım Karabekir Education Faculty,
High doses of cisplatin have been known to produce
Dep. Chemistry, 25240, Erzurum, Turkey.
hepatotoxicity. However, little is known about d
pathophysiology of cisplatin-induced liver injury. The Ataturk University, Kazım Karabekir Education Faculty,
present study was designed to determine the effect of Dep. Biology, 25240, Erzurum, Turkey.
e
cisplatin on liver oxidant/antioxidant system and the Ataturk University, Sağlik Hizmetleri Meslek Yuksek
possible protective effect of caffeic acid phenethyl ester Okulu, 25240, Erzurum, Turkey.
(CAPE) on liver toxicity induced by cisplatin. Adult f
Ataturk University, Faculty of Science, Department of
female Wistar albino rats were divided into four groups Chemistry, 25240, Erzurum, Turkey.
(n=6 per group): Control, Cisplatin, CAPE, and fodabasoglu@yahoo.com
Cisplatin+CAPE. Cisplatin was injected intraperitoneally
(a single dose of 16 mg/kg bwt) to the second and the last In the present study, the antiulcerogenic effect of usnic
acid (UA) (a prototype of the dibenzofuran derivatives),
groups of rats. CAPE was applied to the rats with a dose of
isolated from diethyl ether extract of Usnea longissima on
10 mol/kg/day (i.p.) one day before and 5 consecutive indomethacine-induced gastric ulcers in rats was
days after cisplatin injection. At the 5th day of cisplatin investigated and compared with ranitidine. A total of 48
injection the experiment was finished and liver tissue was male, albino Wistar rats, weighing 180-190 g, have been
removed to study the activities of catalase (CAT), used for the experiments. UA and ranitidine were
superoxide dismutase (SOD), glutathione peroxidase administrated to the assigned groups of rats per orally, then
(GSH-PX), myeloperoxidase (MPO), xanthine oxidase the animals were sacrificed with high dose anaesthesia
(XO), adenosine deaminase (ADA), and levels of (thiopental sodium, 50 mg/kg). The stomachs of rats were
malondialdehyde (MDA) and nitric oxide (NO) in liver removed and the gastric damage (or ulcer) in the stomachs
tissue. was macroscopically evaluated. For the activity studies, 5,
10, 25, 50, 100 and 200 mg/kg doses of usnic acid were
The activities of SOD and GSH-Px were increased in tested. Antiulcer effect of UA were determined by
Cisplatin+CAPE and CAPE groups in comparison with comparing to the results obtained from ranitidine (150
Cisplatin groups. The activity of CAT was higher in mg/kg dose), used as positive control, and control groups.
Cisplatin+CAPE group than other three groups. The In general, gastric damage in the rat groups treated with
activity of XO was lower in Cisplatin group than control usnic acid and ranitidine was less than that of the control
group. Also, the activity of MPO was increased in groups. While the mean damage areas in rats receiving the
Cisplatin group in comparison with control and CAPE doses of 5, 10, 25, 50, 100 and 200 mg/Kg of usnic acid
groups. There were positive correlations between SOD and was 33.2±6.0, 22.9±5.3, 20.8±3.5, 16.7±11.7, 6.8±2.9,
MPO, SOD and MDA, NO and ADA in Cisplatin group. 18.2±8.6 mm2, respectively, they were 36.0±11.7 and
There were positive correlations between SOD and CAT, 6.0±2.1 mm2 in the control and ranitidine groups,
ADA and XO, and negative correlations between CAT and respectively. Among the treated doses, 50, 100 and 200
mg/kg doses of usnic acid showed potent antiulcerogenic
MDA, SOD and MDA in Cisplatin+CAPE groups. There
activity in comparison with control groups and also 100
was a positive correlation in CAPE group between NO and
mg/kg dose of usnic acid was most effective in preventing
CAT. of the gastric damage in rats. On the other hand, the
It can be concluded that CAPE prevents oxidative injury antiulcerogenic acitivity of 100 mg of usnic acid was
due to cisplatin in the liver tissue by increasing antioxidant roughly the same as ranitidine, and as statistically it was
enzyme activities and preventing MPO dependent reactive not significant (pA in intron 6, which can be detected by the organic chemicals including PAHs, PCBs, dioxins and
RPLF using DraI restriction enzyme. Studies have shown pesticides. Amoung these pollutants, PAHs, PCBs, dioxins
an association between DraI polymorphism and incidence induce one family of P450, P4501A in fish liver. P4501A
of lung, breast cancer and renal carcinoma. oxidatively metabolize these toxic compounds,
In this study, DNA was isolated from blood samples precarcinogen/ carcinogens to their epoxides and other
belonging to healthy Turkish individuals by oxygenated metabolites which in turn bind to DNA and
phenol:chloroform:isoamylalcohol extraction method. form DNA adducts leading to membrane impairment,
Qualitative analysis of DNA was performed by 0.4% cellular toxicity, mutation or even carcinogenesis. The
agarose gel electrophoresis and the concentration and induction of hepatic CYP1A and its monooxygenase
purity of the samples were determined activity 7-ethoxyresorufin O- deethylase (EROD) in fish by
spectrophotometrically by measuring the absorbance PAHs, PCBs and dioxins has been suggested as an early
values at 260 and 280 nm. To amplify the DraI restriction warning system, a ―most sensitive biochemical response‖
site on intron 6 of CYP2E1 gene, the optimized PCR
for assessing environmental contamination conditions. This
medium contained 10mM Tris-HCl, 50 mM KCl, 0.08%
Nonidet P40, 1.5 mM MgCl2, 0.2 mM dNTP mixture, 20 has implications for human fish consumption, as well as
pmol of each primer (5‘- for the health status of the organisms.
TCGTCAGTTCCTGAAAGCAGG-3‘ and 5‘- This study was carried out to determine if there exists a
GAGCTCTGATGCAAGTATCGCA-3‘), 0.4 U of Taq decrease in the concentrations of PAHs, PCB and dioxins
DNA polymerase and 200 ng of DNA. The cycles of PCR after the Great Canal Project in the Ġzmir Bay by
were optimized as follows: 94C, 3 min. for denaturation; measuring induction of cytochrome P4501A associated
61C, 1 min for annealing; 72C, 1 min for extension and a EROD activity and to compare these results with the
final extension at 72C for 6 min. The PCR product of previous studies. (Arınç and ġen, Marine Environmental
nearly 1000 base pairs was detected by 1.5% agarose gel Research.; 48, 147, 1999, Arınç, ġen and Bozcaarmutlu,
electrophoresis. For the detection of polymorphism, RFLP Pure Appl.Chem.; 72, 985, 2000.).In this study, two types
was performed on the PCR products. 20 µL of PCR
of fish species, Leaping Mullet (Liza saliens) as a pelagic
product was incubated with 4 U of restriction enzyme DraI
fish and Annular sea bream (Diplodus annularis) as a
at 37C for 18 hours and the results were visualized by
1.8% agarose gel electrophoresis. benthopelagic fish were examined for cytochrome P4501A
Turk J Biochem, 2003; 28(3), 62-224 179 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
associated 7- ethoxyresorofin O-deethylase (EROD) We have observed that serum GGT activities were high in
activity. Fish were captured on November 2002 from smokers (p 0.05)
microsomes were determined spectrofluorometrically. and also serum AST levels were higher in alcohol drinkers
Mullet caught from outer site of Bay had very low EROD than those in control subjects (p 0.05), nucleic acid content were exponentially degraded to the
whereas we have observed that saliva TSA levels were increase of the exposure time. This means that self-
higher in alcohol drinkers than those in controls (p 0.05) but urine TSA levels were molecular level.
much higher in alcohol drinkers than those in healthy So, low radiation doses, may influence cell biochemistry
subjects (p100nm) were isolated by
thiocarbamoyl group) was more active than B-3. ultracentrifugation at 4x106g.min. Since the particle
aggregation is a characteristic of hydrophobic nature of
P267 lipoproteins, as an index of aggregation, the turbidity
generated by vortexing and storage of chylomicrons was
HPYERLIPIDEMIA TREATMENT WITH measured spectrophotometrically at 680nm. In contrast to
ATORVASTATIN : HOMOCYSTEINE AND LDL, neither shaking nor prolonged storage at 4 oC
NITRIC OXIDE produced an increase in the optical density of chylomicron
solution indicating no aggregation took place. In a second
Çiğdem YENĠSEY1, Nihat ÖZGEL2, Mukadder SERTER1, series of experiment, ability of chylomicrons to interact
Zahit BOLAMAN2 with five different hydrophobic interaction
1 Adnan Menderes University, Faculty of Medicine, chromatography media (phenyl sepharose high
Department of Biochemistry and performance, phenyl sepharose 6 fast flow (low substance),
phenyl sepharose 6 fast flow (high substance), butyl
2 Department of Internal Medicine, 09100 Aydın/TURKEY,
sepharose 4 fast flow and octyl sepharose 4 flow. Typical
cyen2002@hotmail.com
elution profiles of chylomicrons through octyl, phenyl
OBJECTIVE: To determine the efficacy of atorvastatin to (high substance) and butyl sepharose columns showed two
mild hyperlipidemia, hypercholesterolemi, and peaks. Peak I material emerged with 4M NaCl in a
hyperhomocysteinemia. and to test the hypothesis that position corresponding to the void volume and peak II
endothelial NO elaboration is impaired in material eluted with water. Phenyl sepharose (high
hypercholesterolemi, and hyperhomocysteinemi. performance) media exhibited the maximum binding
METHODS: Patients (n=44) were randomly assigned to 6 strength towards chylomicrons among the five different
media.In the case of phenyl sepharose (low substance)
months of treatment with atorvastatin (10 mg/day) and also
column, an additional material was eluted with 3 M NaCl
control individuals (n=30) were selected according to their
Turk J Biochem, 2003; 28(3), 62-224 194 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
2
between peak I and II. These results indicate the Department of Medical Pharmacology, Experimental
heterogeneity of chylomicron surface hydrophobicity. Psychopharmacology Research Unit, Etlik 06018 Ankara –
Since the particle aggregation is a characteristics of Turkey
hydrophobicity of lipoproteins and the aggregation is
cbilgi@gata.edu.tr
believed to be the underlying cause of atherosclerosis,
fractionation of lipoproteins by hydrophobic interaction The aim of the present study was to investigate if the
chromatography may introduce a new approach into the chronic ethanol administration by liquid diet to rats may be
assessment of lipoprotein atherogeneicity. a dementia model.
Key Words: Aggregation, chylomicron, hydrophobicity, Female Wistar rats (188-244 g) were used in the study.
atherosclerosis, chromatography Ethanol was administered by a modified liquid diet with
4.8% (v/v) ethanol for 3 days followed by 25 days on a
P269 liquid diet in which the ethanol concentration was
increased to 7.2%. Control rats were pair fed with an
FRACTAL ANALYSIS IN MEDICAL isocaloric liquid diet not containing ethanol. Serum ChE
IMAGISTIC activity and blood ethanol concentration were measured at
M. Tudorie, D.E. Creanga, C. Murgulet* the end of the 4.8% ethanol consumption and after 35 days
of ethanol (7.2%) feeding and, just before, 24th and 72nd
Univ. Al. I. Cuza, Fac. of Physics, Biophysics and Medical
hours ethanol witdrawal period. Cognitive functions were
Physics, 11 A, Bd. Carol I,, Iasi-Romania, evaluated by step-down passive avoidance test system for
e-mail:dorina.creanga@email.ro
150 sec (cut-off time) in three individual groups of ethanol-
*University Hospital, Iasi, Romania administered, ethanol withdrawn (24th h withdrawal) and
Fractal analysis was carried out using adequate soft control rats. The data was evaluated by one-way analysis
packages for the calculation of fractal dimension for two of variance followed by Tukey‘s test for post-hoc
types of medical images: tomography films and electro comparison.
graphical recordings (variant of Kirlian images obtained in The daily ethanol consumption of the rats ranged from 11.5
the radiology clinic of the University Hospital and to 14.9 g/kg. ChE activity was found significantly
respectively in the Medical Physics Laboratory). The increased from 3rd day of ethanol (4.8%) consumption.
mathematical method utilized for the fractal dimension Serum ChE activities of the rats receiving ethanol (7.2%)
calculation was based on the ‗box-counting‘ algorithm. also increased significantly as compared to ethanol (4.8%)
The main type of tomographical image was the result of ingesting rats. Blood ethanol levels were measured as 200
brain investigation on the basis of a Siemens Computer and 2.2 mg/dl at 35th days of ethanol consumption (just
Tomograph; it was revealed that brain tumors leads to the before ethanol withdrawal) and 24th h of ethanol
increase of fractal dimension with about 10%. The electro withdrawal, respectively. Passive avoidance latency was
graphical images were obtained using an electrostatic found significantly reduced in the groups that just before
device designed and assembled by us, hands and feet of and 24th h of ethanol withdrawal as compared to control
normal subjects in comparison to pathological being rats.
studied by means of the fractal dimension. Statistical Our results suggest that serum ChE activity increased by
analysis of the differences between the values provided by
chronic ethanol consumption in rats and chronic ethanol
groups of normal and pathological cases showed
caused some marked impairments on the cognitive
significant results for brain tomography images as well as
functions Overall the data indicated that chronic ethanol
for electro graphical images of the hands but non-
feeding might be a model for evaluation cognitive
significant differences were revealed for feet images.
functions in rats.
References
1] B. Mandelbrot, The Fractal Geometry of Nature,
P271
Academic Press, 1975, New York
[2] G. Hartvigsen, The analysis of leaf Shape using Fractal AN OBSERVATION ON THE RAT
geometry, The American Biology Teacher, 62, (9), 2000, PNEUMOCARDIOGRAM WITH
663-665 NONLINEAR STRUCTURE
Tamer ZEREN1, Mustafa ÖZBEK2, Nuran EKERBĠÇER2,
P270 K.Gediz AKDENĠZ3
Celal Bayar University, Medical School, Departments of
A RAT DEMENTIA MODEL BY CHRONIC
Biophysics1 and Physiology 2, Manisa
ETHANOL CONSUMPTION AND
WITHDRAWAL: VALIDATION BY PASSIVE University of Ġstanbul, School of Science, Department of
Physics3, Ġstanbul
AVOIDANCE MEASUREMENT AND
SERUM CHOLINESTERASE LEVEL tzeren@bayar.edu.tr
1 2
Erdinç ÇAKIR , Turgay. ÇELĠK , Hakan KAYIR , 2 Introduction: The cardiorespiratory system function
Cumhur BĠLGĠ1, and I. Tayfun UZBAY2 requires the harmony between heart and lung: The volume
changes of these two organs is an evidence of vitality of
Gulhane Military Medical Academy, 1Department of organism and it has a very significant functional value for
Emergency Medicine, Laboratory of Biochemistry, respiratory gas exchange.
Turk J Biochem, 2003; 28(3), 62-224 195 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
Not only the volume changes of the lung but also the LDL-cholesterol (LDL) and HDL-Cholesterol (HDL)
cardiac actions can be recorded on the external airway: The levels in COPD patients and correlate their serum levels to
pneumocardiogram (PNCG) is a non-invasive record of the the severity of COPD.
pulsative air flow in the trachea coincident with hearth Fifty-two clinically stable male COPD outpatients (age
motions [1]. The PNCG signals was previously considered 62.4±6.9) with no concomitant disease were admitted to
to be useful for measurements of the respiratory mechanics the study. The patients were evaluated with clinical
and a model has been developed to explain dynamic findings, pulmonary function tests and arterial blood gas
respiratory impedance changes of external airway [2]. analyses and subgrouped according to the severity of
Material and Methods: Recently, the tracheal air flow COPD (FEV1; forced expiratory volume in one second, %
created by heart actions of the spontaneously breathing rats predicted). Serum Chol, TG, LDL and HDL levels were
could be obtained [3]. And, it has been emphasized that the measured by ILLab 1800 autoanalyser (ILLab test kits).
PNCG method may be useful for physiological studies of The statistical analyses were done by Pearson correlation
circulation system in the small laboratory animals: Briefly, coefficients and independent samples t-test. p50%) COPD
pneumocardiographic complex signals in Ref. [3]. It has patients were: Chol: 203±58.9, 218.6±54 mg/dl; TG:
been suggested that a non-linear model to be necessary for 126.8±100.8, 161.56±75.3 mg/dl; HDL: 43.1±13.2, 46±9.1
understanding of the fractals and dynamic behaviour of mg/dl; and LDL: 57.5±51.2, 109±69.3 mg/dl
PNCG [4]. In this presentation we propose a new nonlinear consequtively. Serum tryglyceride (p= 0.012 ) and LDL
model which may help to determine the reasons and/or (p=0.018). levels of severe COPD patients were lower than
importance of chaotic dynamic structure of mild-moderate patients A positive correlation was found
between serum cholesterol (p=0.001, r=0.473),
cardiorespiratory functions and which could let us to obtain
LDL(p=0.027, r=0.337) and severity of COPD.
simulate data.
[1] J.A Reitan and A. Lim; "Automated measurement and P273
frequency analysis of the pneumocardiogram", Anesth
Analg. Nov-Dec;57(6), 647-52 (1978) INTRACELLULAR TRAFFICKING OF PH-
[2] E. Bijaoui, P.F. Baconnier and J.H.T. Bates;
SENSITIVE LIPOSOMES
"Mechanical output impedance of the lung determined Roxana C. MUSTATA, Stefana M. PETRESCU
from cardiogenic oscillations", J Appl Physiol. 91: 859- Department of Molecular Glycobiology, Institute of
865 (2001) Biochemistry, Romanian Academy,
[3] M. Özbek, N. Ekerbiçer, M. Pehlivan, A. Akay, A. G. Spl. Independentei 296, 060031 Bucharest, Romania
Karakurt and T. Zeren; "Anestezi altında spontan solunum
roxanam@biochim.ro
yapan sıçanlarda pnömokardiografi", to be presented in
29th National Physiology Congress, Ankara-Turkey (01-05 Liposomes are used as drug delivery system, with the
September 2003) purpose of reducing substance toxicity and/or increase its
pharmacological efficacy. Although a number of liposome
[4] T. Zeren, M. Özbek, N. Ekerbiçer and K.G. Akdeniz;
formulations are already patented, not many data have been
"Pnömokardiogramda gözlenen düzensiz dinamik yapıların
reported on the intracellular trafficking and fate of
incelenmesi", to be presented in 15th National Biophysics liposomes.
Congress, Denizli-Turkey (08-12 October 2003)
Previous studies have shown that N-butyldeoxinojirimycin
(NB-DNJ), an N-glycosylation inhibitor, had a better
P272 efficiency following inclusion in liposomes as compared to
the free drug, added in the culture medium.
SERUM LIPIDS IN CHRONIC
OBSTRUCTIVE PULMONARY DISEASE The aim of this study was to investigate the intracellular
trafficking of pH-sensitive liposomes used as drug carriers
*ASLIHAN B. KARUL, **FĠSUN KARADAĞ, for NB-DNJ. We have shown that a concentration of 50
*MUSTAFA ALTINIġIK, **ORHAN ÇĠLDAĞ micromolars of liposome-included NB-DNJ decreased
*Adnan Menderes University, School of Medicine, DOPA-oxidase activity of tyrosinase to 57% as compared
Department of Biochemistry, to 95% activity in B16-F1 cells incubated with the same
** concentration of free NB-DNJ. Western-blot analyses of
Adnan Menderes University, School of Medicine, tyrosinase have shown that, in the presence of 50
Department of Chest Diseases, Aydin, Turkey micromolars liposome-loaded NB-DNJ the formation of
In earlier studies, body mass index (BMI) of chronic complex glycans is prevented and tyrosinase migrates at
obstructive pulmonary disease (COPD) patients was found lower molecular weight.
lower than healthy controls and serum total cholesterol was We have also performed in vivo fluorescent microscopy
inversely associated with hospitalization and death due to experiments using both a lipid membrane and internal,
respiratory diseases. But there is no adequate data on aqueous compartment markers, to visualize intracellular
serum lipids in COPD patients. The aim of this study is to trafficking of pH-sensitive liposomes in B16-F1 and
evaluate serum tryglyceride (TG), total cholesterol (Chol), MDBK cells. We found out that the liposomes enter into
Turk J Biochem, 2003; 28(3), 62-224 196 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
the cells via the endocytic pathway and the liposome- P275
encapsulated material is released into the cytoplasm. After
24 hours liposome lipids partially colocalized with the
ADVANCED GLYCATİON ENDPRODUCTS
Golgi apparatus in MDBK cells. (AGES) İN EXPERİMENTAL DİABETİC
NEPHROPATHY: IS SUPPLEMENTATİON
Taken together our results suggest that the pH-sensitive
liposomes cross the plasma membrane and deliver their
WİTH THİAMİNE PYROPHOSPHATE OR
content to the cytoplasm and to the secretory pathway. PYRİDOXAL 5’PHOSPHATE BENEFİCİAL?
Suat H. KÜÇÜK1,2 , M. Mert ÖZDOĞAN1, Gülden
BURÇAK1
P274 1
Departman of Biochemistry, CerrahpaĢa Medical Faculty,
COMPARISION OF MDA LEVELS Ġstanbul University, 2Centro laboratories, Ġstanbul, Turkey
MEASURED BY USING TWO DIFFERENT Increased advanced glycation end product (AGE)
HPLC DETECTORS CONCURRENTLY formation is the major mechanism implicated in diabetic
Memduh Bülbül*, Hilal Koçdor*, Zahide Çavdar*, nephropathy (DN). Limiting the rate of AGE formation has
Mehmet Eminoğlu*, Mehtap Yüksel*, Halil Resmi*, Gül been suggested as a new theraupeutic approach in DN.
Güner* Thiamine pyrophosphate (TPP) and Pyridoxal 5‘Phosphate
(PLP) have been shown to inhibit advanced glycation in
gul.guner@deu.edu.tr vitro. In this study we firstly questioned for their benefits
*Dokuz Eylül University, Learning Resources Center in DN.
Research Laboratory (ARLAB), Wistar albino male rats (n=62) ageing 8 months were
35340 Ġnciraltı-Ġzmir allocated to ―diabetic‖, ―diabetic+TPP‖, ―diabetic+PLP‖,
―diabetic+insulin‖, ―control‖, ―PLP‖ and ―TPP‖ groups.
[Objectives] Malondialdehyde (MDA), a biomarker of The administered doses were as follows: STZ 70 mg/kg,
lipid peroxidation, is commonly used in conditions ip; TPP (50 mg/kg) and PLP (50 mg/kg) in drinking water
associated with oxidative stress. Since thiobarbituric acid and insulin (4 U/day, subcutan).
(TBA) reacts with many other compounds, it has been
Glucose, HbA1c and as nephropathy indices kidney
suggested that high-performance liquid chromatography
weight/body weight ratio, urinary volume, creatinine
(HPLC) seperation might be more spesific, providing a
clearance (GFR), microalbuminuria and
relevant assay for MDA. In this study, we compared the
β2microglobulinuria were measured. AGE-peptides were
linearity of calibration curves and the reproducibility and
measured in plasma and kidney, and the activity of aldose
the recovery of a MDA method with organic phase step by reductase (AR), an enzyme for detoxification of reactive
using two different detectors, UV-VIS and fluorenscence dicarbonyl compounds was measured in the kidney.
detectors.
The data revealed the establishment of nephropathy in the
[Methods] UV-VIS detector (=532 nm) and fluorescence diabetic rats. AGE-peptides were observed to be
detector (Ex=515 nm, Em=553 nm) were connected in significantly increased both in the kidney and in the plasma
series in our HPLC procedure. Erythrocytes were used in of diabetic rats. Plasma and kidney AGE-peptide values
the study. To eliminate the effect of interfering substances, were correlated. Insülin treatment caused significant
pyridine-butanol extraction step was performed. The peaks decreases in all parameters except renal hypertrophy and
of TBA-MDA complex were obtained within 4.988 plasma AGE-peptide levels. PLP supplementation
minutes with UV-VIS detector and within 5.003 minutes improved microalbuminuria and caused nonsignificant
with fluorescence detector. The sensitivity of two detectors reductions in AGE-peptide levels of plasma and kidney.
and the linearity of calibration curves were compared. The TPP supplementation had not any effect. AR activity didn‘t
reproducibility (n=15) was calculated on days 1, 2 and 3. displayed any sinificant difference between the groups.
The recovery (n=10) was calculated at concentrations of 5, In conclusion, PLP treatment slowed the progression of
20 and 50 mol/L. diabetic nephropathy and decreased glomerular injury.
[Results] The correlation coefficients of UV-VIS and PLP supplementation may prevent the AGE-related
fluorescence detectors in the graphics were 0.99518 and damage in diabetic nephropathy. Plasma AGE-peptide
0.985388, respectively. The results obtained from levels may be considered as a marker for tissue AGE
erythrocytes were found to be within the range of 20-30 levels in diabetic rats.
mol/L and the recovery was 96% in this range. The intra-
assay reproducibility was 10%. The intra-assay variation P276
and the inter-day variation of the retention time of TBA-
MDA peaks were 1.71% and 1.14%, respectively. COMPARISON OF URINE SEDIMENTS BY
[Conclusion] The most important difference between TWO DIFFERENT METHODS
chromatograms of two detectors was that the area obtained Oytun PORTAKAL, GülĢen HASÇELĠK
with fluorescence detector was 10 times larger than the Clinical Pathology Laboratory, Hacettepe University
area obtained with UV detector for the same MDA Medical School 06100 Ankara/TURKEY
concentrations. Our results showed that both detectors
could be used successfully. However, the fluorenscence oytun@hacettepe.edu.tr
detector appeared to be more sensitive for the samples with The present study was designed to compare urine
low MDA levels. sediments of the patients by two different methods in
Turk J Biochem, 2003; 28(3), 62-224 197 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
Clinical Pathology Laboratory of Hacettepe University homocystinuria, galactosemia, biotinidase deficiency,
Medical School from November 2001 to December 2001. congenital adrenal hyperplasia and sickle cell disease and
We compared UF-100 (ROCHE, Germany) with IRIS-900 also a separate program for detection of amino acid , fatty
(DPC, USA) and manual method as a gold standart. This acid oxidation and organic acid metabolic disorders by
study was carried on five following days and one hundred tandem mass spectrometry . It uses certified DBS materials
urine samples examined per day to detect RBC, WBC, and consists of two DBS distribution components: Quality
calcium oxalate and uric acid crystals, casts and yeasts. It control (QC) materials for periodic use and quarterly
was observed 66, 34, 11 urine samples (normal, proficiency testing (PT). For the QC part, NSQAP
pathological, discordant samples, respectively) in the 1st distributes DBS materials at 6-month intervals. Participants
day, 52, 47, 9 urine samples in the 2 nd day, 49, 52, 6 urine return quantitative results from five different analytical
samples in the 3rd day, 43, 57, 8 urine samples in the 4th runs of the QC materials. The proficiency testing part of
day, 41, 54, 11 urine samples in the 5th day. Fourty seven the program provides laboratories with quarterly panels of
discordant samples were separated to examine by blind-coded DBS specimens that participants analyze once.
manually and they reexamined in IRIS-900 and in UF-100. They return their analytical results and clinical
Among 495 urine sediments, sensitivity, specificity and assessments. The program gives the laboratory an
positive predictive values were 92.6%, 91.2% and 91.1% independent external assessment of its performance .
for UF-100 and were 99.2%, 98.8% and 98.8% for IRIS- The congenital hypothyroidism and phenylketonuria
900 respectively. False negativity and false positivity were newborn screening program organized and operated by the
7.4% and 8.8% for UF-100, 0.8% and 1.2% for IRIS-900. Ministry of Health in Turkey can be achieved only through
Kappa value was 0.851. When all urine samples considered the establishment and harmonious collaboration of central
IRIS-900 and UF-100 results were observed to be and local committees and the quality assurance of the
consistent to each other. program can be guaranteed by the practice of efficient
quality control and proficiency testing programs described
P277 in the above perspectives.
QUALITY ASSURANCE PROGRAM İN [1]QC programs for thyroxine (T4), thyroid-stimulating
NEWBORN SCREENİNG hormone (TSH), phenylalanine (Phe), total galactose (Gal),
Yahya LALELĠ 17 alpha-hydroxyprogesterone (17-OHP), leucine (Leu),
methionine (Met), tyrosine (Tyr), valine (Val), and
Düzen Laboratuvarlar Grubu, Ankara/TURKEY
citrulline (Cit). We recently began offering QC materials
ylaleli@duzen.com.tr for acylcarnitines (C2, C3, C4, C5, C6, C8, C14 and C16).
Newborn screening using dried-blood-spot (DBS) collected [2]
at birth for identification of biochemical or other inherited [3]PT programs for T4, TSH, Phe, Gal, 17-OHP, Leu, Met,
conditions can effectively prevent the mental retardation, biotinidase, galactose-1-phosphate uridyltransferase, and
other disabilities and/or death associated with these sickle cell disease (SCD) and other hemoglobinopathies.[4]
disorders. Factors such as the public health infrastructure,
the financial resources available, the technological
capabilities, the differences in disease prevalences and P278
even the public awareness of newborn screening have all
affected panel of diseases covered in newborn screening
RELEVANCE OF PROCALCITONIN AS AN
programs and there is yet no uniform universal newborn EARLY INDICATOR OF SEPSIS
1
screening program upon which a consensus has been Oytun PORTAKAL, 2Nuriye FIġGIN
made. 1
Hacettepe University Medical School, Clinical Pathology
The published newborn screening program guidelines Lab. 06100, Ankara/ TURKEY
define a six part system of education, screening, follow-up, 2
Ankara Numune Education and Research Hospital,
diagnostic confirmation, treatment/management and Department of Infection Disease and Clinical
evaluation. The application of quality assurance to the Microbiology, Ankara / TURKEY
screening component will be the subject of this
presentation. oytun@hacettepe.edu.tr
A successful newborn screening program should produce The main goal of the present study was to outline the
accurate and timely reported results, should avoid missing efficacy of procalcitonin (PCT) at early diagnosis of sepsis
cases (false negative) and should minimize false positive with compared to C-reactive protein (CRP) in an adult
results that can cause parental anxiety. The means for a intensive care unit. Thirty patients who were diagnosed
laboratory to maintain and enhance the quality of its test with sepsis and SIRS according to the American Collage of
results is to participate in a quality assurance and Chest Physicians/ Society of Critical Care Medicine
proficiency testing program and to document its practice criteria were participated into the study. Patients were
in quality assurance. Our laboratory participates in the classified.as sepsis- group (sepsis, severe sepsis and septic
Newborn Screening Quality Assurance Program (NSQAP) shock, n=19), SIRS-group (sepsis origin or not, n=11) and
operated by Centers for Disease Control . The program control-group (not sepsis or SIRS, n=13). Serum
provides quality control and proficiency testing DBS concentrations of PCT and CRP were determined within 24
material for detection of congenital hypothyroidism, h after clinically onset of diseases. PCT levels were 7.8
phenylketonuria, tyrosinemia, maple syrup urine disease, ng/ml, 96.4 ng/ml, 0.7-403 ng/ml (median, mean, min-max
Turk J Biochem, 2003; 28(3), 62-224 198 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
levels, respectively) in sepsis-group, 3.8 ng/ml, 19.4 high risk group will be cost effective both for the family
ng/ml, 0.94-144.3 ng/ml in SIRS- group and 0.52 ng/ml, and for the society in the long run.
0.56 ng/ml 0.1-1,7 ng/ml in control-group. CRP levels 2. Quite a number of treatable IEM can be rapidly
were 110.0 mg/L, 94.5 mg/L, 0.0-171 mg/L (median, diagnosed from a very simple sample , namely a dried
mean, min-max levels, respectively) in sepsis-group, 72.0 blood spot which is both easy to obtain, to transport and to
mg/L, 67.4 mg/L, 0-169.0 mg/L in SIRS-group and 0.01 store. This advantage should be made use of for screening
mg/L, 0.043 mg/L, 0.0-0.5 mg/L in controls. While a IEM especially in states of emergency and in cases where
significant rise was observed for PCT (p0.05) tests are not readily available.
in SIRS-group. For CRP, a significant increase was found
in both sepsis and SIRS-groups (p0.05) both in breast 120
cancer patients and healthy subjects. However, 100
comparison of mean elemental contents of the breast
80
cancer patients with controls shows a significant
enhancement of Cu (p10 yr), and 12 male sedentary
Celal Bayar University, School of Pysical Education and controls (CG) participated in the study. Baseline serum
Sports, Manisa total PSA and Free PSA levels of the participants were
Background: Physical activity has been suggested to be measured by electrochemiluminescence immunoassay.
one of the determinants of bone turnover and to prevent Group baseline comparisons were made using a Kruskal-
age-related bone loss. To examine this we measured the Wallis test.
serum levels of osteocalcin, alkaline phosphatase, acid Results: MA and RA had higher levels of VO2max, lower
phosphatase, phosphorus, calcium and parathyroid percent body fat (p10 yr), and 12 male sedentary groups and CG in VO2max, percent body fat, and BMI
controls (CG) participated in the study. Baseline serum (except RA vs. CG) suggest that those who engage in a
calcium, phosphorus, alkaline phosphatase, and acid
Turk J Biochem, 2003; 28(3), 62-224 208 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
lifestyle of regular endurance exercise have a more P301
desirable metabolic fitness than inactive people. That there
was no statistical significance in total and free PSA levels
INHIBITION of HUMAN CATALASE by
among the three groups, gives the idea that exercise and TRIARYLMETHANE DYES
also different type of exercise alone does not have a role in Eda TOPALOĞLU, Gülberk UÇAR
reducing risk for prostate cancer. Some other factors
Hacettepe University, Faculty of Pharmacy, Department of
affecting PSA levels (having health consciousness, i.e.
Biochemistry, 06100, Ankara/TURKEY
being non-smoker, not taking alcohol and taking a healthy
diet) are as important as physical activity. Nevertheless, gulberk@hacettepe.edu.tr
more research is warranted to define further its clinical Triarylmethane (TAM+) dyes are extensively used in inks,
implications. used as a dye for wood, silk, and paper, used as a
biological stain, a blood purging agent in blood
P300 transmission, as microbicide, and anthelmintic, and also
DOES OBESITY ALTER SERUM LIPIDS IN used traditionally as an antifungal agent in aquaculture
CHILDREN? although they are not currently approved for use by the
1 Food and Drug Administration (FAD). Since TAM+ dyes
Cevval ULMAN, 1Fatma TANELĠ, 2 Betül ERSOY, have been recently shown to interact with plasma proteins
2
Dilek YILMAZ, 1Özlem TUNCER, 1Zeki ARI, 1Bekir and cellular components to cause some irreversible redox
Sami UYANIK changes in their targets possibly mediated by TAM+-
1
Celal Bayar University, Faculty of Medicine, Dept of derived free radicals and to have toxic and carcinogenic
Biochemistry, Manisa, Turkey effects on mammalian tissues, three TAM+ dyes, malachite
2
Celal Bayar University, Faculty of Medicine, Dept of green (MG+), leucomalachite green (LMG+) and gentian
Pediatrics, Manisa, Turkey violet (GV+), were tested for their inhibitory actions on
human catalase. Catalase, one of the key protective
ulman@isbank.net.tr enzymes against the reactive oxygen species (ROS)
Obesity is generally accepted as a risk factor for produced in the cell, was isolated from human erythrocytes
atherogenesis for adults, and it is frequently accompanied with a spesific activity of 160 U/mg. The Km and the Vmax
by increased serum lipids. To test whether this is also true values of the crude enzyme were found as 27 mM and 200
in children, we conducted a case control study in a group mmol/min/mg protein, respectively. All of the TAM+ dyes
of children in our region. tested inhibited the human catalase non-competetively and
irreversibly by incubating the enzyme with inhibitors at the
We assessed the relation between level of obesity and
various concentrations for 0-60 minutes at 370C. Ki values
apolipoproteins (Apo A-I, ApoB), Lipoprotein (a) (Lip(a),
were found as 8-20 mM at these conditions. The enzyme
and serum lipids. The study group 7.5-17.2 years old
was completely inactivated with the relatively high
(mean 12.9yrs), included 21 obese children, and 83
concentrations of MG+ and LMG+ up to 40 mM . Although
controls (male/female: 53/52). The groups were similar
the mode of the inhibition of catalase with TAM+ dyes
regarding age and sex, but body mass index (BMI) was
appeared as non-competitive, the mechanism seemed
significantly higher in the obese children.
complex. These preliminary results suggested that
In the non-obese children, Apo A-I levels positively irreversible inhibiton of erythrocyte redox enzymes by
correlated with total cholesterol, high (HDL) and low- TAM+ dyes which might lead an increase in the ROS
density lipoprotein (LDL) cholesterol, but Apo B levels production and lipid peroxidation in the cell could play an
correlated only with cholesterol and LDL cholesterol. In important role in cell demage in human.
the obese children, Apo A-I levels correlated only with
cholesterol, and ApoB levels positively correlated with
cholesterol and LDL cholesterol. Lip (a) levels did not P302
correlate with cholesterol, trigliserid, HDL cholesterol and EFFECT OF STATIN TREATMENT ON
LDL cholesterol in obese and non-obese children
INSULIN-LIKE GROWTH FACTOR IN
The Apo A-I, and ApoB levels were not different between POSTMENOPAUSAL WOMEN
obese and non-obese boys, while these were significantly 1
lower in obese girls (p=0.000, p=0.003 respectively). Fatma TANELI, 2Canan TIKIZ, 1Cevval ULMAN, 2Zeliha
However the Lip(a) and Apo A-I:B ratios were not ÜNLÜ, 3Hakan TIKIZ, 1Bekir Sami UYANIK, 2Çiğdem
different between study groups in either sex (p >0.05 ). TÜZÜN
Apo A-I, Apo B, and Lip (a) levels did not correlate with Celal Bayar University, Faculty of Medicine, Departments
the level of obesity (BMI) in obese and non-obese children. of 1Biochemistry, 2Physical Medicine and Rehabilitation,
3
Cardiology, 45020, Manisa/Turkey.
These results suggest that, most of the serum parameters
used classically in the assessment of the risk for fatma.taneli@bayar.edu.tr
atherogenesis is not pertinent in childhood. Obesity may Insulin-like growth factor-I (IGF-I) is an essential factor
not necessarily alter these parameters in childhood, and for longitudinal bone growth and stimulation of both
making prospects for the prevention of early onset proliferation and differentiation of osteoblasts. Early
atherogenesis in adulthood becomes difficult. Thus, epidemiologic studies examining the association of 3-
research for new parameters to assess the risk in children is hydroxy-3methylglutaryl coenzyme A reductase inhibitors
required. (statins) in preventive therapy of osteoporotic hip fractures
Turk J Biochem, 2003; 28(3), 62-224 209 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
produced encouraging results. In the present study, we Our results suggest that saliva leptin concentrations may be
aimed to investigate the early serum changes in IGF-I and used clinically instead of serum leptin concentrations
IGF binding protein-3(IGFBP-3), which is the major because of being sample collection not invasive
binding protein of IGF-1, levels before and after three
months of statin medication. Thirty women with untreated
postmenopausal osteoporosis were taken into the study. P304
Blood samples were obtained before and after 3 months of EFFECTS OF SUBCHRONİC TREATMENT
statin treatment. Serum IGF-I and IGFBP-3 levels were
assessed by enzyme-linked immunosorbent assay method
OF THİOCARBAMİDE ON
by DSL (Diagnostic Systems Laboratories, Inc. Webster, HAEMATOLOGİCAL AND BİOCHEMİCAL
Texas, USA) reagents. Bone turnover markers of CONSTİTUENTS OF RATS
osteocalcin, parathyroid hormone, and C-telopeptide of Ġsmail ÇELĠK, PhD Hanefi ÖZBEK, MD, PhD * and
type 1 collagen (CTX) levels were assessed on serum Yasin TÜLÜCE
samples by automated chemiluminescence method by
Yuzuncu Yil University, Science and Art Faculty,
commercial reagents on autoanalyzer (E170 Modular
Deparment of Biology, Van, Turkey
System, Roche Diagnostics Corporation, Indianapolis,
USA). Total cholesterol, triglyceride, HDL cholesterol, *Yuzuncu Yil University, Medicine Faculty, Deparment of
LDL cholesterol, calcium, phosphorus and total alkaline Pharmacology, Van, Turkey
phosphatase were assessed by enzymatic methods on The effects of sublethal concentration of Thiocarbamide on
autoanalyzer (Integra Roche Diagnostics Corporation, various haematological and biochemical costituents of rat
Indianapolis, USA). Bone alkaline phosphatase was was investigated under laboratory conditions. 250-ppm of
assessed by heat inactivation method. Bone mineral density Thiocarbamide was administered orally to 8 rats ad libitum
was assessed by dual energy X-ray absorbtiometry. We during the tests for 25 days cosecutively.
found significant (p 5.5 uIU/ml (n=9) .
Serum thyroid function tests were determined with
PHOSPHATE DEHYDROGENASE FROM electrochemiluminesans assay on ACS Centaur
SIX-MONTH-OLD LAMB KİDNEY CORTEX autoanalyser and IGF-1 levels were measured by non
Ulusu N.N., Tandoğan B. extraction IRMA.
SPSS (Version 6.0) for Windows was used for istatistical
Hacettepe University, Faculty of Medicine, Department of
analysis with Kruskall-Wallis, One way ANOVA and
Biochemistry, 06100 Ankara/Turkey
Mann Whitney U tests.
Glucose-6-phosphate dehydrogenase (D-Glucose-6-
A strong inverse relationship between age and IGF-1 levels
phosphate: NADP+ oxidoreductase EC 1.1.1.49) catalyses (r= 0.6767, p=0.000) and T3 and IGF-1 (r=0.4965, p=
the first and rate limiting step in the pentose phosphate 0.031) by Pearson correlation. We found the negative
pathway. We purified glucose-6-phosphate dehydrogenase correlation between TSH and IGF-1(r=-0.343, p=0.05) in
from six-month-old lamb kidney cortex for the first time. thirty patients. There weren‘t any correlations between
By the other authors, a variety of methods consisting of IGF-1 and gender, T4, FT3, FT4. IGF-1 levels were
numerous steps have been applied to obtain a reasonable significantly higher in Group I (668.25+/-278 ng/ml,
amount of pure enzyme from other organisms and tissues. p50% stenosis affecting at least one controls. Cell lysates were applied to % 10 SDS- PAGE
artery were included in group with coronary heart disease electrophoresis run at 50 mA for 3 hours. Proteins were
(patients) 2-15 women with < 20 % stenosis were enrolled
transfered to nitrocellulose membrane by Western blot and
in group without disease (controls). Mean ages of patients
treated with anti IPO-3 monoclonal antibody. Native gels
and controls were 64,06±8,65 and 66,12±6,80,
were also used for separation of lymphocyte proteins and
respectively. After DNA was extracted from whole blood
also detected by immunoblotting. Bands were detected by
samples with salting-out method, genotypes were
peroxidase staining.
analyzed by polymerase chain reaction-restriction fragment
length polymorphism. Statistical analyses were computed Results and discussion
by SPSS 11,5 version, using nonparametric tests. Although
In SDS-PAGE, SLAM band is found to be approximately
the prevalences of 1298CC and 1298CC/AC were higher
70 kD and in activated lymphocytes was larger as
in patients with respect to controls (p=0.009; p=0,016,
compared to that of unstimulated lymphocytes or HL-60
respectively),the significant difference was not observed
cells. This is in accordance with the stimulous dependent
in the prevalences of the other genotypes between the
expression of the molecule. SLAM yielded single bands
groups. There was the positive correlation between
under denaturing and nondenaturing conditions . This
coronary disease and the frequency of 1298CC ( r=0,447
indicates that it consists of a single chain and has no
p=0,017) The odds ratio was 1,71 (p=0,038, 95% CI, 1,00
to 2,92) in the patients with 1298CC mutation with respect subunits. Furthermore, mercaptoethanol treatment did not
to without. It was also 1,71 ( p=0,026, 95% CI, 1,00 to cause any shift in the size of the molecule. However,
2,66) for 1298 CC as compared with 1298 AA/ AC SLAM moved very slowly in native gels as compared to
combination. The high prevalence of the 1298 CC that of SDS-PAGE indicating that the glycosilated part
genotype might be effective on the genesis of the disease consists basicly of neutral glycolipids and/or the protein
itself and an important risk factor in the occurrence of core is not rich in negatively charged amino acids. It is also
coronary disease in postmenopausal women. found that immunodetection of SLAM is much more
Turk J Biochem, 2003; 28(3), 62-224 216 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
efficient when native gels are used. It is concluded that programmed cell death. One of these components,
SLAM as an immunodominant molecule may be suppresed Elongation Factor 2 (EF-2) is a protein involved in
by MV enfections and further studies on the eukaryotic polypeptide chain elongation and promotes
characterization of SLAM may reveal insights in molecular transloction in this process. In this study, actin and EF-2
immunological mechanisms. interaction was investigated and an interaction was shown
in nearest in vivo conditions using cell homogenates by
References
anti-actin antibody and protein electrophoresis,
1. Tatsuo, H., Ono, N., Tanaka, K. and Yanagi,Y., SLAM contributing to the in vitro findings of an interaction
is a cellular receptor for measles virus, Nature, 406: 893- between EF-2 and actin.
897, 2000.
2. Movaddat, N., Mason, D.W., Atkinson, P.D. et.al,
P321
Signalling lymphocytic activation molecule ( CDw150) is
homophilic but self-associates with very low affinity. J THE EXAMINATION OF THE
Biol. Chem., 275(36): 28100-28109, 2000 FUNCTIONAL PROPERTIES OF
P319
ADENOVIRUS TYPE 5 E4 ORF3 PROTEIN
Emre Koyuncu, Orkide Özge Dirlik, Thomas Dobner
MONOCLONAL ANTIBODIES AGAINST
HUMAN MYOGLOBULIN; FUNCTION Institut für Medizinische Mikrobiologie und Hygiene,
LandshuterStr. 22, 93047, Regensburg/Germany
ANALYSIS WITH BIOSENSOR
TECHNIQUES emrekoyuncu@lycos.com
Ozeren, Muserref Adenovirus is one of the mostly used gene therapy vectors
to date. However, a major problem is the oncogenic
Marmara University, Arts and Sciences Faculty, potential of these viruses. E4 ORF3 protein is one of the
Deparment of Biology, Goztepe Kampusu, Istanbul, Turkey oncoproteins of Adenovirus Type 5. Besides its oncogenic
The interaction between human heart myoglobulin and properties, previous reports have shown that it takes part in
eight specific monoclonal antibodies (mAbs) was viral DNA replication, late viral protein synthesis, and
investigated with real time biomolecular interaction shut-off of host protein synthesis. For this reason, in order
analysis (RT BIA), using Surface Plasmon Resonance to develop a more efficient and safe gene therapy vector,
(SPR). The purpose of this study was selection of high the functions of this protein should be well understood.
affinity mABs for the Nycocard, a rapid quantitative
Most of our knowledge about the functions of ORF3
immunoassay format. Analysis of association and protein comes from its mutational analysis. It has never
dissociation kinetics was monitored in real time, with been expressed and purified successfully. In this extand,
unlabelled reactants. Antibody isotyping was rapid and we tried to express ORF3 as a Glutathione-S-Transferase
simple. fussion protein in E. coli. At the end, we successfuly
Monoclonal antibodies with four different epitope expressed and purified not only the wild type ORF3 but
specificities and optimal binding function were selected for also four mutant forms, two of which lacking different
a myoglobulin sandwich assay with enhanced sensitivity. regions in N-terminus, and the other two in C-terminus.
Moreover, by GST-capture assays, we determined
interacting regions of ORF3 with the proteins that have
P320
been shown to bind ORF3 in vivo. Our results showed that,
INTERACTION OF ELONGATION FACTOR C-terminus of ORF3 was important for protein
2 WITH ACTIN interactions, at least for the proteins we have tested so far.
Kıvanç ERGEN1, Aynur KARADENĠZLĠ2, Fethiye This was also in agreement with the data we had from
KOLAYLI2, Bent RIIS3 immunoflourescent analysis. One of the mutant ORF3
1 having a single amino acid exchange in C-terminus showed
Kocaeli University Medical Faculty, Department of completely different localization from the wild type in the
Biophysics, 41900, Derince-Kocaeli, TURKEY. cell. The effects of this mutation and other mutations in the
akergen@superonline.com same region on viral growth and ORF3‘s interacting
2
Kocaeli University Medical Faculty, Department of partners is currently under our investigation. The results of
Microbiology, 41900, Derince-Kocaeli, TURKEY this research will contribute to understanding of the
3 functional properties of ORF3 which is very important for
Department of Animal Product Quality, danish Institute of
developing adenovirus vectors.
Agricultural Sciences, Research Centre Foulum, P.O. Box
50, DK-8830 Tjele, DENMARK
It has been indicated that several components of protein
synthetic machinery can bind to actin microfilaments and
their interactions with cyotoskeleton can play a role in
Turk J Biochem, 2003; 28(3), 62-224 217 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
P322
PURIFICATION AND
CHARACTERIZATION OF RAT SMALL
INTESTINE GLUCOSE-6-PHOSPHATE
DEHYDROGENASE
Ali DANIġAN, Deniz CEYHAN, Hamdi ÖĞÜġ and
Nazmi ÖZER
Hacettepe University, Faculty of Medicine, Department of
Biochemistry, 06100, Ankara/TURKEY
naozer@hacettepe.edu.tr
Glucose-6-phosphate dehydrogenase (G-6-PD) catalyzes
the first step of the pentose phosphate patway which
generates NADPH as reductive power for antioxidant
systems and anabolic patways. G-6-PD was purified from
the rat small intestine by successively use of
homogenization, ultracentrifugation (105000 xg), ion
exchange chromatography (DEAE-Fast Flow), dialysis and
affinity chromatography (2‘,5‘-ADP-Sepharose 4B) with a
specific activity of of 126 UI/mg protein and a yield of 21
%. PAGE showed three bands on protein staining, only the
fast moving band had G-6-PD activity. Km values of
enzyme for its substrates , NADP and glucose-6-phosphate,
were calculated to to be 22,5 and 61,9 micromolar,
respectively.
Rat small intestine G-6-PD has a ph optimum of 8.3. The
activation energy, activation enthalpy and Q10 for the
enzymatic reaction were calculated to be 8516 cal/mole,
7902 cal/mole and 1.59, respectively.
Turk J Biochem, 2003; 28(3), 62-224 218 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
AUTHOR INDEX
ABDULOVA G. 75 AMANVERMEZ R. 199 BARAN A. 127, 128
ABOU AUDA M.M. 98 ANACHKOVA B. 74 BARICEVIC I. 96
ACAN N.L. 207 ANANIEV E.D. 75, 144 BARIS I. 170, 215
ACAR C. 75 ANANIEVA E.A. 145 BARISKANER H. 111
ACARTURK E. 140 ANDREASYAN N.A. 94 BARKUT Y. 136
ACIK L. 113, 114 ANITA M. 188 BARLAK Y. 119
ADALI O. 178 ANLAR B. 216 BARSEGYAN K. 122
ADIGUZEL Z. 205 APOSTOLOVA D. 152 BARZU O. 74, 159
AFRASYAP L. 170, 215 ARDICOGLU A. 204, 205 BASKOL G. 199, 202
AGACHAN B. 93 ARI Z. 208 BATCIOGLU K. 103, 127
AGADJANYAN A.KH. 93 ARICAN A. 179 BATUN S. 79, 177
AGAR A. 212, 213 ARIKAN M.K. 160 BAY KARABULUT A. 128, 159, 194
AgGADJANYAN A.A. 93 ARINC E. 77, 178, 179 BAYIR Y. 165, 175
AGIRBASLI M. 183 ARNAUTOVIC A. 132 BAYRAKTAR N. 194
AGIRDEMIR H. 143 ARSLAN B. 213 BAYRAMOGLU F. 112
AGLAMIS S. 174 ARSLAN S. 178 BAYSAL K. 205
AK BASOL G. 186 ARSOVA V. 121 BAYSAL S. 127, 128
AKALIN N. 82 ARSOVSKI A. 185 BEGIC L. 132
AKAR S. 100 ARTAMONOV M.V. 94, 95 BELGE KURUTAS E. 106, 156, 157,
AKDAG M.Z. 113, 118, 141, 146 ARTENI A.A. 180 158
AKDENIZ K.G. 195 ARTENIE V. 180 BENLI AKSUNGAR F. 96
AKIN A. 86 ASLAN A. 165, 175 BERZAL-HERRANZ A. 81
AKKAN I. 82 ASLAN B. 164 BIBEROGLU G. 89
AKKAS S.B. 73 ASLAN D. 82, 201, 206 BICAK E. 127, 128
AKKAYA O. 210 ASSA P. 205 BIKMAZ B. 126
AKMEMIS C. 170 ATAMER Y. 79, 177 BIKMAZ K. 126
AKOZ M. 212 ATANASOV B. 163 BIKMAZ P.S. 126
AKPINAR P. 216 ATANASOVA L. 141 BILGI C. 73, 194
AKSEN F. 106, 113 ATANASSOV B. 74 BILGIHAN A. 122, 123
AKSOY K. 104, 174 ATAY A. 168, 211 BILGIN M.D. 81
AKSOY Y. 102 ATMACA G. 127 BINGOL B. 210, 214
AKSU A. 91 ATUKEREN P. 158, 214 BODEANU M. 76
AKSU U. 158 AVRAM S. 108 BODUR E. 191
AKTAS Z. 115, 116 AYAR KAYALI H. 171 BOGDANOVIC G. 177
AKYILMAZ E. 84, 133, 139 AYDIN B. 169 BOGOJEVIC D. 119
AKYOL O. 133, 174, 182 AYDIN E. 192 BOJIC D. 146
ALABAKOVASKA S. 131 AYDIN S. 209, 213 BOJINOVA V. 144
ALAVANTIC D. 153 AYDOGAN S. 191 BOLAMAN Z. 193
ALBENIZ I. 126 AYDOGDU N. 127, 147, 148 BOR-KUCUKATAY M. 212
ALCHUJYAN N. 122 AYGIT A.C. 114, 115 BOSTANCI M. 82
ALEXANDROV I. 158 AYGUN S. 190 BOYANOV A. 149
ALEXANDROVA R. 158 AZIBI K. 76 BOYANOVA M. 81
ALEXANDRU D. 156 • BOYUNEGMEZ T. 179
ALICIGUZEL Y. 213 BABEANU C. 120 BOZCARMUTLU A. 179
ALP A. 207 BAGIYEVA S. 105, 141, 215 BOZKURT A. 140
ALTANER C. 158 BAIDANOVA V. 144 BREWER J.M. 160
ALTANEROVA V. 158 BAKALOVA N. 131, 135 BROCK J.H. 155, 160
ALTANKOV G.P. 166 BAKALOVA R. 105 BROWN K.A. 176
ALTINISIK M. 195 BAKAN E. 179 BUBER E. 207
ALTUNDAL H. 169 BALAN S.A. 155 BUGARINOVIC S. 176, 183, 184
ALVER A. 160 BANKOVA E. 131 BULBUL M. 196
Turk J Biochem, 2003; 28(3), 62-224 219 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
BURCAK G. 197 COSKUN G. 137 DONMA O. 100
BURTEA M. 155 COSTACHE G. 76 DORAN F. 150
BUYUKBESE M.A. 106 CREANGA D.E. 180, 188, 194 DORDEVIC A. 177
BUYUKGEBIZ B. 91, 203 CREANGA I. 180 DRAGIC M. 121
BUYUKYAZI G. 207, 208 CUTTONE G. 85 DRAGOMIR E. 83
• • DRAMICANIN T. 132
CABA A. 102 DAGLAR A. 113, 147, 148 DROSOPOULOS C. 80
CABADAK H. 169 DALDAL F 190, 210 DUMAN E. 134, 154, 153
CAKIR A. 165, 175 DALKARA T. 78 DUNDAR E. 140
CAKIR B. 114, 115 DALKILIC N. 111, 112 DURUKAN H. 78
CAKIR D.U. 79, 141, 146, 147, 177 DALMIZRAK O. 78 DUZGUNCINAR O. 193
CAKIR E. 73, 197 DANISAN A. 217 •
CALISKAN S. 214 DAPCEVIC B. 146 ECONOMOU A. 99
CALISKANER P. 168 DAS D. 99 EFREMOV G.D. 71, 75
CAN YILMAZTURK G. 82 DASDAG S. 113, 118 EKERBICER N. 195
CANACANKATAN N. 150 DEDOVIC N. 132 EKMEKCI O.B. 100
CANDAN N. 161 DEGER O. 119, 160 ELBAKYAN G. 122
CANEVARI S. 129 DEJANOV P. 186 ELDEM Y.Y. 130
CANIC V. 176 DEJANOVA B. 185, 186 ELIBOL B. 78
CANORUC N. 79, 177 DEMAJO M. 85, 105, 106, 121, 168 ELMAS O. 213
CANSEVEN A.G. 122, 123 DEMIR T. 100 EMINOGLU M. 196
CAPRARU D. 83 DEMIRCEYLAN E. 175 ENGIZEK T. 172
CAPRARU M. 83 DEMIRCI C. 214 ENLI Y. 201, 206
CAVDAR Z. 196 DEMIRCIOGLU A.S. 183, 184 ERARSLAN A. 143, 162
CELEBI A. 113, 114 DEMIREL H. 100 ERBAS H. 113, 148, 147
CELIK C. 199 DEMIREL I. 111, 112 ERCAN A. 78
CELIK I. 210 DEMIREVSKA-KEPOVA K. 110, ERCIYAS F. 211
CELIK O. 187 111 EREM C. 160
CELIK T. 73, 166, 194 DEMIRKAZIK A. 150, 165 EREN A. 96
CERINIC M. 99 DEMIROGLU A. 202 EREN N. 164
CETIN A. 189 DEMIRPENCE E. 193 ERGEN H.A. 93
CETINKAYA A. 106 DENES A.S. 74, 159 ERGEN K. 216
CETINKAYA H. 214 DENIZCI A.A. 143, 162 ERGENC S. 172
CEYHAN D. 217 DERIN N. 213 ERGIN H. 201, 206
CHELU F. 155 DEVECI R. 186 ERMAN F. 209
CHERNEV P. 86, 172 DIMITRIJEVIC B. 121, 132, 151 ERSOY B. 208
CHISTOV K.N. 145, 171 DIMITROV I. 145 ESKANDARI H. 92
CHRISTOU A. 187 DIMITROVA R. 81 EVRAN S. 137
CHRISTOV C. 143 DIMITROVSKI C. 75 EYSKENS F. 90, 203, 204
CHUBARYAN S.V. 93 DIMOVSKI A.J. 71 •
CICIK E. 100 DINCER A. 72, 137, 138 FADILLIOGLU E. 174
CIGERLI S. 164 DINCKAYA E. 84, 133, 136, 137, 139 FARBER D.B. 78
CIGREMIS Y. 187 DINIC S. 119 FEDINA I. 98, 103, 109
CIKIM G. 157 DINU D. 118, 131 FELLER U. 110
CILDAG O. 195 DIRLIK O.O. 217 FERRI R. 129
CIOBANU G 120 DJUJIC I. 168 FIGINI M. 129
COKUGRAS A.N. 191, 192 DOBNER T. 217 FILIP C. 102, 103
COLAK M. 119 DOBRIKOVA A.G. 136 FILIPCE D. 186
COLIC N. 146 DOBRIN A. 76, 155 FILIPCE V. 185, 186
COLOMBO M. 129 DOGAN A. 206 FILIPCE V.I. 185
COMAN A.C. 97, 129 DOGAN A.L. 193 FILIPOVIC D. 101, 139
CONDAC E. 118 DOGAN Ayca 193 FISGIN N. 198
CONSTANTINESCU E. 126 DOGAN Ayse 150, 165 FRANGOPOL P. 187
CORIC J. 190 DOGAN N. 111 FUSTIK S. 129
CORUH N. 189, 192, 200 DONEVSKA J. 186 •
Turk J Biochem, 2003; 28(3), 62-224 220 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
GADJEVA M. 114, 163 HACIOGLU G. 212 JURANIC Z. 167
GANEA E. 96, 97, 118, 129 HADJIMITOVA V. 105 •
GANEV V. 114 HAIRAPETYAN H.L. 94 KABALAK T. 153, 154
GARGILI A. 214 HALICI M. 165, 167, 175 KACAR S.B. 86
GATEVA P. 94 HALIFEOGLU I. 204, 205, 209 KAHRAMAN H. 205
GEGOVA A. 109 HARDALAC F. 124 KALFIN R. 99
GENC K. 210 HARDING J.J. 96 KALKAN M.T. 162
GENC M. 103 HASCALIK M. 187 KAN B. 169
GENC S. 210 HASCALIK S. 187 KANDIL A. 214
GEORGIEV M. 94 HASCELIK G. 197 KANIT L. 138
GEORGIEVA K. 98, 103, 109 HATZISTAVROU E. 98 KANJER K. 76
GEZER S. 156 HAZAROSOVA R. 151 KAPLAN A. 79, 177
GHAZARYAN S. 122 HAZNECI E. 103 KAPTANAGASI ORCUN A. 142
GHEORGHITA N. 102, 103 HEKIM N. 89, 184 KARAALI Z.E. 93
GLISIC S. 153 HEKIMSOY Z. 168 KARAARSLAN I. 88, 198
GLODEANU E. 120 HELTIANU C. 75, 155 KARACALI S. 186
GOCER F. 165 HERLYN D. 129 KARADAG F. 195
GOKER B. 169 HLUBINOVA K. 158 KARADENIZLI A. 216
GOKER I. 154 HOLZER R. 110 KARAGYOZOV L. 75
GOKKUSU C. 126 HORVAT A.I. 105, 121 KARAHAN S.C. 160
GOKTUG T. 136, 137 HORVATH A. 114 KARANFILSKA D.P. 71, 75
GOLDFINE H. 190 HOSVER I. 160 KARAVELIOGULLARI F.A. 114
GOLTSEV V. 86, 172, 173 HRISTOVA M. 176 KARDESOGLU E. 166
GORGULU G. 72, 139 • KARSHIKOFF A. 71, 81
GORGULU S. 72 ILHAN B. 111 KARTAL F. 135
GRIGOROVA I. 98, 103 ILHAN Necip 204, 205 KARUL A.B. 195
GROZDANOV P. 75 ILHAN Nevin 204, 205 KASAPOVIC J. 101, 139
GUDUMAC V. 128 ILIC M. 146 KAYIR H. 73, 194
GUIDUCCI S. 99 ILIC Z.S. 144 KAYMAK K. 127, 148, 147
GUJA C. 155 ILKAY E. 205 KAYNAR O. 194
GUL S. 156, 158 INANC F. 106, 156, 157, 158 KAYRIN L. 150
GULA N.M. 94, 95 INHAN G.A. 162 KAZAN D. 143, 162
GULDUR T. 194 IOCHEV M. 85 KAZANCI N. 170, 171, 213
GULEC M. 133, 174, 182 IONESCU-TIRGOVISTE C. 155 KAZAZ C. 175
GULEN S. 113, 114, 115 IPCIOGLU O.M. 166, 199 KAZEZOGLU C. 115
GULER G. 124 IPEK E. 140 KELES S. 167
GULTEPE M. 166, 199 IRAZ M. 133, 174, 182 KELESTIMUR F. 91
GUMUSTAS M.K. 158, 214 IRFANOGLU M.E. 113 KENGIL I. 96
GUNAY I. 150, 165, 182 ISBIR T. 93 KERIMAK ONER M.N. 162
GUNDOGDU U. 142 ISCAN M. 200 KESKIN S. 207
GUNEBAKAN S. 172 ISGOR B. 200 KEVORKIAN G. 122
GUNEL-OZCAN A. 178 ISIK B. 113 KHMEL T.O. 95
GUNEN H. 159 ISLEKEL H. 214 KILIC E. 202
GUNER G. 155, 175, 196 IVANOV I. 81, 123, 158 KILIC N. 209
GUNGOR O. 116 IVANOV R. 189 KILINC A. 135
GURAY T. 72, 139 IVANOVIC-MATIC S. 119 KILINC E. 156
GURBILEK M. 212 • KILINC M. 106, 156, 157, 158
GURKAN H. 107 JAKIMOV D. 177 KILINC N. 118
GUVEN A. 157 JANAC B. 180 KIRAN H. 157
GUVENC H.B. 205 JANEZIC D. 85 KISHORE U. 114, 163
GUZEL S.P. 209 JOKANOVIC M. 151, 180 KIZIL C. 139
GUZELDEREN F.G. 100 JORDANOVA A.G. 112, 119, 160 KJURKCHIEV S. 167
GUZVIC M. 132 JORDANOVA P. 158 KNEZEVIC-USAJ S. 121
• JOVANOVIC Z. 153 KOCABAS V. 212
HACIEVLIYAGIL S.S. 159 JOZANOV-STANKOV O. 151, 168 KOCDOR H. 196
Turk J Biochem, 2003; 28(3), 62-224 221 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
KOCER M. 159 MANDUTEANU I. 83 NICHITA N. 149
KOCEVA B. 75 MANEA A. 126 NICOLAE M. 156
KODROV P. 85 MANNERVIK B. 102 NIKETIC V. 116, 117
KOJIC V. 177 MANUKYAN A.V. 93 NIKOLAVA A. 143, 144
KOJOUHAROVA M. 114, 163 MARDANYAN S.S. 94 NIKOLIC J.A. 96
KOLAROSKA D. 101 MARINESCU G. 120 NIKOLIC M. 116, 117
KOLAYLI F. 216 MARKICEVIC M. 152 NIKOLIC-VUKOSAVLJEVIC D. 76,
KOLDAS L. 100 MARKOV D.V. 98 152
KOLEV D. 131, 135 MARKOVA M. 95 NIKOLOVSKA E. 186
KOLEVA I. 109 MARKOVIC L. 168 NIKOLOVSKI M. 94
KONUS M. 200 MASALCI O. 171 NIKOYAN A.A. 93
KORICANAC G. 78 MASLENKOVA L. 109 NINIO S. 112, 119
KORICANAC L. 85, 106 MEDIC L. 151 NIWA T. 81
KORKMAZ F. 211 MELANI C. 129 NOVAKOVIC J. 152
KORKMAZ M. 205 MEMISOGULLARI R. 167 •
KORUGAN U. 91 MENZILETOGLU S.Y. 104 OBA R. 183
KOSEOGLU M.H. 168 METE N. 141, 146 OCAL I. 150, 165
KOSTOVA N.N. 98 MIHAILOVIC M. 119 OD Z. 95
KOUZMANOVA M. 176 MIKOSHA O.S. 95 ODABASOGLU F. 165, 175, 167
KOYUNCU E. 217 MILAC A. 108 ODACI D. 133, 134
KOZELOV L. 148 MILEVA K.N. 189 ODJAKOVA M. 167, 168, 173
KRSTIC-DEMONACOS M. 109 MILLER L.M. 102 OGUS A. 75, 141, 215
KRTOLICA K. 121, 132 MILOVANOVIC S. 124 OGUS H. 78, 217
KUCUK S.H. 197 MINCHEVA J. 143, 144 OK E. 202
KUCUKATAY V. 212 MIRONOVA R. 81 OKABE H. 155, 175
KULAKSIZ G. 78 MITREVSKA S. 101 OKAJIMA K. 155, 175
KUNDAK A.A. 148 MIZRAK B. 187 OKAY S. 140
KUNEVA T. 152 MLADENOV E. 74 OKUR E. 157
KURAL A. 164 MLADNOV D. 94 OKUTUCU B. 72, 117, 138
KURALAY F. 210, 214 MODEVA T. 148 OLGUN A. 188
KURKCUOGLU M. 140 MOMIC T. 140 ONAL S. 127, 128, 153, 154
KURTUL N. 179 MOUMTZI S. 80 ONCUL S. 162
KUTAY F.Z. 186 MOUSTAKAS M. 187, 188 ONER C. 75, 78, 141, 215
KYRIAKIDIS D.A. 87 MOVSESYAN H. 122 OPRESKO L. 108
• MOVSESYAN N. 122 ORHAN B. 207
LABUDOVIK D. 131 MRSA V. 82 OS M. 95
LALCHEV Z.I. 112, 119, 160, 166 MUFTUOGLU M. 78 OSMAN O. 154
LALELI Y. 197 MUHTAROGLU S. 199, 202 OZALP M. 207
LAMBEV I. 105 MUJAGIC H. 120 OZAND P.T. 71, 87
LAMBREV P. 86, 173 MUJAGIC Z. 120 OZAR M.O.K. 172
LARSSON A.K. 102 MURGULET C. 194 OZBEK H. 210
LASTUN V. 150 MUSTATA R.C. 196 OZBEK M. 195
LAVCANSKA J. 121 MUTLU L.C. 159 OZCAN E. 96
LEAHU R. 188 • OZCAN O. 166, 199
LEFEVERE M. 203 NACHEVA G. 81 OZCAN ORUC E. 164
LUKAREVSKA V. 129 NADOLNIK L.I. 122, 125 OZCELIK F. 115, 116
LUPACHYK S.V. 122 NAIRA S. 108 OZDOGAN M.M. 197
LYGEROU Z. 77 NANEV C. 145 OZEL U. 122, 123
• NECHIFOR M.T. 118, 131 OZEN H. 207
MAFTEI M. 102 NECHIFOR Mihai 103 OZER I. 130
MAHURAN D.J. 181 NEDELJKOVIC J. 96 OZER N. 78, 191, 192, 217
MAJKIC-SINGH N. 146, 153, 162 NEDEVA D. 143, 144 OZEREN M. 216
MALESKA V. 186 NEDIC O. 96 OZEROL E. 174
MANCEVSKA R. 186 NEICHEVA T. 112, 119 OZGEL N. 193
MANDACI S. 190, 210 NIATSETSKAYA Z.V. 122, 125 OZGUL K. 75
Turk J Biochem, 2003; 28(3), 62-224 222 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
OZGUL R. 78, 105, 141, 161, 215 PISSARSKA M. 141 SAHIN I. 170, 213
OZHAN H.G. 139 PLASESKI T. 75 SANTER R. 129
OZKAL S. 214 POENARU L. 76 SARAYMEN R. 202
OZKAN S. 100 POPDIMITROVA N. 109 SARDOGAN E. 100
OZKAN Y. 209 POPHRISTOVA E. 109 SARGISOVA Y.G. 94
OZKARA H.A. 92, 181, 137 POPOV D. 126 SARGSYAN N. 108
OZON A. 88 POPOVA L. 145, 171, 181 SARIBAS Z. 207
OZSAVCI D. 183, 184 POPOV-CELEKETIC D. 76, 151 SARICAOGLU S.M. 105, 141, 215
OZTURK I.C. 103 POPOVSKA Z. 186 SAVCIOGLU F. 212
OZTURK N. 87, 185 PORTAKAL O. 197, 198 SAYERS Z. 200
OZTURK T. 135 POTURJANOVA M. 158 SAYLI O. 86
OZTURK UREK R. 135 POZNANOVIC G. 119 SCETA S. 190
OZTURK Y. 210 PROFIROV Y. 148 SECKIN C. 200
OZUTEMIZ O. 214 PRTENJAK G. 184 SECKIN D. 204, 205
OZYUREK N. 164 PRUNESCU C.C. 155 SEKBAN H. 142
OZYURT H. 133, 182 PRUNESCU P. 155 SEMENOV E. 85
• PSAHOULIA F. 80 SEMIZ S. 201
PAJOVIC S.B. 101, 139 PSARRAS I. 80 SENER A. 169
PAK W.L. 85 PUNEVA I. 143 SERBAN-CAPATINA A.I. 118
PALA F.S. 107 • SERT C. 141, 146
PALA O. 107 RADETA M. 152 SERTER M. 193
PANCHEV I. 167, 168 RADEVIC B. 146 SEVEN S. 207
PANKOV R.G. 166 RADICHEVA N.I. 189 SEVERCAN F. 72, 73, 139, 170, 189,
PANTAZAKI A.A. 87 RADIVOJSA S. 78, 124 206, 211, 213
PAP D. 176, 183, 184 RADOJCIC M.B. 101, 139 SEYHAN N. 122, 123
PAPADOGIANNAKIS I. 80 RADU M. 83 SEZGINTURK M.K. 133, 136, 137
PASTORAKOVA A. 158 RADUCANU A. 149, 155 SHEVALYE A. 104
PATRAS A. 187 RAICEVIC S. 116 SHIKOVA E. 158
PAVELKIC V.M. 144 RAICU M. 126 SHUBA I.M. 95
PAVLOVA S. 152 REDDICK A. 75 SIKOLE A. 186
PAZARLIOGLU N.K. 133, 135 REID K.B.M. 114, 163 SIMIONESCU M. 76
PEEVA V. 109 RESAT H. 80 SIMOVA-STOILOVA L. 110, 111
PEJIC S. 101, 139 RESAT M.S. 108 SINICI I. 137, 181
PENKOVA A. 145 RESMI H. 156, 196 SIYAKUS G. 206
PEREIRA-SMITH O.M. 188 RIBARAC-STEPIC N. 78 SLAVESKA N. 129
PEROVIC M. 162 RIBAROV ST. 105 SLAVOV C. 86
PETKANCHIN I.B. 136 RIGHI A. 99 SLYSHENKOV V. 104
PETKOVA D. 112, 119 RIIS B. 216 SMIONESCU M. 76, 83
PETKOVA M. 148 RIPAMONTI E. 129 SOFIC E. 82
PETKOVA M.I. 168, 173 RISTIC S.M. 180 SOGUT E. 211
PETKOVA V. 152 RISTIC-FIRA A. 85, 106, 109, 110 SOGUT S. 184
PETRESCU A.J. 108, 150 RIVNEAC E. 128 SOGUTCU T. 147
PETRESCU S.M. 150, 195 RIVNEAC V. 128 SOZMEN E.Y. 138, 156
PETROVA S. 131, 135 ROBERTS M. 80 SPASOJEVIC-TISMA V.D. 144
PETROVA V. 105 ROSEANU A. 149, 155, 160 SPERDOULI E. 188
PETROVIC I. 85 ROSSO A. 99 SREBREVA L. 98, 123
PETROVIC S.B. 105, 121 ROUMENINA L. 114, 163 STANGA D. 74, 159
PETROVIC T. 167 RUJDIJIC S. 85, 106, 109, 110 STANIC D. 116, 117
PETROVSKA S. 186 RUNDQUIST I. 98 STANKOVIC S. 146, 153, 162
PHILIPS E. 203 RUSKOVSKA T. 186 STANOJKOVIC T. 167
PILLOTON R. 133 RUSSEV G. 74 STEFANOVSKA A.M. 71
PINTZAS A. 80 RUZDIJIC S. 85, 106 STENEVA J. 112, 119
PIPERSKI V.D. 180 • STEPHANOVA E.V. 151, 166
PIROZKOV A. 152 SADI G. 139 STOIAN G. 131
PISKIN A.K. 216 SAGDICOGLU G. 189 STOICA C. 149
Turk J Biochem, 2003; 28(3), 62-224 223 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
STOILKOVA G. 193 TO K. 95 VALENTSIUKEVICH O.I. 125
STOJANOVIC S. 116 TODOROVA B. 131 VALTCHEVA R.V. 166
STOJILJKOVIC V. 101, 139 TODOROVA K. 81 VANGELOVA K. 176
STOJKOVSKI V. 101 TODOROVIC D. 85, 106 VARAGIC V. 124
STOROZHUK L.M. 95 TOKER A. 164 VASIC V. 140
STOYANOVA A. 84, 163 TONCHEVA E. 148 VASSILATIS D.K. 188
STOYANOVA Z. 110, 111 TOPALOGLU E. 209 VELITCHKOVA M. 98, 149
STOYNEV G. 123 TOPCU C. 212 VELKOVA A. 74
STRASSER R.J. 86, 172, 173 TOPOUZOVA T. 151 VERGIDOU C. 97
STUPAREVIC I. 82 TOSHESKA K. 131 VLADIMIROVA D. 109
SUER GOKMEN S. 115, 114 TRAYKOV T. 105 VOINEA M. 83
SUKIASYAN A.R. 185 TRIF M. 155, 161 VOJINOVIC-MILORADIV M. 177
SULEYMAN H. 165, 175 TRIFAN M. 96 VOJNOVSKA Z. 101
SULJEVIC E. 190 TRIFUNOVIC D. 132 VRANIģ MANDUSIģ V. 76, 151
SUNAR B. 115, 116 TROPAK M.B. 181 VUJCIC M. 85, 109, 110
SVETLICIC V. 72 TSACHEVA I. 114, 163, 168 VUKOVA T.I. 189
SVORCAN P. 146 TSENOVA W. 158 VULOVIC M. 78
SWAROOP A. 75 TSOLKAS G.P. 87 VUNKOVA-RADEVA R. 144
SYMEONIDIS L. 98 TUDORIE M. 194 •
SYROS T.D. 99 TULI A. 140, 150, 165 WEBSTER V. 204
SZEDLACSEK S.E. 74, 159 TULUCE Y. 210 WILEY H.S. 80, 108
• TUMANYAN L.R. 93 WOLSTENHOLME A. 85
TABAKCIOGLU K. 107 TUNCER O. 208 •
TAMER L. 156 TUNCKALE A. 100 YAKOVENKO O. 94
TAMER S. 126 TURAN A. 78 YALCIN A. 138, 156
TANDOGAN B. 183, 211 TURAN B. 207 YALCIN A.S. 158
TANELI F. 207, 208, 209 TURK M. 201, 206 YALIN E.A. 174
TANEVA S. 136, 172 TURKALP I. 142 YALIN S. 129
TANIC M. 190 TURKOZ Y. 187 YANIKKAYA-DEMIREL G. 184
TANIC N. 132 TUTESKA J. 101 YARDIMCI T. 183, 184
TANRIVERDI K. 140 TUYLU T. 130 YARGICOGLU P. 212, 213
TANYALCIN O. 204 TUZUN C. 209 YASA I. 135, 137
TANYALCIN T. 89, 186, 203, 204 TZONTCHEVA A. 95, 114 YAVUZKIR M. 205
TARHAN L. 161, 171 TZVETKOV M. 94 YENISEY C. 193
TASIC J.S. 180 • YERER M.B. 191
TASIC V. 129 UCAKTURK L. 142 YIGIT R. 126
TASKIRAN D. 186 UCAR F. 119 YILDIRIM I. 157
TCHORBADJIEVA M. 163, 167, 168, UCAR G. 209 YILDIRIM N. 100
173 UCHIBA M. 155, 175 YILDIRIM O.S. 167
TEKE M. 134 UGURLU S. 200 YILDIZ C. 214
TEKELIOGLU Y. 119 ULMAN C. 207, 208, 209 YILDIZ G. 82
TEKIN H. 100 ULUSOY G. 178 YILDIZ O. 191, 192
TEKSEN M. 113 ULUSU N.N. 183, 207, 211 YILDIZ R. 114
TELEFONCU A. 72, 117, 133, 134, ULUTAS F. 130 YILMAZ C. 92, 134, 153, 154
137, 138, 153, 154, 170 UNGUREANU D. 102, 103 YILMAZ D. 208
TEMEL I. 133, 159, 182 UNLU Z. 209 YILMAZ F. 113, 118
TENCHOV B.G. 160 UNLUKURT I. 104 YILMAZ H. 93
TEPARIC R. 82 USLU A. 211 YILMAZ O. 72
TERZIC N.A. 109, 121 USTA D. 164 YILMAZ R. 133, 182
TETIK S. 183 UTKU H. 102 YILMAZ Z. 202
TEZCAN E.F. 183 UYANIK B.S. 207, 208, 209 YOKUS B. 79, 141, 146, 147, 177
TIKIZ C. 209 UYSAL O. 100 YOLOGLU S. 187
TIKIZ H. 209 UZBAY I.T. 73, 194 YONOVA P. 193
TIMUR S. 133, 134 • YORDAM N. 88
TIRCOL M. 156 VACARU A. 74, 159 YORDANOV I. 86
Turk J Biochem, 2003; 28(3), 62-224 224 http://www.TurkJBiochem.com
13th Balkan Biochemical Biophysical Days & Meeting on Metabolic Disorders’ Programme & Abstracts
YORDANOVA-ZLATORAVA R. YUZGEC V. 204, 205 ZIHNIOGLU F. 72, 134, 135, 137,
171, 181 • 138, 153, 154
YUCEL O. 165, 175 ZAFIRIADIS H. 99 ZIZAK Z. 167
YUKSEL Mehtap 155, 175, 196 ZAHARIEVA I. 86, 172 ZLATAROVA A. 114, 163
YUKSEL Meral 158 ZAKULA Z. 78 ZLATEVA G. 109
YUPSANI A. 187 ZEINALOV Y. 145 ZORKO M. 79
YUPSANIS T.A. 97, 98, 99, 187 ZEREN T. 195 ZORLU F. 73
YURTSEVER E. 169 ZEYNIYEV A. 189 ZUTIC V. 72
YURTTAS B. 172 ZEYTINOGLU H. 140 ZVERINSKY I. 104
YUZBASIOGLU S. 104 ZHUKOV O.D. 94, 95
Turk J Biochem, 2003; 28(3), 62-224 225 http://www.TurkJBiochem.com