Symposium I Physiological phenotype of genetically modified by xiuliliaofz

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									                                                   CONTENTS




                                                                                                           page

Welcome to Oulu ............................................................................................3

General information ........................................................................................4

Programme ......................................................................................................9

Abstracts .......................................................................................................33

Author index ...............................................................................................142

Campus map................................................................................................148

Sponsors ......................................................................................................149

Exhibitors ....................................................................................................150




                                                            1
                                       General information


                                      Welcome to Oulu

On the behalf of the Scandinavian Physiological Society (SPS) and the local organizing committee,
it is my great honour and pleasure to welcome our members and guests to the Scandinavian
Physiological Society Meeting in Oulu.

The organizing committee invited three state-of-the-art lectures by Greg Stuart, Kari Alitalo and
Mart Saarma, selected 19 specialized symposia from an open call in fall last year and 4 oral and 11
poster sessions from submitted abstracts. The programme is made up of 3 parallel tracks except
Sunday. On Sunday we will have in addition a special panel discussion forum „Physiology 2008 –
future directions and threats‟ with representatives from industry, academia and physiological
associations (SPS and FEPS). The meeting will have almost 300 participants from 18 countries.

The abstract committee selected from the 96 submitted abstracts 18 for oral and 78 for poster
presentations. Posters will be presented in small groups chaired by distinguished members of the
Society. From the presenters (oral and poster each) three "Young Investigator Awards" will be
chosen by the Chairpersons who will act as the jury.

Our meeting is part of 50th Anniversary Celebrations of the University of Oulu and will be opened
by the Rector of Oulu University Lauri Lajunen and Minister Suvi Lindén. The University of Oulu is
one of the largest universities in Finland providing us with the academic environment to foster
exchange of ideas, collaboration and friendship.

The City of Oulu cordially invites all attendees of the meeting to attend the Welcome Reception at
the City Hall on Friday. The Congress Dinner will be held at one of the most beautiful cultural
heritage sites of Oulu, the Maikkulan Kartano manor house on Saturday.

We hope that you will enjoy the meeting and your stay in Oulu and the North of Finland!


Cordially,


Karl-Heinz Herzig

Chairman of the Organizing Committee




                                                  3
                                          General information


                                        General information

Local organizing committee

Karl-Heinz Herzig, meeting president
Esa Hohtola
Johanna Issakainen
Juhani Leppäluoto
Satu Mänttäri
Petteri Nieminen
Anna-Kaisa Purhonen
Seppo Saarela
Hannele Savela
Olli Vuolteenaho


Language

All sessions will be held in English.


Meeting Rooms

The SPS 2008 Annual Meeting takes place in the Saalasti-hall and nearby auditoria, located right at
the heart of the university main building at Linnanmaa campus area close to the main entrance.


Speaker preview room

Speakers are required to deliver their presentation to the speaker preparation room during the
morning for afternoon presentations and during the afternoon for next-morning presentations. The
presentations should be delivered on a memory stick or CD. Only PowerPoint presentations are
accepted and only single-projector presentations should be delivered.


Transportation to and from congress venue

Bus transportation

The distance between the city centre and the University of Oulu in Linnanmaa is ca. 5.5 km/10–15
minutes by bus. There will be a free transportation available from the congress hotels (Radisson
SAS, Scandic, Arina, Cumulus, Turisti) to the congress venue in the morning and back to hotels in
the afternoon. The schedule for bus transportation can be found in the congress bag and also at the


                                                  4
                                        General information

hotels. The bus can only stop for a couple of minutes at each congress hotel. In case your
accommodation is not in any of the official congress hotels, please arrive to the nearest congress
hotel for organized transportation or use the public bus transportation in Oulu.

Public bus transportation in Oulu

If you miss the congress transportation, you can use public buses. Numbers 4, 6, 7, 19 operate from
the city (e.g. Toripakka bus stop) to the Linnanmaa campus. The bus stop is on the east side of the
campus, from where congress signs will guide you to the meeting place.

A single ticket in the area of the city costs EUR 2.90.

Taxi

Dial 10041 from normal phones, 0600-30081 from mobile phones, +358 600 30081 from abroad.
For Airport taxi, dial 0600-30084.


Meeting secretariat

The meeting secretariat will be located in the lobby of the main building near to the main entrance.
The desk will be open at all times during the scientific sessions.

At the congress site (and many parts of the city), you can easily stay on-line with the free
panOULU (public access network OULU) network that provides in its coverage area wireless
broadband Internet access to everybody.


Free oral presentations

Free oral presentation sessions take place on Friday 15 starting 16.30 and Saturday 16 starting at
13:00 (three parallel sessions). Oral presentations are accepted only as PowerPoint presentations.
Each presentation has been allocated 15 minutes in the programme (10 minutes for presentations + 5
minutes for discussion). The chairman should make sure that the time limits are followed.


Poster sessions

The poster area is located in the area between the Saalasti-hall and nearby auditoria. Posters are
combined into special groups for guided sessions. Each poster will be on display for duration of the
meeting. The guided poster session is on Friday 15th (13.00–14.00) during the longer break after
lunch time. The presenters are kindly asked to be at their posters at that time. Each presenter should
give a short oral presentation (about 5 minutes) about his/her poster, which is followed by a short
general discussion. The presenters are to remain at their poster sites during these guided poster
sessions.


                                                   5
                                        General information

Poster chairs should meet in room HR144 at 16.00 after the sessions to select the best three posters
from presenters under 35 years for the Young Scientist Award .


Credit points for students

Participation in the meeting will be approximately 1 ECTS and participation with a poster/oral
presentation a total of 2 ECTS. The certificate of participation should be presented to the individual
faculty the students belongs to. The final decision on the credit will by the responsible faculty only.


Lunch and coffee breaks

A take-away lunch will be served in the Saalasti-hall. Together with the name badge, participants
will receive lunch and coffee tickets for the congress days. The coffee will be served in the lobby
outside the auditoria. See map at the end of this booklet.


Commercial exhibition

There will be a commercial exhibition in the lobby of the main building across the Saalasti-hall. The
exhibition will be open on Friday and Saturday at 8:30-17:00 and on Sunday at 8:00-12:00. The list
of exhibitors can be found at the end this booklet. We kindly ask the participants to visit the
exhibition.


Tourist information

Oulu City Tourist Office
Uusikatu 26, P.O.Box 32
FIN-90015 Oulun Kaupunki
Open: Mon-Fri:9.00-16.00
Tel: +358 8 5584 1330
Fax: +358 8 5584 2099
Website: www.oulutourism.fi
Email: touristinfo@ouka.fi


Social programme

Welcome Reception

The City of Oulu kindly invites all participants to a Welcome Reception on Friday evening. The
reception takes place at the City Hall of Oulu, which is located right at the city centre of Oulu,
within a walking distance from the hotels. Buses leave straight from the congress venue to the City

                                                   6
                                        General information

Hall at 18.30. The reception starts at 19.00. Cocktail-type finger food and refreshments will be
served at the reception. The Welcome Reception is included in the registration fee.

Congress Dinner and Social Programme

The Congress Dinner is held at one of the most beautiful cultural heritage sites of Oulu, the
Maikkulan Kartano manor house on Saturday, August 16 at 19.00. Maikkulan Kartano is located
only a 15-minute drive from the City centre of Oulu. Bus transportation between the congress hotels
and Maikkulan Kartano will be arranged. During the evening at Maikkulan Kartano, you are offered
a unique chance to experience the Finnish sauna. Separate sauna for males and females will be
available. Towels are provided on site. The social program also includes music and dancing on the
outside patio. Participation requires the purchase of a special dinner card.




                                                  7
                                         Friday August 15


                                          Programme

   The presenting author are listed in the program. See the abstracts for complete lists of authors.


07.30 – 08.30 Registration, Linnanmaa Campus, main building

08.30 – 09.00 Opening words (Saalasti hall)

08.30      Karl-Heinz Herzig, President of the Organizing Committee

08.35      Lauri Lajunen, Rector of the University of Oulu

08.45      Suvi Lindén, Minister of Communications, M.P.


09.00 – 10.00 State-of- the-art Lectures (Saalasti hall)
              Chair: Taina Pihlajaniemi

09.00      The action potential
           Greg Stuart, Australia
09.30      Molecular regulation of angiogenesis and lymphangiogenesis
           Kari Alitalo, Finland


10.00 – 10.30 Coffee break

Symposium 01 (Auditorium “Tigerstedt”)

10.30 – 12.30 Physiological phenotype of genetically modified animals
              Chairs: Rolf Reed, Helge Wiig

10.30      Physiological monitoring of microcirculation in genetically modified animals
           (S0101)
           Fitz-Roy Curry, USA
10.54      Physiological function of cytoskeleton and motor proteins in muscle studied in
           transgenic mice and zebrafish larvae (S0102)
           Anders Arner, Sweden
11.18      Structural and regulatory roles of conserved collagens (S0103)
           Taina Pihlajaniemi, Finland

                                                  9
                                    Friday August 15


11.42    Mouse models for skin development and skin diseases (S0104)
         Cord Brakebusch, Denmark
12.06    Use of imaging techniques in analysis of mouse phenotypes (S0105)
         Frits Thorsen, Norway


Symposium 02 (Auditorium “Granit”)

10.30 – 12.30 Physical activity and health - Human physiology in changing
              thermal environments
              Chairs: Tiina Mäkinen, Juhani Leppäluoto

10.30    Climate indices for human performance and health in outside conditions (S0201)
         Ken C. Parsons, UK
11.00    Heat, hydration and exercise (S0202)
         Ron Maughan, UK
11.30    Cardiovascular regulation in cold (S0203)
         Matti Mäntysaari, Finland
12.00    Neuromuscular performance in cold (S0204)
         Juha Oksa, Finland


Symposium 03 (Auditorium “Donner”)

10.30 – 12.30 Neuronal excitability: synaptic vs. extrasynaptic influence (MTP-
              symposium)
              Chairs: Michael Druzin, Staffan Johansson

10.30    Voltage-sensor modulation in K channels regulates neuronal excitability (S0301)
         Fredrik Elinder, Sweden
10.54    Functional role of tonic GABA in the hippocampus (S0302)
         Matthew Walker, UK
11.18    P2X mediated transmission in neuronal-glial networks (S0303)
         Alexei Verkhratsky, UK
11.42    Tonically prebound extrasynaptic NMDA receptors (S0304)
         Alexey Semyanov, Russia
12.06    Chloride concentration changes determine current time course and mediate
         interaction between GABA receptors and glycine receptors (S0305)
         Staffan Johansson, Sweden


                                           10
                                        Friday August 15


12.30 – 13.00 Lunch

13.00 – 14.00 Poster sessions 01-11

Session 01       Cardiovascular & pulmonary physiology
                 Chair: Tobias Wang

P01
Inflammation and electrical instability in chronic myocardial infarction
Ioana Mozos, Romania

P02
Isolated factors identified in the effluent of preconditioned hearts offers cardioprotection to
donor hearts
Lars Breivik, Norway

P03
Effects of purine compounds on bioelectrical activity of hibernating ground squirrel heart
Vladislav Kuzmin, Russia

P04
Chronic administration of serotonin transporter inhibitor (fluoxetine) decreases
monocrotaline-induced pulmonary hypertension in rats
Valeriya Kozhevnikova, Russia

P05
Gonadectomy attenuates hypoxia-induced pulmonary hypertension in male rats
Olga Kudryavtseva, Russia

P06
Endothelin converting enzyme inhibitor decreases endothelin-1 plasma level and increases NO
production with urine in rats with hypoxia induced pulmonary arterial hypertension
Simonova AI, Russia

P07
Characterization of ecstasy (MDMA)-induced blood pressure, heart rate and respiratory rate
changes in anaesthetized female rats
Rungrudee Srisawat, Thailand




                                                11
                                       Friday August 15


Session 02      Cardiovascular physiology – peripheral mechanisms,
                Chair: Christian Alkjaer

P08
Vessel permeability of a macromolecular contrast agent in mouse muscle using Magnetic
Resonance Imaging (MRI)
Guro Løkka, Norway

P09
Role of cyclooxygenase-2 for lipopolysaccharide-mediated vasorelaxation in vitro and blood
pressure decrease in vivo
Mette Staehr, Denmark

P10
Hypotensive effect of oxatriazolium-5-olate derivative in spontaneous hypertensive rats (SHR)
during chronic introduction
Marina Artemieva, Russia

P11
The role of mitogen-activated protein kinase in the contraction of rat saphena artery
Dina Gainullina, Russia

P12
The role of Rho-kinase in adrenergic vasopressor response decreases during postnatal
development
Nadezda Tarasova, Russia


Session 03      Cells, receptors, channels, membranes
                Chair: Jyrki Kukkonen

P13
Biochemical characterization and comparison of N-terminal Gly16Arg variants of the 2-
adrenergic receptor
Minna Lilja, Finland

P14
Regulation of cardiac activity by Kir2.x inward rectifier potassium channels in fish
Vesa Paajanen, Finland

P15
Electrophysiological properties fish Kir2.x channels expressed in mammalian cell lines
Tomi Pekurinen, Finland




                                              12
                                       Friday August 15

P16
The effect of osmotic shock on the early mouse embryo volume
Maria Pogorelova, Russia

P17
The antimicrobial peptide plantaricin A permeabilizes liver and kidney cells
Kristin Andersland, Norway

P18
Both normal and cancerous lymphocytes and neurons are permeabilized by plantaricin A, a
peptide produced by Lactobacillus plantarum
Sverre L. Sand, Norway

P19
Determination of cell viability of mesenchymal stem cells by analyzing mitochondrial function.
Mika Pietilä, Finland


Session 04      Endocrinology and metabolism
                Chair: Petteri Nieminen

P20
Role of ACBP in food intake and hypothalamic gene expression in transgenic rats
Anne Huotari, Finland

P21
Regulation of visfatin expression by feeding status in mice
Anne Huotari, Finland

P22
Increased heat production and upregulation of PPARs and UCP2 mRNA in WAT in mice
overexpressing human prepro-orexin
Kari Mäkelä, Finland

P23
Evaluation of association between leptin levels and leptin gene polymorphism in obese females
Nilsel Okudan, Turkey

P24
Effects of reported previous physical activity on body composition and its changes during
military service
Tiina Mäkinen, Finland

P25
Effects of coenzyme Q10 supplementation on plasma adiponectin, interleukin-6 and tumor
necrosis factor-alpha levels in males
Muaz Belviranlı, Turkey

                                               13
                                        Friday August 15

P26
Identification of possible ovarian cancer specific biomarkers in the tumour interstitial fluid
Hanne Hox, Norway

P27
The estimation of milk intake and energy expenditure of reindeer calves by the doubly-labelled
water method
Päivi Soppela, Finland


Session 05       Muscle and exercise physiology I
                 Chair: Juha Oksa

P28
The use of a cooling vest during exercise in warm condition delays the appearance of fatigue
and improves cycling performance
Benoit Dugue, France

P29
Effects of menstrual cycle, oral contraception, and training on exercise-induced changes in
circulating DHEA-sulphate and testosterone in young women
Benoit Dugue, France

P30
Soldiers' performance during a two-week field exercise in winter
Sirkka Rissanen, Finland

P31
The swimming performance of brown trout and whitefish: the effects of exercise on Ca2+
handling and oxidative capacity of swimming muscles
Katja Anttila, Finland

P32
-Lipoic acid attenuates exercise-induced oxidative stress without impairing exercise capacity
Susanna Kinnunen, Finland


Session 06       Muscle and exercise physiology II
                 Chair: Jan Henriksson

P33
Endurance training exerts differential effects on diaphragm and gastrocnemius muscle feed
arteries in the rat
Anna Borzykh, Russia




                                               14
                                       Friday August 15

P34
The reproducibility of knee cartilage magnetic resonance imaging with dGEMRIC method
Johanna Issakainen, Finland

P35
Collagen XIII is a postsynaptic, multifunctional neuromuscular junction protein
Anne Latvanlehto, Finland

P36
Effects of dexamethasone and exercise on rat myocardial collagen synthesis
Timo Takala, Finland

P37
Time-course of exercise needed for bone changes in a 12-month intervention study
Riikka Heikkinen, Finland

P38
Impact exercise alters long-term bone turnover in a dose-dependent manner
Aki Vainionpää, Finland

P39
Contractile defects and protein nitration in slow-twitch muscle of mice with collagen-induced
arthritis
Takashi Yamada, Sweden


Session 07      Gastrointestinal physiology
                Chair: Gunnar Flemström

P40
Oral bacteria regulate gastric mucosal defense via bioactivation of dietary nitrate
Joel Petersson, Sweden

P41
Influence of short glyprolines with arginine on C-end on gastric mucosa homeostasis in
ethanol-induced ulcerogenesis
Alexandra Puchkova, Russia

P42
Peripheral and central mechanisms of gastric mucosa homeostasis and glyprolines
Galina Samonina, Russia

P43
GFRα2 knockout mouse as a model for studying autonomic regulation of gastric acid and
ghrelin secretion
Jussi Kupari, Finland


                                               15
                                      Friday August 15

P44
Defective jejunal and colonic salt absorption and altered Na+/H+ exchanger 3 (NHE3) activity
in NHE regulatory factor 1 (NHERF1) adaptor protein deficient mice
Mingmin Chen, Germany

P45
Dual role of the Na+/H+ exchanger isoform 3 for PEPT1-mediated H+/dipeptide cotransport in
native murine intestine
Mingmin Chen, Germany

P46
Differential role for the PDZ proteins NHERF1, NHERF2 and PDZK1 in the regulation of
CFTR-mediated intestinal anion secretion in vivo
Ursula Seidler, Germany

P47
Key role of CFTR for all modes of intestinal HCO3- secretion
Ursula Seidler, Germany

P48
Chronic PD induces remodeling of the peritoneal membrane and local secretion of TGF- and
VEGF in rats
Solfrid Olsen, Norway


Session 08      Neurophysiology I
                Chair: Bryndis Birnir

P49
Interspike intervals of human motor units during voluntary recruitment
Alexander Meigal, Russia

P50
Coenzyme Q10 and alpha lipoic acid supplementation to diabetic rats: Conduction velocity
distributions
Hakkı Gökbel, Turkey

P51
EEG burst suppression recorded from depth electrodes during anesthesia
Ville Jäntti, Finland

P52
TIRF study of P2X3 receptors trafficking in cultured neurons
Evgeny Pryazhnikov, Finland




                                             16
                                      Friday August 15

P53
Interactions with calnexin and ER calcium pump SERCA2b regulate human delta opioid
receptor maturation
Jussi Tuusa, Finland

P54
Taurine reduces ethanol-induced caspase-3 activation in the developing cerebellum
Andrey Taranukhin, Finland

P55
Differential roles of L-type Ca2+ channels for evoked and spontaneous transmitter release onto
rat preoptic neurons
Evgenya Malinina, Sweden

P56
Postponed effects of chronical neonatal arginine-vasopressin structural analogue (Ас-D-Met-
Pro-Arg-Gly-NH2) administration on learning processes in white rats
Polina Kim, Russia

P57
Exogenous heat shock protein 70 affects sleep and thermoregulation after sleep deprivation in
pigeons
Ksenia Lapshina, Russia


Session 09      Neurophysiology II
                Chair: Esa Korpi

P58
Millisecond substance exposure on HEK-293 cells expressing several GABA-A receptor
subtypes using patch-clamp in DynaflowTM system
Samira Gouissem, Sweden

P59
Insulin modulation of GABA-A receptors-mediated inhibition in rat hippocampus
Zhe Jin, Sweden

P60
AA29504 – a modulator of extrasynaptic GABA-A receptors
Kirsten Hoestgaard-Jensen, Denmark

P61
GABA input into orexin neurons is involved in proper maintenance of vigilance state
Taizo Matsuki, Japan




                                              17
                                        Friday August 15

P62
Neurosteroids 3,20 (R/S)-pregnandiols decrease offset rate of the GABA-site activation at the
recombinant GABAA receptor
Ming-De Wang, Sweden

P63
Milk Sugar Test; social competition model for study of direct effect of GABAA receptor active
compounds
Magnus Bengtsson, Sweden

P64
Extrasynaptic-like GABA channels expressed in lymphocytes
Suresh Kumar Mendu, Sweden


Session 10       Renal and comparative physiology
                 Chair: Ole Skøtt

P65
Plasmin stimulates the epithelial sodium channel via interaction with prostasin
Per Svenningsen, Denmark

P66
The significance of prostasin for tight epithelial formation
Mette Steensgaard, Denmark

P67
Lithium impairs kidney development and inhibits glycogen synthase kinase-3β activity
Gitte Kjaersgaard, Denmark

P68
Dihydropyridine and ryanodine receptors in avian skin – what for?
Liisa M. Peltonen, Finland

P69
Hypoxia and adenosine in the epaulette shark
Kalle Rytkönen, Finland

P70
Influence of acute hypoxia on antioxidant system of non-pregnant and pregnant rats
Lidia Trofimova, Russia

P71
Regulation of blood pressure during head movement in the anaesthetized giraffe
Emil Brøndum, Denmark



                                                18
                                        Friday August 15


Session 11       Other topics
                 Chair: Peter Bie

P72
Effect of prolyl-glycyl-proline (PGP) peptide on mast cells activity in vitro
Nadezhda Bondarenko, Russia

P73
Photoplethysmographic measurement and pulse wave analysis
Matti Huotari, Finland

P74
Smoking habits among university students in Tartu, Estonia and Oulu, Finland
Peet-Henn Kingisepp, Estonia

P75
Enhanced cold-induced vasoconstriction (Raynaud’s phenomenon) is the sign of disadaptation
to cold
Ludmila Gerasimova, Russia

P76
Neuronal NOS contributes to long-term facilitation of neck muscle nociception in mice
Dejan Ristic, Denmark

P77
Gender differences in cooling-induced changes in muscular activity
Erja Sormunen, Finland

P78
Performing cognitive task increases oxygen consumption during low intensity work at
thermally neutral and cold environment
Nicolas Troubat, France


Symposium 04 (Auditorium “Tigerstedt”)

14.00 – 16.00 Neurotransmission and neuromodulation I
              Chair: Esa Korpi


14.00      GABA-A receptors mediating tonic inhibition in the neocortex (S0401)
           Kimmo Jensen, Denmark
14.30      GABA mediated inhibition in rat hippocampus is modulated by insulin (S0402)
           Bryndis Birnir, Sweden


                                                19
                                     Friday August 15


15.00    Glutamatergic adaptation of dopamine neurons to single doses of benzodiazepines
         (S0403)
         Anne Heikkinen/Esa Korpi, Finland


Symposium 05 (Auditorium “Granit”)

14.00 – 16.00 Modern imaging technologies in the study of human physiology
              Chair: Kari Kalliokoski

14.00    Use of Doppler ultrasound and coronary CT to study coronary circulation and
         vascular endothelial function in man (S0501)
         Jaakko Hartiala, Finland
14.30    Evolving magnetic resonance imaging (MRI) and spectroscopy (MRS) techniques in
         the study of human physiology (S0502)
         Ronald Borra, Finland
15.00    Molecular imaging of metabolism using positron emission tomography (PET)
         (S0503)
         Pirjo Nuutila, Finland
15.30    Microvascular blood flow and oxygenation measured with near-infrared
         spectroscopy (S0504)
         Robert Boushel, Denmark


Symposium 06 (Auditorium “Donner”)

14.00-16.00   Gastrointestinal barrier and transport functions in health and
              disease
              Chairs: Ursula Seidler, Gunnar Flemström

14.00    New anion transport proteins in the upper GI tract (S0601)
         Manoocher Soleimani, Germany
14.24    Intragastric nitric oxide and gastric mucosal defense (S0602)
         Joel Petersson, Sweden
14.48    Congenital chloride diarrhea – a Finnish disease serves as a paradigm for
         electroneutral NaCl absorption (S0603)
         Georg Lamprecht, Germany
15.12    Regulatory aspects of the paracellular barrier: special role of claudin-2 and -8
         (S0604)
         Salah Amasheh, Germany


                                             20
                                     Friday August 15


15.36    The role of the NHERF family of PDZ-scaffolding proteins in the regulation of salt
         and water transport: lessons learned from knockout mice (S0605)
         Ursula Seidler, Germany


16.00 – 16.30 Coffee break

Symposium 07 (Auditorium “Tigerstedt”)

16.30 – 18.30 Neurotransmission and neuromodulation II
              Chair: Eric Hanse

16.30    Monocarboxylate transporters at brain synapses (S0701)
         Linda H Bergersen, Norway
16.54    Glutamate release from astrocytes (S0702)
         Vidar Gundersen, Norway
17.18    Presynaptic mechanisms of plasticity in the developing hippocampus (S0703)
         Sari Lauri, Finland
17.42    Novel roles of kainate receptors in hippocampal development (S0704)
         Tomi Taira, Finland
18.06    Developmental aspects of short- and long-term plasticity in the hippocampus
         (S0705)
         Eric Hanse


Symposium 08 (Auditorium “Granit”)

16.30 – 18.30 Physical activity and health - Cardiovascular diseases, diabetes,
              and osteoporosis
              Chairs: Timo Takala, Juhani Leppäluoto

16.30    Individual differences in responsiveness to regular physical activity (S0801)
         Tuomo Rankinen, Finland
17.00    Impact of intense exercise on BMD, fall risk and 10y-CHD-risk in elderly females.
         Results of the Senior Fitness and Prevention Study (SEFIP) (S0802)
         Wolfgang Kemmler, Germany
17.30    Obesity: Fat oxidation and insulin resistance (S0803)
         Michelle Venables, UK




                                             21
                                     Friday August 15


18.00    Aspects on physical activity measurements (S0804)
         Timo Jämsä, Finland


Free oral presentations 01 (Auditorium “Donner”)

16.30 – 18.30 Gastrointestinal physiology
              Chair: Ursula Seidler

16.30    Luminal glucose is a prerequisite for a stimulation of duodenal alkaline secretion
         by orexin-A (F0101)
         Magnus Bengtsson, Sweden
17.15    Apelin induced stimulation of duodenal bicarbonate secretion (F0102)
         Gunnar Flemström, Sweden
17.30    LPS and PGN stimulate mucus secretion in colon (F0103)
         Mia Phillipson, Sweden
17.45    Genetic ablation of the K+ channels Kir 4.1 and KCNQ1 has opposing effects on
         acid secretory rates in gastric mucosa of weanling mice (F0104)
         Penghong Song, Germany
18.00    Angiotensin II regulates the mRNA expression of enzymes involved in arginine
         metabolism in isolated renal resistance vessels through the AT1 receptor (F0105)
         Michael Hultström, Norway
18.15    Intact angiotensin II signalling in the postnatal period is necessary for normal
         development of the renal microcirculation (F0106)
         Kirsten Madsen, Denmark




Transportation to City hall/hotels – buses leave at 18.30

Welcome reception, City Hall of Oulu, 19.00 – 20.30




                                             22
                                    Saturday August 16


Symposium 09 (Auditorium “Granit”)

08.00 – 10.00 ATP and adenosine - receptors and roles
              Chair: Bertil Fredholm

08.00    Adenosine and ATP - receptors and formation
         Bertil Fredholm, Sweden
08.30    Extracellular adenosine generation and signaling in cardiovascular, pulmonary and
         renal injury (S0902)
         Holger Eltzschig, USA
09.00    Adenosine, ATP and the regulation of sleep/wakefulness (S0903)
         Tarja Stenberg, Finland
09.30    Knocking out the adenosine A1 receptor increases lipolysis and insulin release
         (S0904)
         Bertil Fredholm, Sweden


Symposium 10 (Auditorium “Donner”)

08.00 – 10.00 Senses at the threshold
              Chair: Kristian Donner

08.00    Vision at extreme (S1001)
         Viktor Govardovskii, Russia
08.30    Colour vision limits in dim light (S1002)
         Almut Kelber, Sweden
09.00    Mammalian hearing sensitivity in air, and in water (S1003)
         Sirpa Nummela, Finland
09.30    Trade-offs in visual detection (S1004)
         Kristian Donner, Finland




                                             23
                                   Saturday August 16


Symposium 11 (Auditorium “Tigerstedt”)

08.00 – 10.00 Long-term blood pressure regulation in normal man:
              New aspects of neuro-humoral control
              Chair: Peter Bie

08.00    Proximal sodium reabsorption: a role in the regulation of blood pressure? (S1101)
         Michel Burnier, Switzerland
08.30    Sodium intake, renal sodium reabsorption, and genetic predisposition in long term
         blood pressure control (S1102)
         Olle Melander, Sweden
09.00    Long-term baroreflex and short-term renin-angiotensin regulation of blood
         pressure in humans (S1103)
         Peter Norsk, Denmark
09.30    Blood volume, blood pressure, and total body sodium: internal signaling and output
         control (S1104)
         Peter Bie, Denmark


10.00 – 10.30 Coffee break

Symposium 12 (Auditorium “Granit”)

10.30 – 12.00 The effects of sleep loss on performance and human health
              Chair: Tarja Stenberg

10.30    Sleep loss and accident risk (S1201)
         Tor-Börn Åkerstedt, Sweden
10.53    Sleep loss and immune function (S1202)
         Tarja Stenberg, Finland
11.15    Cognitive performance during sleep loss: What are we measuring? (S1203)
         Marja-Leena Haavisto, Finland
11.38    Sleep, mood and genes (S1204)
         Tiina Paunio, Finland




                                            24
                                      Saturday August 16


Symposium 13 (Auditorium “Tigerstedt”)

10.30 – 12.00 Oxygen-dependent regulation of gene expression
              Chairs: Minna Vainio, Mikko Nikinmaa

10.30    Molecular imaging of cellular oxygen sensing (S1301)
         Joachim Fandrey, Germany
11.00    Title TBA (S1302)
         Lorenz Poellinger, Finland
11.30    Evolution of hypoxia-inducible factor in fish (S1303)
         Kalle Rytkönen, Finland


Symposium 14 (Auditorium “Donner”)

10.30 – 12.00 The role of nitric oxide in the development of hypertension
              Chair: Erik Persson

10.30    Role of NO deficiency in salt-sensitive hypertension (S1401)
         Mattias Carlström, Sweden
10.53    Mechanisms of NO release in angiotensin II induced hypertension in rats (S1402)
         Bjarne Iversen, Norway
11.15    The role of the nitrate-nitrite-nitric oxide pathway in vascular control (S1403)
         Joel Petersson, Sweden
11.38    Cyclic AMP-mediated activation of eNOS by vasodilator agonists; role in blood
         pressure regulation (S1404)
         Boye Jensen, Denmark


12.00 – 13.00 Lunch

12.00 – 13.00 SPS General Assembly (Saalasti Hall)




                                             25
                                    Saturday August 16


Free oral presentations 02 (Auditorium “Tigerstedt”)

13.00 – 14.00 Cardiovascular physiology
              Chair: Uwe Pohl

13.00    Optical mapping of the rabbit sinoatrial node under cholinergic and adrenergic
         influence (F0201)
         Denis Abramochkin, Russia
13.15    Mitochondrial ROS triggered by β-adrenergic stress contributes to cardiac
         inotropy in wild-type but not ob/ob mice (F0202)
         Daniel Andersson, Sweden
13.30    Postnatal maturation of arterial smooth muscle (ASM): the impact of sympathetic
         innervations (F0203)
         Olga Tarasova, Russia
13.45    The role of Rho-kinase in regulation of Ca2+-sensitivity of saphenous artery
         contraction in newborn and adults rats (F0204)
         Stepan Mochalov, Russia


Free oral presentations 03 (Auditorium “Granit”)

13.00 – 14.00 Neurophysiology
              Chair: Bertil Fredholm

13.00    Purinergic P2X and P2Y receptors – Opponents in neck muscle nociceptive
         processing (F0301)
         Jens Ellrich, Denmark
13.15    Spatial organization of nociceptive long-term depression in man (F0302)
         Kerstin Jung, Denmark
13.30    ATP and nitric oxide (NO) interfere in microglial responses to spinal cord injury in
         vivo in mice (F0303)
         Eike D. Schomburg, Germany
13.45    Pancreatic islet revascularisation in the intramuscular transplantation site studied
         in vivo (F0304)
         Gustaf Christoffersson, Sweden




                                             26
                                   Saturday August 16


Free oral presentations 04 (Auditorium “Donner”)

13.00 – 14.00 Muscle & exercise physiology
              Chair: Håkan Westerblad

13.00    The effect of adenosine, hypoxia and exercise on local skeletal muscle blood flow in
         humans (F0401)
         Ilkka Heinonen, Finland
13.15    Cardiac output, common and deoxygenated hemoglobin in working muscle during
         incremental bicycle test in subjects of various aerobic performance levels (F0402)
         Daniil Popov, Russia
13.30    Effects of repeated bout exercise on inhibition of extracellular matrix degradation
         on rat muscle (F0403)
         Timo Takala, Finland
13.45    Ketone bodies inhibit insulin-mediated glucose transport in mouse skeletal muscle
         (F0404)
         Abram Katz, Sweden


14.00 – 14.30 State-of- the-art Lecture (Saalasti hall)
              Chair: TBA

14.00    Neurotrophic factors and their receptors in development and disease
         Mart Saarma, Finland


14.30 – 15.00 Coffee break

Symposium 15 (Auditorium “Tigerstedt”)

15.00 – 17.00 Calcium signaling in heart and muscle
              Chairs: Satu Mänttäri, Niels Ørtenblad

15.00    Cardiac development, dysrhythmia, inotropy and hypertrophy; the many possible
         roles of inositol 1,4,5-trisphosphate in the heart (S1501)
         Martin Bootman, UK
15.30    Contribution of sarcoplasmic reticulum to excitation-contraction coupling of the
         fish heart (S1502)
         Matti Vornanen, Finland



                                            27
                                   Saturday August 16


16.00    Changes in skeletal muscle calcium handling during fatigue and recovery (S1503)
         Håkan Westerblad, Sweden
16.30    Metabolic modulation of skeletal muscle Ca2+ handling and excitability (S1504)
         Niels Ørtenblad, Denmark


Symposium 16 (Auditorium “Granit”)

15.00 – 17.00 Ecophysiology and energetics
              Chairs: Seppo Saarela, Esa Hohtola

15.00    Metabolic senescence in a small passerine bird, the zebra finch (S1601)
         Claus Bech, Norway
15.30    Coping with oxygen shortage - physiological adaptations of the deep-diving hooded
         seal (S1602)
         Lars Folkow, Norway
16.00    Comparative physiology of fatty acid mobilization and hepatic lipidosis (S1603)
         Petteri Nieminen, Finland
16.30    The role of testosterone in reproductive physiology and evolution of a mammalian
         species (S1604)
         Mikael Mökkönen, Finland


Symposium 17 (Auditorium “Donner”)

15.00 – 17.00 Orthostatic influence on cerebral blood flow - from humans to
              giraffes
              Chairs: Niels Henry Secher, Christian Aalkjaer

15.00    Brain circulation in relation to upright/supine position in the normal and injured
         brain (S1701)
         Per-Olof Grände, Sweden
15.30    Orthostatic challenges of the giraffe’s cardiovascular system (S1702)
         Emil Brøndum, Denmark
16.00    Orthostatic challenges of the snake’s cardiovascular system (S1703)
         Tobias Wang, Denmark
16.30    Debate: The upright seated position is the normal position for humans (S1704)
         Peter Norsk, Denmark
16.45    Debate: The supine position is the normal position for humans (S1705)
         Niels Secher, Denmark

                                            28
                        Saturday August 16


17.00   Transportation to the Maikkula Manor House via hotels

19.00   Congress Dinner at Maikkula Manor House




                               29
                                   Sunday August 17


Symposium 18 (Auditorium “Tigerstedt”)

08.00 – 10.00 Neurosteroids in brain function
              Chairs: Esa Korpi, Torbjörn Bäckström

08.00    Neurosteroids: neurone specific endogenous modulators of the GABA-A (S1801)
         Delia Belelli, UK
08.30    Neurosteroids in neuronal damage and repair (S1802)
         Michael Schumacher, France
09.00    Mechanisms of actions of neurosteroids in GABA-A receptor mouse model systems
         (S1803)
         Elli Leppä / Esa Korpi, Finland
09.30    Mechanisms for neurosteroid action on cognitive function from receptors to human
         disorders (S1804)
         Jessica Strömberg / Torbjörn Bäckström, Sweden


Symposium 19 (Auditorium “Granit”)

08.00 – 10.00 Testosterone and male reproductive physiology
              Chair: Jorma Toppari

08.00    Androgen signaling (S1901)
         Olli A. Jänne, Finland
08.30    How much testosterone does the man need? (S1902)
         Ilpo Huhtaniemi, UK
09.00    Developmental effects of anti-androgens on reproductive system (S1903)
         Jorma Toppari, Finland
09.30    Androgen receptor polymorphism and male reproductive health (S1904)
         Aleksander Giwercman, Sweden


10.00 – 10.30 Coffee break




                                           30
                                    Sunday August 17


Panel discussion (Saalasti Hall)

10.30 – 12.30 Physiology 2008 – future directions and threats
              Chair: Karl-Heinz Herzig

01       Bertil Fredholm – Chairman of the Noble Prize committee for Physiology and
         Medicine

02       Kristian Donner – Professor, Department of Biological and Environmental
         Sciences, University of Helsinki

03       Riitta Korpela – Vice Director of R & D, Valio. Professor, Institute of Biomedicine,
         Biomedicum Helsinki

04       Erkki Nissinen – Head of Scientific Affairs, Orion Pharma, Espoo, Finland

05       Peter Bie – President of the Scandinavian Physiological Society (SPS)

06       Ulrich Pohl – President of the Federation of European Physiological Societies
         (FEPS)

         10-minute presentations/statements by each participant followed by a podium
         discussion
         Chair: Karl-Heinz Herzig

         We hope for an active participation from the audience. Questions to the panel can
         be delivered in advance to congress desk.


12.30 – 13.00 Invitation to SPS 2009 Annual Meeting in Uppsala,
              Gunnar Flemström - President of the Meeting




13.00         Closing of the meeting




                                            31
                                            Abstracts




                                           Abstracts


Abstracts will be also published online. When citing abstracts, state abstract number (not page
number) in references. Abstract numbers are L01-L03 for state-of-the-art (plenary) lectures, S01xx–
S18xx for symposia, F01xx-F04xx, for free oral communications, and P01-P78 for posters.
Example: Acta Physiologica 2008, Vol. 193 Supplement 664: S0101.




                                                33
                                              Abstracts


                                     State-of-the-art lecture 01

L01
The action potential
STUART G
Division of Neuroscience, The John Curtin School of Medical Research, Canberra ACT 2601,
Australia

The action potential is the fundamental electrical signal used by the brain for communication
between nerve cells and for interfacing the brain with our bodies. In my lecture I will review recent
work on action potentials, including their site of generation, their modulation by subthreshold
synaptic input, and their role in synaptic plasticity.

                                     State-of-the-art lecture 02

L02
Molecular regulation of angiogenesis and lymphangiogenesis
ALITALO K
Molecular Cancer Biology Program, University of Helsinki, Finland

                                     State-of-the-art lecture 03

L03
Neurotrophic factors and their receptors in development and disease
SAARMA M
Institute of Biotechnology, University of Helsinki, Finland

In Parkinson‟s disease brain dopaminergic neurons degenerate in the midbrain. Neurotrophic factors
(NTF) promote the survival, differentiation and maintenance of neurons in developing and adult
vertebrate nervous system, and have a great potential for the treatment of neurodegenerative
diseases. The most potent NTF for dopaminergic neurons described so far are the glial cell line-
derived neurotrophic factor, GDNF and its homolog neurturin1. We have discovered a novel
conserved dopamine neurotrophic factor (CDNF) as a trophic factor for dopaminergic neurons.
CDNF and its vertebrate and invertebrate homologs the mesencephalic astrocyte-derived neuro-
trophic factor (MANF) are secreted proteins with eight conserved cysteine residues defining a novel,
evolutionarily conserved NTF family. In vivo, CDNF protected and repaired the nigrostriatal
dopaminergic neurons in the rat 6-hydroxydopamine (6-OHDA) model of Parkinson‟s disease2. Our
results suggest that CDNF might be beneficial for the treatment of Parkinson‟s disease. Little is
known about the neurotrophic factors in Drosophila. We have described a novel NTF in Drosophila,
DmMANF and demonstrated that it is essential for the of maintenance dopaminergic neurites and
dopamine levels. The rescue experiments confirm DmMANF as a functional ortholog of the human
MANF gene thus opening the window for comparative studies of this novel family of NTF.
1
  Bespalov M., Saarma M. (2007) GDNF receptor complex is an emerging drug target. Trends in
Pharmacological Sciences 28 (2), 68-74. 2Lindholm, P. et al. and Saarma, M. (2007) Nature, 448,
73-77.


                                                  35
                                              Abstracts


            Symposium 01: Physiological phenotype of genetically modified animals

S0101
Physiological monitoring of microcirculation in genetically modified animals
CURRY FE
Department of Physiology and Membrane Biology, School of Medicine, University of California
Davis, One Shields Avenue, Davis, CA 95616,USA, and Department of Biomedicine, University of
Bergen, Norway

Most detailed measurements of microvascular function in animal models of the microcirculation
involve direct observation on exposed microvascular beds to measure functional parameters
describing network geometry, local blood flow, permeability in individual microvessels, and the
interaction of inflammatory cells with the microvessel walls. Although these invasive approaches
continue to be used for the physiological phenotyping of genetically modified animals, they are often
limited to specific tissue (which may not express the phenotype) and are especially difficult to apply
when the same animal must be repeatedly studied as the phenotype develops. Based on lessons
learned from the more invasive approaches, we have begun to evaluate the use of magnetic
resonance imaging (MRI), long- wavelength optical tracers, and microPET to measure independent
changes in microvascular blood volume, increased vascular permeability, and localized
accumulation of inflammatory cells in a range of tissues in mice over periods of hours, days or
weeks. Projects begun in collaboration with laboratories at Bergen and Davis include the use of MRI
to measure blood to tissue clearances normalized for changes in local vascular volume using a
gadolinium based contrast agent with an apparent MW of 35 kDa, and the use of quantitative
fluorescent imaging of albumin labeled with the long wavelength (Alexa) dyes to distinguish
endothelial, neutrophil and platelet dependent mechanisms that regulate wound healing over periods
of days.

S0102
Physiological function of cytoskeleton and motor proteins in muscle studied in transgenic mice
and zebrafish larvae
ARNER A
Div. Genetic Physiology, Dept Physiology and Pharmacology, Karolinska Institutet v Eulers väg 8
SE 171 77 Stockholm, Sweden

Contraction of muscle is due to interaction between actin and myosin. The contractile system is
anchored to the cytoskeleton. Physiological function of myosin motors and cytoskeleton was
examined using transgenic mice lacking the intermediate filament protein desmin (Des-/- mice) and
smooth muscle myosin heavy chain (SMMHC-/- mice). Desmin is normally expressed in all muscle
types, with tissue variability. Des-/- mice are viable, but have a cardiomyopathy with increased heart
weight and decreased systolic function. Lack of desmin resulted in a lower active force generation
and altered contractile filament lattice structure and cellular compliance. We conclude that desmin
plays a role in supporting active tension possibly by affecting force transmission or sarcomere
alignment. SMMHC-/- mice survive for about 3 days after birth, and their smooth muscles can
contract due to non-muscle myosins. We found that non-muscle myosins can form thick filaments
and support slow contractions. Non-muscle myosin is also expressed in some adult smooth muscles
(e.g. aorta) and can have a physiological role in sustained contractions of vascular muscle. Due to its


                                                  36
                                              Abstracts

efficient breeding, rapid development and characterized genome the zebrafish (Danio rerio) has
become an interesting vertebrate model organism in developmental and systems biology. The larvae
stage (5-7 days), when the larva is about 3 mm long, is of particular interest since the gene
expression can be manipulated by an antisense approach. We have recently developed a system for
studies of muscle function in the larvae and report from our first physiological characterization of
mechanical and structural properties of zebrafish larvae muscles.

S0103
Structural and regulatory roles of conserved collagens
PIHLAJANIEMI T
Institute of Biomedicine, Medical Biochemistry and Molecular Biology, University of Oulu, Oulu,
Finland

The extracellular matrix (ECM) plays a crucial role in controlling cell differentiation and function in
multicellular animals. It is continually remodelled and synthesised by cells in response to
environmental factors such as physical force, hypoxia, trauma and infection. Cells receive survival
and positional information from the ECM and it is now evident that ECM biology permeates all
aspects of cellular function. Moreover, the ECM is associated with a wide spectrum of common
human diseases including arthritis, skeletal deformity, atherosclerosis, diabetes, fibrosis, cancer and
poor wound healing. Collagens play a dominant structural role in maintaining the integrity of tissues.
However, compelling evidence shows that collagens can also serve as a reservoir for growth factors
and modulate cell signaling critical for tissue morphogenesis and homeostasis. Among the 28
collagens described in vertebrates, two are known to be conserved between mammals, fish, flies and
worms, namely the basement membrane (BM) collagen IV and a XV/XVIII homologue. A collagen
XVIII fragment, endostatin, may have a role in controlling blood vessel formation in tumours as
described by Judah Folkman and coworkers. Collagen XVIII is expressed as three N-terminal variant
polypeptides, the short, the middle and the long, that arise from the use of two alternative promoters
and alternative splicing, whereas collagen XV lacks the N-terminal variants. The long form contains
a cysteine-rich frizzled (Fz) domain, which may be involved in Wnt signalling and inhibition of
tolloid proteinases. Our data on Col18a1-/- mice indicate that lack of collagen XVIII delays apoptotic
regression of hyaloid vessels. On the other hand, the retinal vessels form poorly, possibly because
the heparan sulphate side chains of this collagen are needed to provide VEGF for endothelial cells.
Our data thus suggest that collagen XVIII participates both in the inhibition and stimulation of vessel
growth. Our recent data also indicates that endostatin can affect formation of lymphatic vessels and
metastasis in a squamous cell carcinoma model. Moreover, endostatin fragments appear to
counteract in a dominant negative manner the physiological functions of the endostatin domains of
the full-length collagen XVIII. Altogether, the distinct roles in physiological and pathological
processes of the complex ECM molecule, collagen XVIII, will be discussed.

S0104
Mouse models for skin development and skin diseases
BRAKEBUSCH C
BRIC, University of Copenhagen, 2200 Copenhagen, Denmark

Genetically modified mice are excellent models to study the function of specific genes in skin
development and in skin diseases. Using mice with targeted mutations of Rho GTPase genes as

                                                  37
                                               Abstracts

example we will demonstrate analysis of skin development and maintenance, wound healing, skin
inflammation and tumor formation and show, how mechanistical information can be extracted from
these in vivo models. Rho GTPases are major regulators of the actin cytoskeleton, but more recently
were found to control also other cellular processes such as proliferation, apoptosis, cell-cell contact,
cell polarity and endocytosis. Our investigations with mutant mice reveal important roles for Rho
GTPases in the fate decision of hair follicle progenitor cells, in the maintenance of cell-cell contacts,
reepithelialization and inflammatory and neoplastic skin diseases.


S0105
Use of imaging techniques in the analysis of mouse phenotypes
THORSEN F
University of Bergen, Department of Biomedicine, Jonas Lies vei 91, 5009 Bergen, Norway

The mouse has become one of the most popular mammals for biomedical research. With the growing
amount of experimental and genetic mouse models of human disease, there is a need for efficient
and reliable methods for characterizing abnormalities in mouse anatomy and physiology. Common
tumour imaging modalities for use with small animals include magnetic resonance imaging (MRI),
computed tomography (CT), positron emission tomography (PET), single photon emission
computed tomography (SPECT), bioluminescent imaging and ultrasound. Each of these modalities
has its advantages and limitations, which will be briefly outlined here. The use of MRI provides a
promising technology providing informative and meaningful measures in a variety of mouse models,
and several current MR imaging techniques of small animals will be presented. Differences in
contrast in soft tissues depend on endogenous differences in water content, relaxation times and
diffusion characteristics of the tissue. The specificity of MRI can be further increased by exogenous
contrast agents, such as gadolinium chelates, which have been successfully used for imaging of
hemodynamic parameters including blood perfusion and vascular permeability. Cellular MR
imaging for visualising targeted cells in living organisms are also performed, using cell labelling
with superparamagnetic iron oxide particles. However, due to the low sensitivity of MRI, compared
with nuclear imaging, high local concentrations at the target site are required to generate detectable
MR contrast.




                                                   38
                                              Abstracts


                         Symposium 02: Physical activity and health -
                      Human physiology in changing thermal environments

S0201
Climate indices for human performance and health in outside conditions
PARSONS K
Human Thermal Environments Laboratory, Department of Human Sciences, Loughborough
University, Loughborough, Leicestershire LE11 3TU, UK

The physiological and behavioural responses of people to outside thermal conditions will be
influenced by air temperature, radiant temperature including the sun, air velocity (wind) and
humidity (also related to fog, rain, frost, snow etc). The responses will also depend upon the clothing
worn and the activity carried out. A climate index is a single number that integrates the effects of the
above factors to represent the thermal strain caused by the climate. The aim of the research presented
in this paper was to evaluate existing thermal indices for use in outdoor climates. Laboratory
experiments into the effects of solar radiation and field studies measuring weather conditions as well
as physiological and subjective responses of people are presented. Comparison of the responses of
people with values of thermal indices (Wet Bulb Globe Temperature (WBGT); Predicted Mean Vote
(PMV); Wind Chill Index (WCI); provided an indication of the validity of current thermal indices.
The results of a series of laboratory studies showed that the PMV index calculated using mean
radiation temperature in the shade, and corrected for solar radiation (increase by 1 PMV scale unit
for every 200Wm-2 of direct solar radiation) provides an index useful in the sun (PMVsolar). The
field studies demonstrated that both the PMVsolar and WBGT indices provide good correlation with
human response to outside conditions.

S0202
Heat, hydration and exercise
MAUGHAN RJ, WATSON P, SHIRREFFS SM
School of Sport & Exercise Sciences, Loughborough University LE11 3TU, UK

It is now common for major athletic contests to take place in conditions of high heat and humidity.
This poses a major challenge for competitors, as the performance of both physical and mental tasks
can be adversely affected by heat and by dehydration. The increased sensation of effort during
exercise in the heat is also a challenge to the promotion of health-related physical activity in hot
climates. There are well-recognised effects of heat and hydration status on the cardiovascular and
endocrine systems, and these effects play a role in the decreased performance experienced in the
heat. However, growing evidence from various experimental models suggests that the primary
effects of high ambient temperature and dehydration on performance may be mediated, at least in
part, by effects on the central nervous system. This seems to be a consequence of changes in
serotonergic and dopaminergic function, with pharmacological manipulation of these
neurotransmitters demonstrated to influence exercise performance. Recent evidence suggests that the
integrity of the blood brain barrier may be compromised by combined heat stress and dehydration,
and this may also play a role in limiting performance in the heat. Coping strategies include increased
aerobic fitness, prior exposure to exercise/heat stress resulting in acclimatisation, pre-exercise
cooling, and hydration. Provision of fluids of an appropriate composition can prevent dehydration
and can greatly reduce the adverse effects of heat stress.


                                                  39
                                              Abstracts



S0203
Cardiovascular regulation in cold
MÄNTYSAARI M
Aeromedical Centre, Centre for Military Medicine, P.O. Box 50, 00301 Helsinki, Finland

This presentation reviews the function of cardiovascular regulation in cold environment. The main
hemodynamic effects of cold exposure are skin vasoconstriction, elevation in blood pressure and
centralisation of circulating blood volume. The hemodynamic effects of acute and chronic cold
exposure are discussed as well as the role of habituation in repeated cold exposures. Also the
differences between skin and whole body cooling are considered. The roles of central and local
circulatory regulation mechanisms in responses to cold exposure are reviewed, including autonomic
nervous system, renin-angiotensin-system, nitric oxide, endothelin system, arginine vasopressin,
corticotrophin releasing factor, and adrenocorticotrophin hormone. The interplay between the
different regulatory mechanisms is also discussed.

S0204
Neuromuscular performance in cold
OKSA J
Finnish Institute of Occupational Health, Oulu, Finland

Exposure to cold ambient temperature may induce subnormal body and muscle temperatures leading
to decreased neuromuscular performance. In general, decrease in dynamic performance after cooling
is in the order of 2 - 10 % × °C-1 decrease in muscle temperature. However, even bigger values have
been reported, during drop jump exercise the highest decrease in performance was 17 % × °C-1. This
implies that exercise, which is very fast and efficiently utilises elasticity of the working muscles is
very susceptible for cooling. A dose–response relationship between muscle temperature and
performance can be found. Whether the muscle is passively cooled or actively rewarmed after
cooling is of no importance; the predominant factor in determining the outcome of muscular
performance is muscle temperature. In addition to decreased muscular performance cooling has also
a profound effect on functional properties of skeletal muscle. The rate of tension development and
relaxation, as well as the velocity of muscle contraction itself, shortening and lengthening, is also
slower in a given time when muscle tissue is cooled resulting in a less powerful contraction of a
muscle. Force production is regulated peripherally and/or centrally. There is evidence that the
suppressed T-reflex amplitude is caused by decreased activity of the muscle spindles and thus
decreased gamma motoneuron excitability and these changes may lead to decreased force
production. On the other hand, it also has been shown that during low-intensity repetitive work in
cold, stretch reflex responses are enhanced in relation to thermoneutral responses. This suggests that
the increased strain of the working muscles were met by increasing reflex activity. Therefore, more
muscle fibres are recruited to maintain the given work level in the cold.




                                                  40
                                              Abstracts


Symposium 03: Neuronal excitability: synaptic vs. extrasynaptic influence (MTP-symposium)

S0301
Voltage-sensor modulation in K channels regulates neuronal excitability
ELINDER F
Department of Clinical and Experimental Medicine, Linköping University, Sweden

The size of the movement and the molecular identity of the moving parts of the voltage sensor of a
voltage-gated ion channel are debated. In the helical-screw model, the positively charged fourth
transmembrane segment S4 slides and rotates along negative counter charges in S2 and S3, while in
the paddle model, S4 carries the extracellular part of S3 (S3b) as a cargo. By introducing pairs of
cysteines in S3b and S4 we show that S4 slides 16-26 Å along S3b. These data are not compatible
with the voltage-sensor-paddle model, but support the helical-screw model. Knowing the voltage
sensor mechanism we can analyze substances affecting excitability at a molecular level.
Polyunsaturated fatty acids (PUFAs) have beneficial effects on epileptic seizures and cardiac
arrhythmia. We have found that ω-3 and ω-6 all-cis-PUFAs affect the voltage dependence of a K
channel by attracting the voltage sensor S4 to an extracellular position, thereby opening the channel.
Our data suggests that fatty acid tails with two or more cis double bonds are required to place the
negative carboxylate charge of the PUFA in a position to affect the channel‟s voltage dependence.
We propose that charged lipophilic compounds could play a role in regulating neuronal excitability
by electrostatically affecting the channel‟s voltage sensor. This provides a new approach for
pharmacological treatment which is voltage sensor pharmacology

S0302
Functional role of tonic GABA in the hippocampus
WALKER MC
Institute of Neurology, University College London, London, UK

GABA(A) receptor mediated signalling can be divided into fast synaptic (phasic) transmission and
tonic GABA(A) receptor signalling in which extrasynaptic receptors are activated by ambient,
extracellular GABA. Since extracellular GABA concentrations can change in both health and
disease, then tonic GABA currents are variable, responding and contributing to different brain states.
The functional role of tonic currents is still unclear. We have recently found that tonic GABA(A)
receptor currents in pyramidal cells are outwardly rectifying. This has an important effect on the
function of tonic inhibition. In contrast to studies in the cerebellum, hippocampal tonic currents alter
the offset rather than the gain of pyramidal cell input-output functions. The magnitude of tonic
inhibition depends not only upon the concentration of extracellular GABA but also on the expression
of extrasynaptic receptors. We have demonstrated that the receptors mediating tonic currents show
adaptive plasticity in epilepsy. This may compensate for increased network excitability at the
expense of cognitive function. Thus tonic GABA(A) receptor mediated currents in the hippocampus
show adaptive plasticity and play an integral role in regulating neuronal and network excitability.




                                                  41
                                             Abstracts

S0303
P2X mediated transmission in neuronal-glial networks
VERKHRATSKY A
Faculty of Life Sciences, The University of Manchester, Manchester M13 9PT, UK and Institute of
Experimental Medicine, ASCR, Videnska 1083, 142 20 Prague 4, Czech Republic

The ATP, discovered in 1929 by Karl Lohman, Cyrus Hartwell Fiske and Yellagaprada SubbaRow,
acts as an important extracellular signalling molecule. Purinoreceptors, represented by ionotropic
P2X and metabotropic P2Y receptors are arguably the most abundant receptors in living tissues and
are expressed in neural cells in both peripheral and central nervous system. In the CNS, ATP can be
released from synaptic terminals, either on its own or together with other neurotransmitters.
Furthermore, ATP also acts as an important mediator in neuronal-glial communications, as glial cells
are endowed with numerous ATP receptors, which trigger Ca2+ signalling events and membrane
currents in both macro- and microglia. In addition ATP can be released from astroglial cells thereby
acting as a mediator of glial-glial and glial-neuronal signalling.

S0304
Tonically pre-bound extrasynaptic NMDA receptors
GREBENYUK S, SEMYANOV A
Unit of Extrasynaptic Transmission, RIKEN Brain Science Institute (BSI), Wako-shi, Japan, and
Department of Neurodynamics and Neurobiology, UNN/IAP RAS, Nizhny Novgorod, Russia

High affinity extrasynaptic NMDA and GABAA receptors both can detect ambient concentrations of
endogenous agonists. GABAA receptor mediated tonic current is present at physiological conditions
while tonic current mediated by NMDA receptors could be obtained only when receptors are relived
from voltage-dependent channel block by Mg2+. Using two-photon calcium imaging in slices we
demonstrated that extrasynaptic NMDA receptors in dendrites of hippocampal pyramidal neurons
are pre-bound to ambient glutamate and mediate a proportion of calcium transients during
backpropagating action potentials. This happens because of transient relief of Mg2+ block at the time
of action potential. Local glutamate uncaging paired with backpropagating action potential further
increased NMDA receptor dependent part of calcium transient. Such „extrasynaptic coincidence
detection‟ could be important for synchronisation of simultaneously firing neurons. In other words
glutamate spillover from neighbouring synapses or glutamate release by astrocytes can be sensed in
larger extent by those neurons which fire at the same time.

S0305
Chloride concentration changes determine current time course and mediate interaction
between GABA receptors and glycine receptors
JOHANSSON S
Department of Integrative Medical Biology, Section for Physiology, Umeå University, SE-901 87
Umeå, Sweden

Desensitization of ligand-gated ion channels plays a critical role for neuronal signalling. Common
GABAA receptors and glycine receptors show significant desensitization with a time course that
depends on receptor subunits and on ligand concentration. The time course of recorded current that
is usually taken to represent desensitization may, however, be critically affected by changes in

                                                 42
                                               Abstracts

intracellular ion concentrations. Here, I describe an analysis designed to separately estimate the time
course of conductance and the changes in intracellular Cl– concentration, [Cl–] i, during activation of
native GABAA- and glycine-receptors in preoptic neurons from rat. In contrast to the prevailing
view, it is shown that changes in [Cl–]i are critical for the decay of current in the presence of either
GABA or glycine while changes in conductance play a minor role only. Conductances decay with
time constants of several seconds and in some cells do not decay below the value at peak current,
during 20-s agonist application. In contrast to recent reports, it is also shown that apparent cross-
desensitization of currents evoked by GABA and by glycine is caused by changes in [Cl–]i. By
taking the cytosolic volume into account and numerically computing membrane currents and
expected changes in [Cl–] i, a theoretical framework is provided for the observed effects. Modelling
diffusional exchange of Cl– between cytosol and patch pipettes shows that considerable changes in
[Cl–] i may be expected and cause rapidly decaying current components in conventional whole-cell
or outside-out patch recordings. The findings imply that a re-evaluation of the desensitization
properties of GABAA- and glycine-receptors is needed.




                                                   43
                                             Abstracts


                   Symposium 04: Neurotransmission and neuromodulation I

S0401
GABA-A receptors mediating tonic inhibition in the neocortex
JENSEN K
Synaptic Physiology Laboratory, Institute of Physiology and Biophysics 1160, University of Aarhus,
Denmark

GABA (gamma-aminobutyric acid) is the major inhibitory neurotransmitter in the mammalian
central nervous system. GABA activates GABA-A receptors, which are pentameric chloride
channels localized at synapses, and at locations away from the synaptic cleft so called extrasynaptic
receptors. At synapses, GABA-A receptor activation leads to fast phasic signaling on the millisecond
timescale shaped by the rapid GABA transient. Extrasynaptically, elevated extracellular GABA
levels may activate receptors on a slower timescale leading to tonic inhibition. Prototypical
extrasynaptic GABA-A receptors contain combinations of alpha4 and delta subunits, which confer a
high affinity for GABA and little desensitization. We investigated tonic inhibition in mouse
neocortex using whole-cell patch-clamp recordings in brain slices. In layer 2/3 pyramidal cells,
elevated GABA levels led to significant a tonic current revealed by the GABA- A receptor
antagonist SR95531. THDOC, a delta- subunit selective GABA-A modulator and neurosteroid,
strongly enhanced the tonic current, while the delta-subunit selective agonist THIP also induced a
clear tonic current. The benzodiazepine zolpidem had no effect on tonic GABA-A mediated
currents. In layer 5 of the neocortex, which has a lower level of delta- subunit expression, pyramidal
cells displayed a diminished tonic current. Finally, regular- spiking somatostatin-positive
interneurons were completely devoid of tonic inhibition in response to GABA, THIP, or THDOC. In
conclusion, delta-subunits contributes to the tonic inhibition in mouse neocortex in a cell- type
specific manner. It is likely that tonic GABA-A receptor mediated inhibition and its enhancement by
neurosteroids will only affect certain neurons in the cortical network.

S0402
GABA mediated inhibition in rat hippocampus is modulated by insulin
BIRNIR B, JIN Z
Lund University, CRC, Entry 72, house 91, level 11 UMAS, 205 02 Malmö, SWEDEN

Hippocampal neurons can sense and respond to changes in metabolic signals such as insulin and
glucose. GABA (gamma-aminobutyric acid) and its receptors have been shown to have a role in the
central control of metabolic homeostasis. The aim of our study was to investigate effects of insulin
on GABA-A receptors-mediated inhibition in rat hippocampal CA1 pyramidal neurons. Rat
hippocampal slices (P16-P22) were prepared and incubated with low concentration of insulin for 1,
2, and 3 hours. Standard whole-cell recordings and quantitative PCR were performed. Insulin
incubation regulated both phasic and tonic currents mediated by GABA-A receptors in a temporal
manner. PCR results from the CA1 region showed altered mRNA expression of alpha4, alpha5 but
not delta GABA-A receptor subunits. Our results suggest that low concentration of insulin
potentiates GABA-generated inhibition by modulating both synaptic and extrasynaptic GABA
currents.




                                                 44
                                             Abstracts

S0403
Glutamatergic adaptation of dopamine neurons to single doses of benzodiazepines
HEIKKINEN AE, KORPI ER
University of Helsinki, Institute of biomedicine / pharmacology

Initial effects of drugs of abuse seem to converge on the mesolimbic dopamine pathway originating
from the ventral tegmental area (VTA). Many drugs of abuse, already after a single dose, are able to
modulate the glutamatergic transmission activating the VTA dopamine neurons, which could
represent a critical early step in the development of addiction. Ligands acting on the benzodiazepine
site of the inhibitory gamma-aminobutyric acid type A (GABA-A) receptors are known to be
rewarding in animal models and have abuse liability in humans, but notably little evidence exists that
the mesolimbic dopamine system would be involved in their effects. We have now found that,
similarly to classical drugs of abuse like morphine and ethanol, single in vivo doses of
benzodiazepine-site agonists induce a modulation in the glutamatergic transmission of VTA
dopamine neurons. This was seen 24 h after the injection as an increase in the ratio between AMPA
and NMDA receptor-mediated excitatory currents using whole-cell patch-clamp configuration in
mouse VTA slices. The effect was due to increased frequency of spontaneous miniature AMPA
receptor-mediated currents. It lasted at least 3 days after the injection of diazepam, and it was
prevented by co-administration of the benzodiazepine-site antagonist flumazenil or the NMDA
receptor antagonist dizocilpine. Also a single injection of the GABA-A receptor alpha-1 subunit-
preferring benzodiazepine-site ligand zolpidem produced an increase in the AMPA/NMDA ratio in
VTA dopamine neurons. These findings suggest a role for the mesolimbic dopamine system in the
initial actions of and on neuronal adaptation to benzodiazepines.




                                                 45
                                             Abstracts


        Symposium 05: Modern imaging technologies in the study of human physiology

S0501
Use of Doppler ultrasound and coronary CT to study coronary circulation and vascular
endothelial function in man
HARTIALA J
Clinical Physiology and Nuclear Medicine, Turku University Hospital, 20500 Turku, Finland

Recent development in ultrasound technology has made it possible to visualise noninvasively human
coronary arteries (including diameter changes) by using transthoracic echocardiography . Moreover,
Doppler technology allows measurement of flow velocities in the coronary arteries. This makes it
possible to study human coronary reactivity in various provocations. This has earlier been possible
only in peripheral arteries like in studies of vascular endothelial function by studying flow mediated
dilatation of brachial artery. Computed X-ray tomography of the coronary arteries has been widely
taken in clinical use recently. The method yields anatomic images of coronary arteries comparable to
invasive coronary angiography, but does not allow evaluation of the pathophysiological
consequences of possible flow obstruction due to coronary calcifications. Therefore, it often leads to
nuclear studies of myocardial perfusion.

S0502
Evolving magnetic resonance imaging (MRI) and spectroscopy (MRS) techniques in the study
of human physiology
BORRA R
Medical Imaging Centre of Southwest Finland & Turku PET Centre, PO Box 52, FIN-20521 Turku,
Finland

Magnetic Resonance Imaging (MRI), providing high resolution anatomical images, is the most
widely know application of magnetic resonance scanners. However, magnetic resonance also offers
great opportunities for characterization of tissue metabolism, using Magnetic Resonance
Spectroscopy (MRS). In the past decade great advances in MRS technology have been made,
especially in the development of Proton Magnetic Resonance Spectroscopy (1H MRS) protocols,
some of which already have become part of clinical practise. Objective: In this lecture an overview
is given of current possibilities for non-invasive fat content quantification of the myocardium,
skeletal muscle and liver by 1H MRS. In addition, the development en current status of
hyperpolarized contrast agents and also of combined Magnetic Resonance (MR) and Positron
Emission Tomography (PET) imaging (MR-PET) will be discussed. These (near) future techniques
potentially offer a wide range of new MRI and MRS applications. Conclusions: Several Magnetic
Resonance based methods are currently available for the study of tissue metabolism. The rapid
ongoing development of future MR applications has the potential to open a whole new field of non-
invasive and real-time metabolic imaging of human physiology.

S0503
Molecular imaging of metabolism using positron emission tomography
NUUTILA P
Turku PET Centre, University of Turku, Finland



                                                 46
                                              Abstracts

Positron emission tomography (PET) represents an advanced non-invasive scintigraphic imaging
technology and provides a possibility for quantitative characterisation of physiologic processes in
human tissues. PET imaging is based on the use of short-lived positron emitting radioisotopes such
as carbon-11, oxygen-15 and fluorine-18 and the detection of two photons created in an annihilation
reaction between a positron and a tissue electron. PET combined with tracer kinetic models
measures blood flow, membrane transport, metabolism, ligand interaction and recently also gene
expression non-invasively and quantitatively. Numerous studies on muscle metabolism and
perfusion in healthy subjects and in patients with metabolic disorders are conducted to elucidate their
regulation and interactions. PET combined with [18F]-2-fluoro-2-deoxy-D-glucose ([18F]FDG) has
been widely employed to measure glucose uptake and phosphorylation and muscle and other key
organs. In addition, fatty acid uptake and oxidation has been studied. The hybrid PET/CT scanners
enable correlation of anatomic and functional information. The sequential scanning procedures
including cardiac, hepatic, adipose tissue and cerebral scans are able to give an integrative view of
human physiology. Examples of some recent studies on metabolic syndrome and obesity and effects
of weight loss are reviewed. Because other methods (e.g. acute exercise and drug interventions,
glucose-insulin clamp technique, stable tracer infusions, MRI/MRS) can be combined with PET, it
serves an unique tool the assessment of physiology in experimental and clinical settings.

S0504
Abstract missing




                                                  47
                                                Abstracts


     Symposium 06: Gastrointestinal barrier and transport functions in health and disease

S0601
Abstract missing

S0602
Intragastric nitric oxide and gastric mucosal defense
PETERSSON J
Medical Cell Biology, Uppsala University, Uppsala, Sweden

The human stomach normally contains high levels of bioactive nitric oxide. This NO derives from
salivary nitrate that is converted to nitrite by oral bacteria and thereafter non-enzymatically reduced
in the acidic gastric lumen to NO. Nitrate is a common component in vegetables, and after ingestion
it is absorbed in the small intestine. Interestingly, circulating nitrate is then concentrated by the
salivary glands. Hence, intake of nitrate-rich vegetables results in high levels of NO in the stomach.
The physiological effect of the high concentration of NO gas normally present in the gastric lumen
has been hitherto unknown and has therefore been the focus of my research. We have shown that NO
produced in the gastric lumen after nitrate ingestion increases gastric mucosal blood flow and the
thickness of the firmly adherent mucus layer in the stomach. The blood flow and mucus layer are
essential defense mechanisms that protect the mucosa from luminal acid and noxious agents. We
have demonstrated that a nitrate-rich diet protects against Nonsteroidal anti-inflammatory drugs-
induced gastric damage, as a result of the increased formation of NO in the stomach. We have also
shown that the gastroprotective effect attributed to nitrate depends completely on conversion of
nitrate to nitrite by the bacterial flora, colonizing the tongue, and that the oral microflora therefore is
important in regulating physiological conditions in the stomach. This research challenges the current
dogma that nitrate intake is hazardous, and on the contrary suggests that dietary nitrate plays a direct
role in regulating gastric homeostasis. It is likely that a sufficient supply of nitrate in the diet
together with the oral microflora is important for preventing pathological conditions in the
gastrointestinal tract.

S0603
Congenital chloride diarrhea - a Finnish disease serves as a paradigm for electroneutral NaCl
absorption
LAMPRECHT G, GACO V, SCHÄFER J, TURNER JR, GREGOR M
University of Tübingen, Dept. of Medicine - Gastroenterology, Otfried-Müller-Str. 10, 72076
Tübingen, Germany

Introduction: Congenital chloride diarrhea is a rare disease endemic in Finland. It is due to loss of
function of the anion exchanger DRA (down regulated in adenoma, SLC26A3), which is involved in
electroneutral NaCl absorption in the ileum and colon. Inhibition of NaCl absorption occurs in other
diarrheal diseases and may involve elevated [Ca]i. It is unknown whether DRA is inhibited by [Ca]i
or whether its function follows the inhibition of NHE3, to which it is functionally and structurally
coupled. Aim: Is DRA inhibited by elevated [Ca]i? Does this involve changes in cell surface
expression? What is the role of the C-terminal PDZ interaction motif ETKF? Methods: In EGFP-
DRA or EGFP-DRA-ETKFminus transfected HEK and Caco-2/BBE cells [Ca]i was elevated by
4Br-A23187 or by 100 µM UTP. Surface expression was quantified by biotinylation. Results: In


                                                    48
                                              Abstracts

DRA-transfected HEK and Caco-2/BBE cells DRA is inhibited by elevated [Ca]i (100 sec 4Br-
A23187) independently of its PDZ interaction (45 %, P< 0.05). With a time delay surface expression
is reduced (0, 10, 30 min: 34 %, 22 %, 10 %, p< 0.05). In DRA but not in DRA-ETKFminus
transfected Caco-2/BBE cells UTP inhibits DRA (15 %, p < 0.05). HEK cells have a lower
expression of the PDZ proteins E3KARP and PDZK1 than Caco-2 cells. Transient transfection of
HEK/EGFP-DRA cells with PDZK1 establishes inhibition by UTP. Conclusion: At non-physiologic
elevations of [Ca]i by 4Br-A23187 DRA is inhibited and its surface expression is reduced
independently of its PDZ interaction. Physiologic elevation of [Ca]i by UTP causes PDZ dependent
inhibition of DRA in Caco-2/BBE cells. This can be recapitulated in HEK cells that are transfected
with PDZK1 suggesting that this adapter protein plays an important role in the transduction of the
Ca signal.

S0604
Regulatory aspects of the paracellular barrier: special role of claudin-2 and -8
AMASHEH S, SCHULZKE JD, FROMM M
Charité Campus Benjamin Franklin, Institute of Clinical Physiology, Hindenburgdamm 30, 12200
Berlin, Germany

Objective: Recent molecular identification of tight junction constituents has paved the way for
analyses of structure, function and regulation of the barrier. Whereas perturbation and internalisation
of tight junction proteins away from tight junction strands have been reported, so far information
concerning induction of tight junction proteins are relatively scarce. In our studies, we have analysed
effects of induction of sodium transport and of inflammatory processes on tight junction regulation,
in vitro. Methods: Preparations of human colon (inflamed or non-inflamed) and monolayers of the
epithelial colon cell line HT-29/B6 grown on permeable supports were mounted in Ussing chambers
and corticosteroids and proinflammatory cytokines were added, respectively. After incubation,
molecular analyses including confocal laser-scanning microscopy, real time PCR and
immunoblotting were performed on selfsame cells and tissues. Results: After induction and
subsequent inhibition of the epithelial sodium channel employing corticosteroids and the specific
blocker amiloride, an induction of occludin and claudin-8 was observed. Claudin-8 was induced on
transcriptional level, and Na+-entry into the cell was a prerequisite for this effect in both, colon
preparations and epithelial cells. In a separate approach, tight junction proteins were monitored in
inflamed colon epithelia of patients with Crohn's disease and ulcerative colitis. These tissues show
an increased expression of claudin-2, which could be evaluated by incubation of the epithelial colon
cell line with proinflammatory cytokines. Conclusions: In these studies, novel mechanisms of barrier
regulation are reported: a synergistic effect of transport and barrier function during physiological
Na+ absorption, and the perturbation of barrier properties in IBD, which can be attributed to direct
effects of proinflammatory cytokines on claudin-2.

S0605
The role of the NHERF family of PDZ-scaffolding proteins in the regulation of salt and water
transport: lessons learned from knockout mice
SEIDLER U
Department of Gastroenterology, Hannover Medical School, Hannover, Germany




                                                  49
                                            Abstracts



The four members of the NHERF family of PDZ-adapter proteins bind to a variety of membrane
transporters and receptors, and modulate membrane expression, mobility, interaction with other
proteins, and the formation of signalling complexes. All four family members are expressed in the
intestine. The CFTR anion channel and the Na+/H+ exchanger NHE3 are two prominent bindings
partners to this PDZ-adapter family, which are also known to be key players in the regulation of
intestinal electrolyte and fluid transport. Experiments in heterologous expression systems have
provided a number of mechanistic models how NHERF proteins can interact with some of their
targets on a molecular level. Recently, however, NHERF1,2 and 3 knockout mice have become
available, and this presentation will highlight the findings on electrolyte and fluid transport
regulation in the native intestine of these mice.




                                                50
                                              Abstracts


                   Symposium 07: Neurotransmission and neuromodulation II

S0701
Monocarboxylate transporters at brain synapses
BERGERSEN LH
Department of Anatomy and CMBN, University Of Oslo, Norway

Intercellular monocarboxylate transport is important, particularly in tissues with high energy
demands, such as brain and muscle. In skeletal muscle, it is well established that glycolytic fast
twitch muscle fibres produce lactate, which is transported out of the cell through the
monocarboxylate transporter MCT4. Lactate is then taken up and oxidized by the oxidative slow
twitch muscle fibres, which express MCT1. In the brain it is still questioned whether lactate
produced in astrocytes is taken up and oxidized by neurons upon activation. Our studies have
reported that astrocytes express MCT4, whereas neurons express MCT2. By comparing the
localizations of MCTs in oxidative and glycolytic compartments I here give support to the idea that
there is a lactate shuttle in the brain similar to that in muscle. This conclusion is based on studies
using high resolution immunocytochemical methods at the light and electron microscopical levels.

S0702
Glutamate release from astrocytes
GUNDERSEN V
Department of Anatomy and the CMBN, POB 1105 Blindern, 0317 Oslo, Norway

Glutamate is the established transmitter at most brain synapses. Recent evidence has shown that
glutamate can be released from astrocytes. Here we combine electron microscopic
immunocytochemistry, live imaging techniques and patch clamp electrophysiology to show that
astrocytes contain a vesicular compartment competent of glutamate release and that this astrocyte
derived glutamate can increase the activity of adjacent excitatory synapses by activation of
presynaptic NMDA 2B receptors. Activation of astrocytic P2Y1 receptors elicits astrocytic
glutamate release and the astrocyte dependant changes in synaptic activity during neuronal activity.

S0703
Abstract missing

S0704
Abstract missing

S0705
Developmental aspects of short- and long-term plasticity in the hippocampus
HANSE E, ABRAHAMSSON T, ANDERSSON M, RIEBE I
Dept of Physiology, University of Gothenburg, Box 432, 405 30 Goteborg, Sweden

Glutamate synapses in the rodent hippocampus are generated at high rate between birth and puberty.
During this period there are also various adaptive changes in synaptic function, referred to as
synaptic maturation. This presentation focuses on three aspects of synaptic maturation; the
emergence of astrocyte- mediated short-term synaptic depression, the disappearance of AMPA


                                                  51
                                             Abstracts

silencing (generation of AMPA silent synapses by test pulse activation) and a change in the
properties of long-term potentiation (LTP). In response to synaptic activity astrocytes mediates a
depression that affects both the recently (within seconds) active synapses (homosynaptic depression)
and inactive, neighboring synapses (heterosynaptic depression). Both these depressions emerge
between postnatal days 10 to 20. During about the same time period AMPA silencing vanish and
long-term potentiation changes from being a de- depression, an AMPA unsilencing of AMPA
silenced synapses, to a “genuine” potentiation (i.e. not de-depression), not explained by AMPA
unsilencing. Glutamate synapses onto interneurons also express astrocyte-mediated depression.
However, in contrast to the glutamate synapses onto principal neurons, they express AMPA
silencing into adulthood, exemplifying divergent synaptic maturation.




                                                 52
                                             Abstracts


                         Symposium 08: Physical activity and health -
                       Cardiovascular diseases, diabetes, and osteoporosis

S0801
Individual differences in responsiveness to regular physical activity
RANKINEN T
Human Genomics Laboratory, Pennington Biomedical Research Center, 6400 Perkins Road, Baton
Rouge, LA 70808, USA

Regular physical activity and exercise training induce several beneficial changes in physiological
and biological pathways that lead to better physical performance and improved health outcomes.
However, it is evident that there are marked inter-individual differences in physiological changes
brought about by regular physical activity. The concept of heterogeneity in responsiveness to
standardized exercise programs was first introduced over 25 years ago. The most extensive data on
the individual differences in trainability come from the HERITAGE Family Study, where 742
healthy but sedentary subjects followed a highly standardized, well-controlled, laboratory-based
endurance-training program for 20 weeks. For example, the average increase in maximal oxygen
consumption was 384 (SD 202) mL, but the training responses varied from no change to increases of
more than 1 000 mL O2 per minute. A similar picture emerged for training-induced changes in
several other phenotypes, such as blood pressure, heart rate, stroke volume, plasma insulin and lipid
levels, and skeletal muscle traits. This high degree of heterogeneity in responsiveness to a fully
standardized exercise program was not accounted for by age, gender, or ethnic differences.
However, the high and low responses to regular exercise exhibited significant familial aggregation,
i.e., there were families with mainly low responders and others in which all family members show
significant improvements. The maximal heritability estimates for training response phenotypes have
ranged from 20 % to 60 %. These observations support the notion that individual variability is a
normal biological phenomenon, which may largely reflect genetic diversity. Understanding the
mechanisms that contribute to the variability in responsiveness to exercise training will ultimately
allow us to use physical activity more efficiently in primary and secondary prevention of chronic
diseases.

S0802
Impact of intense exercise on BMD, fall risk and 10y-CHD-risk in elderly females. Results of
the Senior Fitness and Prevention Study (SEFIP)
KEMMLER W, VON STENGEL S, ENGELKE K, BEBENECK M, KALENDER WA
Institute of Medical Physics, University Erlangen-Nürnberg, Germany

Purpose: To determine the effect of a combined exercise program that focuses on both, fracture and
CHD-risk. Methods: The Senior-Fitness and Prevention-Study (SEFIP) is a 18-month randomized
controlled exercise trial with 246 females of 65 years and older. The exercise group performed a
vigorous endurance, strength and balance training two times/week while a low intensity/volume
“wellness training” once per week for 4 x 10 weeks during the 18 months was dedicated to the
control group. Both groups were supplemented with calcium and vitamin-D. Bone Mineral Density
(BMD) was measured by Dual-Energy-X-Ray-Absorptiometry (DXA) and Quantitative-Computer-
Tomography (QCT), fall frequency and injuries were determined using standardized diaries and 10y-
CHD-risk was analyzed by risk calculator. Results: After 18 months 114 women completed the


                                                 53
                                              Abstracts

exercise group (EG), 118 subjects remained in the wellness control group (CG). BMD at the lumbar
spine (LS) and femoral neck (FN) as assessed by DXA differed significantly (p<.001) between both
groups (LS: EG: 1.8 %, p<.001 vs. WG: 0.3 %, n.s.; FN: 1.0 %, p=.055. vs. -1.1 %, p=.009). Overall
fall frequency, as well as the number of injurious falls was significantly lower in the exercise
compared with the control group (RR falls: 0.61, p=.001; RR injurious falls: 0.69, p=.02). 10y-CHD-
risk according to Wilson was significantly reduced (p=.008) in the EG (-25 %) and slightly
decreased in the CG (-4 %, n.s.). Conclusion: Our results clearly demonstrate that dedicated exercise
programs impact both, fracture and CHD-risk of the elderly people. Thus the implemen-tation of
corresponding programs may be helpful to decrease health costs of the elderly.

S0803
Obesity: Fat oxidation and insulin resistance
VENABLES MC
Exercise Metabolism Research Group, School of Sport and Exercise Sciences, The University of
Birmingham, Edgbaston, Birmingham, B15 2TT, United Kingdom

There is growing concern over the escalation in obesity, and obesity related diseases such as type 2
diabetes mellitus and cardiovascular disease within the western world. The evidence thus far
suggests that a combination of excessive fat intake and a reduced mitochondrial oxidative capacity,
brought about through a sedentary lifestyle, leads towards lipid storage. This lipid is stored not only
in adipose tissue but also within the skeletal muscle and the liver resulting in insulin resistance.
Interventions that increase fat oxidation can therefore be of clinical importance when targeting such
diseases. A large body of evidence exists indicating that exercise with or without weight loss can
lead to improvements in both fat oxidation and insulin sensitivity with the potential to reduce the
onset of type 2 diabetes mellitus. However weight loss without exercise, although it can confer
improvements in insulin sensitivity, does not convey any adaptations that allow for an increase in fat
oxidation. Potential mechanisms through which it is thought exercise enhances glucose tolerance and
insulin sensitivity are thought to include; an increase in oxidative capacity and fat oxidation, an
increase in intra muscular triglyceride accumulation, and a decrease in fatty acid intermediates such
as long chain acyl-CoA‟s, diacylglycerol and ceramides. More recent evidence has suggested a role
for the involvement of novel isoforms of protein kinase C (nPKC) in obesity associated insulin
resistance.

S0804
Abstract missing




                                                  54
                                              Abstracts


                     Symposium 09: ATP and adenosine receptors and roles

S0902
Extracellular adenosine generation and signaling in cardiovascular, pulmonary and renal
injury
ELTZSCHIG HK
Mucosal Inflammation Program, Department of Anesthesiology and Perioperative Medicine,
Colorado Denver, Denver, CO, USA

Extracellular adenosine has been implicated as endogenous distress signal modulating tissue
adaptation and repair. Particularly during conditions of limited oxygen availability (hypoxia,
ischemia), extracellular adenosine plays a critical role in adaptive responses. Here, we will review
recent studies utilizing genetic models of extracellular adenosine generation and signaling that
address the role of adenosine under such conditions. These studies include adenosine generation and
signaling during ventilator induced lung injury (Eckle et al. 2007), ischemic preconditioning of the
heart (Eckle et al. 2008) or the kidneys (Grenz et al. 2008). 1. Eckle, T., L. Fullbier, M. Wehrmann,
J. Khoury, M. Mittelbronn, J. Ibla, P. Rosenberger, and H.K. Eltzschig. 2007. J Immunol 178:8127-
8137. 2. Eckle, T., D. Kohler, R. Lehmann, K.C. El Kasmi, and H.K. Eltzschig. 2008. Circulation
118:166-175. 3. Grenz, A., H. Osswald, T. Eckle, D. Yang, H. Zhang, Z.V. Tran, K. Klingel, K.
Ravid, and H.K. Eltzschig. 2008. PLoS Medicine 5:e137.

S0903
Adenosine as regulator of sleep and wakefulness
PORKKA-HEISKANEN T
Institute of Biomedicine, University of Helsinki, Finland

Adenosine is an inhibitory neuromodulator with tight connection to energy metabolism: extracellular
adenosine concentration increases when energy balance in cells turns unfavorable. It has been
hypothesized that adenosine is one of the key molecules regulating sleep. Several experiments
provide evidence to support this hypothesis. Administration of adenosine or its agonists increase
sleep while administration of adenosine antagonists has the opposite effect. Importantly,
extracellular adenosine concentration increases is the basal forebrain (BF) during prolonged
wakefulness and decreases, when sleep after the wakefulness period is initiated. Blocking the
adenosine A1 receptors in the BF during the prolonged wakefulness period will prevent the
development of excessive sleep (=recovery sleep), that normally follows prolonged waking,
suggesting that, indeed, the increase in adenosine during prolonged wakefulness in one of the key
mechanisms of sleep homeostasis. The adenosine response appears to be connected to BF
cholinergic cells since specific lesion of these cells abolishes both the increase in extracellular
adenosine during prolonged wakefulness and the recovery sleep, introducing further evidence to
support adenosine‟s role as a regulator of sleep homeostasis. An interesting question, that so far
remains unanswered, is the origin of adenosine during prolonged wakefulness: is it produced by
astrocytes or (cholinergic) neurons?




                                                  55
                                              Abstracts

S0904
Knocking out the adenosine A1 receptor increases lipolysis and insulin release
FREDHOLM BB, JOHANSSON SM, GRAPENGIESSER E, HELLMAN B, KATZ A, SALEH A.
Department of Physiology and Pharmacology, Karolinska Institutet, Department of Medical Cell
Biology, University of Uppsala, Biomedicum Box 571, Uppsala, Department of Clinical Science,
CRC (UMAS), University of Lund, Malmö, Sweden

Adenosine is an ubiquitous autacoid that exerts its effects on four G protein-coupled receptors, A1,
A2A, A2B and A3. Although adenosine is generally perceived as an emergency signal it plays also
some roles in basic physiology. This is partly evidenced by the fact that the adenosine receptor
antagonist caffeine has clear effects on e.g. wakefulness and renal function already under basal
conditions. We have used mice that lack adenosine A1 receptors (A1R-/-) to probe the roles of this
receptor in the regulation of intermediary metabolism. Fat cells from A1R(-/-) mice showed a higher
basal and stimulated lipolysis and cAMP production, but did not show any response to adenosine
analogues, adenosine deaminase or methyl xanthines. Fat cells from A1R (+/-) mice, which have
half the number of receptors, required twice as high dose of adenosine for any given antilipolytic
effect. There were no compensatory increases in the potency of nicotinic acid, prostaglandin E2 or
insulin as antilipolytic agents in A1R (-/-) adipocytes. Insulin release from pancreas was increased in
A1R (-/-) mice. In particular pulsatile insulin release during the later phase of secretion was
markedly influenced. Insulin-stimulated glucose uptake was slightly but not significantly reduced in
A1R (-/-) muscle and adipocytes. Glucose tolerance was unaltered in young fed mice. Thus,
adenosine, via A1 receptors play a physiological role in regulation of metabolism. Adenosine,
glucose tolerance, adipose tissue, pancreas.




                                                  56
                                               Abstracts


                              Symposium 10: Senses at the threshold

S1001
Vision at extreme
GOVARDOVSKII VI
Institute for Evolutionary Physiology & Biochemistry, Russian Academy of Sciences, 194223 St.
Petersburg, Russia

The basic question is, what factors, physical, biochemical and physiological, set the limits of visual
capabilities in the intensity (sensitivity), spatial (acuity), and spectral (wavelength) domains, and
how are they related to each other? There is a trade-off between sensitivity, visual acuity and color
vision that roots in the quantum nature of light. A certain minimum number of quanta is required to
paint a picture with a given sharpness and smooth shades of color. Thus low-light vision can only be
improved by collecting more photons, that is by making eyes bigger, increasing the fraction of
absorbed light, and adjusting spectral sensitivity of photoreceptors to the spectrum of ambient
illumination. However, quantum-limited performance can further be spoiled by the noise originating
in the visual system. One of the noise sources is the thermal activation of visual pigment molecules.
It results in spontaneous excitation events that occur at random intervals and are identical to those
signaling real photon absorptions. They constitute an irreducible noise that is added to the quantum
noise of the image itself. Both experimental and theoretical studies show that the visual pigment
noise steeply increases with shifting the spectral sensitivity curves towards longer wavelengths. This
explains the occurrence of a Purkinje shift during rod- cone transition, and sets the "red" limit of the
visible spectrum. Maximizing signal-to-noise ratio can be an important factor that governs the
selection of an optimum visual pigment for a given photic environment.

S1002
Colour vision limits in dim light
KELBER A, ROTH LSV, LIND O
Department of Cell and Organism Biology, Vision Group, Lund University, Helgonavägen 3, S-
22362 Lund, Sweden

Nocturnal vision is limited by the low numbers of photons available. To improve the accordingly
bad signal-to-noise ratios, nocturnal vision often involves of spatial, temporal and spectral pooling.
In humans, spectral pooling – pure rod vision resulting in nocturnal colour-blindness – occurs in
light levels dimmer than a half moon. Nocturnal colour vision is useful because the colour of light
changes dramatically between dim twilight and moon- or starlight. The changes of the illumination
colour make achromatic vision much less reliable compared to colour vision. However, the absolute
threshold of colour vision has not been investigated in many animals so far. Using behavioural
methods, we found that horses, with the largest terrestrial mammal eyes, have a very similar colour
vision threshold as humans, and that diurnal birds loose colour vision at brighter light levels then we
do. But we have also shown that nocturnal hawkmoths, nocturnal carpenter bees and nocturnal
geckos can use colour vision in very dim light when humans are colour-blind. Besides these three
groups, for which colour vision has been proven behaviourally, other animals may have nocturnal or
dim light colour vision. Possible candidates include toads and frogs with two rod types, other large
nocturnal insects, spiders and some deep-sea fishes. Finally, we are also studying, how nocturnal
animals switch to other sensory modalities when vision becomes less reliable. Using moths as


                                                   57
                                               Abstracts

models, we find that while diurnal species rely on colour in the first place, nocturnal species rely
more strongly on olfactory cues.

S1003
Mammalian hearing sensitivity in air, and in water
NUMMELA S
University of Helsinki, Department of Biological and Environmental Sciences, POB 65 (Viikinkaari
1), 00014 Helsinki, Finland

For sound energy to reach the inner ear, an impedance matching device is needed between the
surrounding medium and the inner ear fluid. The mammalian middle ear performs two tasks:
impedance matching and sound amplification. For terrestrial mammals, the middle ear ossicular
inertia is the main factor limiting the high- frequency hearing, and for animals ranging from bats to
elephants, the high-frequency hearing limit (HFHL) is inversely proportional to the cubic root of the
ossicular mass (Hemilä et al. 1995). The middle ear size grows with animal size, and for smallest
mammals able to hear very high frequencies, additional constraints are posed by the cochlear
sensitivity. The co- existence of inertial and cochlear constraints is seen among pinnipeds where the
ossicular inertia in air, and cochlear sensitivity in water are the limiting factors for phocids, but the
cochlear sensitivity alone limits hearing in both media among otariids (Hemilä et al. 2006). The
origin of cetaceans is one of the best examples of macroevolutionary changes in life‟s history and
shows how the hearing organ changed while whales became obligately marine (Nummela et al.
2004). Due to different acoustics of air and water, sound transmission from the surrounding water to
the inner ear demands a reversed impedance matching than is required on land. This kind of
mechanism has evolved in odontocetes, and despite a much larger ear size can transmit equally high
frequencies as small bats hear in air. Additionally, the sensitivity is 50-100 times better than in man.
Hemilä, S., Nummela, S. & Reuter, T. 1995. Hear Res 85, 31-44. Hemilä, S., Nummela, S., Berta, A.
& Reuter, T. 2006. J Acoust Soc Am 120, 3463-3466. Nummela, S., Thewissen, J.G.M., Bajpai, S.,
Hussain, S.T. & Kumar, K. 2004. Nature 430, 776- 778.

S1004
Trade-offs in visual detection
DONNER K
Department of Biological and Environmental Sciences, P.O. Box 65 (Viikinkaari 1), FI-00014
University of Helsinki, Finland

Limits to sensory detection are always at some (physical or neural) level set by signal/noise. In
vision, photon statistics sets one absolute limit to the information that can be extracted from an
image, but the information may be partitioned differently depending on the needs of an animal in a
particular situation. Primarily, a balance has to be struck between sensitivity and resolution,
secondarily between different kinds of resolution: spatial, temporal and chromatic, with polarization
as an additional dimension in some species. Realizable combinations of these depend on the
anatomical and physiological implementations in each species. As an example, I shall summarize
our studies on temporal integration (which favours sensitivity) vs. temporal resolution in humans
and amphibians. In rod vision, the balance is set at the very input: the time scale of rod
phototransduction limits all temporal aspects of scotopic vision. Cone vision, however, appears to be
slower than cone phototransduction would allow. Modelling suggests that “unnecessarily” fast cone

                                                   58
                                              Abstracts

signals in humans may be useful for intra-retinal signal processing that serves to enhance spatial
resolution under small eye movements.




                                                  59
                                              Abstracts


              Symposium 11: Long-term blood pressure regulation in normal man:
                           New aspects of neuro-humoral control

S1101
Proximal sodium reabsorption: a role in the regulation of blood pressure?
BURNIER M
Service of Nephrology and Hypertension Department of Medicine, University Hospital, Lausanne,
Switzerland

The molecular characterization of rare monogenic salt-sensitive forms of hypertension has focused
the attention on alterations of renal sodium handling occurring essentially in the distal nephron
where sodium excretion is finely tuned to maintain sodium balance. These observations however
only partly explain the mechanisms of the blood pressure response to changes in sodium intake
observed in patients with essentially hypertension. In fact, there is growing experimental, clinical,
genetic, and epidemiological evidence that proximal tubular sodium reabsorption is also an
important determinant of blood pressure control in humans. Thus, we have observed important
differences in renal tubular sodium handling and particularly in proximal sodium reabsorption
depending on race and on the genetic background Considering the actual level of evidence,
segmental renal sodium handling should perhaps be investigated more frequently in the future. But
to generate a greater interest for this type of research, new techniques should be developed enabling
to assess reliably the renal tubular response to sodium in humans. A better characterization of the
molecular mechanisms whereby renal tubules adapt to changes in sodium intake would certainly
improve our understanding of the role of the kidney in mediating salt sensitivity in hypertension and
diseases such as obesity and diabetes.

S1102
Sodium intake, renal sodium reabsorption and genetic predisposition in long term blood
pressure control
MELANDER O
Department of Clinical Sciences-Hypertension and Cardiovascular disease, Lund University,
Malmö, Sweden

Industrially processed food contributes with 75 % of total daily salt intake and recent long term trials
controlling the actual salt intake by providing all foods and drinks for study participants have shown
that the mean blood pressure lowering effect of a 6 gram reduction of salt intake is approximately
6/3 mmHg, i.e. an effect comparable to antihypertensive pharmacological monotherapy. Importantly,
whereas most individuals are salt sensitive, the degree of salt sensitivity (the degree of blood
pressure reduction following a lowering of salt intake) differs between individuals. A low plasma
concentration of renin predicts high salt sensitivity. This suggests that enhanced renal tubular sodium
reabsorption, which suppresses renin release from the juxtaglomerular apparatus, contributes to high
degree of salt sensitivity. Monogenic rare forms of human hypertension are predominantly caused by
mutations leading to increased tubular sodium reabsorption through the epithelial amiloride sensitive
sodium channel (ENaC) which leads to suppression of renin release and severe salt sensitive
hypertension. Whereas common forms of hypertension and population blood pressure variation is
not influenced by genetic variation of ENaC, common genetic variation of the key regulatory protein
of ENaC, the Neural Precursor Cell Expressed Developmentally Down-regulated 4-Like (NEDD4L)


                                                  60
                                              Abstracts

is associated with higher population blood pressure, increased salt sensitivity, reduced plasma renin
concentration and increased risk of cardiovascular disease. Based on recent results, this talk will
review the effects of salt intake on blood pressure as well as determinants of salt sensitivity, blood
pressure and its long term cardiovascular consequences in man.

S1103
Abstract missing

S1104
Abstract missing




                                                  61
                                                Abstracts


           Symposium 12: The effects of sleep loss on performance and human health

S1201
Sleep loss and accident risk
ÅKERSTEDT T
Stockholm University and Karolinska Institutet, Stockholm, Sweden

The brain responds metabolically to reduced/impaired sleep, time spent awake, and time of day
(circadian low). All will separately, and particularly together, cause alertness to vary from very high
levels down to extreme sleepiness. This will strongly affect performance capacity and safety. Fatigue
is now, for example, a more important factor behind road accidents than alcohol and the interaction
between the two potentiates their individual risk values several times over. This is mainly seen in
transport work but also occurs in industry and health care. Nights on call (for the MD) are related to
significantly increased risk for the patients. The high risk also means that the interest in
countermeasures has increased. The best ones appear to be taking a nap or consuming caffeine
(blocking adenosine receptors in the hypothalamus which should respond to sleep loss, time awake,
etc). Another countermeasure is alertness monitors that warn when sleepiness reaches dangerous
levels. A number of European and US projects have been focus on the identification of sensors and
on the building of efficient warning systems. The most sensitive variables seem to be eye blink
duration or derivatives thereof, or lateral variability of lane position in vehicles. Traditional
measures, such as the EEG do detect sleepy driving but the sensitivity and specificity is modest. A
final type of countermeasure involves fatigue risk management. It has recently come into focus in air
transport but is making headway also in road transport and industry. It essentially involves pre-work
evaluation of the schedule using some sort of alertness prediction model, training personnel in
fatigue risk management and regular monitoring of fatigue through a routine reporting system. In
most cases individuals are aware of the current level of sleepiness. However, once the individual has
reached a high level of sleepiness there will be no further signal that the limit has been reached and
the persistent individual may simply decide to take the chance and trust his/her ability to fight
sleepiness. This lack of insight is probably behind many sleep related accidents and the person that
causes this type of accident may refuse to admit to any serious sleepiness and blame a ”black-out”. A
truck driver might blame a non-existent moose for his driving off the road. This would in most
countries cause an acquittal since the legislation only recognizes guilt if the defendant was aware of
his level of sleepiness. Thus, one needs to change the legislation to refer to illegal levels of sleep loss
(≤5h) rather than to perceived sleepiness, analogous to drunken driving sentences being based on
blood alcohol levels rather than degree of intoxication.

S1202
Sleep loss and immune function
PORKKA-HEISKANEN T
Institute of Biomedicine, Helsinki, Finland

Restricting sleep, either voluntarily or as response to work requirements, is increasing in all western
societies. Homeostatic sleep regulation responds to this restriction by increasing the duration and
depth of the sleep period that follows the prolonged wakefulness period. Occasional prolongation of
wakefulness can be thus be compensated for. However, when sleep restriction becomes chronicle,
several symptoms of restricted performance as well as health consequences start to appear.


                                                    62
                                               Abstracts

Immune system is one of the first to react to prolonged wakefulness. Several components of the host
defense reaction are activated, including the folded protein response, as indicated e.g. by increase in
BiP expression, iNOS synthesis and in humans, by increase in c-reactive protein, CRP. In addition,
several cytokines respond to sleep restriction. Interestingly, many cytokines are also sleep-inducing,
giving an explanation to the well-known phenomenon that during infection both animals and humans
increase sleep. If sleep restriction continues, the immune system starts to loose its effectiveness: e.g.
wound healing is compromised and the ability of young volunteers with partial, chronicle sleep
restriction produced less antibodies as response to influenza vaccination than those who had slept
normally. In extreme cases (experiments done with rodents), lethal infection from normally non-
pathogenic bacterial flora takes place.

S1203
Cognitive performance during sleep loss: What are we measuring?
HAAVISTO M-L
Finnish Institute of Occupational Health, Brain and Work Research Centre, Helsinki, Finland

Cognitive performance degrades selectively under sleep loss. Thus, it appears that specific demands
associated with given cognitive task influence the performance results under sleep loss. However,
most of the studies that examine the effects of sleep deprivation on cognitive performance have
focused on measures that are sensitive to sleepiness, such as simple reaction time, but have little to
do with the true nature of cognitive demands in real work performance. Some other studies have
reported that sleep loss does not lead to detrimental effects on higher cortical functioning, as
measured with neuropsychological tests that have been mainly developed for detecting different
disorders. But in these studies participants have been young and screened for not having any
neurological disorders. In this presentation I will focus on human cognitive processes and some
other critical factors for selecting cognitive measures to evaluate the effects of sleep loss on
performance. The study of five days partial sleep loss and following two days with 8 hours sleep will
be presented as an example. In this study, cognitive performance has been measured repeatedly in
multitasking, the declarative and procedural memory tasks and the working memory tasks in each
day of the experiment. The cognitive performance declined selectively in multitasking and
procedural memory performance, while declarative and working memory performance were very
little affected by sleep loss. In sum, the more systematic studies evaluating the effects of sleep loss
on human cognitive processes are needed.

S1204
Sleep, mood and genes
PAUNIO T
Departmernt of Psychiatry, Helsinki University Hospital, Helsinki, Finland

Insomnia is a well-recognised risk for depression but it is not well known if depression induces
insomnia in turn, or if there were genetic factors in common for both features. We investigated the
direction of causality between problems in sleep and depressed mood, and also whether genes in
common underlie both traits, in a nation-wide cohort sample of 18 631 same-sex twins with a
follow-up interval of six years. As an approximation for depressed mood, we used information on
life dissatisfaction that has previously been found to be strongly associated to symptoms of
depression and with high specificity and sensitivity to the disorder itself (Paunio et al. submitted).

                                                   63
                                               Abstracts

Most individuals (59 %) with new-onset life dissatisfaction had experienced suboptimal sleep at the
baseline. Poor sleep predicted life dissatisfaction in a consistent pattern (OR=2.1 from logistic and
3.0 from conditional logistic regression analysis), but life dissatisfaction did not predict poor sleep.
There was substantially heritability for both traits, but their shared genetic component was relatively
weak (genetic correlation of .21 for males and .27 for females in a multivariate genetic model).
These results suggest that most of the association between symptoms of insomnia and depressive
disorder is due to an effect of poor sleep on mood. Majority of the contributing genetic factors were
unique, consistent with the hypothesis that poor sleep has a neurophysiological effect on brain,
emotions and mood. Finally, revelation of genetic factors contributing to experiences of poor sleep
will bring new insight into genetic background of depressive disorder as well.
Paunio, T., Korhonen, T., Hublin, C., Partinen, M., Kivimäki, M, Koskenvuo, M, Kaprio, J.
Manuscript submitted




                                                   64
                                             Abstracts


                Symposium 13: Oxygen-dependent regulation of gene expression

S1301
Molecular imaging of cellular oxygen sensing
FANDREY J
Institut für Physiologie, Universität Duisburg-Essen, Germany

Dissection of the hypoxia induced transcription factor complex bound to the erythropoietin enhancer
under hypoxic conditions has led to the identification of Hypoxia-inducible factor-1 (HIF-1). From
here the whole cascade of cellular O2 sensing was followed back ultimately leading to the
elucidation of cellular O2 sensors, the prolyl and asparagyl hydroxylases that regulate abundance and
activity of oxygen-regulated -subunit of HIF-1. Assembly of the HIF- 1 complex then requires
dimerization with constitutive nuclear -subunit. Mobility studies of fluorescently labelled HIF-1
subunits by fluorescence recovery after photo bleaching (FRAP) revealed that HIF-1 migrates
more slowly than HIF-1 within the nucleus indicating that both subunits do not immediately “find”
each other but may be prone to modification prior to dimerization. To localize HIF-1 subunits within
the nucleus of hypoxic cells we applied 2-photon-laser microscopy (2PLM). HIF-1 assembly was
then studied in living cells in a specialized hypoxic chamber mounted on the microscopic stage
which allows the in vivo analysis under a well defined oxygen tension. Assembly of the HIF-1
complex was analysed by fluorescence resonance energy transfer (FRET) and determined the
distance of - and -subunits of the transcriptionally active HIF-1 complex bound to DNA.
Interestingly, the N- and C-terminus of HIF- 1 and - were closely co- localized. This organization
of the HIF complex is different from so far described two dimensional models but is in line with the
very recent structural consideration that heterodimer formation of the HIF complex relies on anti-
parallel association of the PAS B domains within the - and -subunit.

S1302
Abstract missing

S1303
Abstract missing




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                                             Abstracts


          Symposium 14: The role of nitric oxide in the development of hypertension

S1401
Role of NO deficiency in salt-sensitive hypertension
CARLSTRÖM M, SÄLLSTRÖM J, LAI E, PATZAK A, BROWN RD, WÅHLIN N, PERSSON
AEG
Department of Medical Cell Biology, Division of Integrative Physiology, Uppsala University,
Sweden

BACKGROUND: The kidneys play a key role in the homeostatic regulation of body fluid status,
and an impaired renal function has been demonstrated in salt-sensitive hypertension. During the last
years it has become evident that NO systems play an important role in blood pressure regulation and
that NO deficiency may in fact be involved in the development of hypertension. To investigate the
mechanisms for this worldwide growing disorder we have used different experimental models: 1)
Chronic blockade of nNOS in rats 2) Hydronephrosis induced by chronic partial ureteral obstruction
3) Reduction in nephron number after completed nephrogenesis 4) Genetic modified mice (nNOS-
deficient, SOD1- deficient, SOD1-transgenic) METHODS: Renal function in terms of blood flow,
glomerular filtration rate, electrolyte excretion has been investigated. The tubuloglomerular feedback
(TGF) mechanism was investigated by stop-flow pressure technique. Blood pressure and heart rate
was measured both acutely and with telemetry, during different sodium conditions (i.e. low, normal
and high NaCl-diet). Renal microcirculation was investigated by studying the responsiveness in
isolated and perfused afferent arterioles of mice. RESULTS: In our models, we have found that
reduced renal NO availability and subsequent resetting of the TGF-response plays an important role
for the development of salt-sensitive hypertension. Oxidative stress (by superoxide) can enhance the
TGF responsiveness, both directly by constricting afferent arterioles and indirectly by scavenging
NO, and thereby play an important role in the development and maintenance of hypertension.
Treatment modalities that increase NO or decrease oxidative stress can attenuate the salt- sensitive
hypertension. In this presentation the results of our studies will be discussed.

S1402
Mechanisms of NO release in angiotensin II induced hypertension
IVERSEN BM
Renal Research Group, Institute of Medicine, University of Bergen, Norway

Mechanisms of NO release in angiotensin II induced hypertension BJARNE M. IVERSEN, Renal
Research Group, University of Bergen, Norway. 2K1C is a model of renovascular hypertension with
an exaggerated intracellular calcium (Ca2+i) response to angiotensin II (ANG II) in isolated afferent
arterioles (AAs) from the clipped kidney. To explore the role of NO release in this model, we
studied ANG II (10-7 mol/L) induced calcium signalling and contractility with L-NAME. In AAs
from the non-clipped kidney, L-NAME increased the ANG II induced Ca2+i response from 0.28±0.05
to 0.55±0.09 (Fura-II 340 nm/380 nm ratio), and increased contraction from 80±6 % of baseline to
60±6 % of baseline (p < 0.05). In vessels from sham and clipped kidneys, L-NAME had no effect. In
DAF-FM loaded AAs from the non- clipped kidney, ANG II increased NO-derived fluorescence to
145±34 % of baseline (p < 0.05 vs. sham), but not in sham or clipped kidney vessels. In the non-
clipped kidney, cationic aminoacid transferase (CAT) 1 and 2 mRNA was increased in both kidneys
from 2K1C (p < 0.05), indicating an increased transport-capacity of the NO precursor L-arginine. To


                                                 66
                                               Abstracts

pursue the role of ANG II, we used ANG II infused rats using Alzet micropumps for 14 days. Ten
animals from each group received the AngII AT1 receptor blocker Losartan in the drinking water.
mRNA expression was investigated for the genes for eNOS, CAT-1, CAT-2, DDAH-1, DDAH-2,
Arginase-1 and Arginase-2 using real time reverse transcriptase PCR. 18S RNA was used as internal
control. eNOS, DDAH-1 and Arginase-2 were not changed by ANG II infusion and were not
affected by Losartan treatment. CAT-2, DDAH-2 and Arginase-1 were significantly increased in
AngII infused rats and the expressions were blunted by Losartan treatment. In conclusion, the
mRNA expressions of CAT-2, DDAH-2 and Arginase-1 AAs seem to be regulated by AT1 receptor
dependant mechanism.

S1403
The role of the nitrate-nitrite-nitric oxide pathway in vascular control
PETERSSON J
Department of Medical Cell Biology, Division of Integrative Physiology, Uppsala University,
Uppsala, Sweden

Nitric oxide (NO), generated by nitric oxide synthases, is a key regulator of vascular tone. Recently,
a fundamentally different pathway for NO production was described in which nitrate and nitrite are
converted to NO and other bioactive nitrogen oxides. Research conducted during the last years has
changed our way of viewing these anions, since nitrate and nitrite were long known predominantly
as inert oxidative end products of NO metabolism. Plasma nitrate and nitrite do not only reflect the
endogenous NOS activity since dietary sources of nitrate have a major impact of the circulating
levels of these anions. Nitrate is abundant in our diet, and particularly high levels are found in many
vegetables. After ingestion and absorption, circulating nitrate is actively taken up and concentrated
in the salivary glands. When the nitrate-rich saliva is secreted into the oral cavity, nitrate reducing
bacteria on the tongue reduce some of the nitrate to nitrite. Therefore, intake of nitrate-rich
vegetables results in elevated plasma levels of both nitrate and nitrite. More recent, focus has been
directed towards the role of nitrate intake and plasma nitrite in the control of blood pressure. In fact,
there are now results showing that acute nitrate intake reduces blood pressure in humans, and also
that nitrate intake over a longer period lowers blood pressure in animals. A diet rich in fruits and
vegetables has been shown to reduce blood pressure and increased nitrate intake may be one of the
explanations for this. Another fascinating aspect is revealed, since humans are dependent on the oral
microflora for conversion of nitrate to nitrite, highlighting the importance of symbiosis between us
and our microbes.

S1404
Cyclic AMP-mediated activation of eNOS by vasodilator agonists; role in blood pressure
regulation
JENSEN BL, HRISTOVSKA A-M, RASMUSSEN LE, HANSEN PB, NIELSEN SS, NÜSING
RM, NARUMIYA S, VANHOUTTE P, SKØTT O
Department of Physiology and Pharmacology, Institute of Medical Biology, University of Southern
Denmark, J.B. Winslowsvej 21, DK-5000 Odense C, Denmark

We have examined mechanisms for PGE2-mediated vasodilatation and hypothesized an involvement
of endothelial NO synthase (eNOS). Aortic rings from mice were used for contraction studies. Blood
pressure changes in response to PGE2 were measured in conscious mice. Single doses of PGE2

                                                   67
                                            Abstracts

caused concentration-dependent relaxations during contractions to phenylephrine (EC50 =5 × 10- 8
mol/L). Relaxation after PGE2 was absent in rings without endothelium, in rings from eNOS-/- mice
and was abolished by L-NAME and by inhibition of guanylate cyclase. Vascular cGMP, but not
cAMP, content increased after PGE2. PGE2- induced relaxations were abolished by the EP4
receptor antagonist AE3-208 (10-8 mol/L) and mimicked by an EP4 agonist (AE1-329, 10-7 mol/L)
in the presence of endothelium and eNOS only. Relaxation was similar to control in rings from EP2-
/-. Inhibitors of the cAMP-PKA pathway attenuated, while the inhibitor of protein phosphatase 1C,
calyculin (10-8 mol/L), abolished the PGE2-mediated relaxation. In aortic rings, PGE2
dephosphorylated eNOS at Thr.495 Chronically catheterized eNOS-/- mice were hypertensive (137
mmHg±3.6, n = 13 vs. 101 mmHg±3.9, n = 9) and exhibited a lower sensitivity of blood pressure
reduction in response to PGE2 compared to wild type mice. There was no difference in the blood
pressure response to nifedipine. These findings show that PGE2 elicits EP4 receptor-mediated,
endothelium- dependent stimulation of eNOS activity by dephosphorylation at Thr495 resulting in
guanylyl cyclase-dependent vasorelaxation and accumulation of cGMP in aortic rings. We speculate
that activation of eNOS and NO release may be a common feature of agonists that elicit cAMP
formation in the endothelium.




                                               68
                                              Abstracts


                     Symposium 15: Calcium signaling in heart and muscle

S1501
Cardiac development, dysrhythmia, inotropy and hypertrophy; the many possible roles of
inositol 1,4,5-trisphosphate in the heart
BOOTMAN MD, HARZHEIM D, FEARNLEY C, HIGAZI D, RODERICK HL
Laboratory of Molecular Signalling, The Babraham Institute, Babraham, Cambridge, CB22 3AT UK

The role of inositol 1,4,5-trisphosphate (InsP3) in cardiac myocyte function is unclear and
controversial. Although agonists that activate InsP3 generation are positive inotropic agents in the
heart and have been implicated in various cardiac pathologies, it is unclear whether InsP3 is
responsible for any of their effects. We have investigated the expression and function of InsP3
receptors (InsP3Rs) in neonatal rat ventricular myocytes, and adult rat ventricular and atrial cardiac
myocytes. We find significant expression of InsP3Rs in each of these tissues, and can evoke
responses consistent with the receptors being functional and having access to replete calcium stores.
Significantly, the promiscuous calcium release from InsP3Rs means that they contribute
significantly to spontaneous diastolic calcium transients in hormone-stimulated ventricular and atrial
myocytes. In recent studies, we have been examining the putative contribution of InsP3Rs to
hypertrophic growth. These experiments were performed using spontaneously contracting
monolayers of primary cultures of neonatal ventricular rat myocytes (NRVM). NRVMs were
prepared by collagenase digestion of cardiac ventricles isolated from 1-2 day old Wistar rat pups.
Quantitation of atrial natriuretic factor (ANF) by immunofluorescence and real-time PCR, together
with cell surface area were used as measures of hypertrophy. Our data demonstrate that InsP3-
induced calcium release is required for the induction of hypertrophy downstream of Gq- coupled
receptors or increased workload. Furthermore, InsP3-induced calcium release may represent a
mechanism by which gene transcription can be isolated from the increases in cytosolic calcium that
occur during every heart-beat.

S1502
Contribution of sarcoplasmic reticulum to excitation-contraction coupling of the fish heart
VORNANEN M, HAKKO H, HAVERINEN J
University of Joensuu, Faculty of Biosciences, PO Box 111, 80101 Joensuu, Finland

In mammalian cardiac myocytes, a small entry of extracellular Ca triggers a much larger release of
Ca from the sarcoplasmic reticulum (SR) by a process called Ca-induced Ca release. Contraction of
the fish heart is only weakly or moderately sensitive to ryanodine, a specific blocker of the SR,
suggesting that SR Ca stores provide relatively little Ca for contraction in fish cardiac myocytes. To
this end, Ca storing capacity and calsequestrin (CASQ) content of the fish cardiac SR were
determined by caffeine-induced Ca releases in single myocytes and immunoblotting of cardiac
proteins with an antibody specific to the fish cardiac CASQ, respectively. The Ca storing capacity of
the fish cardiac SR is in many fish species larger than that of the mammalian SR and the Ca-
buffering protein, CASQ, is abundantly expressed in the fish hearts. Thus, the apparently small
contribution of SR Ca to contractile activation is not limited by the large SR Ca stores, but sooner
must be due to the release mechanism. One contributing factor to the weak release seems to be the
low Ca-sensitivity of the fish cardiac ryanodine receptors. On the other hand, the usage of SR Ca



                                                  69
                                               Abstracts

stores in excitation-contraction coupling is enhanced by acclimation to low temperatures, suggesting
that SR Ca stores are physiologically significant under changing temperature regimes.

S1503
Changes in skeletal muscle calcium handling during fatigue and recovery
WESTERBLAD H
Department of Physiology and Pharmacology, Karolinska Institutet, 171 77 Stockholm, Sweden

Acute exercise results in impaired muscle function; that is, fatigue develops. The impaired muscle
function in fatigue may be due to central (in the central nervous system) and/or peripheral (within
the muscle) factors. My research group studies mechanisms of peripheral fatigue with a special
interest in changes in sarcoplasmic reticulum (SR) calcium handling. Experiments are performed on
intact single muscle fibers obtained from mice and rats. Central fatigue is often assessed with the
twitch interpolation technique, where electrical stimulation is superimposed on an ongoing voluntary
contraction and the resulting change in force is measured. We have mimicked this stimulation
pattern in single fiber experiments and observe a force increase that could be interpreted as central
fatigue; the underlying principles will be discussed. Force recovery after fatiguing stimulation may
be slow, especially at low stimulation frequencies. Recent data from our laboratory show that two
different mechanisms can cause this prolonged force depression: decreased SR calcium release and
reduced myofibrillar calcium sensitivity. The relative importance of these two mechanisms appears
to depend on the production of different reactive oxygen species.

S1504
Metabolic modulation of skeletal muscle Ca2+ handling and excitability
ØRTENBLAD N
Institute of Sports Science and Clinical Biomechanics, University of Southern Denmark, Odense,
Denmark

Skeletal muscle energy turnover can increase more than 100 fold during high intensity exercise.
Thus, it is vitally important for muscle cells to possess signalling systems that keep the balance
between ATP-utilisation and ATP-production. However, little is know about the mechanism
whereby the muscle energy level or ATP-generating capacity restrains the rate of ATP- utilisation.
Here is presented such a system where events in the excitation-contraction (EC) coupling are
affected by glycogen content and localisation. We used a mechanically-skinned muscle fibre
preparation where the fibre structural integrity and excitability is maintained, and one can keep the
energy level (ATP and PCr) high and constant while varying glycogen levels. Data from this
preparation strongly indicates that glycogen has a structural, non-metabolic, role in maintaining
normal sarcoplasmic reticulum (SR) Ca2+ regulation in mammalian skeletal muscle. Further,
measurements of SR function following experimentally manipulation of muscle glycogen at both the
isolated muscle and human whole body level, strongly supports that glycogen has a structural role in
maintaining normal EC coupling in mammalian skeletal muscle, by modulating SR Ca2+ regulation.
In addition, we have shown that skeletal muscle force depression due to ionic perturbations, and
subsequent reduced SR Ca2+ release, is affected by muscle energy status (Macdonald et al. 2007).
Further, in isolated muscles and single fibres, loss of excitability can be partly reversed by the lactate
ion by closing Cl-channels. In conclusion, these findings illustrate how energy level modulates



                                                   70
                                           Abstracts

skeletal muscle EC coupling by affecting SR Ca2+ handling and excitability. Macdonald, W.A.,
Ørtenblad, N. & Nielsen, O.B. 2007. Am J Physiol 292, E771-E778.




                                               71
                                              Abstracts


                          Symposium 16: Ecophysiology and energetics

S1601
Metabolic senescence in a small passerine bird, the zebra finch
BECH C1, RØNNING B1, VERHULST S2, NOREEN E1, BERNTSEN HH1, MOE B1,3
1
  Department of Biology, Norwegian Universitgy of Science and Technology, NO-7491 Trondheim,
Norway. 2Behavioral Biology Group, University of Groningen, NL-9751 NN Haren, The
Netherlands. 3Norwegian Institute for Nature Research, Trondheim, Norway

Senescence involves a decline in physiological performance with age paralleled by increased
mortality and decreased reproductive rate. According to the “free radical theory of living” the main
cause of senescence, and eventually death, is the accumulated damage caused by reactive oxygen
species (ROS) and other free radicals on cell structures. The cellular production of ROS occurs in
itself in direct proportion to the aerobic processes, creating a possible mechanistic link between
aging and oxygen consumption. Studies on the relationship between age and metabolic rate have,
however, given conflicting results. The basal metabolic rate (BMR) is the lowest sustainable rate of
oxidative processes of a resting endothermic organism, which do not use energy on thermoregulation
and digestive processes. If a continuous effect of ROS is present throughout life, a steady decrease in
BMR should be observed. This is, however, not always observed in endotherms, suggesting the
mobilization of an efficient antioxidant enzymatic system in some species. In a longitudinal study
we have investigated the relationship between BMR and age in captive zebra finches (Taeniopygia
guttata) up to seven years of age. This exceeds the normal life span of wild-living individuals. We
found a significant decline in BMR with age in both female and male finches. In addition, we found
that individuals with the highest BMR, also showed the largest relative decline in BMR. These
results support the free radical theory of living, as the individuals with the highest BMR would be
expected to produce more ROS, which in turn would result in more cellular damage. Our results also
indicate that these small passerine birds may have traded defense against ROS with energy use for
other physiological functions.

S1602
Coping with oxygen shortage - physiological adaptations to hypoxia in the deep-diving hooded
seal
FOLKOW LP
Department of Arctic Biology, University of Tromsø, Breivika, NO-9037 Tromsø, NORWAY

Seals may perform long-duration dives that leave them severely hypoxic, and thereby represent
interesting mammalian models for comparative studies of hypoxia tolerance, particularly in relation
to brain function. The arctic ice- breeding hooded seal (Cystophora cristata) may dive to ~1 000 m
for durations of >50 minutes (Folkow & Blix 1999), an ability that is related to its high capacity to
store oxygen in a large haemoglobin-rich blood volume and in skeletal muscles that carry the highest
myoglobin concentration yet recorded (Burns et al. 2007). Like other seals, hooded seals also display
distinct diving responses encompassing bradycardia and massive peripheral vasoconstriction that
limits the distribution of oxygenated blood to all but the most hypoxia- vulnerable tissues. A diving-
induced vascularly mediated body cooling causes brain temperature to drop by up to 3oC, thereby
depressing metabolism as well as offering protection against hypoxic and oxidative stress (Odden et
al. 1999). We have recently also found that hooded seal cortical neurons are intrinsically more


                                                  72
                                              Abstracts

hypoxia tolerant than cortical neurons of non-diving species, possibly due to enhanced anaerobic
capacity, channel arrest mechanisms and effects of an unusual distribution of the neurally based
respiratory protein neuroglobin. Lessons learned from studies of survival strategies in hypoxia-
tolerant mammals like hooded seals may potentially help guide efforts to find effective treatments of
hypoxic injury in man. Burns, J.M., Lestyk, K.C., Folkow, L.P., Hammill, M.O. & Blix, A.S. 2007.
J. Comp. Physiol. B. 177, 687-700. Folkow, L.P. & Blix, A.S. 1999. Polar Biol. 22, 61-74. Odden,
Å, Folkow, L.P., Caputa, M., Hotvedt, R. & Blix, A.S. 1999. Acta Physiol. Scand. 166, 77-78.

S1603
Comparative physiology of fatty acid mobilization and hepatic lipidosis
NIEMINEN P1,2, ROUVINEN-WATT K3, KÄKELÄ R4, MUSTONEN A-M2
1
  University of Oulu, Department of Biomedicine, Institute of Anatomy and Cell Biology, P.O. Box
5000, FI-90014 Oulu, 2University of Joensuu, Faculty of Biosciences, 3Nova Scotia Agricultural
College, Truro, Canada, 4University of Helsinki, Biomedicum

Previous studies on laboratory rodents, rabbits and humans have demonstrated that fatty acid (FA)
mobilization from white adipose tissue (WAT) is selective and its efficiency is related to FA
structure. The present studies explored whether similar selectivity of FA mobilization was
manifested in carnivores experiencing seasonal food scarcity and abundance. Fractional mobilization
from and incorporation into WAT of a wide spectrum of FA were studied by gas–liquid
chromatography from the subcutaneous WAT of several captive species of the order Carnivora
(American mink Neovison vison, European polecat Mustela putorius, American marten Martes
americana, sable Martes zibellina) and free-ranging raccoon dogs (Nyctereutes procyonoides). Due
to food deprivation, n-3 polyunsaturated FA (PUFA) decreased in proportion compared to n-6
PUFA. Mobilization correlated inversely with the FA chain length but increased with unsaturation
and when the first double bond was located closer to the methyl end. 18–20C n-3 PUFA and 14–17C
monounsaturated FA (MUFA) were preferentially mobilized while 19–24C saturated FA and MUFA
were preserved. In raccoon dogs, the wintertime FA mobilization was selective and mostly
confirmed the patterns of FA release in captivity during wintering. The summertime FA
incorporation correlated inversely with the chain length and increased with unsaturation and in
MUFA and PUFA with the double bond(s) closer to the methyl end. FA incorporation reversed the
wintertime losses of the preferably mobilized FA. The principles of selective FA mobilization were
valid in wild mammals. The loss of n-3 PUFA during food deprivation can be related to
development of hepatic lipidosis during fasting as examined in the European polecats. This can have
future applications in research concerning non-alcoholic fatty liver disease of humans caused by
obesity and rapid weight loss and considered to be the hepatic manifestation of the metabolic
syndrome.

S1604
The role of testosterone in reproductive physiology and evolution of a mammalian species
MOKKONEN M, KOSKELA E, MAPPES T, MILLS S
Department of Biological and Environmental Science, PO Box 35 (YA), FIN-40014, University of
Jyväskylä, Finland

Hormones have the capacity to alter the physiology and behaviour of animals. This has important
implications for an individual‟s life history, especially considering the central role of sex hormones

                                                  73
                                              Abstracts

in reproductive processes. Studies of the bank vole (Myodes glareolus) have revealed the effects of
testosterone (T) on sexual selection. Using offspring of wild-caught individuals, we artificially
selected for high and low T. We measured the effect of testosterone selection on life history traits,
including basal metabolic rate (BMR), reproductive success, litter measures as well as sperm
characters. Testosterone had a different effect on metabolism in males compared to females. An
antagonistic sex difference was also found in the way T affected reproductive success: high T males
and low T females were the most successful. Furthermore, there was evidence of an effect of T on
the siring ability of sperm based on sperm characteristics. Overall, we found that testosterone has a
potent effect on reproduction in this wild species. The role of testosterone in physiological processes,
reproductive behaviours and the evolution of this common European mammal is discussed.




                                                  74
                                              Abstracts


    Symposium 17: Orthostatic influence on cerebral blood flow - from humans to giraffes

S1701
Brain circulation in relation to upright/supine position in the normal and the injured brain
GRÄNDE P-O
Department of Anaesthesiology and Intensive Care, University of Lund, Sweden

The human brain is exposed to large variations in gravitational forces both on the arterial and the
venous side of its circulation when changing from supine to upright position and vice versa.
As the venous pressure outside the dura is clearly lower than the 10 mmHg inside the dura, this
pressure fall will create a passive venous collapse just inside the dura. The collapse will increase
upon an increase in intracranial pressure and decrease in venous pressure. A venous pressure fall just
outside the dura following a change from supine to upright position, therefore, will not be transferred
to the brain due to the compensatory passive increase in the venous collapse. A similar collapse of
the veins outside the dura will have similar protecting effect (1). The normal brain is protected from
arterial pressure variations via an active autoregulatory mechanism, which limits blood flow and
hydrostatic pressure variations upon variation in position. The autoregulatory mechanism can be
abolished following head injury. The venous protecting system presupposes a closed spinal canal. If
the distal spinal canal is opened, as sometimes occurs in clinical practice after spinal lumbar
puncture, the gravitational force on the venous side will be compensated for by a similar large force
of the spinal canal, and head elevation may dilate the venous vessels (no venous collapse). This will
increase the blood volume close to the pain sensitive meningeal structures. This scenario may result
in a violent headache appearing after changing from supine to upright position and disappearing
when returning to the supine position (2). 1) Kongstad L, Grände PO. Acta Anaesth Scand 1999;
43:501-8 2) Grände PO. Acta Anaesth Scand. 2005; 49:619-26

S1702
Orthostatic challenges of the giraffe’s cardiovascular system
BRØNDUM E
Institute of Physiology and Biophysics, University of Aarhus, Aarhus, Denmark

Due to its extraordinary height, the giraffe experiences great cardiovascular challenges both in the
upright position and when the head is lowered to drink. When assuming the drinking position, the
giraffes makes a ~4 m vertical change in head position within a second and arterial pressure at the
level of the head is expected to increases some 250 mmHg. This increase is considered to outrange
the cerebral autoregulation and should therefore, at least potentially, lead to cerebral edema and
eventual hemorrhage. After drinking (water) the giraffe is able to lift the head back to upright
position in one consecutive movement without showing signs of dizziness. We measured pressure
and flow along with vessels diameter and spinal fluid pressure in the neck of 5 anaesthetized
spontaneously breathing giraffes, suspended in upright position while changing the head position.
When the giraffes head was lowered to heart level, the mean arterial pressure (MAP) and central
venous pressure (CVP) decreased while spinal fluid pressure increased. The jugular vein was
distended and a total of ~1.2 l of blood accumulated in the veins when the head was lowered. When
the head was raised, the jugular vein collapsed and blood was returned to the central circulation,
restoring CVP and MAP. The study support that in the upright standing giraffe cerebral blood flow



                                                  75
                                              Abstracts

is governed by arterial pressure without support of a siphon and that blood accumulates in the vein
when the head is lowered.

S1703
Orthostatic challenges of the snake’s cardiovascular system
WANG T, ANDRADE D, ABE A, HICKS JW
University of Aarhus, Denmark; Universidade Estadual Paulista, Rio Claro, Brazil; University of
California, Irvine, USA

All 2700 species of snakes lack legs and have long elongate bodies. Some species may reach a
length of many meters and this morphology renders their cardiovascular system of snakes very prone
to the influence of gravity. Such influence may be aggravated by the low blood pressure that are
characteristic of ectothermic vertebrates and most snakes (cf. Wang et al., 2003). A long held
hypothesis proposes that the heart of arboreal snakes is positioned closer to the head than terrestrial
species, which was speculated to be an adaptation that secures the cerebral perfusion when arboreal
snakes lift their head (Lillywhite, 1987). A recent phylogenetic analysis of more than 150 species of
snakes belonging to various families, which takes the evolutionary history of the various species into
account, however, shows that the heart of arboreal snakes is placed more posteriorly than terrestrial
species (Gartner et al, in review). We have investigated the effects of head position on blood
pressures, cardiac output and blood flow in the carotid artery of anaesthetised rattlesnakes and show
that blood flow to the brain is severely compromised when the head is lifted above heart level. We
also have indications that blood pools in the posterior part of the body when the rear end of the snake
is lowered below heart level, which caused a decrease in cardiac filling and stroke volume.

Gartner, G.E.A., J.W. Hicks, P.R. Manzani, D.V. Andrade, A.S. Abe, T. Wang, S.M. Secor and
T.Garland Jr. Phylogeny, Ecology, and Heart Position in Snakes. Physiological and Biochemical
Zoology (in review).
Lillywhite, H.B. (1987). Circulatory adaptations of snakes to gravity. American Zoologist 27:81-95.
Wang, T., J. Altimiras, W. Klein and M. Axelsson (2003). Ventricular haemodynamics in Python
molurus: separation of pulmonary and systemic pressures. Journal of Experimental Biology 206:
4241-4245.

S1704
Abstract missing

S1705
The supine position is the normal position for humans.
SECHER NH
Department of Anaesthesia, Rigshospitalet, University of Copenhagen, Denmark.

When humans stand or are seated, they may faint. In these positions approximately 80 % of blood
volume is below the level of the heart. Veno- and vasoconstriction together with the veno-arterial
reflex cannot hinder critical accumulation of blood in dependent parts of the body. Venous return
needs to be supported by the muscle pump and yet, in an upright or seated position cardiac output is
preload dependent even during exercise. The “plateau” of the Starling curve for the heart is reached
during supine rest. Volume regulating hormones respond to the central blood volume and the

                                                  76
                                              Abstracts

reduced central blood volume of upright humans explains why the total blood volume is larger (7 %
body weight) than for most animals (5 %). In the upright position, thirst and reduced diuresis
enhance plasma volume. Together these observations indicate that normovolaemia may be defined
for supine humans as the blood volume that does not limit cardiac output. This consideration is
supported by the finding that surgical patients treated according maintaining a preload to the heart
that does not limit cardiac output in so-called individualized goal-directed volume therapy
demonstrate a reduced frequency of complications and even a reduced hospital stay. An explanation
for that finding is that regional flow, including cerebral blood flow, seems to be restricted whenever
cardiac output is limited. Accordingly, in regulation of regional flow, filling of the heart needs also
to be considered and in supine humans stroke volume of the heart independent of its preload or its
diastolic filling.




                                                  77
                                             Abstracts


                         Symposium 18: Neurosteroids in brain function

S1801
Neurosteroids: neurone specific endogenous modulators of the GABA-A receptor
BELELLI D, LAMBERT JJ, HERD MB, MITCHELL EA, GUNN BG, BROWN AR
University of Dundee, Neurosciences Institute, Div. of Pathology and Neuroscience, Ninewells
Hospital, Dundee, DD1 9SY, Scotland

It has been recognised over the past 20 years that that certain steroids synthesised de novo in the
brain (hence named neurosteroids) can produce immediate changes (within seconds) in neuronal
excitability, on a time scale that precludes a genomic locus of action. Identified molecular targets
underlying modulation of brain excitability, include both the inhibitory GABA-A and the excitatory
NMDA receptors. Of particular interest is the interaction of certain neurosteroids with the GABA-A
receptor, the major inhibitory receptor in the mammalian brain. During the last few years compelling
evidence has been presented demonstrating that neurosteroids act locally to selectively “fine tune”
neuronal inhibition. A range of molecular mechanisms including the subunit composition of the
receptor(s), phosphorylation mechanisms and local metabolism govern the non-genomic modulation
of synaptic and extra-synaptic GABA-A receptors by a range of endogenous and synthetic
neuroactive steroids and underpin their region and neurone-selectivity (Belelli & Lambert 2005;
Herd et al. 2007). Here, we will review the current understanding of the relative impact each of these
mechanisms has on neurosteroid action within specific neuronal circuits and the relevance of these
effects to the role of neurosteroids in health and disease. References: Belelli, D. & Lambert, J.J.
2005. Nature Rev. Neurosci. 6(7), 565-75. Herd, M.B., Belelli, D. & Lambert, J.J. 2007. Pharmacol.
Ther. 116(1), 20-34.

S1802
Neurosteroids in neuronal damage and repair
SCHUMACHER M, LABOMBARDA F, LIERE P, DE NICOLA AF. GUENNOUN R
UMR 788 Inserm and University Paris-Sud 11, Kermlin-Bicetre, France

The view that gonadal steroids are only reproductive hormones has completely changed over the past
years. A large number of experimental studies, most performed in rodents, have indeed demonstrated
the multiple actions of steroids throughout the nervous system. Thus, progesterone, a steroid
hormone well known for its role in the maintenance of pregnancy, has been shown to promote the
viability and regeneration of neurons and also to play a role in the formation of myelin sheaths,
which protect the axons and are required for the efficient conduction of electrical impulses along
nerve fibers. Pregnenolone, progesterone and its reduced metabolites are qualified as
"neurosteroids", because they can be synthesized by neurons and glial cells within the nervous
system. In response to traumatic brain injury or spinal cord lesion, the local formation of
pregnenolone and progesterone is increased, strongly suggesting that this response may be part of
the mechanisms by which nerve cells cope with neurodegeneration. However, the increase in
endogenous neurosteroids in response to a lesion is not sufficient for optimal neuroprotection or
myelin repair, as the administration of exogenous progesterone has marked protective and trophic
effects on neurons and oligodendrocytes. There is thus a potential for therapeutic interventions to
promote neuroprotection and neuroregeneration, either by stimulating the synthesis of endogenous
neurosteroids or by administering exogenous progestagens.


                                                 78
                                             Abstracts



S1803
Mechanisms of actions of neurosteroids in GABA-A receptor mouse model systems
LEPPÄ E, LINDEN AM, VEKOVISCHEVA OY, WULFF P, WISDEN W, KORPI ER
Institute of Biomedicine, Pharmacology, Biomedicum Helsinki, POB 63 (Haartmaninkatu 8), FI-
00014 University of Helsinki, Finland

Neurosteroids have non-genomic rapid actions on the brain via inhibitory -aminobutyric acid type
A (GABAA) receptors. These actions are potent and widespread. The actions of neurosteroids at
neuronal level depend on the ,  and subunits of the receptor. We have produced a mouse line
without GABAA receptor-mediated synaptic inhibition on cerebellar Purkinje cells by cell type-
specific ablation of the GABAA receptor 2 subunit, leaving only low conductance extrasynaptic -
GABAA receptors. These mice showed no obvious motor phenotype (Wulff et al., 2007) and their
GABAA receptor distributions as assessed by t-butylbicyclophosphoro-[35S]thionate autoradiography
were similar to wild-type littermates. However, the motor-impairing effect of the neurosteroid 5-
pregnan-3-ol-20-one in the rotarod test was dramatically increased in the mutant mice. By restoring
the 2 subunit in Purkinje neurons, the neurosteroid sensitivity could be diminished to control level.
The results suggest that neurosteroids are very efficacious potentiators of the remaining
extrasynaptic - GABAA receptors in this single neuronal population. References: Wulff P, Goetz
T, Leppä E, Linden AM, Renzi M, Swinny JD, Vekovischeva OY, Sieghart W, Somogyi P, Korpi
ER, Farrant M, Wisden W 2007. Nat Neurosci 10, 923-29.

S1804
Mechanisms for neurosteroid action on cognitive function from receptors to human disorders
STRÖMBERG J, LUNDGREN P, HAAGE D, TAUBE M, WANG M, BÄCKSTRÖM T
Umeå Neurosteroid Research Center, Umeå University, Dept. of Clin.Sci., Obstetrics &
Gynecology, 5B level 5, SE-901 85 Umeå, SWEDEN

GABA-A receptor (GABA-A-R) agonistic modulators e.g. benzodiazepines, barbiturates and
alcohol, used chronically give permanent memory and learning impairment, and increased risk for
dementia. The progesterone metabolite allopregnanolone (ALLO) is a GABA-A-R agonist (GABA-
steroid) and impairs learning in rats in the Morris Water Maze. In humans medroxy- progesterone
doubles the dementia frequency in 5 years. Chronic stress, burnout-syndrome and adrenal steroids
are linked to development of dementia, especially Alzheimer type. During stress a high number of
steroid hormones are produced. They are metabolized to 3-OH- 5-reduced steroids several being
modulators of the GABA-A-R function. Pregnane steroids with 3-hydroxy-configuration (3-
steroids) have been shown to reduce the ALLO enhanced GABA effect. The aim of our studies has
been to investigate the mechanism of the agonistic effect by ALLO and the antagonistic effect by
3-steroids. The 3- isomer of ALLO reduced the efficacy of ALLO in a non-competitive way.
Further, 3-steroids increased the desensitization rate of current response. By the [35S]TBPS-binding
assay it is possible to study the agonistic effect of ALLO but not the antagonistic effect of the 3-
isomer of ALLO. The mechanism of the 3-steroid is therefore suggested being desensitization
dependent in contrast to ALLO, which has been suggested to decrease the GABA unbinding rate.
The knowledge that diversity of endogenous steroids interact with the GABA-A-R function is of



                                                 79
                                             Abstracts

importance for further understanding of different sex and stress steroid related symptoms and
disorders.




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                                              Abstracts


                 Symposium 19: Testosterone and male reproductive physiology

S1901
Androgen signaling
JÄNNE OA
Biomedicum Helsinki, Institute of Biomedicine (Physiology), University of Helsinki, FI-00014
Helsinki, Finland

Androgens sculpture the male and female body in a number of ways, and androgen receptor (AR)
regulates these events in a tissue-specific fashion by activation or repressing specific genes and gene
networks. However, the regulatory mechanisms involved are not well understood. AR is a hormone-
dependent transcription factor that belongs to the superfamily of nuclear receptors. The
pharmacology of steroid hormones, such as androgens, is tripartite, in that it includes hormone-
dependent interactions with (i) the receptor, (ii) DNA sequences, and (iii) coregulatory proteins. For
many known nuclear receptors, including the steroid receptor subfamily (androgen, estrogen,
progesterone, glucocorticoid, and mineralocorticoid receptors), current evidence points to formation
of large, multi-component complexes in the nucleus, at the promoter and/or enhancer region of a
target gene, to elicit regulation of gene expression. This presentation deals with various aspects of
androgen signaling and highlights some novel aspects of AR function that have emerged from recent
genome-wide studies on AR binding and androgen signaling.

S1902
How much testosterone does the man need?
HUHTANIEMI I
Department of Reproductive Biology, Imperial College London, Hammersmith Campus, Du Cane
Road, London W12 0NN, UK

Androgens have two principal actions in the male, i.e. to maintain spermatogenesis in the testis and
to exert anabolic- androgenic effects in peripheral tissues. The levels of androgens in different
compartments of the body vary greatly, and also throughout life their levels undergo dramatic
changes. Therefore, it is relevant to ask what the necessary levels of testosterone are to maintain its
diverse functions. The concentration of intratesticular testosterone is about 100-fold higher than in
the periphery. It has been proposed that such a high intratesticular testosterone level is needed to
maintain spermatogenesis, but a simpler explanation is that the level is high only because the testis is
the site of synthesis of this hormone for the needs of the whole body. Another controversy surrounds
the importance of the declining androgen levels in ageing men. Do they mean that hypogonadism
upon ageing is a common phenomenon, in analogy with female menopause, or do the lower
androgen levels in ageing men maintain their peripheral anabolic-androgenic status? The purpose of
this lecture is to discuss the varying needs of androgens in the testis and periphery at different times
of the man‟s life span, based on recent experimental and clinical data.

S1903
Developmental effects of anti-androgens on reproductive system
TOPPARI J
Departments of Physiology and Pediatrics, University of Turku, FI-20520 Turku, Finland



                                                  81
                                               Abstracts

Male sexual differentiation is critically dependent on androgen action. Anti-androgens cause
disorders of male sexual development that can appear as maldevelopment of the penis (hypospadias,
micropenis) and maldescent of the testes (cryptorchidism). In females the anti- androgens have no
effects. There is a critical time-window in fetal development when adverse outcomes can rise as a
result of anti-androgen action. In addition to direct androgen action, several other mediators play a
role, such as desert hedgehog signaling and insulin-like factor 3 action. Several environmental anti-
androgens show dose-additive effects. In human studies, cryptorchid boys tend to be exposed to
higher levels of anti-androgens than healthy boys. It remains to be shown, whether these associations
occur by chance or whether there is a true cause-effect relationship.

S1904
Androgen receptor polymorphism and male reproductive health
GIWERCMAN A
Reproductive Medicine Centre, Malmö University Hospital, Lund University, Malmö, Sweden

The biological actions of androgens are mediated through the androgen receptor, which is a member
of the nuclear receptor family and is encoded by the androgen receptor gene (AR), located on the
long arm of the X chromosome. Exon1 of the AR has two polymorphic regions comprised by
repetitive sequence, the CAG and the GGN repeats, encoding for (poly)glutamine and (poly)glycin,
respectively. In vivo as well as in vitro studies have indicated an association between the CAG
repeat number and the transactivating activity of the AR. On the other hand, little was known about
the functional impact of the variation in the GGN number.
Based on in vivo and in vitro data we have provided evidence that the most common GGN alleles,
GGN=23 is associated with the most optimal function of the receptor. Subjects with higher or lower
GGN number are at higher risk of some of following reproductive disorders: lower ejaculate volume,
infertility, cryptorchidism and/or hypospadias. CAG repeat number may not only regulate the
function of the androgen receptor but also modify the effect of the persistent organohalogen
pollutant on the receptor function thereby adding to the inter-individual variation in the susceptibility
to such chemicals. An evidence for such effect was found on an epidemiological level and also in
vitro One of the mechanisms behind this gene-environment interaction might be CAG repeat number
dependent effect of co-factors on the receptor function. Although the physiological role and
mechanisms behind the association between the AR polymorphisms and the receptor function is not
completely understood, the available data indicate that digging into these processes may clinically
and biologically important information regarding the regulation of male reproductive health.




                                                   82
                                              Abstracts


                                         Free oral session 01

F0101
Luminal glucose is a prerequisite for a stimulation of duodenal alkaline secretion by orexin-A
BENGTSSON MW, MÄKELÄ K, HERZIG K-H, FLEMSTRÖM G
Department of Neuroscience (Physiology), Box 572, SE-751 24 Uppsala, Sweden

Background & aim: The orexogenic peptide orexin-A is expressed in endocrine cells in duodenal
mucosa and stimulates duodenal alkaline secretion. However, stimulation occurs only in animals
with continuous access to food; overnight fasting thus abolishes the response to orexin-A. Our aim
was to identify luminal factors regulating this feeding-status dependency. Methods: Different
isocalometric nutrients (glucose, lactalbumin and corn oil) were given to overnight fasted Lewis x
Dark Agouti rats by enteral feeding. Animals were then anesthetized and proximal duodenum
cannulated in situ. Titration of duodenal alkaline secretion was started 4 hours after gavage and
orexin-A was administered to the duodenum by close intra-arterial infusion. Total RNA was
extracted from enterocytes isolated from continuously fed or overnight (16 hours) fasted animals and
expression levels of orexin receptors (OX1R and OX2R) measured by quantitative real-time PCR.
Results: Pre-administration of glucose but neither of protein nor of fat induced ability of duodenum
to respond to orexin-A (240 pmoles/kg × h) with a rise (p<0.01) in bicarbonate secretion.
Administration of glucose directly into the luminal perfusate shortly before start of titration was also
tested but did, in contrast to preceding gavage of glucose, not induce any response to orexin-A.
Feeding significantly increased (p<0.001) enterocyte expression of both orexin receptors.
Conclusion: Orexin-A induces duodenal alkaline secretion only in animals pretreated with glucose.
The required time interval between glucose exposure and sensitivity to orexin indicates need of gene
transcription. It would thus seem likely that gene transcription of duodenal orexin receptors is
regulated by a glucose sensitive pathway.

F0102
Apelin induced stimulation of duodenal bicarbonate secretion
FLEMSTRÖM G, BENGTSSON MW, JEDSTEDT G, MÄKELÄ K, HERZIG K-H
Physiology/Neuroscience Uppsala University BMC, POB 572 SE-751 23 Uppsala Sweden

Apelin is the endogenous ligand of the G protein-coupled receptor APJ and apelin peptide as well as
APJ mRNA are expressed in several tissues. Proposed actions include involvement in the control of
appetite and body metabolism. The orexogenic hormone orexin-A and the incretin GIP stimulate the
bicarbonate secretion by the duodenal mucosa, but stimulation occurs only in fed animals. Our aim
was to study effects of apelin on the duodenal secretion in fed and overnight food deprived animals.
Methods: Lewis x Dark Agouti rats had free access to water and, unless fasted overnight, free access
to food. Animals were anesthetized and a segment of proximal duodenum with intact blood supply
cannulated in situ. Mucosal bicarbonate secretion was titrated (pH stat) and apelin-13 was
administered to the duodenum by close intra-arterial infusion. Total RNA was extracted from
mucosal specimens, reverse transcripted to cDNA and expression of APJ receptor measured by
quantitative real-time PCR. Results: Apelin caused a 30–40 % rise in secretion at the lowest dose
infused (6 pmol/kg × h). A 10-fold higher dose (60 pmol/kg × h) caused an additional slight increase
in secretion but a 100-folder higher dose had no further effect. Stimulation occurred only in fed
animals. No stimulation was thus observed in overnight fasted animals, even with the highest dose of


                                                  83
                                              Abstracts

apelin tested (600 pmol/kg × h). Pretreatment with atropine did not affect the secretory response to
apelin in fed animals. Overnight fasting caused a 8-fold decrease in the expression of APJ receptor
mRNA. Discussion. Very low doses of apelin stimulate the bicarbonate secretion by the duodenal
mucosa. Stimulation does not involve muscarinergic pathways and is, as found with orexin-A and
GIP, markedly dependent on feeding status.

F0103
LPS and PGN stimulates mucus secretion in colon
PETERSSON J, LUNDBERG J, HOLM L, PHILLIPSON M
Dept of Medical Cell Biology, Uppsala University, Uppsala, Sweden

Introduction: The colon, as the rest of the GI tract, is covered with a firmly adherent mucus layer
which serves as a physical barrier. The mucus layer is important in the colon for preventing colonic
bacteria from invading the colonic mucosa and cause inflammation. The regulation of the mucus
secretion is poorly understood. In this study, we wanted to investigate if the mucus secretion might
be regulated by luminal bacterial products. Methods: Germ Free NMRI mice and conventional
NMRI mice were used. The descending colon of anaesthetized (Isoflurane ®) mice was exteriorized
and the mucosal surface visualized. The firmly adherent mucus thickness was measured with
micropipettes after which the loosely adherent layer was removed by suction. The accumulation of
the mucus was measured for 45 min, and the firmly adherent mucus layer was then measured again.
In another set of animals PGN or LPS was applied luminally during the 45 min period. Results: The
Germ Free animals had hardly any firmly adherent mucus in the descending colon (8±0 µm), while
the convention animals had a 34±1 µm thick adherent mucus layer. When the colonic mucosa of the
germ free animals was exposed to LPS or PGN luminally, the thickness of the adherent mucus layer
increased to 29±3 µm and 35±1 µm respectively. This effect could not be observed in the
conventional animals, which when treated with LPS had an adherent mucus layer as same as in the
control situation (33±2 µm). Conclusion: When treating the germ free mice with LPS and PGN
luminally, the firmly adherent mucus layer increases to the levels that we normally can observe in
the conventional mice. Since LPS is a TLR2 ligand and PGN is a TLR4 ligand, the mucus secretion
could be regulated by bacterial endotoxins or plasma membrane via the TLR2 and TLR4 receptor
pathways.

F0104
Genetic ablation of the K+ channels Kir 4.1 and KCNQ1 has opposing effects on acid secretory
rates in gastric mucosa of weanling mice
SONG PH, GROOS S, RIEDERER B, KRABBENHOFT A, ENGELHARDT R, MANNS MP,
SMOLKA AJ, NEUSCH C, SEIDLER U
Dept. of Gastroenterology, Hepatology, and Endocrinology, Hannover Medical School, 30625
Hannover, Germany
Background: K+ recycling over the apical membrane is essential for gastric acid secretion, and both
Kir 4.1 and KCNQ1 K+ channels have been recently found to traffic to the parietal cell apical
membrane during acid stimulation. Aims: To understand the physiological role of these channels by
studying the acid secretory capacity and parietal cell morphology of very young Kir 4.1- and
KNCQ1- deficient, heterozygotic and wildtype (WT) mice. Methods and Results: At very young age
(7-9 days), KCNQ1 -/- gastric mucosa in Ussing- chambers was able to secrete acid, but basal and
forskolin-stimulated acid secretory rates were strongly reduced in KCNQ1-/- compared with +/+ and

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                                             Abstracts

+/- mice, whereas the Isc response, indicative of electrogenic anion secretion, was significantly
increased. Electron micrographs displayed similar PC cell numbers in +/+ and -/- stomach, but
hyperproduction of tubulovesicles and secretory membranes, as well as membrane filled
autophagosomes in KCNQ1-/- parietal cells. Surprisingly, Kir 4.1 -/- mucosa secreted significantly
more (24 %), and, more importantly, much faster (238 % of control increase after 10 min) acid, with
a significant overshoot upon forskolin-stimulation compared to +/+ and +/- mucosa, despite the fact
that the Kir 4.1 -/- mice were smaller and the gastric mucosa thinner than the WT mice. Conclusions:
KCNQ1 channels are crucial for gastric acid secretion at a very young age by regulating K+
recycling and maintaining the viability of tubulovesicles and canaliculi in parietal cells. Kir 4.1
channels have a distinctly different function, seemingly acting as a "brake" on initial proton pumping
and thus ATP consumption.

F0105
Angiotensin II regulates the mRNA expression of enzymes involved in arginine metabolism in
isolated renal resistance vessels through the AT1 receptor
HULTSTRÖM M, IVERSEN BM
Renal research group, Dept. of Medicine, Haukeland University Hospital, 5021 Bergen, Norway

We previously found equally increased expression of L-arginine metabolising enzymes in both
kidneys of rats with induced renal hypertension implied a direct role of Angiotensin II (AngII) in
their regulation. The aim of the present study was to investigate the expression of these genes in
AngII infused rats. 40 male wistar rats were implanted with Alzet micropumps and infused with
either 80 ng/min AngII (n=20) or saline (n=20) for 14 days. Ten animals from each group received
the AngII AT1 receptor blocker Losartan (60 mg/kg/day) in the drinking water. Blood pressure was
monitored using the tail cuff method. After 14 days the rats were sacrificed and preglomerular
vessels were isolated. mRNA expression was investigated using RTPCR with 18S RNA as internal
control. Mean arterial pressure (MAP) was increased by AngII infusion from 102±6 mmHg to 147±9
mmHg (P<0.05). Losartan decreased MAP in Ang II treated rats (94±6 mmHg, P<0.05) but had no
effect in controls (101±4 mmHg). The expressions of eNOS (0.88±0.26 vs. 1.1±0.34), DDAH-1
(1.47±0.41 vs. 1.58±0.45) and Arginase-2 (1.46±0.62 vs. 1.8±0.78) were not changed by AngII
infusion and were not affected by Losartan treatment. The expression of CAT-1 (0.38±0.07 to
0.73±0.12, P<0.05), CAT-2 (1.14±0.29 to 2.74±0.48, P<0.05), DDAH-2 (1.09±0.27 to 2.3±0.46,
P<0.05) and Arginase-1 (1.08±0.17 to 1.82±0.22, P<0.05) were significantly increased in AngII
infused rats and the increase was abolished by Losartan treatment. In conclusion, the mRNA
expressions of CAT-1, CAT-2, DDAH-2 and Arginase-1 in renal resistance vessels are regulated by
AngII stimulation of the AT1 receptor. This may be important for nitric oxide synthesis and the
regulation of renal blood flow.

F0106
Intact angiotensin II signalling in the postnatal period is necessary for normal development of
the renal microcirculation
MADSEN K, MARCUSSEN N, PEDERSEN M, SKØTT O, JENSEN BL
Department of Physiology and Pharmacology, Institute of Medical Biology, University of Southern
Denmark




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                                             Abstracts

Interruption of ANG II signalling in the postnatal period leads to pathological changes in the kidneys
characterized by impaired nephron formation, shortening and wall thickening of the preglomerular
arteries and papillary atrophy. The mechanism for this effect is not clear. We hypothesized that ANG
II is crucial for maintaining capillary angiogenesis during postnatal development. Rat pups were
treated with subcutaneous injections of AT1 receptor antagonist (candesartan 1 mg/kg×day) or
vehicle from postnatal day 1 (P1) to P13. On P14, pups were sacrificed and kidneys were analyzed
for vascular growth factors and postglomerular microvessel quantity. Quantitative PCR analysis of
cortex and medulla showed a significantly decreased expression of VEGF, angiopoietin-1 and -2 and
Tie-2. Unbiased stereological measurements showed a significant reduction in total capillary length
in cortex (288±19.6 m versus 211±17.8 m) and medulla (235±15.8 m versus 133±5.9 m). Vasa recta
bundle assembly was impaired by candesartan in outer medulla and microdissected bundles
exhibited fewer mitoses as judged by PCNA abundance. Renal blood flow (RBF) and GFR was
estimated in candesartan- treated pups (P1-P13) at P30 by Magnetic Resonance Imaging. RBF was
reduced significantly (~20 %) in the candesartan-treated group. GFR was unchanged. We conclude
that ANG II signalling is necessary for postnatal development of postglomerular capillaries and vasa
recta bundles and inhibition of AT1 receptors in the postnatal period leads to decreased RBF in early
adulthood.




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                                             Abstracts


                                        Free oral session 02

F0201
Optical mapping of the rabbit sinoatrial node under cholinergic and adrenergic influence
ABRAMOCHKIN DV, KUZMIN VS, SUKHOVA GS, ROSENSHTRAUKH LV
Biological faculty of MSU, Leninskiye gori, 1, 12, Moscow, Russia

The shift of the leading pacemaker site within the sinoatrial node (SAN) may act as a mechanism of
the sinus rhythm (SR) modulation. However, conventional methods of myocardium mapping do not
allow to observe beat-to-beat changes in the SAN activation sequence. Therefore, we applied high-
resolution optical mapping technique to register beat-to-beat changes in the SAN activation pattern
during cholinergic and adrenergic influence. Acetylcholine (10 μM) or stimulation of the intramural
parasympathetic nerves caused slowing of the SR as well as pacemaker shift (PS) and formation of
inexcitable region in the central part of the SAN. The latter effect may be one of the reasons of the
PS. The slowing of the SR, which exceeded 12.8±3.1 % of the control rate of the SR, was always
accompanied with PS. Isoproterenol (10 nM–1 μM) or stimulation of sympathetic postganglionic
nerves also evoked the PS, but without formation of the inexcitable zone. The acceleration of the SR
by more than 10.5±1.3 % of the control rate of the rhythm, always coincided with PS. Thus, either
cholinergic or adrenergic factors cause PS in the rabbit SAN. Modest changes of the SR does not
coincide with PS, but greater changes are always accompanied by the shift and may be due to it,
according to one of the hypothesis. Formation of inexcitable zone at the site of the basic leading
pacemaker location seems to be one of the mechanisms of the PS, but not the only.

F0202
Mitochondrial ROS triggered by β-adrenergic stress contributes to cardiac inotropy in wild-
type but not ob/ob mice
ANDERSSON DC, FAUCONNIER J, KATZ A, WESTERBLAD H
Karolinska Institutet, Dept. of physiology and pharmacology, von Eulers Väg 8, 17177 Stockholm,
Sweden

Objective: We studied if adrenergic stimulation induced production of mitochondrial reactive
oxygen species (ROS) in wild type (WT) and obese ob/ob mouse cardiac cells. Also, we studied if
ROS affected cardiac Ca2+ transients in adrenergic stimulation. Methods: Freshly isolated ob/ob and
WT cardiac cells were paced at 1Hz, cytoplasmic Ca2+ transients ([Ca2+]i) and mitochondrial
superoxide were measured in a confocal microscope using fluorescent dyes Fluo-3 and MitoSOX
Red, respectively. As a marker of oxidative stress, malondialdehyde (MDA) modification of proteins
was detected with western blotting. 100nM isoproterenol (ISO) was used as beta-adrenergic agonist.
Results: ISO increased ROS production in WT cardiac cells. Moreover, WT hearts perfused with
ISO for 30 min showed a doubling in MDA relative control. In WT hearts, the amplifying effect of
ISO on [Ca2+]i amplitude was diminished in presence of the antioxidant NAC (control: 4.40.32, ISO:
6.60.37, ISO+NAC: 5.20.33); a reduced effect on cell shortening was also seen (increased from
9.81.1 % to 17.91.2 % with ISO and 13.50.9 % with ISO+NAC). In contrast, ob/ob cells did not
significantly increase ROS production when exposed to ISO. Control experiments indicated a lower
rate of mitochondrial ROS production in ob/ob than WT hearts. The effect of ISO on [Ca2+]i
amplitude in ob/ob hearts was not significantly affected by NAC (control: 3.50.22, ISO: 6.00.39,
ISO+NAC: 5.60.18). Conclusion: ROS production in heart contributes to the inotropic effect of beta-


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                                               Abstracts

adrenergic agonists by increasing the amplitude of [Ca2+]i transients. In contrast to general belief,
our data suggest that obesity is associated with decreased ROS production in cardiac muscle.

F0203
Postnatal maturation of arterial smooth muscle (ASM): the impact of sympathetic
innervations
TARASOVA OS 1,2, PUZDROVA VA 1, GAINULLINA DK 1, KUDRYASHOVA TV1,
VOROTNIKOV AV 1, SCHUBERT R 3, AALKJAER C 4, NILSSON H 1
1
  M.V. Lomonosov Moscow State University, Leninskie Gogy 1/12, 119991, Moscow, Russia, 2 SRC
RF - Institute for Biomedical Problems RAS, Khoroshevskoe shosse 76A, 123007, Moscow, Russia, 3
University of Rostock, Germany

Rapid ASM contraction and relaxation is essential for dynamic sympathetic control of peripheral
resistance. Whereas contraction of ASM is triggered by an increase in free cytosolic Ca2+ ([Ca2+]i ),
the resultant forces may vary depending on Ca2+-sensitization of the contractile machinery. This
study tested the hypothesis that balance of Ca2+-dependent and Ca2+-independent mechanisms of
contraction is controlled by sympathetic innervation and changes during its development. We
demonstrate that establishment of sympathetic control of rat saphenous artery over the first month of
postnatal development is accompanied by Ca2+-desensitization of ASM contraction, which is
reversed by surgical denervation of the artery. The ASM from the adult rats, but not from their
denervated mates, required higher [Ca2+ ]i to generate isometric forces comparable to those produced
by ASM of the 1-2-week-old rats in response to the α- adrenergic or thromboxane receptor
activation. The progress of sympathetic innervation was associated with an increased expression of
myosin light chain kinase, myosin light chain phosphatase targeting subunit and h-caldesmon, i.e.
the proteins that mediate Ca2+-dependent regulation. In contrast, expression of Rho-kinase, ERK and
p38 MAP-kinases, that are involved in Ca2+-sensitization, was gradually diminished. The opposite
changes in the expression profiles of these proteins, associated with increased Ca2+ -sensitivity of
contraction, were observed upon denervation of ASM in the adult rats. Altogether, these data suggest
that sympathetic nerves exert trophic function in establishment of rapid contractile phenotype of
ASM. Supported by RFBR (grant 07-04- 01527) and cooperation program Uni-Rostock - MSU.

F0204
The role of Rho-kinase in regulation of Ca2+-sensitivity of saphenous artery contraction in
newborn and adults rats
MOCHALOV SV, SCHUBERT R*, TARASOVA OS
M.V. Lomonosov Moscow State University, Leninskie Gory 1/12, 119991, Moscow, Russian
Federation, *University of Rostock, PSF 100888, 18055, Rostock, Germany

The aim of this study was to explore the role of different contractile mechanisms of arterial smooth
muscle, with special emphasis on those regulating Ca2+-sensitivity, during postnatal development.
Rat saphenous arteries from newborn (NB: 5-10 day old) and adult (AD: 2.5-3.5 month old) animals
were studied using wire-myography and FURA-2 fluorimetry. In vessels from AD, methoxamine
(MX, 10 µM) and KCl (42 mM, in the presence of 5 µM guanethidine to block transmitter release
from periarterial nerves) increase tension and [Ca2+] i; the increase in tension was proportionally
larger than the increase in [Ca2+]i reflecting an augmentation of Ca2+-sensitivity. The Rho-kinase-
inhibitor Y27632 at 3 µM reduced the MX- and KCl-induced increase in tension and [Ca2+]i

                                                   88
                                              Abstracts

substantially; the effect on tension was proportionally larger than the effect on [Ca2+]i indicating an
additional, albeit not very prominent effect on Ca2+-sensitivity. In vessels from NB, MX (10 µM)
and KCl (42 mM) increase tension but do not change [Ca2+]i reflecting a considerable augmentation
of Ca2+- sensitivity. The Rho-kinase-inhibitor Y27632 reduced the MX- and KCl-induced increase in
tension without affecting [Ca2+] i indicating an effect on Ca2+-sensitivity. Thus, the results of this
study show that MX- and KCl-induced contractions of saphenous arteries are mediated by partly
Rho-kinase dependent changes of [Ca2+]i and Ca2+-sensitivity in adult animals and by almost
completely Rho-kinase dependent changes of Ca-sensitivity in newborn animals. This work was
supported by RFBR (grant 07-04- 01527) and by the program of cooperation between Uni-Rostock
and MSU.




                                                  89
                                              Abstracts


                                        Free oral session 03

F0301
Purinergic P2X and P2Y receptors – Opponents in neck muscle nociceptive processing
ELLRICH J, REITZ M, MAKOWSKA A, RISTIC D
Medical Physiology Group, Center for Sensory-Motor Interaction, Department of Health Science
and Technology, Medical Faculty, Aalborg University, Fredrik Bajers Vej 7D2, DK-9220 Aalborg,
Denmark

In muscle nociception, purinergic mechanisms play an important role. Studies on muscle pain
typically apply ,-meATP as noxious agent due to its narrow receptor profile (P2X3, P2X2/3) and
sustained stability in tissue. In contrast, native ATP is quickly degraded to metabolites and interacts
with excitatory P2X and inhibitory P2Y receptors. The experimental pharmacological study
compares effects of both molecules in myofascial nociception in a model of neck muscle pain.
Noxious stimulation of semispinal neck muscles was performed by intramuscular (i.m., 20 µl)
bilateral injection of ,- meATP (100 nM, 1µM) or native ATP (100 nM, 1 µM, 7.6 mM) in
anesthetized mice (n=65). The impact of neck muscle noxious input on brainstem sensory processing
was tested by the jaw-opening reflex elicited via electrical tongue stimulation. The P2Y1 receptor
antagonist MRS2179 (1 µM, 20 µl) or the P2Y1 receptor agonist 2-MeSADP (1 µM, 20 µl) were
i.m. administered 20 min before ATP or two hours after ,-meATP application, respectively.
Injection of ,-meATP facilitated the reflex within two hours in a dose-dependent manner. In
contrast, native ATP injection evoked facilitation only with low dosage (100 nM). Preceding
blockade of P2Y1 receptors by MRS2179 revealed reflex facilitation even under high dosage of
native ATP. Ongoing facilitation after injection of ,-meATP was abolished by subsequent
activation of P2Y1 receptors via 2- MeSADP. P2Y1 blockade reveals excitatory ATP effects on P2X
receptors. P2Y1 activation counteracts P2X3 excitation by ,-meATP. Results demonstrate
opposing excitatory P2X3 and inhibitory P2Y1 effects of ATP in neck muscle nociceptive
processing in mice. These mechanisms may be involved in the pathophysiology of neck muscle pain
in man.

F0302
Spatial organization of nociceptive long-term depression in man
JUNG K, ROTTMANN S, SIMONSEN LL, JOHANNESEN MD, KUNWALD MR, NIELSEN TB,
STEHR AM, ELMOSE SF, ELLRICH J
Medical Physiology Group, Center for Sensory-Motor Interaction, Department of Health Science
and Technology, Medical Faculty, Aalborg University, Aalborg, Denmark

Electrical low-frequency stimulation (LFS) of nociceptive afferents evokes long-term depression
(LTD) of pain. In-vitro studies suggest a sole homosynaptic effect on the conditioned pathway. The
spatial organization of LTD in man is still unclear. Thus, the present study addresses the hypothesis
that noxious LFS evokes homotopic LTD in man. In 36 healthy volunteers 86 psychophysical
experiments were performed. Painful electrical test stimulation and conditioning LFS (1 Hz, 1200
pulses, 4-fold pain threshold) were applied by a concentric electrode. Electrical stimulation was
performed on both dorsal hands (A), on right hand dorsum (B), or on low-back (C). Volunteers rated
electrically-evoked pain perception according to a verbal rating scale (0 to 100). (A) Test stimulation
was alternately applied to radial sides of right and left hand dorsum. After LFS on right hand dorsum

                                                  90
                                              Abstracts

pain rating significantly decreased (-26.8 %, p<0.001) exclusively on homotopic right hand. (B) Test
stimulation was alternately applied to radial and ulnar sides of right hand dorsum. After radial site
LFS pain rating significantly attenuated (- 44.1 %, p<0.001) exclusively within the same area. (C)
Test stimulation was performed within left dermatome T12 on low-back. LFS was applied either
within the same receptive field as determined by two-point-discrimination (T12) or remote in
dermatome T8. After LFS within dermatome T12 pain rating decreased (-18.8 %, p<0.05) only in
T12 but not in T8. Results indicate homotopic organization of LTD in healthy volunteers. Due to
large receptive fields in low-back area, even heterotopic LFS within the same receptive field evokes
LTD. Thus, data reveal homosynaptic spatial organization of somatosensory LTD in man.

F0303
ATP and nitric oxide (NO) interfere in microglial responses to spinal cord injury in vivo in
mice
SCHOMBURG ED*,DIBAJ P**, STEFFENS H*, NADRIGNY F, NEUSCH C**, KIRCHHOFF F
Max-Planck-Institute of Experimental Medicine and *Inst. of Physiology and **Dept. of Neurology,
University of Göttingen, D-37073 Göttingen, Germany

In the mouse cortex, microglial cells respond vividly to acute injuries of the grey matter in an ATP-
dependent manner. We now used 2-photon laser-scanning microscopy in mice with EGFP-labelled
microglia (MG) to study the interaction of purinergic and nitric oxide (NO) signalling systems in the
spinal white matter. In anaesthetized mice a laminectomy was performed at segment L4. Then MG
responses to acute laser induced micro-injuries were recorded at high temporal and spatial
resolution. Both, ATP and the volatile transmitter NO, tested as the NO-donor SPNO, attracted MG
processes when locally applied in the spinal cord by a micropipette. However, both compounds
could also act as suppressors of the MG response when the spinal cord below the laminectomy was
uniformly incubated with the drugs. To analyse the interference between purinergic and NO-
pathways, we combined the application of activating and blocking compounds for the two pathways.
MG attraction induced by local intraspinal ATP-application was not reduced when the spinal cord
was superfused with the guanylate cyclase blocker ODQ or SPNO. In contrast, the MG response to
local SPNO application was reduced when ambient ATP was removed by the ATPase apyrase; but,
the MG response to local SPNO-application was increased when the spinal cord was superfused with
ATP. We conclude that ATP exerts its effect on MG independently of the NO-pathway, while
components of the purinergic signal cascade modulate the MG attraction by NO.

F0304
Pancreatic islet revascularisation in the intramuscular transplantation site studied in vivo
CHRISTOFFERSSON G, HENRIKSNÄS J, CARLSSON PO, PHILLIPSON M
Department of Medical Cell Biology, Division of Integrative Physiology, Uppsala University, Box
571, 751 23 Uppsala, Sweden

The pursuit of new implantation sites for transplantation of islets of Langerhans requires new
experimental models. Since today‟s clinical islet transplantations to the liver show low success rates
in part due to low revascularisation, we have developed a new in vivo-method for studying the
revascularisation of islets transplanted to muscle. Mouse islets of Langerhans were isolated and
syngeneically transplanted to the cremaster muscle or liver of mice. At different time-points post-
transplantation the mice were anesthetized and the muscle was prepared for intravital and confocal

                                                  91
                                              Abstracts

microscopy. Revascularisation and microcirculation of the islets were examined and the graft-
bearing organs were also retrieved and prepared for immunohistochemistry. Islets transplanted to the
cremaster muscle had improved revascularisation compared to islets transplanted intraportally to the
liver. Grafts in muscle had functional intra-islet vessels three days post-transplantation, whereas the
newly formed vessels to the grafts in the liver instead surrounded the islet. Intravital microscopy of
islets in the cremaster muscle revealed that leukocyte-endothelial cell interactions were increased in
venules draining the transplanted islets compared to venules draining only the muscle. Emigrated
leukocytes were also seen inside the grafts and in the adjacent tissue. In conclusion, the muscular site
of engraftment was beneficial in terms of intra-islet revascularisation compared to the liver site. This
new model offers a way to investigate pancreatic islet engraftment after transplantation to muscle in
vivo. Vessel growth and -density are easily quantified and the model enables studies of the complex
courses of events in graft angiogenesis, including contributions from different immune cell subsets.




                                                  92
                                             Abstracts


                                        Free oral session 04

F0401
The effect of adenosine, hypoxia and exercise on local skeletal muscle blood flow in humans
HEINONEN I, KEMPPAINEN J, KASKINORO K, PELTONEN J, LINDROOS M, BORRA R,
HELLSTEN Y, BOUSHEL R, KALLIOKOSKI K
Turku PET Centre, PO Box 52, FIN-20521, Turku, Finland

Introduction - The effect of maximal pharmacological stimulus and physiological systemic hypoxia
on blood flow locally within skeletal muscle is elucidated in the present study for the first time in
humans. Methods - Seven healthy young men were studied with positron emission tomography.
Skeletal muscle blood flow and oxygen consumption as well as several other parameters were
measured at rest, during femoral arterial adenosine infusion, in systemic hypoxia (14 % O2) at rest,
and during one-legged dynamic exercise in normoxia and hypoxia without and with adenosine
receptor blockade. Whole body VO2max bicycle test and leg MRI were also performed. Results -
Adenosine increased muscle blood flow almost 20-fold at rest and muscle oxygenation was virtually
entirely provided by blood flow since oxygen extraction was reduced close to zero. However, blood
flow distribution seemed not to change substantially despite large increases in flow. Systemic
hypoxia caused significant drop in arterial oxygen saturation from 98±1 % to 90±5 % at rest but
muscle blood flow remained unchanged while venous oxygen content was reduced as compared to
normoxia. There were also significant changes in different parameters due to exercise, but the uptake
of energy substrates changed surprisingly little despite substantial changes in blood flow and
metabolic conditions. Discussion - Adenosine has potential to increase muscle blood flow
extensively, but systemic hypoxia corresponding 3000 m above sea level appears to have little effect
on blood flow locally in skeletal muscle.

F0402
Cardiac output, common and deoxygenated hemoglobin in working muscle during incremental
bicycle test in subjects of various aerobic performance levels
POPOV D, MARCHENKO D, BOROVIK A
76A, Khoroshevskoe shosse, Moscow, Russia

Oxygen consumption of working muscles depends on oxygen delivery and oxidative capacity of
muscle. The aim of the study was to compare dynamics of cardiac output (CO), blood filling of
working muscle and muscle oxygen consumption evaluated by common hemoglobin (cHb) and
deoxygenated hemoglobin (HHb) accordingly during incremental test. 7 young men and 8 athletes
performed bicycle incremental ramp test till exhaustion. CO was determined up to maximal aerobic
power (MAP) by open-circuit method using gas mass-spectrometer. HHb and cHb concentrations
were continuously evaluated in m. vastus lateralis (VL) by near- infrared spectroscopy. HHb in
athletes increased linearly till exhaustion. Untrained subjects demonstrated slowing down of HHb
increase at submaximal aerobic power. The observed differences could be connected with low
oxidative capacity of muscle or with restricted oxygen delivery to the muscle. Indeed untrained
subjects demonstrated slowing down of cHb in VL at 50-80 % of MAP, while the athletes showed
an increase of cHb up to 90-100 % of MAP. Half of the subjects demonstrated slowing down of CO
increase at 90-100 % of MAP. The dynamics of CO was not related to dynamics of cHb in VL. That
means that oxygen delivery to working muscle at maximal aerobic power may be limited both by


                                                 93
                                              Abstracts

CO and/or by redistribution of blood from working muscles to other regions, for example, to
respiratory muscles. Thus comparison of CO dynamics with that for cHb and HHb in working
muscle allows to reveal the factors limiting individual aerobic performance. The work was supported
by RFBR grant #60-04- 49699a.

F0403
Effects of repeated bout exercise on inhibition of extracellular matrix degradation on rat
muscle
KOSKINEN S, HESSELINK M, AHTIKOSKI A, KOMULAINEN J, TAKALA T
Department of Sports Medicine, Oulu Deaconess Institute, Oulu, Finland, Department of Health
Sciences, University of Oulu, Finland

It has been shown that prior forced shortening and lengthening contractions may result to more
pronounced upregulation in type IV collagen degradation capacity after repeated bout of
contractions. The purpose of the present study was to investigate whether type of previous exercise
affects on the changes caused by lengthening contractions in type IV collagen degradation capacity
measured as expression of tissue inhibitors of metalloproteinases (TIMP). The left TA muscles of 12
week-old male Wistar rats were subsequently subjected to 240 forced shortening or lengthening
contractions. Tetanic contractions were induced by electrical stimulation of the common peroneal
nerve. Shortening or lengthening of TA muscles were performed by either plantar or dorsiflexion of
the foot at the ankle joint with rotational velocity of 500 °/s. 15d later half of the both groups were
subjected to second bout of exercise, forced lengthening contractions. The contralateral leg was used
as a control. Histological examination of TA showed severe skeletal muscle fiber injury 4d after
forced lengthening contractions and both types of repeated bout of exercise (shortening +
lengthening or lengthening + lengthening), but only slight histopathological changes after forced
shortening contractions. MMP-2 inhibitory activity of TIMP-2 elevated remarkable 4d after both
shortening and lengthening contractions. TIMP-2 was still elevated 15d and 30d after single bout of
exercise. Interestingly, the second bout of exercise (lengthening contractions) 4d after did not cause
as remarkable increase as the single bout exercises. Actually, MMP-2 inhibitory activity of TIMP-2
was in the same level as at 15d after single bout exercise. Changes in inhibitory activity of TIMP- 1
were more moderate than changes in TIMP-2. Together the results suggest that MMP degradation
capacity is higher after repeated bout exercise than after single bout exercise.

F0404
Ketone bodies inhibit insulin-mediated glucose transport in mouse skeletal muscle
KATZ A, YAMADA T, ZHANG S-J, WESTERBLAD H
Karolinska Institutet, Department of Physiology and Pharmacology, 171 77 Stockholm, Sweden

Ketone bodies (KBs) serve as alternative substrates when glucose availability is compromised (e.g.
starvation). It is unclear whether KBs directly contribute to insulin resistance. We investigated the
effects of D,L- beta-hydroxybutyrate (BOH, the major KB in vivo) on glucose transport in isolated
mouse soleus (oxidative) and extensor digitorum longus (EDL, glycolytic) muscle. BOH did not
alter glucose transport (2-deoxyglucose uptake) in a noteworthy fashion in EDL muscle under any
condition studied, but inhibited insulin- mediated transport in soleus in a time and concentration-
dependent manner. Under optimal conditions (19.5 h exposure to 5 mM BOH), insulin (20 mU/ml)-
mediated glucose transport was almost fully inhibited (control=6.1±0.4 µmol/ml intracellular

                                                  94
                                            Abstracts

water/30 min; BOH=2.8±0.2; P<0.001). The inhibitory effect was reproduced with D- but not with
L-BOH. BOH did not significantly affect hypoxia- or AICAR (activates AMP-dependent protein
kinase)- mediated glucose transport. The BOH effect did not require the presence or utilization of
glucose since it was also seen when glucose in the medium was substituted with pyruvate. The BOH
effect was not reversed by an exogenous antioxidant and was not associated with increased
production of reactive oxygen/nitrogen species. BOH did not alter the levels of total tissue GLUT-4
protein, but blocked the insulin- mediated phosphorylation of PKB by almost 50 %. This inhibition
was not associated with the phosphorylation of PKC delta. These data demonstrate that BOH inhibits
insulin-mediated glucose transport in oxidative muscle by inhibiting insulin signaling. Thus KBs
may be potent diabetogenic agents in vivo.




                                                95
                                              Abstracts


                  Poster session 01 – Cardiovascular & pulmonary physiology

P01
Inflammation and electrical instability in chronic myocardial infarction
MOZOS I, HANCU M, CRISTESCU A
Department of Pathophysiology, T. Vladimirescu Street 14, Timisoara 300173, Romania

Inflammatory markers are considered in cardiovascular disease risk assessment and inflammation
has an integral role in all aspects of coronary syndromes. It is the aim of this study to find the
relationship between some inflammatory markers and electrical instability parameters in chronic
myocardial infarction (CMI) patients. 24 CMI patients underwent 12 lead ECG (QT dispersion as
the difference between the longest and shortest QT interval), signal averaged ECG (SAECG) (late
ventricular potentials: LVP) and 64 electrodes body surface mapping (BSM) (isointegral and
isopotential maps). We considered a patient having LVP (16 patients: 67%), if two of the following
criteria were positive: QRS duration (SA-QRS) >120 ms, LAS40 (the duration of the signal at the
end of the QRS complex with an amplitude below 40 μV) > 38 ms and RMS40 (the square root of
the last 40 ms of the signal) < 20 μV. White blood cell count (WBC: 9400±4100/mm3) and
erythrocyte sedimentation rate (ESR: 31±28 mm/h) were also assessed. There was a good correlation
between: ESR and SA- QRS (r=0.51), LAS40 (r=0.58), RMS40 (r=-0.52), QTdc (heart rate
corrected QT dispersion) (r=0.54), QRST isointegral maxima (r=0.52), QRS isointegral minima (r=-
0.60), isointegral Q40 minima (r=-0.76), QRS isopotential minima (r=- 0.60), ST isopotential
maxima (r=0.55). WBC correlated with QRST isointegral maxima (r=0.52), isointegral ST maxima
(r=0.55) and isointegral STT maxima (r=0.56). Inflammation increases sudden cardiac death risk in
CMI patients.

P02
Isolated factors identified in the effluent of preconditioned hearts offers cardioprotection to
donor hearts
BREIVIK L, HELGELAND E, STRIDH MH, HELGELAND G, MRDALJ J, SANDBERG M,
JONASSEN A
Department of Biomedicine, Medical Faculty, University of Bergen, Norway

One or several short episodes of non-lethal ischemic episodes administered to a heart prior (Ischemic
Preconditioning, IPC) or directly after (Ischemic Postconditioning, IschPost) a major ischemic
episode facilitates myocardic survival during the reperfusion phase. There is strong evidence that
this protection is due to a factor released from the heart during the conditioning phase. The perfusate
containing protects the heart when administered 10 minutes prior and after ischemia (25±3% and
30±3% vs. 55±3% for control, p < 0.05). The factor is so potent that when administered only in 3 x
30” bursts after ischemia it is enough to significantly lower infarct sizes (40±5% vs. control 56±4%,
p < 0.05. We have attempted to characterise and isolate this factor from effluate released during
preconditioning of isolated rat hearts in a Langendorff setup. We fractionated the proteins in the
effluent based on both size and hydrophobicity and the fractions were administered to unconditioned
rat hearts prior to a 30 min ischemic event and the infarct/risk ratio was calculated after 120 min
reperfusion. Hearts perfused with a hydrophobic fraction over 10 kDa had ischemic injuries at the
level of an IPC heart (25±6% vs. 22±3%, p < 0.05) and over two times lesser injuries than the hearts
that were perfused with a hydrophilic fraction and the hearts that were perfused solely with Krebs


                                                  96
                                              Abstracts

Heinsleitt Buffer (KHB) (25±3% vs. 55±3% and 50±12%, p < 0.05). We analysed the different
fractions by LC- MSMS and identified a total of 88 proteins released during preconditioning and of
several of them were only identified in fractions which were protective.

P03
Effects of purine compounds on bioelectrical activity of hibernating ground squirrel heart
KUZMIN VS, ABRAMOCHKIN DV
Moscow State University, Biological department, Vorob'evy gory 1, 12, Moscow, 119992 Russia

Hibernating animals over the course of evolution developed an adaptive winter slipping mechanism
which allows them to survive in the extreme condition of north. It is well known that adenosine and
purine nucleotides act as regulatory compounds in the cardiovascular system. Purine nucleotides‟
level is higher in the heart of hibernators in comparison with nonhibernators. Effects of purine
compounds on the heart of the hibernating animals are not sufficiently characterized. The aim of the
present study is to investigate effects of purine compounds as adenosine, AMP, ADP-ribose and
GMP (Sigma) on bioelectric activity of atrium and papillary muscle (PM) of the hibernating animal
(ground squirrel Citellus undulates) and the rat. Action potentials (AP) were obtained with use of
standard microelectrode technique. AP duration was estimated at the level of 50 % (APD50) and 90
% of repolarization (APD90) in the control experiments and in the presents of drugs (1×10-5M).
Purine compounds decreased APD50 and APD90 both in PM and atrium of hibernating ground
squirrel. In ground squirrel PM adenosine, AMP, ADP-ribose and GMP decreased APD50 at 26±7
%, 23±8 %, 26±8 % and 26±5 %; APD90 at 13±3 %, 10±3 %, 12±3 %, and 13±4 %, respectively. In
ground squirrel atrium adenosine and AMP decreased APD50 at 18±3 % and 10±3 % respectively;
APD90 at 11±2 % and 9±2 %, respectively. ADP-ribose and GMP did not reduce APD. Effects of
purine compounds in the rat heart were similar, except decrease of APD90 in ground squirrel PM
was up to 2–2.5 fold less then that in the rat PM. This peculiarity may contribute to insusceptibility
of ground squirrel myocardium to arrhythmias in presence of compounds that shorten APD (such as
adenosine).

P04
Chronic administration of serotonin transporter inhibitor (fluoxetine) decreases
monocrotaline-induced pulmonary hypertension in rats
KOZHEVNIKOVA VV, MEDVEDEVA NA
Biology Faculty of Moscow State University, Department of Human and Animal Physiology,
Leninsky Gory 1/12, Moscow 119992, Russia

Background: Pulmonary hypertension (PH) is characterized by high pulmonary blood pressure,
vascular remodeling, and right ventricular hypertrophy. Suggest that one of pathogenesis factor of
PH is the increase in plasma serotonin concentration. But the pathogenesis of PH is not fully
understood and the treatment of PH is still unresolved. Objective: The purpose of this study was to
investigate whether inhibition of 5- HT transporter by fluoxetine would prevent the development of
monocrotaline (MCT)-induced PH in rats. Methods: Daily supplementation with fluoxetine (10
mg/kg/day for 3 weeks) or vehicle was started 7 days after to a single-dose injection of MCT (60
mg/kg). On day 28, dose- dependent change of the reactivity of isolated pulmonary vessels, right
ventricular hypertrophy, and medial wall thickness were assessed. Results: In rats that received daily
supplementation of fluoxetine, vasoconstrictive response of pulmonary vessels with respect to

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                                             Abstracts

serotonin was significantly reduced (7.2±2.9 vs. 20.2±3.1). Medial wall thickness (24.03±1.35 vs.
36.08±1.52) of pulmonary vessels and right ventricular hypertrophy (33.0±1.7 vs. 36.2±1.2) of
fluoxetine-treated rats were less severe in rats with MCT-induced PH. Conclusions: We conclude
that daily supplementation of fluoxetine potently attenuates MCT-induced PH, right ventricular
hypertrophy, and vascular remodeling in rats. The results suggest that the chronic administration of
fluoxetine can restore the reactivity of pulmonary vessels and eliminate the symptoms of PH.

P05
Gonadectomy attenuates hypoxia-induced pulmonary hypertension in male rats
KUDRYAVTSEVA OS, MEDVEDEVA NA
Biology Faculty of Moscow State University, Department of Human and Animal Physiology,
Leninsky Gory 1/12, Moscow 119992, Russia

Objective: Gender-related differences in cardiovascular diseases, including pulmonary hypertension
(PH) may be mediated by androgen action on the vasculature. Design: twenty male Wistar rats were
gonadectomized at the age of 8 weeks and twenty rats were used as matched controls. 3 weeks after
operation was performed the rats were divided into 4 experimental groups: 1) normoxic rats with
intact testicles 2) hypoxic rats with intact testicles 3) normoxic gonadectomized rats 4) hypoxic
gonadectomized rats. Animals designated for exposure to CH were housed in a hypobaric chamber
at simulated altitude of 5 000 m, 10 h a day, 2 wk, 7 days/wk. Age-matched normoxic control
animals were housed at ambient air. After exposure to hypoxia, the right ventricular systolic pressure
(RVSP) and the right ventricular to total ventricular weight ratio (RV/T) and mean arterial pressure
(MAP) were measured as indices of PH. Results: No difference between measured variables was
detected in normoxic groups. Two weeks after hypoxic exposure only rats from 2 group developed
right ventricular hypertrophy (RV/T increased to 0.34±0.01 vs. 0.28±0.01 in 1 group) and systemic
hypertension (MAP increased to 119.8±2.1 mmHg vs. 95.7±3.7 in 1 group). Gonadectomized rats
showed no significant changes in RV/T and MAP compared to control. RVSP increased in both
groups, but it was significantly higher in intact hypoxic rats compared to gonadoectomized hypoxic
rats. Conclusion: gonadectomy leads to reduction of pathological changes associated with hypoxia-
induced PH both in pulmonary and systemic vasculature in male rats.

P06
Endothelin converting enzyme inhibitor decreases endothelin-1 plasma level and increases NO
production with urine in rats with hypoxia induced pulmonary arterial hypertension
SIMONOVA AI, POZDNEV VF, GOMAZKOV OA, MEDVEDEVA NA
Biology faculty of Moscow State University, Department of Human and Animal Physiology, Leninski
Gory 1/12, Moscow,119899 Russia

The significant role of the endothelium-derived relaxing factor nitric oxide (NO) and the
endothelium-derived vasoconstrictor peptide endothelin-1 (ET-1) in pathogenesis of pulmonary
arterial hypertension (PAH) is accepted. It is know that synthesis of endothelin is inhibited by NO,
and that NO plays a significant role in vascular tone regulation during hypoxia. Objective: The aim
of our study was to investigate the effects of endothelin converting enzyme inhibitor (ECEI) - PP36
(Pozdnev et al.1998) on NOx production with urine and endothelin-1 plasma levels in Wistar rats
with hypoxia induced PAH (hPAH). Methods: Animals were tested on tolerance to hypoxia and
were divided in 2 groups: low hypoxia tolerance (LHT) and high hypoxia tolerance (HHT). hPAH

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                                             Abstracts

was induced by the adaptation to hypobaric hypoxia (10% O2) during 2 weeks. PP36 (1.4 mg/kg)
was added to the drinking water during 2 weeks too. Urine NOx and ET-1 and angiotensin plasma
levels were measured. Results: hPAH increased production of NOx with urine in rats with HHT
(67.9±10.2 vs. 24.2±4.4) and decreased it in rats with LHT (16.3±0.4 vs. 24.2±4.4). ECEI increased
urine NOx in rats with hPAH and HHT (107.1±12.1 vs. 24.2±4.4) and showed no effect on rats with
LHT (15.9±0.5 vs. 24.2±4.4). Plasma levels of ET-1 in rats with hPAH and HHT was increased
(0.63±0.06 vs. 0.54±0.01).and 0.81±0.01 vs. 0.54±0.01 in rats with LHT. ECEI decreased ET-1
plasma level (0.42±0.02 vs. 0.52±0.01) in rats with hPAH both HHT and LHT. Conclusion: This
data shows that chronic treatment with ECE inhibitor significantly reduces endothelin-1 plasma level
in rats with hPAH. Changes in NOx production due to hPAH correlated with animals‟ tolerance to
hypoxia.

P07
Characterization of ecstasy (MDMA)-induced blood pressure, heart rate and respiratory rate
changes in anaesthetized female rats
SRISAWAT R, PUENGPAI S, NONTAMART N
School of Biology, Institute of Science, Suranaree University of Technology, 111 University Avenue,
Suranaree District, Amphur Muang, Nakhorn Ratchasima Province 30000, Thailand

In human, ingestion of ecstasy (MDMA) has been associated with acute adverse reactions such as
hyperthermia, hypertension and tachycardia. Little is known about what exactly MDMA does to
those physiological responses. We, therefore, investigated acute effects of MDMA on cardiovascular
and respiratory responses in female rats. Invasive arterial blood pressure, heart rate and respiratory
rate were recorded in pentobarbital-anaesthetized female rats for 2 hours after single i.v. injected
with vehicle (n=6), 5 mg/kg MDMA (n=7) and 10 mg/kg MDMA (n=8). Intragroup comparison
showed that MDMA markedly increased mean arterial blood pressure (MABP) and systolic blood
pressure immediately after MDMA administration (p<0.05, compared to baseline). In comparison
between groups, significant and non-dose-dependent increases in MABP, systolic blood pressure and
diastolic blood pressure can be observed immediately, 1 minute and 2 minutes after MDMA
(p<0.05, two way repeated measures ANOVA). However, there were no changes in those parameters
over the rest period of investigation. Heart rate was significantly increased 2, 3, 4 minutes after
MDMA (p<0.05, compared to control). Interestingly, significant increase in respiratory rate was
found in 10 mg/kg MDMA group at 2, 5, 10, 15 and 20 minutes after MDMA, whereas significant
decrease was observed in 5 mg/kg MDMA group at 70, 80, 105 and 120 minutes after MDMA
(p<0.05, compared to control). In conclusion, this study provided the first evidence of transient
cardiovascular response (increases in MABP, systolic blood pressure, diastolic blood pressure and
heart rate) and prolonged effects on respiratory response following acute MDMA in anesthetized
female rats.




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                                            Abstracts


            Poster session 02 – Cardiovascular physiology – peripheral mechanisms

P08
Vessel permeability of a macromolecular contrast agent in mouse muscle using Magnetic
Resonance Imaging (MRI)
RYGH CB, MOEN I, LØKKA G, TAXT T, SALVESEN G, REED RK, CURRY FR
Department of Biomedicine, University of Bergen, Jonas Lies vei 91, 5009 Bergen, Norway

Advances in MRI allow studies of physiological processes in time. Here we investigated vessel
permeability (Ps) in mouse muscle to a macromolecular tracer using MRI. Gadomer (kindly
provided by Bayer Schering Pharma, Germany), a gadolinium based contrast agent with an apparent
MW of 35 kDa, was injected into the tail vein of C57 black mice (25-30 g) that were anaesthetized
(Isoflurane, Isoba®vet, Schering-Plough, UK). The MR signal intensity (SI) was measured before,
during and after injection of the tracer using a T1-weighted dynamic sequence (TR/TE= 11.1/2.5 ms,
flip angle= 25°, time resolution 0.8 s, frames = 500). Regions of interests (ROIs) were placed over
the masseter muscle and skin to obtain dynamic information. The arterial input to muscle tissue was
estimated by placing small ROIs over supplying arteries to the tissue. The data analysis for
quantification of the microvessel permeability was based on the time-course of the tracer
concentration: Controlled bolus injection of 0.1 mmol Gd/kg BW caused an initial “step” increase in
tracer intensity as the vascular volume was filled. Subsequent blood to tissue tracer exchange
resulted in further (initially linear) increase (slope) in tracer intensity over periods of 100-300
seconds. Ps to Gadomer in tissue ROIs was calculated from the slope and step, assuming a
microvessel volume to surface ratio of 2.5 microns (Curry et al.1983). Average Ps values (× 10-7
cm/sec) were 2.9±1.0 (muscle) and 15±5 (skin). Corrections for the fall in plasma concentration
increased estimates by 10–20 %. The method enables non-invasive repeated measurements Ps in
organs of individual mouse of periods of days to weeks. References: Curry F.E., Huxley V.H. &
Adamson R.H. 1983. Am J Physiol, 245: 495-505.

P09
Role of cyclooxygenase-2 for lipopolysaccharide-mediated vasorelaxation in vitro and blood
pressure decrease in vivo
STAEHR M, HANSEN PB, MADSEN K, SKØTT O, JENSEN BL
Institute of Medical Biology, Physiologi and Pharmacology, J B Winsløws Vej 21, 5000 Odense,
Denmark

We hypothesized that lipopolysaccharide (LPS)-induced suppression of vasoreactivity is caused by
increased vascular COX-2 and downstream activation of eNOS by PGE 2 in the vascular wall. To
address this, chronic indwelling catheters were placed in the femoral artery and vein in mice for
blood pressure measurement and infusion, respectively. Five days after surgery, blood pressure and
heart rate were measured in conscious mice. The resting mean arterial pressure was 105.7±7.1
mmHg and heart rate 627.5±52.6 bpm. An LPS bolus (055:B5, 2 mg/kg, i.v.) reduced blood pressure
significantly after 4h (82.9±7.9 vs. control) and after 6h blood pressure was 79.3±3.6 mmHg. Heart
rate was significantly reduced 2 h after LPS (569.8±70.3) and after 6 h heart rate was 315.1±50.6
(n=11). Blood pressure and heart rate were not significantly different in COX-2 -/- mice (n=8)
compared to wild type (+/+) mice after LPS (n=4). Aorta-rings from COX-2 (+/+) and (-/-) mice
were incubated with LPS (055:B5, 50 g/mL, 18h, 37 ˚C) in cell culture medium and isometric


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                                             Abstracts

tension was recorded with a myograph (2 × 10 -7 M Phenylephrine (PE), 1h). Absence of functional
COX-2 in the vessels did not alter the effect of LPS on vasoreactivity in response to PE. We
conclude that COX-2 is not critical for acute blood pressure decrease or depressed vascular function
in murine models of LPS endotoxemia.

P10
Hypotensive effect of oxatriazolium-5-olate derivative in spontaneous hypertensive rats (SHR)
during chronic introduction
ARTEMIEVA MM, AKINFIEVA OV, POSTNIKOV AB, DALIGER IL, SHEVELEV SA,
MEDVEDEV OS, MEDVEDEVA NA
Moscow State University, Biology faculty, Department of human and animal physiology, Vorobievi
Gory, 1, Moscow, Russia

Soluble guanylate cyclase (sGC) is a crucial enzyme at NO/cGMP-mediated vasodilation. There are
NO-independent mechanisms of sGC activation besides wellknown enzyme activation by NO. Since
1966 oxatriazolium-5-olate derivatives are known as hypotensive agents at narcotized animals (Kier
LB et al., 1966). But the mechanism of their activity is not clarified. To examine hypotensive
activity of novel oxatriazolium-5- olate derivative AS-6 it was administrated per os during 3 weeks
in dose 5 mg/kg twice a day to male SHR rats (200-300 g weight). Control group was administrated
with appropriate water solutions. Systolic pressure was measured every 5 days by tail cuff. To
examine mechanism of hypotensive effect of AS-6 it was used preparation of isolated caudal artery.
It were perfused by AS-6 (6 × 0-5 М) and sodium nitroprusside solutions (1×10-6 М) in
phenylephrine (5×10-6 М) and phenylephrine (5×10- 6 М)+1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-
1- one (ODQ) (3×10-6 М) tonus. Oral administration of AS-6 in dose 5 mg/kg twice a day during 21
days leads to systolic MAP decrease in SHR (10.5±2.5 % in experiment, 4.1±1.9 % in control). Both
AS-6 and SNP produced decrease of perfusion pressure (p<0.05) in caudal artery of experimental
(63.2±4.0 % and 71.9±5.0 % respectively) and control (60.7±5.5 % and 68.8±6.4 %, respectively)
animals. ODQ significantly decreases vasodilatation caused by AS-6 and SNP both in experimental
(8.4±2.3 % and 15.2±2.8 %, respectively) and control (13.9±6.6 % and 30.5±8.6 %, respectively)
animals. So AS-6, as SNP, relaxes smooth muscle cells through activation of NO/sGC/cGMP-
dependent mechanism. In vivo experiments showed that AS-6 has prolonged (14 days) hypotensive
effect in SHR awake rats.

P11
The role of mitogen-activated protein kinase in the contraction of rat saphena artery
GAINULLINA DK, KALENCHUK VU, DIETTERLE VY, TARASOVA OS, VOROTNIKOV AV
M.V. Lomonosov Moscow State University, Leninskie Gory 1/12, 119991, Moscow, Russia

The increase in intracellular Ca2+ is a key stimulus for contraction of vascular smooth muscle
(VSM). Along with that, a number of intracellular signaling pathways sustain contraction without
additional rise of in Ca2+, thereby providing Ca2+-sensitisation of the contraction. Earlier our
colleagues demonstrated that Ca2+-sensitivity of VSM contraction in newborn rats is higher than in
adults, but the mechanism of such phenomenon has not been explored. Mitogen-activated protein
kinases (MAPK) are the molecules that take part in Ca2+- sensitisation of VSM contraction. We
investigated the role of two members of MAPK family (ERK1/2 and p38) in saphenous artery of 2-
week-old (2WK) and adult (AD) rats. For this purpose we studied the effects of ERK1/2 and p38

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                                             Abstracts

inhibition with U0126 and SB202190 respectively (10-5 M for both) on isometric contractile
response to methoxamine (selective α1-adrenoceptor agonist). The effect of SB202190 was greater
compared with U0126 and more prominent in 2WK than in AD. Together, the inhibitors decreased
maximal force in 2WK by about 70 % and in AD only by about 30 %. As the second step we studied
the degree of MAPK phosphorylation in relaxed state and after contraction with methoxamine (10-5
M). The segments were snap-frozen in liquid N2 and homogenized. Proteins separated with SDS-
PAGE were transferred to PVDF membranes. Membranes were incubated with anti-MAPK and anti-
phospho- MAPK antibodies and then visualized with enhanced chemiluminescence. The degree of
MAPK phosphorylation was nearly the same in 2WK and AD, but total MAPK content in 2WK was
greatly higher than in AD (3-fold for ERK1/2 and 5-fold for p38). We suggest that ERK1/2 and p38
pathways are more important for VSM contraction in young rats and may take part in Ca2+-
sensitisation.

P12
The role of Rho-kinase in adrenergic vasopressor response decreases during postnatal
development
TARASOVA NV, MOCHALOV SV, TARASOVA OS
Biological Faculty, M.V. Lomonosov Moscow State University, Leninskiye Gory, 1/12, Moscow,
119991, Russian Federation

Contraction of the vascular smooth muscle can be regulated by either Ca2+ signal or Ca2+-sensitivity
alteration. Rho- kinase (ROCK) plays an important role in the regulation of Ca2+-sensitivity. ROCK
inhibitors in vitro reduce vasoconstriction more considerably in newborn rats than in adults.
However this problem was not investigated in vivo. Present study was thus designed to compare the
effect of ROCK inhibitor fasudil (FAS) on the vasopressor response to alpha-1 adrenoceptors
agonist methoxamine (MX) in newborn and adult rats. Experiments were performed on 1-, 2- and 5-
6-week-old Wistar rats, anesthetized with urethane (1.2 g/kg). Catheters were implanted into the
right carotid artery and right jugular vein. Influence of FAS (3 mg/kg) on the response to MX (200
μg/kg bolus injection or 400 μg/kg infusion) was studied. Similar experiments were carried out after
autonomic blockade (chlorisondamine 2.5 mg/kg). FAS decreased baseline blood pressure (BP) by
10–15%, but did not change BP after autonomic blockade. FAS attenuated responses to MX in
newborn rats more considerably than in adults. FAS did not alter the maximum of MX response but
reduced its duration. Under autonomic blockade FAS reduced half-decay time 3-fold in adults and
almost 30-fold in newborn rats. During MX infusion integrated response for 6 min from the start of
infusion decreased by 65% in 1-week- old, by 27% in 2-week-old rats and by only 13% in adults.
FAS also markedly blocked vasoconstriction in newborns in the experiments on the isolated
saphenous artery. Thus we for the first time observed higher effect of ROCK inhibitor in vivo on the
pressor response to MX in rats during early postnatal development. This work was supported by
RFFI (grant 07-04-01527).




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                                             Abstracts


                    Poster session 03 – Cells, receptors, channels, membranes

P13
Biochemical characterization and comparison of N-terminal Gly16Arg variants of the 2-
adrenergic receptor
LILJA M, HAKALAHTI A, PETÄJÄ-REPO U
University of Oulu, Institute of Biomedicine, Department of Anatomy and Cell Biology, Oulu,
Finland

The 2-adrenergic receptor (2AR) carries two non- synonymous single nucleotide polymorphisms
in its extracellular domain that have been suggested to have an influence on the phenotype of
common disorders such as asthma. The 2ARs are widely studied members of the G protein-coupled
receptor family but little is known about their biosynthesis. In this study human 2AR Gly16Arg
variants were investigated in a heterologous expression system. Inducible HEK-293 cell lines stably
expressing the variants with an N-terminal Myc-tag and with or without a C-terminal Flag-tag were
used. Cell surface expression of the Arg16 variant was lower compared to the Gly16 variant when
measured by flow cytometry. By cell surface biotinylation and enzymatic deglycosylation assays,
followed by Western blot analysis, two receptor species were identified: a mature cell surface
receptor and an intracellular biosynthetic intermediate. Both variants were fairly efficient in
maturation as 70-80 % of the newly synthesized receptors were converted to mature receptors
studied by pulse-chase labeling. In addition, the two variants behaved similarly in terms of
maturation kinetics. However, after longer chase times the Arg16 variant was detected also in
smaller molecular weight forms that were absent for the Gly16 variant. They possibly represent
proteolytically cleaved fragments. Thus, a single amino acid change from a neutral glysine to a
charged arginine in the N-terminal region of the 2AR seems to decrease the cell surface expression
of the receptor. Since no significant differences in maturation were observed, these data indicate
faster turnover rate for the Arg16 variant at the cell surface.

P14
Regulation of cardiac activity by Kir2.x inward rectifier potassium channels in fish
PAAJANEN V, HASSINEN M, VORNANEN M
Faculty of Biosciences, University of Joensuu, Joensuu, Finland

Inward rectifier potassium channels (Kir2.x) in cardiac myocytes have an essential role in thermal
plasticity of ectothermic animals as regulators of resting membrane potential and modulators of
action potential duration. They form functional channels as homo- and heterotetrameres allowing
potassium current (IK1) with channel type and intracellular rectification factors (IRFs) dependent
rectification properties. We have tested the role of Kir2.x channels by using electrophysiological and
molecular biology techniques and by comparing functions of endogenous ion channels of fish
(crucian carp, Carassius carassius, and rainbow trout, Oncorhynhus mykiss) and recombinant fish
channel proteins expressed in mammalian cell lines. We have also used electrophysiological and
molecular modeling to characterize the physiological importance and structure related functionality,
respective. We have cloned 7 members of Kir2 family from fish including a novel Kir2 protein:
ccKir2.5. The novel channel is the predominant IK1 forming channel in cold acclimated crucian carp
and has the steepest inward rectification among Kir2 family. Furthermore, we have showed with
quantitative RT-PCR and single channel patch-clamp methods the acclimation dependency of

                                                 103
                                             Abstracts

expression of Kir2.x coding genes and functional proteins in both fish species. Our results suggest
that composition of IK1 mediating Kir2.x channels can be important for thermal plasticity of
animals. However, the mechanical explanation of seasonal switch between different Kir2.x channels
in cardiac myocytes remains to be elucidated.

P15
Electrophysiological properties fish Kir2.x channels expressed in mammalian cell lines
PEKURINEN T, PAAJANEN V
Faculty of Biosciences, University of Joensuu, Joensuu, Finland

Thermal acclimation modulates membrane proteins as well as their environment, membrane lipids, a
known modulator of several membrane proteins. Therefore, the structure-related functional
differences in ion channels of thermally acclimated animals can not be tested explicitly without the
use of recombinant ion channels expressed in controllable lipid environment. We have compared the
functional properties of recombinant and endogenous fish ion channels to separate the regulatory
properties mediated via the environment from the protein-structure based differences. We have
expressed members of Kir2 family in mammalian cell lines (COS-1 and HEK-293e) and compared
their electrophysiological properties and regulation with patch-clamp methods. We have tested
channel sensitivity to two conventional blockers, Ba2+ and TEA, both of which are thought to
interact with only with one – few residues in external part of the ion channel. Surprisingly, both
blockers inhibit the potassium current in a cell type dependent manner. For example, trout Kir2
proteins (omKir2.1 & omKir2.2) expressed in COS-1 cells can be blocked with micromolar
concentrations of external barium whereas much smaller concentrations are needed to block the
channels in fish cardiac myocyte (IC50 from 0.2 to 25 µM). Similar differences exist also in TEA-
sensitivity (IC50 from 3 to over 100 mM). Our results demonstrate that a more suitable expression
system is needed for the future studies of fish membrane proteins. Beside giving physiologically
meaningful results from the isolated teleost membrane proteins the development of an endogenous-
like expression system can have interest in membrane – ion channel interactions in general.

P16
The effect of osmotic shock on the early mouse embryo volume
POGORELOVA MA, GOLICHENKOV VA, POGORELOVA VN
Biological Faculty, Moscow State University, Leninskie gory, Moscow, 117234, Russia, ITEB, RAS,
Pushchino, Moscow province, 142290, Russia

The early embryo membrane is highly permeable for water, that makes the cell very sensitive to
osmotic shock. The cell volume alteration plays an important role in regulation of key cellular
functions, including metabolism, protein synthesis and cell death. In the given research, the volume
changing of embryonic compartments were determined for isotonic and anisotonic conditions using
three- dimensional reconstruction (3-DR). The keeping of the intact volume of the 2-cell mouse
embryo was based on freeze-drying technique (Pogorelov & Pogorelova 2008). A Z-stack of optical
slices was obtained with laser scanning microscope (LSM) (Zeiss 510, Germany). Osmotic shock
was modeled by changes of NaCl contents in Dulbecco‟s medium. As control we considered the
embryo cryofixed immediately after flushing from oviduct. Its volume was calculated as (56±5) ×
103 µm3 (mean ± the standard deviation). Our data indicates that hypo-osmotic (70 mM NaCl)
incubation for 10 minutes resulted in the swelling peak of blastomere volume to (86±11) × 103 µm3.

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                                              Abstracts

After 60 minute incubation the cell volume was recovered to control level. A hyperosmotic (280 mM
NaCl) shock induced quick shrinkage of embryonic cells to (31±2) × 103 µm3 and this effect was
irreversible. Note, that incubation in conventional Dulbecco‟s medium caused a gradual decrease of
the blastomere volume to (42±5) × 103 µm3. The developed technology of LSM tomography of
mouse early embryo allows us to measure the cell volume corresponding to the life-like state. Our
quantitative data correspond to the qualitative effects observed in vitro. References: Pogorelov, A. &
Pogorelova, V. 2008. J.Microsc. 232.

P17
The antimicrobial peptide plantaricin A permeabilizes liver and kidney cells
ANDERSLAND K, JOHANSEN GF, HAUG TM, SAND O
Department of Molecular Biosciences, University of Oslo, PB 1041 Blindern, NO-0316 Oslo,
Norway

Antimicrobial peptides are produced by nearly all organisms: bacteria, plants, invertebrates, and
vertebrates. Certain antimicrobial peptides from multicellular animals also kill a variety of tumour
cells at concentrations not affecting normal eukaryotic cells. Recently, it was reported that also
Plantaricin A (PlnA), which is a peptide with strain-specific antibacterial activity produced by
Lactobacillus plantarum, may kill eukaryotic cells (Sand et al. 2007). It was shown that PlnA
permebilizes cancerous rat pituitary cells (GH4 cells), whereas normal rat anterior pituitary cells are
resistant to the peptide. In order to examine if the preferential permeabilization of cancerous cells is
a general feature of PlnA, we have studied its effect on primary cultures of rat liver cells
(hepatocytes and endothelial cells) and two epithelial cell lines of primate kidney origin (Vero cells
from green monkey and human Caki-2 cells). The Vero cell line is derived from normal cells,
whereas the Caki-2 cell line is derived from a cancerous tumour. The membrane effects were studied
by means of patch clamp recordings and microfluorometric (fura-2) monitoring of the cytosolic
concentrations of Ca2+ ([Ca2+]i ) and fluorochrome. In all the tested cell types, exposure to 10-100
µM PlnA induced a nearly instant permeabilization of the membrane, indicated by the following
criteria: increased membrane conductance, membrane depolarization, increased [Ca2+]i, and
diffusional loss of fluorochrome from the cytosol. At a concentration of 5 µM, PlnA had no effect on
any of the cell types. We conclude that the permeabilizing effect of PlnA is not restricted to
cancerous cells. Reference: Sand, S.L., Haug, T.M, Nissen-Meyer, J. & Sand, O. 2007. J Membrane
Biol 216, 61–71.

P18
Both normal and cancerous lymphocytes and neurons are permeabilized by plantaricin A, a
peptide produced by Lactobacillus plantarum
SAND SL, OHARA S*, OPPEGÅRD C, IIJIMA T*, BLOMHOFF HK**, NISSEN-MEYER J,
SAND O
Department of Molecular Biosciences, University of Oslo, PB 1041 Blindern, NO-0316 Oslo,
Norway

Antimicrobial peptides produced by multi-cellular organisms protect against pathogenic
microorganisms, whereas such peptides produced by bacteria provide an ecological advantage over
competitors. Certain antimicrobial peptides of metazoan origin also kill a variety of tumour cells.
Plantaricin A (PlnA) is a peptide with membrane-permeabilizing strain-specific antimicrobial

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                                              Abstracts

activity produced by Lactobacillus plantarum C11. Recently, we have reported that PlnA also
permeabilizes cancerous rat pituitary cells (GH4 cells), whereas normal rat anterior pituitary cells are
resistant (Sand et al. 2007). To investigate if the preferential effect on cancerous cells is a general
feature of PlnA, we have studied effects of the peptide on normal and cancerous lymphocytes and
neurons. The sensitivity to PlnA of normal human T- and B- cells, Jurkat cells (from human T-cell
leukaemia), and Reh cells (from human B-cell leukaemia) was studied by using flow cytometric
techniques to detect morphological changes. The membrane-permeabilizing effect of PlnA on
normal cortical neurons in primary cultures from embryonic rats, PC12 cells (postganglionic,
sympathetic neuron-related cells derived from a rat adrenal chromaffin tumour), and murine Neuro-
2A cells (derived from the C1300 spinal cord tumor) were studied by Ca2+- imaging using a
combination of fluo-4 and fura-red as fluorochromes. All the tested cell types were affected by 10-
100 µM PlnA, whereas concentrations below 10 µM had no detectable effect. We conclude that
PlnA permeabilizes normal neurons and lymphocytes, and various cancerous counterparts, at about
the same concentration. Reference: Sand, S.L., Haug, T.M, Nissen-Meyer, J. & Sand, O. 2007. J
Membrane Biol 216, 61–71.

P19
Determination of cell viability of mesenchymal stem cells by analyzing mitochondrial function
PIETILÄ M, LEHTONEN S, NORDSTRÖM K, LEHENKARI P
Institute of Biomedicine/Anatomy, University of Oulu, Oulu, Finland

Background. Cell based medicinal products involve risks, therefore, it is extremely important to
prove the safety of these innovative treatments. We have developed procedure to determine the
viability of human mesenchymal stem cells (hMSCs) by analyzing mitochondrial function thus
ensure the safety of hMSCs in future cell therapy treatments. Methods and results. FACS-based
method to monitor the inner membrane potential of mitochondria was established. The fluorescent
membrane-permeant cation, rhodamine123, was used in quench mode. Mesenchymal stem cells
were exposed to 2.6 M Rhodamine123 and 7AAD was used to detect dead cells. Mitochondrial
DNA (mtDNA) from hMSCs of different aged patients was analyzed by expand PCR to determine
the amount of deletions. Results have shown that there are variations in viability between hMSC
lines. The proliferation capacity of analyzed cells has correlated to the rhodamine123 intensity.
Analyzed cells can be categorized into three groups based on the proliferation capacity and
rhodamine123 intensity. Cells that proliferate rapidly have rhodamine123 intensity about 2-3 times
lower than cells that do not proliferate so actively. mtDNA deletions have been found from three
different patients, some were heteroplasmic small deletions and others homoplasmic larger deletions.
Conclusions. It is possible to distinguish cell populations with different proliferation capacities by
this FACS-based method. In future, the idea is to improve the sensitivity of this test. mtDNA
analysis have revealed interesting results (3 deletions from 17 patients) and needs more
investigations in future.




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                                              Abstracts


                        Poster session 04 – Endocrinology and metabolism

P20
Role of ACBP in food intake and hypothalamic gene expression in transgenic rats
OIKARI S, HUOTARI A, MAURIALA T, AURIOLA S, HEINONEN M, KIEHNE K, FÖLSCH
UR, KLAUSZ K, JÄNNE J, ALHONEN L, HERZIG K-H
Institute of Biomedicine/Physiology, University of Oulu, Finland, AIV-Institute, University of
Kuopio, PL 1627, 70211 Kuopio, Finland

Recent studies have indicated that long chain fatty acids, or their metabolic intermediates acyl-CoAs,
act as signals of body‟s energy state and influence the hypothalamic regulation of food intake. In
cytosol Acyl-CoAs are bound by proteins such as acyl-CoA binding protein (ACBP). This is
influencing the metabolic and regulatory functions of acyl-CoAs. We utilized transgenic (tg) rats
overexpressing mouse ACBP gene to study the role of ACBP on the hypothalamic regulation of food
intake. Overexpression of ACBP had no influence on the food intake or the body weight of rats.
Furthermore, ACBP overexpression did not affect brain acyl-CoA pool. In contrast, tg rats had
nutritional state specific changes in the gene expression profile. In fed tg rats FAS, SIRT-1 and
PPAR mRNA levels were reduced 28%, 26 % and 43 %, respectively, while the mRNA level of
CPT Ic was increased by 22 %. mRNA levels of SREBP-1 and protein levels of AMPK were equal
between fed tg and syngenic animals. Our results demonstrate that preventive measures abolish the
effects of the constant overexpression of ACBP. These include the observed changes in the gene
expression that prevent the increase of acyl-CoA pool size.

P21
Evaluation of association between leptin levels and leptin gene polymorphism in obese females
OKUDAN N, GÖKBEL H, ACAR H, UZUNOĞLU S
Department of Physiology, Meram Faculty of Medicine, Selcuk University, Konya, Turkey

Objective: The aims of the study were to determine serum leptin levels in obese and healthy females,
and to study whether there was a polymorphism in 25th codon of leptin gene. Methods: Eighty-
seven obese (BMI 40.1±3.9) and 75 healthy women (BMI 22.1 ±1.6) were constituted obese and
control groups. Body fat percent, fat mass, lean body mass and body water percent were determined
by bioimpedance meter. Serum leptin level was measured by ELISA method. Genomic DNA was
isolated and the presence of 25th codon polymorphism in the leptin gene was determined by PCR
technique. Results: Mean leptin level of the obese group (147.9 ±44.8 ng/ml) was significantly
different from those of the control group (38.5±22.0 ng/ml). The correlation with serum leptin level
to body fat percent and fat mass was significant in the control group, but not significant in the obese
group. No polymorphism in 25th codon of leptin gene in both groups was detected. CAA/CAA
pattern was observed in all samples. Conclusion: The difference of leptin levels between control and
obese groups can not be associated with the 25th codon polymorphism of the leptin gene. This data
implies that difference of leptin levels between control and obese groups are more likely to be
associated with the polymorphism in the leptin receptor gene.




                                                 107
                                             Abstracts

P22
Increased heat production and upregulation of PPARs and UCP2 mRNA In WAT in mice
overexpressing human prepro-orexin
MÄKELÄ KA, AHTIALANSAARI T, VÕIKAR V, SAKURAI T, ALHONEN L, HERZIG K-H
Institute of Biomedicine/Physiology, University of Oulu, Finland

Orexins are hypothalamic neuropeptides involved in feeding, energy homeostasis and vigilance.
They have also been shown to play important role in the periphery. Recently, orexin-A has been
reported to affect thermogenesis. Since the peripheral actions of orexins are still unclear, we
developed a transgenic (tg) mouse line overexpressing the human prepro-orexin (hPPO) gene under
the control of endogenous promoter. Transgenic mice were characterized using PCR, Southern and
Western blotting. Metabolic performance, home cage activity as well as drinking and feeding
behavior of 6 tg and 6 syngenic (sg) mice were measured using an automated monitoring system.
Total RNA was extracted from white and brown adipose tissue (WAT/BAT) and skeletal muscle
from fed mice and reverse transcribed to cDNA. Expression of uncoupling proteins (UCP) 1-3 and
peroxisome proliferator-activated receptors (PPARs delta and gamma) were analyzed by quantitative
real- time PCR. Our tg animals showed significantly increased expression of hPPO/orexin-A.
Interestingly, orexin-A was expressed also in WAT indicating a role of orexins as an adipokine.
Preliminary evaluation documented a similar food/drink intake and locomotor activity between our
animals. However, we found a significant effect on heat production in tg animals. Expression levels
of PPARs and UCP2 in WAT were significantly increased in tg mice compared to sg littermates. In
conclusions, orexins can regulate heat production and mRNA expression of PPARs as well as UCP2
in WAT thus providing new insights for regulation of body homeostasis.

P23
Regulation of visfatin expression by feeding status in mice
HUOTARI A, KYRYLENKO O, STÜTZER I, PURHONEN AK, WALKOWIAK J, HERZIG K-H
Institute of Biomedicine/Physiology, University of Oulu, Finland

Visfatin is a novel member of the adipokine family and is identical to nicotinamide
phosphoribosyltransferase (Nampt), an essential rate-limiting enzyme in biosynthesis of
nicotinamide adenine dinucleotide (NAD+). Regulation of visfatin remains largely unknown. To
study the regulation of visfatin in response to nutritional state, we examined expression levels of
visfatin mRNA in fasted and fed mice. 3-month-old male C57BL/6J mice were fed ad libitum, or
fasted 18 hrs and sacrificed. The mRNA expression was analyzed by real-time PCR and protein
levels were assessed by Western blotting. Visfatin mRNA in brown adipose tissue (BAT) was 10-
fold higher compared to liver (p≤ 0.001) and 2,5-fold higher than in visceral adipose tissue (vWAT).
Visfatin mRNA was 2-fold higher in vWAT than in subcutaneous (scWAT). The expression of
visfatin in BAT was confirmed on protein level (4,8-fold higher compared to liver, p≤0.01 ). Visfatin
mRNA in BAT and liver was significantly increased by 2-fold and 1.6 fold (p≤0.01 in both),
respectively, after 18 hrs fasting, while mRNA in vWAT or scWAT did not change compared to fed
mice. Visfatin is highly expressed in brown adipose tissue in mRNA and protein levels and visfatin
mRNA is upregulated in liver and BAT by fasting. As visfatin is a regulator of NAD+ synthesis, the
expression of visfatin in BAT might suggest a novel role of visfatin in the regulation of energy
metabolism and thermogenesis.



                                                108
                                             Abstracts

P24
Effects of reported previous physical activity on body composition and its changes during
military service
MÄKINEN TM, MIKKOLA I, JOKELAINEN J, TIMONEN M, HÄRKÖNEN P,
SAASTAMOINEN E, LAAKSO M, PEITSO A, JUUTI A-K, KEINÄNEN-KIUKAANNIEMI S
Institute of Health Sciences, University of Oulu, P.O. Box 5000, FIN-90014 University of Oulu,
Oulu, Finland

Objectives: The objective of the study was to examine how reported previous physical activity
affects body composition of Finnish young men during military service, which is associated with
marked changes in diet and physical activity. Methods: Altogether 1022 men (19 yrs) were followed
throughout their military service (6-12 months). Height, weight, waist circumference and waist-to-
hip (WHR) ratio were recorded. Body composition was measured by bioelectrical impedance
assessments (BIA) in the beginning and end of the service. The measured parameters were fat mass
(FM), fat percent, fat free mass (FFM) visceral fat area (VFA), lean body mass and muscle mass.
Previous physical activity was assessed in the beginning of the service by a questionnaire. Results:
On average military training decreased weight 0.6%, reduced WHR 1.4%, FM 10.0%, fat percent 6.0
%, and VFA 43.3%. FFM increased by 1.4%, lean body mass 1.3% and muscle mass by 1.6%. The
group of under- and normal weight men gained weight, FM and FFM, whereas overweight and
obese men lost weight, FM and gained FFM. Fat tissue was reduced most in the groups of
overweight (20%) and obese (25%) men. The amount of visceral fat was reduced in all BMI groups
(38-44%). Low physical activity before military service was reported most often in obese subjects
(32%), and high physical activity most frequently in normal weight men (61%). Among overweight
men who reported being inactive previous to the military service weight, WHR, BMI, FM, fat% and
VFA was reduced more compared with those who reported more physical activity. Conclusions: The
lifestyle changes associated with military service markedly reduce fat tissue and increase the amount
of lean tissue. These beneficial changes are especially prominent among previously inactive young
men with high BMI.

P25
Effects of coenzyme Q10 supplementation on plasma adiponectin, interleukin-6 and tumor
necrosis factor-alpha levels in males
GÖKBEL H, GERGERLİOĞLU HS, OKUDAN N, GÜL İ, BÜYÜKBAŞ S, BELVİRANLI M
Department of Physiology, Meram Faculty of Medicine, Selçuk University, Konya, Turkey

Objective: The aim of the study was to determine the effects of coenzyme Q10 supplementation on
plasma adiponectin, interleukin-6 and tumor necrosis factor-alpha levels in sedentary males.
Methods: Fourteen healthy, non-smoker and sedentary males participated in the study. The protocol
was approved by the Ethical Committee of the Faculty. This study was a randomized, double blind,
crossover trial. From all participants blood samples were collected before coenzyme Q10 or placebo
supplementation. The participants were randomly allocated to two groups. Seven participants
received oral coenzyme Q10 supplementation, seven participants received placebo (glucose) for 8
weeks. At the end of the 8-week, second blood sampling was performed. After a 4-week washout
period, placebo was given to the participants who used coenzyme Q10 first time and vice versa and
blood sampling was repeated. Plasma samples were stored at -80°C until the time of analysis for
adiponectin, interleukin-6 and tumor necrosis factor-alpha. Results: Neither coenzyme Q10 nor

                                                109
                                              Abstracts

placebo supplementation did affect plasma adiponectin and tumor necrosis factor-alpha levels.
Interleukin-6 level increased with coenzyme Q10 supplementation but this increase did not different
from that seen with placebo supplementation. Conclusion: Coenzyme Q10 supplementation did not
affect plasma adiponectin, interleukin-6 and tumor necrosis factor-alpha levels in sedentary males.

P26
Identification of possible ovarian cancer specific biomarkers in the tumour interstitial fluid
HOX H, TENSTAD O, KOLMANNSKOG O, WOIE K, SALVESEN H, WIIG H
Department of Biomedicine, University of Bergen, N-5009 Bergen, Norway

Ovarian cancer is the most lethal gynaecological malignancy, with the majority of cases diagnosed
with metastatic disease. If there are tumour specific biomarkers present in plasma at an early stage of
the disease, these might contribute to an earlier detection and therapy and potentially a more positive
outcome. Proteins specific for the tumour tissue can be expected to be found in high concentrations
in the interstitial fluid compared to plasma, suggesting that the tumour interstitial fluid may be a
valuable source for identification of tumour specific proteins. Thus, we have isolated the tumour
interstitial fluid from human ovarian cancer tissue by centrifugation, i.e. by exposing tumour
specimens to 38 g as described by Wiig et al. 2003. To allow a better separation and identification of
low abundant proteins the fluid was further fractionated by immunodepletion and reversed phase
chromatography. This fluid was compared to plasma from the patients and healthy women with
respect to protein composition in one-dimensional liquid chromatography coupled with tandem mass
spectrometry. Eighty-seven and 91 proteins were identified by at least two peptides in control
plasma and patient plasma respectively, and 232 proteins were identified by at least two peptides in
tumour interstitial fluid. Our experiments show that tumour interstitial fluid is a rich source for
tumour specific proteins, and some of these may be candidates for the use in ovarian cancer
diagnostics. Reference: Wiig, H., Aukland, K. & Tenstad, O. 2003. Am J Physiol Heart Circ Physiol
284, H416-H424.

P27
The estimation of milk intake and energy expenditure of reindeer calves by the doubly-labelled
water method
SOPPELA P, TIMLIN S, VISSER H, NIEMINEN M
Arctic Centre, University of Lapland, PO Box 122, FIN-96101 Rovaniemi, Finland

Milk intake and energy expenditure was measured by doubly-labelled water (DLW, 2H218O) in
reindeer calves during their first weeks of life. Two successive experiments (each 7 days) were
conducted in four calves at the ages of 1-2 and 3- 4 weeks. The calves were freely suckling their
mothers which were fed high-protein concentrates. The milk intake of the calves was on average
1.28 kg/day (1.04-1.50 kg/day) during the first experiment and 1.47 kg/day (1.23-1.88 kg/day)
during the second experiment. The energy expenditure of the calves was on average 5.87 MJ/day at
the age of 1-2 weeks and 7.42 MJ/day at the age of 3-4 weeks. The milk output of the mothers,
measured by a milking machine, was on average 1.37 kg/day (0.88-1.80 kg/day) at the end of the
first experiment and 1.12 kg/day (0.52- 1.74 kg/day) at the end of the second experiment. Milk
contained on average 10.8 % fat, 8.2 % protein, 4.5 % lactose, 23.7 % dry matter and 6.9 kJ/g gross
energy. The body weight gain of the calves correlated with the milk intake and energy expenditure.
The milk intake correlated also with the body weight and feed intake of the mothers. The results

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                                              Abstracts

show that DLW method suits for the measurement of milk intake of reindeer calves during the peak
suckling period when their major water source is milk. The benefit of the method is that the lactation
remains undisturbed and the energy expenditure can be measured simultaneously. The disadvantage
is that the method is technically demanding and expensive.




                                                 111
                                              Abstracts


                       Poster session 05 – Muscle and exercise physiology I

P28
The use of a cooling vest during exercise in warm condition delays the appearance of fatigue
and improves cycling performance
DUGUE B, LUOMALA M, HOLMER I, SMOLANDER J, OKSA J
Faculty of Sport Sciences, 4 allée Jean Monnet, Poitiers, France

Whether the use of a cooling vest during exercise in warm and humid environments promotes
endurance is not known. Here we address this question for heavy cycling. Seven trained males were
evaluated 3 times: first to determine their maximal capacities and twice during heavy cycling with or
without using a light cooling vest (Flexi cold vest, Sweden, ice packs at -20 °C). After a standardised
warm-up, the subjects followed a cycling protocol at 30°C and 40% relative humidity. The subjects
cycled at 60% VO2 max until exhaustion except for periods of 1 min starting after 9 min and every
10 min thereafter when cycling was at 80% VO2 max. The cooling vest was put on after 30 min. The
time to exhaustion and rectal temperature after 50 min of cycling and at exhaustion and EMG-
signals from 4 muscles of the right leg were measured. We found that the time to exhaustion was
significantly improved when the vest was used compared with the control session (72±17 min vs. 58
±7 min, respectively). After 50 min of cycling the rectal temperature did not differ between the two
sessions whereas at exhaustion it was significantly higher in the session where the vest was used
compared with the control session (39.1 vs. 38.8°C, respectively). Also, the frequency and the power
analysis of the EMG signals indicated lower neuromuscular fatigue when the vest was used. In
conclusion, wearing an ice-vest while exercising in warm conditions improves endurance
performance and delays the appearance of fatigue.

P29
Effects of menstrual cycle, oral contraception, and training on exercise-induced changes in
circulating DHEA-sulphate and testosterone in young women
ENEA C, BOISSEAU N, OTTAVY M, MULLIEZ J, MILLET C, DIAZ V, DUGUE B
Faculté des Sciences du Sport, EA 3815, Université de Poitiers, France

Aim: We investigated the effects of menstrual cycle, oral contraception, and training status on the
exhaustive exercise-induced changes in circulating DHEA-sulphate and testosterone in young
women. Methods: Twenty-eight healthy women were allocated to an untrained group (n=16) and a
trained group (n=12) depending on their training background. The untrained group was composed of
9 oral contraceptive users (OC+) and 7 eumenorrheic women (OC-). The trained group was
composed of OC+ subjects only. All the OC+ subjects were taking the same low-dose oral
contraception. Three laboratory sessions were organised in a randomised order: a prolonged exercise
test until exhaustion, a short-term exhaustive exercise test, and a control session. Blood specimens
were collected before, during and after the exercise tests and at the same time of the day during the
control session. Results: Basal circulating testosterone was significantly lower in trained than in
untrained subjects. In untrained OC- and trained OC+ subjects, the prolonged exhaustive exercise
induced a significant increase in circulating DHEA-s and testosterone whereas the short-term
exercise induced a significant increase in circulating DHEA-s, only. However, the magnitude of the
changes tended to be smaller in trained OC+. No change was observed in untrained OC+ after both
types of exercise. Menstrual phases in OC- did not influence the responses. Conclusion: Exhaustive


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                                              Abstracts

physical exercise is able to induce an increase in circulating DHEA- sulphate and testosterone in
young women. Oral contraception may limit such exercise-induced changes. Acknowledgements:
The authors thank the World Anti- Doping Agency (WADA) for financial support.

P30
Soldiers' performance during a two-week field exercise in winter
RISSANEN S, OKSA J, MÄKINEN T, RINTAMÄKI H, PEITSO A
Finnish Institute of Occupational Health Oulu, Physical Work Capacity team, Aapistie 1, 90220
Oulu, Finland

The physical fitness of the young Finnish men called up to the military service has gradually
declined during the last three decades. During the 6-month military service conscripts undergo basic
military and physical exercise training which should improve their fitness. The objective of this
study was to investigate cardiorespiratory and muscular performance of the conscripts during their
last and longest (12 days) field exercise of the military service in the winter conditions. Hypothesis
is that if excessive fatigue exists during the prolonged training, it should be seen as reduced
performance capacity. Subjects were 21 male jaegers. Each subject participated three times in
maximal muscle performance and cardiorespiratory tests: three days before (T1), at the 5th day (T2)
and at the 12th day (T3) of the field training. Maximal isometric leg extension (LE), static jump (SJ),
counter movement jump (CMJ) and wrist rotation (WR) were tested. SJ, CMJ and WR were also
measured every day in the field. Mean maximal VO2 was maintained but maximal heart rate
significantly decreased in T3 compared to T1. No significant changes occurred in SJ, CMJ, LE and
WR in T2 or T3. Daily measurements after strenuous physical activity showed significant but
temporary decrement in muscle performance. Minor changes in cardiorespiratory and neuromuscular
performance suggest that the partly motorized infantry troops were capable to perform their field
training without cumulative fatigue. The results reflect that the physical training during the military
service is adequate to elevate the cardiorespiratory and muscular performance to the level which is
sufficient in order to endure prolonged field exercise in infantry troops.

P31
The swimming performance of brown trout and whitefish: the effects of exercise on Ca2+
handling and oxidative capacity of swimming muscles
ANTTILA K, MÄNTTÄRI S
Department of Biology, University of Oulu, Oulu, Finland

The swimming performance of two fish species having initially different swimming strategies,
brown trout and whitefish, was measured after training in order to relate the effects of exercise on
calcium handling and muscle performance of fish. The time to 50% fatigue was measured during the
training period, and compared with the density of dihydropyridine (DHP) and ryanodine (Ry)
receptors determined by histochemical analysis from swimming muscles. Overall, both trained
brown trout and whitefish had superior swimming performance as compared to control ones.
Interestingly, brown trout achieved the highest swimming performance swimming against the water
flow velocity of 2 BL/s while among whitefish the best efficiency was seen after training with lower
swimming velocities. Training also induced a significant increase in DHP and Ry receptor density in
both species. Generally, in brown trout the most notable increase was observed in red muscle
sections from the fish swimming for six weeks against water currents of 1 BL/s (DHPR 176.5±7.7%

                                                 113
                                             Abstracts

and RyR 231.4±11.8%) and white muscle sections against 2 BL/s (DHPR 129.6±12.4% and RyR
161.9±15.5%). In whitefish the most prominent alterations were noted in samples from both muscle
types after six weeks of training against water current of 1.5 BL/s (DHPR 167.1±16.9% and RyR
190.4±19.4%). To conclude, our findings demonstrate an improved swimming performance and
enhanced calcium regulation after training. Moreover, there seems to be a connection between the
swimming performance and receptor levels especially in white swimming muscles of different fish
species, regardless of their initially deviant swimming behaviours. However, depending on the
training regimen the divergent swimming behaviours do cause a different response.

P32
-Lipoic acid attenuates exercise-induced oxidative stress without impairing exercise capacity
KINNUNEN S, OKSALA N, HYYPPÄ S, JAKUS J, ATALAY M
Institute of Biomedicine/Physiology, University of Kuopio, Finland

While antioxidant supplementation apparently decreases exercise-induced oxidative stress, there is
also a risk of attenuating the physiological response of tissues to exercise and blunting the
adaptations. In this study we examined the protective role of natural thiol antioxidant, -lipoic acid
(LA) supplementation at rest and during recovery after an acute exercise in plasma and in skeletal
muscle of horse. Six standardbred trotters were examined on the treadmill and exercised 75 min at
individually defined aerobic level. They were supplemented orally with LA (25 mg kg-1 day-1) for
five weeks without any additional vitamins. Using electro paramagnetic resonance assay, we showed
that strenuous aerobic exercise increases significantly free radical formation in gluteus medius
muscle during control period but not after LA-supplementation. LA-supplementation decreased the
amount of post-exercise lipid hydroperoxides in plasma and exercise-induced increase in
malondialdehyde in plasma and muscle. There was no difference in muscle protein carbonyl levels; a
marker of protein oxidation. Furthermore, LA increased the oxidative capacity of the muscle
measured by citrate synthase activity and lowered the blood lactate concentration during exercise.
Our results suggest that LA-supplementation appears to decrease exercise-induced oxidative stress at
lipid phase without impairing the athletic capacity of the muscle in horse.




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                                              Abstracts


                      Poster session 06 – Muscle and exercise physiology II

P33
Endurance training exerts differential effects on diaphragm and gastrocnemius muscle feed
arteries in the rat
BORZYKH AA, KALENCHUK VU, ANDREEV-ANDRIEVSKII AA, BURAVKOV SV,
VINOGRADOVA OL
SRC RF - Institute for Biomedical Problems RAS (Khoroshevskoe shosse 76A, 123007, Moscow,
Russia), M.V. Lomonosov Moscow State University, Leninskie Gory 1/12, Moscow, 119991, Russia

During exercise muscle blood flow increases in proportion to the intensity of activity. In addition to
locomotory muscles, aerobic exercise inevitably results in high load on respiratory muscles. This
work aimed at testing the hypothesis that the effect of training on muscle blood vessels is also
dependent on the functional load on the muscle. Male Wistar rats were treadmill trained 6 days/wk
for 8 wk. Efficacy of training was confirmed by 15% increase of max O2 consumption compared to
sedentary control. Isometric contractile and relaxation properties of rings cut from diaphragm (DA)
and medial gastrocnemius muscle (GA) feed arteries were studied in vitro. Both arteries had a
diameter of 200-250 micron. In controls DA as compared to GA had greater density of sympathetic
innervation and higher sensitivity to noradrenaline (NA), but not to serotonin. After training
sensitivity to NA did not change in GA, but was reduced in DA; decrease of postsynaptic sensitivity
must weaken sympathetic constriction of DA during exercise. In controls dilator responses to
acetylcholine (Ach) and sodium nitroprusside (SNP) in DA were also greater than in GA. After
training reactivity to Ach was prominently increased only in GA, the effect was abolished after
blockade of NO-synthesis (L- NAME, 10-4M). Interestingly, reactivity of both arteries to SNP was
diminished by training indicating smaller sensitivity of smooth muscle to NO. Our results permit the
conclusion that differential effects of exercise training on DA and GA are related to basic differences
in vasomotor regulation of these arteries. The work was supported by the RFFI (grant 06-04-49699-
a).

P34
The reproducibility of knee cartilage magnetic resonance imaging with dGEMRIC method
ISSAKAINEN J, MULTANEN J, RAUVALA E, LAMMENTAUSTA E, OJALA R, KIVIRANTA
I, HÄKKINEN A, HEINONEN A
Centre for Arctic Medicine at Thule institute, P.O. Box 5000, 90014 University of Oulu, Finland

The aim of the study was to evaluate reproducibility of the dGEMRIC (delayed Gadolinium
Enhanced Magnetic Resonance Imaging of Cartilage) method in knee cartilage between I and II
measurements compared to II and III with ten asymptomatic adults. The dGEMRIC experiment was
repeated for tibiofemoral and patellar cartilages three times with an average interval of five days
between scans. The measurements were performed by four technicians, whose turns of making the
measurements were coincidental during the study. At first, the participants went through clinical
MRI-series. Then a Gadolinium-oriented contrast agent was intravenously injected. After 90 minutes
penetration time, the T1 weighted GEMRIC imaging was made according the clinical series`
landmarks. Particular ROI (region of interest) segments and their reproducibility were separately
analysed and computed from femoral, tibial and patellar cartilages. Also the bulk values for entire
cartilage were computed. The reproducibility between the measurements were analysed on the group


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level by Coefficient of Variation (CVrms) and Intraclass Correlation Coefficient (ICC). The
reproducibility was moderate or good between the I and II measurements (CVrms 4.1- 10.2 %, ICC
0.47 to 0.98) and between the II and III measurements (CVrms 3.3–13.6 %, ICC 0.47 to 0.99) in all
ROI segments. The reproducibility was slightly lower in the bulk values both between the I and II
measurements (CVrms 9.9– 16.6 %, ICC 0.86 to 0.95) and the II and III measurements (CVrms 9.8
– 17.8 %, ICC 0.88 to 0.98). According to the results, the reproducibility of dGEMRIC method can
be mainly considered good in the segments of femoral, tibial and patellar cartilages both in deep and
in superficial layer regardless of the variation of technicians.

P35
Collagen XIII is a postsynaptic, multifunctional neuromuscular junction protein
LATVANLEHTO A, FOX MA, OIKARAINEN T, TU H, SORMUNEN R, KOSKI A, KALLIO M,
SANES JR, PIHLAJANIEMI T
Institute of Biomedicine, Division of Medical Biochemistry and Molecular Biology, Biocenter Oulu,
P.O. Box 5000, 90014 University of Oulu, Finland

Objectives; We have generated two targeted mouse lines; 1) a LacZ reporter line to study the exact
location and 2) a null line lacking any collagen XIII expression to study biological function of the
transmembrane collagen XIII, also found as a shed, soluble protein. Methods and results; Beta-
galactosidase marker stainings showed that collagen XIII is highly expressed postsynaptically at the
neuromuscular junction (NMJ). The endplate structure in homozygous mice in both lines lacking
intact collagen XIII was smaller and more fragmented than that seen in controls. These structural
changes were accompanied by an electrophysiologically measurable defect in nerve conduction
study. Structural and functional defects of the NMJ had impact on mouse weight, behaviour and
general condition. Mice developed myopathy at old age, but yet the structure of NMJs was more
disturbed in young than in old mice. It was found that postnatal maturation of the postsynaptic
apparatus was delayed and the alignment of pre- and postsynaptic portions was incomplete at some
junctions. Recombinant extracellular fragment of collagen XIII had in vitro a minor effect on
presynaptic differentiation of motoneurons, but it speeded up the postsynaptic maturation on
cultured myotubes. Conclusions; These studies thus indicate that collagen XIII contributes to
stabilization of the NMJ structure and promotes postsynaptic maturation, and this may occur in an
autocrine fashion.

P36
Effects of dexamethasone and exercise on rat myocardial collagen synthesis
AHTIKOSKI AM, TAKALA TES
Department of Health Sciences, University of Oulu, Department of Sports Medicine, Oulu
Deaconess Institute, City of Rovaniemi, Finland

AIM: The aim of this study was to find out effects of dexamethasone treatment and exercise on
collagen expression in rat cardiac left ventricular wall. METHODS: Collagen synthesis was
measured as prolyl 4-hydroxylase activity and type I and III collagen mRNA levels. Collagen
degradation was studied by measuring mRNA level and quantity of gelatinases. Rats were treated
daily with dexamethasone or saline for 3 or 10 days. Eight groups of dexamethasone- treated and
control rats were engaged to daily endurance or sprint running exercise. Total duration of endurance
running was 60 min within 3 days, and 5 hours 15 min within 10 days. Sprint running consisted of

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progressively repeated (5-15 times) 30 s spurts to a mild uphill (5°). RESULTS: Three-day
dexamethasone treatment decreased type I and III collagen mRNA levels. Ten-day dexamethasone
treatment decreased also the activity of prolyl 4- hydroxylase and the quantity of pro-matrix
metalloproteinase-2. Exercise by itself did not affect collagen synthesis, but in combination with
dexamethasone treatment sprint running prevented most of the dexamethasone-induced reduction in
collagen synthesis. CONCLUSIONS: Dexamethasone treatment reduced the synthesis and probably
also the degradation of collagens in the myocardium. Novel finding was also that sprint running was
clearly more effective than endurance running in prevention of these glucocorticoid-induced
changes. The high loading of the heart during sprint running possibly requires preservation of the
extracellular matrix, which distributes the forces generated by myocardial contractions.

P37
Time-course of exercise needed for bone changes in a 12-month intervention study
HEIKKINEN R, KORPELAINEN R, VAINIONPÄÄ A, LEPPÄLUOTO J, JÄMSÄ T
Department of Medical Technology, Institute of Biomedicine, P.O.BOX 5000, 90014 University of
Oulu, Oulu, Finland

The aim was to analyze daily physical activity during a 12-month exercise trial to determine the
duration of regular exercise needed to obtain positive bone changes in previously sedentary women.
Physical activity was continuously assessed with a waist-worn accelerometer in 34 premenopausal
women (35–40 years) participating in an exercise intervention (Vainionpää et al. 2005) with
progressive high-impact training three times a week. The average daily numbers of impacts for each
subject were calculated at five acceleration levels (from 0.3g to 9.2g, see Vainionpää et al. 2006)
during time intervals of 0–3, 0–6, and 0–12 months. Impact activity data were correlated with 12-
month bone density and geometry changes measured by dual x-ray absorptiometry, quantitative
ultrasound and quantitative computed tomography. Femoral neck bone mineral density (BMD)
changes were significantly correlated with impact activity at six and twelve months at high intensity
levels (>3.9g). Trochanter BMD changes were associated with exercise even during the first three
months at impacts exceeding 1.1g. Similarly, mid-femur bone geometry and calcaneal ultrasound
changes were associated with impact activity within the three months of exercise. Six months of
high-impact activity training were positively associated with bone changes at the femoral neck,
while a shorter period was related with changes in trochanter, mid-femur and calcaneus. The results
can be utilized in the design of training programs to prevent bone loss in premenopausal women.
References: Vainionpää, A., Korpelainen, R., Leppäluoto, J. & Jämsä, T. 2005. Osteopor Internat 16,
191-197. Vainionpää, A., Korpelainen, R., Vihriälä, E., Rinta-Paavola, A., Leppäluoto, J. & Jämsä,
T. 2006. Osteopor Internat 17, 455-463.

P38
Impact exercise alters long-term bone turnover in a dose-dependent manner
VAINIONPÄÄ A, KORPELAINEN R, VÄÄNÄNEN HK, HAAPALAHTI J, JÄMSÄ T,
LEPPÄLUOTO J
Department of Medical Technology and Department of Physiology, Institute of Biomedicine,
University of Oulu, P.O. Box 5000, 90014 Oulu, Finland

Impact exercise increases bone mineral density and improves bone geometry in weight-bearing
bones. However, long-term effects of impact exercise on bone turnover as well as dose-dependency

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are not known. To study the effects of high-impact exercise on bone turnover and calciotropic
hormones, we performed a 12-month population-based, randomized, controlled exercise trial in 120
women (age 35-40 years). Subjects were randomly assigned to an exercise group (EG; n=60) or a
control group (CG; n=60). The exercise regimen consisted of supervised high-impact exercises three
times per week. Daily impact loading was assessed by using an accelerometer (Newtest Ltd.,
Finland). Bone turnover markers, calciotropic hormones and turnover balance (Uncoupling Index,
UI=Z(PINP)–Z(TRACP5b)) were analyzed at 0, 6 and 12 months. Thirty-seven women in the EG
and 39 women in the CG completed the study. The Uncoupling Index increased in the EG and
decreased in the CG (0.32 vs. -0.28; p=0.03). PTH decreased significantly more in the EG than in
the CG (-11.2 vs. -2.2 pg/mL; p=0.03). The most active subjects with the highest number of impacts
exceeding accelerations of 2.5g had higher increases in the UI compared to the least active subjects
(0.86 vs. -0.48; p≤0.001). The number of these impacts separating the most and the least active
subgroups was 400 impacts per day equivalent to half an hour of impact exercise like jogging,
running or jumping. In conclusion, regular impact exercise alters bone turnover balance in favour of
bone formation in a dose-dependent manner. Effective dose is reached during normal exercise
training indicating potential to affect bone strength with impact exercise. The Uncoupling Index
seems to have potential to reflect the effect of exercise on bone turnover.

P39
Contractile defects and protein nitration in slow-twitch muscle of mice with collagen-induced
arthritis
YAMADA T, ZHANG SJ, BRUTON JD, WESTERBLAD H
Dept of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden

In patients with rheumatoid arthritis (RA), there is a progressive muscle weakness together with joint
dysfunction. However, there is no data as to whether the muscle fibres‟ intrinsic contractile
properties are affected in RA. We investigated the muscle contractility in slow- twitch soleus and
fast-twitch extensor digitorum longus (EDL) muscle of mice with collagen- induced arthritis (CIA).
The results show a markedly decreased force per cross-sectional area in soleus muscles but not in
EDL muscles of CIA mice compared to control mice. To assess the underlying mechanism of this
force deficit, we focused on the metabolism of reactive oxygen and nitrogen species. The results
showed a marked increase in protein nitration in CIA soleus muscle and the extent of protein
nitration correlated with the force decrease. In CIA soleus muscle there was a significant increase in
neuronal nitric oxide synthase (NOS) expression but not in inducible or endothelial NOS expression.
There was no difference in superoxide dismutase 1 or 2 expression between CIA and control soleus
muscles. In conclusion, our results provide the first evidence that arthritis can cause decreased force
production in slow-twitch muscle and that protein nitration appears to have a central role in this
impairment.




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                          Poster session 07 – Gastrointestinal physiology

P40
Oral bacteria regulate gastric mucosal defense via bioactivation of dietary nitrate
PETERSSON J, SCHREIBER O, PHILLIPSON M, ROOS S, JANSSON EÅ, LUNDBERG J,
HOLM L
Dept. of Medical Cell Biology, Uppsala University, Uppsala, Sweden

Background and Aims: Dietary nitrate enhances gastric mucosal defense by luminal NO production,
as oral bacteria reduce salivary nitrate to nitrite, which is further reduced to bioactive nitric oxide in
the acidic stomach. The bacterial conversion of nitrate to nitrite is a crucial step in the formation of
the protective NO in the stomach. We evaluated the importance of oral bacteria during nitrate
supplementation on gastric mucosal defense in rats and mice. Methods: Regular drinking water or
nitrate supplemented water was given to the animals for one week. The oral bacterial flora was
suppressed with an antiseptic mouth spray twice daily during this period. Intragastric NO and plasma
nitrite were measured by chemiluminescence. The thickness of the gastric mucus layers was
measured in vivo with the animal´s stomach exteriorized for intravital microscopy. Additionally, the
stomach was challenged with diclofenac and the gastric levels of P-selectin were quantified using the
dual labeled antibody technique. Results: Nitrate supplementation resulted in increased levels of
intragastric NO and plasma nitrite. These effects were strongly reduced in animals with a suppressed
oral flora. Nitrate supplementation increased the thickness of the adherent gastric mucus layer. This
protective effect was totally absent in mouth-sprayed animals. Dietary nitrate protected against
diclofenac induced damage and this protection was abolished if the oral flora was reduced.
Conclusions: The formation of intragastric NO from dietary nitrate is critically dependent on oral
bacteria. Without these bacteria, the gastroprotective role of nitrate is abolished.

P41
Influence of short glyprolines with arginine on C-end on gastric mucosa homeostasis in
ethanol-induced ulcerogenesis
PUCHKOVA AN
Dept. Human & Animal Physiology, Biology Faculty, Lomonosov Moscow State University,
Leninskie Gori 1, build. 12, 119991, Russia

The aim of the study is to evaluate the ability of short glyprolines with arginine on C-end - PGPR,
PGR and GPR – to increase gastric mucosa resistance to damaging factors, such as ethanol. The
reason for the study is that glyprolines effectively support disrupted gastric mucosa homeostasis,
among their other effects. PGP and PG demonstrated different combinations of protective effects on
different gastric ulcer models (stress- and ethanol-induced ulcers, acetate ulcer). According to recent
data, as GPR has already shown biological activity not connected with gastric mucosa homeostasis:
it successfully inhibited ADP-induced platelet aggregation and prevented neuronal death caused by
beta-amyloid. So it is perspective to study influence of glyprolines on C-end on gastric mucosa
homeostasis. In this work we investigated their possible effect on ethanol ulcer model. Peptides were
administered intragastrically in 3.7 µmol/kg dose 1 hour prior to ethanol. 5 ml/kg dose of 96%
ethanol was used to induce gastric mucosa damage. PGPR and PGR showed significant decrease in
size of mucosa lesions (32.47±12.39% and 41.78±25.53% of control size respectively). GPR was
ineffective on ethanol model (93.37±81.54% of control size). Glyprolines with arginine on C-end


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                                             Abstracts

showed similar efficacy as original glyprolines: PGP and PGPR antiulcer effects are almost the same
(64.0% and 67.53% respectively) and PGR was less effective than PG (58.22% compared to 83.1%).
Conclusions: besides already known biological activities of glyprolines with arginine on C-end,
peptides PGPR and PGR demonstrate significant protective antiulcer effect on ethanol-induced ulcer
and GPR is ineffective.

P42
Peripheral and central mechanisms of gastric mucosa homeostasis and glyprolines
SAMONINA GE, KOPYLOVA GN, UMAROVA BA, BARAEVA ZV, BADMAEVA KE,
TRUFANOVA AV
Dept. Human & Animal Physiology, Biology Faculty, Moscow State University, Leninskie Gori 1,
build.12, Moscow, 119991, Russia

Glyprolines are the new family of regulatory peptides consisting of glycine and proline [Ashmarin et
al., 2002]. Influence of glyprolines beginning from di- to hexaglyprolines (3.7mkmol/kg, per os) on
gastric mucosa (GM) homeostasis (H) was studied using ethanol (E), stress (S) and acetate (A)-
induced ulcer in male rats. A possibility not only to increase GM stability to E and S but also to
accelerate A ulcer healing was shown. All studied glyprolines accelerated A ulcer healing but not all
of them increased GM stability to E or S. We supposed that it is connected with vague influence of
glyprolines on different peripheral or central mechanisms of ulcerogenesis. As it‟s known,
pathogenesis of E ulcer is mostly determined by peripheral mechanisms and S – by central ones.
Only two peptides – Pro-Gly-Pro (PGP) and Gly-Pro-Gly-Pro- Gly-Pro - revealed both protective
and therapeutic effects. PGP affects different peripheral and central ulcerogenesis mechanisms. PGP
decreases or prevents post-stress disorders of behavior, mesentery microcirculation, mast cells
degranulation, testifying its influence on the CNS integrating functions. This glyproline could also
exhibit direct protective effects on the cell element of GM itself and also on neural structures
involved in the maintenance of GM H: PGP prevents ischemia in gastric tissue induced by
indomethacin or E administration, induced a direct vasorelaxant effect, increased the contractive
activity of mesenteric lymphatic microvessels, improved rheological properties of blood, decreased
vagal-stimulated acid secretion and mast cells degranulation in vitro. So, some glyprolines display
protective and curative gastric mucosa antiulcer properties, recovering affected central and
peripheral mechanisms of GM H.

P43
GFRα2 knockout mouse as a model for studying autonomic regulation of gastric acid and
ghrelin secretion
KUPARI J, ROSSI J, HUOTARI A, VOIKAR V, HERZIG K-H, AIRAKSINEN MS
University of Helsinki, Neuroscience Center, Viikinkaari 4 Cultivator II, 00014 Helsingin yliopisto,
Finland

GDNF-family receptor α2 (GFRα2) and its ligand neurturin are essential for cholinergic target
innervation in parasympathetic and a subset of myenteric neurons. Here, we observed a deficit of 80-
90% of VIP and VAChT positive nerve fibers and associated glial cells in stomach mucosa in mice
lacking GFRα2 (KO). Consistent with this, neurturin mRNA is expressed specifically in the basal
part of the gastric mucosa in postnatal and adult mice. No difference in basal gastric pH (WT
2.2±0.37, KO 2.3±0.25) or gastric acid content (WT 0.24±0.02 μmol H+/g, KO 0.25±0.03 μmol

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                                            Abstracts

H+/g) was seen between the phenotypes. However, vagal stimulation with 2-DG had little effect on
KO acid secretion (~2 fold increase) when compared to WT animals (~8 fold increase). Ghrelin is an
orexigenic peptide hormone produced primarily in the basal part of stomach mucosa. Cholinergic
innervation has been proposed to regulate ghrelin secretion but factors promoting innervation of
ghrelin producing cells are poorly known. Plasma ghrelin levels in KO animals were ~40% higher
when compared to WT. However, no difference in mRNA levels of ghrelin between the genotypes
was seen, suggesting increased ghrelin secretion in KO animals. KO mice showed abnormal feeding
behaviour: shorter meal intervals and longer total feeding time. After overnight fast, the time and
number of meals needed to reach the first long meal interval were increased in KO mice, suggesting
reduced satiety. Unexpectedly, the time needed to initiate feeding after food re-presentation was
longer in KO animals. Taken together, our results indicate that cholinergic innervation of gastric
mucosa requires GFRα2 signaling and that GFRα2-KO mice provide a useful model for studying
autonomic neuronal regulation of gastric acid and ghrelin secretion.

P44
Defective jejunal and colonic salt absorption and altered Na+/H+ exchanger 3 (NHE3) activity
in NHE regulatory factor 1 (NHERF1) adaptor protein deficient mice
BROERE N, CHEN M, CINAR A, SINGH AK, HILLESHEIM J, BIEDERER B, LÜNNEMANN
M, ROTTINGHAUS I, KRABBENHÖFT A, ENGELHARDT R, RAUSCH B, WEINMAN EJ,
DONOWITZ M, HUBBARD A, KOCHER O, DE JONGE HR, HOGEMA BM, SEIDLER U
Gastroenterology, hepatology and endocrinology, Medical School Hannover, 30625, Hannover,
Germany

We investigated the role of the Na+/H+ exchanger 3 (NHE3) regulatory factor 1 (NHERF1) on
intestinal salt and water absorption, brush border membrane morphology and on the NHE3 mRNA
expression, protein abundance, and transport activity in the murine intestine. NHERF1-deficient
mice displayed reduced jejunal fluid absorption in vivo, as well as a attenuated Na+ absorption in
isolated mucosa of jejunal and colonic tissue but not of ileal mucosa in vitro. However, cAMP-
mediated inhibition of both parameters remained intact. The basal NHE3 transport rate was
decreased in surface colonocytes, while inhibition by cAMP and cGMP was normal.
Immunodetection of NHE3 revealed normal NHE3 localization in the brush border membrane
(BBM) of NHERF1 null mice, but NHE3 abundance, as measured by Western blot, was significantly
reduced in isolated membranes from the small and large intestine. Furthermore, the microvilli in the
proximal colon, but not in the small intestine, were significantly shorter in NHERF1 null mice.
Additional knockout of PDZK1 (NHERF3), another member of the NHERF family of adapter
proteins, which binds to both NHE3 and NHERF1, further reduced basal NHE3 activity and caused
complete loss of cAMP-mediated NHE3 inhibition. An activator of the exchange protein activated
by cAMP (EPAC) had no effect on jejunal fluid absorption in vivo, but slightly inhibited NHE3
activity in surface colonocytes in vitro. In conclusion, NHERF1 has segment-specific effects on
intestinal salt absorption, NHE3 transport rates and NHE3 membrane abundance without affecting
mRNA levels. However, unlike PDZK1, NHERF1 is not required for NHE3 regulation by cyclic
nucleotides.




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                                             Abstracts

P45
Dual role of the Na+/H+ exchanger isoform 3 for PEPT1-mediated H+/dipeptide cotransport in
native murine intestine
CHEN M, SINGH A, DRINGENBERG U, YERUVA S, WANG J, ENGELHARDT R, RIEDERER
B, MANNS MP, RUBIO-ALIAGA I, NÄßL A, SOLEIMANI M, SHULL G, DANIEL H,
SEIDLER U
Gasteroenterology, Hepatology and Endocrinology, Medical School Hannover, 30625, Hannover,
Germany

Backround: Inhibition of the apical Na+/H+ exchanger isoform 3 (NHE3) interferes with dipeptide
uptake in intestinal cell lines. Aim: This study investigates the mode of coupling between electrolyte
and nutrient transporters involved in H+-symport driven dipeptide uptake in native murine intestine.
Methods and Results: The luminal application of the dipeptide Gly-Sar resulted in a strong increase
in murine small intestinal fluid absorption and a decrease in villous enterocyte pHi, measured by
two-photon microscopy in vivo. It caused a strong short circuit current (Isc) response in chambered
jejunal mucosa in vitro, and a decrease in pHi in BCECF-loaded enterocytes in isolated jejunal villi.
Genetic ablation of PEPT1 abolished Gly- Sar-induced fluid absorption, Isc response, and enterocyte
acidification. Genetic ablation of NHE3 did not change the distribution or amount of PEPT1 in the
brush border membrane, but abolished Gly-Sar-induced fluid absorption, strongly diminished Isc
response, but significantly enhanced enterocyte acidification in the absence but not the presence of
CO2/ HCO3- . Genetic ablation of the apical Cl-/HCO3- exchanger Slc26a6 did not influence Gly-Sar-
induced Isc, but significantly enhanced enterocyte acidification in the presence but not the absence
of CO2/HCO3-. The transmembrane Na+ was more important than the proton gradient to sustain high
rates of H+/dipeptide transport in the native epithelium. Conclusions: NHE3 and Slc26a6 are equally
important for recovery of Gly- Sar-induced cellular acid loads, but only NHE3 is essential for
H+/dipeptide transport. Via H+ recycling through NHE3, the transmembrane Na+ gradient energizes
H+/dipeptide transport in the native epithelium even in the complete absence of a proton gradient.

P46
Differential role for the PDZ proteins NHERF1, NHERF2 and PDZK1 in the regulation of
CFTR-mediated intestinal anion secretion in vivo
SINGH AK, RIEDERER B, KRABBENHÖFT A, RAUSCH B, DE JONGE HR, DONOWITZ M,
WEINMAN EJ, KOCHER O, HOGEMA BM, SEIDLER U
Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School,
Hannover 30625, Germany

Heterologous expression studies have demonstrated that the PDZ-proteins NHERF1, NHERF2 and
PDZK1 (NHERF3) modulate CFTR membrane expression, conductivity and interaction with other
proteins. To study their biological roles in vivo in an epithelium that highly expresses both CFTR
and all three NHERF proteins, we investigated the effect of NHERF1, 2 and PDZK1 ablation, or a
combination of the above, on duodenal HCO3- secretion in the basal state and after agonist and
inhibitor application. The proximal duodenum of anesthetized mice was perfused in situ, and HCO3-
secretion was determined by back-titration. NHERF1 ablation strongly reduced basal and forskolin
(FSK)-stimulated HCO3- secretory rates, and completely prevented 2-adrenergic stimulation.
NHERF2 deletion significantly augmented FSK-stimulated HCO3- secretion, prevented the
inhibitory effect of LPA, and partially rescued the suppressed basal HCO3- secretion resulting from

                                                 122
                                              Abstracts

NHERF1 ablation. PDZK1 ablation reduced basal HCO3- secretion but not the response to FSK. The
deletion of CFTR abolished agonist-mediated HCO3- secretion and any effect of NHERF ablation.
We conclude that the three NHERF proteins differentially modulate basal and agonist-mediated
duodenal HCO3- secretion in vivo in a CFTR- dependent fashion. NHERF1 is an obligatory linker for
2-adrenergic stimulation of CFTR, and strongly augments cAMP-mediated stimulation. NHERF2
confers inhibitory signals, i.e. as a coupling factor between inhibitory LPA receptors and CFTR.

P47
Key role of CFTR for all modes of intestinal HCO3- secretion
SINGH AK, ZHENH W, SJOBLOM M, SEIDLER U
Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School,
Hannover 30625, Germany

Background: CF patients suffer from a variety of gastrointestinal problems, which may all be
directly or indirectly linked to the inability of the intestinal mucosa to secrete HCO3- and to inhibit
Na+/H+ mediated fluid absorption and proton secretion. This has led to an intense search for
alternative modes of intestinal anion transport, and expressions of a variety of potential alternative
intestinal anion transporters have been reported. Aim: To delineate the dependency of different
modes of intestinal HCO3- secretion on CFTR expression. Methods and Results: We studied acid-,
agonist-, and HCO3- stimulated, as well as Cl--dependent HCO3- secretion in the CFTRtm1cam and
WT murine duodenum in vivo. NHE3 and Slc26a6-deficient mice were used for selected questions.
Luminal acid, forskolin, heat-stable E. coli enterotoxin (STa), PGE2, carbachol, and an increase of
blood HCO3- all stimulated duodenal HCO3- secretion in anesthetized WT but not in CFTRtm1cam
mice. Pharmacological inhibition or genetic ablation of NHE3 resulted in a significantly higher basal
HCO3- secretory rate, which was electroneutral and therefore due to an unmasking of apical Cl-/
HCO3- exchange activity. Accordingly, Slc26a6 ablation attenuated S1611- induced J HCO3-
Removal of luminal Cl- reverted basal HCO3- secretion to H+ secretion, but surprisingly, forskolin
was able to elicit a full HCO3- secretory response. In the absence of CFTR, electroneutral NaCl
absorptive rates were similar to WT rates, but S1611 induced virtually no increase in HCO3-
secretion. Conclusion: This indicates that the apical anion exchangers Slc26a6 and Slc26a3 need
proton recycling via NHE3 to operate in the Cl- absorptive mode, and Cl- exit via CFTR to operate in
the HCO3- secretory mode. In addition, Cl- independent HCO3- secretion can be stimulated by cAMP
increase, likely from the crypt region, which expresses high levels of CFTR but none of the anion
exchangers.

P48
Chronic PD induces remodeling of the peritoneal membrane and local secretion of TGF- and
VEGF in rats
OLSEN SJ
Jonas Liesv 99, 5009 Bergen, Norway

In peritoneal dialysis (PD), the frequent exposure to dialysis fluids elicits a chronic state of a low-
grade peritoneal inflammation. On the morphological level, fibrosis and angiogenesis is common
after chronic PD, as are abnormalities in the peritoneal membrane. In the long run, the dialysis
capacity of the peritoneal membrane is lost due to these changes. Under isoflurane anesthesia,
injection ports were implanted and attached catheters were tunneled to the peritoneal cavity. Treated

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                                             Abstracts

group (PD; n=8) received PD twice daily, while catheter control group (CC; n=7) just had catheters
and ports implanted. Negative controls received no treatment (NC; n=5). After 8 weeks, interstitial
fluid (IF) from peritoneum was isolated using a centrifugation method and analyzed for cytokine
content along with plasma. Collagen and hyaluronic acid (HA) was measured in the peritoneal
tissue. The results showed a local production of TGF- in peritoneal tissue in the PD group (p<0.05).
VEGF was not changed due to PD. Collagen content in PD (p=0.03) and in CC (p=0.005) was
higher as compared to NC. HA increased in PD (p=0.018), with a trend towards increase in CC
(p=0.072) compared to NC. In conclusion, the local production of TGF- in the peritoneal IF in
response to chronic PD may be an integral component in the development of fibrosis. However,
VEGF levels were generally low, and other mediators of angiogenesis may be involved in the
process. The changes in the collagen and HA contents in CC indicate that the catheter per se may
alter the extracellular matrix composition.




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                                              Abstracts


                               Poster session 08 – Neurophysiology I

P49
Interspike intervals of human motor units during voluntary recruitment
MEIGAL A, TITOVA E
Department of Human and Animal Physiology, Lenin Str., 33, 185910 Petrozavodsk, Russian
Federation

The objective of the present study was to analyze interspike intervals (ISI) of the human motor unit
(MU) discharges in respect with the motor task. The subjects (n=4, 1 m, 3 f, 8 MUs, 2452 ISI) after
preliminary practicing were instructed to voluntarily recruit the ordered number of MU discharges of
m. triceps brach. (from 1 to 7, 40 attempts for each task), with the help of audio- visual feedback
from the EMG device. From 50 to 90% attempts were correct. In these correct attempts in the task “2
discharges” the ISI was 99.45 ms. In the task “3 discharges” the 1st ISI was 100.57 ms, and the 2nd
was longer (127.57 ms, p<0.01). In the tasks “4, 5, 6 and 7 discharges” the ISIs presented a wave-
like sequence: the first three ISIs were increasing in a row, but the 4th was always shorter than the
3rd ISI. The 5th and the 6th ISI were again longer than the 4th ISI. Also, the 1st ISI was decreasing
with the increasing number of discharges in the task (from cal. 100 to 80-85 ms). In conclusion, the
ISI sequence may reflect the code of “motor command”. Further it would be essential to compare the
ISI sequence in the correct attempts to the “mistake attempts” (with 1-2 extra and 1-2 lacking MU
discharges) and also to the “no-task attempts”.

P50
Coenzyme Q10 and alpha lipoic acid supplementation to diabetic rats: Conduction velocity
distributions
AYAZ M, TUNCER S, OKUDAN N, GÖKBEL H
Department of Physiology, Meram Faculty of Medicine, Selçuk University, Konya, Turkey

Objective: Current study aimed to investigate diabetes- and coenzyme Q10 (CoQ10) and alpha lipoic
acid supplementation- induced changes in the conduction velocity distributions of the rat sciatic
nerve fibers. Methods: Sciatic nerve compound action potentials (CAP) were recorded by suction
electrode and the conduction velocity distributions were done by collision technique. Results:
Diabetes resulted in the significant increase in time to peak, rheobase and chronaxie values of these
CAP waveforms. Meanwhile the maximum depolarization, area, kinetics and the conduction
velocities of both fast and slow nerve fiber groups were found to be decreased. CoQ10 was found to
have some positive effects on the diabetes induced alterations. This supplementation induced
positive changes mainly seen in the area and fall down phase of the kinetics of CAP waveforms as
well as rheobase, chronaxie and speed of the intermediately conducting groups (approximately 40
m/sec). Alpha lipoic acid supplementation did not produce any effect. The current study has shown
for the first time that diabetes induced shift of actively contributing nerve fibers towards the slower
conduction velocities. Meanwhile supplementation of CoQ10 not only stopped this shift but also
started to restore towards the age matched control group values. Conclusion: Besides the altered
mitochondrial phenomenon, positive effects seen on diabetic neuropathy can be attributed to
antioxidant property of CoQ10.




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                                              Abstracts

P51
EEG burst suppression recorded from depth electrodes during anesthesia
JÄNTTI V, KAUPPINEN M, PUUMALA P, SONKAJÄRVI E, KARVONEN E, KAUPPINEN P,
OLLILA K, RYTKY S
Department of Physiology, Medical School, University of Tampere, Finland

Objective: Analysis of local activity and cortical activity recorded from electrodes in subthalamic
nucleus. Methods: The patients were operated to implant electrodes in subthalamic nucleus to treat
parkinsonism. Three patients were anesthetized with propofol, three with sevoflurane and three with
desflurane without other simultaneous drugs. EEG was recorded with Neuroscan TM or Nervus ™
digital EEG equipment simultaneously from scalp and depth electrodes. Results: Burst suppression
was recorded between two depth electrodes and between two scalp electrodes. The patterns of bursts
in these two traces resembled each other closely, and suppression occurred synchronously. During
suppression we could see patterns in depth electrodes which were not visible on scalp. Conclusion:
The similarity of burst suppression EEG recorded with scalp and depth electrodes can only be
explained with mainly cortical origin of the signal. During suppression we can see transients and
rhythms in depth electrodes which are not seen on scalp and we conclude that these are so called
local field potentials. Mathematical modeling of the intracerebral electrical fields shows why cortical
activity is recorded with high amplitude between two depth electrodes. Intracerebral electrical
currents are, however, very complex due to, among other things, anisotropy of subcortical brain.
Improved models of these fields are necessary to understand the signals recorded with subcortical
electrodes, the effect of fields created by stimulation currents in these electrodes, as well as in
understanding the subcortical components of evoked potentials.

P52
TIRF study of P2X3 receptors trafficking in cultured neurons
PRYAZHNIKOV E, GINIATULLIN R, KHIROUG L
Viikinkaari 4, Neuroscience Center, University of Helsinki, Helsinki, Finland

Physiological studies using P2X receptor agonists and antagonists have suggested that activation of
P2X3 receptors by extracellular ATP depolarizes nociceptive neurons to evoke a sensation of pain
(Bland-Ward and Humphrey, 2000). In order to trigger pain in response to ATP, purinergic receptors
must reside in plasma membrane. Receptors insertion in plasma membrane is a dynamic process
which is counterbalanced by receptor internalization by endocytotic mechanisms. Constant recycling
of membrane- inserted fraction of P2X receptors has been suggested to be affected in neuropathic
pain (Xu and Huang, 2004) and therefore represents an attractive target for pain treatment. In our
work we studied trafficking of fluorescently tagged P2X3 receptors in cultured hippocampal neurons
using Total Internal Reflection Fluorescence microscopy (TIRF). TIRF is an advanced optical
microscopy method based on unique properties of evanescent field that is created on the surface of
cell-bearing glass due to total internal reflection of the laser beam. Evanescent field allows selective
visualization of those fluorescent molecules which are located in the immediate vicinity (~100 nm)
of the plasma membrane of cultured cells (Pryazhnikov and Khiroug, 2007). In our study we
selectively tracked perimembrane pool of P2X3 receptors which has greater probability of being
inserted in the plasma membrane. We found agonist induced reduction of membrane associated pool
of P2X3 receptors which is consistent with internalization process and may regulate the response of
neurons to ATP. Bland-Ward PA, Humphrey PP. 2000. J Auton Nerv Syst 81:146-151. Pryazhnikov

                                                  126
                                             Abstracts

E., Khiroug L. 2008. Glia. 56(1):38- 49. Xu GY, and Huang LY. 2004. Proc Natl Acad Sci U S A.
101(32):11868-73.

P53
Interactions with calnexin and ER calcium pump SERCA2b regulate human delta opioid
receptor maturation
TUUSA JT, LESKELÄ TT, PETÄJÄ-REPO UE
University of Oulu, Institute of Biomedicine, Department of Anatomy and Cell Biology, P.O.Box
5000, 90014 Oulu, Finland

The human delta opioid receptor (hOR) was used as a model to study the role of ER calcium pump,
Sarco(endo)plasmic reticulum ATPase 2b (SERCA2b), in the biogenesis of G protein-coupled
receptors (GPCRs). Stable cell lines with inducible expression of the hOR and wild-type or mutant
SERCA2b with various tags were created. Transient transfections were used for co-expression,
interactions were examined by co-immunoprecipitation of native or chemically crosslinked proteins,
receptor maturation was followed by pulse-chase assay and Fluo-3/AM was used to measure ER-
released calcium by confocal microscopy. We have earlier reported that newly synthesized hOR
precursors interact with SERCA2b and also with ER molecular chaperone calnexin. Calnexin has
also been suggested to modulate SERCA2b activity in vivo. Here we show that SERCA2b, calnexin
and the receptor precursor form a ternary complex that is not dependent on receptor N-glycans.
Different domains of SERCA2b mediate the interaction with calnexin and the receptor. Also,
calcium and ATP had a different modulatory effect on the interactions. The co-expression of
catalytically inactive SERCA2b mutant (D351A) reduced receptor maturation without a collapse in
ER calcium level. The decrease in hOR maturation was similar to the effect of SERCA inhibitor
thapsigargin (Tg). However, unlike Tg, mutation of the active site of SERCA2b, which stabilizes a
different conformation of the calcium pump, did not abolish receptor interaction. We conclude that
the active SERCA2b modulates GPCR biogenesis via dynamic protein-protein interactions.

P54
Taurine reduces ethanol-induced caspase-3 activation in the developing cerebellum
TARANUKHIN A1,2, TARANUKHINA E1, SARANSAARI P1, DJATCHKOVA I1, PELTO-
HUIKKO M1,3, OJA SS1
1
  Medical School, University of Tampere, Finland, 2Laboratory of Comparative Somnology and
Neuroendocrinology, Sechenov Institute of Evolutionary Physiology and Biochemistry,
St.Petersburg, Russia, 3Department of Pathology, University of Tampere, Finland

Taurine is a sulphur-containing amino acid with multiple functions, including cell protection. In the
present work we studied the possible neuroprotective effect of taurine on ethanol-induced apoptosis
in 7-day-old male mice whose cerebella are extremely sensitive to ethanol during synaptogenesis.
The degree of apoptosis was estimated with immunostaining for activated caspase-3 in mid-sagittal
paraffin-embedded sections containing lobules II-X of the cerebellum. Eight hours after ethanol
administration (total dose 5 g/kg) the number of activated caspase-3-immunoreactive neurons was
increased in the external and internal granular layers of all cerebellar lobules, indicating ethanol-
triggered apoptotic neurodegeneration. Taurine treatment (two injections per 1 g/kg each)
significantly decreased the number of activated caspase-3-immunoreactive cells in both external and
internal granular layers after ethanol administration but its effect varied in different lobules. We

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                                             Abstracts

conclude that taurine protects immature cerebellar neurons, reducing markedly ethanol-induced
caspase-3 activation but does not abolish it completely. This study was supported by the Center for
International Mobility (grants EH-04-2615 and MK-05-3550), the competitive research funding of
Pirkanmaa Hospital District (grants 9F070, 9G051, 9G203) and the Finnish Cultural Foundation
(2007).

P55
Differential roles of L-type Ca2+ channels for evoked and spontaneous transmitter release onto
rat preoptic neurons
MALININA E, DRUZIN M, JOHANSSON S
Section for Physiology, Department of Integrative Medical Biology, Umeå University, 901 87 Umeå,
Sweden

L-type Ca2+ channels are thought not to be critical for synaptic transmitter release evoked by low-
frequency stimulation, but do in some cases contribute to post-tetanic potentiation (PTP) of evoked
release. We have previously shown that L-type channels are present in presynaptic terminals on
medial preoptic neurons from rat and here analyse their roles in evoked as well as spontaneous
transmission, by perforated-patch recording in brain slices. At low-frequency (0.5 – 2.0 Hz)
presynaptic stimulation, the L-type channel blocker calciseptine (100 µM) significantly enhanced
GABAA-receptor- mediated as well as AMPA-receptor-mediated postsynaptic currents, by 31±7%
(n= 6) and 18 ±5 % (n = 9) respectively (mean ±S.E.M.). In contrast, the steady frequency of
spontaneous GABA-mediated postsynaptic currents (sIPSCs) was not affected by calciseptine. After
a 10-s period of high- frequency (50 Hz) stimulation, there was PTP of evoked GABAA-receptor-
mediated postsynaptic currents, but they were only slightly affected by calciseptine. The high-
frequency stimulation also induced PTP of the sIPSC frequency. However, after such potentiation,
calciseptine significantly reduced the sIPSC frequency, by 24±10 % (n = 13). Thus, in MPN
neurons, L-type Ca 2+ channels may play a role at low- frequency-stimulation and differentially
affect evoked and spontaneous transmitter release. We speculate that, at low frequency stimulation,
the limited Ca2+ entry through L-type channels affects transmission via Ca2+-gated K+ channels and
effects on the membrane potential, whereas at high-frequency stimulation, a more massive Ca2+
entry induces more direct potentiating effects on the release machinery.

P56
Postponed effects of chronical neonatal arginine-vasopressin structural analogue (Ас-D-Met-
Pro-Arg-Gly-NH2) administration on learning processes in white rats
KIM PA, VOSKRESENSKAYA OG, KAMENSKY AA
MV Lomonosov Moscow State University, 1-12 Leninskie gory, Human and Animal Physiology
Department, Faculty of Biology, Moscow, 119991, Russia

Arginine-vasopressin (AVP) can take not only its essential hormonal effects, but can also provide
various extrahormonal influence. AVP acts not only as a neuropeptide in the adult brain but may
play a significant role during maturation of the central nervous system. The half-life of AVP-
molecule average 10 – 15 minutes. To increase its bioavailability the AVP structural analogue
Acetyl-D-Methionine-Proline-Arginine-Glycine-Amide (Ас-D-Met-Pro-Arg-Gly-NН2) was
synthesized. Previous researches have shown that both AVP and its analogue administration provide
modulatory effects on white rats behavioral skills formation. The presented research is aimed to

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                                             Abstracts

reveal postponed effects of Ас-D-Met-Pro-Arg-Gly-NН2 neonatal administration. Peptide was
administered intranasally in the dose of 10 ng/kg on days 8 to 14 following the day of birth. Control
animals received distilled water. The behavioral reactions of mature animals (63–70 days) were
studied in next tests: “active avoidance task ” and “intricate food maze”. In the “active avoidance
task ” test the increase of number of correct trials on the 2th and on the 4th days of learning and
during the control of the skill conservation was revealed. In the “intricate food maze” test the
increase of the same parameter on the 2nd day of learning, the decrease of exit from the first maze
compartment latency during all period of learning and the decrease of reaction time on the 4th day of
learning were displayed. Data obtained allow to affirm, that neonatal administration of Ас-D-Met-
Pro-Arg-Gly-NН2 lead in mature rats to the improvement of learning with both negative and positive
reinforcement. The possible explanation is that the time of peptide administration is all-important to
vasopressinergic system formation.

P57
Exogenous heat shock protein 70 affects sleep and thermoregulation after sleep deprivation in
pigeons
LAPSHINA KV
Sechenov Institute of Evolutionary Physiology and Biochemistry RAS, Laboratory of comparative
thermophysiology, pr. M. Toreza 44, 194223, St. Petersburg, Russia

The heat shock protein 70 kDa (Hsp70) is known as the molecular chaperone taking a part in the
protein folding and regulation of cell functions. Also it was shown that Hsp70 can regulate
physiological functions at whole organism level (Pastukhov et al. 2004). The central microinjections
of Hsp70 could increase the total time of non-rapid-eye-movement sleep (non-REMS) and decrease
the brain temperature. It is unclear what effect can Hsp70 exert on sleep and thermoregulatory
characteristics during the recovery after sleep deprivation (SD) in pigeons. The total SD was evoked
by a tactile and sound stimulation during 5 hours. Exogenous Hsp70 (purified from contamination
and consisting of constitutive and inducible isoforms) was injected into the 3rd brain ventricle in
dose of 1.5 µg/µl in the end of SD. In control conditions pigeons were injected with vehicle in the
same volume. Recording of polysomnogram and thermoregulatory characteristics during 24 h was
carried out by computer system. The procedure of SD was characterized by an increase in brain
temperature and contractile muscular activity. During the first hour after the end of SD wakefulness
prevailed and the level of contractile muscular activity remained high. The rebound effect of non-
REMS was observed only in the second hour. The injection of Hsp70 after the end of SD evoked the
non-REMS rebound, which occurred during the first hour. During the next 5 hours the amount of
non-REMS was also increased. Hsp70 also induced a decrease in brain temperature and contractile
muscular activity which occurred immediately after the end of SD. Thus we suppose that early
appearance of non-REMS and decrease in brain temperature and contractile muscular activity can be
an evidence of stress-limiting effect of Hsp70. References: 1.Pastuchov Yu.F., Ekimova I.V., Hudik
K.A. & Guzhova I.V. 2004. Dokl.Acad.Sci 402, 275-278




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                                              Abstracts


                         Poster session 09 – Neurophysiology II - GABA

P58
Millisecond substance exposure on HEK-293 cells expressing several GABA-A receptor
subtypes using patch-clamp in DynaflowTM system
GOUISSEM S, STRÖMBERG J, LUNDGREN P, TAUBE M, JOHANSSON I-M, ISAKSSON M,
BÄCKSTRÖM T
Umeå Neurosteroid Research Center, Umeå University, Dept. of Clin. Sci., Obstetrics &
Gynecology, 5B level 5, SE-901 85 Umeå, Sweden

The GABA-A receptor is in the brain expressed in up to 20 different subunit compositions usually
two  two  and one  subunit. Some of the compositions are dominantly expressed in areas related
to a certain function, like the 5 is located in hippocampus and related to memory while 1 is more
diffusely spread and related to sedation. However, usually there is a mixture of receptor forms in a
certain brain area and therefore a specific receptor subtype is difficult if not impossible to study in
situ. Another problem is that the physiological exposure time of GABA and other compounds is in
millisecond range. To overcome these problems we have developed two HEK-293 cells lines
expressing either human 122 or 532 GABA-A receptor. By combining a DynaflowTM
platform integrated with a patch-clamp equipment for measurements of chloride ion flux in single
HEK- 293 cells it is possible to expose a cell only expressing one human GABA-A receptor subtype
during a few milliseconds. With short (ms) exposure time problems with desensitization are not seen
and the receptor-opening pattern is similar to the one at spontaneous GABA release. With this
technique results in the HEK-293 cells expressing 122 GABA-A receptors show concentration
response curves for GABA with an EC50 value of 4 µM and Emax 2500 pA. For 532, the EC50
value is 15 µM and the maximal effect 5200 pA.

P59
Insulin modulation of GABA-A receptors-mediated inhibition in rat hippocampus
JIN Z, MENDU SK, BIRNIR B
Department of Clinical Science (CRC) Lund University, University Hospital MAS Ent 72 Bldg 91
Fl11, Malmö 20502, Sweden

Hypothalamic and hippocampal neurons in the central nervous system can sense and respond to
changes in metabolic signals such as insulin and glucose. GABA (gamma-aminobutyric acid) and its
receptors have been shown to have a role in the central control of metabolic homeostasis. The aim of
our study was to investigate the insulin effect on GABA-A receptors-mediated inhibition in rat
hippocampal CA1 pyramidal neurons. Rat hippocampal slices (P16-P22) were prepared and
incubated with low concentration of insulin for 1, 2, and 3 hours. Standard whole-cell recording and
quantitative PCR were performed. Insulin incubation regulated both phasic and tonic currents
mediated by GABA-A receptors in a temporal manner. PCR results from the CA1 region showed
altered mRNA expression of alpha4, alpha5 but not delta GABA-A receptor subunits. Our results
suggest that low concentration of insulin potentiates GABA-generated inhibition by modulating both
synaptic and extrasynaptic GABA currents.




                                                 130
                                             Abstracts

P60
AA29504 – a modulator of extrasynaptic GABA-A receptors
HOESTGAARD-JENSEN K1,2, VARDYA I 1, DALBY NO 2, JENSEN K 1,EBERT B2
1
  Inst. of Physiology and Biophysics, Univ. of Aarhus, Aarhus C, Denmark 2Department of
Electrophysiology, H. Lundbeck A/S, Valby, Denmark

GABA is the major inhibitory neurotransmitter in the central nervous system. The GABA-A receptor
is a chloride gating pentameric structure that can be allosteric modulated by several classes of
compounds. GABA-A receptor subtype selective compounds may provide the basis for developing
new drugs. The compound named AA29504 was characterized with respect to subtype selectivity in
Xenopus laevis recombinant expression system and in hippocampal slice electrophysiology.
AA29504 potentiated GABA currents at synaptic GABA-A receptors (a1b3y2s). The modulatory
effect of AA29504 was independent of the alpha subunit but sensitive to the presence of the gamma
subunit. However, the modulatory effect was insensitive to the benzodiazepine antagonist
flumazenil, making an interaction with the benzodiazepine site located at the interface between the
alpha and gamma subunit unlikely. GABA-A receptors containing beta2/3 were more strongly
positive modulated than beta1 containing receptors. The strongest positively modulatory effect was
seen at extrasynaptically located delta-subunit containing GABA-A receptors. When Gaboxadol, a
functionally selective extrasynaptic GABA-A agonist was used the response was strongly
potentiated by AA29504 at extrasynaptic GABA-A receptors. Whole-cell patch-clamp recordings
were made from dentate gyrus granule cells in mouse brain slices. Recordings of mIPSCs showed
that AA29504 (1 µM) significantly prolonged the mIPSC decay time constant. The GABA-A
receptor antagonist SR95531 revealed a small tonic GABA-A current in presence of AA29504.
AA29504 strongly potentiated the tonic current to Gaboxadol. In conclusion, AA29504 exerts a
unique subtype- dependent modulation of GABA and Gaboxadol responses particularly at
extrasynaptic GABA-A receptors.

P61
GABA input into orexin neurons is involved in proper maintenance of vigilance state
MATSUKI T, BETTLER B, YANAGISAWA M, SAKURAI T
Department of Molecular Neuroscience and Integrative Physiology, Graduate School of Medical
Science, Kanazawa University, Kanazawa 920-8640, Japan

The neuropeptides orexin (also known as hypocretin), produced in hypothalamic neurons, are critical
regulators of sleep and wakefulness. The symptoms and pathophysiology of the sleep disorder
“narcolepsy”, caused by an orexin deficiency provide insight into the physiological roles of orexin.
Electrophysiological studies have identified several neurotransmitters and neuromodulators that
activate or inhibit the activity of orexin neurons, and GABA is thought to be one of the major factors
that influence orexin neuronal activity. In the present study, to evaluate the physiological relevance
of GABA-mediated regulation of orexin neurons in vivo, we examined the involvement of GABA-B
receptors in the regulation of orexin neurons, by utilizing mice with a selective deletion of the
GABA-B1 subunit in these neurons. Patch clamp recording studies showed that the inhibitory
response of orexin neurons to a GABA-B agonist was completely eliminated in brain slices from the
conditional GABA-B1 knockout mice. Surprisingly, orexin neurons were also less responsive to
glutamate because the augmented GABA-A receptor-mediated inhibition increases the membrane
conductance. These observations indicate that absence of GABA-B receptors decreases the

                                                 131
                                               Abstracts

sensitivity of orexin neurons to both excitatory and inhibitory inputs. Sleep state analysis of the
conditional GABA-B1 knockout mice revealed that these mice displayed a robust fragmentation of
the sleep/waking states during both light and dark periods. These observations suggest that GABA-B
receptors in orexin neurons are essential for the normal regulation of orexin neuronal activity, and
regulation of sleep/wakefulness states.

P62
Neurosteroids 3,20 (R/S)-pregnandiols decrease offset rate of the GABA-site activation at the
recombinant GABAA receptor
WANG M-D, BORRA VB, STRÖMBERG J, LUNDGREN P, HAAGE D, BÄCKSTRÖM T
Umeå Neurosteroid Research Center, Department of Clinical Science, Obstetrics and Gynecology, 2
Department of Integrative Medical Biology, Umeå University, S-901 85 Umeå, Sweden

Neurosteroids directly modulate ligand gated ion channels such as GABAA receptors. Two such
molecules, 3-OH A-ring reduced pregnane steroids and pregnenolone sulfate (PS), inhibit
recombinant GABA-A receptor. Using a two-electrode voltage-clamp technique, we compared the
effect of 5-pregnan- 3,20(S)-diol (UC 1019), 5- pregnan-3,20(R)-diol (UC1020) and PS on the
activation onset and offset time of the recombinant GABAA receptor (rat 122L) in Xenopus
oocytes. Rapid solution changes allowed the kinetic analysis of GABA-evoked currents. Steroids
were co-applied with 30 µM GABA for 10 s, followed by a 80 s wash out period. PS (≥0.3 µM)
moderately increased the slow onset rate (kon-S) of GABA-response. PS had no significant effects on
the fast onset rate (kon-F). UC1019 and UC1020 decreased the kon-S of the GABA-response in a
concentration-dependent manner with no significant effects on the kon-F. Like PS, UC1019 and
UC1020 decreased the slow offset rates (koff-S). In addition, PS increased the fast offset rate (koff- F)
in a concentration-dependent manner, while UC1019 and UC1020 decreased koff- F. The EC50 of PS
to increase koff-F was calculated as 0.47±0.1 µM. The corresponding IC50 values of UC1019 and
UC1020 to decrease koff-F were 5.0±0.5 µM and 8.4±0.9 µM, respectively. These results suggest
differential actions of PS and 3,20(R/S)-pregnandiols on the offset time course of GABA-site
activation.

P63
Milk Sugar Test: social competition model for study of direct effect of GABAA receptor active
compounds
BENGTSSON SK, LÖFGREN M, JOHANSSON IM, BÄCKSTRÖM T
SK UNC, NUS 5B 5TR, 901 85 Umeå, Sweden

Negative mood, depression, irritability and aggression are known to occur in certain individuals due
to paradoxical effect of -amino butyric acid type A (GABAA) receptor active compounds, e.g.
alcohol, anabolic steroids and other neurosteroids, which constitutes a large problem in our society.
Allopregnanolone (allo), a progesterone metabolite and neurosteroid, is found to enhance alcohol
induced behaviour as well as being contributing factor to premenstrual disorder. The aim of this
study was to develop an animal model, based on a scenario comparable to the social stress of
patients negatively affected by (GABAA) receptor active compounds, and optimize the model in
terms of dose and interval between treatment and test. Male rats were housed in triads, within which
they form stable hierarchies where the subordinate rat is weaker in competition which leads to stress
and anxiety. In our model one smaller rat, assumed to be subordinate, was housed with two larger

                                                  132
                                             Abstracts

rats in order to heighten the effect of social competition. We found that in a 5 minutes‟ competition
for sweetened milk, Milk Sugar Test (MST), the subordinate rat acquired low access to drink.
However, after 0.5mg/kg and 1.0mg/kg allo treatment (i.v.), with a 5 minutes interval, the smaller rat
showed dose dependant increase in drinking time linked to irritability behaviours. MST thereby
constitutes a social competition model suitable for direct effects of GABAA receptor active
compounds and linked irritability behaviours. MST is also thought to display an animal scenario
comparable to the situation of persons who experience paradoxical effects of GABAA receptor active
compounds.

P64
Extrasynaptic-like GABA channels expressed in lymphocytes
KUMAR MENDU S, ÅKESSON L, JIN Z, CALCAGNILE O, CILIO CM, LERNMARK Å,
BIRNIR B
Lund University, Clinical Research Center, Entry-72, Plan-91-11, Malmö 20502, Sweden

BACKGROUND AND AIMS: GABA is the predominant inhibitory neurotransmitter in the
mammalian central nervous system (CNS) where it binds to GABA-A receptors (GABA channels)
and opens their chloride channel. Outside the nervous system, pancreatic alpha cells and
lymphocytes may express GABA channels. The GABA concentration in plasma in healthy
individuals is around 100 nM. We have shown that extrasynaptic GABA channels are activated by
submicromolar GABA concentrations. In this study we examined if CD4+ and CD8+ lymphocytes
isolated from mesenteric lymph nodes (MLN) from BB (BioBreeding) rats express GABA channel
subunits. MATERIAL AND METHODS: Lymph nodes were isolated from congenic BBlyp/lyp rats
that develop T1D spontaneously and from wildtype BB+/+ rats. Lymphocytes were isolated and
CD4+ and CD8+ cells identified and purified. Total RNA was extracted from the cells and brain
samples used as a positive control. RNA was reverse transcribed and used in quantitative RT- PCR
reactions using GABA-A subunit specific primers. RESULTS:. Cells isolated from MLN BBlyp/lyp
and BB+/+ rats, showed similar pattern of subunit expression with some variation between CD4+
and CD8+ cells. The most prominent expression was observed for alpha1, alpha6, beta3 and delta
subunits. Notably, the gamma2 subunit thought to be present in all synaptic receptors was absent. At
onset of diabetes, the relative expression of the alpha4 subunit was increased in CD4+ and CD8+
cells whereas the alpha6 expression was decreased. CONCLUSIONS: The results are consistent with
expression of high-affinity, extrasynaptic-like GABA channels in CD4+ and CD8+ lymphocytes.
Supported by: Novo Nordisk Foundation, Crawford Foundation, Swedish Research Council




                                                 133
                                              Abstracts


                      Poster session 10 – Renal and comparative physiology

P65
Plasmin stimulates the epithelial sodium channel via interaction with prostasin
SVENNINGSEN P, UHRENHOLT TR, JENSEN BL, SKØTT O
Physiology and Pharmacology, Institute of Medical Biology, University of Southern Denmark,
Odense C, Denmark

Several pathophysiological conditions are characterized by increased activity of the epithelial
sodium channel (ENaC); among them, the nephrotic syndrome. We recently identified plasmin as a
stimulator of ENaC activity in nephrotic urine, and this study was undertaken to investigate the
mechanism by which plasmin stimulates ENaC activity. Using small interference RNA (siRNA),
expression of the ENaC subunit was knocked down in the mouse cortical collecting cell line M-1,
which prevented stimulation of ENaC activity by plasmin. Recent evidence indicates that prostasin,
a glycosylphosphatidylinositol (GPI)-anchored serine protease, activates ENaC by proteolytic
cleavages of the subunit, leading to the release of a peptide from the extracellular domain. To
explore if plasmin-stimulation led to a similar response, a hexa-histidine tag was introduced in the
proposed peptide and the tagged ENaC subunit in M-1 cells. The hexahistidine tag was readily
visualized in untreated cells by fluorescence microscopy using the fluorophore NTA-Atto550;
however, treatment with plasmin abolished labeling. Using biotin-label transfer technique, we found
that plasmin interacts with prostasin on M-1 cells. Stimulation of M-1 cells with plasmin induced
cleavage of prostasin, which was inhibited by mutation of the activation-site within prostasin.
Moreover, we found that siRNA knock-down of prostasin prevented plasmin-stimulated ENaC
activity in M-1 cells. Release of GPI-anchored proteins from the cell surface of M-1 cells by
incubation with phosphatidylinositol-specific phospholipase C, inhibited the plasmin-induced
removal of the hexahistidine tagged ENaC subunit. Thus, our data suggest plasmin interacts with
prostasin, which leads to cleavage of the  subunit and activation of ENaC.

P66
The significance of prostasin for tight epithelial formation
STEENSGAARD M, SVENNINGSEN P, JENSEN BL
Institute of Medical Biology, Dep. of Physiology and Pharmacology, J.B. Winsloewsvej 21 3., 5000
Odense C, Denmark

Prostasin is a glycosylphosphatidylinositol anchored serine protease necessary for acquisition of
barrier function of epidermis (Leyvraz et al. 2005). Prostasin is expressed in adult kidney. In rat,
urine concentrating ability develops in postnatal weeks 2-3 governed by glucocorticoid. We
hypothesized that prostasin is sensitive to glucocorticoid and essential for tight junction formation in
renal collecting duct. The mouse cortical collecting duct cell line M-1 was used to study formation
of tight epithelium in vitro. M-1 cells grew on semipermeable membranes which allowed
measurements of transepithelial resistance (TER) and –voltage (TEV). Cells were seeded at
4x105/ml and developed a significant increase in TER (79.8 ohm ±3.4 to 1346 ohm ±213, p=0.0001,
n=6) and TEV (1.55mV±0.2 to 20.7mV±3.8, p=0.0005, n=6). In the same period prostasin mRNA
and protein level increased significantly in M-1 cells. Omission of the synthetic glucocorticoid
dexamethasone from the culture medium abolished the increase in prostasin mRNA and impaired the
development of TER (1346 ohm ±212.9 vs. 403.3 ohm ±44.7, p=0.0015, n=6) and TEV

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                                               Abstracts

(20.7mV±3.8 vs. 0.6 mV±0.2, p=0.0003, n=6). Addition of the serine protease inhibitor, aprotinin, to
the apical but not the basolateral side of M-1 monolayers inhibited the development of TER (1497
ohm ±183 vs. 997.2 ohm ±131, p=0.04, n=12) and TEV (19.1mV±1.5 vs. 10.2mV±1.7, p=0.0008,
n=12) relative to the controls. In a developmental series of rat kidneys, prostasin mRNA abundance
increased significantly between postnatal days 0- 20, when urine concentrating capacity increases ~5
times. Serine protease activity and glucocorticoids are necessary for development of tight collecting
duct epithelium in vitro. Prostasin is a candidate protease that displays sensitivity to glucocorticoids.
Leyvraz, C. et al. 2005. J. Cell Biol., 170, 487- 496.

P67
Lithium impairs kidney development and inhibits glycogen synthase kinase-3β activity
KJAERSGAARD G, MADSEN K, MARCUSSEN N, CHRISTENSEN S, JENSEN BL
Institute of Medical Biology, Department of Physiology and Pharmacology, J.B. Winsløwsvej 21 3.,
5000 Odense C, Denmark

Lithium (Li+) is used to treat affective disorders. Prolonged use in adults results in polyuria and
ultimately renal fibrosis. Early in life, kidney maldevelopment has been described (Christensen et al.
1982). In kidney, Li+ inhibits Glycogen Synthase Kinase-3β (GSK-3β) by increased phosphorylation
on serine9 (s9) (Rao et al. 2005). In the present study we hypothesized that Li+ leads to damage of
the developing kidney through inhibition of GSK-3β. We studied the expression of GSK-3β in the
developing rat and human kidney and the phosphorylation state during postnatal development and
after Li+ treatment. Li+ was given through the chow (50 mmol Li+/ kg chow) to female Wistar rats
with litters reduced to 8 pups, in the postnatal (P) days P0-P14 or P7- P30. Pups were sacrificed and
kidney tissue was collected for molecular analysis or perfusion fixed for immunohistochemistry. At
P30, Li+- treated rat pups were polyuric and the kidneys exhibit dilated pelvis with medullary
atrophy. During postnatal kidney development, GSK-3β mRNA was stably expressed in kidney
cortex and medulla, whereas GSK-3β and pGSK-3β-s9 protein abundances decreased significantly
(at P28: 10 and 15 % of levels at P0, P<0.0001, n=6). Li+- treatment increased pGSK-3β-s9
significantly whereas total GSK-3β expression was unaltered. Immunohistochemical analysis for
GSK-3β and pGSK- 3β-s9 showed labeling associated primarily with the entire collecting duct
system both in adult and fetal human kidney and postnatal rat kidney. In conclusion, Li+ leads to
kidney damage in the postnatal period and inhibits GSK-3β in renal tissue. GSK-3β activity could be
necessary for proper kidney development. Christensen, S., Ottosen, P.D. & Olsen, S. 1982. Acta
Pathol Microbiol Immunol Scand [A] 90, 257- 267. Rao, R., Zhang, M.Z., Zhao, M., Cai, H., Harris,
R.C., Breyer, M.D. & Hao, C.M. 2005. Am J Physiol Renal Physiol 288, F642-F649.

P68
Dihydropyridine and ryanodine receptors in avian skin – what for?
PELTONEN LM, MÄNTTÄRI S
Department of Biomedicine/physiology, Biomedicum Helsinki, POB 63, 00014 University of
Helsinki, Finland

Recent evidence in pigeons and chickens show that epidermal cornified cells sequester calcium
(Ca2+) and that avian skin may inclose microenvironments with variable ionic composition. We
tested our hypothesis that skin may function as a reservoir or a secretory pathway for Ca2+ by finding
out whether appropriate cellular mechanisms exist for these functions in chickens. For calcium

                                                  135
                                              Abstracts

influx, the densities of dihydropyridine receptors (DHPRs), and for intracellular Ca2+ release,
densities of ryanodine receptors (RyRs) were examined with high affinity (-)-enantiomers of DHP
and Ry labeled with fluorophores. Activity of ionic Ca2+ was measured in plasma and in
extracellular fluid (SBF), collected by suction blister technique, and utilization of Ca2+ was
investigated by measuring the activity of the enzyme alkaline phosphatase (ALP). Our results
showed that both DHPRs and RyRs are present in all skin layers, however, the receptor densities
being the highest in the surface. The densities were higher in males, particularly in the dermis and in
mid epidermis. However, a reduction in the dietary Ca2+ decreased the densities to the same level as
in females. Spatially, RyRs seem to be located in the periphery of the sebokeratinocyte, indicating
proximity with DHPRs. The ALP activities were always lower in SBF in both genders. In females,
both plasma and SBF activities increased after coming out of lay, probably indicating increased
osteoblast activity related to the reformation of structural bone. We conclude that appropriate
cellular mechanisms for Ca2+ influx and release are present in the skin of female and male chickens,
and that higher densities in the males imply increased capacity for Ca2+ influx and intracellular
processing. Based on these results in avians, our conception of the functions of the skin need to be
reevaluated.

P69
Hypoxia and adenosine in the epaulette shark
RYTKONEN KT, RENSHAW G, NIKINMAA M
Laboratory of Animal Physiology, University of Turku, 20014 Turku, Finland

The Epaulette shark is a hypoxia-tolerant representative of Elasmobranchs, a group of vertebrates
that diverged from the lineage leading to tetrapods and teleost fishes around 450 million years ago.
In hypoxia the vertebrates normally show an increase in extra- cellular adenosine concentration.
Since adenosine is an inhibitory regulator of nerve function, such an increase will play a role in the
hypoxia-induced inactivation of all tissues. The cellular effects of adenosine are mediated via
adenosine receptors. Some adenosine receptors respond to hypoxia in mammals, but their presence
and function have not been studied in detail in water-breathing animals, even though aquatic animals
experience hypoxia more often and more severely than terrestrial animals. We exposed epaulette
sharks to hypoxia either directly or after previous exposure to hypoxic conditions and collected
brain, eye, gill and heart tissues. In ongoing experiments we use qPCR to evaluate if changes in the
transcription occur in selected genes of hypoxia- inducible factor dependent pathway including
adenosine receptors.

P70
Influence of acute hypoxia on antioxidant system of non-pregnant and pregnant rats
TROFIMOVA LK, GRAF AV, DUNAEVA TY, MASLOVA MV, KRUSHINSKAYA YV,
BAYZHYMANOV AA, GONCHARENKO EN, SOKOLOVA NA
Leninskiye gory, 1/12, Biological faculty, Department of human and animal physiology, Moscow,
Russia

Pregnancy is attended by the raised risk of oxidative stress. Pregnant organism could also get
different stress situations, for example, hypoxia. We studied the influence of pregnancy (period of
early organogenesis) and acute hypobaric hypoxia (AHH) on rats‟ antioxidant system (AOS). In
experimental groups non-pregnant rats and rats on the 9-10th day of pregnancy were subjected to

                                                 136
                                             Abstracts

AHH in a pressure chamber (oxygen pressure corresponded to 11 500 meters above sea-level).
Impact lasted before stop of rat‟s breath, but no more, than 10 min. Each experimental group had
own control not subjected to AHH. Superoxide dismutase activity (SOD, one of the most important
enzymes of AOS) and concentration of substance reacted with tiobarbiturate acid (TBA-active
products, the markers of oxidative stress), determined 24 h after stress in the blood plasma by
spectrophotometric analysis were the criteria of AOS-work. Control pregnant females showed
32.9±2.6% higher SOD-activity and the same concentration of TBA-active products than control
non-pregnant females. Thereby pregnancy mobilizes AOS-system. This mobilization could be the
cause of unchanged level of oxidative stress markers. Pregnant rats after AHH had 13.0±4.0% higher
SOD-activity and 11.1±4.2% more TBA-active products, than control pregnant rats. The non-
pregnant females from experimental and control groups had no differences from each other. We
therefore can conclude that AHH has more prolonged consequences for AOS and oxidative status of
pregnant rats than non-pregnant.

P71
Regulation of blood pressure during head movement in the anaesthetized giraffe
BRØNDUM ET, WANG T, HASENKAM JM, NYGAARD H, SECHER NH, PETERSEN KK,
BUHL R, AALKJAER C
Institute of Physiology and Biophysics University of Aarhus Ole Worms Allé 1185 DK-8000 Aarhus
C, Denmark

The giraffe experiences great cardiovascular challenges. When it lowers its head to drink, arterial
pressure at the brain is expected to increase dramatically. This study investigated how blood pressure
is regulated when lowering the head of anaesthetized giraffe. METHODS: We measured pressure by
insertion of catheters at the central part of the carotid artery and jugular vein of 5 anaesthetized
spontaneously breathing giraffes, suspended in upright position. Changes in jugular cross sectional
area were visualized by ultrasound (US). RESULTS: When the giraffes head was lowered below
heart level, the central arterial pressure (CAP) decreased from 205±14 mmHg to 139±18 mmHg, and
central venous pressure (CVP) fell 2.9±1.2 mmHg. US images revealed significant increase of the
cross section of the cranial part of the jugular veins from 0.12±0.04 cm2 to 3.16±0.59 cm2, and thus
an accumulation of estimated 2.5L of blood. CONCLUSION: When the giraffe lowers its head,
central blood volume falls and the associated reduction in cardiac filling lowers CAP by 60 mmHg.
This may contribute to protection of the brain capillaries. When the giraffe lifts the head, the vein
collapses and the accumulated blood is returned to the central circulation, increasing preload. Our
study shows that the Starling mechanism may be involved in protecting the giraffe‟s brain when
lowering the head.




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                                             Abstracts


                                 Poster session 11 – Other topics

P72
Effect of prolyl-glycyl-proline (PGP) peptide on mast cells activity in vitro
BONDARENKO NS
Dept. Human & Animal Physiology, Biology Faculty, Moscow State University, Leninskie Gori 1,
build.12, Moscow, 119991, Russia

It is known, that stress and inflammation lead to disorders in rats mesenteric microcirculatory
system. These disorders are connected with mast cells (MC) activation. Activation of MC by
compound 48/80 also led to microcirculatory system disorders. MC stabilization by ketotifen
decreased them. Proline containing peptides (PGP, PG, GP) had a protective effect in
microcirculatory dysfunction under conditions of inflammation and stress. Mechanism of peptides
protective action is not clear. This action can be connected with MC activation and may be direct as
well as indirect. The effect of PGP on rats isolated peritoneal mast cells activity (secretion of
enzyme -hexosaminidase, %) in vitro was investigated. We used two different MC activators –
synacten (ACTH1-24, Novartis) and compound 48/80 (Sigma). Synacten (2∙10-5 M) increased MC
secretion of -hexosaminidase (norm. 19.31±1.32, syn. 25.04±1.04), and MC incubation with PGP
(6∙10-5 M) prevented it (21.81±0.93). Compound 48/80 also increased -hexosaminidase secretion
(50.32±3.1). But PGP didn‟t prevent MC activation by compound 48/80 (46.8±6.62). It is possible
that effect of PGP on mast cells activity depends on the way of their activation.

P73
Photoplethysmographic measurement and pulse wave analysis
HUOTARI MJ, YLIASKA N, MAATTA K, KOSTAMOVAARA J, LANTTO VE
Department of Electrical and Information Engineering, University of Oulu PO Box 4500 FIN-90014
Oulu, Finland

Photoplethysmography (PPG) has been used for long time to measure physiological phenomena in
finger, ear, toe and ear tissue volume with pulse and respiration. Unlike strain gauge
plethysmography which measures only mechanical phenomena, PPG measures both mechanical and
optical phenomena inside of the tissues producing nice waveform series in the measurements. We
have developed both electronics and sensors for accurate PPG measurements. Afterwards pulse
waveform analysis is a convenient, inexpensive, and non-invasive diagnosis method in the
connection of PPG. Decomposing of pulse measures has to acquire and record pulse waveforms by
an optoelectronic amplifiers and sensors firstly from a resting human, and then analyze these pulse
waveforms. The pulse waveforms of the PPG measurements from the human forefinger and the
second toe blood vessels have been shown to fit with many functional forms, and especially the
functional form of the log-normal density function. We show that a log-normal density function
(distribution) can be fitted with very good accuracy to the PPG waveforms using four log-normal
components. OriginTM softwave was used in this primary analysis task. As a measurement result we
received PPG pulse waveforms which are usually composed of four primaries; percussion wave,
tidal wave, dicrotic wave, and pre-ejection wave. According to these primaries we can extract the
parameters for pulse wave velocity and especially looking forward to arterial stiffness index in
young and elderly persons. As the person age increased, the width of the finger PPG pulse increased
and smoothed which is consistent with prolongation of the time required for transmission of the

                                                138
                                              Abstracts

pulse wave at the level of the finger vessels. Other finger PPG waveform parameters, like the area,
rise and fall time, are calculated with the relevant personal parameters.

P74
Smoking habits among university students in Tartu, Estonia and Oulu, Finland
KINGISEPP PH, HEIKKINEN R, KIVASTIK J, NÄYHÄ S
Department of Physiology, University of Tartu, Ravila 19, Tartu 50411, Estonia

Smoking habits among university students in Tartu, Estonia (819 males, 1470 females), were
surveyed in 1992-93 with a questionnaire, comparisons being made with a similar study performed
in Oulu, Finland (2846 males, 3084 females) during 1990. The time of the survey was marked by
Estonia´s transition to an independent economy and western-type cigarettes. In Tartu, 31.0% of
males and 14.0% of females smoked regularly (with 19.7% of males and 5.2% of females smoking
daily), whereas in Oulu those percentages were 22.6% and 15.9% (with 13.5% and 7.6% smoking
daily), respectively. The more pronounced sex differences in smoking in Tartu than in Oulu point to
more traditional smoking patterns in Estonia. The percentages of ex-smokers were lower in Tartu
(16.9% in males, 17.7% in females) than in Oulu (28.4% and 33.5%). In Tartu the percentage of
daily smokers increased by age, from 9.6% in 18-year-old males to 30.6% in males older than 23
years, the corresponding percentages in females being 2.3% and 11.8%. Such a trend was not seen in
Oulu. We also noticed that especially female students in Tartu started smoking at the older age than
in Oulu. There were also wide variations between the faculties, in Tartu there were more smokers in
the faculties of Theology, Philosophy and Medicine, the lowest percentage was in the Faculty of
Exercise and Sport Sciences. Surprisingly, 27.1% of male and 5.9% of female medical students in
Tartu reported current daily smoking. In Oulu the biggest percentage of daily smokers was in the
Humanities and the smallest in Medicine (7.0% of male and 5.1% of female medical students). A
follow-up survey is planned for 2009 to find out whether the differences between Oulu and Tartu
smoking habits have disappeared.

P75
Enhanced cold-induced vasoconstriction (Raynaud’s phenomenon) is the sign of disadaptation
to cold
GERASIMOVA L
Department of Human and Animal Physiology, Lenin Str., 33 Petrozavodsk 185910 Russian
Federetion

The objective of the study was to assess the rate of cold-associated symptoms related to cold-
induced vasoconstriction, including Raynaud‟s phenomenon, in subjects with different adaptability
to European North, and to correlate it to functional status of major physiological systems. It has been
found that the rate of symptoms based on the cold-induced vasoconstriction, is significantly
increased in migrants at the initial period of acclimatization to Nordic environment, and also in the
subjects with chronic somatic pathology. Enhanced cold- induced vasoconstriction was shown to
correlate with sympathetic skin response that indicates the lack of cholinergic, and the essence of the
adrenergic control. Also, enhanced cold-induced vasoconstriction and cold dyspnea were
accompanied by changes of pulmonary ventilation and by impairment of sensory and motor nerve
conduction. In conclusion, the phenomenon of enhanced cold-induced vasoconstriction appears to be



                                                 139
                                              Abstracts

the sign of disadaptation to cold and also the risk factor of health impairment in the Nordic
environment.

P76
Neuronal NOS contributes to long-term facilitation of neck muscle nociception in mice
RISTIC D, ELLRICH J
Medical Physiology Group, Center for Sensory-Motor Interaction, Department of Health Science
and Technology, Medical Faculty, Aalborg University, DK-9220 Aalborg, Denmark

Administration of α,β-meATP (ATP) into neck muscles facilitates brainstem nociception in
anesthetized mice. The unspecific nitric oxide synthase (NOS) blocker L-NMMA inhibits and
reverses this facilitation. The present study addressed hypothesized involvement of the neuronal
NOS (nNOS) isoenzyme in ATP-mediated neck muscle facilitation. Bilateral infusion of ATP (1
µM, 25 µl; Sigma- Aldrich, USA) into semispinal neck muscles was performed in anesthetized mice
(n=18). Impact of neck muscle nociception on sensory brainstem processing was electro-
physiologically monitored via the jaw-opening reflex (JOR). The JOR was elicited by electrical
tongue stimulation and monitored for at least 90 min after ATP infusion (control). Different dosages
of highly selective nNOS inhibitor NPLA (0.5, 1, 2 mg/kg, Tocris, USA) were intraperitoneally
injected 30 min before or 90 min after ATP infusion. After sole ATP infusion JOR increased by
203±18 % (mean±sem, p<0.001). Preceding NPLA (2 mg/kg) prevented facilitatory ATP effect for
at least 90 min. After preceding injection of 0.5 mg/kg and 1 mg/kg NPLA ATP effect decreased to
95±12 % and 56±7 %, respectively. Subsequent application of 2 mg/kg NPLA did not reverse
established ATP-evoked reflex facilitation. ATP reliably induced long-term facilitation of neck
muscle nociception. Preceding NPLA application blocked this facilitation in a dose- dependent
manner. Subsequent nNOS inhibition did not affect reflex facilitation. Thus, nNOS probably plays a
role in induction but not maintenance of nociceptive facilitation in the brainstem. Inducible or
endothelial NOS isoenzymes are putative candidates for effect maintenance. These results point to a
major role of NOS isoenzymes in neck muscle nociception.

P77
Gender differences in cooling-induced changes in muscular activity
SORMUNEN E, RISSANEN S, OKSA J, PIENIMÄKI T, REMES J, RINTAMÄKI H
Centre for Arctic Medicine, Thule Institute, University of Oulu, Finland

In thermoneutral conditions with identical work tasks, women generally have higher muscular
activity and higher prevalence of musculoskeletal disorders of the neck and upper extremity
compared to men. The objective of this study was to investigate the effect of gender on cooling-
induced changes on muscular activity during 2 hours of repetitive manual work at the exposures of
19 ºC (thermoneutral, TN) and 4 ºC (cold, C). There were eight men and eight women as test
subjects. Their physical characteristics were (men/women): age 25±4 / 23±3 years, height 179±5 /
163±4 cm, body mass 75±12 / 57±4 kg and body fat 14.2±5.0 / 24.3±3.3 %. Each subject
participated once to thermoneutral and cold conditions in a random order. Muscular activity was
studied in eight muscles in upper extremity and the shoulder region by surface electromyography.
Rectal and skin temperatures (from 15 sites) were measured continuously during the study. During
the work at C, mean skin temperature was lower both in men and women compared to work at TN
(p<0.05). Women had 0.7 ºC and 0.5 ºC lower mean skin temperature compared to men at C and TN,

                                                 140
                                             Abstracts

respectively (p<0.05). Muscular activity in upper extremity and the shoulder region was higher in
women compared to men: 6-57 % higher at C and 8-70 % higher at TN (not significant - p<0.05).
Interestingly, cooling increased muscular activity more in men (3.6-30.4 %) compared to women
(2.3-15.1 %) although the level of muscular activity in men remained still lower than in women. In
conclusion, repetitive work at C increases muscular activity, in both genders and especially in men,
compared to similar work at TN. The observed gender differences in muscular activity should be
considered when evaluating work promotion in cold conditions.

P78
Performing cognitive task increases oxygen consumption during low intensity work at
thermally neutral and cold environment
TROUBAT N, DUGUE B, HYRKAS H, OKSA J
Laboratory of Exercise-Induced Physical Adaptations, EA 3813, University of Poitiers, France

The aim of this study was to evaluate if performing cognitive task while doing low intensity
repetitive work is able to increase oxygen consumption (VO2) in two different ambient temperatures.
Ten healthy men performed continuous wrist flexion-extension repetitive work at 10% maximal
voluntary contraction (MVC) for one hour. During the last 30 minutes they also performed a
cognitive task (Verbal Span test). Participants performed these tasks in thermoneutral (25°C) and
cold (5°C) ambient temperature. Oxygen consumption (VO2) was measured continuously throughout
the experiments. We observed that skin temperature was stable in thermoneutral (33.0±0.1°C) but
significantly decreased in cold (26.1±0.2°C, p≤0.05). Oxygen consumption during repetitive work
was 4.2±0.2 and 5.1±0.1 ml.min-1.kg-1 at 25 and 5°C, respectively. Adding the cognitive task
increased VO2 to 4.6±0.2 and 6.5±0.4 ml.min-1.kg-1 (p≤0.05). The percentage increase in VO2 due to
adding cognitive task was 9 and 21% at 25 and 5° C, respectively. The combination of repetitive
work and cognitive task at 5°C was able to increase VO2 by 35% (p≤0.05) as compared to
performing only repetitive work at 25°C. In conclusion, performing cognitive task together with low
intensity work is able to significantly increase VO2, especially in cold ambient temperature. This
should be taken into account while giving instructions to workers performing similar tasks at low
ambient temperatures.




                                                 141
                                                          Author index

AALKJAER C ............................................ 137                 BORRA VB ................................................ 132
AALKJAER C .............................................. 88                BORZYKH AA .......................................... 115
ABE A .......................................................... 76         BOUSHEL R ................................................ 93
ABRAHAMSSON T .................................... 51                       BRAKEBUSCH C........................................ 37
ABRAMOCHKIN DV ........................... 87, 97                           BREIVIK L .................................................. 96
ACAR H ..................................................... 107            BROERE N ................................................ 121
AHTIALANSAARI T ................................ 108                        BROWN AR ................................................. 78
AHTIKOSKI A ............................................ 94                 BROWN RD ................................................. 66
AHTIKOSKI AM ....................................... 116                    BRUTON JD .............................................. 118
AIRAKSINEN MS ..................................... 120                     BRØNDUM E .............................................. 75
AKINFIEVA OV ....................................... 101                    BRØNDUM ET .......................................... 137
ALHONEN L ..................................... 107, 108                    BUHL R ...................................................... 137
ALITALO K ................................................. 35              BURAVKOV SV ....................................... 115
AMASHEH S ............................................... 49                BURNIER M ................................................ 60
ANDERSLAND K ..................................... 105                      BÜYÜKBAŞ S ........................................... 109
ANDERSSON DC ....................................... 87                     BÄCKSTRÖM T .......................... 79, 130, 132
ANDERSSON M ......................................... 51                    CALCAGNILE O ....................................... 133
ANDRADE D............................................... 76                 CARLSSON PO ........................................... 91
ANDREEV-ANDRIEVSKII AA ............... 115                                  CARLSTRÖM M ......................................... 66
ANTTILA K ............................................... 113               CHEN M ............................................. 121, 122
ARNER A..................................................... 36             CHRISTENSEN S ...................................... 135
ARTEMIEVA MM .................................... 101                       CHRISTOFFERSSON G ............................. 91
ATALAY M ............................................... 114                CILIO CM .................................................. 133
AURIOLA S ............................................... 107               CINAR A .................................................... 121
AYAZ M .................................................... 125             CRISTESCU A ............................................. 96
BADMAEVA KE....................................... 120                      CURRY FE ................................................... 36
BARAEVA ZV .......................................... 120                   CURRY FR ................................................ 100
BAYZHYMANOV AA ............................. 136                            DALBY NO................................................ 131
BEBENECK M ............................................ 53                  DALIGER IL .............................................. 101
BECH C ........................................................ 72          DANIEL H ................................................. 122
BELELLI D .................................................. 78             DE JONGE HR........................................... 121
BELVİRANLI M ....................................... 109                    DE JONGE HR........................................... 122
BENGTSSON MW ...................................... 83                      DE NICOLA AF........................................... 78
BENGTSSON SK ...................................... 132                     DIAZ V ....................................................... 112
BERGERSEN LH ........................................ 51                    DIBAJ P........................................................ 91
BERNTSEN HH ........................................... 72                  DIETTERLE VY ........................................ 101
BETTLER B ............................................... 131               DJATCHKOVA I ....................................... 127
BIEDERER B ............................................. 121                DONNER K .................................................. 58
BIRNIR B ..................................... 44, 130, 133                 DONOWITZ M .......................................... 121
BLOMHOFF HK ....................................... 105                     DONOWITZ M .......................................... 122
BOISSEAU N ............................................. 112                DRINGENBERG U .................................... 122
BONDARENKO NS .................................. 138                        DRUZIN M ................................................ 128
BOOTMAN MD .......................................... 69                    DUGUE B .......................................... 112, 141
BOROVIK A ................................................ 93               DUNAEVA TY .......................................... 136
BORRA R............................................... 46, 93               EBERT B .................................................... 131

                                                                      142
ELINDER F ................................................. 41              HARZHEIM D ............................................. 69
ELLRICH J .......................................... 90, 140                HASENKAM JM ....................................... 137
ELMOSE SF ................................................ 90               HASSINEN M ........................................... 103
ELTZSCHIG HK ......................................... 55                   HAUG TM ................................................. 105
ENEA C ..................................................... 112            HAVERINEN J ............................................ 69
ENGELHARDT R ....................... 84, 121, 122                           HEIKKINEN AE ......................................... 45
ENGELKE K ............................................... 53                HEIKKINEN R .......................................... 139
FANDREY J ................................................ 65               HEIKKINEN R .......................................... 117
FAUCONNIER J ......................................... 87                   HEINONEN A ........................................... 115
FEARNLEY C ............................................. 69                 HEINONEN I ............................................... 93
FLEMSTRÖM G ......................................... 83                    HEINONEN M .......................................... 107
FOLKOW LP ............................................... 72                HELGELAND E .......................................... 96
FOX MA .................................................... 116             HELGELAND G.......................................... 96
FREDHOLM BB ......................................... 56                    HELLMAN B .............................................. 56
FROMM M .................................................. 49               HELLSTEN Y.............................................. 93
FÖLSCH UR .............................................. 107                HENRIKSNÄS J .......................................... 91
GACO V ...................................................... 48            HERD MB.................................................... 78
GAINULLINA DK .............................. 88, 101                        HERZIG K-H ....................... 83, 107, 108, 120
GERASIMOVA L ..................................... 139                      HESSELINK M ........................................... 94
GERGERLİOĞLU HS............................... 109                          HICKS JW ................................................... 76
GINIATULLIN R ...................................... 126                    HIGAZI D .................................................... 69
GIWERCMAN A......................................... 82                     HILLESHEIM J ......................................... 121
GOLICHENKOV VA ................................ 104                         HOESTGAARD-JENSEN K ..................... 131
GOMAZKOV OA ....................................... 98                      HOGEMA BM ........................................... 121
GONCHARENKO EN .............................. 136                           HOGEMA BM ........................................... 122
GOUISSEM S ............................................ 130                 HOLM L .............................................. 84, 119
GOVARDOVSKII VI .................................. 57                       HOLMER I ................................................ 112
GRAF AV .................................................. 136              HOX H ....................................................... 110
GRAPENGIESSER E .................................. 56                       HRISTOVSKA A-M.................................... 67
GREBENYUK S .......................................... 42                   HUBBARD A ............................................ 121
GREGOR M................................................. 48                HUHTANIEMI I .......................................... 81
GROOS S ..................................................... 84            HULTSTRÖM M ......................................... 85
GRÄNDE P-O ............................................. 75                 HUOTARI A .............................. 107, 108, 120
GUENNOUN R ........................................... 78                   HUOTARI MJ............................................ 138
GUNDERSEN V ......................................... 51                    HYRKAS H ............................................... 141
GUNN BG ................................................... 78              HYYPPÄ S ................................................ 114
GÜL İ ......................................................... 109         HÄKKINEN A ........................................... 115
GÖKBEL H ............................... 107, 109, 125                      HÄRKÖNEN P .......................................... 109
HAAGE D ............................................ 79, 132                IIJIMA T .................................................... 105
HAAPALAHTI J ....................................... 117                    ISAKSSON M............................................ 130
HAAVISTO M-L ......................................... 63                   ISSAKAINEN J ......................................... 115
HAKALAHTI A ........................................ 103                    IVERSEN BM.............................................. 66
HAKKO H ................................................... 69              IVERSEN BM.............................................. 85
HANCU M ................................................... 96              JAKUS J ..................................................... 114
HANSE E ..................................................... 51            JANSSON EÅ ............................................ 119
HANSEN PB ............................................. 100                 JEDSTEDT G .............................................. 83
HANSEN PB ............................................... 67                JENSEN BL ............................................... 100
HARTIALA J............................................... 46                JENSEN BL ........................... 67, 85, 134, 135

                                                                      143
JENSEN K ............................................ 44, 131              KOSKINEN S .............................................. 94
JIN Z ............................................. 44, 130, 133           KOSTAMOVAARA J ................................ 138
JOHANNESEN MD..................................... 90                      KOZHEVNIKOVA VV ............................... 97
JOHANSEN GF ......................................... 105                  KRABBENHOFT A ..................................... 84
JOHANSSON IM ....................................... 132                   KRABBENHÖFT A ................................... 121
JOHANSSON I-M ..................................... 130                    KRABBENHÖFT A ................................... 122
JOHANSSON S ................................... 42, 128                    KRUSHINSKAYA YV .............................. 136
JOHANSSON SM ........................................ 56                   KUDRYASHOVA TV ................................. 88
JOKELAINEN J ......................................... 109                 KUDRYAVTSEVA OS ............................... 98
JONASSEN A .............................................. 96               KUMAR MENDU S .................................. 133
JUNG K ........................................................ 90         KUNWALD MR .......................................... 90
JUUTI A-K ................................................. 109            KUPARI J ................................................... 120
JÄMSÄ T ................................................... 117            KUZMIN VS .......................................... 87, 97
JÄNNE J ..................................................... 107          KYRYLENKO O ....................................... 108
JÄNNE OA................................................... 81             KÄKELÄ R .................................................. 73
JÄNTTI V ................................................... 126           LAAKSO M ............................................... 109
KALENCHUK VU ............................ 101, 115                         LABOMBARDA F ...................................... 78
KALENDER WA ......................................... 53                   LAI E ............................................................ 66
KALLIO M ................................................. 116             LAMBERT JJ ............................................... 78
KALLIOKOSKI K ....................................... 93                   LAMMENTAUSTA E ............................... 115
KAMENSKY AA....................................... 128                     LAMPRECHT G .......................................... 48
KARVONEN E .......................................... 126                  LANTTO VE .............................................. 138
KASKINORO K ........................................... 93                 LAPSHINA KV .......................................... 129
KATZ A ........................................... 56, 87, 94              LATVANLEHTO A ................................... 116
KAUPPINEN M ......................................... 126                  LEHENKARI P .......................................... 106
KAUPPINEN P .......................................... 126                 LEHTONEN S ............................................ 106
KEINÄNEN-KIUKAANNIEMI S ............. 109                                  LEPPÄ E ...................................................... 79
KELBER A ................................................... 57            LEPPÄLUOTO J ........................................ 117
KEMMLER W ............................................. 53                 LERNMARK Å .......................................... 133
KEMPPAINEN J .......................................... 93                 LESKELÄ TT ............................................ 127
KHIROUG L .............................................. 126               LIERE P ........................................................ 78
KIEHNE K ................................................. 107             LILJA M ..................................................... 103
KIM PA ...................................................... 128          LIND O ......................................................... 57
KINGISEPP PH ......................................... 139                 LINDEN AM ................................................ 79
KINNUNEN S ............................................ 114                LINDROOS M ............................................. 93
KIVASTIK J ............................................... 139             LUNDBERG J ...................................... 84, 119
KIVIRANTA I ........................................... 115                LUNDGREN P ............................. 79, 130, 132
KJAERSGAARD G ................................... 135                      LUOMALA M ............................................ 112
KLAUSZ K ................................................ 107              LÜNNEMANN M ...................................... 121
KOCHER O ................................................ 121              LÖFGREN M ............................................. 132
KOCHER O ................................................ 122              LØKKA G .................................................. 100
KOLMANNSKOG O ................................. 110                        MAATTA K ............................................... 138
KOMULAINEN J ........................................ 94                   MADSEN K ............................................... 100
KOPYLOVA GN ....................................... 120                    MADSEN K ......................................... 85, 135
KORPELAINEN R .................................... 117                     MAKOWSKA A .......................................... 90
KORPI ER .............................................. 45, 79             MALININA E ............................................ 128
KOSKELA E ................................................ 73              MANNS MP ............................................... 122
KOSKI A .................................................... 116           MANNS MP ................................................. 84

                                                                     144
MAPPES T................................................... 73             NÄßL A...................................................... 122
MARCHENKO D ........................................ 93                    NÄYHÄ S .................................................. 139
MARCUSSEN N ................................. 85, 135                      OHARA S .................................................. 105
MASLOVA MV ........................................ 136                    OIKARAINEN T ....................................... 116
MATSUKI T .............................................. 131               OIKARI S .................................................. 107
MAUGHAN RJ ........................................... 39                  OJA SS ....................................................... 127
MAURIALA T........................................... 107                  OJALA R ................................................... 115
MEDVEDEV OS ....................................... 101                    OKSA J ........................ 40, 112, 113, 140, 141
MEDVEDEVA NA ....................... 97, 98, 101                           OKSALA N................................................ 114
MEIGAL A ................................................ 125              OKUDAN N .............................. 107, 109, 125
MELANDER O ........................................... 60                  OLLILA K ................................................. 126
MENDU SK ............................................... 130               OLSEN SJ .................................................. 123
MIKKOLA I .............................................. 109               OPPEGÅRD C ........................................... 105
MILLET C ................................................. 112             OTTAVY M ............................................... 112
MILLS S ...................................................... 73          PAAJANEN V ................................... 103, 104
MITCHELL EA ........................................... 78                 PARSONS K ................................................ 39
MOCHALOV SV ................................ 88, 102                       PATZAK A .................................................. 66
MOE B ......................................................... 72         PAUNIO T ................................................... 63
MOEN I ..................................................... 100           PEDERSEN M ............................................. 85
MOKKONEN M .......................................... 73                   PEITSO A .......................................... 109, 113
MOZOS I ..................................................... 96           PEKURINEN T.......................................... 104
MRDALJ J ................................................... 96            PELTO-HUIKKO M.................................. 127
MULLIEZ J ............................................... 112              PELTONEN J .............................................. 93
MULTANEN J........................................... 115                  PELTONEN LM ........................................ 135
MUSTONEN A-M....................................... 73                     PERSSON AEG ........................................... 66
MÄKELÄ K................................................. 83               PETERSEN KK ......................................... 137
MÄKELÄ KA ............................................ 108                 PETERSSON J ............................... 67, 84, 119
MÄKINEN T ............................................. 113                PETERSSON J ............................................. 48
MÄKINEN TM .......................................... 109                  PETÄJÄ-REPO U ...................................... 103
MÄNTTÄRI S ................................... 113, 135                    PETÄJÄ-REPO UE ................................... 127
MÄNTYSAARI M ...................................... 40                     PHILLIPSON M .................................. 84, 119
NADRIGNY F ............................................. 91                PHILLIPSON M .......................................... 91
NARUMIYA S ............................................ 67                 PIENIMÄKI T ........................................... 140
NEUSCH C ............................................ 84, 91               PIETILÄ M ................................................ 106
NIELSEN SS ............................................... 67              PIHLAJANIEMI T ...................................... 37
NIELSEN TB ............................................... 90              PIHLAJANIEMI T .................................... 116
NIEMINEN M ........................................... 110                 POGORELOVA MA ................................. 104
NIEMINEN P............................................... 73               POGORELOVA VN .................................. 104
NIKINMAA M .......................................... 136                  POPOV D..................................................... 93
NILSSON H ................................................. 88             PORKKA-HEISKANEN T.................... 55, 62
NISSEN-MEYER J .................................... 105                    POSTNIKOV AB ...................................... 101
NONTAMART N ........................................ 99                    POZDNEV VF ............................................. 98
NORDSTRÖM K....................................... 106                     PRYAZHNIKOV E ................................... 126
NOREEN E .................................................. 72             PUCHKOVA AN ....................................... 119
NUMMELA S .............................................. 58                PUENGPAI S ............................................... 99
NUUTILA P................................................. 46              PURHONEN AK ....................................... 108
NYGAARD H ............................................ 137                 PUUMALA P ............................................. 126
NÜSING RM ............................................... 67               PUZDROVA VA ......................................... 88

                                                                     145
RANKINEN T .............................................. 53                SEIDLER U .................... 49, 84, 121, 122, 123
RASMUSSEN LE ........................................ 67                    SEMYANOV A............................................ 42
RAUSCH B ................................................ 121               SHEVELEV SA ......................................... 101
RAUSCH B ................................................ 122               SHIRREFFS SM .......................................... 39
RAUVALA E ............................................. 115                 SHULL G ................................................... 122
REED RK ................................................... 100             SIMONOVA AI ........................................... 98
REITZ M ...................................................... 90           SIMONSEN LL ............................................ 90
REMES J .................................................... 140            SINGH A .................................................... 122
RENSHAW G ............................................ 136                  SINGH AK ................................................. 121
RIEBE I ........................................................ 51         SINGH AK ......................................... 122, 123
RIEDERER B ............................................. 122                SJOBLOM M ............................................. 123
RIEDERER B ....................................... 84, 122                  SKØTT O ............................... 67, 85, 100, 134
RINTAMÄKI H ................................. 113, 140                      SMOLANDER J ......................................... 112
RISSANEN S ..................................... 113, 140                   SMOLKA AJ ................................................ 84
RISTIC D ............................................. 90, 140              SOKOLOVA NA ....................................... 136
RODERICK HL ........................................... 69                  SOLEIMANI M .......................................... 122
ROOS S ...................................................... 119           SONG PH ..................................................... 84
ROSENSHTRAUKH LV ............................. 87                           SONKAJÄRVI E ........................................ 126
ROSSI J ...................................................... 120          SOPPELA P ................................................ 110
ROTH LSV ................................................... 57             SORMUNEN E .......................................... 140
ROTTINGHAUS I ..................................... 121                     SORMUNEN R .......................................... 116
ROTTMANN S ............................................ 90                  SRISAWAT R .............................................. 99
ROUVINEN-WATT K ................................ 73                         STAEHR M ................................................ 100
RUBIO-ALIAGA I..................................... 122                     STEENSGAARD M ................................... 134
RYGH CB .................................................. 100              STEFFENS H ............................................... 91
RYTKONEN KT ........................................ 136                    STEHR AM .................................................. 90
RYTKY S ................................................... 126             STRIDH MH ................................................ 96
RØNNING B ................................................ 72               STRÖMBERG J ........................... 79, 130, 132
SAARMA M ................................................ 35                STUART G ................................................... 35
SAASTAMOINEN E ................................. 109                        STÜTZER I ................................................ 108
SAKURAI T ....................................... 108, 131                  SUKHOVA GS ............................................ 87
SALEH A ..................................................... 56            SVENNINGSEN P ..................................... 134
SALVESEN G ............................................ 100                 SÄLLSTRÖM J ............................................ 66
SALVESEN H ............................................ 110                 TAKALA T .................................................. 94
SAMONINA GE ........................................ 120                    TAKALA TES ............................................ 116
SAND O ..................................................... 105            TARANUKHIN A ...................................... 127
SAND SL ................................................... 105             TARANUKHINA E ................................... 127
SANDBERG M ............................................ 96                  TARASOVA NV........................................ 102
SANES JR .................................................. 116             TARASOVA OS .......................... 88, 101, 102
SARANSAARI P ....................................... 127                    TAUBE M ............................................ 79, 130
SCHOMBURG ED ...................................... 91                      TAXT T ...................................................... 100
SCHREIBER O .......................................... 119                  TENSTAD O .............................................. 110
SCHUBERT R ............................................. 88                 THORSEN F ................................................ 38
SCHULZKE JD ............................................ 49                 TIMLIN S ................................................... 110
SCHUMACHER M ...................................... 78                      TIMONEN M ............................................. 109
SCHÄFER J ................................................. 48              TITOVA E .................................................. 125
SECHER NH ................................................ 76               TOPPARI J ................................................... 81
SECHER NH .............................................. 137                TROFIMOVA LK ...................................... 136

                                                                      146
TROUBAT N ............................................. 141                 WESTERBLAD H ................... 70, 87, 94, 118
TRUFANOVA AV .................................... 120                       WIIG H ...................................................... 110
TU H .......................................................... 116         VINOGRADOVA OL ............................... 115
TUNCER S ................................................ 125               WISDEN W ................................................. 79
TURNER JR ................................................ 48               VISSER H .................................................. 110
TUUSA JT ................................................. 127              WOIE K ..................................................... 110
UHRENHOLT TR ..................................... 134                      VOIKAR V ................................................ 120
UMAROVA BA ........................................ 120                     VÕIKAR V ................................................ 108
UZUNOĞLU S .......................................... 107                   VON STENGEL S ....................................... 53
VAINIONPÄÄ A....................................... 117                     VORNANEN M ................................... 69, 103
WALKER MC ............................................. 41                  VOROTNIKOV AV ............................ 88, 101
WALKOWIAK J ....................................... 108                     VOSKRESENSKAYA OG........................ 128
WANG J..................................................... 122             WULFF P ..................................................... 79
WANG M..................................................... 79              WÅHLIN N.................................................. 66
WANG M-D .............................................. 132                 VÄÄNÄNEN HK ...................................... 117
WANG T.............................................. 76, 137                YAMADA T ........................................ 94, 118
VANHOUTTE P .......................................... 67                   YANAGISAWA M.................................... 131
VARDYA I ................................................ 131               YERUVA S ................................................ 122
WATSON P ................................................. 39               YLIASKA N .............................................. 138
WEINMAN EJ ........................................... 121                  ZHANG SJ ................................................. 118
WEINMAN EJ ........................................... 122                  ZHANG S-J.................................................. 94
VEKOVISCHEVA OY ............................... 79                          ZHENH W ................................................. 123
VENABLES MC ......................................... 54                    ÅKERSTEDT T ........................................... 62
VERHULST S ............................................. 72                 ÅKESSON L .............................................. 133
VERKHRATSKY A .................................... 42                       ØRTENBLAD N.......................................... 70




                                                                      147
Campus map




    148
                  Sponsors




Scandinavian Physiological Society(main sponsor)




               University of Oulu




          Finnish Physiological Society




         Ministry of Education, Finland




  Federation of The Finnish Learned Societies




                      149
Exhibitors




   150

								
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