Knjiga Sažetaka - Kongres Neuroznanosti Zadar

3. Croatian Congress of Neuroscience, September 24‐26, 2009, Zadar, Croatia

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THE THIRD CROATIAN CONGRESS OF NEUROSCIENCE September 24‐26, 2009, Zadar

Under the Patronage of the: School of Medicine University of Zagreb Croatian Institute for Brain Research

Under the Auspices of:

Ministry of Science, Education and Sport

SPONSORS

Zagrebački holding

Hrvatska lutrija

DinSar d.o.o.

Ewopharma

Oktal Pharma

Graditelj svratišta d.o.o.

International Brain Research Organisation, IBRO


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INVITED SPEAKERS

o Maja Bućan Genetics and Gene Regulation program of the Cell and Molecular Biology graduate

group, School of Medicine, University of Pennsylvania, Philadelphia, USA o Mirko Dikšić

Cyclotron‐Radiochemistry Unit, Department of Neurology and Neurosurgery, McGill University, Montreal, Canada

o Leszek Kaczmarek Laboratory for Molecular Neurobiology, Nencki Institute of Experimental Biology,

Warsaw, Poland o Nenad Šestan Department of Neurobiology, Kavli Institute for Neuroscience, Yale University School

of Medicine, New Haven, USA ADDRESS OF ORGANIZER Croatian Society for Neuroscience Croatian Institute for Brain Research Šalata 12, HR 10000 Zagreb, Croatia Phone: +385 1 4596 801, 903 Fax.: +385 1 4596 942 www.hiim.hr CONGRESS OFFICES Croatian Institute for Brain Research, School of Medicine, University of Zagreb ‐ service information ‐ Phone No: 01/4596‐801 Martina Čuljak; [email protected] 01/ 4596‐902 Željka Pavlović; [email protected] Fax. No: 01/4596‐942 Croatian Society for Neuroscience ‐ payment information ‐ Phone No: 01/4596‐868 Nevenka Filipović; [email protected]

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GENERAL INFORMATION Hotel Funimation Borik, Zadar Majstora Radovana 7 23 000 Zadar, CROATIA www.falkensteiner.com/borik Phone No: + 385 (0)23 206‐100, Fax. No: + 385 (0)23 332 065 e‐mail: [email protected] REGISTRATION AND INFORMATION DESK Thursday, September 24th, 2009 Registration and information from 17.00 ‐20.00 h (hotel lobby) Friday, September 25th, 2009 Registration and information from 8.00 ‐15.00 h (in front of the lecture hall “Adriana”) Saturday, September 26th, 2009 Registration and information from 8.00 ‐11.00 h (in front of the lecture hall “Adriana”)

http://www.hiim.hr/

mailto:[email protected]



http://www.falkensteiner.com/borik



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REGISTRATION FEE

INCLUDED IN THE REGISTRATION FEE Name badge; Congress material; Active or passive scientific participation and continuous education credits; Welcome cocktail; Farewell dinner; Coffee breaks. POSTER SESSIONS Posters will be displayed on Saturday, 26th September, from 9.00 ‐ 14.00, and divided into two poster walks. We kindly ask poster presenters to mount their posters according to the following schedule: posters for Poster walk I are to be mounted during Friday, 25th September, and replaced during coffee break Saturday 26th September by posters scheduled for Poster walk II. Poster authors/presenters should be present at their poster board at the specified time. POSTER PRIZE Posters and student oral presentations are important scientific contributions, therefore a poster prize is established. It is awarded to three best posters out of all nine poster sessions, and the best student oral presentation. The selection will be done on the basis of scientific merit and clarity of presentation as judged by high‐ranking board made up from three members of the Scientific Committee. The awards will be announced during the lunch at Konoba „Lolina Vatrenica“ in Benkovac, 26th September. SOCIAL EVENTS All participants of the Meeting are cordially invited to join:

o Welcome cocktail with Mayor of Zadar, which will be held on Thursday, 24th September at 20.00 h at Museum of Ancient Glass in Zadar

o Farewell Dinner, which will be held on Friday, 25th September at 21.00 h in the Restaurant of hotel Falkensteiner and to visit konoba „Lolina Vatrenica“ at Benkovac for lunch on Saturday, 26th September after the closure of the Meeting.

DOCUMENTS AND BADGES Meeting documents should be collected on‐site at the registration desk. The participants are kindly asked to wear the name badges during the Meeting and in the exhibition area. FOOD AND BEVERAGE Coffee and tea during official breaks, cocktail and farewell dinner, are included in the registration fee. Lunch expenses are not covered by registration fee. LANGUAGE The official language of the meeting will be English and Croatian. CERTIFICATE OF ATTENDANCE Certificates of attendance will be distributed after the meeting. Continuous education credits will be given only to registered participants who made a requirement of CEC at registration desk. CELLULAR PHONES AND PAGERS As a courtesy to all meeting attendees and speakers, cellular phones, pagers and other electronic devices must be operated in silent/vibrate mode during session. Devices that beep, ring, etc., are strictly prohibited. Please, do not use cellular phone for conversations in the lecture hall.

EARLY (July 20th, 2009) LATE (July 20th, 2009) Society members 550 kn 650 kn Non‐members 750 kn 850 kn

Students 250 kn 350 kn Accompanying person 300 kn 400 kn


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ANNOUNCEMENT OF THE NEW JOURNAL TRANSLATIONAL NEUROSCIENCE

Dear friends and colleagues,

The company Versita will launch the Translational Neuroscience journal that will be based at Croatian Institute for Brain Research in Zagreb. The first issue of the journal is expected to come out on 1st July 2010. The aim of this international, peer‐reviewed journal is to provide a closer interaction between basic and clinical neuroscientists by publishing original papers that expand our understanding of brain structure, function, and disease, and translate this knowledge into clinical applications and novel therapies of nervous system disorders. Review articles are also encouraged and sometimes invited, as well as constructive and interesting correspondence.

Target audience of the Translational Neuroscience are both basic and clinical neuroscientists, neurologists, neuropathologists, neuroanatomists, neurosurgeons, psychiatrists, psychologists, biologists, pharmacologists, as well as students of graduate and postgraduate studies of medical and biological affiliations.

We are aware that this will be an ambitious task and that it will not be easy to start a new journal when many publishers form a strong competition. Many authors may be hesitant to publish their work in a new journal in its first years. Therefore, in the first issues, articles will be solicited mainly from the associated editors and members of the editorial board, as well as from their collaborators and colleagues. Upon achieving sufficient level of the international recognition, the content will be recruited on a peer‐review basis and editorial decisions on quality and relevance of all the submitted reports.

Versita will enter the agreement to further intensively develop the journal to a meaningful and strong publication in its field. Such development will require serious investment from Versita, as they will need to absorb the costs of electronic publishing technology (MetaPress), of an online submission system, of e‐marketing to potential authors and readers, of technical editing etc. For the initial period the contents of the journal will remain open, and from that time on they will seek to offer its distribution to Springer. Here is the current list of the Editorial Board members: FOUNDER AND EDITOR‐IN‐CHIEF: Goran Šimić, Zagreb Dr. Šimić is Associate Professor of Neuroscience, Anatomy and Clinical Anatomy at the Department of Neuroscience of the Croatian Institute for Brain Research and the Department of Anatomy and Clinical Anatomy of the Zagreb University Medical School. In the Croatian Institute for Brain Research, Dr. Šimić leads the Laboratory for Developmental Neuropathology. His laboratory has extensive expertise in brain banking (Zagreb Brain Collection) and developmental neuropathology, and has established a network for translational research in Croatia and neighboring countries aimed at early diagnosis of neurodegenerative diseases. Work Address: Goran Šimić MD, PhD, Associate Professor of Neuroscience, Anatomy and Clinical Anatomy Head of the Laboratory for Developmental Neuropathology Department of Neuroscience, Croatian Institute for Brain Research Department of Anatomy and Clinical Anatomy Zagreb University Medical School Salata 12, 10000 Zagreb, Croatia Phone lab: +385‐1‐4596820; Phone room: +385‐1‐4596807 Phone administrator: +385‐1‐4596801; Fax: +385‐1‐4596942 email: [email protected] or [email protected] URL: http://www.hiim.hr



http://www.hiim.hr/


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SENIOR ADVISORY EDITOR: Patrick R. Hof, New York Dr. Hof is the Regenstreif Professor of Neuroscience and Vice‐Chair for Translational Neuroscience of the Department of Neuroscience at Mount Sinai School of Medicine, New York. He also leads the Kastor Neurobiology of Aging Laboratories. His laboratory has extensive expertise in the pathology of neuropsychiatric disorders and has established an international reputation in quantitative approaches to neuroanatomy and studies of brain evolution. Work address: Department of Neuroscience, Mount Sinai School of Medicine, Box 1065 One Gustave L. Levy Place New York, NY 10029, USA Phone: (1 212) 659‐5904 Fax: (1 212) 849‐2510 E‐mail: [email protected] ASSOCIATE EDITORS Steven Arnold, Philadelphia Stephen Ginsberg, New York Fran Borovečki, Zagreb Elizabeth Head, Lexington Mara Bresjanac, Ljubljana Hans Kretzschmar, Munich Joseph Buxbaum, New York Nenad Šestan, New Heaven Manuel Casanova, Louisville Rohan de Silva, London Marco Catani, London Harry B.M. Uylings, Amsterdam Isidro Ferrer, Barcelona James Vickers, Hobart Panteleimon Giannakopoulos, Geneva Thomas Wisniewski, New York MEMBERS OF THE EDITORIAL BOARD Sabine Bahn, Cambridge Ivica Kostović, Zagreb Jim Barkovich, San Francisco Cynthia A. Lemere, Boston Giorgio Battaglia, Milano Paul J. Lucassen, Amsterdam Gavin J. Clowry, Newcastle Jürgen K. Mai, Düsseldorf Jeffrey Cummings, Los Angeles Glenn E. Morris, Oswestry Adrian Danek, Munich Elliot J. Mufson, Chicago Rudi A.J.O. Dierckx, Groningen Werner Paulus, Münster Richard Frackowiak, Lausanne Giulio Pasinetti, New York Yukio Fukuyama, Tokyo George Paxinos, Sydney Buzsáki György, Newark Daniela Prayer, Vienna Leszek Kaczmarek, Warsaw Eva Sykova, Prague Marijan Klarica, Zagreb Rik Vandenberghe, Leuven

WE INVITE YOU TO HELP US IN OUR EFFORTS TO MAKE TTRRAANNSSLLAATTIIOONNAALL NNEEUURROOSSCCIIEENNCCEE

AN ESTEEMED INTERNATIONAL JOURNAL BY SENDING US YOUR BEST WORK.

Thank you all for participating in this exciting endeavor,

Goran Šimić



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PROGRAM

Thursday, September 24th, 2009

17.00 ‐ 19.00 Registration at the lobby of Hotel Falkensteiner 20.00 Welcome cocktail

Friday, September 25th, 2009

Registration 8.00 ‐15.00

8.00 ‐ 9.00 breakfast 9.00 ‐ 9.10 welcome speech

Presidential symposium ‐ New Advances in Neuroscience 9.15 ‐ 10.00 Nenad Šestan: Molecular control of neocortical projection neuron identity and

connectivity 10.00 ‐ 10.45 Maja Bućan: Genomic landscape of autism spectrum disorder

10.45 ‐ 11.15 coffee break

11.15 ‐ 11.45 Mirko Dikšić: The use of tryptophan analog in studying the brain serotonergic system 11.45 ‐ 12.30 Leszek Kaczmarek: Extracellular proteolysis controlling synaptic plasticity

12.30 ‐ 15.00 lunch break

Mini symposium ‐ Approaches in Translational Neuroscience 15.00 ‐ 15.30 Branimir Jernej / Lipa Čičin‐Šain: Wistar‐Zagreb 5HT rats: an experimental model in

serotonin research 15.30 ‐ 16.30 Goran Šimić / Melita Šalković‐Petrišić: Recent findings in etiopathogenesis of

Alzheimer's disease Goran Šimić: Perspectives of translational neuroscience Melita Šalković‐Petrišić: Novel findings in streptozotocin model of sporadic Alzheimer’s disease: the origin of sporadic Alzheimer’s disease – is it brain insulin resistant state? Goran Šimić: Does Alzheimer’s disease begin in the dorsal raphe nucleus?

16.30 ‐ 17.00 Marijan Klarica: Pathophysiology of intracranial hypertension


17.30 ‐ 18.30 Annual Meeting of CSFN

21.00 Farewell Dinner ‐ Restaurant Falkensteiner

Saturday, September 26, 2009

Registration 8.00 ‐11.00

8.00 ‐ 9.00 breakfast 9.00 ‐ 11.00 Poster walk I


11.30 ‐ 12.30 Student oral presentations 12.30 ‐ 14.00 Poster walk II 14.00 ‐ 14.10 Farewell speech 16.00 LUNCH at konoba „Lolina vatrenica“, Benkovac


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POSTER SESSIONS

P1 Cerebrospinal pathophysiology and application of ultrasound P2 Clinical Neuroscience P3 Neuropharmacology P4 Molecular neuroscience P5 Neurodegenerative Neuroscience P6 Cognitive Neuroscience P7 Basic Neuroscience P8 Neurodevelopmental basis of cognitive, mental and neurological disorders P9 Special session – Case reports

POSTER WALKS

Detailed list of poster presentations with the poster numbers and student oral presentations

POSTER WALK I

Saturday, September 26th 2009 9.00 ‐ 11.00

P1 CEREBROSPINAL PATHOPHYSIOLOGY AND APPLICATION OF ULTRASOUND

PPO1 THE USAGE OF ULTRASONIC PROBE "NECUP‐2" IN NEUROENDOSCOPIC PROCEDURES Jednačak H, Paladino J, Miklić P, Vukić M, Štimac T, Klarica M

PPO2 PHYSICAL CHARACTERISTICS IN THE NEW MODEL OF THE CEREBROSPINAL FLUID SYSTEM Jurjević I, Radoš M, Orešković J, Prijić R, Klarica M PPO3 POTENTIAL ERROR IN VENTRICULO‐CISTERNAL PERFUSION METHOD FOR DETERMINATION OF CEREBROSPINAL FLUID RATE IN CATS Orešković D, Maraković J, Radoš M, Jurjević I, Klarica M

PP04 DEVELOPMENT OF NEW ULTRASONIC PROBES FOR NEUROSURGICAL AND MICROSURGICAL PROCEDURES Paladino J, Štimac T, Svilar D, Klarica M PP05 METHODS FOR MEASURING ACOUSTIC POWER OF ULTRASONIC NEUROSURGICAL DEVICE Petošić A, Ivančević B, Svilar D, Štimac T, Klarica M

P2 CLINICAL NEUROSCIENCE

PP06 PREMOTOR AND MOTOR SULCAL COMPLEX AS A POSSIBLE CANDIDATE FOR PREOPERATIVE PLANNING IN PEDIATRIC EPILEPSY SURGERY; EVIDENCE FROM PAST, PRESENCE AND FUTURE IMPLICATIONS Abdel Hadi F, Vasung L, Paladino J, Miklić P, Radoš M, Kostović I

PP07 GENE POLYMORPHISMS OF ARYLSULFATASE A IN RELAPSE REMITTING MULTIPLE SCLEROSIS: GENOTYPE‐PHENOTYPE CORRELATION AND ESTIMATION OF DISEASE PROGRESSION USING MULTIPLE SCLEROSIS SEVERITY SCORE Bačić Baronica K, Mlinac K, Vladić A, Kalanj Bognar S PP08 QUANTITATIVE EEG IN SCHIZOPHRENIA AND DEPRESSION Begić Dražen, Jokić‐Begić Nataša


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PP09 INCREASE OF BLOOD FLOW VELOCITY IN MEDIAL CEREBRAL ARTERY DURING OBSERVATION OF SUBJECT' S ARM IN A MIRROR Bene R, Lovrenčić‐Huzjan A, Ažman D, Strineka M, Budišić M, Vuković V, Rastovčan B, Demarin V PP10 THE MOST COMMON SYMPTOMS OF PINEAL GLAND CYST Bošnjak J, Miškov S, Hećimović H, Čović Negovetić R, Demarin V PP11 AGE‐DEVELOPMENTAL STAGE AND SEVERITY OF TRAUMA RELATED SYMPTOMS, ANXIETY AND DEPRESSIVE SYMPTOMS IN PARTICIPANTS WHO LOST THEIR FATHERS DURING THE WAR IN CROATIA Dijanić Plašć I, Poljarević S, Lončar M, Henigsberg N PP12 AUTISM IN CHILDHOOD‐EARLY SCHIZOPHRENIA OR SOMETHING ELSE? ATYPICAL ANTIPSYCHOTIC AS A DRUG CHOICE Dodig‐Čurković K, Čurković M, Degmečić D, Dodig‐Radić J, Radić M

PP13 SEASONALITY OF SCHIZOPHRENIA BIRTH BY SCHIZOPHRENIA TYPES Henigsberg N, Kalember P, Folnegović Šmalc V, Hrabač P, Lončar M

PP14 1‐H MRS CHANGES IN DORSOLATERAL PREFRONTAL CORTEX AFTER DONEPEZIL TREATMENT IN PATIENTS WITH MILD TO MODERATE ALZHEIMER'S DISEASE Henigsberg N, Kalember P, Hrabač P, Radoš M, Bajs M, Radoš M, Kovačić Z, Lončar M, Madžar T

PP15 COULD A NEUROLOGICAL DISEASE BE A PART OF MOZART'S PATHOGRAPHY? Ivkić G, Erdeljić V PP16 CORRELATION OF VERBAL MEMORY AND ILLNESS INSIGHT IN THE SCHIZOPHRENIC PATIENTS Janović Š, Bajs M, Đorđević V, Hrabač P, Radonić E, Heningsberg N

PP17 AUTOMATIC SPINDLE DETECTOR FOR INFANT DATA Jerončić A, Achermann P, Đogaš Z

PP18 IS THERE AN IMPACT OF GENETIC MARKERS IN AMYOTROPHIC LATERAL SCLEROSIS PATIENTS ON COGNITIVE IMPAIRMENT? Liščić RM, Muck‐Šeler D, Mustapić M, Babić A, Stukovnik V, Zidar J

PP19 LONG‐TERM MORTALITY IN PERSONS IMPRISONED DURING THE WAR IN CROATIA Lončar M, Henigsberg N, Hrabač P, Gregurek R, Grujić I, Erceg M, Čorić T

PP20 MINIMALLY INVASIVE ANEURYSM TREATMENT AT SCHOOL OF MEDICINE UNIVERSTY OF ZAGREB Mrak G, Paladino J, Radoš M, Jednačak H PP21 HIPPOCAMPAL VOLUME IN SCHIZOAFFECTIVE DISORDER Radonić E, Henigsberg N, Radoš M, Kalember P, Bajs‐Janović M, Folnegović‐Šmalc V PP22 NEUROENDOSKOPSKO LIJEČENJE HIDROCEFALUSA Rotim K, Božić B, Kudelić N, Kčira‐Fideršek V, Saftić R PP23 FUNCTIONAL DIAGNOSTICS OF READING DIFFICULTIES IN DYSPHASIC ADULTS Runjić N, Vranić Đ PP24 CURRENT APPROACHES TO THE PHARMACOTHERAPY OF OPIATE ADDICTION Sakoman S, Jernej B PP25 ANTHROPOMETRIC VARIABLES AND MEAN OXYGEN SATURATION AS PREDICTOR FOR CLINICAL CONSEQUENCES IN OBSTRUCTIVE SLEEP APNEA PATIENTS Ševo V, Selimović M, Vrkić D, Račić G, Đogaš Z


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PP26 THE CHARACTERISTICS OF VISUAL EVOKED POTENTIALS IN SPEECH IMPAIRED CHILDREN Vranić Đ, Runjić N PP27 EEG POLYGRAFIC STUDY OF MATURATIONAL DIFFERENCES BETWEEN TWINS Vučinović M, Ursić A, Rešić B PP28 HYDROXYAPATITE CERAMICS IN MULTILEVEL CERVICAL INTERBODY FUSION ‐ IS THERE A ROLE? Vukić M, Walters CB, Radić A, Mihaljević D, Jednačak H, Marasanov S, Jurjević I

P3 NEUROPHARMACOLOGY PP29 CHANGES OF BRAIN METABOLITES MEASURED WITH MAGNETIC SPECTROSCOPY IN ANTIDEPRESSANT RESPONDERS WITH COMORBID MAJOR DEPRESSION AND POSTTRAUMATIC STRESS DISORDER Henigsberg N, Bajs M, Hrabač P, Kalember P, Radoš M, Radoš M, Radonić E

PP30 PROLONGED EXPOSURE OF RECOMBINANT GABA‐A RECEPTORS TO DIAZEPAM INDUCES DIFFERENTIAL EFFECTS ON FUNCTIONAL COUPLING AND EXPRESSION OF BENZODIAZEPINE BINDING SITES Švob Štrac D, Jazvinšćak Jembrek M, Vlainić J, Peričić D PP31 ANALGESIC OVERUSE IS MORE COMMON IN DEPRESSIVE MIGRAINE PATIENTS Vuković V, Mikula I, Lovrenčić‐Huzjan A, Budišić M, Demarin V PP32 THE EFFECT OF SHORT‐TERM AND LONG‐TERM ZOLPIDEM TREATMENT ON RECOMBINANT α1β2γ2S SUBTYPE OF GABAA RECEPTORS Vlainić J, Jazvinšćak Jembrek M, Peričić D

P4 MOLECULAR NEUROSCIENCE PP33 IMMUNE RESPONSE AND THE MODULATION OF POST‐ISCHEMIC NEUROGENESIS Bohaček I, Gorup D, Lalancette‐Hebert M, Weng YC, Gajović S, Križ J PP34 GLYCOMICS OF BRAIN GLYCOSAMINOGLYCANS BY CHIP‐BASED ELECTROSPRAY IONIZATION ION TRAP MULTISTAGE MASS SPECTROMETRY Flangea C, Sisu E, Seidler D G, Vukelić Ž, Zamfir Alina D

PP35 THE ROLE OF 5HT‐RELATED GENES IN AUTISM: ASSOCIATION STUDY Hranilović D, Novak R, Babić M, Ivanković T, Matak I, Mokrović G, Blažević S, Štefulj J, Jernej B PP36 GANGLIOSIDES OF ADULT HUMAN BRAIN PRECENTRAL AND POSTCENTRAL GYRUS: A CHIP‐MASS SPECTROMETRY BASED COMPOSITION AND STRUCTURE ANALYSIS Jeličić J, Ivković V, Marinčić D, Zamfir Alina D, Vukelić Z

PP37 EXPRESSION ANALYSIS OF STAM2 GENE IN THE BRAIN USING A GENE TRAP DERIVED TRANSGENIC MOUSE MODEL Kapuralin K, Ćurlin M, Dobrivojević M, Srpak N, Mitrečić D, Gajović S PP38 CHARACTERIZATION OF GANGLIOSIDE COMPOSITION OF SELECTED FETAL HUMAN BRAIN REGIONS COMBINING CHIP‐BASED MASS SPECTROMETRY AND THIN‐LAYER CHROMATOGRAPHY Marinčić D, Zamfir Alina D, Kos M, Vukelić Ž

PP39 LOCALIZATION OF DOPAMINE RECEPTORS IN A NEUROBLASTOMA CELL LINE Mladinov M, Diana A, Lam LT, Man NT, Holt I, Morris GE, Šimić G


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PP40 D2, D3 AND D4 DOPAMINE RECEPTOR EXPRESSION IN THE PREFRONTAL CORTEX OF NORMAL AND SCHIZOPHRENIC BRAINS Mladinov M, Mayer D, Jovanov‐Milošević N, Kostović I, Šimić G

PP41 GENE EXPRESSION PROFILING OF CEREBELLAR TISSUE OF GANGLIOSIDE DEFICIENT MICE Mlinac K, Režen T, Heffer M, Rozman D, Kalanj Bognar S PP42 SEROTONIN 5‐HT2A RECEPTOR GENE POLYMORPHISM IN POSTTRAUMATIC STRESS DISORDER Muck‐Šeler D, Babić A, Pivac N, Nedić G, Mustapić M, Kozarić‐Kovačić D

PP43 ASSOCIATION STUDY OF A FUNCTIONAL CATECHOL‐O‐METHYLTRANSFERASE POLYMORPHISM AND COGNITIVE FUNCTION IN PATIENTS WITH DEMENTIA AND MILD COGNITIVE IMPAIRMENT Nedić G, Borovečki F, Klepac N, Mubrin Z, Hajnšek S, Muck‐Šeler D, Pivac N PP44 REPORT ON MUTATION IN EXON 15 OF THE APC GENE IN A CASE OF BRAIN METASTASIS Pećina‐Šlaus N, Majić Ž, Musani V, Zeljko M, Čupić H PP45 AXIN‐1 PROTEIN EXPRESSION AND LOCALIZATION IN GLIOBLASTOMA Pećina‐Šlaus N, Nikuševa Martić T, Kokotović T, Kušec V, Tomas D, Hrašćan R PP46 LIVER AND PANCREAS GLYCOSPHINGOLIPID PHENOTYPES OF THE NEUROLOGICAL DIFFERENT RAT STRAINS Rešić J, Čikeš‐Čulić V, Zemunik T, Markotić A

P5 NEURODEGENERATIVE NEUROSCIENCE PP47 EFFECT OF ENVIRONMENTAL ENRICHMENT ON MORPHOLOGY OF DENTATE GRANULE CELLS AND DEEP LAYER III AND LAYER V PYRAMIDAL CELLS OF OCCIPITAL CORTEX IN OLDEST‐OLD RAT Darmopil S, Petanjek Z, Mohammed Abdul B, Bogdanović N PP48 ASSOCIATIONS OF THE DBH GENE WITH PLASMA DOPAMINE β‐HYDROXYLASE ACTIVITY IN ALZHEIMER'S DISEASE Mustapić M, Presečki P, Mimica N, Pivac N, Folnegović‐Šmalc V, Muck‐ Šeler D PP49 PRELIMINARY STUDY OF THE COGNITIVE FUNCTION AND VAL66MET POLYMORPHISM IN THE BRAIN‐DERIVED NEUROTROPHIC FACTOR GENE IN PATIENTS WITH DEMENTIA AND MILD COGNITIVE IMPAIRMENT Pivac N, Nedić G, Borovečki F, Klepac N, Mubrin Z, Hajnšek S, Nikolac M, Muck‐Šeler D

PP50 OLIVE‐OIL ENRICHED DIET AND CHANGES IN TOTAL LIPIDS CONTENT OF BRAIN FROM MICE IN THE EARLY PHASE OF LIVER REGENERATION Pantović R, Milin Č PP51 ENVIROMENTAL ENRICHMENT HAS BENEFICIAL EFFECT ON COGNITION IN THE RAT MODEL OF SPORADIC ALZHEIMER’S DISEASE Osmanović J, Šalković‐Petrišić M, Riederer P


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STUDENT ORAL PRESENTATIONS Saturday, September 26th 2009

11.30 ‐12.30 OP1 INFLUENCE OF SHAPE ON LIKENESS OF OBJECTS Ugrina Marija Eva University of Zadar, Department of Psychology OP2 GENE EXPRESSION PROFILING OF CEREBELLAR TISSUE OF GANGLIOSIDE DEFICIENT MICE Mlinac Kristina

Croatian Institute for Brain Research, School of Medicine, University of Zagreb OP3 CHARACTERISTICS OF GLIAL CELLS OF THE SUBPLATE ZONE IN DIFFUSE AND FOCAL PERIVENTRICULAR WHITE MATTER INJURY Ivana Pogledić Croatian Institute for Brain Research, School of Medicine, University of Zagreb OP4 GENDER AND AGE RELATED DIFFERENCES IN RECOGNITION OF FACIAL EXPRESSIONS OF EMOTIONS Barbir L, Gregorić B, Harhaj A, Ilakovac V, Heffer‐Lauc M School of Medicine, University of Osijek OP5 STIMULATION OF FUNCTIONAL VISION IN CHILDREN WITH PERINATAL BRAIN DAMAGE Alimović S, Katušić A, Mejaški‐Bošnjak V

OP6 EFFECTS OF VIBROTACTILE STIMULATION ON THE CONTROL OF SPASTICITY AND MOVEMENTS FACILITATION IN CHILDREN WITH CEREBRAL INJURY Katušić A , Alimović S , Mejaški‐Bošnjak V


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POSTER WALK II

Saturday, September 26th 2009 12.30 ‐ 14.00

P6 COGNITIVE NEUROSCIENCE PP52 EARLY PHONOLOGICAL DEVELOPMENT AT CHILDREN WITH EARLY BRAIN INJURY Blaži D, Kovač‐Gornji N PP53 GAZE DIFFERENCES IN PROCESSING PLEASANT AND UNPLEASANT COMPLEMENTARY PICTURES Budimir S, Palmović M

PP54 EARLY COMMUNICATION DEVELOPMENT OF SOCIALLY DEPRIVED CHILDREN: ARE THERE SIGNS OF "INSTITUTIONAL AUTISM"? Cepanec M, Gmajnić I, Ljubešić M PP55 COMPARISON OF EMOTION RECOGNITION IN FACIAL EXPRESSION AND MUSIC Gašpar T, Jurić Iva, Labor M, Dumančić D, Ilakovac V, Heffer‐Lauc M PP56 COGNITIVE ABILITIES AND LANGUAGE COMPREHENSION IN PRESCHOOL CHILDREN WITH PERINATAL BRAIN LESION Ivšac Pavliša J, Šimleša S, Ljubešić M PP57 SPATIAL AND TEMPORAL MEASUREMENTS OF EYE MOVEMENT IN CHILDREN WITH DYSLEXIA: INDIVIDUAL DIFFERENCES Kuvač Kraljević J, Palmović M PP58 EFFECTS OF PRESENTATION TIME AND INTERPOLATED ACTIVITY ON FALSE MEMORY Manenica I, Prenđa S PP59 APPLICATION OF THE CONTROLLED ORAL WORD ASSOCIATION TEST IN A CROATIAN SAMPLE Mimica N, Žakić Milas D, Joka S, Kalinić D, Folnegović Šmalc V, Harrison John E

PP60 KOGNITIVNI SLUŠNI EVOCIRANI POTENCIJALI U DJECE S GOVORNO‐JEZIČNIM POTEŠKOĆAMA Munivrana B, Išgum V, Orlović J, Marn B PP61 POSTADOLESCENT CIRCUITRY REORGANIZATION OF ASSOCIATIVE LAYER IIIC PYRAMIDAL NEURONS IN THE HUMAN PREFRONTAL CORTEX Petanjek Z, Zeba M, Uylings BM H, Kostović I PP62 EFFICACY IN FITT'S TAPPING TASKS ACROSS MENSTRUAL CYCLE Šimić N, Lovrić M, Šuto A PP63 EFFICIENCY IN SPATIAL AND VERBAL TASKS DURING THE MENSTRUAL CYCLE Šimić N, Santini M PP64 USEFULNESS OF DRAWING AND WRITING TEST WITH BOTH HANDS IN SUBJECTIVE AND OBJECTIVE DETERMINATION OF DOMINANT HEMISPHERE Šnajder D, Labak I, Kostović‐Srzentić M, Benčić M, Ništ M, Ilakovac V, Heffer‐Lauc M PP65 EYSENCK’S TEMPERAMENT TYPOLOGY AND EVOKED POTENTIALS Tatalović Vorkapić S, Tadinac M


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P7 BASIC NEUROSCIENCE

PP66 EXPRESSION OF SEROTONIN TRANSPORTER IN PERIPHERAL BLOOD LYMPHOCYTES FROM PATIENTS WITH ALCOHOL DEPENDENCE Antica M, Matošić A, Marušić S, Jernej B, Čičin‐Šain L

PP67 EXPRESSION AND FUNCTION OF 5HT‐1A RECEPTOR IN BRAINS OF WISTAR‐ZAGREB 5HT RATS Bordukalo‐Nikšić T, Čičin‐Šain L, Jernej B PP68 PROPOFOL ATTENUATES THE PHRENIC LONG TERM FACILITATION IN RATS SUBJECTED TO ACUTE INTERMITTENT HYPOXIA Carev M, Valić M, Pecotić R, Karanović N, Valić Z, Pavlinac I and Đogaš Z

PP69 RhoB PROTEIN IS INVOLVED IN CELL RESPONSE TO GENOTOXIC STRESS Cimbora‐Zovko T, Fritz G, Osmak M PP70 PRENATAL DEVELOPMENT OF NITRINERGIC NEURONS IN THE HUMAN BASAL FOREBRAIN Džaja D, Ažman D, Pletikos M, Judaš M PP71 SINGLE ELECTROCONVULSIVE SHOCK DOES NOT INFLUENCE COX‐2 AND HSP70 EXPRESSIONS IN THE RAT FRONTAL CORTEX AND HIPPOCAMPUS Hrelja A, Pilipović K, Župan G PP72 HUMAN FOETAL SPERMATOGONIA EXPRESS NEURON‐SPECIFIC ENOLASE Ježek D, Šklebar D, Kozina V, Šklebar I, Kos M PP73 TRANSIENT MAP2‐POSITIVE CELLULAR BELT IN THE CORTICAL PLATE COINCIDE WITH BEGINNING OF THE AREAL SPECIFICATION. Jovanov‐Milošević N, Petrović D, Judaš M, Kostović I PP74 SPATIAL LEARNING AND MEMORY IN WISTAR‐ZAGREB 5HT RAT Mokrović G, Jernej B, Čičin‐Šain L PP75 QUALITATIVE AND QUANTITATIVE PROPERTIES OF DENDRITIC MORPHOLOGY COMPARING DIFFERENT SUBPOPULATIONS OF PRINCIPAL NEURONS IN INFRAGRANULAR LAYERS OF THE HUMAN PREFRONTAL CORTEX Paladin I, Petanjek Z PP76 BLOCKADE OF 5‐HT1A RECEPTORS IN THE RAT PHRENIC NUCLEUS ATTENUATED RAPHE INDUCED ACTIVATION OF THE PHRENIC NERVE ACTIVITY Pecotić R, Đogaš Z, Valić Z, Valić M

PP77 DEVELOPMENT OF MICROGLIA IN THE RHESUS MONKEY DURING MIDGESTATION Petanjek Z, Džaja D, Milašin G PP78 ORIGINS AND MIGRATORY ROUTES OF GABA‐ERGIC NEURONS IN THE PRIMATE CEREBRAL CORTEX Petanjek Z, Kostović I, Hladnik A, Esclapez M PP79 CHANGES OF OXIDATIVE STRESS PARAMETERS IN DIFFERENT RAT BRAIN REGIONS FOLLOWING TRAUMATIC BRAIN INJURY Pilipović K, Župan Ž, Frković V, Dangubić B, Mršić‐Pelčić J, Šustić A, Župan G

PP80 SERUM LIPID LEVELS IN ALZHEIMER'S DISEASE Presečki P, Muck‐Šeler D, Mimica N, Pivac N, Mustapić M, Folnegović‐Šmalc V

PP81 SEROTONIN (5HT) METABOLISM IN THE BRAIN OF WISTAR‐ZAGREB 5HT RATS Štefulj J, Čičin‐Šain L, Bokulić Z, Mokrović G, Bordukalo‐Nikšić T, Orešković D, Živin M, Jernej B


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PP82 DIFFERENT EXPERIMENTAL PROTOCOLS AND PHRENIC LONG‐TERM FACILITATION Valić M, Pecotić R, Pavlinac I, Valić Z, Đogaš Z

PP83 DYNAMICS OF THE VOLUME CHANGES OF THE TRANSIENT CELLULAR ZONES IN THE DEVELOPING HUMAN CEREBRAL WALL Vasung L, Evans A, Kostović I

PP84 DISTRIBUTION OF MAJOR BRAIN GANGLIOSIDES IN OLFACTORY TRACT OF FROGS Viljetić B, Degmečić IV, Krajina V , Bogdanović T, Mojsović A, Đikić D, Vajn K, Heffer‐Lauc M

PP85 DENDRITIC REORGANIZATION OF GRANULE CELLS IN THE FASCIA DENTATA OF THE THY1‐GFP MOUSE FOLLOWING ENTORHINAL CORTEX LESION Vukšić M, Del Turco D, Muller Christian M, Deller T

P8 NEURODEVELOPMENTAL BASIS OF COGNITIVE, MENTAL AND NEUROLOGICAL DISORDERS PP86 LENTICULOSTRIATAL VASCULOPATHY (LSV) ‐ A MARKER FOR CONGENITAL CMV INFECTION? Đuranović V, Mejaški‐Bošnjak V, Lujić L, Krakar G, Gojmerac T PP87 CHARACTERISTICS OF HEARING LOSS IN UNILATERAL CLEFT LIP AND PALATE – INFLUENCE ON COMMUNICATION Handžić J, Vladika I, Šimunčić Veselić A, Bura M, Radić B PP88 INFANTILE SPASMS IN CHILDREN WITH DOWN SYNDROME Lujić L, Mejaški Bošnjak V, Delin S, Đuranović V, Krakar G PP89 BRAIN MALFORMATIONS IN CHILDREN WITH CONGENITAL CYTOMEGALOVIRUS Mejaški‐Bošnjak V, Krakar G, Đuranović V, Rakvin I, Delin S PP90 EARLY INTERVENTION IN INFANTS AT HIGH RISK FOR A NEURODEVELOPMENTAL DISORDERS Pinjatela R, Joković Oreb I

P9 SPECIAL SESSION – CASE REPORTS

PP91 DIFFERENTIAL DIAGNOSTIC RELEVANCE OF HIGH RESOLUTION MAGNETIC RESONANCE IN PATIENTS WITH PROBABLE EARLY MULTIPLE SYSTEM ATROPHY (MSA) – A CASE REPORT Bačić Baronica K, Ivkić G, Ozretić D PP92 DEPRESSIVE EPIZODE IN AN ADOLESCENT GIRL WITH TUBEROSE SCLEROSIS COMPLEX Boričević Maršanić V. PP93 CEREBELLAR GLIOBLASTOMA IN ELDERY PERSON ‐ CASE REPORT Božić B, Rotim K, Kčira‐Fideršek V, Krpina H, Čupić H PP94 MAGNETIC RESONANCE FINDINGS IN A NEONATE WITH NONKETOTIC HYPERGLYCINEMIA ‐ CASE REPORT Čuljat M, Benjak V, Dasović‐Buljević A, Ozretić D, Fumić K, Barić I PP95 ACUTE LONGITUDINALLY EXTENSIVE TRANSVERSE MYELITIS IN SIX YEAR OLD CHILD‐CASE REPORT Delin S, K Pavešić K,Tešović G, Knezović I, Miše B


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PP96 APPLICATION OF NEUROPHYSIOLOGIC BACKGROUND OF NORMAL MOVEMENT IN PHYSIOTHERAPIST TREATMENT WITH BOBATH CONCEPT BY OPTIMIZING THE FUNCTION OF UPSTANDING ACITIVITIES IN ADULT PATIENTS WITH ACQUIRED LESIONS OF THE CNS ‐ CASE REPORT Jelica S, Seper V, Davidović E PP97 TREMOR, SEIZURES, PSYCHOSIS AND DIPLOPIAE IN LYME‐NEUROBORRELIOSIS: REPORTS OF TWO CASES Šarac H, Markeljević J

PP98 DURAL, JUVENILE AND PERIMEDULLARY ARTERIOVENOUS SHUNTS: REPORT OF FOUR CASES Šarac H, Telarović S, Vranješ D, Žagar M, Markeljević J, Ozretić D

PP99 AGGRESSIVE INTESTINAL SCHWANNOMA MALIGNUM MIMICKING GYNECOLOGICAL PATHOLOGY Tomica D, Danolić D, Puljiz M, Alvir I, Mamić I, Milas I, Banović M


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BOOK OF ABSTRACTS


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PRESIDENTIAL SYMPOSIUM

“NEW ADVANCES IN NEUROSCIENCE”


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GENOMIC LANDSCAPE OF AUTISM SPECTRUM DISORDER Maja Bućan University of Pennsylvania, Philadelphia, USA Autism spectrum disorders (ASDs) represent a group of childhood neurodevelopmental and neuropsychiatric disorders characterized by deficits in verbal communication, impairment of social interaction, and restricted and repetitive patterns of interests and behaviour. To identify common genetic risk factors underlying ASDs, we performed genome‐wide association studies on a cohort of 940 families with affected children and 2000 control subjects. Six single nucleotide polymorphisms between cadherin 10 (CDH10) and cadherin 9 (CDH9)‐two genes encoding neuronal cell‐adhesion molecules‐revealed strong association signals (Wang et al., Nature 459, 2009). To identify copy number variations (CNVs) that are associated with an increased risk of ASDs we selected more than 150 loci harboring rare variants in multiple unrelated probands, but no controls. Rare variants at known loci, including exonic deletions at NRXN1 and whole gene duplications encompassing UBE3A and several other genes in the 15q11‐q13 region, were observed in the course of these analyses. Strong support was likewise observed for previously unreported genes such as BZRAP1, an adaptor molecule known to regulate synaptic transmission, with eDels or eDups observed in twelve unrelated cases but no controls. Less is known about MDGA2, likewise observed to be case‐specific. But, it is notable that the encoded protein shows an unexpectedly high similarity to Contactin 4, which has also been linked to disease. That hundreds of distinct rare variants were each seen only once further highlights complexity in the ASDs and points to the continued need for larger cohorts (Glessner et al., Nature 459, 2009; Bucan et al., PLoS Genetics e1000536). Finally, I will present results of a pilot study on the genetic analysis of 100 ASD subjects recruited in several centers for Autism across Croatia (collaboration with Drs. D. Hranilovic, Z. Bujas‐Petkovic and B. Jernej).


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THE USE OF TRYPTOPHAN ANALOG IN STUDYING THE BRAIN SEROTONERGIC SYSTEM Mirko Dikšić Department of Neurology and Neurosurgery, and Montreal Neurological Institute, McGill University, Montreal, Canada and Faculty of Medicine, J.J. Strossmayer University, Osijek, Croatia The study of regional serotonin synthesis in humans by imaging could enable researchers to obtain a better understanding of affective disorders and to develop better therapies. Studies were performed on laboratory animals using two different rat models of depression, where the synthesis was measured by autoradiography, and a 14C and 3H‐labelled tracer. Positron emission tomography (PET) with a 11C‐labelled α-methyl‐L‐tryptophan [α-MTrp] tracer was used in the human studies, on both normal subjects and on patients with various brain disorders (e.g., epilepsy, depression, obsessive compulsive disorder, borderline personality disorder, and those suffering from migraines). The experiments in rats have shown that 5‐HT synthesis is elevated in bulbectomized rats (using a model of agitated depression) and reduced in the Flinders Sensitive Line rats (using a model of retarded depression), and that antidepressants (e.g., citalopram, buspirone) have the effect of returning the synthesis to the level of the control rats without having a significant effect on plasma Trp concentration. In healthy women, when compared to healthy men, serotonin synthesis is significantly lower in the right parietal lobe, bilateral middle frontal gyri, and bilateral parieto‐occipital lobe. In depressed patients, significant bilateral decreases in the anterior cingulate (ACC) (females), in the left ACC (males), and in the left mesial cortex (both gender) were found. In addition, we have shown that antidepressants have region specific influence on 5‐HT synthesis. In rat models of depression was shown that antidepressant produces changes in 5‐HT synthesis and some 5‐HT receptor sites. Some of these correlate with behavioural changes.


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EXTRACELLULAR PROTEOLYSIS CONTROLLING SYNAPTIC PLASTICITY Leszek Kaczmarek Laboratory for Molecular Neurobiology, Nencki Institute of Experimental Biology, Warsaw, Poland http://neurogene.nencki.gov.pl Matrix metalloproteinase 9 (MMP‐9) is an extracellularly operating enzyme that has recently been implicated in dendritic remodeling, synaptic plasticity, learning and memory (Szklarczyk et al., J. Neurosci., 2002; Nagy et al., J. Neurosci., 2006; Meighan et al., J. Neurochem., 2006; Okulski et al., Biol. Psych., 2007). Furthermore, we have recently identified MMP‐9 as a being produced, expressed and active at the synaptic contacts (Konopacki et al., Neuroscience, 2007; Michaluk et al., J. Biol. Chem., 2007; Wilczynski et al., J. Cell Biol., 2009). Furthermore, MMP‐9 has been identified as a key pathogenic factor in two animal models of temporal lobe epilepsy which is a devastating disease in which aberrant synaptic plasticity plays a major role (Wilczynski et al., 2008). We have also shown that MMP‐9 upon excitatory stimulation can cleave beta‐dystroglycan (Michaluk et al., J. Biol. Chem., 2007) the transmembrane protein anchoring dystrophin complex in the cell membrane, which via actinin can interact with membrane receptors for glutamate, the major excitatory neurotransmitter in the brain. Recently, using single molecule tracking we have found that enzymatic activity of MMP‐9 increases lateral dendritic mobility of specific class of glutamate receptors, namely NMDA receptors (Michaluk et al., J. Neurosci., 2009). On the other hand, MMP‐9 has not exerted any effects on the mobility of another kind on glutamate receptors, i.e, the AMPA ones. The mechanism of MMP‐9 action on NMDAR is caused neither by change in overall ECM structure, nor by cleavage of NMDAR subunits, however it does depend on integrin beta 1. Our most recent results suggest that MMP‐9 may directly influence morphological reorganization of dendritic spines that are postsynaptic domains believed to serve as the major substrate for neuronal plasticity.

http://neurogene.nencki.gov.pl/


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MOLECULAR CONTROL OF NEURONAL IDENTITY AND CONNECTIVITY IN THE CEREBRAL CORTEX Nenad Šestan Department of Neurobiology, Kavli Institute for Neuroscience, Yale University School of Medicine, New Haven Certain traits, such as the arrangement of neurons in the cerebral cortex into layers containing unique sets of cell types and synaptic connections, seem to be conserved across mammals. We are interested in defining the molecular mechanisms that underlie the astonishing diversity of layer‐ and region‐specific cell types and axonal projections in the mouse. In doing so, we hope to uncover the evolutionarily conserved core genetic programs that regulate molecular identities and axonal projections of cortical neurons. The local differences in the laminar pattern of cortical projection (pyramidal) neurons and their connections underlie the separation of the cortex into anatomically and functionally distinct areas. For example, layer 5 pyramidal neurons throughout the neocortex project to subcortical regions. However, the layer 5 neurons of the frontal lobe project to the motor nuclei in the brain stem and spinal cord, while those in the visual cortex project only to the tectum. Also, pyramidal neurons arising from the same lineage acquire distinct layer‐specific identities based on the timing of their birth during neurogenesis. A neuron’s laminar identity is intimately linked to its eventual function; pyramidal neurons of the deepest layers (6 and 5) send subcortical axonal projections to the thalamus and brainstem/spinal cord, respectively, while those of the upper layers (2‐4) exclusively form intracortical connections. How layer‐ and area‐specific genetic programs arise and translate into functional differences among pyramidal neurons is the main focus of this presentation. Ongoing studies are aimed at identifying genetic and molecular mechanisms involved in specifying the identities of pyramidal neurons, the patterning of their axonal projections, and the formation of synaptic connections of the cerebral cortex.


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MINI SYMPOSIUM

„APPROACHES IN TRANSLATIONAL NEUROSCIENCE“


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WISTAR‐ZAGREB 5HT RATS: FROM PLATELETS TO BRAIN Jernej Branimir, Čičin‐Šain Lipa Laboratory of Neurochemistry and Molecular Neurobiology, Ruđer Bošković Institute, Bijenička 54, 10000 Zagreb, Croatia By selective breeding for extreme values of platelet serotonin level (PSL), two sublines of rats with constitutional hyperserotonemia/hyposerotonemia were developed. The velocity of platelet serotonin uptake (PSU), the main determinant of PSL, was used as further, more specific, selection criterion. Directed breeding for its extremes resulted in two sublines of rats with constitutional up‐regulation/down‐regulation of platelet 5HT transporter activity, and showed consequent alterations of entire 5HT homeostasis. These sublines, termed Wistar‐Zagreb 5HT rats (WZ‐5HT rats), constitute a genetic rodent model which will be presented. Besides changes in peripheral 5HT homeostasis, high‐5HT and low‐5HT sublines of WZ‐5HT rats also demonstrate changes in central serotonergic mechanisms. Under physiological conditions, neurochemical differences in the 5HT system between sublines were almost undetactable, but they became evident upon specific pharmacologic challenge, as shown by brain microdialysis study. Differential behavioral phenotypes of 5HT sublines in response to various environmental challenges provide further evidence for differences between sublines of WZ‐5HT rats and strongly indicate the brain serotonergic activity is increased in animals from the high‐5HT subline as compared to low‐5HT rats. The WZ‐5HT rat model may represent an integrative model for serotonin and serotonin transporter research, incorporating changes at genomic/genetic and phenotypic (neurodevelopmental, structural, biochemical, behavioral etc.) levels and encompassing both central and peripheral 5HT functioning.

‐ RECENT FINDINGS IN ETIOPATHOGENESIS OF ALZHEIMER'S DISEASE ‐ DOES ALZHEIMER’S DISEASE BEGIN IN THE DORSAL RAPHE NUCLEUS? Šimić Goran Croatian Institute for Brain Research, School of Medicine University of Zagreb, Šalata 12, 10000 Zagreb, Croatia Although substantial evidence indicates that the progression of pathological changes of the neuronal cytoskeleton is crucial in determining the severity of dementia in Alzheimer’s disease (AD), the exact causes and evolution of these changes, the initial site at which they begin, and the neuronal susceptibility levels for their development are poorly understood. The current clinical criteria for diagnosis of AD are focused mostly on cognitive deficits produced by dysfunction of hippocampal and high‐order neocortical areas, whereas noncognitive, behavioural and psychological symptoms of dementia such as disturbances in mood, emotion, appetite, and wake–sleep cycle, confusion, agitation and depression have been less considered. The early occurrence of these symptoms suggests brainstem involvement, and more specifically of the serotonergic nuclei. In spite of the fact that the Braak and Braak staging system and National Institutes of Aging – Reagan Institute (NIA‐RI) criteria do not include their evaluation, several recent reports drew attention to the


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possibility of selective and early involvement of raphe nuclei, particularly the dorsal raphe nucleus (DRN), in the pathogenesis of AD. Based on these findings of differential susceptibility and anatomical connectivity, a novel pathogenetic scheme of AD progression was proposed. Although the precise mechanisms of neurofibrillary degeneration still await elucidation, we speculated that cumulative oxidative damage may be the main cause of DRN alterations, as the age is the main risk factor for sporadic AD. Within such a framework, β‐amyloid production is considered only as one of the factors (although a significant one in familial cases) that promotes molecular series of events underlying AD‐related neuropathological changes. NOVEL FINDINGS IN STREPTOZOTOCIN MODEL OF SPORADIC ALZHEIMER’S DISEASE: THE ORIGIN OF SPORADIC ALZHEIMER’S DISEASE – IS IT BRAIN INSULIN RESISTANT STATE? Šalković‐Petrišić Melita1, Osmanović Jelena1, Grünblat Edna2, Hoyer Siegfried3, Riederer Peter2 1Department of Pharmacology and Croatian Institute for Brain Research, Medical School, University of Zagreb, Zagreb, Croatia, 2Department of Clinical Neurochemistry, Clinic for Psychiatry and Psychotherapy, Bayerische Julius‐Maximilians‐ University of Würzburg, Würzburg, Germany, 3Department of Pathology, University of Heidelberg, Heidelberg, Germany. Sporadic Alzheimer disease (sAD) is associated with brain insulin receptor (IR) signalling abnormalities whose ethiopathogenesis in the course of sAD is still unclear. Rats that have been intracerebroventricularly treated with streptozotocin (STZ‐icv), a drug selectively toxic for insulin producing/secreting cells and IR, have been recognized recently as the non‐transgenic, experimental sAD model, suitable for exploration of brain IR signalling cascade dysfunction. We investigated the time course of brain IR signalling dysfunction in relation to the amyloid pathology development in STZ‐icv rat model of sAD. RT‐PCR and immunoblotting were used for hippocampal IR signalling cascade gene and protein expression measurements, respectively and data were analyzed by Cruscal‐Wallis ANOVA median/Mann‐Whitney U test. Amyloid β (Aβ) aggregation was detected by Congo red staining and immunohistochemistry. Our results demonstrated alterations in brain IR signalling cascade starting from the decreased insulin gene expression, followed by a decreased expression of IR mRNA and protein, found as early as 1 month following the induction of sAD‐like condition and persisting up to 9 months afterwards. Downstream dysfunction of IR signalling pathway manifested as decreased ratio of phosphorylated/non‐phosphorylated glycogen synthase kinase‐3 was found not earlier than 3 months after STZ‐icv treatment and was followed by tau protein hyperphosphorylation, cerebral amyloid angiopathy, intraneuronal Aβ aggregation and plaque‐like formation (not earlier than 6 months after the STZ‐icv treatment). No change in amyloid precursor protein gene expression was found while the expression of insulin degrading enzyme gene/protein was decreased. Based on this experimental approach, sAD could be considered as an insulin resistant brain state which precedes and eventually triggers Aβ pathology. Supported by DAAD and MZOS.


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PATHOPHYSIOLOGY OF INTRACRANIAL HYPERTENSION Klarica Marijan, Radoš Milan, Vukić Miroslav2, Orešković Darko1, Bulat Marin Department of Pharmacology and Croatian Institute for Brain Research, School of Medicine University of Zagreb, Šalata 11, Zagreb, Croatia 1Ruđer Bošković Institute, Bijenička 54, Zagreb, Croatia 2Department of Neurosurgery, School of Medicine University of Zagreb, Šalata 3, Zagreb, Croatia

Intracranial hypertension and neurotrauma are leading causes of death in a population up to 40 years of age in developed countries. Pathophysiology of the intracranial hypertension caused by neurotrauma and other pathological conditions (intracranial bleeding, brain tumors, brain edema, meningitis, encephalitis, hydocephalus, etc.) is still unclear. Therefore, the treatment of intracranial hypertension is a difficult problem with an uncertain outcome. It is generally accepted that the intracranial pressure is regulated by interaction of three main intracranial volumes (volume of brain, cerebrospnial fluid‐CSF, and cerebral blood). It is also believed that CSF volume is a result of the ratio between the rate of secretion and absorption inside the cranium, and that this is one of the main factors in regulation of intracranial pressure. We have found that increase of cranial CSF pressure is observed only when spinal intradural volume is filled up, and that increase or decrease of spinal CSF volume causes augmentation or fall of cranial CSF pressure. Thus, it appears that intracranial CSF pressure depends on total CSF volume in craniospinal space and that spinal CSF space is the main regulatory factor in pathophysiology of intracranial hypertension. It would appear that the volume of spinal CSF in men is significantly larger than previously assumed. According to the recent research, it appears that the secretion, circulation and absorption of CSF do not exist, but the CSF volume is regulated by the osmotic and hydrostatic pressures that exist between the CSF, interstitial fluid and cerebral capillaries. In further researches it will be necessary to determine the role of osmotic and hydrostatic forces in regulation of CSF volume. In addition, it is important to measure the total CSF volume in craniospinal space under normal pressure, and changes of CSF volume in cranial and spinal space during the increase and decrease of CSF pressure. Better understanding of the intracranial pressure regulation and CSF physiology should result in new and more efficient approach in treatment of intracranial hypertension.


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POSTER WALK I

P1 CEREBROSPINAL PATHOPHYSIOLOGY AND APPLICATION OF ULTRASOUND PP01 THE USAGE OF ULTRASONIC PROBE "NECUP‐2" IN NEUROENDOSCOPIC PROCEDURES Jednačak H1, Paladino J1, Miklić P1, Vukić M1, Štimac T2, Klarica M3

1Department of Neurosurgery, School of Medicine, University of Zagreb, 2Brodarski Institute, Zagreb, 3Center for Clinical Research in Neuroscience and Croatian Institute for Brain Research, Zagreb, Croatia Introduction: The number of endoscopic procedures in neurosurgery is rapidly growing, not only in treatment of different obstructive hydrocephalus forms, but also in other indications: various intracranial cysts and intraventricular space occupying lesions. The usage of different neuroendosopic techniques and instruments (blunt perforation, endoscopic scissors and forceps, mono and bipolar electrocoagulation and laser) clearly demonstrates unsufficiency of standard neuroendoscopic armamentarium. We report our experience with originally constructed Neurosurgical Endoscope Contact Ultrasound Probe "NECUP‐2" in neuroendoscopy. Material and Methods: The newly developed "NECUP‐2" consists of the ultrasound generator (amplitude 300 ?m, intensity 900 W/cm2, acceleration 400000 m/s2, frequency 25000 Hz), and titanium microprobe with 1.6 mm diameter which easily passes through the 2.2 mm ventriculoscope working channel. The ultrasound intensity can be changed from low to high, enabling tissue perforation and removal with preserving of the vascular structures. Results and Conclusion: Between June 1997 and June 2007, 132 neuroendoscopic procedures have been performed: 102 ETV, 10 septotomies, 15 arachnoid cysts and 5 intraventricular tumors. The "NECUP‐2" was applied effectively in all cases in which blunt perofration was not possible: 38/102 ETV, 10/10 septostomies, 15/15 arachnoid cysts. In the five cases of intraventricular tumors neuroendosopic procedure was combined with open microsurgery for tumor removal with preservation of vascular structures. There were no "NECUP‐2" related complications. Our results demonstrate that the application of the "NECUP‐2" in neurosurgical endoscopic procedures is safe and effective, and can be a useful addition to standard neuroendoscopic instruments and techniques. PP02 PHYSICAL CHARACTERISTICS IN THE NEW MODEL OF THE CEREBROSPINAL FLUID SYSTEM Jurjević I, Radoš M, Orešković J, Prijić R, Klarica M Department of Pharmacology, School of Medicine University of Zagreb and Croatian Institute for Brain Research, Zagreb, Croatia Introduction: It is unknown which factors determine the changes in cerebrospinal fluid (CSF) pressure inside the craniospinal system during the changes of the body position. To test this, we developed a new model of the CSF system, which by it's biophysical characteristics and dimensions imitates the CSF system in cats. The results obtained on a model were compared to those in animals observed during changes of body position. Methods and Results: A new model was constructed from two parts with different physical characteristics. The „cranial" part is developed from a plastic tube with unchangeable volume, while the „spinal" part is made of a rubber baloon, with modulus of elasticity similar to that of animal spinal dura. In upright position, in the „cranial" part of the model the negative pressure appears without any measurable changes in the fluid volume, while in „spinal" part the fluid pressure is positive and consistent with the hydrostatic pressure . All of the observed changes are in accordance to the fluid mechanics. Alterations of the CSF pressure in cats during the changes of the body position are not significantly different compared to those observed on our new model.


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Conclusion: This suggests that the CSF pressure changes are related to the fluid mechanics, and do not depend on CSF secretion and circulation. It seems that in all body positions the cranial volume of blood and CSF remains constant, which enables a good blood brain perfusion.

PP03 POTENTIAL ERROR IN VENTRICULO‐CISTERNAL PERFUSION METHOD FOR DETERMINATION OF CEREBROSPINAL FLUID RATE IN CATS Orešković D1, Maraković J2, Radoš M3, Jurjević I3, Klarica M3

1Ruđer Bošković Institute, Zagreb, 2Dubrava University Hospital, Zagreb, 3Department of Pharmacology, School of Medicine University of Zagreb and Croatian Institute for Brain Research, Zagreb, Croatia Introduction: The cerebrospinal fluid (CSF) formation rate (Vf) has been extensively studied by the ventriculo‐cisternal perfusion technique, which is still regarded as the most precise method. This method and the equation for the calculation of the CSF formation rate have been established by Heisey et al, who assumed that the dilution of the indicator substance is a consequence of newly formed CSF, i. e. that a higher CSF formation rate will result in a higher degree of dilution of the indicator substance. Therefore, this method is indirect and any mistake in the interpretation of the degree of dilution of the indicator substance in the perfusate caused by other reasons would result in questionable and often contradictory conclusions regarding CSF formation rates. Materials and Methods: In chloralose anaesthetised cats, ventriculo‐cisternal perfusion was performed at higher (252 |iL/min) and at lower perfusion rate (65.5 |iL/min) and Vf was calculated by equation Vf = Vi x (Ci ‐ Co)/Co, where Vi represents inflow rate; Ci concentration of indicator substance in inflow perfusate and Co represents concentration of indicator substance in outflow perfusate. During perfusion, the indicator substance is partially absorbed into surrounding tissue (10% of Co) and therefore corrections of experimental results obtained on cats for this absorption of the indicator substance have been made. Results and Conclusion:Under previously described experimental conditions we have obtained higher corrected results (27.9 |iL/min) at higher rate of perfusion (252 |iL/min) than those (9.0 |iL/min) at lower rate of perfusion (65.5 |iL/min). Since corrected difference of 19.9 |iL/min (27.9 ‐ 9.0) of CSF formation rate should not have occurred, and considering the fact that, beside indicator substances, water is absorbed from the perfusate into surrounding tissue as well, obtained results have clearly shown that ventriculo‐cisternal perfusion method cannot be used as reliable method for measuring CSF formation rate. PP04 DEVELOPMENT OF NEW ULTRASONIC PROBES FOR NEUROSURGICAL AND MICROSURGICAL PROCEDURES Paladino J1, Štimac T2, Svilar D2, Klarica M3

1Department of Neurosurgery, KBC Rebro, School of Medicine University of Zagreb, 2Brodarski Institute, Zagreb, 3Center for Clinical Research in Neuroscience, School of Medicine, Zagreb, Croatia During the last sixteen years the cooperation on projects of application of high power ultrasound on the neural and other tissues, is established between experts from The Department of Neurosurgery KBC Rebro, School of Medicine, The Brodarski Institute, and The Center for Clinical Research in Neuroscience, School of Medicine, Zagreb. As a result of these projects three generations of ultrasonic devices, which can be used in microsurgery, have been developed. The first device called NECUP‐1 (Neurosurgical Endoscope Contact Ultrasound Probe) has been constructed to improve the procedure of endoscopic ventriculocysternostomy. This was the first time that the ultrasonic wired contact probe was used in the treatment of hydrocephalus, taking place of so far used electric monopolar and bipolar coagulators. The operation was performed with ventriculoscope Aesculap FF 370, <D= 6.2 mm, with a newly developed ultrasonic probe <D= 1.6 mm, controlled by a camera. An elastic titanium wire invoked with a high energy ultrasound of approximately 25 kHz frequency was administered in one of the working openings. The tip of the probe was directed to the tissue which had to be removed due to several ultrasonic effects (strong


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longitudinal vibrations; cavitation; "hammer" effect). Upon completion of the first model we started the development of a new device called NECUP‐2 which was then successfully applied in clinical practice in year 2004. That device is composed of ultrasonic generator, ultrasonic probe and additional special instruments made of titanium. During conception and development of device, the computer aided system was used, so special emphasis was placed on manufacturing of instruments which were adjusted for different types of surgical procedures (specific shape of scalpels, cutters and cleaners, etc). In the end of 2008., the third generation of the device was finished (UPROMS‐1). Six different kinds of ultrasonic probes can be connected to the universal ultrasonic generator, using the ultrasonic concentrators of various shapes (cone, cylindrical, exponential etc.). The probes can be adjusted for different applications (neurosurgery, surgery of the liver, urology, cardio surgery etc.). In this way fully developed and adjusted in frequency, acoustic transformers (concentrators) enable that the additional carrying‐out instruments can be connected to them. The specificity of the UPROMS‐1 device is that a wire of different length (10‐90 cm; ~ nX/2 ~ 25 kHz) and diameter (<D=1.5, 1.6 and 2 mm) can be used for endoscopic application. PP05 METHODS FOR MEASURING ACOUSTIC POWER OF ULTRASONIC NEUROSURGICAL DEVICE Petošić A1, Ivančević B1, Svilar D2, Štimac T2, Klarica M3

1Faculty of Electrical Engineering and Computing, Zagreb, Unska 3, Croatia, 2Brodarski Institute, Zagreb, Avenija Većeslava Holjevca bb, Croatia, 3Center for Clinical Research in Neuroscience and Croatian Institute for Brain Research, Zagreb, Croatia Introduction: Measurement of the acoustic power radiated by the high‐energy ultrasonic devices is complex due to occurence of cavitation in front of the sonotrode tip, and there are only threee methods suggested for this purpose. It is common to use only one method for measuring the acoustic power, however, in our research we used all threee methods for characterization of our new ultrasonic probe for the neuroendoscopic procedures. Material and Methods: The first method for measuring the acoustic power is based on the electromechanical characterization of the transducer and measuring the diplacement of the sonotrode tip with different amounts of applied electrical power in the active elements of the transucer. The sonotrode tip displacement is measured with optical microscope. The second method is based on measuring the spatial pressure distribution of an ultrasound transmitter produced in an anechoic pool. The acoustic reciprocity principle is used to determinate the derived acoustic power of equivalent sources at frequency components present in the spectrum of radiated ultrasonic waves. The third method is based on measuring the total absorbed acoustic power in the restricted volume of water using the calorimetric method. The temperature increase in the system with known total heat capacity is measured with digital temperature acquisition system at different amounts of applied electrical power and calorimetric system configurations. Results: The results obtained with different methods are not the same as expected. In the electromechanic characterization the electroacoustic efficiency factor is around 40 %, same as one obtained measuring the derived acoustic power in the anechoic pool. The measured output acoustic power using calorimetric method is greater then derived acoustic power, due to large amount of heat energy released in the cavitation process and the electroacoustic efficiency factor obtained with this methods is around 80%. Conclusion:This three methods will enable very good characterization of an ultrasonic surgical device used in medical applications. In the future research, measured parameters will be connected with the amount of produced biological effects in neural and other tissues.


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P2 CLINICAL NEUROSCIENCE

PP06 PREMOTOR AND MOTOR SULCAL COMPLEX AS A POSSIBLE CANDIDATE FOR PREOPERATIVE PLANNING IN PEDIATRIC EPILEPSY SURGERY; EVIDENCE FROM PAST, PRESENCE AND FUTURE IMPLICATIONS Abdel Hadi Fadi1,2, Vasung Lana1, Paladino Josip2, Miklić Pavle2, Radoš Marko1, Kostović Ivica1 1 Croatian Institute for Brain Research, School of Medicine, Universitiy of Zagreb, Croatia, 2Department of Neurosurgery, Clinical Hospital Center Zagreb, School of medicine, Universitiy of Zagreb, Croatia In 1954 Dr. Wilder. G. Penfield described human motor homunculus by stimulating various cortical areas during neurosurgical operations. Until the discovery of MRI in medicine, due to the Paul Lauterbur and Peter Mansfield in 80'‐es, the widely used somatotopic representation of Penfield's motor homunculus has been used in educational, research and surgical planing purposes in a rather abstract and imaginary way. New emerging approaches of MRI in neuroradiology and neuroscience, such as improved postprocessing, DTI and fMRI, have provided hints that parts of the human motor homunculus can be recognized by tertiar sulci. In addittion, significant amount of evidence has emerged from preoperative fMRI and intraoperative motor testing, esspecially planning of motor action, that suggested somatotopic SMA organisation that resembles on Penfield's motor homunculus and can be recognized by demarcation of tertiary sulci of the precentral gyral and sulcal complex. In this anatomical research we have described appearance, variability and MRI usfulness in recognition of menioned tertiary sulci in fetal, pediatric and young adult population based on Croatian Neuroembryological collection. Detail knowledge of sulcal variability and assemblence of terciary premotor and motor sulcy can be of grate importance for pediatric presurgical planning. Frequent epileptic sizuries have adverse affect on developing brain and thus the intractable epilepsy is becoming the more frequent indication for early neurosurgery. As the developing brain retains the capacity for recovery from the resection neurosurgery minimal age limit for surgery becomes obscure. Fallowing and recognizing the external sulcal anatomy, revialed by 3D MRI, can help in presurgical estimation of SMA (in lack of presurgical fMRI and intraoperative stimulation testing) and thus it can contribute to preservation of SMA function in emerging pediatric epilepsy surgery. PP07 GENE POLYMORPHISMS OF ARYLSULFATASE A IN RELAPSE REMITTING MULTIPLE SCLEROSIS: GENOTYPE‐PHENOTYPE CORRELATION AND ESTIMATION OF DISEASE PROGRESSION USING MULTIPLE SCLEROSIS SEVERITY SCORE Koraljka Bačić Baronica1, Kristina Mlinac2, Anton Vladić1, Svjetlana Kalanj Bognar2 1University Department of Neurology, General Hospital Sveti Duh, Zagreb; 2Croatian Institute for Brain Research, School of Medicine, University of Zagreb Introduction: Arylsulfatase A (ASA) is lysosomal enzyme involved in catabolism of cerebroside‐sulfate, major lipid constituent of oligodendrocyte membranes. Various polymorphisms in ASA gene have been described, leading to different levels of enzyme deficiency. Progressive demyelination occurs in metachromatic leukodystrophy (MLD), while the condition of ASA‐pseudodeficiency (ASA‐PD) is suggested to contribute to complex pathogenesis of multiple sclerosis (MS). Genotype‐phenotype correlation is thus useful in estimation of disease severity and progression, and may be analyzed using recently introduced Multiple Sclerosis Severity Score (MSSS) method. Subjects and methods: We have analyzed the frequency of two most common mutations associated with ASA‐PD in 56 patients with diagnosis of relapse‐remitting multiple sclerosis. The presence of mutations was determined by polymerase chain reaction restriction fragment length polymorphism method (PCR‐RFLP). To determine whether there is a difference between disability level and/or disease progression in patients with or without mutations we have estimated disability level using Expanded disability status scale (EDSS) and disease progression using Multiple Sclerosis Severity


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Score for all patients. Correlation of genotypes and disease progression was analyzed by Kruskal‐Wallis test (MSSS test, computer programme, version 3.0). Results: The presence of either one or both mutations was determined in 13 patients (both mutations in 8 patients, only N350S mutation in 4 patients, while 1 patient carried only 1524+95 A‐G mutation). Nine of the mutations carriers had mild disability (EDSS=0‐4.0), 1 had moderate disability (EDSS=4.5‐5.5), and 3 had severe disability (EDSS=6.0‐10.0). Although we found no statistically significant correlation of genotypes and MSSS in the group of 56 MS patients, we have observed that only 3 MS patients who were mutation carriers showed MSSS values lower than 5.000 while in other MS patients‐mutation carriers the MSSS value ranged from 5.267 to 9.453. Conclusion: Since MSSS method takes into account disability as well as the duration of the disease it is more appropriate indicator of disease progression than EDSS which presents only the disability level. MSSS is a powerful method for comparing disease progression in different groups of patients and is proven to be useful for identifying factors that may influence disease progression such as the presence of gene polymorphisms. PP08 QUANTITATIVE EEG IN SCHIZOPHRENIA AND DEPRESSION Begić Dražen1, Jokić‐Begić Nataša2 1Department of Psychiatry, University Hospital Center, School of Medicine, Zagreb, 2Department of Psychology, Faculty of Humanities and Social Sciences, University of Zagreb Introduction: Quantitative EEG (qEEG) has yielded different results in schizophrenia and depression. Research using qEEG has similarly shown altered parameters in both schizophrenia and depression. Since there have been few research attempts comparing patients with schizophrenia and depression using the qEEG parameters, the aim of the present research is to conduct such a comparison. Method: Three groups of subjects were included in study: experimental group I consisting of 30 patients with schizophrenia, experimental group II consisting of 34 patients with depression and a control group with 30 healthy subjects. The diagnoses were established according to the DSM‐IV criteria. EEG was recorded from Fp1, Fp2, F3, F4, F7, F8, T3, T4, P3, P4, O1 and O2 regions. The Fast Fourier Transformation (FFT) method was used to analyze a 10 second period, artifact free. The results are presented in the form of spectral power (|J,V?) for a particular segment of the EEG spectrum (delta, theta, alpha and beta). Results: Results indicate a difference between schizophrenia and depression in the slow rhythms (especially over the frontal regions). Furthermore, in comparison to patients with diagnosed depression, the results indicate a global decrease in alpha activity among patients with diagnosed schizophrenia. Patients were found to differ from healthy subjects in delta, theta and beta rhythms. Conclusions: These results were compared and interpreted in the context of the neuroradiological and neurotransmitter studies of schizophrenia and depression, thus confirming the biological foundation of schizophrenia and depression. Delta and theta rhythms may play a role as a marker in differentiating between schizophrenia and depression. PP09 INCREASE OF BLOOD FLOW VELOCITY IN MEDIAL CEREBRAL ARTERY DURING OBSERVATION OF SUBJECT' S ARM IN A MIRROR Bene Raphael, Lovrenčić‐Huzjan Arijana, Ažman Dražen, Strineka Maja, Budišić Mislav, Vuković Vlasta, Rastovčan Boško, Demarin Vida Department of Neurology KB "Sestre Milosrdnice" Introduction: Mirror illusion consist in the fact that, standing in front of a mirror put in a sagittal plane, with our head on one side and one arm stretched forward, we can see one side of our body reflected as if it were the other side, by mirror visual feedback. The aim of this study was to monitor blood flow changes in medial cerebral artery (MCA) by means of Transcranial Doppler (TCD) in individuals during motor tasks as well as tasks using mirror visual feedback. Subjects And Methods: Eight young healthy volunteers (four male and four female), participated in this study. TCD recording of MCA was done during each task consisting of various motor and visuo‐


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motor activities using mirror illusion. Both MCA mean blow flow velocity (MBFV) were measured while individuals seated in a comfortable chair.The obtained MCA MBFV are presented as baseline values. Results: During the illusion of motoric hand activation, when the subject is making right hand flexions and watching it's reflexion in the mirror ,while the left hand is immobile, increase of mean blood flow velocity of contralateral MCA was observed (task 3 +4,5% than in baseline values, p=0,017). Furthermore, when the subject made left hand flexions while watching the reflection of the immobile right hand in the mirror, there was increase of MBFV in right MCA (+5.6% than in baseline values p=0,044), more pronounced than during the illusion of motoric hand activation (task3) and less than during direct vision of hand flexion (task 2 +6.3%than in baseline values p=0,005). Conclusion: Our data showed that visual illusion of action, as well as direct action observation can increase mean blood flow value in MCA, which brings forward the possible usage of mirror illusion as a tool for motoric neurorehabilitation. PP10 THE MOST COMMON SYMPTOMS OF PINEAL GLAND CYST Bošnjak Jelena, Miškov Snježana, Hećimović Hrvoje, Čović Negovetić Ružica, Demarin Vida Department of Neurology, University Hospital "Sestre milosrdnice", Zagreb INTRODUCTION: Since the introduction of magnetic resonance imaging (MRI), between 1.26% and 4.3% of patients examined for various neurological reasons had pineal gland cyst. The most common symptoms result from compressive effect of the surrounding structures. PATIENTS: We present 46 patients examined from February 2007 to January 2009 who had cystic lesion of the pineal gland detected on MRI of the brain. They had no concomitant brain damage or other underline diseases. RESULTS: Patients examined were 7‐65 years, mostly complained for headache (21), epilepsy (15), visual disturbances (5) and vertigo (8) as isolated symptoms or in clusters. The size o pineal gland cyst varied from 4x5x5 mm to 15x16x21mm. In 7 patients compression on the superior collicula was described on MRI. Patients presenting with epilepsy had: tonic clonic seizures (4), partial secondary generalized (4), simple (2) and one complex absance, complex partial seizures (3) and generalized tonic seizures in one patients. All children underwent evaluation of neurosurgeon, 4 of them were operated becaming symptom free. All patient with epilepsy were put on antiepileptic drugs (AED) (mono or politherapy) and became seizure free. Patients who were not surgicaly treated all underwent follow up MRI and had no cyst enlargement. CONCLUSION: In our selected population 46 patients had cystic lesion of the pineal gland on MRI with headache, epilepsy, vertigo and visual disturbances as presenting symptoms. Four patients were operated and became symptom free. Patients with epilepsy were stabilized on AED. PP11 AGE‐DEVELOPMENTAL STAGE AND SEVERITY OF TRAUMA RELATED SYMPTOMS, ANXIETY AND DEPRESSIVE SYMPTOMS IN PARTICIPANTS WHO LOST THEIR FATHERS DURING THE WAR IN CROATIA Dijanić Plašć Ivana1, Poljarević Sanja2, Lončar Mladen3, Henigsberg Neven3

1Dom za djecu i odrasle žrtve obiteljskog nasilja „Duga ‐ Zagreb", Zagreb,2Privatna neuropsihijatrijska ordinacija, Pula, 3Hrvatski institut za istraživanje mozga, Zagreb A number of authors have noted that older children are more vulnerable than younger children to the psychological effects of trauma. The younger child's psychological response resonates with parental response as they have less cognitive capacity to independently evaluate dangers. Some researches explained those with circumstances that surround the death can prevent children and adolescents from grieving the loss, thus increasing the risk of long‐term consequences. There are many explanations for such findings such as social, community and family cohesiveness and support systems, but it could be also temperamental and biological factors that modulate stress response. Children of different ages will experience a traumatic event in a different ways. The most important in generalization the research findings is recognizing that children of different ages think differently,


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act differently and have different emotional functioning. Experiences that are extremely traumatic to an adult may be perceived by a young child as something that is not so frightening. On the other hand, the fear that the child feels will more frequently be a reflection of that of the adult rather than generated by the child's own perception of the event. So, the individual experience of the trauma is age dependent. Our study focused on children who lost their fathers in conditions of war. The participants were at the time of interviewing mostly adolescents or young adult which means that some of them were not even born when their father died, some of them were very young children, preschoolars and some of them were adolescents. Our aim was to explore the association between age‐developmental stage and severity of trauma related symptoms, anxiety and depressive symptoms in participants who lost their fathers during the war. The study included 114 people who lost their fathers during the war in Croatia, who came to the physical and psychiatric examination organized by Ministry of family, war veterans and intergenerational solidarity. Data were collected using a structured clinical interview which also included socio‐demographic data. Data about former and current psychiatric symptoms were collected using the following instruments: Clinician Administrated PTSD Scale, Hamilton anxiety scale, Hamilton depression scale. Results showed that there was significant correlation between age and results on used scales. The participants who lost their fathers in very young age or even before they were born showed less trauma symptoms, less anxiety and depressive symptoms then participants who lost their fathers in older age. The study confirmed that the individual experience of the trauma of losing the father in war circumstances is age dependent. PP12 AUTISM IN CHILDHOOD‐EARLY SCHIZOPHRENIA OR SOMETHING ELSE? ATYPICAL ANTIPSYCHOTIC AS A DRUG CHOICE Dodig‐Čurković Katarina1, Čurković Mario2, Dunja Degmečić, Dodig‐Radić Josipa3, Radić Mislav3 1University departement for child and adolescent psychiatry,University departement of psychiatry Osijek; 2Family medicine departement, Health center Osijek; 3University departement of internal medicine, University health center Split INTRODUCTION: Autism is a pervasive developmental disorder characterised by impairment in social interaction and communication, with unusaul behaviour. Genetic factors are predominant in autism patogenesis1. The causes of autism spectrum disorders are unknown, although genetic and enviromental influence have been implicated.2 There is increasing evidence that people with autism spectrum disorder have abnormalities in the serotonergic system.3 Also, still there are limitted options for pharmacological therapeutic interventions in chlidren with autism disorders, some studies showed that risperidone as a atypical antypsichotic, may be effective for the treatment of people with autism and intellectual disabillities.4 The most studied antypshochotic drugs include haloperidol and risperidone.5 Haloperidol and risperidone are the most widely studied drugs for reducing symptoms in children and adolescents with autism. In low dosages, they have been shown to reduce repetetive behaviours /stereotypies/ and social withdrawal, as a well, as a number of related symptoms, such as a hyperactivity, agression, self‐abuse behaviour, liability of mood and irritability.6 All the listed symptoms have appeared in adult patients with schizophrenia, where also with the application of atypic antipsychotics may affect on desribed clinical picture. MATERIALS AND METHODS: We displayed two cases of autism in boy and girl aged 14 and 16 years, with confirmed diagnosis of autism according to DSM‐IV i ICD‐10 classification criteria. Girl was completely working and social of malfunction behaviour, with often agressive excesses, stereotypes, she was proned to self‐destructive forms of behaviour, mutistic, self‐injured herself, etc. Unlike her, the boy was described as a "type of well functioning autism", he have attended a school on an adjusted programme, he was hyperactive, with behaviour problems and learning difficulties. Both children were under treatment with risperidone during few years, girl with dose of 4 mg per day, and boy with dose of 2 mg per day. RESULTS: with the application of risperidone in girl, we have achieved reduction of psychomotoric symptoms and reduction of hetero‐agressive and self‐destructive behaviour, but largely were still


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present stereotypes, social isolation, mutistic behaviour, similarly to negative phase of schizophrenia in adults. In boy we have also achieved reduction of psychomotoric symptoms, with improvement in contact with his surrounding, he had less learning problems and he has felt familiar not only with his mother but with other persons. CONCLUSION: autism in childhood would be according to clinical picture and the manner of response to treatment with atypic antipsychotics may represent a form of schizophrenia in juvenile age or could be an introduction in further development of psychotic process in adult age. PP13 SEASONALITY OF SCHIZOPHRENIA BIRTH BY SCHIZOPHRENIA TYPES Henigsberg N1,2, Kalember P2,3, Folnegović Šmalc V1, Hrabač P4, Lončar M5

1Psychiatric Hospital Vrapče, University Department of General and Forensic Psychiatry and Clinical Psychophysiology, Zagreb, Croatia; 2Department of Neuropharmacology and Behavioural Pharmacology, Croatian Institute for Brain Research, Medical School, University of Zagreb, Zagreb, Croatia; 3Polyclinic Neuron; 4Croatian Institute for Brain Research, Medical School, University of Zagreb, Croatia; 5Zagreb University Hospital The definite etiology of schizophrenia is still unknown, although there are many theories trying to explain it. It is considered that seasonality of schizophrenia could clarify the etiology of this illness. There are very few studies in differences in the birth seasonality regarding the type of schizophrenia but the results of these studies are inconsistent. Our research is conducted in the group of patients with the diagnosis of schizophrenia, who were born in the period between January, 1st 1950 and December, 31st 1960 and who were hospitalized in medical institutions in republic of Croatia by the year 1990. Data from Register of psychosis of the Croatian National Institute of Public Health and Central Bureau of Statistics were used. There were 3.257 subjects included in this study. Birth rate of schizophrenia patients with regards to the month of birth does not significantly deviate in paranoid, catatonic, hebephrenic, residual, simple, undifferentiated and other schizophrenia. In conclusion, we did not find any significant seasonality trends with regards to the different types of schizophrenia. However, we plan to expand the present study to younger persons and to persons hospitalized after 1990. Analysis of this new data will clarify present findings and perhaps shed some new light on this topic. PP14 1‐H MRS CHANGES IN DORSOLATERAL PREFRONTAL CORTEX AFTER DONEPEZIL TREATMENT IN PATIENTS WITH MILD TO MODERATE ALZHEIMER'S DISEASE Henigsberg N1,2, Kalember P2,3, Hrabač P4, Radoš M4,5, Bajs M5, Radoš M4,5, Kovačić Z4, Lončar M5, Madžar T4 1Psychiatric Hospital Vrapče, University Department of General and Forensic Psychiatry and Clinical Psychophysiology, Zagreb, Croatia; 2Department of Neuropharmacology and Behavioural Pharmacology, Croatian Institute for Brain Research, Medical School, University of Zagreb, Zagreb, Croatia; 3Polyclinic Neuron; 4Croatian Institute for Brain Research, Medical School, University of Zagreb, Croatia; 5Zagreb University Hospital Magnetic resonance spectroscopy (MRS) noninvasively provides information on the concentration of some cerebral metabolites in vivo. Among those measurable by proton magnetic resonance spectroscopy (1H‐MRS), N‐acetyl‐aspartate (NAA) is decreased, and myo‐inositol (mI) and choline (Cho) levels are increased in patients with Alzheimer's disease (AD). Donepezil, an acetylcholinesteraze inhibitor, has proven effect on cognitive symptoms in patients with AD. In previous studies, treatment response was associated with an increase of NAA and NAA/Cr in the parietal lobe and hippocampi. Correlation of longitudinal changes of 1H‐MRS detectable metabolites in dorsolateral prefrontal cortex (DLPFC) with clinically observable changes were not researched. The objective of this non‐interventional study is to assess whether changes in 1H‐MRS measurable metabolites correlate with clinical outcome after donepezil treatment.


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Twelfe patients with mild to moderate AD were evaluated during 26 weeks of donepezil treatment. 1H‐MRS parameters were assessed before and after 26 weeks of donepezil treatment. Cognition was assessed with Alzheimer's Disease Assessment Scale cognitive subscale. A significant increase in NAA/Cr ratio and significantly lower decrease in mI/Cr ratio were found in AD patients with positive treatment response. The results of this study indicate possible modest donepezil effect on prevention of neuronal functional deterioration in DLPFC which correlates with clinical outcome and point the use of 1H‐MRS as technique of help in assessment of drug effect. PP15 COULD A NEUROLOGICAL DISEASE BE A PART OF MOZART'S PATHOGRAPHY? Ivkić Goran1, Erdeljić Viktorija2 1Croatian Institute for Brain Research, University of Zagreb, Šalata 12 10000 Zagreb, 2University Hospital Zagreb, Division of Clinical Pharmacology, Department of Medicine, Kišpatićeva 12 10000 Zagreb, Croatia As expected, since we recently celebrated the 250th anniversary of the birth of Wolfgang Amadeus Mozart, there has been again a renewal of interest in his short but intensive life, as well as in the true reason of his untimely dead. Mozart lived and died in time when the medical knowledge was based mostly on subjective observations, without the basics of standardized medical terminology and methodology. This leaves a great space for hypothesizing about his health problems, as well as about the cause of his death. The medical academic community attributed to Mozart approximately 150 different medical diagnoses. There is much speculation on the cause of Mozart's death: uremia, infection, rheumatic fever, trichinelosis, etc. Recently some authors have raised the question about a possible concomitant neurological disease. According to available records, Mozart has shown some elements of cyclotimic disorder, epilepsy and Gilles de la Tourette syndrome. Furthermore, the finding of a temporal fracture on (allegedly) Mozart's skull, gives a way to speculations about the possibility of a chronic subdural hemathoma and its compressive effect on the temporal lobe. Despite numerous theories on Mozart's patography that also include a concomitant neurological disorder, the medical and history records about Mozart's health status indicate that he probably had suffered from an infective illness, followed most likely by the reactivation of rheumatic fever, which was followed by strong immunologic reaction on the last days of his life. Taking all the above into consideration, it is reasonably to conclude that Mozart's neurological disturbances were caused by the intensity of the infective disease, and not primarily by a neurological disease. PP16 CORRELATION OF VERBAL MEMORY AND ILLNESS INSIGHT IN THE SCHIZOPHRENIC PATIENTS Janović Š1, Bajs M1, Đorđević V1, Hrabač P2, Radonić E2, Heningsberg N2

1University department for psychiatry, University Hospital Center Zagreb, Zagreb Medical School, Zagreb, Croatia, 2Croatian Institute for Brain Research, Medical School, University of Zagreb, Croatia Disturbances of the illness insight in the schizophrenia is one of the core problems when addressing treatment and outcome of schizophrenia. Aim of the study was to investigate cognitive functions like verbal memory in schizophrenic patients and its correlation to the illness insight. Participants (N= 60) underwent neurocognitive testing, clinical scales for psychotic symptoms and behavioral scales. Results: Patients with poorer insight in to illness had worse results in neurocognitive testing, especially verbal memory. They also had earlier onset of illness, higher number of psychiatric hospitalizations and involuntary psychiatric hospitalizations, and less regular psychiatric treatment. They were also more often unemployed, unmarried and without children. There were no differences in BPRS and suicidal behavior among groups. Conclusion: Impairments in cognitive functioning could influence treatment decisions in schizophrenic patients, leading to poorer compliance and coping with illness and resulting in less favorable treatment options. As cognitive functioning influences variety of quality of life domains, including insight and compliance, cognitive deficit is important target for therapeutic intervention in schizophrenic patients.


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PP17 AUTOMATIC SPINDLE DETECTOR FOR INFANT DATA Jerončić Ana1, Achermann Peter2 and Đogaš Zoran1

1Department of Neuroscience, University of Split School of Medicine, Split, Croatia, 2Institute of Pharmacology and Toxicology, University of Zurich, Zurich, Switzerland Introduction: Sleep spindles are transient EEG events which have been used as a tool in automatic sleep stage scoring. In addition, spindles have been used as potential markers of sleep‐related memory consolidation and different neuropathological conditions. Here we report the development of the robust and precise automatic spindle detector. Methods: Algorithm Trnoružica (Sleeping Beauty) was developed on the training dataset of visually detected spindles from all‐night EEG recordings of 2 and 9 month old infants (N=10). One hour of continuous EEG recordings, C3 A2‐derivation, was used in each case. Standard signal processing methods including filtering and spectral analysis (FFT) were used in line with decision‐tree modeling technique for the algorithm development. Custom made scripts were written in MATLAB 7.4 software for this purpose. Results: Descriptor values used in the detection process were successfully calculated from the statistical properties of the EEG recordings tested. Therefore, Trnoruzica was applied to different EEG derivations of infant, as well as adult data and was found to be robust to the source of data. When rigorously tested on continuous EEG recordings, with either very low or high frequency of visually detected spindles, detector has shown on average 89% of sensitivity and 99% of specificity, with positive prediction value of 60%. Conclusion: Although primarily developed for the infant data, a wider spectra of application and the precision makes the Trnoružica algorithm a promising general tool for spindle detection. PP18 IS THERE AN IMPACT OF GENETIC MARKERS IN AMYOTROPHIC LATERAL SCLEROSIS PATIENTS ON COGNITIVE IMPAIRMENT? Liščić RM1, Muck‐Šeler D2, Mustapić M2, Babić A2, Stukovnik V3, Zidar J3 1Institute for Medical Research and Occupational Health, Zagreb, Croatia, 2Division of Molecular Medicine, Rudjer Bošković Institute, Zagreb, Croatia, 3Institute of Clinical Neurophysiology, University Medical Centre Ljubljana, Slovenia Background: The overlap between cognitive impairment and behavioral features in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia is demonstrated in up to 50% of ALS patients. Behavioral features are mostly due to changes in serotonergic and catecholaminergic system. The aim of the study was to compare gene polymorphisms coding for proteins involved in serotonin and catecholamine metabolism and function with the emphasis on executive function in ALS patients. Materials and methods: The prospective study included 16 ALS patients (10 male, 60.5±5.8 years) defined by El Escorial Criteria. Genetic markers: ‐1021 C/T polymorphism of DBH gene, 102 C/T polymorphism of 5‐HT2A receptor gene, vall58met polymorphism of COMT gene and val66met polymorphism of BDNF gene were correlated with two tests of executive functions, Controlled oral word association and Tower of London (TOL). Results: The frequency of genotypes for particular gene polymorphism were: COMT (GG‐33%, GA‐53%, AA‐14%), BDNF (GG‐73%, GA‐20%, AA‐7%), DBH (CC‐47%, CT‐47%, TT‐6%) and 5‐HT2A (CC‐30%, CT‐60%, TT‐10%). 57% of patients showed deficient word generation capability. 21% of patients were impaired on TOL Total move score and 33% of patients on TOL Total rules violation score. No significant relationship was found between genes polymorphism and variables of executive functional tests. Conclusion: The preliminary results showed no correlations between gene polymorphisms and variables of executive functional tests. A sizable proportion of ALS patients' showed behavioral and cognitive changes within a spectrum of frontotemporal impairment. Further studies on a larger sample, however, are needed in order to confirm it.


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PP19 LONG‐TERM MORTALITY IN PERSONS IMPRISONED DURING THE WAR IN CROATIA Lončar M1, Henigsberg N2,3, Hrabač P4, Gregurek R5, Grujić I1, Erceg M6, Čorić T6 1Ministry of the Family, Veterans' Affairs and Intergenerational Solidarity, 2Psychiatric Hospital Vrapče, University Department of General and Forensic Psychiatry and Clinical Psychophysiology, Zagreb, Croatia, 3Department of Neuropharmacology and Behavioural Pharmacology. Croatian Institute for Brain Research A long‐time mortality of persons imprisoned in Serbian camps and prisons during the war in Croatia was observed. We gathered data about 6.237 prisoners. The prisoners were followed from their release from the camps to December 2006. Among the persons studied, men were more numerous than women (80,86% to 19,14%). Also, there were more soldiers (60,11%), than civilians (39,89%). Prisoners were released from camps or prisons at the average age of 37,8 years, after they spent a mean period of 103 days in captivity. Over the above mentioned time period, 467 ex‐prisoners died. Average age at the time of death was 61,9 years. The most common causes of death were ischemic heart disease, injuries, cerebrovascular disease, neoplasms and liver diseases. We found an inverse correlation between number of days spent in prison and the age of death. We conclude that population of ex‐prisoners differs significantly from general population in terms of age and causes of death. PP20 MINIMALLY INVASIVE ANEURYSM TREATMENT AT SCHOOL OF MEDICINE UNIVERSITY OF ZAGREB Mrak G, Paladino J, Radoš M, Jednačak H Department of neurosurgery, School of Medicine, University of Zagreb Aim of the study: The results of neurosurgical treatment of patients with cerebral aneurysms admitted at University Hospital Center Zagreb are presented in this review. Even with introduction of coilling in aneurysm treatment there is still place for neurosurgical approach. Methods: In the period from May 1996 to April 2008, keyhole approach was used in surgical treatment of 1003 patients with various intracranial pathology. During the same time 811 patients with 947 aneurysms were operated on, and further 160 patients were treated endovascularly. The majority of 811 patients with aneurysm were operated on through small keyhole craniotomy in different locations (652). Most of them were operated on through eyebrow‐keyhole craniotomy. The remaining 161 patients were operated on using larger craniotomy. Results: Good outcome was achieved in most of the microsurgically treated patients. Excellent or very good outcome was obtained in 84% patients from keyhole craniotomy group, and 79% of patients in standard craniotomy group. The mortality rate in keyhole group was 0,61%, and in standard craniotomy group 1,87%. Conclusions: Since introduction of endovascular techniques in aneurysm treatment, conjoined cerebrovascular team decide upon treatment options individually for each patient harbouring intracranial aneurysm. Keyhole approach with aid of lumbar drainage, endoscopic assistance and evoked potential monitoring is safe and reliable treatment option for intracranial aneurysms. According to our results, microsurgery still has an important role in aneurysm treatment. PP21 HIPPOCAMPAL VOLUME IN SCHIZOAFFECTIVE DISORDER Radonić E, Henigsberg N, Radoš M, Kalember P, Bajs‐Janović M, Folnegović‐Šmalc V Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Šalata 12, 10000 Zagreb, Croatia Introduction: In studies of the neurobiological basis of psychotic disorders, schizoaffective disorder is sometimes included as part of schizophrenia patient group and sometimes as part of bipolar disorder patient group. Since most studies report hippocampal volume reduction in schizophrenia and not in bipolar disorder when analyzed without the inclusion of schizoaffective disorder patients, the aim of the study was to analyze hippocampal volume in schizoaffective disorder as a separate group.


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Materials and methods: Hippocampal lobe volumes were estimated using high resolution magnetic resonance imaging in 60 subjects, 15 subjects in each patient and in one healthy subjects' group. Results: Hippocampal volume in both hemispheres is statistically significantly reduced in schizophrenia and schizoaffective disorder compared to healthy control group. There is no statistically significant difference in hippocampal volume between bipolar disorder and healthy control group. Conclusion: Further studies are needed to compare other frequent neurobiological findings in schizophrenia, schizoaffective and bipolar disorder. Even though these psychotic disorders share many clinical features, their clinical differences may be substantiated by differences in structural neurobiological findings. In further studies these disorders should be analyzed separately. Key words: hippocampal volume, magnetic resonance imaging, shizoaffective disorder PP22 NEUROENDOSKOPSKO LIJEČENJE HIDROCEFALUSA Rotim Krešimir, Božić Boris, Kudelić Nenad, Kčira‐Fideršek Vječeslav, Saftić Robert Klinika za neurokirurgiju "Sestre milosrdnice", Zagreb, Hrvatska Operacije ekstrakranijske derivacije cerebrospinalnog likvora najučestalije su primjenjivani zahvat u liječenju hidrocefalusa. Ti su neurokirurški zahvati praćeni brojnim mehaničkim i biološkim komplikacijama ugradnje stranog materijala, a postignuti rezultati u liječenju hidrocefaličnih bolesnika nisu posve zadovoljavajući. Usavršavanje endoskopskih zahvata i minijaturizacija endoskopske opreme zadnjih godina aktualizira metodu endoskopske ventrikulostomije. Endoskopska ventrikulostomija treće mozgovne klijetke postupak je izbora u liječenju opstrukcijskog hidrocefalusa. Tim se neurokirurškim zahvatom izbjegava ugradnja silikonskog derivacijskog uređaja u bolesnikovo tijelo, čime se značajno prevenira mogućnost biologijskih i mehaničkih komplikacija. Endoskopskom ventrikuilostomijom treće mozgovne klijetke istodobno se uspostavlja gotovo fiziologijsko protjecanje likvora iz nutarnjih u izvanjske subarahnoidalne prostore lubanjske osnovice, za razliku od derivacijskih zahvata kojima se palijativno i najčešče samo privremeno rješavaju simptomi hidrocefaličnih bolesnika. Najučestaliji razlog za primjenu neuroendoskopije je supratentorijski triventrikularni nekomunicirajući hidrocefalus, najčešće prouzročen stenozom ili okluzijom mezencefaličnog akvedukta. Prezentirano je neuroendoskopsko liječenje opstrukcijskog hidrocefalusa Klinike za neurokirurgiju Kliničke bolnice "Sestre milosrdnice" u Zagrebu. PP23 FUNCTIONAL DIAGNOSTICS OF READING DIFFICULTIES IN DYSPHASIC ADULTS Runjić Nađa, Vranić Đurđica SUVAG, Zagreb Dysphasia is one of speech disordes in which there is impairment of the power of expression by spoken language, writing, or signs, or impairment of the power of comprehension of spoken or written language. Functional diagnostics of such patients is dirrected to the positive rehabilitation outcome. The objective of this study was to present the minimal diagnostic program for reading disabilities in patients with sensomotor dysphasia. Ten patients aged 40‐80 were tested. Control group consisted of 10 healthy persons matched by age, gender and nonverbal status. Complete diagnostic evaluation was performed included ophtalmological, otoneurological, evoked auditory and visual potentials, logopedic, psychological and psychiatric evaluation. The results show the positive correlation betweeen: 1. auditory synthesis and analysis results and auditory brainstem potentials findings, 2. visuomotor function results and visual evoked potentials. Minimal functional diagnostic program for dysphasic patients with reading difficulties must consist of neurological, logopedic and psychological testing. According to the results of psiholinguistic abilities evoked potentials testing ( auditory and/or visual one) will be done for the rehabilitation purposes.


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PP24 CURRENT APPROACHES TO THE PHARMACOTHERAPY OF OPIATE ADDICTION Sakoman S1 and Jernej B2

1Department of Psychiatry, University Hospital "Sestre milosrdnice", Vinogradska 29, HR‐10000 Zagreb, CROATIA; 2Laboratory of Neurochemistry an Molecular Neurobiology, Rudjer Bošković Institute, Bijenička 54, HR‐10000 Zagreb, CROATIA Partial agonists of neurotransmitter receptors, often called "stabilizers", act somewhere between full receptor agonists and silent receptor antagonist, theoretically boosting defficient but also inhibiting excessive neurotransmitter activity. Pharmacotherapy of opiate dependence, besides traditional substitution with methadone ‐ the opioid mu‐receptor full agonist (and possible adjuvant detoxification with opioid antagonist naltrexone) is currently oriented toward partial agonists of opioid receptors. Buprenorphine (Subutex) demonstrates partial mu‐receptor agonism and seems a good alternative to methadone substitition in most cases of opiate addiction. Newer derivative of buprenorphine ‐ Suboxone besides buprenorphine contains about 25% of opiate antagonist naloxone which prevents intravenous abuse. Results of recent clinical experience with administration of Subutex in opiate addiction will be presented and indications as well as contraindications for its use will be discussed. PP25 ANTHROPOMETRIC VARIABLES AND MEAN OXYGEN SATURATION AS PREDICTOR FOR CLINICAL CONSEQUENCES IN OBSTRUCTIVE SLEEP APNEA PATIENTS Ševo Vana1, Selimović Mirnes1, Vrkić Dina1, Račić Goran2 and Đogaš Zoran1

1Sleep Lab for Clinical Neuroscience, Department of Neuroscience, University of Split School of Medicine, 2ENT Department, University Hospital Split, Split, Croatia Introduction: Obstructive sleep apnea (OSA) is a common medical condition that occurs in approximately 5% to 1 5% of the population. Diagnosis of OSA is based on polysomnography, and its severity is measured with Apnea‐Hypopnea Index (AHI). OSA is characterized by recurrent, complete or partial airway closure during sleep. It is potentially hazardous condition and appropriate treatment should be strongly advised to all OSA patients. Untreated OSA leads to excessive daytime sleepiness, diminished performance and overall poor quality of life. The role of OSA in promoting atherosclerosis, hypertension, inflammation and a procoagulant state has now been established. Aim of this study was to explore the correlations of anthropometric variables and sleep parameters with clinical symptoms such as excessive daytime sleepiness in the patients with obstructive sleep apnea who underwent the polygraphy recordings in the Sleep Laboratory in Split. Materials and methods: The data were collected from 120 patients tested between the years 2005 and 2007. AHI, DesI and mean oxygen saturation as well as other standard sleep parameters were determined with the use of the same diagnostic device PolyMESAM (MAP, Germany). To determine clinical symptoms, extensive questionnaires including the standard Stanford Sleepiness Scale, and Epworth Sleepiness Scale were used. Results: Mean oxygen saturation significantly correlated with ESS score (r=‐0,22, P=0,036). Both BMI and Neck circumference exhibited strong association with AHI and DesI. Heart Rate Variation Index was significantly positively associated with AHI (r=0,28, P=0,02). There was a strong correlation between the AHI and DesI in our patients. Epworth Sleepiness Scale score did not exhibit significant association with PSG diagnostic criteria AHI and DesI. Conclusion: Mean Sa02 significantly negatively correlated with the Epworth Sleepiness Scale Score. Anthropometric features like BMI and neck circumference both strongly correlated with AHI and DesI, and are very useful in selecting patients for OSA diagnostic procedures. Extensive questionnaires (Epworth Sleepiness Scale and Stanford Sleepiness Scale) did not seem to be useful diagnostic measures in our patients. Due to importance of DesI and Mean oxygen saturation, pulse oxymetry is very useful as a screening method for diagnosis of OSA.


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PP26 THE CHARACTERISTICS OF VISUAL EVOKED POTENTIALS IN SPEECH IMPAIRED CHILDREN Vranić Đurđica, Runjić Nađa SUVAG, Ul. kneza Ljudevita Posavskog 10, Zagreb Previous studies have resulted that some speech impaired children show slower maturation of central visual function. Evoked visual potentials testing with cortical cartography is an useful diagnostic method for the visualisation of functional changes in visual pathway. The aim of this study was to investigate the characteristics of visual evoked potentials in children with delayed speech development. Twenty speech impaired preschool children aged 6‐7 were tested, divided to the verbal results ( Reynell Developmental Language Scale). Control group consisted of 10 healthy children, matched by age, gender and nonverbal status. Complete diagnostic evaluation were performed included ophtalmological, otoneurological, logopedic and psychological evaluation. Subjects and controls were examined by checkerboard pattern reversal visual evoked potentials (VEP) according to the 2004. European standards. Cortical cartography was simultaneously performed by Neuroscan 32‐electrode system using Scan 4.3 software for data analysis. The results show positive correlation among N 135 wave characteristics (thalamocortical level) in visual evoked potentials and psycholinguistic abilities (Psycholinguistic language Acquisition). Children with immature visuomotor function show signifficantly shortened amplitude and delayed latency of N 135 wave during monocular and binocular stimulation. Speech impaired preschool children with immature visuomotor function should be evaluated by visual evoked potentials with the purpose of efficient rehabilitation work. PP27 EEG POLYGRAFIC STUDY OF MATURATIONAL DIFFERENCES BETWEEN TWINS Vučinović Mirjana, Ursić Anita, Rešić Biserka Clinical Hospital Centre Split, Pediatric Clinic, Children Sleep Laboratory, University of Split, School of Medicine Objective: Multiple gestations are stressed during the third trimester because of intrauterine crowding and limited uteroplacental supply. They increase the likelihood of preterm birth but its influence on rate of maturation or complications of prematurity has been controversial. The purpose of this pilot study is to implement a recently completed analysis of a prospective study to asses the effect that multiple pregnancies exert on the maturation of the developing brain. Methods: Data from prospectively recorded 10 twin sets, born prematurely ( mean 36 gestational week ; range 3 1‐38 GW) in the University Hospital Split during period from January 2009 to Jun 2009 were analysed. They were selected according to the following criteria: normal pregnancy and birth: 1 min Apgar: 7 or above; 5min Apgar, 9 or 10; negative post‐natal physical and neurological examinations. We evaluated EEG polygrafic records obtained on the mean = 38 week ; range 35‐41 week postmenstrual age( PMA), and on mean = 43 week ; range 41‐46 week PMA . We compared duration of the sleep cycles and the sleep epochs ; active sleep (AS), quite sleep ( QS ), indeterminate sleep (IS ) among twins, using the Man Whitney test. Results: Forty EEG polygrafic records were analysed. At the first measurement lenght of IS was 2,4 fold higher in the first twin (mean =13min ; range 5‐16 min) comparing with second twin (mean = 5 min ; range 2‐14 min ) ( p = 0, 005 ). Percent ‐ time IS sleep was significantly higher in the first twin ( mean = 16,9 %; range 5,4 ‐ 22 % ) than in the second one ( mean 6,5 % ; range 2,2 ‐ 28 % ). The IS sleep proportion in the entire sleep cycle was 2,6 fold higher in the first twin ( p= 0, 028). Percent ‐ time QS was 2,5 fold longer in the second twin than in the first one ( 33,2 vs. 13,4 % ; p = 0, 054). Duration of sleep epochs among twins at the second measurement were similar. Conclusion: Present data (longer IS and shorter QS in the first twin at the first recordings), obtained on the small number of the twins, showed that firstborn twin showed more immature EEG poligrafie pattern compared with second twin in the first recording, but not at the second recording obtained four weeks later. It might be suggested that the maturational differences among twins exist in utero and shortly after birth, they disappear until the end of the first month of the postnatal life.


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PP28 HYDROXYAPATITE CERAMICS IN MULTILEVEL CERVICAL INTERBODY FUSION‐IS THERE A ROLE? Vukić M1, Walters CB2, Radić A3, Mihaljević D1, Jednačak H1, Marasanov S1, Jurjević I4 1Department of Neurosurgery, School of Medicine University of Zagreb, 2Department of Neurosurgery, New York University, New York, USA, 3Department of Radiology, General Hospital Karlovac, 4Department of Pharmacology, School of Medicine University of Zagreb Introduction: This study is performed to evaluate efficacy of hydroxyapatite grafts in multilevel cervical interbody fusion at one year follow‐up. Methods: Eighty‐six patients with degenerative cervical disc disease underwent 224 cervical interbody fusion procedures in which either Smith‐Robinson or Cloward type hydroxyapatite grafts were used. Indications for surgery included radiculopathy in 38 cases, myelopathy in 20 cases and myelo‐radicular combination in 28 patients. In 65 out of 86 patients fusion was followed by anterior instrumentation (plating). Postoperatively patients were followed for a mean of 15.64 (range 11‐23.3) months. All patients underwent radiography to evaluate fusion and the axis curvature. Results: Excellent clinical results (86%), described as complete or partial relief of symptoms with full return to activity, were obtained in patients with radiculopathy. There were 5 grafts mobilization and one graft fracture. Two grafts extruded in non‐instrumented patients and required repeat surgery. There were other three reoperations due to the hardware problem and consequently pseudoarthrosis. One year fusion rate was obtained at 86 % for two‐level surgery, 80.1 % for three‐level surgery and 74 % for four‐level surgery. Mean (SD) hospital stay was 3.8 (0.7) days. Conclusion: Hydroxyapatite grafts can be very effective synthetic material for multilevel cervical interbody fusion. It is characterized by a high fusion rate and a small percentage of graft‐related complications especially if fusion procedure is followed by plating.

P3 NEUROPHARMACOLOGY PP29 CHANGES OF BRAIN METABOLITES MEASURED WITH MAGNETIC SPECTROSCOPY IN ANTIDEPRESSANT RESPONDERS WITH COMORBID MAJOR DEPRESSION AND POSTTRAUMATIC STRESS DISORDER Henigsberg N1,2, Bajs M1, Hrabač P3, Kalember P1,2, Radoš M4, Radoš M3, Radonić E3 1Psychiatric Hospital Vrapče, University Department of General and Forensic Psychiatry and Clinical Psychophysiology, Zagreb, Croatia, 2Department of Neuropharmacology and Behavioural Pharmacology, Croatian Institute for Brain Research, Medical School, University of Zagreb, Zagreb, Croatia, 3Croatian Institute for Brain Research, Medical School, University of Zagreb, Croatia, 4Clinical Institute of Diagnostic and Interventional Radiology, University Hospital Center Zagreb, Zagreb Medical School, Zagreb, Croatia

Introduction: In a present pilot study, performed on 11 subjects, we studied proton magnetic resonance spectroscopy (1H‐MRS) changes in early to intermediate (3‐6 weeks) responders to antidepressant treatment with selective serotonin reuptake inhibitors (SSRIs). All subjects had diagnosis of major recurrent depression comorbid to posttraumatic stress disorder (PTSD). Subjects and methods: Magnetic spectroscopy was done in the region of dorsolateral prefrontal cortex on a 3T MRI‐unit. Participants were selected out of the larger sample due to an early response to antidepressant treatment within 3‐6 weeks, measured with Beck Depression Inventory (BDI). We measured levels of neuronal marker N‐acetyl‐aspartate (NAA), choline (CHO) and creatine (CRE). Results: There was no difference in NAA/CRE ratios at the first and second spectroscopy (p=0,514). However, CHO/CRE ratios showed different trend with mean value at the first spectroscopy of 1,07 (sd=0,12) while mean value at second was 1,22 (sd=0,08) with statistical significance with p‐value of 0,003. In conclusion, significant increase in choline to creatine ratio from the first to the second spectroscopy during the antidepressant treatment, compared to almost identical values of NAA to creatine ratio, suggests increased turnover of cell membranes as a mechanism of the early response to the antidepressant drug therapy.


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PP30 PROLONGED EXPOSURE OF RECOMBINANT GABA‐A RECEPTORS TO DIAZEPAM INDUCES DIFFERENTIAL EFFECTS ON FUNCTIONAL COUPLING AND EXPRESSION OF BENZODIAZEPINE BINDING SITES Švob Štrac D, Jazvinšćak Jembrek M, Vlainić J, Peričić D Ruđer Bošković Institute,Laboratory for Molecular Neuropharmacology,Division of Molecular Medicine Bijenička cesta 54, 10 000 Zagreb, Croatia Variety of drugs, including clinically relevant benzodiazepines, exerts most of their pharmacological effects via GABA‐A receptors, the major fast inhibitory neurotransmitter receptors in the mammalian brain. The aim of this study was to further explore the mechanisms that underlie adaptive changes in GABA‐A receptors following their prolonged exposure to benzodiazepines, drugs known to produce tolerance and dependence, and also to be abused. Therefore, we investigated the effects of chronic diazepam treatment on the recombinant aip2y2S GABA‐A receptors (the most common type of GABA‐A receptors found in the brain), stably expressed in human embryonic kidney (HEK) 293 cells. Aliquots of the cell membrane preparations were used in binding studies to compare the effects of different diazepam concentrations on the number and affinity of [3H]flunitrazepam binding sites and on their allosteric interactions with GABA recognition sites. The results demonstrated that long‐term exposure of cells to a high concentration of diazepam (50 [xM) enhanced the maximum number (Bmax) of [3H]flunitrazepam binding sites, without changing their affinity (Kd). This effect appears to be the consequence of chronic diazepam administration, since short‐term diazepam treatment (30 min.) has not affected the number of benzodiazepine binding sites. In contrast, a lower concentration of diazepam (1 [xM) failed to modify the parameters (Bmax and Kd) of [3H]flunitrazepam binding. However, diazepam applied in both concentrations (1 [xM and 50 [xM) produced functional uncoupling between GABA and benzodiazepine binding sites. The increased number of benzodiazepine binding sites as well as allosteric uncoupling, observed after long‐term treatment with 50 [xM diazepam, were reduced by gabazine and bicuculline (competitive antagonists of GABA binding sites), respectively, suggesting a role of GABA recognition sites in these effects. Nevertheless, co‐treatment with GABA has not potentiated the enhancement of [3H]flunitrazepam binding sites induced by 50 [xM diazepam. On the other hand, the combination of 1 [xM diazepam with GABA had an additive effect on the maximum number of benzodiazepine binding sites. The results suggested up‐regulation of benzodiazepine binding sites after prolonged treatment with diazepam, applied in a concentration that might be found in drug abusers. On the other hand, the observed decrease in allosteric coupling between benzodiazepine and GABA binding sites has been already found with significantly lower doses of diazepam. In addition to benzodiazepine binding sites, GABA recognition sites seem to be important for the observed effects of diazepam. The present study suggests that at least two separable processes could be occurring during the long‐term diazepam treatment. We assume that these results might be useful for understanding the effects of chronic high dose benzodiazepine use on the nervous system. PP31 ANALGESIC OVERUSE IS MORE COMMON IN DEPRESSIVE MIGRAINE PATIENTS Vuković V, Mikula I, Lovrenčić‐Huzjan A, Budišić M, Demarin V University Hospital "Sestre milosrdnice", Department of Neurology, Zagreb Background: Medication overuse is relatively common among patients with frequent headaches. The aim of our study was to determine the relationship between depression, number of days with headache per month and the number of used medications. Patients and methods: A total of 66 patients (54 women‐mean age 42 years and 8 men‐mean age 42 years) with migraine (without or with aura, MO, MA) or tension‐type headache (TTH) have been included into the study. MO , MA and TTH were diagnosed according to the ICHD‐2 criteria. All patients fulfilled the Beck depression score which classifies depression from 1‐13 as minimal (group A), 14‐19 as mild (group B), 2028 as moderate (group C) and 29‐63 as severe (group D). Days with headache and the number of analgesics and triptans was obtained from all patients.


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Results: There were 30 patients in group A, 17 in B, 10 in C and 9 in group D. The mean depression score was 7,4 ; 14,9; 23,1 and 42,1 respectively. There were 41 patients with MO, 10 with MA, 5 with TTH and 5 with MO/MA +TTH. The mean number of days with headache was 11,7 in group A; 11,2 in B;16 in C and 12,5 in D. The mean number of analgesics and triptans used was 24,8 in group A; 25,5 in B; 35,9 in C and 43, 5 in D. Although the number of days with headache per month has not significantly differed among groups, patients with moderate and severe depression take more medications for their headaches, P<0.05. Conclusions: Patients with higher depression score are more likely to use a higher number of acute medications ( analgesics nad triptans) for their headaches, although the number of days with headache is similar. Our results support earlier observations PP32 THE EFFECT OF SHORT‐TERM AND LONG‐TERM ZOLPIDEM TREATMENT ON RECOMBINANT α1β2γ2S SUBTYPE OF GABAA RECEPTORS Vlainić J, Jazvinšćak Jembrek M, Peričić D Laboratory for Molecular Neuropharmacology, Division of Molecular Medicine, Ruđer Bošković Institute, 10 000 Zagreb, Croatia Introduction: GABAA (γ-aminobutyric acidA) receptors are ligand‐gated ion channels composed of various polypeptide subunits. They possess binding sites for several drugs, including benzodiazepines. The aim of this study was to explore the mechanisms that underlie adaptive changes in GABAA receptors following their short‐term and long‐term exposure to hypnotic zolpidem, the non‐benzodiazepine agonist of benzodiazepine binding sites. Such studies might be important as the prescription of zolpidem is increasing and prolonged zolpidem treatment, as recently shown, induces tolerance (Vlainić and Peričić, Neuropharmacology, 56:1124‐30, 2009). Materials and methods: Human embryonic kidney (HEK 293) cells stably expressing recombinant α1β2γ2S GABAA receptors, the most common type of GABAA receptors found in the brain, were exposed to zolpidem (1 and 10 |iM ) for 2 h on one, two or three consecutive days or continuously for 48 h. Cell membranes were isolated 24 h after short‐term or immediately after long‐term treatment and radioligand binding studies were used to determine the parameters of [3H]flunitrazepam ([3H]FNZ) binding sites, as previously described (Peričić et al., Naunyn‐Schmiedeberg' s Arch. Pharmacol. 375: 177‐87, 2007). Membrane preparations were incubated with increasing concentrations of non‐radioactive flunitrazepam and a fixed concentration (1 nM) of [3H]FNZ) to determine the maximum number (Bmax) and dissociation constant (Kd) of benzodiazepine binding sites. In stimulation studies the ability of GABA to potentiate [3H]FNZ binding was used to assess the allosteric interactions between GABAA receptor binding sites. Results: A single (2 h) or repeated (2 or three days for 2 h daily) exposure of cells to zolpidem did not affect either the Bmax or the Kd value of [3H]FNZ binding sites. Addition of GABA enhanced [3H]FNZ binding to membranes obtained from control and zolpidem treated cells in a concentration‐dependent manner, but there was no difference in either the potency (EC50) or efficacy (Emax) of GABA to potentiate [3H]FNZ binding. On the contrary, prolonged occupation (48 h) of benzodiazepine binding sites by zolpidem (10 |iM) produced an up‐regulation (~150%) of [3H]FNZ binding sites with no change in their affinity, as well as the functional uncoupling between GABA and benzodiazepine binding sites, as evidenced by a decreased ability of GABA to stimulate [3H]FNZ binding. Conclusions: The results suggest that in our model, characterized by the presence of a single well defined receptor subtype, the continuous, but not intermittent short‐term exposure of cells to zolpidem (the latter being more similar to the therapy in humans), is able to induce adaptive changes in GABAA receptors possibly related to development of tolerance and dependence. These changes are not substantially different from those obtained after prolonged exposure of cells to high doses of classical benzodiazepines (Švob Štrac et al., Brain Res. 1246:29‐40, 2008).


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P4 MOLECULAR NEUROSCIENCE

PP33 IMMUNE RESPONSE AND THE MODULATION OF POST‐ISCHEMIC NEUROGENESIS Bohaček I1, Gorup D1, Lalancette‐Hebert M2, Weng YC2, Gajović S1, Križ J2 1Laboratory for Neurogenetics and Developmental Genetics, Croatian Institute for Brain Research , School of Medicine, University of Zagreb, Croatia 2Dept. of Psychiatry & Neuroscience, Faculty of Medicine, Laval University, Quebec, Canada Experimentally and clinically, brain response to ischemic injury is associated with an acute and prolonged inflammatory response characterized by the activation of resident glial cells, production of inflammatory cytokines and leukocyte and monocyte infiltration in the brain. Evidence suggests that these events may contribute to ischemic injury and may affect the processes associated with brain repair, such as neurogenesis. A current view is that injury induced neurogenesis may carry a significant impact on post‐ischemic brain plasticity and recovery. At present, the precise molecular mechanisms involved in induction and/or regulation of the post‐injury neurogenesis remain unknown. Mounting evidence however suggests that inflammation may play an important role. Therefore the aim of this study was to investigate the effects of microglial activation and the innate immune response, in particular the Toll like receptor 2 (Tlr‐2), on the modulation of the post‐ischemic neurogenesis. Our initial findings using Tlr‐2 reporter mouse have demonstrated that and an active pro‐inflammatory response may persist several months after stroke and may temporally coincide with the processes of the brain recovery. Unilateral transient focal cerebral ischemia was induced by intraluminal filament occlusion of the middle cerebral artery (MCA) during 1 hour followed by a different reperfusion periods. The surgery was carried out on 2‐3 months old wild‐type (WT) or TLR2 knockout male mice. To analyze acute and the long term response to ischemia, the animals were allowed to survive 24, 72 hours and 7, 14 and 30 days after initial stroke. The animals were transcardially perfused with 30 ml of 0.9% saline, followed by 4% paraformaldehyde (PFA). Tissue sample were then postfixed overnight in 4% PFA and equilibrated in PBS /30% sucrose for 48 hours, freeze at ‐20°C and cut into coronal section. The immunohistological analysis of the brain sections at different time points after stroke revealed that an early phase of inflammatory response was associated with a marked activation of microglial cells, Iba1 staining that co‐localized with Tlr2 immunoreactivity. The activated Iba1/Tlr2 positive cells were situated at the side of the ischemic lesion and the peri‐infarct zone. Contrary to a massive infiltration of the activated glial cells, at the site of the lesion at earlier time points we did not observe doublecortin (DCX) positive neuronal precursor cells. The analysis of the brain sections at later time points after stroke (14 and 30 days after) in addition to persistent microglial activation, revealed a stream of DCX positive cells migrating from the SVZ towards the site of ischemic lesion. Interestingly, a double immunfluorescence analysis revealed that some of the DCX positive cells were also immunoreactive for Tlr‐2. These results suggest an implication of the innate immune response in the modulation of post‐ischemic neurogenesis. PP34 GLYCOMICS OF BRAIN GLYCOSAMINOGLYCANS BY CHIP‐BASED ELECTROSPRAY IONIZATION ION TRAP MULTISTAGE MASS SPECTROMETRY Flangea Corinal,2, Sisu Eugen2, Seidler Daniela G3, Vukelić Željka4, Zamfir Alina Dl,5

1Mass Spectrometry Laboratory, National Institute for Research and Development in Electrochemistry and Condensed Matter, Timisoara, Romania; 2Department of Biochemistry, "Victor Babes" University of Medicine and Pharmacy, Timisoara, Romania; 3Institute for Physiological Chemistry and Pathobiochemistry, University of Munster, Munster, Germany; 4Department of Chemistry and Biochemistry, Faculty of Medicine, University of Zagreb, Zagreb, Croatia; 5Department of Chemical and Biological Sciences, "Aurel Vlaicu" University of Arad, Arad, Romania Chondroitin sulfate (CS) and dermatan sulfate (DS) glycosaminoglycans display variability of sulfation in their constituent disaccharide repeats during chain elongation. Since a large proportion of the


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extracellular matrix of the central nervous system (CNS) is composed of proteoglycans, CS/DS disaccharide degree and profile of sulfation play important roles in modulation of neuronal functions, brain development and some of its pathological states. To investigate the sulfation pattern of CS/DS structures expressed in CNS we developed a novel method based on an advanced system encompassing fully automated chip nanoelectrospray ionization (nanoESI) in the negative ion mode and high capacity ion trap (HCT) multistage mass spectrometry (MS2‐MS3) by collision induced dissociation (CID). This method, introduced here for the first time in glycomics of brain glycosaminoglycans, was particularly applied to structural investigation of disaccharides obtained by beta‐elimination and digestion with chondroitin B and AC I lyase of hybrid CS/DS chains from a wild type mouse brain. Screening in the chip‐MS mode of the DS disaccharide fraction, resulting after depolymerization with chondroitin B lyase, revealed molecular ions assigned to monosulfated disaccharide species having a composition of 4,5-δ-[IdoA‐GalNAc]. By optimized CID MS2‐MS3, fragment ions supporting the localization of sulfate ester group at C‐4 within GalNAc residue were produced. Chip eSi MS profiling of the CS disaccharide fraction, obtained by depolymerization of the same CS/DS chain using chondroitin AC I lyase, indicated the occurrence of mono‐ and bisulfated 4,5‐δ‐[GlcA‐GalNAc]. The site of oversulfation was determined by MS2‐MS3, which provided sequence patterns consistent with a rare GlcA‐3‐sulfate‐GalNAc‐6‐sulfate structural motif. PP35 THE ROLE OF 5HT‐RELATED GENES IN AUTISM: ASSOCIATION STUDY Hranilović Dubravka1, Novak Rudjer2, Babić Marina3, Ivanković Tomislav4, Matak Ivica5, Mokrović Gordana6, Blažević Sofia1, Štefulj Jasminka6 and Jernej Branimir6 1Department of Animal Physiology, Faculty of Science, University of Zagreb, 2Department of Biochemistry and Molecular Biology, Faculty of Pharmacy and Biochemistry, University of Zagreb, 3Center for Proteomics, Medical Faculty, University of Rijeka, 4Department of Botany, Faculty of Science, University of Zagreb, 5Department of Pharmacology, Medical Faculty, University of Zagreb, 6Department of Molecular Biology, Rudjer Bošković Institute, Zagreb Introduction: Autism is a neurodevelopmental syndrome, with onset in early childhood, characterized by social and communication dysfunction and stereotypic behaviors. Recent findings indicate that disturbances in serotonergic (5HT) neurotransmission represent one of the biological substrates of this disorder. Alterations in the expression of one or more 5HT‐related genes might lead to the dysregulation of 5HT transmission in the brain, affecting so its early development and resulting in autistic behavioral symptoms. We have studied the association of the following 5HT‐related genes: tryptophan hydroxylase 1 and 2 (Tphl, Tph2), monoamine oxydase A and B (MAOA, MAOB), 5HT transporter (5HTT) and 5HT2A receptor (5HT2Ar) with autism in the Croatian population. Materials and Methods: DNA was isolated from blood samples of 103 autistic and 368 control subjects and genotyped, using polymerase chain reaction and restriction analysis, for A218C Tphl, G703T Tph2, uVNTR MAO A, A/G MAOB, 5HTT‐LPR and ‐1438AG 5HT2Ar polymorphisms. Distributions of alleles and genotypes were compared between the autistic and control groups. Results: There were no significant differences in the distribution of alleles and genotypes of the Tph, MAO and 5 HTT polymorphisms between autistic and control subjects. However, a significant difference in the distribution of genotypes (p=0.014) and alleles (p=0.0054) of the 5HT2Ar polymorphism has been observed, with an increased incidence of G allele and GG genotype in the autistic group. In addition, an increase in the relative autistic to control subject ratio followed the increase in number of G alleles. The observed trend was significant (p=0.0035). Conclusion: The results provide evidence for the association of the 5HT2Ar gene with autism in the Croatian population and indicate a possible role of this 5HT‐related gene in susceptibility to autism spectrum disorders.


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PP36 GANGLIOSIDES OF ADULT HUMAN BRAIN PRECENTRAI AND POSTCENTRAL GYRUS: A CHIP‐MASS SPECTROMETRY BASED COMPOSITION AND STRUCTURE ANALYSIS Jeličić Joško1*, Ivković Vanja1*, Marinčić Dragana1*, Zamfir Alina D2,3, Vukelić Zeljka1 1Department of Chemistry and Biochemistry, Faculty of Medicine, University of Zagreb, Zagreb, Croatia; 2Mass Spectrometry Laboratory, National Institute for Research and Development in Electrochemistry and Condensed Matter, Timisoara, and 3Department of Chemistry and Biology, "Aurel Vlaicu" University of Arad, Arad, Romania As plasma‐membrane components, enriched in membrane microdomains (lipid rafts), and expressed as cell surface antigens, glycosphingolipids participate in vital processes and events of eukaryotic cells: cell‐to‐cell recognition/communication and cell signaling, growth, differentiation, apoptosis etc. Mammalian brain tissues, comparing to other tissue types, contain the highest concentrations of sialylated glycosphingolipids ‐ gangliosides. Specific changes in their expression occur during brain development, maturation, aging, and due to diseases. Therefore, gangliosides represent important biomarkers able to provide information upon various processes/events in CNS in health and pathology. In this contribution, we report to our knowledge the first data describing ganglioside compositions in two functionally defined regions of adult human brain (20 and 17 years): the precentral gyrus (the location of primary motor cortex) and the postcentral gyrus (the location of primary somatosensory cortex). The comparative study was conducted employing advanced chip‐mass spectrometry (MS) and high‐performance thin‐layer chromatography (HPTLC) methods. Dissected tissue samples were homogenized and total gangliosides were extracted and purified as native mixtures for analyses. HPTLC was used for both purposes: collection of compositional data following resorcinol‐HCl staining of separated ganglioside patterns and preparative isolation of separated fractions for subsequent MS analysis. The MS screening and sequencing analyses were performed on a High Capacity Ion Trap Ultra (PTM discovery) mass spectrometer (Bruker Daltonics, Bremen, Germany) coupled, via mounting system, with a NanoMate robot incorporating ESI 400 Chip technology (Advion BioSciences, Ithaca, USA). Tandem MS was carried out by collision‐induced dissociation. The resorcinol‐HCl‐stained HPTLC ganglioside patterns of both analyzed brain regions were basically similar, resembling the reference pattern of adult frontal cerebral cortex, but with slightly elevated domination of GMl‐migrating fraction, almost equal relative proportions of GD1a and GD1b fractions, and slightly higher proportions of GM3, GQ1b and GX fractions. Comparative MS screening of the analyzed native mixtures revealed highly complex ganglioside compositions with more than 100 distinct species. Based on peak intensities of detected molecular ions in the negative ion mode, characteristic differences in relative abundances of individual species in precental vs. postcental gyrus were observed. High variability of ceramide portions was detected for the majority of ganglioside forms defined by oligosaccharide moieties. Besides mono‐ to polysialylated forms, both regions contained structures with fucosyl‐ or additional GalNAc‐residue, as well as alkali‐labile O‐acetylated and lactonized gangliosides as minor components. Sequencing data allowing structural elucidation of some characteristic species will be presented. (*Equal contribution) PP37 EXPRESSION ANALYSIS OF STAM2 GENE IN THE BRAIN USING A GENE TRAP DERIVED TRANSGENIC MOUSE MODEL Kapuralin Katarina, Ćurlin Marija, Dobrivojević Marina, Srpak Nives, Mitrečić Dinko, Gajović Srećko Croatian Institute for Brain Research, School of Medicine University of Zagreb, Šalata12, Zagreb, Croatia STAM2 was identified as a phosphotyrosine protein involved in cellular response to variety of cytokines and growth factors. Together with HRS (hepatocyte growth factor regulated tyrosine kinase substrate) and Epsl5 (EGFR pathway substrate clone no. 15) it has been implicated in endosome mediated intracellular membrane trafficking. In order to investigate expression and function of STAM2, mouse carrying a gene trap modification of Stam2 gene was created. Gene trap method involves genetical modification of embryonic stem (ES) cells with a nonhomologous DNA vector


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containing a splice acceptor and fused promotorless lacZ and neoR genes. After a large scale screen of mutated ES cells, corresponding mouse lines were produced, and the line carrying modification of Stam2 gene, latter named Stam2GtlGaj, was selected for further investigation. As a consequence of gene trap modification, lacZ gene is in frame with Stam2, therefore expression pattern of Stam2 can be assessed by simple histochemical detection of beta‐galactosidase activity via its substrate X‐gal. This was used to determine Stam2 expression pattern in the mice carrying gene trap insertion. The advantage of this approach is that the expression is detected on protein level. To verify the above results in situ RNA hybridization using radioactive (S3 5) labelled probe was made. Stam2 is expressed in the central nervous system in the regions containing numerous neuron cell bodies, i.e. mainly in the cortex and the hippocampal principle cells layers, the habenula and the piriform cortex. Besides this, Stam2 signal appears to be expressed in a number of cells throughout several brain nuclei. As STAM2 together with HRS (hepatocyte growth factor regulated tyrosine kinase substrate) forms the ESCRT‐0 complex implicated in sorting of ubiquitinated receptors toward late endosomes/multivesicular bodies and subsequent degradation in the lysosome, the observed expression suggests the importance of STAM2 in the regulation of growth factor and cytokine signalling in the nervous system. PP38 CHARACTERIZATION OF GANGLIOSIDE COMPOSITION OF SELECTED FETAL HUMAN BRAIN REGIONS COMBINING CHIP‐BASED MASS SPECTROMETRY AND THIN‐LAYER CHROMATOGRAPHY Marinčić Dragana1, Zamfir Alina D.2,3, Kos Marina4, Vukelić Željka1 1Department of Chemistry and Biochemistry, Faculty of Medicine, University of Zagreb, Zagreb, Croatia; 2Mass Spectrometry Laboratory, National Institute for Research and Development in Electrochemistry and Condensed Matter, Timisoara, and 3Department of Chemistry and Biology, "Aurel Vlaicu" University of Arad, Arad, Romania; 4Department of Clinical Pathology »Ljudevit Jurak«, University Hospital »Sestre milosrdnice«, Zagreb, Croatia Gangliosides (GGs), a class of sialic acid‐containing glycosphingolipids, are constituents of external leaflet of plasma membranes in mammalian tissues; their highest content and pattern complexity are present in brain cells. Their compositions vary in different brain regions and characteristically change during ontogenesis; specific changes of the normal pattern have been observed in various neuropathological conditions. Certain GG species are involved in key brain developmental processes: neuritogenesis, cell motility, axon guidance, apoptosis, cell‐to‐cell recognition. Here, we represent data of compositional and structural characterization of native GG mixtures and fractions from 24 weeks of gestation fetal human brain neocortical regions that correspond to prospective precentral and postcentral gyrus (i.e. prospective primary motor and primary somatosensory cortex, respectively). The native GG mixtures were extracted and purified from the tissue homogenates. For analysis, our recently introduced mass spectrometry (MS) approaches were combined with high‐performance thin‐layer chromatography (HPTLC). GG fractions separated by HPTLC using suitable chloroform/methanol/0.2% aq. CaCl2 developing system(s) were visualized with resorcinol‐HCl reagent. Also, the GG fractions were isolated by preparative TLC. MS screening and sequencing analyses were performed on a High Capacity Ion Trap Ultra (PTM discovery) mass spectrometer (Bruker Daltonics, Bremen, Germany) coupled with a NanoMate robot incorporating ESI 400 Chip technology (Advion BioSciences, Ithaca, USA). Tandem MS was carried out by collision‐induced dissociation. HPTLC GG patterns of the analyzed fetal brain regions were similar, containing fractions migrating as GM4, GM3, GM2, nLM1, GM1, GD3, GD1a, GD2/GD1‐alpha/GT3, GD1b, GX, GT1b and GQ1b; the GDla‐migrating fraction was quantitatively dominant. By MS analysis it was distinguished that all HPTLC‐separated bands contained several distinct species. More than 140 species were detected in each GG mixture by MS1 screening in negative ion mode. Most of GG forms with defined oligosaccharide structure were represented by several species differing by the composition of ceramide portion. The relative abundance of individual species was moderately different in two analyzed regions. Higher number and higher total abundance of mono‐ and disialylated structures


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was found in the prospective postcentral comparing to the precentral gyrus. In contrast, the prospective precentral gyrus contained larger number of polysialylated (tri‐to even hexasialylated) structural variations. Species such as tetra‐ and pentasialo‐forms, O‐Ac‐GM3, di‐O‐Ac‐GD3 and O‐Ac‐GT3, and to a certain extent GM4, GT3, Fuc‐GT1 and GalNAc‐GT1 were considerably more abundant in both fetal brain regions than previously observed in adult human brain; this further confirms their categorization as fetal brain markers and developing brain‐associated species. PP39 LOCALIZATION OF DOPAMINE RECEPTORS IN A NEUROBLASTOMA CELL LINE Mladinov M1, Diana A2, Lam LT3, Man NT3, Holt I3, Morris GE3, Šimić G1

1Department of Neuroscience, Croatian Institute for Brain Research, Zagreb School of Medicine, Croatia; 2Department of Cytomorphology, University of Cagliari, Cagliari, Italy; 3Wolfson Centre for Inherited Neuromuscular Disease, TORCH Building, RJAH Orthopaedic INTRODUCTION: Human neuroblastoma cell line SH‐SY5Y is a dopaminergic neuronal cell line expressing tyrosin‐hydroxoylase, vesicular monoamine transporter VMAT2 and dopamine receptors (DR). It is a widely used model to study mechanisms of neurodegeneration, dopamine toxicity and DR ligands. However, there is no report about the expression and the subcellular localization of DR in these cells. MATERIALS AND METHODS: SH‐SY5Y cell line was terminally differentiated into neuron‐like cells after 5‐day treatment with retinoic acid (10 microM), followed by 3‐day treatment with brain‐derived neurotropic factor (BDNF, 50ng/mL). Cells were fixed with 4% paraformaldehyde and stained using novel anti‐DRD2, anti‐DRD3 and anti‐DRD4 monoclonal antibodies. The specificity of the antibodies used has been previously characterized by Wolstencroft et al. (2007). RESULTS: We found strong expression of DRD2 along the cell membrane, but also in cytoplasmatic vesicles. The expression of DRD3 was significantly weaker with almost no vesicle‐like reactivity. We observed no DRD4 immunoreactivity in this cell line. CONCLUSIONS: We showed an expected membrane localization, but also a significant endosomal localization of DRD2. The information on structure and function of endosomes that contain DRD2 are still lacking. We suppose they represent late endosomes and hope that future studies aiming to define their origin will help us understand the whole cycle of molecular events, from DRD2 cell membrane expression and internalization to their storage and reuse. PP40 D2, D3 AND D4 DOPAMINE RECEPTOR EXPRESSION IN THE PREFRONTAL CORTEX OF NORMAL AND SCHIZOPHRENIC BRAINS Mladinov M1, Mayer D2, Jovanov‐Milošević N1, Kostović I1, Šimić G1

1Department of Neuroscience, Croatian Institute for Brain Research, Medical School Zagreb, Croatia, 2Department of Legal Medicine, Medical School Zagreb, Croatia INTRODUCTION: The knowledge of the dopamine system is enormously important for understanding the human behavior as well as for the pathophysiology of many neurological and psychiatric diseases. Functional disturbances of the midbrain dopamine trajectories to the prefrontal cortex (PFC) have been commonly implicated to psychiatric disorders like schizophrenia, autism, depression and drug abuse. So far, the majority of studies addressing the localization of dopamine receptors (DR) in the human brain used crude autoradiographic approach of ligand binding or mRNA analysis, while the protein expression and precise anatomical localization of DR in PFC, particularly of the D2‐like group (DRD2, DRD3 and DRD4), have been largely neglected. SUBJECTS AND METHODS: Human brain tissue was taken during autopsy from 4 schizophrenic patients (age range: 51‐59 years ) and 3 age‐matched controls (age range: 48‐56 years) with no history of neurologic or psychiatric disorders. Three tissue specimens were collected from the right PFC of each brain: dorsolateral (BA9/BA46), orbitomedial ( BA11/BA12) and dorsomedial cortex (BA9). The blocks were fixed in 4% paraformaldehyde, dehydrated, paraffin‐embedded and cut in 12‐micron thick sections. For immunocytochemistry the sections were stained using novel anti‐DRD2,


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anti‐DRD3 and anti‐DRD4 antibodies. The specificity of the antibodies used has been previously characterized by Wolstencroft et al. (2007). RESULTS: We found strong DRD2 immunoreactivity in pyramidal neurons of layer III and moderate in layer V pyramidal cells in all three regions analyzed. Inversely, the expression of DRD4 was stronger in layer V pyramidal neurons. The expression of DRD3 was generally weaker than the expression of DRD2 and DRD4. Contrary to normal controls, a significant number of DRD2 ‐immunopositive reactive astroglial cells, predominantly in the subcortical white matter, were found in all four schizophrenic brains. This finding represented the major difference in DR expression between normal and schizophrenic brains. CONCLUSIONS: The finding that pyramidal cells of layers III and V express DR of the D2 ‐like group confirms that the activity of prefrontal neuronal circuits is strongly modulated by dopamine transmission. The strong expression of DRD2 receptors in reactive astrocytes of all four schizophrenic brains analyzed suggests its possible importance for the pathogenesis of schizophrenia. NOTE: This poster was presented at the symposium "Neurogenomics and neuroimaging of developmental disorders" held from 30 April ‐5 May 2009 in Dubrovnik. PP41 GENE EXPRESSION PROFILING OF CEREBELLAR TISSUE OF GANGLIOSIDE DEFICIENT MICE Mlinac Kristina1, Režen Tadeja2, Heffer Marija3, Rozman Damjana2, Kalanj Bognar Svjetlana1 1Croatian Institute for Brain Research, School ofMedicine, University ofZagreb; 2Centre for Functional Genomics and Biochips, School ofMedicine, University ofLjubljana; 3Department of Biology, School ofMedicine, University of Osijek Gangliosides, glycosphingolipids containing sialic acids, are localized in the outer leaflet of the plasma membranes. Together with cholesterol, gangliosides are organized in specialized membrane microdomains (lipid rafts) involved in membrane‐associated events such as cell‐cell interaction, adhesion, cell differentiation, growth control and receptor functions. Greatest variety and amount of ganglioside structures have been found in brain tissue. Gangliosides are involved in brain development and maturation; on the other hand, deficiency of ganglioside synthesis leads to degeneration and altered axon‐glial interactions in central nervous system. In this study we analyzed the differences between transcriptomes of GD3 synthase (Siat8a) knockout (KO) and wild type (wt) mice. Siat8a knockout mice lack disialylated b‐series ganglioside structures (GD3, GD2, GDlb, GTlb). Upon aging, these mice show dysmyelination and impaired nerve regeneration. GTlb ganglioside is a ligand for myelin‐associated glycoprotein (MAG) and mutations in the human ortholog of Siat8a gene (ST8SIA1) have recently been linked with the risk of developing multiple sclerosis. RNA was isolated from brain samples of 3 wt and 3 KO animals. After quantitative and qualitative analysis of the RNA, cerebellar samples were labeled for two color microarray experiments. Samples were hybridized to Agilent's 1x22K Whole Mouse Genome Oligo Microarrays (6 arrays total) and analyzed with GenePix Pro and BRB‐Assay Tools. Gene annotation was performed using publicly available annotation tools ‐ GeneCoDis2 and DAVID Bioinformatics Database Gene ID Conversion Tool. After extensive analysis (and consideration of feature exclusion criteria) a list of differentially expressed genes was retrieved. The gene found to be most upregulated was Kras (v‐Ki‐ras2 Kirsten rat sarcoma viral oncogene homolog) involved in Ras signaling pathway; one of the most downregulated genes found was Mapkapk2 (MAP kinase‐activated protein kinase 2). In the biological process ontology the most frequent annotation term was signal transduction. Preliminary results indicate that the majority of differentially expressed genes in the cerebellum of GD3 synthase deficient mice are involved in the signal transduction. In order to confirm these results, the expression of individual genes needs to be determined. We find the results very interesting, having in mind that normal cerebellar tissue is particularly enriched in b‐series gangliosides and that in vitro studies showed gangliosides to be implicated in Ras signaling pathway. Our results show for the first time in animal model that deficient synthesis of GD3 and consequently other b‐series gangliosides, beside effects on membrane lipid homeostasis, is also probably associated to alterations in complex signaling pathways.


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PP42 SEROTONIN 5‐HT2A RECEPTOR GENE POLYMORPHISM IN POSTTRAUMATIC STRESS DISORDER Muck‐Šeler D1, Babić A1, Pivac N1, Nedić G1, Mustapić M1, Kozarić‐Kovačić D2

1Division of Molecular Medicine, R.Bosković Institute, Zagreb, Croatia, 2University Hospital Dubrava, Department of Psychiatry, Referral Centre for the Stress‐Related Disorders, Zagreb, Croatia Introduction: Posttraumatic stress disorder (PTSD) is a complex psychiatric and polygenetic disorder that occurs in some people exposed to extreme traumatic events. Literature data suggest that the alterations in serotonergic system and its receptors are involved in the aetiology and treatment of PTSD. Serotonergic receptors type 2A (5‐HT2AR) are widely distributed in the brain and periphery. The association between T102C polymorphism of 5‐HT2AR gene and personality traits including aggression and suicidality was reported. The aim of the present study was to determine a) the distribution of genotypes and alleles of the 5‐HT2AR 102T/C SNP in healthy Croatian population and b) the association between 5‐HT2AR gene polymorphism and vulnerability to PTSD in Croatian combat exposed war veterans. Subjects and Methods: The study included 264 war veterans (mean age ± SD, 39.0± 9.8 years), with current chronic PTSD (SCID for DSM‐IV), 89 combat exposed war veterans (age 38.1 ± 4.2 years) without PTSD and 291 healthy male persons (age 41.5 ± 12.1 years). PTSD diagnosis was evaluated with Structured Clinical Interview for DSM‐IV. Genomic DNA was extracted from whole blood by a salting out procedure. The DNA samples were genotyped using ABI Prism 7000 Sequencing Detection System apparatus using Taqman‐based allele‐specific polymerase chain reaction assay. The chi‐square (x2) test was applied to test the difference in genotype and allele frequencies of 5‐HT2AR polymorphism between groups. Results: There was no significant deviation from the Hardy‐Weinberg distribution for any group. Among healthy subjects CC, CT, TT genotype was found in 33.9%, 45.6% and 20.5% subjects respectively. CC genotype was found in 29.5%, CT in 43.2% and TT in 27.3% of war veterans with PTSD. Within war veterans without PTSD 38.2% carried CC, 46.1 CT and 15.7% TT genotype. 5‐HT2AR genotype frequencies were similarly (%2=1.05, df=2, p=0.591) distributed between the two groups of war veterans A statistically significant (x2=5.06, df=1, p=0.024) difference was observed in alleles distribution between veterans with and without PTSD. C allele carriers were 51.1% and 61.2%, T allele carriers were 48.9% and 38.8% in war veterans with and without PTSD, respectively. Conclusions: Our results suggest that T allele of the 5‐HT2AR 102T/C could be associated with the development of PTSD in subjects exposed to trauma. PP43 ASSOCIATION STUDY OF A FUNCTIONAL CATECHOL‐O‐METHYLTRANSFERASE POLYMORPHISM AND COGNITIVE FUNCTION IN PATIENTS WITH DEMENTIA AND MILD COGNITIVE IMPAIRMENT Nedić Gordana1, Borovečki Fran2, Klepac Nataša3, Mubrin Zdenko3, Hajnšek Sanja3, Muck‐Šeler Dorotea1, Pivac Nela1 1Rudjer Bošković Institute, Division of Molecular Medicine, Zagreb, 2Medical school, Center for Functional Genomics, Zagreb, 3University Hospital Center, Clinical hospital Centre Zagreb, Department of Neurology, Zagreb Introduction: A functional catechol‐o‐methyltransferase (COMT Vall58/108Met) polymorphism, A to G substitution, has been associated with cognitive processing in the normal brain, older age, mild cognitive impairment (MCI), various dementias and cognitive disruptions in schizophrenia. COMT is involved in the breakdown of dopamine and other catecholamines, especially in the frontal cortex; hence the carriers of Met allele, with the lower enzymatic activity, are expected to perform better on particular neurocognitive tests. Methods: Peripheral blood samples were collected from 40 dementia and 13 MCI patients. The determination of the neurological status of the dementia patients utilized the MMSE, NPI, ADAS‐COG, CDT, World pairs learning and recall/ Picture pairs learning and recall test and Visual attention test performed by an experienced neurologist. DNA from blood was extracted using the DNeasy Blood and Tissue Kit (Qiagen). COMT genotyping was performed by the TaqMan (Applied Biosystems) SNP analysis. The results, expressed as means ± SD, were evaluated using one‐way ANOVA followed


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by the Tukey's test. The Hardy‐Weinberg analysis was used to test the equilibrium of the population. The differences in the genotype and allele frequencies were evaluated using a χ2 test. Results: Patients with dementia and MCI differed significantly (p=0.035‐0.001, ANOVA) in age, duration of disease, MMSE, modified MMSE, ADAS, NPI, NPI apathy and CDT scores. The observed genotype distribution in patients with dementia (χ2=0.14; d.f.=l; P=0.933) and MCI (χ2=0.00; d.f.=l; P=1.000) did not differ significantly from the expected Hardy‐Weinberg equilibrium. In patients with dementia, but not with MCI, significant (p=0.016‐0.041, ANOVA) genotype‐induced differences were found in scores for MMSE, modified MMSE, VAT duration of numbers test, VAT time of response to numbers test, VAT average response to numbers test and WPLCR/PPLR unanswered. Carriers of Met/Met genotype had significantly (p=0.015‐0.037, Tukey's test) lower scores of MMSE or modified MMSE, significantly longer time to respond to VAT duration of numbers test, VAT time of response to numbers test and VAT average response to numbers test, and significantly greater number of unanswered questions to WPLCR/PPLR when compared to Met/Val or Val/Val genotypes. Conclusion: Our preliminary data showed significantly impaired performance in several neurocognitive tests in carriers of Met/Met genotype in patients with dementia, but not in patients with MCI, compared to either Met/Val or Val/Val genotype carriers. Although Met/Met genotype with more dopamine available in the frontal cortex should be associated with better neurocognitive test results than Met/Val or Val/Val genotype, our data on demented patients did not confirm this hypothesis. Further study on larger sample of patients is needed to clarify the role of COMT polymorphism in cognitive functions. PP44 REPORT ON MUTATION IN EXON 15 OF THE APC GENE IN A CASE OF BRAIN METASTASIS Pećina‐Šlaus Nives1,2, Majić Željka1, Musani Vesna3, Zeljko Martina1, Čupić Hrvoje4 1Laboratory of Neurooncology, Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Šalata 12, 10 000 Zagreb, Croatia, 2Department of Biology, School of Medicine, University of Zagreb, Šalata 3, 10 000 Zagreb,Croatia, 3Division of Molecular Medicine, Laboratory for Hereditary Cancer, Ruđer Bošković Institute, Bijenička c.,10 000 Zagreb, Croatia, 4Ljudevit Jurak Department of Pathology, University Hospital "Sisters of Mercy", Vinogradska 29, 10 000 Zagreb, Croatia Introduction: This study analyses exon 15 of the adenomatous polyposis coli gene (APC) in a 49‐year‐old male patient with brain metastasis (carcinoma metastaticum cerebri). Exon 15 comprises more than 75% of the coding sequence of the APC gene and is the most common target for both germline and somatic mutation. The involvement of APC as a general tumor suppressor gene in great variety of human tumors has been known for a long time. Materials and methods: The presence of somatic mutation was identified by PCR method and direct DNA sequencing of the metastasis and autologous lymphocyte samples. The exon 15 of APC gene was also investigated on the basis of RFLP of the PCR products, since the mutation was observed in an Msp I polymorphic site. Immunohistochemistry was performed in order to determine the presence of APC protein in metastasis sample. Results: The results of DNA sequencing of the PCR product from the metastasis revealed a G to A change at position 5883, codon 1961. The mutation was confirmed by RFLP analysis using Msp I endonuclease, since the mutation strikes Msp I restriction site. Immunohistochemical analysis revealed the lack of protein expression of this tumor suppressor gene. The main molecular activator of the wnt pathway, beta‐catenin, was expressed and located in the nucleus. Conclusion: The mutation is a silent mutation that might have consequences in creation of a new splice site. Different single‐base substitutions in APC exons need not to be evaluated only by the predicted change in amino acid sequence but rather at the nucleotide level itself. In our opinion such silent mutations should also be incorporated in mutation detection rate and validation.


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PP45 AXIN‐1 PROTEIN EXPRESSION AND LOCALIZATION IN GLIOBLASTOMA Pećina‐Šlaus Nives1,2, Nikuševa Martić Tamaral,2, Kokotović Tomislav1, Kušec Vesna3, Tomas Davor4 and Hrašćan Reno5 1Laboratory of Neurooncology, Croatian Institute for Brain Research, School of Medicine University of Zagreb, Šalata 12, HR‐10000 Zagreb, Croatia; 2Department of Biology, School of Medicine, University of Zagreb, Šalata 3, HR‐10000 Zagreb, Croatia; 3Clinical Institute of Laboratory Diagnosis, Clinical Hospital Centre Zagreb, Kišpatićeva 12, 10 000 Zagreb, Croatia; 4Ljudevit Jurak Department of Pathology, University Hospital "Sisters of Mercy", Vinogradska 29, 10000 Zagreb, Croatia, 5Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, HR‐10000 Zagreb, Croatia. Introduction: The etiology and pathogenesis of tumors of the central nervous system are still inadequately explained. The so called classical canonical wnt pathway, mediated through beta‐catenin, plays an essential role in development of many cancers. In the present study the expression patterns of a critical molecular component of wnt signaling pathway ‐ axin 1 was investigated in glioblastoma, the most aggressive form of glial tumors. Materials and methods: Immunostaining and image analysis revealed the quantity and localization of the axin 1 protein in 42 patients with glioblastoma. Results: Downregulation of this tumor suppressor protein expression was observed in 31% of tumors when compared to the levels of axin in healthy brain tissues. Axin was observed in the cytoplasm in 69% of glioblastoma samples, in 21.4% in both the cytoplasm and nucleus and 9.5% had expression solely in the nucleus. Mean values of relative axin's expression obtained by image analysis showed that the highest relative quantity of axin was measured when the protein was in the nucleus and the lowest relative quantity of axin when the protein was localized in the cytoplasm. Investigation on axin's existence at the subcellular level in glioblastomas suggests that axin's expression and spatial regulation is a dynamic process. Conclusions: Despite increasing knowledge on glioma biology and genetics, the prognostic tools for glioblastoma still need improvement. Our findings on expression of axin 1 may contribute to better understanding of glioblastoma molecular profile and could be used as prognostic marker of disease evolution and progression. PP46 LIVER AND PANCREAS GLYCOSPHINGOLIPID PHENOTYPES OF THE NEUROLOGICAL DIFFERENT RAT STRAINS Rešić Jasminka, Čikeš‐Čulić Vedrana, Zemunik Tatijana, Markotić Anita Department of Medical Chemistry and Biochemistry, University of Split, School of Medicine, Šoltanska 2, 21000 Split, Croatia Among three commonly used strains of laboratory rats, Wistar rats performe more neurological tasks better then Lewis and Sprague‐Dawley (SD) rats. Liver is the main site of insulin‐like growth factor (IGF) production and pancreas is the exclusive site of insulin production. Insulin stimulates neuronal development and IGF plays a particular role in the trophic maintenance of neurons involved in the coordination of sensorimotor function in the cerebellum. Glycosphingolipids (GSLs) are important mediators of insulin secretion and action. Therefore, this study investigated GSL phenotypes of liver and pancreas with hypothesis that they are different in three rat strains Total glycosphingolipid fractions (neutral and acidic) were analysed by high performance thin‐layer chromatography (HPTLC). Complex gangliosides were detected by HPTLC immunostaining using cholera toxin B subunit after neuraminidase pretreatment. Wistar rats had the highest, statistically significantly different, liver weight/body weight ratio. Major GSLs in the liver were globotetraosylceramide (Gb4), and monohexaosylceramide, equally expressed in all rat strains. Ganglioside GM3 was more expressed in the Wistar compared to Lewis and SD rats. There was no difference in complex ganglioside (GDI a, GDlb and GTlb) content in the liver of Wistar and Lewis rats, while SD rats showed only traces of complex gangliosides in the liver. SD rats expressed only traces of GDlb ganglioside in pancreas compared to higher GDlb expression in the


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pancreas of Wistar and Lewis rats. This study represent important differences in the liver and pancreas GSL phenotypes of three rat strains. These phenotypes have to be considered by the planning of neurochemical investigations of biochemical events linked to GSL expression.

P5 NEURODEGENERATIVE NEUROSCIENCE

PP47 EFFECT OF ENVIRONMENTAL ENRICHMENT ON MORPHOLOGY OF DENTATE GRANULE CELLS AND DEEP LAYER III AND LAYER V PYRAMIDAL CELLS OF OCCIPITAL CORTEX IN OLDEST‐OLD RAT Darmopil Sanja, Petanjek Zdravko, Mohammed Abdul B, Bogdanović Nenad Department of Neuroscience, Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Šalata 12, 10000 Zagreb, Croatia The hippocampus of aged rats shows marked age‐related morphological changes that could cause memory deficits. Experimental evidence has established that environmental enrichment attenuates memory deficits in aged rats. We therefore studied whether environmental enrichment produces morphological changes on the dentate granule cells of aged rats and deep layer III and layer V pyramidal neurons in the associate rat neocortex. Fifteen male Sprague‐Dawley rats, 24 months of age, were randomly distributed in two groups that were housed under standard (n=7) or enriched (n=8) environmental conditions for 26 days. Quantitative data of dendritic morphology and spine density were obtained on Golgi‐Cox stained sections. The environmental enrichment induced marked increase in the complexity and size of granule cells dendritic tree (total number of segments increased by 61% and length by 116%), and spine density (88% increase). Those values were much lower in both groups than in adult rat. It is possible that the dendrites had started to express regressive changes, a phenomenon that has been observed in the "oldest old" stage. This might imply that the effect of enriched environment is actually in slowing down the speed of regression. In the occipital cortex effects of the enriched environment were less pronounced and the values for total dendritic length and spine density were close to values found in the adult rats. For the layer III pyramidal cells increase of total segment number (37,5%) and intermediate segments dendritic diameter (20‐27%) was observed. Moderate effect were also observed for total dendritic length (19% increase) and spine number (30% increase). For the layer V pyramidal cells significant differences were found for the number of primary dendrites per neuron (30%) and diameter of intermediate segments (26%). There were large interindividual differences within the enriched group, indicating differential individual responses to environmental stimulation. Previous studies have observed changes produced by environmental enrichment in the morphology of dentate gyrus granule cells only in young and adult animals. The results of the present study show that environmental enrichment can also produce marked changes in dentate granule cell morphology in the senescent brain suggesting that hippocampus retains its neuroplastic capacity during aging. Interestingly, in the occipital cortex environmental enrichment induced only moderate changes. Our results suggest that in aging enriched environmental housing conditions are especially beneficial in the regions that undergo age‐related degenerative changes, attenuating age‐related dendritic regression and synaptic loss, thus preserving memory functions. This demonstrates the importance of cognitive stimulation for arresting regression in the hippocampus during normal aging and, moreover, for diminishing the deleterious effects of neurodegenerative disorders (i.e. Alzheimer's disease) on the hippocampus.


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PP48 ASSOCIATIONS OF THE DBH GENE WITH PLASMA DOPAMINE β‐HYDROXYLASE ACTIVITY IN ALZHEIMER'S DISEASE Mustapić M1, Presečki P2, Mimica N34, Pivac N1, Folnegović‐Šmalc V34, Muck‐ Šeler D1

1Division of Molecular Medicine R.Bosković, Institute, HR‐10000 Zagreb, Croatia, 2Psychiatric Hospital Sveti Ivan, Jankomir 11, HR‐10090 Zagreb, Croatia, 3Psychiatric Hospital Vrapče, University Department of Psychiatry, Bolnička cesta 32, HR‐10090. Zagreb. Croatia Introduction: Alzheimer's disease (AD) is complex and polygenic disorder. Polymorphisms within the dopamine β‐hydroxylase (DBH) gene could contribute to etiology of Alzheimer's disease (AD), given the well‐documented changes in the catecholamine‐mediated neurotransmission that occurs in this disorder. The aim of the present study was to investigate two DBH gene polymorphisms (rs1611115 and rs6271) and plasma DBH activity in patients with AD and healthy controls. Materials and Methods: The study included 254 patients (mean± SD age 78.1 ± 10.6 years; MMSE = 13.9 ± 11.9) with AD (NINCDS‐ADRDA and DSM‐IV‐TR criteria) and 40 age‐matched healthy controls (74.2 ± 8.8 years). Plasma DBH activity was determined by a photometric method by Nagatsu and Udenfriend and DBH genotype with TaqMan Real‐time allelic discrimination technique after extraction of DNA from whole blood with salting out procedure. Results: A significantly (F= 19.1; df= 1,292; p=0.0002; ANOVA) lower plasma DBH activity was found in AD patients (9.38±6.15 R7/1) compared to healthy controls (16.19±11.95 IU/1). There was a significant difference in plasma DBH activity (F=22.5; df=2,291; p<0.0001) between different genotypes. Distribution of genotypes and alleles of DBH polymorphism rs 1611115 did not differ

significantly between AD and control samples (χ‐square=0.12, df=2, p=0.941 and χ‐square=0.01, df=l, p=0.920, respectively). There were also, no significant differences in genotype and allele

distributions of the other DBH polymorphism rs6271 (χ‐square=2.56, df=2, p=0.278 and χ‐square=0.61, df=l, p=0.435) between groups. Two tested polymorphisms of DBH were not in the

linkage disequilibrium (χ‐square= 2.45, df=l, p =0.117). Interaction tested between the two independent factors (diagnosis x genotype) gave no significant (F=0.70; df=2,288; p=0.499) difference in plasma DBH activity. Conclusions: Our study did not provide evidence for association of two DBH genetic polymorphisms with AD. The results show significantly lower DBH activity in AD and therefore genotype‐controlled measurement of plasma DBH activity might be used as a potential biological marker in AD. PP49 PRELIMINARY STUDY OF THE COGNITIVE FUNCTION AND VAL66MET POLYMORPHISM IN THE BRAIN‐DERIVED NEUROTROPHIC FACTOR GENE IN PATIENTS WITH DEMENTIA AND MILD COGNITIVE IMPAIRMENT Pivac N1, Nedić G1, Borovečki F2,3, Klepac N2, Mubrin Z2, Hajnšek S2, Nikolac M1, Muck‐Šeler D1

1Division of Molecular Medicine, Rudjer Bošković Institute, POBox 180, HR‐10002 Zagreb, Croatia, 2Department of Neurology, Clinical Hospital Centre Zagreb, University Hospital Center, Kišpatićeva 12, HR‐1000 Zagreb, Croatia, 3Center for Functional Genomics, Zagreb Medical School, University of Zagreb, Šalata 2, HR‐10000 Zagreb, Croatia Introduction: Brain derived neurotrophic factor (BDNF) plays an important role in neuronal survival, differentiation, and synaptic plasticity in the brain. Reduced BDNF levels are found in dementia and cognitive decline. BDNF is controlled by a functional polymorphism, which consists of a substitution of valine (Val) into methionine (Met). This BDNF val66met polymorphism has been implicated in lower depolarization‐induced production of BDNF, lower n‐acetyl asparatate content, and reduced hippocampal activation during memory processing. The Met allele has been associated with reduced delayed episodic memory or working memory performance, but also with enhanced verbal reasoning ability. Aim: The hypothesis was that BDNF genetic variants may be associated with cognitive function in patients with dementia and mild cognitive impairment (MCI). Methods: A Val66Met polymorphism of the BDNF‐gene was studied in 40 patients with dementia and in 13 patients with mild cognitive impairment (MCI). The association of BDNF val66met with


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neurological and cognitive status in patients with dementia and MCI was examined using the MMSE, NPI, ADAS‐COG, CDT, World pairs learning and recall/ Picture pairs learning and recall test and Visual attention tests. DNA from blood was extracted using the DNeasy Blood and Tissue Kit (Qiagen). In DNA samples BDNF polymorphism was genotyped by a TaqMan (Applied Biosystems) analysis. Results were evaluated using one‐way ANOVA followed by the Tukey's test. The Hardy‐Weinberg analysis was used to test the equilibrium of the population. The differences in the genotype and

allele frequencies were evaluated using a χ2 test. Results: The genotype and allele frequencies for Val66Met polymorphism did not differ significantly between control subjects, and patients with dementia or MCI. Patients with dementia and MCI differed significantly (p=0.035‐0.001, ANOVA) in age, duration of disease, MMSE, modified MMSE, ADAS, NPI, NPI apathy and CDT scores. When subdivided according to the BDNF variants, patients with dementia differed significantly (F=8.422; p=0.012) only in the NPI disinhibition scores, while patients with MCI had significantly different (F=4.997; p=0.047) scores on World pairs learning and recall/ Picture pairs learning and recall test (time of un‐correct answers). The observed genotype

distribution in patients with dementia (χ2=0.97) and MCI (χ2=0.69) did not differ significantly (P>0.05) from the expected Hardy‐Weinberg equilibrium. Conclusion: The results from this preliminary study, showing that carriers of Val/Val genotype had better NPI disinhibition scores in dementia and shorter time of incorrect answers in MCI, are in line with the lower BDNF activity in Met/Met carriers. Although homozygosity for the Val allele was reported to confer an increased risk for dementia, no significant differences were found in the frequency of BDNF variants among patients or between patients and control subjects. PP50 OLIVE‐OIL ENRICHED DIET AND CHANGES IN TOTAL LIPIDS CONTENT OF BRAIN FROM MICE IN THE EARLY PHASE OF LIVER REGENERATION Pantović Radojka, Milin Čedomila Department of Chemistry and Biochemistry, Faculty of Medicine, University of Rijeka, Braće Branchetta 20, HR‐51000 Rijeka, Croatia Olive oil is an excellent source of monounsaturated fatty acids, and many known antioxidants, therefore it has a protective role on cells. In our experiments, we wanted to monitor changes in total lipids composition of cell membranes in the brain of mice fed with specific oils, and the role of herbal oils on regeneration and reparation of tissue. Aim: The aim of this study was to research the effects of olive oil enriched diet as well as the effects of liver regeneration after partial hepatectomy (pHx) on the total lipids content in the brain tissue of mice. Materials and methods: C57Bl mice were fed three weeks with standard food and food enriched with 5% of olive oil. Those fed with standard food were used as a control group. After three weeks animals were subjected to pHx, and 6th, 12th, 24th, and 48th hours after the operation they were sacrificed and their brains were taken out. Total lipids were extracted from brain tissue samples with a chloroform‐methanol (2:1, v/v) mixture according to Folch et al. Lipids were protected against oxidation by adding BHT to the solvents. The average amount of total lipids from brain was gravimetrically determined. Results and conclusions: Compared to the control group, the total lipids composition of brain in animals with pHx was reduced, therefore we can conclude that this operation, and the removal of 1/3 of the liver of mice fed with standard food has affected the mass content of brain lipids. In mice fed with olive oil enriched diet, the mass content of brain lipids was higher than that in the control group, so we can conclude that olive oil enriched diet has affected the total lipids composition of brain in mice. Our results show that olive oil has a protective role on the brain.


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PP51 ENVIROMENTAL ENRICHMENT HAS BENEFICIAL EFFECT ON COGNITION IN THE RAT MODEL OF SPORADIC ALZHEIMER’S DISEASE Osmanović J1, Šalković‐Petrišić M1, Riederer P2 1Department of Pharmacology and Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Šalata 11, HR 10 000 Zagreb, Croatia, 2Department of Clinical Neurochemistry, University Department of Psychiatry and Psychotherapy, University of Würzburg, Fuechsleinstr. 15, 97080 Würzburg, Germany Background: A growing body of evidence indicates indeed that in sporadic type of AD (sAD) physical and mental activities have beneficial effects on cognitive impairments, but long lasting cognitive and physical stimulation has not been studied yet in patients with sAD. We investigated whether the long‐term enriched housing and intensive cognitive training could have a positive effect on cognitive deficit in learning and memory found in streptozotocin‐intracerebroventricularly (STZ‐icv), treated rats, as an experimental model of sAD model. Material and Methods: Adult male Wistar rats were treated icv with streptozotocin while the controls received vehicle only. Half of the control and STZ‐icv treated group has been afterwards rendered to enriched housing (EH) and intensive mental training in the Morris Water Maze Swimming test and Dray maze for three months. Data were statistically analyzed by Mann‐Whitney U‐test (p<0.05). Results: Significant cognitive deficit has been found in Morris Water Maze Swimming test (MWM) in the third week (29%) which was not significant after the EH and intensive training, three months following the STZ‐icv treatment in comparison to the controls. Also, in MWM test the increase in AUC (area under the curve) during the time of learning in STZ‐icv group was statistically significant only before the EH and intensive training, showing the slower learning process in STZ‐icv rats. Four weeks after STZ‐icv treatment, the significant increase in anxiety (142%) and decrease in moving (31%) were found in dry maze while no change was found in the same test after the EH and intensive training three months following the STZ‐icv treatment. Conclusion: Our results show that early and long term intensive psychical and mental training has beneficial affect on the memory loss and anxiety caused by the STZ‐icv treatment. Supported by Croatian MZOS (108‐1080003‐0020) and DAAD

STUDENT ORAL PRESENTATIONS OP1 INFLUENCE OF SHAPE ON LIKENESS OF OBJECTS Ugrina Marija Eva University of Zadar, Department of Psychology People constantly make snap judgments about objects encountered in the environment. Such rapid judgments are based on the physical properties of the targets. Liking of visual objects has been shown to be affected by factors such as symmetry, prototypicality, contrast, complexity, and perceptual fluency. The idea for this experiment is based on the fact that there is one more visual primitive that mediates the formation of such rapid impressions ‐ contour of an object. We hypothesized that sharp transitions in contour might convey a sense of threat, and therefore trigger a negative bias. Also, people may tend to react faster to such objects, for which an explanation could be found in evolutionary psychology ‐ sharp objects could be perceived as dangerous and reaction time for dangerous objects is inherently faster. We also hypothesized that people prefer familiar objects because their experience with such objects made them familiar with the way they should react to those objects. For the purpose of this experiment a program was developed in which participants had to evaluate the likeness of an object, at first by forced choice and then later on the Likeri type scale. Also, the reaction time was measured. Objects differentiated on two dimensions ‐ curviness and familiarity. Results have shown that type of contour of a visual object and its familiarity influences people's


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attitudes toward that object in a way that participants preferred curved shaped objects to sharp ones and their reaction time was shorter for sharp objects. Still, the hypothesis considering the preference of known objects over the abstract ones was just partially confirmed. OP2 GENE EXPRESSION PROFILING OF CEREBELLAR TISSUE OF GANGLIOSIDE DEFICIENT MICE Mlinac Kristinal, Režen Tadeja2, Heffer Marija3, Rozman Damjana2, Kalanj Bognar Svjetlanal lCroatian Institute for Brain Research, School of Medicine, University of Zagreb; 2Centre for Functional Genomics and Biochips, School of Medicine, University of Ljubljana; 3Department of Biology, School of Medicine, University of Osijek Gangliosides, glycosphingolipids containing sialic acids, are localized in the outer leaflet of the plasma membranes. Together with cholesterol, gangliosides are organized in specialized membrane microdomains (lipid rafts) involved in membrane‐associated events such as cell‐cell interaction, adhesion, cell differentiation, growth control and receptor functions. Greatest variety and amount of ganglioside structures have been found in brain tissue. Gangliosides are involved in brain development and maturation; on the other hand, deficiency of ganglioside synthesis leads to degeneration and altered axon‐glial interactions in central nervous system. In this study we analyzed the differences between transcriptomes of GD3 synthase (Siat8a) knockout (KO) and wild type (wt) mice. Siat8a knockout mice lack disialylated b‐series ganglioside structures (GD3, GD2, GDlb, GTlb). Upon aging, these mice show dysmyelination and impaired nerve regeneration. GTlb ganglioside is a ligand for myelin‐associated glycoprotein (MAG) and mutations in the human ortholog of Siat8a gene (ST8SIA1) have recently been linked with the risk of developing multiple sclerosis. RNA was isolated from brain samples of 3 wt and 3 KO animals. After quantitative and qualitative analysis of the RNA, cerebellar samples were labeled for two color microarray experiments. Samples were hybridized to Agilent’s 1x22K Whole Mouse Genome Oligo Microarrays (6 arrays total) and analyzed with GenePix Pro and BRB‐Assay Tools. Gene annotation was performed using publicly available annotation tools ‐ GeneCoDis2 and DAVID Bioinformatics Database Gene ID Conversion Tool. After extensive analysis (and consideration of feature exclusion criteria) a list of differentially expressed genes was retrieved. The gene found to be most upregulated was Kras (v‐Ki‐ras2 Kirsten rat sarcoma viral oncogene homolog) involved in Ras signaling pathway; one of the most downregulated genes found was Mapkapk2 (MAP kinase‐activated protein kinase 2). In the biological process ontology the most frequent annotation term was signal transduction. Preliminary results indicate that the majority of differentially expressed genes in the cerebellum of GD3 synthase deficient mice are involved in the signal transduction. In order to confirm these results, the expression of individual genes needs to be determined. We find the results very interesting, having in mind that normal cerebellar tissue is particularly enriched in b‐series gangliosides and that in vitro studies showed gangliosides to be implicated in Ras signaling pathway. Our results show for the first time in animal model that deficient synthesis of GD3 and consequently other b‐series gangliosides, beside effects on membrane lipid homeostasis, is also probably associated to alterations in complex signaling pathways. OP3 CHARACTERISTICS OF GLIAL CELLS OF THE SUBPLATE ZONE IN DIFFUSE AND FOCAL PERIVENTRICULAR WHITE MATTER INJURY Pogledić I1,2, Fallet‐Bianco C2, Gressens P2, Kostović I1, Verney C2 1Croatian Insitute for Brain Research, Medical School, Zagreb, Croatia, 2UMR 676 Inserm‐Univ Paris 7‐ Hopital Robert Debre, Paris, France Focal and diffuse periventricular white matter injuries (PVWMI) are the main histopathological brain lesions detected in preterm infants. Recent studies have shown that children born very preterm were more likely to develop neurological, cognitive and/or behavioral handicaps than children born preterm. Our study analysed different phenotypic expression of glial cells observed in diffuse and focal PVWMI on brain tissue (n=28) obtained from very preterm (24‐30 gestational weeks ‐ gw) and preterm (31‐35 gw) infants in frontal cortex. Antibodies used: for astrocytes (Glial Fibrillary Acidic


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Protein‐ GFAP; Monocarboxylate transporter ‐ MCT‐1; vimentine); microglia‐macrophages (Ionized Calcium Binding Adapter Molecule 1 ‐ Iba‐1); vessels (MCT1 and CD34) and pre‐oligodendrocytes (Olygodendrocyte Transcription Factor 2 ‐ Olig2 and Myelin Basic Protein ‐ MBP). On hematoxylin‐ eosin staining the different types of PVWMI were identified.The phenotypic features and morphology of glial cells in cortex (layer V/VI), superficial subplate, deep subplate and white matter below the subplate were described. Focal lesions displayed higer density of GFAP+ astrocytes in late preterms. These cells show more developed cell processes in very preterm infants vs. late preterm. Iba1 had the highest expression in the white matter, indicating activation (macrophages). Microglia was less activated in deep and superficial SP. Diffuse lesions showed actrocytes with vacuoled appearence in layers below the cortical plate in late preterms. The expression of MCT 1 in astrocytes was not different in PVWMI compared to controls, except in superficial and deep SP where it showed strong expression in vessel walls in PVWMI. This study identifies criticial differences in glial phenotypic expressions in diffuse compared to focal lesions in PVWMI. Supported by Croatian Ministry of Science grant No. 108‐1081870‐1876 (IK) and scholarship from the French Government (IP). OP4 GENDER AND AGE RELATED DIFFERENCES IN RECOGNITION OF FACIAL EXPRESSIONS OF EMOTIONS Barbir Lovorka1, Gregorić Boris1, Harhaj Ana1, Ilakovac Vesna2, Heffer‐Lauc Marija1 1Department of Medical Biology, University of Osijek School of Medicine, J. Huttlera 4, Osijek, 2Department of Biophysics, Medical Statistics and Medical Informatics, University of Osijek School of Medicine, J. Huttlera 4, Osijek The seven basic emotions: anger, sadness, fear, surprise, joy, contempt and disgust are cross‐culturally readily recognized. Other complex emotions are composed out of basic seven. Although voluntary muscles allow certain degree of deception, autonomous component of facial expression provides insight in real intentions. We assumed that recognition of facial expressions of emotions is the innate ability of individual brain with gender specific pattern. Survey constructed out of 13 photos with basic and complex emotions expressed by professional male and female actors was given to 217 participants (147 female/ 70 male) of different age. We tested the recognition of facial expressions of following emotions: joy, sadness, fear, contempt, disgust, surprise, jealousy, despair, confusion, shame, worry, anxiety and anger. Our results show that female participants were better in recognizing emotions on male and female faces. Ali participants were better in recognizing facial expressions in female faces than in male faces. Ali participants showed remarkable precision in detecting joy (100% on male face/97.7% on female face) and surprise (77.4% on male face/ 87.1% on female face). They also had a low ability of recognizing sadness (22.6%) and despair (22.6% ) on male faces and recognizing worry (22.6%) and anger (28.1%) on female faces. Particularly, male participants had low ability in recognizing sadness on male (5.5%) and female faces (6.9%), despair on male faces (5.5%) and worry on female faces (7.8%). Female participants had low ability in recognizing anger on female faces (18.4%), but moderate ability in recognizing anger on male face (49.3%). On the other hand, female participants were better in detecting contempt and shame on female faces (32.3%, 44.7%, respectively), comparing to contempt and shame in male faces (19.4%, 17.5%, respectively). Male participants were also better in detecting shame on female faces (21.2%), than in male faces (6.5%). Gender specific pattern in recognizing emotions could explain some misconceptions and traditional roles humans played during cultural evolution. OP5 STIMULATION OF FUNCTIONAL VISION IN CHILDREN WITH PERINATAL BRAIN DAMAGE Alimović S1, Katušić A1, Mejaški‐Bošnjak V, MD, PhD2

1Little house, day care center for rehabilitation of children with visual impairment and multiple disabilities, Baštijanova ID, 10000 Zagreb, 2Children's Hospital Zagreb, University of Zagreb, Medical School Cerebral visual impairment (CVI) is one of the most common causes of bilateral visual loss, which frequently occurs due to perinatal brain injury. It may happen that CVI stays unnoticed even through


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life, although child can have consequent delay. In early age normal visual function has a remarkable influence on the overall development of the child. It has great impact on acquisition of basic comprehensions which are fundamentals of further development. Therefore, the importance of early detection of visual problems and early intervention is very important. The aim of present study is to determine specific visual functioning of children with perinatal brain damage. It is also our aim to investigate the influence of visual stimulation on development of functional vision at early age of life. We initially assessed 30 children with perinatal brain damage (mainly with intraventricular hemorrhage and periventricular leukomalacia) up to 3 years of age, who were referred to day care center for pediatric low vision "little house" from neuropediatritions, ophthalmologists and other. Visual stimulations were carried out with children who had any visual impairment. Visual stimulations were determined according to functional vision assessment, individually for every child. Some of the children were stimulated with light stimulus, some with different materials under the UV light, and some with bright color and high contrast materials. Children were also involved in program of early stimulation of overall sensory motor development. Goals and methods of therapy were also determined individually, based on observation of child's possibilities and needs. After one year of program, reassessment was done. Results for visual functions and functional vision were compared to evaluate the improvement of the vision development. The results have shown that there was significant improvement in functional vision, especially in visual attention and visual communication. OP6 EFFECTS OF VIBROTACTILE STIMULATION ON THE CONTROL OF SPASTICITY AND MOVEMENTS FACILITATION IN CHILDREN WITH CEREBRAL INJURY Katušić A1 , Alimović S1 , Mejaški‐Bošnjak V2 1Day care center for rehabilitation ‐ Little house Zagreb, 2Children's Hospital Zagreb, University of Zagreb, School of Medicine. Afferent signals from the muscle's proprioceptors play important role in the control of spasticity and in the facilitation of movements. Peripheral afferent pathway enables the restoration of connections with supraspinal structures and so they includes mechanism of synaptic inhibition in the performance of normal movement. Different sensory stimuli, as vibrotactile stimulation, excite muscle's proprioceptors which than send sensorimotor information via spinal cord. In this way afferent signals promote cortical control and modulation of movements. The goal of this study is to evaluate the effects of vibrotactile stimulation on the spasticity and motor performance in children with cerebral injury. Subjects included in this study were 13 children who were developing the classification of spastic cerebral palsy. For all children perinatal brain damage was documented by medical reports and neonatal brain ultrasound scan. At the mean age of 3 years and 6 months subject underwent the assessment of motor development by GMFM‐88 test (Gross Motor Function Measurement). Gross Motor Classification System (GMFCS) has been used to classify functions of lower extremities. Therapeutic intervention was conducted once a week during 2 months. All subjects were stimulated with vibrotactile stimuli in duration of 20 minutes in order to reduce spasticity. After the ending of the treatment subjects underwent second assessment of motor performance and the classification of lower extremities functions. The results have shown that there was a significant improvement in motor performance, what has been seen in the facilitation of rotations, better postural trunk stability and head control and in greater selectivity of movements. Further randomized control trial investigations with bigger sample are needed to gain deeper insight in the role of vibrotactile stimulation in the facilitation of normal movements.


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POSTER WALK II

P6 COGNITIVE NEUROSCIENCE PP52 EARLY PHONOLOGICAL DEVELOPMENT AT CHILDREN WITH EARLY BRAIN INJURY Blaži D, Kovač‐Gornji N University of Zagreb, Faculty of Education and Rehabilitation Sciences There are still not enough reliable information on influence of early brain injury on children's communication and language development. Main reason lies in the fact that the population of children with brain damage is very hetergoeneous considering the stage at diagnosis, category, lesion location and other factors. However, researches conducted so far indicate delay in early language acquisition on children with early brain damages in comparison with children without brain injury. They also indicate some difficulties in language tasks of greater demand. The main objective of this work is to determine the difference between early phonological development on children with early brain damages and their peer groups without brain damages and to discover the nature of that difference. We have also tried to define the connection between different stages and categories of early brain damages and the development stage of phonological abilities. Our sample included 9 children with early brain damages (introventricular hemorrhage, hypoxic‐ishemic brain damage, agenesis of the corpus callosum ). Their chronological age ranked from 8 months to 2,06 years. Test group included children without brain damages which were the same age and sex as children in the first group. The research variables concern prelingual and early lingual development. We have also analysed vocalization and the phenomena of canonical babbling and the first word. The results indicate the delay in manifestation of these variables on children with early brain damage in comparison with children from the control group. PP53 GAZE DIFFERENCES IN PROCESSING PLEASANT AND UNPLEASANT COMPLEMENTARY PICTURES Budimir Sanja1, Palmović Marijan2

1University of Zagreb, Croatian studies, Psychology department, 2University of Zagreb, Laboratory for psycholinguistic research The International Affective Picture System (IAPS) is a set of standardized emotionally evocative color photographs developed by NIMH Center for Emotion and Attention at the University of Florida. It contains more then 900 emotional pictures indexed by emotional valence, arousal and dominance. However, when the IAPS pictures were used in studying emotions with the event‐related potentials, the results have shown a great deal of variation and inconsistency. In this research we analyzed pleasant and unpleasant pictures with an eye‐tracker in order to determine to what the participants turn their gaze. Considering that perception can be influenced by color, edges, luminosity and contrast and since all those factors are collapsed on the pictures in IAPS, we compared color pictures with same black and white pictures. In previous eye‐tracking IAPS research we analyzed 12 emotional pictures and showed a significant difference in the way participants look at the pleasant and unpleasant pictures. Now, we wanted to see is that difference due to some features of pictures or just emotional factor on the picture. All pictures are divided into figure and background, and based on this division, 8 complementary pictures were selected differing by the content on the figure area (pleasant, unpleasant). In the present study three problems related to the IAPS pictures were addressed: ‐ Differences among participants in looking at the figure and background area of emotional picture ‐ Differences on gaze parameters depending on emotional valence of the picture (pleasant, unpleasant)


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‐ Differences on gaze parameters depending on the color of the picture (color / black‐white) Participants in the eye‐tracking experiment were 25 psychology students with normal vision. Every participant saw all 8 presented pictures in color and same pictures in black/white version (4 pleasant, 4 unpleasant). This makes 200 analyzed units for color pictures and 200 analyzed units for black and white pictures. Every picture is divided into figure and background. There is a significant difference in number of fixations, average duration and dwell between background and figure, score is higher for the figure part for most of the analyzed pictures while no difference was found in number of entrances between figure and background. Both, pleasant and unpleasant pictures have higher average duration and dwell for figure then for the background while pleasant pictures have also higher number of fixations in the figure then in the background area. Also, both, colored and black‐white pictures have higher average duration and dwell for the figure area while black‐white pictures also have higher number of fixations in the figure area. Participant spend more time looking at the figure part of the picture then background but there is no difference in the gaze depending on emotional valence and color of the complementary pleasant/unpleasant pictures. PP54 EARLY COMMUNICATION DEVELOPMENT OF SOCIALLY DEPRIVED CHILDREN: ARE THERE SIGNS OF "INSTITUTIONAL AUTISM"? Cepanec Maja, Gmajnić Iva, Ljubešić Marta Department of Speech and Language Pathology, Faculty of Education and Rehabilitation Sciences, University of Zagreb Social deprivation‐induced changes in brain structure and function have been well described in both non‐human primates and other experimental mammals. However, there are few data on effects of early negative influences on the structure and/or functions of human infant`s brain. In the study of Romanian orphans, Chugani et al (2001) found decreased metabolic rate in the prefrontal (orbitofrontal and infralimbic) cortex, amygdala, hippocampus and the lateral temporal cortex. The affected brain structures were quite similar to those putatively involved in pathogenesis of autism. It seems that social deprivation in infants, as in other mammals, can result in autism‐like symptoms. Moreover, it is known for more than 40 years that institutionalized and autistic children display similar patterns of behavior. The aim of this study was to analyze early communication development in institutionalized children (N=24; age‐range 12 to 24 months), and to compare it with typically developing and age‐matched peers raised in family settings (N=24). Paired samples method was used. Our findings indicate that 75% of institutionalized children displayed conspiciously low performance and differed from their peers in all three sets of variables – social (p=0,004), speech (p=0,000), and symbolic (p=0,000). As a group, they display some autism‐like symptoms (non‐systematic response to their own name, age‐inappropriate joint attention skills, delay of appearance of first word(s), delay in language comprehension and babbling, decreased interest to play with different objects, lack of functional and symbolic play). However, institutionalized children differ from children with autism in two important diagnostic criteria – they show interest for social environment and communicate for noninstrumental purposes. In conclusion, institutionalized children show delayed pattern of early (socio)cognitive, communication and language development, but they do not show a full range of symptoms typically appearing in children with autism. These findings need to be replicated with a larger sample, because some other studies indicated that the incidence of autism may be increased in institutionalized children. PP55 COMPARISON OF EMOTION RECOGNITION IN FACIAL EXPRESSION AND MUSIC Gašpar Tina, Jurić Iva, Labor Marina, Dumančić Dijana, Ilakovac Vesna, Heffer‐Lauc Marija School of Medicine, J.J.Strossmayer University of Osijek Introduction. Expressions of basic emotions are characteristic and universal for all races. All people can express them on face, using conscious and unconscious muscles, and recognize them at other


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people's faces. Lower left temporal cortex, typically at the right side, memorizes and recognizes characteristic lineaments. It is not clearly stated how good is the usual ability to recognize emotions because of their shortness (micro expressions), and the recognition doesn't have to be conscious willing process. We assumed that visual recognition of facial expressions is selected over recognition of emotions via auditory stimuli so we compared success in recognizing emotions in human faces and in classical music works. We also wanted to know if there is a difference in recognizing emotions between examinees at early and late puberty ages. Subjects and Methods. We surveyed 90 students attending the 5th grade of elementary school and 87 students attending the 3rd grade of high school from Osijek (Croatia), 177 students in total. The questionnaire consisted of three parts: the first part included basic information about examinees, and in the second and third part students had to link 8 photographs of different facial expressions and 8 pieces of classical music work with 8 associated emotions (worry, happiness, sadness, surprise, nerve, anger, fear and jealousy). Each correct answer was given one point. Results. Median number of correct answers in recognizing emotions from facial expressions was 5 (95% CI 5 to 6). Emotion recognition based on classical music pieces was significantly less successful (median number of correct answers was 2; 95% CI 2 to 3; Wilcoxons' P<0.001). High school students did better in recognizing emotions from facial expressions then elementary school students (Mann‐Whitney test, P=0.002). Girls achieved better score then boys in recognizing emotions from both facial expression (Mann‐Whitney test, P=0.001) and musical pieces (Mann‐Whitney test, P=0.020). Success in recognizing emotions from musical pieces was associated with higher math grades (Jonckheere‐Terpstra test, P=0.018). Conclusions. Emotions are far better recognized if presented on human faces then in music, possibly because emotion recognition based on face expressions is one of the oldest communicational skills in human society. Female dominance in emotion recognition could be respond to necessity of communication with the newborns during nursing time. Mastery of recognizing emotional content of music and mathematical skills both require ability of multivariable function handling and insight in abstract structures which might be the ground of the association of mathematics and musically expressed emotions. PP56 COGNITIVE ABILITIES AND LANGUAGE COMPREHENSION IN PRESCHOOL CHILDREN WITH PERINATAL BRAIN LESION Ivšac Pavliša J, Šimleša S, Ljubešić M Developmental Neurolinguistic Lab, Faculty of Education and Rehabilitation Sciences, University of Zagreb Perinatal brain lesion is a risk factor for development, making the parents of such children particularly worried about consequences it may have on the child's cognitive and language development. Although the literature findings on the outcome of perinatal brain lesion are inconsistent, most of the studies found a positive general outcome, but also subtle deficits that affect children's academic success. Since language comprehension and cognitive abilities influence learning abilities at school, we wanted to know how six‐year olds who were selected based on pathological ultrasonographical findings (ischemic or hemorrhagic brain injury) would perform on subtests of Wechsler battery WISC and Reynell Developmental Language Scale, compared with controls. The second issue we investigated was whether in children suffered from perinatal brain injury cognitive abilities predicted the level of language comprehension in the same way as in children without perinatal brain lesion. The relation between cognitive and linguistic abilities is still a controversial one, and different relation would mean that these two groups of children have different structure of abilities probably connected with perinatal brain lesion. We examined 40 children suffered from perinatal brain injury and 40 age‐matched children without perinatal risk factors. Our results showed that the groups differed more in linguistic than in cognitive variables. Also, the two groups showed different relation patterns between cognitive abilities and language comprehension. Cognitive abilities were statistically significantly associated with language comprehension in children suffered from perinatal brain injury, while control children did not show statistically significant difference


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between cognitive and linguistic parameters. Since a part of participants with perinatal brain lesion had language difficulties, we presume that they rely on cognitive abilities in order to overcome and compensate for language shortcomings. PP57 SPATIAL AND TEMPORAL MEASUREMENTS OF EYE MOVEMENT IN CHILDREN WITH DYSLEXIA: INDIVIDUAL DIFFERENCES Kuvač Kraljević Jelena and Palmović Marijan Laboratory for Psycholinguistic Research, Department of Speech and Language Pathology University of Zagreb Eye‐tracking is a relatively new method in reading studies. The method allows for researching two crucial levels underlying reading: visual and cognitive. Reading research using eye‐tracking brought about three main approaches to reading: minimal‐control view, visual/oculomotor control view and recent, cognitive‐control view. These approaches vary in their understanding of the relationship between eye movement and linguistic processing of the text: ‐ in minimal‐control view there is no relationship; ‐ in visual/oculomotor control view eye movement is only indirectly relates to linguistic processing; ‐ in cognitive‐control view linguistic processing of the text determines the eye movement. The aim of this paper is to show the differences in eye movements of three children with dyslexia using the principles of cognitive‐control view. The collateral aim is to develop methodology to use eye‐tracking technique to dissociate between various subgroups of dyslexia. Namely, despite well known definitions and vast literature on dyslexia, manifestations of dyslexia on the individual level are highly varied. All three children were tested behaviorally using set of language tests for language behavior assessment. Two children had low scores on most language tests, and all three children had poor level of reading and writing. Using SMF s iView X Hi‐Speed eye‐tracking system all children had to read a text silently while the device recorded the eye movements, i.e. fixations (number, position, duration) and saccades (number, speed, magnitude). Spatial and temporal features of reading were obtained using these data: perceptual span, gaze durations, total viewing time, etc. Although all three children have the same diagnose ‐ dyslexia, analysis of their eye movements during reading shows different saccade‐fixation patterns. The differences were obtained on a number of variables: number and length of fixations and number of regressive (or back) saccades. The results are compared with the results obtained in the group of typically developing children who show a clear pattern of efficient reading. PP58 EFFECTS OF PRESENTATION TIME AND INTERPOLATED ACTIVITY ON FALSE MEMORY Manenica Ilija, Prenđa Suzana University of Zadar False memory refers to information about some past event, which is substantionaly different from that event. The phenomenon of false memory is predominantly based on associations between memorised elements concerning the event, and some other elements which are not related to the event itself. There are two models of explanation of false memory phenomenon. The model of spreading activity throughout semantic network (SASN), holds that mental elements, such as words or images, are stored in memory together with their associations. That means, that a list of mutually associated words activates semantic network of other words which are not on the list, but associated with them. The stronger the associative links are, the higher probability of remembering the word out of the list (critical word). The other model of memory control of presented material (MCPW) includes mnemonics as the way of memorising, where critical words may indirectly appear as associative elements. The aim of this study was, by the use of Deese‐Roediger‐McDermott paradigm (DRMP), to check the two models. DRMP consists of 10 lists with 15 words each, which are visually presented to subjects. Words on the lists have associative connections with some other words, i.e. critical words. The probability of choosing the critical word as presented was almost the same as the probability of remembering any presented word. The experimental situations differed according to


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presentation times and interpolated activity. In one experimental situation, words from the lists were presented visually one after the other, in time intervals of 100, 300, 500 and 3000 ms. The subject's task was to verbally reproduce, as many words as possible from the list. In the other situation the procedure was the same, but during the list presentation, the subject had to count backwards by three from a given three digit number. Analysis of results showed an increase in false and real memory as the time intervals increased from 100 to 500 ms, but no effects of interpolated activity. At the presentation interval of 3000 ms, however, there was a significant fall in false and an increase in real memory. Contrary to this, in the situation with interpolated activity, false memory was significantly higher and real memory fell down to the presentation level 100 ms. This suggests that subjects were repeating, and consequently remembering better, presented words during 3000 ms interval without interpolated activity. These results support the hypothesis of the SASN model being better for explanations of false memory results at shorter presentation intervals, and the MCPW at longer. PP59 APPLICATION OF THE CONTROLLED ORAL WORD ASSOCIATION TEST IN A CROATIAN SAMPLE Mimica Ninoslav1,2, Žakić Milas Danijela1, Joka Svetislav1, Kalinić Dubravka1,2, Folnegović Šmalc Vera1,2, Harrison John E3,4 1University Department of Psychiatry, Psychiatric Hospital Vrapče, Zagreb, Croatia; 2School of Medicine, University of Zagreb, Zagreb, Croatia; 3CogState Ltd., Warminster, United Kingdom; 4Imperial College, Division of Neurosciences & Mental Health, London, United Kingdom. Introduction: The Controlled Oral Word Association Test (COWAT) is widely used in clinical neuropsychology, primarily as a measure of verbal fluency, but also as a measure of executive skills such as working memory, planning and strategy. Verbal fluency tasks such as the COWAT appear to be relatively easy task to perform and yet highly selective lesions have been observed to cause fairly substantial deficits in performance. In order to perform this task successfully, the study participant must develop a strategy for retrieving words beginning with the stimulus letter, whilst complying with rules that forbid them to mention proper nouns and serial word stems with changed word endings. Study participants are also requested not to repeat words. The need to recall what has previously been said presumably represents a burden to working memory. The COWAT forms part of the Neuropsychological Test Battery which has been profitably employed in a number of clinical drug trials, including studies of AN1792‐201 (an amyloid lowering treatment for use in patients with Alzheimer's disease) and a recent study of the chelating agent PBT2. The COWAT was also employed in the EXPRESS trial of rivastigmine in patients with Parkinson's disease dementia and routinely features in cognitive assessment batteries for a variety of both psychiatric and neurological disorders. Although the COWAT is increasingly employed in international trials, very little is known about the utility of the English version of the test in non‐English speaking populations. English versions of the test employ letter stimuli for which native English speakers are most fluent, and these letters may be inappropriate for use with Croatian speakers. Aim:The aim of this study was to find out the word frequency for all letters in the Croatian alphabet in order to identify the most appropriate letters for use with Croatian speaking study participants. This methodology replicates the investigation originally conducted with English speakers and by which the most appropriate letters were selected for use. We also sought to determine whether there are any differences between Croatian and English speakers with respect to fluency. Subjects and methods:There were 77 healthy participants, age ranged from 18 to 52 years. Among them, there were 21 (27.3%) male and 56 (72.7%) female. All study participants were given 1 minute per letter for all the letters of the Croatian alphabet. Results: In our sample we detected that participants generated most frequently words starting with the letters "K" (M‐13.32) "P" (M‐12.01), "S" (M‐11.77) and "M" (M‐11.48). Conclusion: Our preliminary results suggest that there are differences among most frequent letters in COWAT between Croatian and English speakers. We recommend the use of the letters "K", "P", "S" and "M" in studies of verbal fluency (COWAT) performance conducted with Croatian speaking individuals.


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PP60 KOGNITIVNI SLUŠNI EVOCIRANI POTENCIJALI U DJECE S GOVORNO‐JEZIČNIM POTEŠKOĆAMA Munivrana Boška, Išgum Velimir, Orlović Joško, Marn Borut Poliklinika SUVAG, Zagreb, Hrvatska; KBC Rebro, Zagreb, Hrvatska, FER, Zagreb, Hrvatska, Klinika za dječje bolesti, Zagreb, Hrvatska U ovom istraživanju ispitivana je sposobnost percepcije i diskriminacije zvučnog i govornog podražaja. Provedena su dva eksperimenta.U prvom su kao podražaj korištena dva čista tona (1kHz i 2kHz), a u drugom dva dvostruka sloga, sastavljena od jednog suglasnika i jednog samoglasnika hrvatskog jezika odabranog prema karakteristikama tzv. optimalnog filtera (frekvencijskog područja u kojem se određeni glas najbolje percipira kao glas hrvatskog jezika). Ispitanici su bili 18‐ero djece, s govorno jezičnim poteškoćama, u dobi od 7 do 9 godina, te kontrolna skupina djece, iste dobi, s urednim govorno jezičnim razvojem. Podražaji su bili prezentirani u odd ball paradigmi, te se od ispitanika tražila svjesna reakcija. Mjerene su latencije i amplitude analiziranih (P1,N1,P2,N2 i P3) valova, te vrijeme reakcije i broj točnih odgovora. Ispitivanje je pokazalo da postoje bitne razlike u morfologiji dobivenih valova (amplitude i latencije) djece s govorno‐jezičnim poteškoćama u odnosu na kontrolnu skupinu djece. Ključne riječi: ERP, govorno jezične poteškoće, djeca PP61 POSTADOLESCENT CIRCUITRY REORGANIZATION OF ASSOCIATIVE LAYER IIIC PYRAMIDAL NEURONS IN THE HUMAN PREFRONTAL CORTEX Petanjek Zdravko, Zeba Martina, Uylings BM Harry, Kostović Ivica Department of Neuroscience, Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Šalata 12, 10000 Zagreb, Croatia That human cognitive and emotional development continues through adolescence is a general fact accepted among developmental psychologists and in past decade supported by extensive data of functional brain development. These findings have been taken as indicators of protracted synaptic pruning or/and protracted dendritic growth. However, as per our knowledge, there are no available data of cortical circuitry development that can support this assumption. The aim of this study was to examine adolescent and post‐adolescent dendritic growth and dendritic spine density on key elements of circuitry involved in processing of higher cognitive functions, the layer IIIC pyramidal neurons in dorsolateral prefrontal cortex (Brodmann's area 9). We used available quantitative data of dendritic morphology and dendritic spine density obtained from 25 specimen of Zagreb neuroembryological collection. Data obtained per subject were used as a row data for statistical analysis between following groups: childhood (2‐8 years), adolescence (9‐18 years), early adulthood (19‐30 years), adult (31‐65 years) and aging (66 years+). Except intermediate segments, length and branching pattern of dendrites has not changed significantly. However, we found massive statistically significant decrease in dendritic spine density that occurred during adolescence, but also continued through early adulthood for all segment type analyzed. In parallel, the length of intermediate segments decreased for about 20%. Some differences in pattern of dendritic spine density between different segment types were seen. Dendritic spine density in adulthood was highest in the most distal segments of apical oblique dendrites, which showed also largest and most protracted decrease in spine density compared to proximal oblique and basal dendrites. Distal dendrites are known to be predominantly innervated by intracortical and associative cortico‐cortical fibers. The maturation of layer IIIC pyramidal neurons might represent a biological basis for protracted cognitive development and, as such, may be highly influenced by education and social environment. Besides biomedical, educational and social implications, these results also point to highly protracted environmentally driven plasticity in associative cortex that influence circuitry development, suggesting mechanisms how and for how long the environment reshapes synaptic circuitries that builds biological basis of individuality.


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PP62 EFFICACY IN FITT'S TAPPING TASKS ACROSS MENSTRUAL CYCLE Šimić Nataša, Lovrić Marijana, Šuto Ana University of Zadar, Faculty of Phylosophy Mostar, Primary School "Dr. Jure Turić" Gospić Numerous findings suggest that menstrual cycle related variations in sex hormones influence cognitive processes (Kimura, 2002), psychomotor abilities (Mead and Hampson, 1997) and mood (Campton and Levine, 1997). Although different studies in this area have not came up with the same results, high levels of progesterone in luteal phase have been suggested to reduce functional cerebral asymmetry (Hausmann et al., 2008).Given that cerebral asymmetry indeed changes across menstrual cycle, it seems likely that differences in performance of Fitt's tapping tasks (FTT's) with right and left hand would be less pronounced in luteal phase with regard to phases of lower sex hormones level. Additionally, in phases of higher sex hormones levels better efficacy can be expected.Thus, the aim of this study was to examine changes of efficacy in performance of FTT's across menstrual cycle.Twenty‐four normally cycling women with a mean age of 21.7 years participated in this study. They were tested four times, during menstrual, late follicular, luteal and premenstrual phase. Subjects performed series of FTT's, difficulty range from one to four bits. The working time for every task was one minute. Twelve subjects started performing the tasks with right, and other twelve with left hand. All subjects used right as their dominant hand.Results suggest a decrease in performance of FTT's during luteal phase of a menstrual cycle, with tendency of decrement in function of task difficulty. Additionally, results indicate an increase of efficacy in menstrual phase, when levels of estradiol and progesterone were low. Furthermore, the results did not show a decrease cerebral asymmetry during luteal phase, as expected.Generally speaking, these results suggest that efficacy in performance of FTT's shows similar pattern of changes as in tasks which males usually solve better than females (i.e. spatial tasks). PP63 EFFICIENCY IN SPATIAL AND VERBAL TASKS DURING THE MENSTRUAL CYCLE Šimić Nataša, Santini Maria Department of Psychology University of Zadar Numerous investigations have shown that changes in levels of sex hormones are responsible for changes in mood and efficiency during the menstrual cycle. It has been reported that high levels of estrogen and/or progesterone improved the skills such as verbal fluency and articulation for which females typically show better results than males. In contrast, low levels of these hormones improved the spatial abilities in which male usually show a better results (Kimura, 1996; Hampson, 1990; Hausmann et al., 2000). Using various verbal and spatial tasks at different levels of complexity, the aim of the study was to explore the effects of the menstrual cycle on cognitive functions, as well as the relationship between efficiency and masculinity/femininity, as personality characteristic, during different phases of menstrual cycle. Seventeen female subjects (18 to 21 years), with a regular menstrual cycle of 28 days and not using oral contraceptives, took part in this study. Subjects performed the verbal fluency and mental rotations tasks during the early and late follicular phase, as well as midluteal phase. Endler's State Anxiety Questionnaire was administered before performing the tasks in each phase. For assesssments of masculinity and femininity, the subjects also filled the Bern Sex Roles Inventory. The results showed the best mental rotation task performance in the early follicular phase, which is characterised by a low level of estrogen and progesterone. The most pronounced during this phase of menstrual cycle was also the masculinity. In contrast, the best performance in tasks of verbal fluency occurred during the midluteal phase, when levels of estrogen and progesterone are the highest. Femininity and cognitive component of anxiety did not change across the menstrual cycle. The results of this study showed that only changes in efficiency in spatial tasks were in relation with changes masculinity.


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PP64 USEFULNESS OF DRAWING AND WRITING TEST WITH BOTH HANDS IN SUBJECTIVE AND OBJECTIVE DETERMINATION OF DOMINANT HEMISPHERE Šnajder Darija, Labak Irena, Kostović‐Srzentić Mirna, Benčić Mirta, Ništ Marina, Ilakovac Vesna, Heffer‐Lauc Marija Medical Faculty Osijek, School of Nursing Zagreb, Department of Mathematics Osijek, Department of Biology Osijek Brain lateralization is common term used to describe dominance of one brain hemisphere over another for specific function. Right hand dominance in writing is in correlation with development of communicative gesticulation and afterwards with the development of speech in same hemisphere. Nonetheless, part of cognitive functions included in drawing function might have origin in opponent hemisphere. We were interested to find out weather the performance of function common to be processed in one hemisphere would be more precise than the performance of the function processed by both hemispheres while using non dominant hand. We assumed that some people perform both functions with same accuracy and that they are not aware of their own capability in using the other hand. Participants were asked to write two words and draw a particular object ‐ vase with flowers. The sample comprised 1546 children from 19 primary and 8 high schools in Osijek area, from which 1030 primary school children from 1st till 4th grade and 516 high school children from 3rd and 4th grade. 815 girls and 731 boys were included in the research. A leaflet consisting of two parts was used in the research. The first part was a questionnaire consisted of 10 questions for subjective analysis of commonness of using right hand or leg. The second part consisted of a drawing and writing test done with both hands. After we validated the drawing and writing test, children were put into three groups: right ‐ handers, left ‐ handers and ambidextrous. The frequency of answers on commonness of using right hand/leg and our categorization of children based on hand dominance were tested with Chi‐square test. In our sample are 1384 (89,5%) right‐handers, 120 (7,8%) left‐handers and 42 (2 ,7%) ambidextrous. Among primary school children there are 915 (88,8%) right‐handers, 77 (7,5%) left‐handers and 38 (3,7%) ambidextrous. Among high school children there are 469 (90,9%) right‐handers, 43 (8,3%) left‐handers and 4 (0,8%) ambidextrous. Subjective understanding of hand preference is the same as shown in security test for same hand in all questions but those about sport activities such as ball throwing, hitting the ball with the foot, following the rhythm of the music by foot, which are considered to be done automatically. Subjective understanding compared to drawing and writing test deviated in case of ambidextrous who consider themselves right‐handers. The fact that the number of ambidextrous is greater in primary schools than in high schools brings us to conclusion that training of the right hand decreases the ability of using both hands equally for both tested functions. PP65 EYSENCK’S TEMPERAMENT TYPOLOGY AND EVOKED POTENTIALS Tatalović Vorkapić Sanja, Tadinac Meri Department of Preschool Education, Teacher Education College, Rijeka The aim of this study was to examine the relationship between latencies and amplitudes of evoked potentials (N1, P2, N2, P3 & Sw) and four temperament types defined on the basis of Eysenck's E and N dimensions: phlegmatic (E‐N‐), choleric (E+N+), melancholic (E‐N+) and sanguine (E+N‐). According to Pavlov’s and Eysenck's arousal theories, and studies of neurological differences between the temperament types (Robinson, 1982, 2000, 2001), it was expected that extraverts and sanguines would be characterized by a lower natural frequency (predominance of inhibition), while introverts and melancholics would show a higher natural frequency (predominance of excitation). Theoretically, the excitation dominance should be positively correlated with lower EP‐amplitudes, but when the complex oddball tasks, reducing the habituation effect to the minimum, were used in the EP‐studies, higher EP‐amplitudes were determined in extraverts. Despite these inconsistent findings, the lowest EP‐differences were expected between emotionally stable subjects: sanguines and phlegmatics, and


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the highest between melancholics and cholerics. Furthermore, phlegmatics should have the longest EP‐latencies, while cholerics were expected to show the highest EP‐amplitudes. A sample consisted of N=54 female psychology students, within the age range 19‐23 years, all right‐handers and without previous psychiatric/neurological illnes. Evoked potentials were measured by using the standard visual oddball paradigm in two trials within the time interval of 2‐3 minutes. Since the marking of the evoked potentials was manual, EP‐latencies were used as a marking criterion, and their values are therefore the same in the first and second trial, while the amplitudes differ. The participants were divided in four groups according to the median on Extraversion and Neuroticism subscales: phlegmatic (E‐N‐)=( E≤17 & N<8), choleric (E+N+)=( E>17 & N≥8), melancholic (E‐N+)=( E≤17 & N≥8) and sanguine (E+N‐)=( E>17 & N<8). The analysis of EP amplitudes mostly showed the expected pattern: the lowest amplitudes were found in phlegmatics and the highest in cholerics, with the sanguines and melancholics in between. The longest N2‐latencies were determined in phlegmatics, and the longest P2‐ and Sw‐latencies in melancholics, which could be expected considering their low position on the E dimension. The electrophysiological differences among four temperament types were clearly demonstrated, with the patterns consistent with Pavlov’s and Eysenck’s arousal theories. Key words: Temperament types, evoked potentials, individual differences, students

P7 BASIC NEUROSCIENCE

PP66 EXPRESSION OF SEROTONIN TRANSPORTER IN PERIPHERAL BLOOD LYMPHOCYTES FROM PATIENTS WITH ALCOHOL DEPENDENCE Antica Mariastefania1, Matošić Ana2, Marušić Srđan2, Jernej Branimir1, Čičin‐Šain Lipa1 1Department of Molecular Biology, Rudjer Boskovic Institute, 2Department of Psychiatry, Clinical hospital "Sestre milosrdnice", Zagreb, Croatia Introduction: Serotonin (5HT) has long been implicated in diverse aspects of immune regulation, acting through both central and peripheral mechanisms. The cells of the immune system are capable to actively take up this amine from the surrounding plasma via serotonin transporter (5HTT, SERT) on the cell membrane. Serotonin transporter, a 12‐transmembrane domain protein, represents the main regulator of the magnitude and duration of 5HT signaling, and dysregulation of its activity and/or expression has been linked to various disorders. With regard to alcoholism, alterations of both systems, serotonergic and immune, have been shown in experimental and clinical studies. Increased alcohol intake is associated with decreased 5HT transmission and, on the other hand, with impaired immune cells function. Their mutual relationship is, however, poorly understood. The aim of the present work was to study the expression of 5HTT on peripheral blood lymphocytes from patients with alcohol disorder compared to control subjects. Methods: EDTA‐anticoagulated whole blood samples were obtained from healthy individuals and alcoholic patients at the admission to the hospital. Peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation. The presence of 5HTT was analyzed by use of flow cytometry. The mouse monoclonal antibody against 5HTT which can specifically bind to an extracellular domain of human 5HTT was used to label human PBMCs. Results: Almost all lymphocytes and most monocytes were positive for 5HTT. When examined by multicolor flow cytometry, 5HTTbright cells were among CD20+, CD3+, CD11+ and CD14+ cells. Results obtained on alcohol‐dependent patients will be presented. Conclusion: Data demonstrate expression of 5HTT by PBMCs, and indicate that flow cytometry could be a suitable method to test the in vivo immune regulatory effects of 5HT or drugs targeting 5HT transporter. Studies on lymphocyte 5HT system might contribute to understand the bidirectional interaction between the nervous and the immune system in alcoholism.


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PP67 EXPRESSION AND FUNCTION OF 5HT‐1A RECEPTOR IN BRAINS OF WISTAR‐ZAGREB 5HT RATS Bordukalo‐Nikšić Tatjana, Čičin‐Šain Lipa, Jernej Branimir Laboratory of Neurochemistry and Molecular Neurobiology, Rudjer Bošković Institute, Bijenička 54, HR‐10000, Zagreb, Croatia Introduction: By selective breeding for the extreme values of platelet serotonin level and platelet serotonin uptake, two sublines of rats with constitutionally upregulated/downregulated platelet serotonin transporter (5HTt) have been developed (Wistar‐Zagreb 5HT rats). In previous studies, these sublines demonstrated significant differences in the expression of platelet 5HTt at the level of both mRNA and protein. Study on 5HTt gene expression in brains did not show significant differences between the sublines, although neuropharmacological and behavioral studies indicated that changes in central 5HT homeostasis might be expected. Here, we searched for possible adaptive/compensatory changes in expression and/or function of cortical 5HT‐1A receptors of Wistar‐Zagreb 5HT rats, since 5HT‐1A receptors are, besides 5HTt, the most important regulators of serotonergic neurotransmission. Materials & Methods: Studies were performed on animals from the 15th breeding generation. Semi‐quantitative RT‐PCR was performed on RNA isolated from frontal cortex, using two housekeeping genes, »‐actin and cyclophylin B, as a reference. Functional analysis of cortical 5HT‐1A receptors was performed by radioligand binding method, using [3H]‐8‐hydroxy‐2‐(di‐N‐propylamino)tetraline (8‐OH‐DPAT) as the radioactive ligand. Results: There were no difference in mRNA expression of cortical 5HT‐1A receptors between the sublines of Wistar‐Zagreb 5HT rats. At the functional level, the sublines did not differ, either in number of cortical 5HT‐1A binding sites (Bmax) or in the receptor affinity for ligand (Kd). Conclusion: Constitutional alteration in serotonergic neurotransmission of Wistar‐Zagreb 5HT rat sublines, induced by up/downregulation of serotonin transporter, did not result in change in expression or function of cortical 5HT‐1A receptors in physiological conditions. Further studies will be aimed to the 5HT (5HT‐1 A, as well as the other) receptor expression and/or function after specific pharmacological challenge, when possible subtle differences between sublines could become more detectable. PP68 PROPOFOL ATTENUATES THE PHRENIC LONG TERM FACILITATION IN RATS SUBJECTED TO ACUTE INTERMITTENT HYPOXIA Carev M1, Valić M2, Pecotić R2, Karanović N1, Valić Z3, Pavlinac I2 and Đogaš Z2 1Department of Anesthesiology and Intensive Care, University Hospital Split, Split, Croatia, 2Department of Neuroscience, University of Split School of Medicine, Split, Croatia, 3Department of Physiology, University of Split School of Medicine, Split, Croatia Introduction: Long term facilitation (LTF) is sustained augmentation of respiratory motor output elicited by acute intermittent hypoxia (AIH), and is considered as a form of the central nervous system plasticity. It has been demonstrated in urethane‐anesthetized rats. Propofol is an intravenous anesthetic widely used in humans. So far, the influence of propofol anesthesia on LTF in the model of AIH has not been studied in rats. Our hypothesis is that, there will be a significant attenuation of the response under propofol anesthesia. Materials and methods: The phrenic nerve activity (PNA) was recorded in 14 adult, male, Sprague‐Dawley rats, bilaterally vagotomized, paralyzed and mechanically ventilated. The phrenic nerve recordings monitored central respiratory activity. Phrenic nerve was dissected using dorsal approach at the level of C5 nerve rootlet, mounted on bipolar silver wire electrodes. The rats were placed in a prone position in a stereotaxic instrument. The animals were divided in 2 groups; group U (urethane‐anesthetized, 1.2 g/kg i.p., n=7) and group P (propofol‐anesthetized, i.v. bolus 11.9 mg/kg + infusion 1.3 mg/kg/min, n=7). Rats were exposed to five, 3‐min hypoxic episodes (FiO2=0.09), separated by 3 minutes of hyperoxia (FiO2=0.5). Peak phrenic nerve amplitude (PNA), burst frequency (f), inspiratory time duration (Ti), expiratory time duration (Te) and total respiratory cycle duration (Ttot) were


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analyzed during the first hypoxia and 15, 30, and 60 minutes after the final hypoxic episode, and compared to the baseline values prior to the first hypoxic episode. Results: Isocapnia was successfully maintained throughout the protocol. There was a significant increase of peak phrenic nerve amplitude (66.7 ± 8.6%, p<0.05) in urethane‐anesthetized group 60 minute after the last hypoxic episode compared to the baseline values, i.e. the LTF was induced. There were no significant changes in PNA in the urethane‐anesthetized group at 15 and 30 minutes after the last hypoxic episode. In the propofol‐anesthetized group no significant changes of PNA were recorded at any time point after the last hypoxic episode, i.e. no LTF was observed. AIH did not elicit significant changes in the phrenic burst frequency, as well as in breathing rhythm parameters (Ti, Te, Ttot) in both groups at all 3 time points after the last hypoxic episode. Regarding the first episode of AIH, PNA was significantly increased in both groups compared to baseline values (172.3 ± 2.0% in the urethane‐anesthetized group vs 106.8 ± 16.9 % in the propofol‐anesthetized group, p<0.001), whereas phrenic burst frequency was significantly increased only in the urethane anesthetized group (53.0 ± 3.6 vs pre‐hypoxic 36.5 ± 3.7 bursts/min, p<0.01). PP69 RhoB PROTEIN IS INVOLVED IN CELL RESPONSE TO GENOTOXIC STRESS Cimbora‐Zovko Tamara1, Fritz Gerhard2 and Osmak Maja1 1Laboratory for Genotoxic Agents, Department of Molecular Biology, Rudjer Bošković, Zagreb, Croatia; 2Institute of Toxicology, University of Mainz, Mainz, Germany Cell exposure to genotoxic stress induces a network of signaling pathways in treated cells. Following a single dose of cisplatin to human laryngeal carcinoma HEp‐2 cells, as well as in cisplatin resistant CA3ST and CK2 sublines, we observed altered cell morphology and cytoskeleton organisation, suggesting the alterations in Rho GTPases activity. Therefore we screened the expression of representative Rho family members. Among several Rho GTPases analyzed, RhoB expression was downregulated in the cisplatin‐resistant sublines on both mRNA and protein levels, as determined by semiquantitative RT‐PCR and Western blot analysis, respectively. In addition, immunocytochemistry revealed that subcellular localization of RhoB was altered after cisplatin treatment in both HEp‐2 and CA3ST cells. Transient transfection of cisplatin‐resistant CA3ST cells with plasmid vectors expressing wild type, as well as farnesylated or geranylgeranylated RhoB fused to EGFP protein increased cisplatin‐induced cell death measured by flow cytometry, while silencing of RhoB expression with specific siRNA decreased sensitivity of parental HEp‐2 cells to cisplatin. Thus, apart from regulating cytoskeleton, Rho GTPases are also involved in other vital cellular functions, among others in cell response to stress. The involvement of RhoB protein in response to neuro cytotoxic compounds on neuronal/glial cell cultures is currently under the study, which could model situations in some neurodegenerative disorders. PP70 PRENATAL DEVELOPMENT OF NITRINERGIC NEURONS IN THE HUMAN BASAL FOREBRAIN Džaja D, Ažman D, Pletikos M, Judaš M Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Zagreb, Croatia We present the first evidence on prenatal development of nitrinergic neurons in the human basal forebrain (BF). We used Neurolucida equipment for qualitative and quantitative analysis of dendritic development of BF nitrinergic neurons selectively visualized by NADPH‐diaphorase histochemistry at three main periods of their prenatal development: (1) at 15 postconceptional weeks (PCW), when BF afferents temporarily reside in the «waiting» compartment of the subplate zone (SP); (2) at 22 and 26 PCW, when BF afferents relocate from SP to the cortical plate; and (3) in 13‐day‐old term newborn. We analyzed 202 neurons from 4 specimens of the Zagreb Neuroembryological Collection. At 15 PCW, nitrinergic BF neurons are already numerous but immature and mostly bipolar. They rapidly differentiate during the midfetal period, so that at 26 PCW several morphological types (bipolar, fusiform, multipolar, triangular, arciform) were clearly recognized. The same types were present in the newborn, when their dendritic domains seemed to attain adult sizes and shapes. In contrast to the adult human BF in which Ellison et al. (J Comp Neurol 260:233, 1987) described only


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small and medium size nitrinergic neurons, we found that fetal and newborn BF also contains quite large nitrinergic neurons which display morphology typical for magnocellular cholinergic BF neurons. Therefore, we suggest that large cholinergic BF neurons transiently produce NO during prenatal and perinatal period. Supported by Croatian Ministry of Science, Education & Sport grant No. 108‐10818701878 (to M.J.). PP71 SINGLE ELECTROCONVULSIVE SHOCK DOES NOT INFLUENCE COX‐2 AND HSP70 EXPRESSIONS IN THE RAT FRONTAL CORTEX AND HIPPOCAMPUS Hrelja A, Pilipović K, Župan G Department of Pharmacology, School of Medicine, University of Rijeka, Rijeka, Croatia Introduction: Electroconvulsive therapy is commonly used in psychiatry as a treatment for pharmacotherapy‐resistant cases of severe depression and other affective disorders but there are still many questions concerning the mechanisms underlying its therapeutic and side effects. The aim of our study was to investigate whether single electroconvulsive shock (ECS) affects expressions of COX‐2 inflammatory and heat shock HSP70 proteins in the rat frontal cortex and hippocampus. Materials & Methods: Experiments were performed on male Wistar rats (age 2‐3 months; weight 230280 g). Animals were randomly divided into five experimental groups. Rats of the control group were subjected to the sham ECS procedure. Other groups of animals received single ECS (100 mA, 50 Hz, 0.4 ms interval, 400 ms total duration) via earclip electrodes using an ECS apparatus (Ugo Basile, Italy). Animals were decapitated at different time points (0.5 h, 3 h, 12 h, 24 h) during the first 24 h after single sham ECS procedure or ECS‐induced seizures. Western blotting analyses of COX‐2 and HSP70 expressions were performed in the frontal cortex and hippocampus. Results: Cortical and hippocampal COX‐2 and HSP70 expressions in rats with ECS‐induces seizures were not significantly changed in comparison to the values registered in the sham‐ECS treated rats, at any of the time points examined. Conclusion: Our findings suggest that single ECS does not affect COX‐2 and HSP70 expressions in the rat frontal cortex and hippocampus, at different time points within the first 24 h after its application. PP72 HUMAN FOETAL SPERMATOGONIA EXPRESS NEURON‐SPECIFIC ENOLASE Ježek Davor1 Šklebar Duška2, Kozina Vivian1, Šklebar Ivan2 and Kos Marina3 1University of Zagreb, School of Medicine, Department of Histology and Embryology; 2Deptartment of Neurology, County Hospital Bjelovar; 3Department of Clinical Pathology "Ljudevit Jurak", Clinical Hospital "Sisters of Mercy' Introduction: Human foetal testis consists of two major parts: sex cords and interstitial compartment i.e. stroma. Sex cords had 40‐50 micrometers in diameter and are limited by several layers of peritubular cells. The cords are lined with pro‐spermatogonia and foetal Sertoli cells. Pro‐spermatogonia are considered as potential stem cells that could be used for regeneration of various organs, including the nervous tissue. Aim. The aim of our study was to determine if markers characteristic for central nervous system were expressed in pro‐spermatogonia. Materials & methods: A total of 39 human foetal testicles from 15 to 36 weeks of gestation were used in the study. The tissue samples were obtained during the routine paedopathological section of miscarried children during 1994 (approved by the Ethical Committee of Clinic for Gynaecology and Obstetrics, Clinical Hospital Centre "Zagreb"). The samples were fixed in Bouin's fluid, dehydrated and embedded in paraffin. Paraffin blocks were cut extensively in order to provide sections for hematoxylin and eosin (H+E), and immunohistochemical staining (THC). Monoclonal antibodies to NSE (1:100), GFAP (1:200), synaptophysin (1:25), chromogranin (1:100) and S‐100 (1:600) in combination with a appropriate En Vision kit (Dako, Glostrup, Denmark) were applied. Results & conclusion: Within sex cords, pro‐spermatogonia were recognized for their abundant and pale‐stained cytoplasm. Their nucleus was large, ovoid or round with abundant euchromatin and clumps of heterochromatin, predominantly associated with the nuclear envelope. Frequently, one or more prominent nucleoli could be noted within the nuclei of the cells. Sertoli cells were distinguished


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by their smaller size (when compared to pro‐spermatogonia), scanty cytoplasm and round or ovoid nucleus. IHC analysis pointed out a positive expression of NSE in the foetal testis interstitium as well as within sex cords. In the early stages of foetal development (15‐17 weeks of gestation, wg) elongated, spindle‐shaped cells were positive. In the period of 18‐36 wg, Leydig cells were positive for NSE (++). Moreover, in both of the above‐mentioned periods, a strong expression of NSE (++ +) was discovered in pro‐spermatogonia within sex cords, whereas foetal Sertoli and peritubular cells were negative. No positive results were obtained applying antibodies to S‐100, GFAP, synaptophysin and chromogranin. The expression of NSE in pro‐spermatogonia speaks in favour of multipotent character of these cells and capability to differentiate into various neural tissue forms. Teratomas and teratocarcinomas of testis that include parts of nervous tissue may be derived from afore‐mentioned cells. PP73 TRANSIENT MAP2‐POSITIVE CELLULAR BELT IN THE CORTICAL PLATE COINCIDE WITH BEGINNING OF THE AREAL SPECIFICATION. Jovanov‐Milošević N1, Petrović D2, Judaš M1, Kostović I1 1 Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Zagreb, Croatia, 2Department of Pathology, Clinic for Gynecology and Obstetrics, University Hospital Center Zagreb, Zagreb, Croatia We studied 32 specimens from Zagreb Neuroembryological Collection, from 15 postconceptual weeks (PCW) to 13 postnatal months (PM), stained by Nissl method and microtubule‐associated‐protein‐2 (MAP2) to disclose new cytoarchitectonical parameters of developing prefrontal cortex in human. First MAP2 reactivity in the cortical plate (CP) was seen as a band below marginal zone at 19 PCW. Between 21 and 26 PCW (when CP displayed incipient lamination), a belt of strongly MAP2‐positive pyramidal cells appeared in the middle of the entire prefrontal CP, but was most prominent in dorsolateral prefrontal region. MAP2‐reactive cells were also present in the marginal zone, deepest CP (prospective layer VI) and throughout the subplate zone (SP). This belt pattern in CP becomes less prominent at 27 PCW due to the increasing radial and tangential spread of prospective layer V pyramidal neurons and increase in MAP2‐staining of other cells in the CP. From 32 PCW to birth, MAP2‐positive belt gradually disappeared with concomitant development of areal differences in laminar staining (dorsolateral vs. orbitomedial). At 3 PM, MAP2‐positive cells are distributed throughout all cortical layers with remarkable first appearance of MAP2‐staining of layer III pyramidal neurons and staining in the remnants of the SP and in gyral white matter. At 13 PM, layer III and V pyramidal neurons displayed similar sizes and MAP2‐staining. In conclusion, period between 19‐26 PCW is important for cytoskeleton maturation of layer V pyramidal neurons. The transient MAP2‐positive belt characterizes developmental transition from regional to areal specification of the prefrontal cortex. This suggests that chemoarchitectonic differentiation of pyramidal neurons precedes the onset of frontal lobe functions. Presented on FENS FORUM 2008. Reviewed and updated for Third Croatian Congress of Neuroscience in Zadar, 2009. PP74 SPATIAL LEARNING AND MEMORY IN WISTAR‐ZAGREB 5HT RAT Mokrović Gordana, Jernej Branimir, Čičin‐Šain Lipa Laboratory of Neurochemistry and Molecular Neurobiology, Rudjer Bošković Institute, Bijenička 54, HR‐10000, Zagreb, Croatia Introduction: The serotonin (5‐hydroxytryptamine, 5HT) system has been implicated in various aspects of cognitive functions and central 5HT dysregulation contributes to deficits in learning and memory. By use of selective breeding, we have previously developed a rodent model, Wistar‐Zagreb 5HT rat, with two sublines constitutionally differing in peripheral (platelet) serotonin level and activity of 5HT transporter. We have further shown that besides in periphery, 5HT homeostasis is also changed in brains of 5HT sublines. In this study, we examined the relationship between altered brain 5HT functioning and cognitive performance in Wistar‐Zagreb 5HT rat sublines.


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Materials & Methods: Males and females from both sublines were tested for spatial learning and memory using Morris water maze task. The device consists of large circular pool with hidden escape platform, just below the water surface. During four days, animals are trained to find the platform using visual cues painted on the walls of the test room. Training consists of two sessions per day (intersession time 90 min), each including three 60 sec trials (intertrial time 15 sec). The time to find the platform is recorded and represents the measure of learning. On the test day (fifth day) the platform is removed from the water and the distance travelled and time spent in the quadrant where the platform was during learning, is recorded by computerized tracking system and represents the measure of memory. Results: In total of eight learning sessions, animals from the high‐5HT subline find the platform faster than those from the low‐5HT subline. In the memory test, animals from both sublines spent equal time in the quadrant where the platform was previously. Conclusion: This preliminary result shows possible differences in spatial learning abilities between sublines of WZ‐5HT rat, confirming the implication of serotonin in formation of spatial, hippocampus‐dependent, memory. PP75 QUALITATIVE AND QUANTITATIVE PROPERTIES OF DENDRITIC MORPHOLOGY COMPARING DIFFERENT SUBPOPULATIONS OF PRINCIPAL NEURONS IN INFRAGRANULAR LAYERS OF THE HUMAN PREFRONTAL CORTEX Paladin Ivan, Petanjek Zdravko Department of Neuroscience, Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Šalata 12, 10000 Zagreb, Croatia The principal neurons of the cerebral cortex are excitatory glutamatergic neurons projecting to other areas or brain regions. According to specific dendritic and cell body morphology they are defined as pyramidal cells. However, majority of principal cells in infragranular layers show some level of deflection compared to typical pyramidal morphology. Therefore, we define them as „modified" pyramidal cells. Morphological attributes of different modified pyramidal neuron subpopulations are not well defined. Using qualitative and quantitative morphological parameters of rapid Golgi impregnated neurons, the aim of this study was to define subpopulations of principal neurons in layers V and VI of the magnopyramidal Brodmann's area 9 in the dorsolateral part of human prefrontal cortex (PFC). Three subpopulations of principal neurons were defined: typical pyramidal cells in layer V, bipolar (fusiform) neurons in layer VI and transitory shaped cells (bipolar pyramids) at the border of these two layers. Quantitative analysis showed no differences between subpopulations in dendritic morphology and spine density of the apical dendrite and their side branches, but basal dendrites have less complex morphology on fusiform neurons. Data showed that principal neurons in infragranular layers of PFC are common to descriptions of other cortical regions and no areal specific cells were detected. It is assumed that morphological differences between subpopulations correspond to differences in functional properties. As expected, the evolutionary oldest cells located in the layer VI showed less complex dendritic morphology, but surprisingly this was only related to white matter oriented (basal) dendrites, and not to apical dendritic tree. This indicates that during evolution even the oldest neuronal elements should increase their complexity, to become functionally incorporated to extremely complex cortical network. Subpopulations of principal neurons are affected in numerous psychiatric and neurological diseases, and such studies can provide referent data for further research in this field. PP76 BLOCKADE OF 5‐HT1A RECEPTORS IN THE RAT PHRENIC NUCLEUS ATTENUATED RAPHE INDUCED ACTIVATION OF THE PHRENIC NERVE ACTIVITY Pecotić R1, Đogaš Z1, Valić Z2 and Valić M1

1Department of Neuroscience, University of Split School of Medicine, Split, Croatia, 2Department of Physiology, University of Split School of Medicine, Split, Croatia Introduction: Stimulation of the raphe pallidus nucleus produces facilitatory effects on respiratory activity. Numerous serotonergic projections from the raphe pallidus have been shown to terminate


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in the phrenic nucleus. This study was undertaken to examine the role of 5‐hydroxytryptamine 1A (5‐HT1 A) receptors in the phrenic nucleus on the excitatory response of the phrenic nerve activity elicited from the raphe pallidus. We hypothesized that blockade of 5‐HT1A receptors in the phrenic nucleus will attenuate raphe‐induced facilitation of the phrenic nerve. Materials and methods: Experiments were performed on urethane anaesthetized, spontaneously breathing, billateraly vagotomized adult male Sprague‐Dawley rats weighing 300‐330 g. Phrenic nerve was dissected using dorsal approach at the level of C5 nerve rootlet. The rats were placed in a prone position in a stereotaxic instrument. Occipital craniotomy exposed the dorsal surface of the brainstem. Triple‐barrel micropipette was inserted at coordinates targeting the raphe nuclei. The ejected solutions contained the vehicle, selective glutamate receptor agonist DL homocysteic acid (DLH, 10 mM; Sigma Aldrich, St Louis, USA), 0.9% saline, and India ink solution diluted in saline. Second micropipette was introduced in the phrenic motor nucleus ipsilaterally to the phrenic nerve from which electrical activity was recorded, at the level of C4 to C5 spinal segment. Micropipette tip was introduced vertically into the ventral portion of the right ventral horn 1.0‐1.2 mm lateral to the midline and 1.5 mm from the dorsal surface of the spinal cord. It contained the vehicle 0.9% saline, a selective antagonist of 5‐HT1A receptors WAY, N‐(2‐(4,2‐methoxyphenyl)‐l‐piperazinyl)ethyl)‐N‐2‐pyridinyl‐cyclohexane‐carboxamide maleate (WAY‐10063 5; 1 mM; Sigma Aldrich Chemie GmbH, Germany), and DLH. India ink was also used for labeling purpose. All drugs solutions were dissolved in 0.9% saline. Drug volume ejected was 20±5 nl. Results: Central administration of glutamate receptor agonist DLH into the raphe pallidus nucleus enhanced phrenic nerve activity in the spontaneously breathing anesthetized rats (55.2±6.8%, n=6, p<0.001). Microinjections of selective 5‐HT1A receptor antagonist, WAY‐100635, into the phrenic nucleus did not change phrenic nerve activity, but attenuated stimulatory effect on phrenic nerve activity elicited by DLH microinjections into the raphe nucleus. The raphe pallidus‐induced increase in the peak amplitude of the phrenic nerve activity was only 4.44±1.4% (n=6, p<0.05) after blockade of 5‐HT1A receptors in the PMN vs. 55.2±6.8% (n=6) prior to blockade of 5‐HT1A receptors in PMN. PP77 DEVELOPMENT OF MICROGLIA IN THE RHESUS MONKEY DURING MIDGESTATION Petanjek Zdravko, Džaja Domagoj, Milašin Goran Department of Neuroscience, Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Šalata 12, 10000 Zagreb, Croatia In this study we examined microglial development in telencephalon of cynomolgus monkey between embryonic day (E) 68 and E90 using the rabbit IgG and two monoclonal antibodies against the CD11b (OX42) and CD68 epitop, as a specific marker of primate microglia. Aggregates of closely packed oval to amoeba like or polymorph cells with patchy cytoplasmic staining (resembling the macrophage morphology) were regularly seen, through analyzed period, in following regions: (a) bellow the corpus callosum, (b) in the ventricular/subventricular zone of the temporal cortex, (c) in the internal capsule, (d) in the external capsule, and (e) in the ganglionic eminence. Smaller and not so densely packed aggregates were seen in the cingular bundle, rest of ventricular/subventricular telencephalic zone, fimbrial borders, as in the pia and choroid plexus. Numerous individual cells were seen in the intermediate zone and subplate. In addition, solitary cells with processes were also found around the regions of microglia aggregations. These cells were stained by CD11b and CD68 antibody but not by rabbit IgG. Majority of this dendritic cells resemble morphology of ramified microglia; most of them have one thick process bifurcating in brush like manner. On some of them few thinner processes could be distinguished and in many of them there was a patchy cytoplasmatic staining. The processes do not extend for distance longer than 50 microns with some varicosities. It is interesting that in addition to this dendritic like processes, on some of those cells one thin and longer, axon‐like process is present. In addition to this two population, resembling morphology of amoeboid and ramified microglia, lightly labeled cells that have uni‐bipolar morphology, typical for tangentionally migrating cells, and more differentiated, subplate neurons like cells were stained, too. Their density was highest around


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regions of microglia aggregates and decreased progressively in more distant positions, so that inside the cortical plate they were not observed. Thus, the highest density of the microglial cells is found on position of growing axon bundles and tangentially migrating cell routes. Therefore, we suggest that in the monkey brain during midgestation microglial cells might be involved in axon guidance and tangential migration. Our descriptive data also showed that these cells are at different stages of microglia differentiation, and that there might be interaction between microglia and immature/early born neurons. It seems that under influence of developing monkey brain environment, microglia can differentiate in to the cells that closely resemble neuron morphology. PP78 ORIGINS AND MIGRATORY ROUTES OF GABA‐ERGIC NEURONS IN THE PRIMATE CEREBRAL CORTEX Petanjek Zdravko, Kostović Ivica, Hladnik Ana, Esclapez Monique Department of Neuroscience, Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Šalata 12, 10000 Zagreb, Croatia γ‐aminobutyric acidergic (GABAergic) neurons that correspond mainly to local circuit neurons play a crucial role in regulating the activity of cortical neuronal networks and the complex interactions among principal neurons. These functions depend on a very heterogeneous population of neurons, whose number and diversity may increase during mammalian evolution. Interestingly, dysfunction and cell death of several specific types of GABAergic neurons is a hallmark of various psychiatric and neurological disorders such as schizophrenia, autism and epilepsies to which defect in genesis or migration of these neurons can be a predisposing factor. Understanding primate specific developmental mechanisms that regulate generation, migration and maturation of cortical GABA neurons, is a crucial step in elucidating such pathologies. However, there are few such studies in monkeys and humans, whereas they are numerous in lower mammals. In rodents, cortical GABAergic neurons are generated exclusively in the subcortical proliferative zone of the ventral telencephalon, the ganglionic eminence (GE), and migrate tangentially to the dorsal telencephalon to reach their target cortical layer. We reported that in contrast to rodents, in cynomolgus monkey cortical GABAergic neurons are generated massively in the proliferative zones of the dorsal telencephalon, in addition to the GE. This neurogenesis occurs very early during brain development in both dorsal and ventral telencephalon but with distinct temporal profiles. GABAergic neuron progenitors labeled for transcription factor Mashl and GABA synthesizing enzyme GAD65 were present mainly in the GE at embryonic days (E) 47‐55, and in the entire dorsal telencephalon at E64‐75. These progenitors within the dorsal telencephalon are generated locally rather than in the GE. In the human embryonic forebrain retroviral labeling in organotypic slice cultures also demonstrated the existence of two distinct lineages of neocortical GABAergic neurons. One lineage expresses Dlxl/2 and Mashl transcription factors, represents 65% of neocortical GABAergic neurons in humans, and originates from Mashl‐expressing progenitors of the neocortical proliferative zones of the dorsal telencephalon. The second lineage, characterized by the expression of Dlxl/2 but not Mashl, forms in human brain only around 35% of the GABAergic neurons and originates from the GE of the ventral forebrain, in comparison to rodent where all GABAergic neurons originate in GE. The dorsal telencephalic origin of cortical GABAergic neurons is a unique feature of human and non‐human primates and appears to be, in the primate lineage, an early evolutionary modification which provided increased number and variety of GABAergic neurons to an expanding neocortex. PP79 CHANGES OF OXIDATIVE STRESS PARAMETERS IN DIFFERENT RAT BRAIN REGIONS FOLLOWING TRAUMATIC BRAIN INJURY Pilipović K1, Župan Ž2, Frković V2, Dangubić B2, Mršić‐Pelčić J1, Šustić A2, Župan G1

1Department of Pharmacology, School of Medicine, University of Rijeka, Rijeka, Croatia, 2Department of Anesthesiology and Intensive Care Medicine, Clinical Hospital Center Rijeka, Rijeka, Croatia Introduction: Oxidative stress is considered to be important cause of secondary tissue damage following traumatic brain injury (TBI). Oxidative damage of cellular macromolecules, such as


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phospholipids, nucleic acids and proteins by reactive oxygen species, results in subsequent neuronal degeneration. Superoxide dismutase (SOD) and glutathione peroxidase (GSH‐Px) are main intracellular enzymes responsible for endogenous antioxidant defense of the brain regions affected by TBI. Therefore, the aim of our study was to evaluate the levels of lipid oxidative damage and the antioxidant enzymes' activities in different brain regions, at 24 h after the induction of TBI in rats. Materials & Methods: Experiments were performed on adult male Wistar rats. TBI of moderate severity was performed over the left parietal cortex using the lateral fluid percussion (LFP) brain injury model. Rats were sacrificed 24 h after the TBI induction. The brains were removed and the ipsilateral regions of interest (parietal cortex, hippocampus, thalamus, entorhinal cortex and cerebellum) were processed for spectrophotometric analyses of the thiobarbituric acid substances (TBARS) levels, as well as of the SOD and GSH‐Px activities. Sham‐operated animals were used as the control group. Results: TBI caused statistically significant increases of the TBARS levels in the ipsilateral cortex and hippocampus but it did not affect significantly the levels of the mentioned oxidative stress parameter in the thalamus, entorhinal cortex and cerebellum. GSH‐Px activities were significantly increased in the injured parietal cortex and hippocampus, while this enzyme's activities were not changed significantly in the other brain regions examined. SOD activities obtained from the injured animals were not significantly different in comparison to the levels measured in the sham‐operated animals in any brain region tested. Conclusion: Our results suggest significant increases of the levels of oxidative lipid damage and the GSH‐Px activities in the ipsilateral parietal cortex and hippocampus, at 24 h following TBI in rats. Thalamus, entorhinal cortex and cerebellum were not oxidatively altered in injured animals, at this time point in our experimental conditions. PP80 SERUM LIPID LEVELS IN ALZHEIMER'S DISEASE Presečki P1, Muck‐Šeler D2, Mimica N3,4, Pivac N2, Mustapić M2, Folnegović‐Šmalc V3,4

1Psychiatric Hospital Sveti Ivan, Jankomir 11, HR‐10 090 Zagreb, 2Ruđer Bošković Institute, Bijenička cesta 54, HR‐10000 Zagreb, Croatia, 3Psychiatric Hospital Vrapče, University Department of Psychiatry, Bolnička cesta 32, HR‐10090, 4School of Medicine, University of Zagreb, Šalata 3b, HR‐10000 Zagreb, Croatia Introduction: Alzheimer's disease (AD) is a complex and multifactorial neurodegenerative disorder. The role of lipids in the etiology and progress of AD is still unclear. Some studies have found that high lipid levels could be a risk factor for the cognitive decline in AD, whereas others have shown no association or even protective effects of high lipid levels in the development of the AD. Objective: To determine serum total cholesterol, high‐density‐lipoprotein cholesterol (HDL‐C), low‐density‐lipoprotein cholesterol (LDL‐C) and triglycerides (TG) levels in female patients with AD, subdivided according to the severity of disease, or the presence of psychotic features. Subjects and Methods: The study included 50 female patients with AD (mean age ± SD, 79.3 ± 8.3 years, range 5 3‐96 years), who met the diagnostic criteria of probable AD according to the NINDS‐ADRDA, ICD‐10 and DSM‐IV‐TR criteria. Cognitive impairment was evaluated using Mini Mental Status Examination (MMSE), and psychotic features by means of Neuropsychiatry Inventory. Patients were subdivided according to the MMSE scores into two groups: 19 patients in the middle (MMSE 18‐10) phase of AD and 31 patients in the late (MMSE 9‐0) phase of AD. Psychotic and non‐psychotic features were presented in 13 and 37 patients with AD, respectively. Serum lipid levels were determined by the enzymatic colour tests (total cholesterol, HDL‐C, and TG ), or by the enzymatic clearance assay (LDL‐C). Results: Patients in the late phase of AD had significantly lower serum total cholesterol (P<0.006, ANOVA) and LDL‐C (P<0.024) levels than patients in the middle stage of AD. There was no significant (P>0.05) difference in serum HDL‐C and TG levels, body mass index and age between patients with different severity of AD. The changes in serum total cholesterol, HDL‐C, LDL‐C and TG levels were not related to the presence of psychotic features. The small but significant positive correlations were


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found between MMSE scores and total cholesterol (r = 0.348; P<0.01), or LDL‐C (r= 0.291; P=0.04) levels in all AD patients. Conclusions: The results from our ongoing study support the presumption that lipid profile might be connected with the etiology and progress of AD. In addition, our results showed the association between low total cholesterol and LDL‐C levels and cognitive decline in patients with AD, suggesting a protective effect of high lipid levels on cognition in patients with AD. Further studies are needed to confirm that lipid profile could be a biological marker for the progress of the AD. PP81 SEROTONIN (5HT) METABOLISM IN THE BRAIN OF WISTAR‐ZAGREB 5HT RATS Štefulj Jasminka1, Čičin‐Šain Lipa1, Bokulić Zvonimir1, Mokrović Gordana1, Bordukalo‐Nikšić Tatjana1, Orešković Darko1, Živin Marko2, Jernej Branimir1 1Laboratory of Neurochemistry and Molecular Neurobiology, Ruđer Bošković Institute, Bijenička 54, HR‐10000, Zagreb, Croatia, 2Medical Faculty, University of Ljubljana, Slovenia Introduction: Sublines of Wistar‐Zagreb 5HT rats (WZ‐5HT rats), characterized by constitutionally upregulated (high‐5HT subline) or downregulated (low‐5HT subline) platelet 5HT transporter (5HTT), display differences in the behavioural traits associated with 5HT neurotransmission, as well as differences in the 5HTT‐related neuropharmacological response. On the other hand, no basal differences between the sublines were found in the expression and/or density of 5HTT and several 5HT receptors in selected brain regions. In attempt to characterise molecular mechanisms underlying observed changes in the brain functioning, here we have examined potential differences between 5HT sublines in the brain 5HT metabolism. Methods: Brain levels of 5HT and its major metabolite 5‐hydroxyindoleacetic acid (5HIAA) were examined by HPLC‐ED. Brain expression of 5HT synthesising enzyme tryptophan hydroxylase 2 (TPH‐2) was assessed by western blot, real‐time RT‐PCR and in situ hybridisation. 5HT degrading enzymes monoamineoxidase A (MAO‐A) and B (MAO‐B) were analysed functionally by full kinetic measurement and at the level of mRNA by conventional RT‐PCR. Results: Animals from high‐5HT subline displayed non‐significant trend towards reduction of 5HT and elevation of 5HIAA levels in multiple brain regions, resulting in a significantly higher (‐10%, p<0.05) metabolic ratio (5HIAA/5HT) in the most regions examined. TPH‐2 protein levels in the raphe nuclei region were for about 30% lower in high‐5HT rats as compared to low‐5HT rats, although the difference did not reach the level of statistical significance (p=0.1242). The same direction of changes, but also non‐significant, was observed for TPH‐2 mRNA levels, as assessed by real‐time RT‐PCR and in situ hybridisation. On the other hand, MAO‐B mRNA levels in the cortical region were significantly (p<0.05), although slightly (‐10%), increased in low‐ as compared to high‐5HT subline, while no alternations were found in the raphe region. There were no differences between the sublines in the levels of MAO‐A mRNA, as well as in the kinetic properties (Vmax, Km) of MAO‐A and MAO‐B isoenzymes, as measured in the whole brain homogenates. Conclusion: Results indicate that animals from the high‐5HT subline have slightly increased brain 5HT turnover as compared to animals from low‐5HT subline. In certain brain regions, indicated changes in the 5HT turnover could possibly be attributed to the alternations in the expression of 5HT metabolic enzymes. PP82 DIFFERENT EXPERIMENTAL PROTOCOLS AND PHRENIC LONG‐TERM FACILITATION Valić M1, Pecotić R1, Pavlinac I1, Valić Z2 and Đogaš Z1 1Department of Neuroscience, University of Split School of Medicine, Split, Croatia, 2Department of Physiology, University of Split School of Medicine, Split, Croatia Introduction: Long term facilitation (LTF) represents a form of neural plasticity that persists long after exposures to episodic hypoxia have ceased. LTF stimulation protocol employs wide variety of hypoxic episodes at different 02 levels that could affect the elicitation, magnitude and duration of phrenic LTF in anesthetized rats. To demonstrate importance of proper stimulation, the present study examined the effects of two different experimental protocols in the LTF induction.


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Materials and methods: The phrenic nerve activity (PNA) was recorded in 10 adult, male, Sprague‐Dawley rats, urethane anesthetized, bilaterally vagotomized, paralyzed and mechanically ventilated. Femoral arteries and veins were cannulated for blood pressure monitoring, obtaining blood samples, and intravenous drugs administration. The phrenic nerve recordings monitored central respiratory activity. Phrenic nerve was dissected using dorsal approach at the level of C5 nerve rootlet, mounted on bipolar silver wire electrodes. The rats were placed in a prone position in a stereotaxic instrument. The animals were divided in 2 groups. One group (n=5) was exposed to five hypoxias that lasted for 3 minutes each (FiO2=0.09), separated by 3 minutes of hyperoxia (FiO2=0.5). Another group was exposed to three hypoxias that lasted for 5 minutes each (FiO2=0.09), separated by 5 minutes of hyperoxia (FiO2=0.5). Peak phrenic nerve amplitude (PNA), burst frequency (f), inspiratory time duration (Ti), expiratory time duration (Te) and total respiratory cycle duration (Ttot) were analyzed during the first hypoxia and 15, 30, and 60 minutes after the final hypoxic episode, and compared to the baseline values prior to the first hypoxic episode. Results: Isocapnia was successfully maintained throughout the protocol. There was a significant increase of the peak phrenic nerve amplitude (66.7 ± 8.6%, p<0.05) in group of animals that underwent 5 times 3 minutes hypoxic exposures 60 minute after the last hypoxic episode compared to the baseline values. On the contrary, in the second group of animals that underwent 3 times 5 minutes hypoxic exposure, no significant changes of PNA were recorded at any time point after the last hypoxic episode. AIH did not elicit significant changes in the phrenic burst frequency, as well as in breathing rhythm parameters (Ti, Te, Ttot) in both groups at all 3 time points after the last hypoxic episode. Conclusion: This study demonstrates that protocols with different number and duration of hypoxic episodes differently affect the elicitation of the phrenic LTF in urethane anesthetized rats. Protocol of 5 times 3 minutes intermittent hypoxic exposure evokes phrenic LTF whereas another protocol (3 times 5 minutes) failed to elict phrenic LTF. PP83 DYNAMICS OF THE VOLUME CHANGES OF THE TRANSIENT CELLULAR ZONES IN THE DEVELOPING HUMAN CEREBRAL WALL Lana Vasung1, Alan Evans2, Ivica Kostović1 1Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Croatia, 2Montreal Neurological Institute, Brain Imaging Centre, McGill, Montreal, Quebec, Canada Fetal cerebral wall is composed of the transient cellular zones witch are sites of neurogenetic events (Bystron et al. 2009). These transient cellular zones can be well delineated on postmortem human brain using MRI (Kostovic et al. Cereb Cortex 2002). In order to obtain quantitative data on their development we have reconstructed and analyzed their dynamic volume changes as prospective indicators of intensities of their predominant neurogenetic events during prenatal development. Tl weighted MRI images of 17ex vivo fetal brains, aged 13‐40 PCW, were acquired on 3T Siemens scanner with voxel resolution ranging from 0.25‐0.4 mm. We have extracted cortical plate (CP) surface and subplate surface using modified Montreal Neurological Institute toolbox. For reconstruction and volume measurement of transient fetal laminae we have used Analyze 7.0 (Mayo Clinic) software. In the early fetal life, from 9‐13 post‐conceptual weeks (PCW), fetal cerebrum is organized in thick and densely packed proliferative cellular zones (ventricular/subventricular zone and ganglionic eminence). During this period proliferative zones occupy 1/3 (32 %) of total cerebral volume. The fetal phase (13‐24 PCW) is characterized by the appearance and exponential growth of other fetal zones which are sites of intensive fiber ingrowth, synaptic development and neuronal differentiation: (1) intermediate zone, (2) subplate zone and (3) CP. Although proliferative fetal zones during this phase show an increase of volume by the factor of3.8 the 41.3% of the cerebral volume is occupied by subplate zone at the end of this phase. Finally, during the preterm phase (24‐35 PCW) neurogenetic events of cerebral development are reflected trough (1) existence of transient, synapse rich subplate zone, (2) growth and initial myelization of fetal white matter and (3) establishment of adult like cortical laminar pattern. During this phase proliferative zones are showing decrease by a factor of 4,4 and CP and subplate increases by factor of5.By the end of this phase CP becomes the


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thickest lamina showing an exponential volume growth parallel with gradual decrease of subplate. In conclusion, our preliminary results support hypothesis that quantitative MRI parameters of transient fetal laminae can serve as a valuable pointers of intensity of underlying fetal neurogenetic events. Supported by UKF. PP84 DISTRIBUTION OF MAJOR BRAIN GANGLIOSIDES IN OLFACTORY TRACT OF FROGS Barbara Viljetić1, Ivan Večeslav Degmečić2, Vinko Krajina6, Tomislav Bogdanović3, Ana Mojsović4, Domagoj Đikić5, Katarina Vajn6, Marija Heffer‐Lauc6 1Department of Chemistry and Biochemistry, School of Medicine, University J. J. Strossmayer, J. Huttlera 4, Osijek, Croatia,2Animal Facility, School of Medicine, University J. J. Strossmayer, J. Huttlera 4, Osijek, Croatia, 3Department of Biology, University J. J. Strossmayer, Gajev trg 5 , Osijek, 4School of Nursing, Mlinarska cesta 38, Zagreb, 5Department of Animal Physiology, Faculty of Science, University of Zagreb, Rooseveltov trg 6, Zagreb, 6Department of Medical Biology, School of Medicine, University J. J. Strossmayer, J. Huttlera 4, Osijek, Croatia Gangliosides are major cell‐surface determinants in central nervous system of vertebrates, found both in neuronal and glial cell membranes. Together with cholesterol and glycosylphosphatidylinositol (GPI) – anchored proteins, gangliosides are involved in organization within plasma membrane microdomains. Based on biochemical studies, frog brain was previously described as having low quantities of gangliosides and their distribution pattern in specific brain regions was unknown. Using highly specific monoclonal antibodies raised against four major gangliosides (GM1, GD1a, GD1b and GT1b), we examined the distribution of these molecules in central nervous system of five different species of frogs (Pelobates fuscus, Rana esculenta, Rana temporaria, Bufo bufo and Bufo viridis). We had also studied the distribution of myelin‐ associated glycoprotein (MAG), an inhibitor of axonal regeneration, which is a ligand for gangliosides GD1a and GT1b. Our results show that ganglioside GD1a is expressed in neurons of olfactory bulb in all studied animals, except Pelobates fuscus. In the brain of Rana sp., GD1a is expressed in the entire olfactory pathway, from olfactory bulbs toward amygdala. We had also found that Rana sp. expresses GD1b in same neurons and fibers as GD1a, but just if the animal is kept at ambient temperature above 15°C. Since both gangliosides (GD1a/GD1b) are present in same neurons, we presume that they assist in organization of plasma membrane microdomains important in signaling pathways of olfaction. In frogs, olfaction is more important than vision or any other sensory clue, because it affects seasonal and mating behavior. Furthermore, we found that all other myelinated pathways in frogs express MAG, but do not express GD1a or GD1b. Ganglioside GD1a is also not expressed in visual pathway, which is known to regenerate after its lesion. Keywords: Gangliosides, Myelin‐Associated Glycoprotein (MAG), Membrane Microdomains, Amphibians PP85 DENDRITIC REORGANIZATION OF GRANULE CELLS IN THE FASCIA DENTATA OF THE THY1‐GFP MOUSE FOLLOWING ENTORHINAL CORTEX LESION Vukšić Mario1,2, Del Turco Domenico1, Muller Christian M.1, Deller Thomas1 1Institute of Clinical Neuroanatomy, J. W. Goethe‐University, Theodor‐Stern‐Kai 7, D‐60590 Frankfurt/ Main, Germany,2Croatian Institute for Brain Research, University of Zagreb, Šalata 12, HR‐10000 Zagreb, Croatia Introduction: Neurons denervated after brain injury remodel their dendritic tree. Although this phenomenon has been described many years ago, neither the precise dynamics of the structural changes nor the underlying molecular mechanisms have yet been unraveled. The entorhinal cortex lesion (ECL) model is one of the classical models for the analysis of denervation‐induced neuronal changes in the CNS. However, most of the data stem from ECL in rats and data on the reorganization


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of neurons in the denervated mouse fascia dentata are scarce. Since this limits the use of genetic tools, we have analyzed the dendritic reorganization of denervated granule cells in mice. Materials & Methods: Thy 1‐GFP mice bred on a C57BL/6 background were used in these studies. Entorhino‐hippocampal fibers were transected using a wire knife and animals survived 3, 7, 10, 30, 90 and 180 days post lesion (dpi). Denervated GFP‐labeled granule cells located in the dorsal fascia dentata were 3 D‐reconstructed in their entirety using confocal microscopy and Neurolucida system. Results: Our morphometric analysis revealed that ECL results in a profound atrophy of denervated mouse granule cells. Dendritic length and the number of dendritic segments progressively decreased and reached a minimum around 90dpl. Dendrites located within the denervated portion of the molecular layer were predominately affected . By 180dpl a partial recovery of dendritic length but not of dendritic segments was observed. Spine density showed a similar decrease with a minimum around 90dpl (70% of all spines lost; spine density decreased to 50% of control values). By 180dpl a partial recovery in spine number and density was seen. Using intracellular fillings of neurons in fixed hippocampal slices we found no significant morphological differences between GFP‐positive and GFP‐negative granule cells upon denervation. Conclusion: This study provides normative data on the ECL model in mice and reveals that granule cell atrophy in mice following denervation is more severe than in rats. Entorhinal denervation in mice results in permanent dendritic atrophy. In spite of sprouting, little dendritic reconstruction occurs, suggesting that sprouting may halt the atrophic degeneration but can not reverse it.

P8 NEURODEVELOPMENTAL BASIS OF COGNITIVE, MENTAL AND NEUROLOGICAL DISORDERS

PP86 LENTICULOSTRIATAL VASCULOPATHY (LSV) ‐ A MARKER FOR CONGENITAL CMV INFECTION? Đuranović Vlasta, Mejaški‐Bošnjak Vlatka, Lujić Lucija, Krakar Goran, Gojmerac Tomislav University of Zagreb, Children's Hospital Zagreb, Klaićeva 16, Zagreb Lenticulostriatal vasculopathy (LSV) is an ultrasonographic (US) brain lesion which appears as branched candlestick ‐ shaped echogenicities or hyperechogenic punctuations located within the basal ganglia (BG) and the thalamus. It has been reported in 0.27‐2.5% live neonates, 5.1‐32% preterm infants and 1.9‐5,8% ill neonates, the most often in those with congenital cytomegalovirus (CMV) infection. Congenital CMV infection is an important cause of morbidity and mortality in neonates and infants. Brain ultrasonography (US) showed: abnormal echogenicity of the parenchyma, echogenic sulci and gyri, ventricular dilatation, complications and sequellae of the infection (ventriculitis, abscesses, hydrocephalus and calcifications), disturbances of neuronal migration, cerebellar hypoplasia, delays in myelination, SE cysts and LSV. LSV may be caused by a variety of conditions including infection, haemorrhage, hypoxic‐ischemic insult, hypoglycaemia, infarction, calcifications, vascular lesions, metabolic disorders or chromosomal abnormalities. Transfontanellar Color Duplex Doppler (TFCD) imaging confirms the vascular origin of these lesions with pulsatile flows as an arterial spectrum. It revealed in the first months of life, but after that time TFCD showed no more flow within the hyperechogenicities, suggesting true calcifications. LSV can be graduated as unilateral or bilateral, isolated or with coexistent abnormalities, grading to minor, moderate and major LSV. The aim of this work is to present TFCD findings in neonates and infants with risk for development of neuromotor disabilities. All infants have US findings ofLSV in BG and thalamus. TFCD showed a flow inside during the first few months of life, but with no flow within the echogenicities at the end of the first year. We would like to perform an etiology and graduation ofLSV with implications for neurodevelopmental outcome, especially in infants with congenital CMV infection.


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PP87 CHARACTERISTICS OF HEARING LOSS IN UNILATERAL CLEFT LIP AND PALATE – INFLUENCE ON COMMUNICATION Handžić Jadranka, Vladika Ivan, Šimunčić Veselić Anamarija, Bura Miljenko, Radić Božo University Clinical Hospital Center Zagreb, Medical School, Department for Otolaryngology, Head and Neck Surgery Introduction: Conductive hearing loss accompanied by otitis media with effusion (OME) is usual finding in cleft palate population. Natural history of OME is still unclear and often described as multi‐factorial. Objective is to examine relationship between hearing levels, audiometric frequencies aging and ear side in left (UCLP) (L) and right (UCLP) (r) unilateral cleft lip and palate (UCLP) children. Materials and Methods: We selected UCLP children from general population during their visit to outpatient audiology department and their charts analyzed retrospectively. Total sample of UCLP children (n=101, median age, 6 yr) and subgroups of those with cleft on left (UCLP) (L) (n=74, median age 6 yr) and right side (UCLP) (R) (n=27, median age 7 yr). Tonal audiometry established average hearing levels (AHL) across frequencies from 250 Hz to 4 KHz , and their median hearing level (MHL) for left and right ears of study groups respectively .Ears of the non‐cleft side were controls to the ears of the cleft side. Results: Left UCLP (L) ears usually had moderate conductive loss and had decrease in frequency with aging. Frequency of NHL did not increase with aging. At 1‐3 yr highest median hearing level (MHL=32,5dB) had frequencies of 1, 2, and 4 kHz. Improvement of hearing level found at age 8‐12 yr at 0,5, 1, 2 KHz and average hearing level (AHL) Right UCLP (L) ears had highest median hearing level at 0,5 KHz. Improvement of hearing loss found at 4‐7 yr for all frequencies except 0, 25 KHz .They showed age related increase in frequency of normal hearing ears. Left UCLP (R) patients showed lower MHL at 0, 25 KHz at 1‐3 yr and 4‐7 yr with significant improvement until age 8‐12yr.Right UCLP (R) ears showed no significant differences for the same parameters. Conclusion: Hearing loss in UCLP children showed left cleft side and age dependence. PP88 INFANTILE SPASMS IN CHILDREN WITH DOWN SYNDROME Lujić L, Mejaški Bošnjak V, Delin S, Đuranović V, Krakar G Children's Hospital Zagreb, Medical School,University of Zagreb Pediatic department, General Hospital Zadar, Croatia Down syndrome (DS) is the most frequent genetic cause of mental retardation. In children with DS, beside typical phenotypic characteristics and mental retardation, there is higher incidence rate of congenital hypothyreosis, deafness, congenital heart disease, immune deficit and malignant diseases than in general population. According to available studies, seizures occur in 5‐13% of those children, with higher incidence rates among children younger than 12 months. Infantile spasms (IS) are most common type of seizures and are usually well controlled by the administration of corticosteroids and other antiepileptic drugs. Tendency for the appearance of seizures is in connection with structural abnormalities ofbrain development in DS, but the exact mechanism is yet to be determined. Evidence suggest a possible interaction among excessive neuronal excitability caused by alteration in neuronal membrane channels, dysgenesis of dendritic spines and abnormal inhibitory transmission caused by a 20‐50% decrease in the number of small inhibitory interneurons in cortical layers II and IV. Other factors such as perinatal asphyxia and infection as well as postnatal hypoxia caused by congenital heart disease also increase frequency of seizures. We present eleven children (9 male and 2 female) with DS and infantile spasms, who were treated in Children's Hospital Zagreb from January 2000 to January 2009, all with regular 21 trisomy. Nine of those eleven children had congenital heart disease and one of them had to undergo heart surgery because of A‐V canalis communis. The onset of infantile spasms was at the age of 5‐10.5 months in ten children and one child had the onset at the age of 16 months. Only one child had perinatal risk factors for development oflS. EEG findings in all DS children were consistent with hypsarrhythmia. Brain image analysis (intracranial ultrasound, CT and/or MRI) demonstrated diffuse cortical atrophy in nine children. Intracranial ultrasound findings were normal in two children. Initial peroral anticonvulsive therapy (usually valproate) was inefficient


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in 10/11 children. After treatment with ACTH infantile spasms ceased between day 7 and 20 in eight children and after 28 days in two more children. Hypsarrhythmia disappeared in all children. During the follow‐up time (from 2 years and 2 months until 8 years and 10 months) none of those patients developed any other type of seizures. Also there was no significant epileptogenic activity shown in EEG. In four children anticonvulsive therapy was withdrawn in the age of 4 years and 2 months to 5 years, including boy who died ofheart disease in at the age of 4 years and 3 months. PP89 BRAIN MALFORMATIONS IN CHILDREN WITH CONGENITAL CYTOMEGALOVIRUS Mejaški‐Bošnjak V, Krakar G, Đuranović V, Rakvin I, Delin S Children's Hospital Zagreb, Medical School,University of Zagreb Pediatic department, General Hospital Zadar, Croatia Congenital cytomegalovirulovirus (CMV) infection is the most common transplacentary transmitted viral infection, occuring in approximately 1% oflive born neonates. CMV may cause multiorgan affection but organ specific disease, with the most serious and permanent sequelae to central nervous system ( CNS). Advanced diagnostic methods in particularly laboratory and neuroimaging contribute to better recognition of congenital CMV infection. Congenital CMV infection may disturbe neurogenesis of CNS. Structural abnormalities of the CNS are related to the timing of affected neurogenetic processes. Early‐onset infection before 16‐18 weeks of gestation causes severe disorder of neuronal proliferation, migration and cerebellar hypoplasia. We present 7 children with brain malformations and proven congenital cytomegalovirus infection. In all children congenital CMV infection was confirmed by specific serology, PCR for CMV DNA and/or virus urine excretion. All children underwent MRI, and 4 of them cranial CT. All 7 children have complex brain malformations : lissencephaly in 2 children, in one of them accompanied by cerebellar hypoplasia , parenchymal calcification and callosal hypoplasia. Another 4 children have widespread cortical dysgenesis (pachygyria/polymicrogyria), accompanied by cerebellar hypoplasia callosal hypoplasia parenchymal cysts and calcification.Microcephaly was present in all children, reflecting severely disturbed brain development. Brain malformations caused by congenital CMV infection are severe and complex affecting cortical organisation, neurogenesis of cerebellum, parenchymal cysts and callosal hypoplasia. Calcifications are present in 4 out 7 children, which was better visualized by cranial CT. Severe and complex brain malformations causing cortical dysgenesis, cerebelar hypoplasia indicate an early‐onset infection. PP90 EARLY INTERVENTION IN INFANTS AT HIGH RISK FOR A NEURODEVELOPMENTAL DISORDERS Pinjatela Renata, Joković Oreb Ines Faculty of Education and Rehabilitation Sciences, University of Zagreb Re/habilitation of children with neurodevelopment risk represents a complex practical model which demands directed scientific approach. Existence of neurodevelopment risk, although no necessary connected with incidence of development disturbances, often deranges good interaction between parents and child and is particularly expressed in the first year of the child's life. Problem becomes more pronounced if neurodevelopment disturbance is diagnosed. Entering the family that has a child with expressed development disturbance in his/hers first year, a number of problems with which parents have to cope during that very important and valuable period are perceived. The most important is adaptation to the new situation. In the Republic of Croatia early intervention programs are directed primarily more to the child deficit and less to the parents, family and coordination between members of the expert team. This results with longer diagnose process accompanied with the uncertainty and exhaustion of parents. The implementation of project "Integration Development Programs for Early Intervention" started in 2005 in collaboration of Centre for Rehabilitation of the Faculty of Education and Rehabilitation of the University in Zagreb and Ministry of Family, Veterans' Affairs and Intergenerational Solidarity of the Republic of Croatia. Incentive for this project came from the needs of re/habilitation practice which is characterized with the insufficient representation and coordination of integration programs


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in the early intervention procedures for children with neurodevelopment risk and insufficient education and support of the family in the earliest period of child development. Project is contribution to the Priority activities for the child welfare derived from the National program acting for children in the Republic of Croatia and has the aim to develop protection measures for the children born with the risk factors. Program will contribute to the preventive and therapeutic education‐rehabilitation activity. In project participate rehabilitators‐ defectologist, speech pathologists, physicians of different specialist, psychologists, physiotherapists, contributing thus to the transdisciplinary characteristic of the research in the area of neurodevelopment re/habilitation. Furthermore, the results of the project will also contribute to the further development of included disciplines. Key words: neurorisk/neurodevelopmental disorders, early intervention programs, family

P9 SPECIAL SESSION – CASE REPORTS

PP91 DIFFERENTIAL DIAGNOSTIC RELEVANCE OF HIGH RESOLUTION MAGNETIC RESONANCE IN PATIENTS WITH PROBABLE EARLY MULTIPLE SYSTEM ATROPHY (MSA) – A CASE REPORT K. Bačić Baronica1, G. Ivkić2, D. Ozretić3 1University department of neurology, General Hospital „Sveti Duh”, Zagreb, Croatia,2Croatian Institute for Brain research, School of Medicine, University of Zagreb, Diagnostic Center “NEURON”, Zagreb, Croatia,3Department of diagnostic and interventional Radiology, University Hospital Zagreb, Zagreb Background. Multiple system atrophy (MSA) is sporadic, progressive neurodegenerative disorder characterized by combination of three possible group of dysfunctions: autonomic dysfunctions (MSA‐A), Parkinsonism (MSA‐P) and cerebellar dysfunctions (MSA‐C). Parkinsonism is present in the majority of all patients (80%), and is associated with rapid progression of disease. Early in the course of the disease autonomic dysfunctions are present in approximately 40% of patients (later in almost all patients), while the domination of cerebellar symptoms is present in 20% of all patients. Brain magnetic resonance imaging (MRI) is the method with potential for objective quantification of main abnormalities that are critical in the differential diagnosis of MSA and related disorders. Case report. We present a case of 60‐year old patient with history of progression (predominant cerebellar) symptoms, followed by the extrapyramidal symptoms and few mild symptoms of autonomic dysfunction, lasting six months. Patient was completely clinically investigated, including the complete neurological examination, psychological testing, cardiological evaluation (24 hours blood pressure monitoring,, 24 hours ECG heart rate variability), laboratory and radiological imaging (1.5T MRI, and additionally the 3T MRI). Results. On initially performed 1.5T MRI, the most of neurodegenerative features of brain stem, cerebellum and basal ganglia (atrophy) were well illustrated, while relative typical MR features of MSA (“Hot‐cross bun”) were detected only after MR imaging on higher resolution (3T) device. The level of quantification power of most neurodegenerative features was also much higher. Discussion. In relative short period from the onset of the first symptoms, it is possible that the signs of autonomic dysfunction are not fully present and that the leading symptoms are cerebellar or/and extrapyramidal. This can be a differential diagnostic problem in establishment of the diagnosis of MSA according to existing algorithm based on clinical presentation. The “High‐power” MRI, with special estimated sequences (T2 and FLAIR) in correlation with clinical picture, represents, in this (early) phase, a crucial diagnostic tool for establishing the definitive (clinical) diagnosis of MSA.


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PP92 DEPRESSIVE EPIZODE IN AN ADOLESCENT GIRL WITH TUBEROSE SCLEROSIS COMPLEX Boričević Maršanić Vlatka Psychiatric hospital for children and youth, Kukuljevićeva 11, 10 000 Zagreb Tuberose sclerosis complex (TSC) is a multisystem genetic disorder caused by mutations in the tumor suppressor genes, TSC1 or TSC2. It is associated with the formation of multiple hamartias and hamartomas throughout the body, but particularly in the brain. There is a wide range of structural and functional central nervous system manifestations. Approximately half of individuals diagnosed with TSC present with cognitive and behavioral problems. During adolescence, a significant number of individuals with TSC develop severe affective symptoms. A case of the first severe depressive epizode associated with TSC in an adolescent girl is reported. Possible mechanisms responsible for the secondary mood disorder associated with TSC are discussed. Key words: tuberose sclerosis complex, depression, adolescence, cognitivne behavioral therapy PP93 CEREBELLAR GLIOBLASTOMA IN ELDERY PERSON ‐ CASE REPORT Božić Boris, Rotim Krešimir, Kčira‐Fideršek Vječeslav, Krpina Hrvoje, Čupić Hrvoje Department of Neurosurgery, University Hospital "Sestre milosrdnice", Zagreb, Croatia, Department of Patology "Ljudevit Jurak", University Hospital "Sestre milosrdnice", Zagreb, Croatia Gliomas are the most frequent primary brain tumors. They arise from neuroepithelial cells and consist 40% of all intracranial tumors and 78% of all malignant tumors of central nervous system. They are divided in three basic groups: astrocytic tumors, oligodendroglial tumors and ependymal tumors. World Health Organization has divided in four groups of malignancy. The most malignant ones are the glioblastomas, grade IV tumors. They consist 12 to 15 % of all intracranial tumors, and can develop from low grade astrocytomas or arise "de novo". Glioblastoma can appear in any age group, but shows more often in patients above 50 years of age. It is usually found above tentorium, usually in cerebral hemispheres white matter. Inftarentorial glioblasomas are rare; so far only 33 cases of glioblastomas have been reviewed in literature. 65 year old male patient was admitted at Department of Neurology at University hospital "Sestre milordnice". His symptoms included vertigo, vomiting, ataxia and headache. He started to deteriorate fast so MSCT of the brain showed hydrocephalus and posterior fossa expansive lesion measuring 34 x 33 x 52 mm with compression and dislocation of the fourth ventricle. Patient was urgently transferred to Department of Neurosurgery where external ventricular drainage was placed and measuring pressure of 600 mm of water. Postoperatively the patient's condition improved and stabilized. Patient underwent another surgical procedure; the medial suboccipital craniectomy was performed and the tumor was microsurgical removed. Recovery was very good, early physical therapy was performed. Postoperative CT scan showed no tumor residua. Patohistological findings have verified that the tumor is Glioblastoma multiforme. Four weeks later the patient underwent 30 fractions of radiotherapy, total of 60 Gy, and chemotherapy with temozolamide treatment. One year after surgery the patient returned at his job as a musician and composer. PP94 MAGNETIC RESONANCE FINDINGS IN A NEONATE WITH NONKETOTIC HYPERGLYCINEMIA ‐ CASE REPORT Čuljat M1; Benjak V2; Dasović‐Buljević A2; Ozretić D3; Fumić K4; Barić I2 1Croatian Institute for Brain Research, School of Medicine, University of Zagreb; 2Department of Pediatrics, Clinical Hospital Center Zagreb; 3Department of Radiology, Clinical Hospital Center Zagreb; 4Laboratory for Diagnosis of Metabolic Diseases, Clinical Hospital Center Zagreb INTRODUCTION: This report presents brain imaging and spectroscopy data in a 7‐day‐old neonate with a later confirmed classical neonatal form of nonketotic hyperglycinemia (NKH), an autosomal recessive metabolic disorder characterized by accumulation of glycine in cerebrospinal fluid, plasma and urine. Her CSF glycine was 235 μmol/L, and plasma glycine 875 μmol /L, giving the ratio of 0.27 (normal <0.05). We detailed the changes seen on conventional magnetic resonance imaging (MRI), diffusion weighted imaging (DWI) and magnetic resonance spectroscopy (MRS). At that time‐point


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her daily protein intake was 1.7 g/kg of b.w. She received no therapy that could influence spectroscopy findings. Changes on MRI, such as increased T2 signal intensity, diffusion parameters showing restricted diffusion and an additional peak on MRS, were previously described in older children. To our knowledge, ours is the only report of such complete analysis of magnetic resonance imaging parameters in a patient with NKH at neonatal age. METHODS AND RESULTS: Conventional T1‐weighted imaging of our patient showed a regular organization of gyri and sulci, with a slightly pronounced diffuse reduction of the telencephalic gray and white matter, with a reduction of the midsagittal area of the corpus callosum. Hypoplasia of the caudal vermis was found, as well. On T2‐weighted images (TR=6000; TE=91; ETL=17) high‐signal lesions were found in the posterior limbs of internal capsules, tegmentum of the brain stem and in middle and inferior cerebellar peduncles. The same regions had increased signal intensity on diffusion weighted imaging (TR=7600; TE=111; b=1000 s/mm2) combined with low ADC values, indicating restricted diffusion. However, normal fractional anisotropy was present in these areas, indicating preserved axonal integrity. Single‐voxel proton spectroscopy (TE=2000 ms; TE=135 ms) showed a clear peak at +3,6 ppm, corresponding to glycine, in the frontal and parieto‐occipital white matter, the basal ganglia and the cerebellar hemisphere. DISCUSION: MR brain imaging shows clear pathological changes in NKH children. The findings of a 7‐day‐old neonate are consistent with the previous reports in older children with NKH, confirming that the pathological changes typical for NKH can be seen in the first postnatal week. The need of a detailed clinical work‐up and an early MRI examination in these children is emphasized by the existence of a classical and transient variant of the disease, with drastically different outcomes, where MR imaging might aid in distinguishing the two forms and in giving the prognosis. PP95 ACUTE LONGITUDINALLY EXTENSIVE TRANSVERSE MYELITIS IN SIX YEAR OLD CHILD ‐ CASE REPORT Delin S1, K Pavešić K1,Tešović G2, Knezović I2, Miše B2

1General Hospital Zadar, Zadar, 2University Hospital for Infectious Diseases "Dr. Fran Mihaljević", Zagreb Acute transverse myelitis(ATM) is a rare disease in childhood an adolescence age. In the United States ATM has an incidence of 1‐4 new cases per millon people per year, but there is no specific evaluation of pediatric incidence ATM is characterized by bilateral spinal cord dysfunction presenting as lower extremity weakness with sensory simptoms and bladder dysfunction. It typically manifests over a period of hours to one week. A hystory of preceding viral infection accompanied by fever and malasie is documented in 60% of cases. Several viruses have been implicated including the Ebstein‐Barr virus and cytomegalovirus, herpes simplex, influenza, rubella, mumps and varicella viruses. Spinal magnetic resonance imaging (MRI), electrophysiological tests and cerebrospinal fluid analysis are performed to diagnose ATM and exclude other treatable conditions. ATM can be caused by a number of disorders including trauma, structural and space occupying lesion, vascular malformations, occlusive vasculitic disorders causing infraction of the spinal cord. ATM can be the initial presentation of acute lymphoblastic leukemia. Guillain Barre syndrome, multiple sclerosis (MS) and acute diseminated encephalomyelitis (ADEM) are important in diferential diagnosis. The most promising treatment is plasmapheresis, high dose IV steroid pulse therapy and IVIG therapy. Prognosis is variable and residual symptoms are common. We presented six‐year old boy with acute flaccid paralysis of lower extremity, sensory disturbances and retention of urine. He had prodromal symptoms of vomiting,fever,neckpain and backpain twelve hours before admission. Magnetic resonance imaging of spine on T2 weihgted images ,showed increased signal throught the spinal cord from C7 to Th5 and Th7 to L1 segment.MRI of brain was normal. The cerebrospinal fluid analysis revealed lymphocytic pleocytosis, elevated protein level and normal glucose. On second day child was transported in Zagreb University Hospital for Infectious Diseeases and he was treated with plasmapheresis and high dose IV steroid pulse therapy. After that, he is recovering slowly,step by step; he is still on physical therapy.


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PP96 APPLICATION OF NEUROPHYSIOLOGIC BACKGROUND OF NORMAL MOVEMENT IN PHYSIOTHERAPIST TREATMENT WITH BOBATH CONCEPT BY OPTIMIZING THE FUNCTION OF UPSTANDING ACITIVITIES IN ADULT PATIENTS WITH ACQUIRED LESIONS OF THE CNS ‐ CASE REPORT Jelica Stjepan, Seper Vesna, Davidović Erna Veleučilište Lavoslav Ružička u Vukovaru ‐ studij fizioterapije, Županijska 50, 32000 Vukovar By nonspecific medical gymnastics, patient after insults are achieving certain results in terms of efficiency but not in terms of quality of movement. This is often done through the improvement of the strategy which in patients everyday functioning is causing secondary problems such as pain,additional efforts and motion restrictions. A complex move such is activity of upstanding,consists of several components which are realized throughout certain structures.The role of peripheral control is to ensure that a patient receives the information needed for movement.On the other side, this provides an impact for the hierarchical control and management of movement for a physiotherapist with facilitation and inhibition techniques. This will be done throughout three different forms of physiotherapist's sensomotoric inputs such is handling, verbal facilitation and visual facilitation. The goal of treatment by Bobath concept is development of the function(effectiveness) and optimization of functi on(efficiency ). Bobath concept is today called as neurodeveloping concept which uses the scientifically established facts and neurophysiology as the basis for the treatment of patients. Brief description of the case: Patient at the age of 62 years experienced a stroke.Cured on the department for neurology for 11 days. Specific physiotherapist's treatment was not conducted. Main goals: It is established that the patient have difficulties to get up from the bed(only possible after several attempts). While attempting, the patient uses a strategies and also is feeling a great fear of falling. Main goal was to abandon the strategies and establish selective use of components in the activity of upstanding and to reduce the risk of falling. Method: Work with a patient has been conducted during 25 treatments by Bobath concept (lhour per treatment) A status form of the Bobath concept has been used for the taking of status in accordance with the ICF classification. The quality of the activities for the movement used in upstanding process is analyzed through the occurrence or absence of components of complex activities as well as the quality of them. PP97 TREMOR, SEIZURES, PSYCHOSIS AND DIPLOPIAE IN LYME‐NEUROBORRELIOSIS: REPORTS OF TWO CASES Šarac H1, Markeljević J2 1Croatian Institute for Brain Research, Diagnostic Centre "Neuron", Neurology Department, School of Medicine, University of Zagreb, 2Department of Internal Medicine, University Hospital Centre Zagreb, School of Medicine, University of Zagreb Background: Borrelia burgdorferi (Bb) can cause a large number of neurological symptoms. Although extrapyramidal disturbances, seisures and psychosis are rare, tremor Case report: We described two patients with unusual presentation of Lyme‐neuroborreliosis. An 45‐year old man developed tremor, seizures and psychotic epizodes. Second patient is an 55‐year old man presenting with diplopiae, optic neuropathy and hypoacusis. Standardized medical interview, neurologic examination, neuroimaging, serum and CSF serology and electroencephalographic evaluation have been performed. Both patients were treated with intravenous ceftriaxone and responded with rapid clinical and functional improvement. Anyway, both patients had sequelae that influenced their daily activities post treatment. Emphasis is placed on the atypical onset and evolution, the difficulties encountered in formulating diagnosis, and the uncertainties concerning the sequelae after treatment.


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Conclusion: Early diagnosis and treatment would seen to be of great importance in order to avoid sequelae. In patients with non‐specific long.lasting symptoms in the absence of the overt clinical signs suggesting CNS involvement, routine treatment with i.v. ceftriaxone is not to be encouraged. PP98 DURAL, JUVENILE AND PERIMEDULLARY ARTERIOVENOUS SHUNTS: REPORT OF FOUR CASES Šarac H1, Telarović S2, Vranješ D2, Žagar M2, Markeljević J3, Ozretić D4

1Croatian Institute for Brain Research, Diagnostic Centre "Neuron", Neurology Department, School of Medicine, University of Zagreb, 2Department of Neurology, University Hospital Centre Zagreb, Zagreb School of Medicine, Croatia, 3Department of Immunology and Rheumatology, University Hospital Centre Zagreb, Zagreb School of Medicine, 4Department of Radiology, University Hospital Centre Zagreb, Zagreb School of Medicine Background: Spinal arteriovenous malformations are rare conditions that usually cause subacute necrotizing myelopathy (Foix‐Alajouanine syndrome) or haemorrhagia. Case description: We described four patients with spinal arteriovenous malformations (AVM) who complained back pain, weakness of the lower limbs and sphyncter dysfunction. Thoracolumbar MRI typically showed focal areas of hyperintensity in T2 weighted images and enlarged venous plexus in all cases presented. Spinal angiography revealed three different types of AVM. In the first patient, a 57‐year old man spinal angiography showed dural arteriovenous fistula (AVF). Juvenile perimedullary AVMs has been revealed in the second case of 32‐year old man and juvenile intramedullary AVM in the third case of the 17‐year old girl. Endovascular embolisation resulted in complete closure of the feeding artery in the first patient with dural AV fistula and in the third patient with intramedullary AVM. A good restoration of a new hemodynamic equilibrium between malformation and the cord itself has been reached. Embolisation has not been performed in the second patient with perimedullary AVM, who had been repetitivelly embolized during the past 18 years. Recently, he rapidly deteriorated up to paraplegia during angiography procedure. Embolization also could not been performed in the fourth case of the 59‐year old woman, due to a unremarkable presentation of the multiple feeding arteries of pial AVM. Conclusion: These findings rise our attention about need to keep in mind the clinical suspicion of AVM in cases of back pain, weakness of lower lims and sphincter disturbances. Magnetic resonance imaging (MRI) and spinal angiography of spinal AVM are fundamental methods that may help in definig the real incidence, clinical correlations, and the risk of ishemic and haemorrhagic complications of these vascular malformations. Since ensuing the success of therapy depends on rapid verification of the extent of the illness, early neuroimaging and endovascular embolisation conduct are necessary to prevent paraplegia. Key words: spinal arteriovenous malformation, dural AVF, juvenile AVM, perimedullary AVF. PP99 AGGRESSIVE INTESTINAL SCHWANNOMA MALIGNUM MIMICKING GYNECOLOGICAL PATHOLOGY Tomica D1, Danolić D1, Puljiz M1, Alvir I1, Mamić I1, Milas I2, Knežević F3, Banovic M4 1Department of Gynecologic Oncology, University Hospital for Tumors, Zagreb, Croatia, 2Department of Surgery, University Hospital for Tumors, Zagreb, Croatia, 3Department of Pathology, University Hospital for Tumors, Zagreb, Croatia, 4Department of Transfusion Medicine, University Hospital for Tumors, Zagreb, Croatia Primary malignant schwannoma of the small and large intestine is an extremely rare disease. Therefore we are going to report a aggressive multifocal malignat intestinal schwannoma in 66‐year old female patient, firstly diagnosed as a gynecological tumor, that, even after the surgical treatment, had very quickly recurrence. Tumors were completely removed, but one month after the operation on a check up examination a recurrent tumors were diagnosed. The patient did not suffer from Neurofibromatosis type 1 (NF‐1), disease that increases causality of malignant schwannoma in compare with general population. Diagnosis of these tumors is often late, while radical surgical intervention does not guarantee longer survival. Small intestine tumors may show images similar to an adnexal tumor, so it is difficult to differ them prior to the surgery. Therefore, in presence of a


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pelvic mass, especially if other unusual amnestic data are present, the possibility of other than a gynecologic tumor has to be considered. Key words: malignant, aggressive, schwannoma, gynecologic


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