Official Journal of the Romanian Society of Physiological Sciences

CHIEF EDITOR
CO-CHIEF EDITORSFRANCISC SCHNEIDERIOANA SISKACARMEN TATUASSOCIATE EDITORSMIHAI NECHIFORSORIN RIGAEXECUTIVE EDITORSFLORINA bojin
GABRIELA TANASIE
daciana nistor
	cALIN MUNTEANEDITORIAL BOARD
ARDELEAN AURELBADIU GHEORGHEBĂDĂRĂU ANCA
BENEDEK GYÖRGYBENGA GHEORGHEBUNU CARMEN
COJOCARU MANOLECUPARENCU BARBUCONSTANTIN NICOLAE
HAULICĂ ION
IANCĂU MARIAMIHALAŞ GEORGETA
MUNTEAN DANINAMUREŞAN ADRIANANESTIANU VALERIUOPREA TUDOR(Arad)(Constanţa)(Bucureşti)(Szeged)(Cluj)(Timişoara)(Bucureşti)(Oradea)(Bucureşti)(Iaşi)(Craiova)(Timişoara)(Timişoara)(Cluj)(Craiova)(New Mexico)PĂUNESCU VIRGILPETROIU ANA
POPESCU LAURENŢIU
RÁCZ OLIVERRIGA DANSABĂU MARIUS
SIMIONESCU MAIA
SIMON ZENOSAULEA I. AURELSWYNGHEDAUW BERNARDTANGUAY M. ROBERTTATU FABIAN ROMULUS
VLAD AURELIANVOICU VICTORZĂGREAN LEON

(Timişoara)(Timişoara)(Bucureşti)(Košice)(Bucureşti)(Tg. Mureş)(Bucureşti)(Timişoara)(Chişinău)(Paris)(Canada)(Timişoara)(Timişoara)(Bucureşti)(Bucureşti)Accredited by CNCSIS - B category - code 240
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Fiziologia ‑ Physiology  2011 supplement

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Official Journal of the Romanian Society of Physiological Sciences

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Fiziologia ‑ Physiology  2011 supplement

“Doua tari, un scop, succes comun!”
Project acronym: HU-RO DOCS
Project title: Hungarian-Romanian multidisciplinary PhD doctoral school programme in medicine
Reg. no: HURO/0901/069/2.3.1

Bilateral Cooperation of Doctoral Schools Timisoara-Szeged –an European Model for Setting up a Doctoral Programme in the Medical Field
University of Medicine and Pharmacy Victor Babes Timisoara,
Piata Eftimie Murgu, No. 2, Room 215, 2nd floor

Conference Programme, 05-06.05.2011
THURSDAY, 05.05.2011
14.00-14.15

Opening ceremony

PAPER SESSION I
14.15-14.35
14.35-14.55
14.55-15.10
15.10-15.25
15.25-15.40
15.40-15.55
15.55-16.10
16.10-16.40

Prof. Dr. Adrian Neagu: Biological Physics Tools for Investigating Multicellular Systems
Prof. Dr. Ferenc Bari: Optical Methods for Studying Cerebro-Cortical Microcirculation
PhD Stud. Şerban Comşa: Correlations between the Vascular Endothelial Growth Factor
Expression, Microvascular Density in Tumor Tissues and the TNM Staging in Breast Cancer
PhD Stud. Dénes Garab: Beneficial Effects of Remote Limb Ischemic Preconditioning
against the Microcirculatory Consequences of Partial Hepatic Ischaemia in Rats
PhD Stud. Ştefana Feflea: Evaluation of EG-VEGF Antibody Effect on Chick Embryo
Chorioallantoic Membrane and Organs
PhD Stud. László Juhász: Examination of the Trigger Role of Peroxynitrite in the
Antiarrhythmic Effect of Ischaemic Preconditioning and Peroxynitrite Infusion
PhD Stud. Adina Duşe: Expression and Possible Significance of Vascular Endothelial Growth
Factor in Non Hodgkin Lymphoma
Coffee Break

PAPER SESSION II
16.40-17.00
17.00-17.15
17.15-17.30
17.30-17.45
17.45-18.00
18.00-18.15
19.00

Prof. Dr. Carmen Bunu Panaitescu: Dendritic Cells Response to in vitro Allergen Challenge
and Impact of Corticosteroids on Local Inflammation
PhD Stud. Adrian Gruici: Carotid Intima-Media Thickness and the Prediction of Clinical
Cardiovascular Events in Patients with Manifest Arterial Disease
PhD Stud. Tünde Tőkés: Protective Effect of L-a Glycerylphosphorylcholine against
Ischaemia-Reperfusion Injury in the Rat Small Intestine
PhD Stud. Laura Marusciac: Specific Chemokine-Chemokine Receptors Interactions andTheir Role in hMSC Migration
PhD Stud. Gabor Kisvári: Acute Administration of Simvastatin Reduces the Severity ofVentricular Arrhythmias in Anesthetized Dogs
PhD Stud. Adelina Mavrea: Ventricular Asynchronysm - Prognosis Marker in Heart Failure
with Preserve Ejection Fraction
Dinner

FRIDAY, 06.05.2011
PAPER SESSION III
9.00-9.20
9.20-9.35
9.35-9.50
9.50-10.05
10.05-10.20
10.20-10.35
10.35-11.05

Prof. Dr. Mihály Boros: Ischemia-Reperfusion Injuries, from Laboratory Studies to
Clinical PracticePhD Stud. Simona Bota: The Value of Acoustic Radiation Force Impulse Elastography (ARFI)
for the Assessment of Chronic Hepatopathies
PhD Stud. Csaba Gombár: Effect of Calcaneo-Stop Procedure on Load Conditions ofPaediatric Flexible Flatfoot
PhD Stud. Bogdan Bălinişteanu: Prevalence of Ki-67 in Pitutary Adenomas
PhD Stud. Bernadett Borda: Functional and Histopathological Changes in Renal TransplantPatients with New-Onset Diabetes and Dyslipidemia
PhD Stud. Viorica Bocan: Neuroendocrine Differentiation in Benign Prostate Diseases. An
Autopsy Report
Coffee Break

PAPER SESSION IV
11.05-11.25
11.25-11.40
11.40-11.55
11.55-12.10
12.10-12.25

12.25-12.40
12.40-14.00

Assoc. Prof. Dr. Gyula Sáry: Beyond the Orbit - Cortical Mechanisms of Shape and Form
VisionPhD Stud. Nóra Szabó: Epidemiology of Central Nervous System Malformations in a Regionin Hungary
PhD Stud. Diana Naiche: Mesostructure. Types of Bars in Implant Overdenture
PhD Stud. Levente Szalardy: Study on the Levels of Tau and b-amyloid in CSF of Patientswith Multiple Sclerosis
PhD Stud Claudia Corici: The Properties of the Transient Outward, Inward Rectifier andAcetylcholine Sensitive Potassium Currents in Atrial Myocytes from Dogs in Sinus Rhythm
and Experimentally Induced Permanent Atrial Fibrillation
PhD Stud. Magdolna Kósa: Efficiency of Plasmid Transfection along the Regenerating Rat
Soleus Muscle
Lunch

PAPER SESSION V
14.00-14.20
14.20-14.35
14.35-14.50
14.50-15.05
15.05-15.20
15.20-15.35
15.35-15.50
15.50-16.05
16.05-16.15

Assoc. Prof. Dr. Cristina Dehelean: New Trends in Toxicological Screening Applied onExperimental Models Correlated to Tumours
PhD Stud. Péter Szerémy: Investigation of Transporter Interactions of Antimalarials In Vitro
PhD Stud. Tuboly Eszter: The Effects of Exogenous Methane Inhalation on Macro- andMicrocirculatory Changes during Intestinal Ischemia-Reperfusion in Rats
PhD Stud. Vanya Melinda: Childbearing among Women with Multiple Sclerosis (MS)PhD Stud. Simona Trancotă: Characterization of the Effects of Two Porphyrin Compounds
on Rat Heart Mitochondrial Respiration
PhD Stud. Bere Zsófia: Multimodal, Live Imaging of Global Forebrain Ischemia- Induced
Periinfarct-Depolarisation (PID) in the Rat Cerebral Cortex
PhD Stud. Georgel Ţăranu: Ischemic Limb Salvage Using Neoangiogenic Therapy – Case
ReportPhD Stud. Emese Asztalos
Concluding Remarks

Official Journal of the Romanian Society of Physiological Sciences

1. PREVALENCE OF KI-67 IN PITUTARY ADENOMAS
Bogdan Balinisteanu, Raluca Ceausu, Anca Maria Cimpean,
Marius Raica
Victor Babes University of Medicine and Pharmacy Timisoara,
Department of Histology and Molecular Pathology, Angiogenesis
Research Center Timisoara
Tutor: Prof. Dr. Marius Raica, Victor Babes University of Medicine
and Pharmacy Timisoara
E-mail: med_bogdan@yahoo.com
Introduction. Pituitary adenomas represent the third mostcommon primary intracranial tumors in neurosurgical practice.
To understand the biological behaviour of the pituitary adenomas,
cell proliferation markers, such as monoclonal antibodies targetedagainst the Ki-67 antigen, have been applied. The Ki-67 antigen isa protein related to cell proliferation and is expressed in cell nuclei
throughout the entire cell cycle. Aim. The aim of this study was tocorrelate the Ki-67 index with hormone phenotype of pituitaryadenomas. Materials and methods. Our study included 30 casesof pituitary adenomas. Sections from each case were stainedwith routine haematoxylin and eosin method for histopathologic
evaluation. Immunohistochemistry was performed on additionalslides in order to detect specific pituitary adenomas. The percentage
of Ki-67–positive nuclei (the Ki-67 Label Index) was determinedby counting approximately 1000 nuclei of tumor cells at 400-foldmagnification. Results. Ki-67 immunostaining was detected in 21
cases, with values between 0.2% and 4.8%.The highest values were
noticed in 3 prolactin (PRL) -secreting adenomas (range 3.1 – 4.8%).
The Ki-67 was negative in 9 cases, the majority of them being nonsecretory adenomas (6 of 9). Discussion. Pituitary adenomas areheterogeneous in growth rate, invasiveness and recurrence. Theyfrequently invade surrounding structures such as the cavernous sinus,
sphenoid sinus, and even the brain. Conclusions. The Ki-67 index
was significantly higher in adenomas with prolactin secretion.
Keywords: Immunohistochemistry, Ki-67, pituitaryadenomas

2. MULTIMODAL, LIVE IMAGING OF GLOBALFOREBRAIN ISCHEMIA- INDUCED PERIINFARCTDEPOLARISATION (PID) IN THE RAT CEREBRALCORTEX
Zsófia Berea,b, Adrienn Kulmánya,b, Tihomir P. Obrenovitcha,c,Ferenc Baria, Eszter Farkasa,bDepartment of Medical Physics and Informatics, Faculty of
Medicine, University of Szegedb
Department of Physiology, Faculty of Medicine, University ofSzegedc
Division of Pharmacology, School of Life Sciences, Universityof Bradford, United KingdomaTutor: Prof. Dr. Ferenc Bari, University of Szeged
E-mail: berezsofia@gmail.com
Propagating electrical inactivation called periinfarct-depolarization(PID) and associated changes in cerebral blood flow (CBF) spontaneously
occur in the cortex during cerebral ischemia. These phenomena

Fiziologia ‑ Physiology  2011 supplement

may contribute to the extension of brain infarcts. We have set out
to characterize PID-related changes in membrane potential andhemodynamic variables in rat cerebral cortex.
In anesthetized, male Sprague-Dawley rats (n=13) a closed
cranial window was mounted over the parietal bone and loadedwith voltage sensitive (VS) dye. The left femoral artery and vein
were cannulated for the monitoring of mean arterial pressure(MAP) and for blood withdrawal. Global forebrain ischemia was
induced by bilateral common carotid artery occlusion combinedwith subsequent hypovolemic hypotension (MAP<40 mmHg).
Using selected illuminations we acquired synchronous changes inmembrane potential (VS dye method); cerebral blood volume withgreen (540–550 nm); hemoglobin saturation with red (620–640nm) illumination, and CBF by laser speckle contrast imaging.
PID occurred at a MAP value of 41.2±3.7 mmHg and at a CBF
value of 43.4±4.9%. In most cases the depolarization was notfollowed by the recovery of membrane potential, and these eventswere associated with CBF reduction (19±3.6%).
In conclusion, PID appears to generate at the lower limit of
the autoregulatory range of CBF, originates at a focus with highvulnerability to ischemia, and propagates similar to that known
for cortical spreading depression. PID associated CBF responses
most often display inverse neurovascular coupling.
This work was supported by grants from the EGT NorwegianFinancial Mechanisms (NFM, NNF 78902), and Hungarian Scientific
Research Fund (OTKA, K81266).
Keywords: multimodal imaging, global forebrain ischemia,
periinfarct-depolarisation, neurovascular coupling

3. NEUROENDOCRINE DIFFERENTIATION IN BENIGNPROSTATE DISEASES. AN AUTOPSY REPORTElena Viorica Bocan1, Horatiu Ieremia2, Dorin Agapie3,
Ovidiu Mederle1, Radu Minciu4, Marius Raica1
Victor Babes University of Medicine and Pharmacy Timisoara,
Cytology and Histology Department
2
Clinical Hospital Zalau
3
Clinical Hospital Timisoara, Oncology Department
4
Judetean Hospital Timisoara, Urology Department

1

Tutor: Prof. Dr. Marius Raica, 1Victor Babes University of Medicine
and Pharmacy Timisoara
E-mail: viobocan@gmail.com
Basal cell hyperplasia is occasionally a component of untreatedusual, nodular, glandular and stromal hyperplasia. Prostatic
intraepithelial neoplasia (PIN) has been identified as precursor
lesions to prostatic carcinoma.
The aim of the study was to identify and evaluate neuroendocrine
differentiation in autopsy cases of patient that died of other causes
than prostate lesions.
The study was performed on a number of 17 cases. The cases havebeen harvested from the Forensic Medical Institute from patients
without prostatic pathology. 3µm sections were performed andthen immunohistochemical marker antibodies anti-chromogranin
A and PSCA (prostate stem specific antibody) were applied.
After standard hematoxylin-eosin staining was performed, a seriesof benign prostatic lesions were identified: basal cell hyperplasia,
prostatic intraepithelial neoplasia, cystic and lobular degeneration.
In some sections, zones of post-mortem autolysis of various sizes

5Official Journal of the Romanian Society of Physiological Scienceswere identified. The presence of neuroendocrine differentiation wasproved in both small normal prostate zones and also at the level ofbenign lesions. PSCA was negative for neuroendocrine cells. Asideof neuroendocrine differentiation, neuroendocrine cell hyperplasia
was identified (more than 3 cells per gland) on the same type oflesions. The neuroendocrine differentiation was identified on anumber of 10 cases out of the 17 cases. In the same sections we
identified hyperplasia of neuroendocrine cells.
PSCA has been shown as negative marker of neuroendocrine
cells with positive control in secretory cells. We found neuroendocrine
differentiation in cases of patients without prostate pathologicalconditions, diagnosed post–mortem with benign prostate lesions.
Keywords: post mortem diagnosis, benign lesion, chromogranin
A, neuroendocrine differentiation

4. FUNCTIONAL AND HISTOPATHOLOGICALCHANGES IN RENAL TRANSPLANT PATIENTS WITH
NEW-ONSET DIABETES AND DYSLIPIDEMIA
Bernadett Borda, György Lázár
University of Szeged, Department of Surgery, Szeged, Hungary
Tutor: Prof. Dr. György Lázár, University of Szeged
E-mail: bb@surg.szote.u-szeged.hu
The principal risk factors of post-transplantation cardiovascularmortality are hyperglycemia, hypertriglyceridemia, obesity andsmoking.
115 patients were included in the study. We assessed the risk
factors of diabetes (NODM) and dyslipidemia (NODL), and theeffect of these on the function and histopathological changes of
the allograft, one year after the transplantation.
When evaluating the risk factors and the initial recipient data,
we found a significant difference in age when comparing normal (N)and NODM patients (p=0.004), N and NODL patients (p=0.002) and N
and NODL+NODM patients (p=0.0001). The difference in body massindex (BMI) was significant when comparing N and NODM+NODL
patients (p=0.003). In regard to immunosuppressive therapy, NODM
was significantly more frequent in those taking tacrolimus (Tac)(p=0.005), whereas those who received cyclosporine (CsA) showeda significantly higher incidence of NODL (p=0.001). The triglyceride
level was 3.02±1.51 mmol/L in those taking CsA and 2.15±1.57 mmol/Lin those taking Tac, which is a significant difference (p=0.004). The
difference proved to be significant also when assessing the totalcholesterol level: it was 5.43±1.23 mmol/L in those receiving CsAbased immunosuppression and 4.42±1.31 mmol/L in patientstaking Tac (p=0.001). In regard to allograft function, one year after
the transplantation a significant difference was found between the
NODM+NODL group and the N group in serum creatinine level(p=0.02), as well as in eGFR (p=0.004).
When assessing the morphological changes of the kidney, we
found that interstitial fibrosis/tubular atrophy occurred significantly
more frequently in all three groups, compared to normal patients.
Our own clinical study proves that one year after transplantation
the allograft function is already impaired if both medical conditions(NODM and NODL) are present; however, in regard to morphology, asingle condition (NODM or NODL) is sufficient to lead to histologicalchanges in the kidney.
Keywords: new-onset diabetes mellitus and dyslipidemia

6

5. The value of Acoustic Radiation Force
Impulse Elastography (ARFI) for the assessment of chronic hepatopathies
Simona Bota, I. Sporea, Roxana Şirli, Alina Popescu,
Mirela Dănilă
Department of Gastroenterology and Hepatology, Victor Babes
University of Medicine and Pharmacy Timisoara
Tutor: Prof. Dr. Ioan Sporea, Victor Babes University of Medicine
and Pharmacy Timisoara
E-mail:: bota_simona1982@yahoo.com
Introduction: ARFI is a new noninvasive method for theassessment of liver fibrosis.
Aim: To assess the value of liver stiffness measurement bymeans of ARFI as a predictive factor for the severity of fibrosis andfor the presence of cirrhosis complications.
Methods: We performed ARFI measurements in 523 patients(p),mean age 59.6±10.2 years, 210 women and 313 men. In 150p
(28.6%) we performed liver biopsy and 373p were diagnosed withliver cirrhosis by clinical ultrasound, endoscopy criteria. In 163
cirrhotic patients we permormed also ARFI in spleen. 10 valid ARFI
measurements were performed in each patient and a median value
was calculated, expressed in meters/second (m/s).
Results: We obtained valid liver ARFI measurements in 511p
(97.7%). In patients with liver biopsy we obtained a direct, liniar
correlation between ARFI values (r=0.527) and liver fibrosis (p<0.0001).
The correlation was stronger in patients with HCV vs. HBV
hepatitis, but not statistically significant (r=0.557 vs. r=0.49,
p=0.67). The medium ARFI value in cirrhotic patients with significant
esophageal varices-EV (al least grade II-150p) was not statisticallysignificant vs. those without or with grade I EV (157p): 2.8±0.73 vs.
2.93±0.72m/s (p=0.13), also between patients with (69p) or without
hepatocellular carcinoma (287p): 2.76±0.73 vs. 2.86±0.72m/s(p=0.27). ARFI values were statistically higher in patients with ascites(140p) vs. those without ascites (216p):2.99±0.71 vs. 2.74±0.71m/s(p=0.003). Spleen ARFI values were statistically higher in patientswith significant EV (95p) vs. those without EV or grad I EV (68):3.31±0.50 vs. 3.09±0.56, p=0.01. For a cut-off value >3.47 m/s , ARFIin spleen had 43.6%Se, 80.3%Sp, 75.9%PPV, 50%NPV, 58.7% accuracy(AUROC=0.60) for the prediction of significant EV.
Conclusion: ARFI is a good method for the assessment of liver
fibrosis. In cirrhotic patients can predict the vascular decompensation
and ARFI values in the spleen can predict the significant EV.
Key words: liver fibrosis, Acoustic Radiation Force Impulse
Elastography (ARFI), liver stiffness, spleen stiffness, liver cirrhosis

6. CORRELATIONS BETWEEN THE VASCULAR
ENDOTHELIAL GROWTH FACTOR EXPRESSION,MICROVASCULAR DENSITY IN TUMOR TISSUESAND THE TNM STAGING IN BREAST CANCER
Serban Comsa, Anca Maria Cimpean, Raluca Ceauşu,Cristian Suciu, Marius Raica
Victor Babes University of Medicine and Pharmacy Timisoara,
Department of Histology
Tutor: Prof. Dr. Marius Raica, Victor Babes University of Medicine
and Pharmacy Timisoara

Fiziologia ‑ Physiology  2011 supplement

Official Journal of the Romanian Society of Physiological Sciences
E-mail: serbancomsa@yahoo.com
Background: The breast cancer is the most frequent malignancy
of women all over the world. In spite of its doubtable reliability, theTNM staging system is widely used for the prognosis of this disease.
On the other hand, angiogenesis is considered to have an essential
role in the evolution of breast cancer and the Vascular Endothelial
Growth Factor (VEGF) has proven to be the key regulator of thisprocess. Quantitation of VEGF and the tumor vasculature mightplay an important role in predicting tumor behavior and patientmanagement. The aim of our study was to evaluate the potential
correlation between the VEGF expression, microvessel density (MVD)
in the tumor tissues and the TNM staging in breast cancer.
Method: We included 34 pacients with breast cancer, who
had undergone surgical treatment, and we evaluated each casehistopathologically and by immunohistochemistry for VEGF and
CD31 expression in the tumor tissues. VEGF expression was evaluatedby calculating the average percentage of cytoplasmic positive cells
from 3 high power fields. MVD was expressed as the average numberof the CD31+ microvessels from 3 high power fields.
Results: Expression of VEGF was significantly associatedwith the number of lymph-nodes with metastasis, the number ofcells in mitosis, the presence of necrosis and the T-stage (inverse
correlation). MVD correlated very well with the histological grading,the number of cells in mitosis, the presence of inflamation and thepresence of necrosis.
Conclusion: Although their relationship with the TNM staging
remains unclear, VEGF expression and MVD prove to be important
indicators of the malignant status in breast cancer, confirming the
major involvement of angiogenesis in this type of cancer. Both ofthem are valuable prognostic factors in breast cancer, but the patternof their relationship needs further analysis of the VEGF receptors
in order to be described.
Keywords: angiogenesis, breast cancer, microvessel density,
TNM staging, VEGF

7. The properties of the transient outward, inward rectifier and acetylcholine
sensitive potassium currents in atrial
myocytes from dogs in sinus rhythm
and experimentally induced permanent
atrial fibrillation
Claudia Corici1, Zsófia Kohajda2, Attila Kristóf 2, László
Virág1, Zoltán Husti1, Viktor Juhász1, Chadaide Szami3,
Sághy László3, István Baczkó1, András Varró1,2, Norbert
Jost1,2
Department of Pharmacology & Pharmacotherapy, Universityof Szeged
2
Division of Cardiovascular Pharmacology, Hungarian Academyof Sciences, Szeged
3
2nd Department of Medicine and Cardiology Center, Universityof Szeged1

Tutor: Assoc. Prof. Dr. Norbert Jost, University of Szeged
E-mail: claudy_corici@yahoo.com
Background: The aim of present study is to investigate andcompare the properties of repolarizing currents which contributeto AF induced remodeling, i.e. the transient outward (Ito), inward

Fiziologia ‑ Physiology  2011 supplement

rectifier (IK1), acetylcholine sensitive potassium currents (IK,ACh) in
isolated atrial canine myocytes obtained from normal (SR) andtachypaced atrial fibrillating (ATR) dogs by applying in vitro (patch
clamp) and in vivo electrophysiological techniques.
Results: Ito current was slightly downregulated in ATR cells
when compared with that of recorded in SR. IK1 was larger in the
myocytes isolated from ATR dogs, however, this upregulation is
not so extensive between -80-20 mV, the voltage range close to
an atrial action potential (AP). IK,ACh was activated by cholinerg
agonist carbachol (2 µM). In SR, carbachol activated a large
current either at outward or inward directions. Selective IK,AChblocker, tertiapin (10nM) blocked the carbachol induced current
by 57%. In atrial myocytes from ATR dogs we could measure thepresence of a constitutively active IK,ACh, which could be blocked
by 26% with 10nM tertiapin. However, in atrial myocytes from
ATR dogs, carbachol in addition could also activate a significant
ligand-dependent and tertiapin sensitive IK,ACh current. In vivo
application of tertiapin in ATR dogs even in low concentration(<3 nM) effectively prevented burst induced AF.
Conclusion: In contrast to the human, ATR slightly downregulated
I to in dogs. The slower I to current inactivation may contribute
more to atrial repolarization than that of described in ventricular
myocytes. The presence of the constitutively activated IK,ACh in
atrial myocytes from ATR dogs shows that electrical remodelingdeveloped in our model. The constitutively active and liganddependent IK,ACh currents together play an important role in dogs
by contributing to AF remodeling, since selective IK,ACh blockade
effectively prevented AF.
Keywords: atrial fibrillation (AF), tachypaced atrial fibrillating(ATR) dogs, AF induced remodeling

8. EXPRESSION AND POSSIBLE SIGNIFICANCEOF VASCULAR ENDOTHELIAL GROWTH FACTORIN NON HODGKIN LYMPHOMADuşe Adina Octavia1, Tibor Mezei2, Amalia Raluca Ceauşu1,
Anca Maria Cimpean1, Marius Raica1, Hortensia Ionita3
1-Department of Histology, Victor Babeş University of Medicine
and Pharmacy Timişoara
2-Department of Pathology, University of Medicine and Pharmacy Tîrgu Mureş
3- Department of Hematology, Victor Babes University of Medicine and Pharmacy Timisoara
Tutors: Prof. Dr. Hortensia Ionita; Prof. Dr. Marius Raica, Victor
Babes University of Medicine and Pharmacy Timisoara
E-mail: adinadusche@yahoo.com
Background. Non-Hodgkin’s lymphoma (NHL) is a heterogenous
group of lymphoid malignancies, characterized by unpredictablebehaviour. The potential significance of angiogenesis in NHL isrelated to the association between the disease progression andthe increased angiogenic activity. VEGF plays a crucial role in bothnormal and tumor-associated angiogenesis and together with its
receptors, it has been shown to represent a promising target for
antiangiogenic therapy. Lymphoma cells express VEGFR-1 and
VEGFR-2 which promote survival, proliferation and metastasis
through autocrine mechanisms. Only few things are known about
the exact trigger mechanisms of VEGF expression in hematolymphoid malignancies, but the recent findings have showed7Official Journal of the Romanian Society of Physiological Sciences
some similar features to those observed in solid tumors.
The aim of this study was to investigate the expressionof VEGF in malignant non-Hodgkin lymphoma, in terms of its
immunohistochemical distribution and clinico-pathologicalsignificance.
Material and methods. Twenty one specimens of lymph nodebiopsies were included in this study. For the imunohistochemical
procedure we used anti-VEGF antibody clone VG-1 (ready-to-use,
30 minutes incubation time at RT), avidin/biotin/HRP based working
system, and 3,3 diaminobenzidine as chromogen. Counterstain
was performed by using modified Lille’s hematoxylin. We evaluatedthe distribution in tumour cells, macrophages and in nontumoral
cells. VEGF immunoreactivity was estimated as percent of positive
cells according to this score: 0 (0% positive cells); 1 (<10%); 2(10-30%); 3 (>30%).
Results. Histopathologic evaluation revealed 18 cases ofdiffuse type lymphoma and 3 cases of follicular type lymphoma.
VEGF was pozitive in 95,23% of cases and in one case, VEGF was
negative in all cell types. We noticed a significant correlation in
VEGF expression between tumor cells and macrophages (p=0,001).
It was also found a significant correlation between tumor cells
and nontumoral cells (p=0,002).
Conclusion. In non-Hodgkin lymphoma the main mechanismof angiogenesis seems to be dependent on the VEGF pathway
and its expression particularly by stromal cells.
Key words: angiogenesis, lymph node, imunochemistry,
VEGF

9. EVALUATION OF EG-VEGF ANTIBODY EFFECTON CHICK EMBRYO CHORIOALLANTOIC MEMBRANE AND ORGANS
Stefana Feflea1, Raluca Amalia Ceausu2, Pusa Gaje2, Anca
Maria Cimpean2, Marius Raica2
Victor Babeş University of Medicine and Pharmacy Timisoara,
Department of Pharmacognosy and Department of Histology,
Angiogenesis Research Center Timişoara
2
Victor Babeş University of Medicine and Pharmacy Timisoara,
Department of Histology, Angiogenesis Research CenterTimişoara
1

Tutor: Prof. Dr. Marius Raica, Victor Babeş University of Medicine
and Pharmacy Timisoara
E-mail: stefana.feflea@gmail.com
Introduction. Endocrine gland derived vascular endothelial
growth factor (EG-VEGF), also called prokineticin 1 (PK1) is a highly
specific mitogen that regulates proliferation and differentiationof the vascular endothelium in a tissue specific manner. The
inhibition of EG-VEGF/ PK receptor pathway could represent a
more selective antiangiogenic and anticancer strategy. This study
aimed to evaluate the modifications produced by an antibody anti
– EG-VEGF on the vascularized chick chorioallantoic membrane(CAM) and in embryonic organs.
Material and methods. The in vivo CAM assay was performedin ovo, by injecting different dilutions of the antibody EG-VEGF(T-16) in the allantoic vesicle of the embryo, from incubation
day 7. Test and control specimens were macroscopic observed,
histologically processed and interpreted.
Results. Hemorrhagic areas of the capillary plexus were8

observed after only one day of administration of the antibody
which can be correlated with a very low survival rate. On
hematoxylin-eosin staining of the CAM, there was evidenceof vascular disruption and erythrocyte extravasations in thechorion. There were signs of CAM epithelium vacuolization
while the high density stroma showed a tendency for bloodvessel dilatation and bundle-like organization of medium
and large vessels. The organs of the treated specimens werealso investigated, indicating massive hemorrhagic areas
and blood stasis in brain and lungs. These signs of vascular
impairment could be possibly explained by the role of the
EG-VEGF in the vascular and tissue development of embryonicmembranes, eventually blocked by the specific inhibition of
EG-VEGF; nevertheless different signaling pathways seem to
be involved.
Conclusion. An antibody targeting the endogenous EG-VEGF
might represent a possible anticancer strategy involving pathwaysof impairing angiogenesis, mesenchymal and epithelial structure
in endocrine - derived embryonic tissues.
Keywords: EG-VEGF, angiogenesis, chick chorioallantoic
membrane

10. BENEFICIAL EFFECTS OF REMOTE LIMBISCHEMIC PRECONDITIONING AGAINST THE MICROCIRCULATORY CONSEQUENCES OF PARTIAL
HEPATIC ISCHAEMIA IN RATS
Dénes Garab1, Petra Hartmann1, Rene H. Tolba2, Mihály
Boros1 and Andrea Szabó1,
University of Szeged, Institute of Surgical Research, Szeged,
Hungary2
RWTH Aachen University, Institute of Laboratory Animal
Science and Experimental Surgery, Aachen, Germany

1

Tutors: Dr. Andrea Szabó; Prof. Dr. Mihály Boros, Universityof Szeged
E-mail: humandome@gmail.com
Objective: We investigated whether the harmful microcirculatory
consequences of liver ischemia can be modulated by a remoteischemic preconditioning protocol which can easily be performed
and potentially accessible in the clinical practice
Methods: Partial hepatic ischemia (I/R) was elicited in maleSprague-Dawley rats by occluding the portal vessels of the left
liver lobe for 60 min. The microcirculatory changes induced by I/R
were examined by two methods; leukocyte-endothelial interactions
were determined by fluorescence intravital videomicroscopy,
while tissue perfusion and oxygen saturation changes by the O2C(“oxygen-to-see”) technique. The reperfusion-caused alterations
were compared in I/R and sham-operated animals and in rats thatwere also subjected to limb ischemic preconditioning (IPC, 2 x
10 min I/R) prior to hepatic I/R (n=5-8). Hepatic myeloperoxidase(MPO) enzyme activities and serum AST, ALT, LDH levels were
measured.
Results: I/R caused a significant, 4-fold increase in thenumber of rolling leukocytes and a 3-fold increase of adherent
leukocytes in the central venules of the liver as compared to thevalues of sham-operated animals (rolling: 64±12 1/mm/s; sticking:61±7 1/mm2). The blood flow and tissue oxygen saturation were

Fiziologia ‑ Physiology  2011 supplement

Official Journal of the Romanian Society of Physiological Sciences
decreased by approx. 20% (from control values of ~230 AU and~46%, respectively), while MPO activity increased approx. 4-foldin comparison with those in the sham-operated animals. All of theabove parameters were ameliorated significantly by limb IPC.
Conclusions: Remote IPC exerts marked protection against
the detrimental microcirculatory consequences of hepatic
I/R. Preconditioning applied on the extremities may represent
a potential, clinically applicable approach to ameliorate thepostischemic microcirculatory failure. (Supported by OTKA K75161,
TÁMOP-4.2.2-08/1-2008-0013, TAMOP 4.2.1/B-09/1/KONV-20100005 grants
Keywords: microcirculation, partial hepatic ischemia, limb
ischemic preconditioning, myeloperoxidase

11. Effect of calcaneo-stop procedureon load conditions of paediatric flexible flatfootCsaba Gombar, Kalman Toth, Peter Kellermann
University of Szeged, Department of Orthopaedics, Szent-Györgyi
Albert Medical Center, Hungary
Tutor: Prof. Dr. Toth Kalman, University of Szeged
E-mail: csaba.gombar@yahoo.co.uk
Background: Flatfoot is a frequent postural deformity inchildhood and adolescence. Operative methods may be consideredin severe cases, after ineffective conservative treatment, where thecalcaneus is in remarkably valgus alignment and pain is associated.
Aim of this study is to evaluate the pedographic outcome of
percutaneous lateral arthroereisis with use of a screw throughsinus tarsi into the talus.
Materials and methods: 25 patients (44 feet) underwent
calcaneo-stop surgery at our department between 1st June 2008
and 31st January 2010. Mean age was 10 years (7-14, SD 2.04). Solepressures were recorded with the EMED pedographic analyser before
surgery. Of these, 17 cases (28 feet) were available for follow-up.
The mean follow-up time was 9.7 (7-19, SD 4.98) months.
Results: Area, relative contact time and pressure-time integral(load amount) values increased after surgery on lateral heel, midfootand metatarsal sections. On medial areas a decrease of mentioned
parameters was observed. The most remarkable differences were
measured at midfoot region. Here the contact area value mediallydecreased from pre op 19.69 (2-28, SD 6.61) cm2 to post op 7.31
(1-22, SD 5.6) cm2, pressure-time integral increased laterally from3.99 (0.88-7.31, SD 1.48) Ns/cm2 to 8,44 (2.3-19.88, SD 3.56) Ns/cm2
after surgery. Total contact area of the sole decreased post op
from 120.00 (82-176, SD 27.16) cm2 to 107.23 (73-149, SD 20.856)
cm2. These changes are significant (p<0.001).
Conclusion: Calcaneo-stop procedure seems to provide an
excellent correction for calcaneovalgus deformity at severe flexible
paediatric flatfoot. Pedographic analysis of sole after operation
showed a load shift from medial to lateral, predominantly on
the midfoot and hindfoot, giving a load map close to normal.
Further investigation are required to evaluate the long term
outcome of this method, included the conditions following
implant removal.
Keywords: Paediatric flatfoot; Calcaneovalgus; Calcaneo-stop;
Arthroereisis

Fiziologia ‑ Physiology  2011 supplement

12. CAROTID INTIMA-MEDIA THICKNESS AND THE
PREDICTION OF CLINICAL CARDIOVASCULAR EVENTSIN PATIENTS WITH MANIFEST ARTERIAL DISEASE
Adrian Gruici1, Elena Ardeleanu2, Daniela Gurgus2
Family Medicine Office, Timiş County
Family Medicine Department, Victor Babes University of Medicine
and Pharmacy Timisoara

12

Tutor: Prof. Dr. Elena Ardeleanu, Victor Babes Universityof Medicine and Pharmacy Timisoara
E-mail: adriangruici@yahoo.com
Introduction - Carotid intima-media thickness (IMT) is anindependent predictor of cardiovascular events in the general
population and an important marker regarding the evaluation of
subclinical vascular damage. Its use relies on its ability to predict
future clinical cardiovascular end points.
Aims - Currently, little is known about the relationship between
IMT and new cardiovascular events in patients with manifest
arterial disease. We investigated whether IMT is associated withcardiovascular risk in patients who already have vascular diseaseor atherosclerotic risk factors.
Materials and Methods – The study was performed between
2008-2011 and included 283 patients with manifest arterial diseaseor atherosclerotic risk factors, aged >50 years, mean age 63±5 years,
68% men and 32% females, registered in family medicine offices inTimiş County. IMT was measured at baseline in both carotid arteries
by high resolution ultrasound as agreed in Mannheim consensus.
Increased IMT (>0.9 mm) or presence of carotid atherosclerotic
plaques are evidences of subclinical vascular damage. A value of
IMT, measured by echo-Doppler carotid, greater than 0.9 mm shows
vascular lesions earlier than ankle-brachial index.
Results – Major vascular events were vascular death, ischaemic
coronary events, or stroke. Adjusted for age and sex, an increase incarotid IMT of 1 SD (~0.32 mm) was associated with an increased risk ofany vascular event (Hazard Ratio-HR 1.17; 95% CI 1.04-1.30). Increasing
IMT was most strongly related to ischaemic stroke incidence (HR 1.37;
95% CI 1.16-1.58), than to myocardial infarction incidence (HR 1.21;
95% CI, 1.04 to 1.38). Results were similar in patients without large
carotid plaques (IMT<2 mm at all measurements sites).
Conclusions - Carotid IMT is a strong predictor of futurecardiovascular events. IMT is associated with the occurrence of
new vascular events, mostly for ischaemic stroke, in patients withmanifest arterial disease or atherosclerotic risk factors.
Keywords: Intima-media thickness, cardiovascular events,
atherosclerotic risk factors, stroke.

13. EXAMINATION OF THE TRIGGER ROLE OFPEROXYNITRITE IN THE ANTIARRHYTHMIC EFFECT OF ISCHAEMIC PRECONDITIONING ANDPEROXYNITRITE INFUSION
László Juhász1, Attila Kiss1, Edit Nyeső1, Mária Kovács1,
György Seprényi2, József Kaszaki3, Ágnes Végh1
Department of Pharmacology and Pharmacotherapy, University of Szeged
2
Department of Medical Biology, University of Szeged
3
Institute of Surgical Research, University of Szeged

1

Tutor: Prof. Dr. Ágnes Végh, University of Szeged
E-mail: jilajila@gmail.com

9Official Journal of the Romanian Society of Physiological Sciences
Background and purpose: The present study examines
whether peroxynitrite (PN) generated during the preconditioning(PC) procedure, or administered in brief intracoronary infusions
plays a trigger role in the antiarrhythmic effects of PC and PN in
aneasthetised dogs. Experimetal approach: The PN scavanger
urate (UA; 0.2 mg kg–1 min–1) was administered in PC (UA+PC;
n=8) and in PN treated (UA+PN; n=8) dogs or in control animals(UAC; n=9) over 30 min, just prior to a 25 min occlusion of theleft anterior descending (LAD) coronary artery. Severities ofischaemia (ST-segment elevation, inhomogeneity of electricalactivation) and ventricular arrhythmias (VPBs, VT, VF), plasma
nitrite/nitrate (NOx) levels as well as superoxide and myocardial
nitrotyrosine (NT) productions were determined. Key results:
Both the PC procedure (n=10) and the infusion of PN (n=10)
resulted in elevations in NT formation, and compared to thecontrols (C; n=14) markedly reduced the number of VPBs (276
± 85 vs 24 ± 10 and 55 ± 19), increased survival (S: 0% vs 66%
and 50%), preserved NOx formation and attenuated superoxide
and NT productions that resulted from a prolonged ischaemia
and reperfusion insult. Although UA inhibited NT formation inboth groups the protective effects were only abolished in PN(VPBs: 226 ± 121; S: 25%) but not in PC dogs (VPBs: 12 ± 6; S:62%). Interestingly, urate itself was also antiarrhythmic (VPBs: 50± 14; S: 55%). Conclusions and Implications: Low concentrationof PN can induce an antiarrhythmic effect but PN generated
during the PC procedure is not necessary for the PC-induced
antiarrhythmic protection.
Keywords: Preconditioning; peroxynitrite; urate; ischaemia/
reperfusion; arrhythmias

14. ACUTE ADMINISTRATION OF SIMVASTATIN
REDUCES THE SEVERITY OF VENTRICULAR ARRHYTHMIAS IN ANESTHETIZED DOGS
Gábor Kisvári1, Mária Kovács1, József Kaszaki2 and Ágnes
Végh1
Department of Pharmacology and Pharmacotherapy, University of Szeged
2
Institute of Surgical Research, University of Szeged

1

Tutor: Prof. Dr. Ágnes Végh, University of Szeged
E-mail: kisvari.gabor@gmail.com
During the past decade statins were found to have a pleiotropic
effect beyond their lipid-lowering ability. These effects, especiallythat occur following acute drug administration and may contributeto the cardioprotection, are still poorly understood. Our aim was
to investigate the effect of a single bolus injection of simvastatin(0.1 mg/kg) on the ischaemia and reperfusion-induced ventriculararrhythmias in chloralose-urethane-anaesthetized dogs. In one
group of dogs (n=7) activated simvastatin, in the other group(n=8) the solvent of the compound were given just prior to a 25
min occlusion of the left anterior descending coronary artery.
The severity of ischaemia (degree of inhomogenity of electricalactivation, epicardial ST-segment) and of arrhythmias, as well as
the plasma nitrate/nitrite (NOx) levels were assessed during theocclusion. Compared to the control group, simvastatin decreased
the total numbers of VPBs (206 ± 44 vs. 139 ± 50) and episodes of10

VT (5.8 ± 1.9 vs. 0.5 ± 0.3), the incidences of VT (70% vs. 38%) andVF (40% vs. 0%) during the occlusion. Survival from the combined
ischaemia/reperfusion was also higher (38%) in the simvastatin
treated group than in the controls (0%). Whereas in the control
group the plasma level of NO metabolites in the coronary sinus
blood was markedly reduced, in the simvastatin treated animals
NOx levels were significantly increased particularly during thesecond phase of the occlusion. We conclude that acute simvastatin
treatment may provide protection against the ischaemia andreperfusion-induced severe ventricular arrhythmias perhaps
through the elevation of NO availability during ischaemia. This
work is supported by the Hungarian Scientific Research Foundation(OTKA, project No. K75281).
Keywords: simvastatin, ischaemia, ventricular arrhythmias,NO

15. EFFICIENCY OF PLASMID TRANSFECTION ALONG
THE REGENERATING RAT SOLEUS MUSCLE
Magdolna Kósa, Ernő Zádor
University of Szeged, Faculty of General Medicine, Departmentof Biochemistry
Tutor: Prof. Dr. Ernő Zádor, University of Szeged
E-mail: magdi@biochem.szote.u-szeged.hu
Regeneration plays an important role in skeletal muscle
diseases and injuries. Experimental regeneration can be induced
in different ways; we use direct intramuscular injection of snake
venom (notexin) to achieve a complete necrosis and subsequent
regeneration in the rat soleus muscle. The regenerating soleus
can be successfully transfected by naked plasmid injection.
However this procedure results in a low efficiency transfection
which has not been characterized along the entire length ofthe muscle.
The regenerating soleus muscles were transfected with plasmids
encoding green or red fluorescent proteins. After one week themuscles were dissected, frozen and cut into 6 approximately
equal segments before frozen sectioning. Transversal sections
were studied with fluorescent microscope for transfected fibres
in each segment. The neuromuscular endplates were also stainedin each muscle segment for determination of fibre length. We
found transfections mostly in the central segments, near to thesite of intramuscular injection. The transfected fibres were usuallyorganized in groups, next to each other on cross sections. The
neuromuscular endplates were also found mostly in the central
segments. Considering that each fibre has one endplate, thisconcentration suggests that most fibres run from proximal
to distal ends along the entire muscle. From these results weconclude that the transfection efficiency is even further reduced
along the fibres in the regenerating muscle. This is not due toreduced fibre length, since most fibres extend from proximal
to distal ends of the muscle, but rather caused by the limited
migration of the transfected plasmids and expressed proteins
along the fibres.
Keywords: muscle regeneration, transfection distribution,
fibre length

Fiziologia ‑ Physiology  2011 supplement

Official Journal of the Romanian Society of Physiological Sciences

16. SPECIFIC CHEMOKINE-CHEMOKINE RECEPTORS INTERACTIONS AND THEIR ROLE IN hMSC
MIGRATION

17. VENTRICULAR ASYNCHRONISM - PROGNOSIS MARKER IN HEART FAILURE WITH PRESERVEEJECTION FRACTION

Marusciac Laura

Adelina Mavrea, Ioana Citu, Mirela Tomescu

Victor Babes University of Medicine and Pharmacy Timisoara,
Department of Physiology-Immunology

Victor Babes University of Medicine and Pharmacy Timisoara,
Department of Medical Semiology

Tutors: Prof. Dr. Virgil Paunescu, Victor Babes University of
Medicine and Pharmacy Timisoara; Prof. Reinhard Henschler,
Institute of Transfusion Medicine and Immune Hematology, Blood
Donation Service of the German Red Cross, Frankfurt, Germany
E-mail: laura.marusciac@gmail.com
Background and aims: Human mesenchymal stem
cells (hMSCs) are non-hematopoietic stromal cells that candifferentiate into different tissues. They are rare in the bonemarrow (0.001-0.01% of nucleated cells). Minimal criteria havebeen established to define hMSCs. They have leukocyte-likefunctions, expressing various chemokine receptors, integrins,
and adhesion molecules, and displaying migratory capacity
towards ligands released from injury sites. The aims are toisolate, characterize, differentiate hMSCs, to determine theirmigratory signals, and the in vitro & in vivo migration of hMSCs
to specific chemokines.
Material and methods: hMSCs were isolated from humanbone marrow using the density gradient method. After 2 weeks
expansion, they were characterized using flow cytometry. The
hMSCs were then cultured in adipogenic, chondrogenic, andosteogenic self-made media for 3 weeks, and stained with specificstains for adipose tissue (Oil Red O), bone (Alizarin Red), andcartilage (Alcian Blue). The surface and intracellular expressionof 7 pro-inflammatory chemokines and 13 chemokine receptors
were analyzed on undifferentiated hMSCs.
Results: hMSCs have been successfully isolated from 8bone marrow samples, obtaining an average of 110 x 10 6 bonemarrow cells after isolation. They have been expanded andkept in culture for up to 6 passages. They fulfilled the minimal
criteria for hMSCs (expression of CD105, CD73, and CD90,
lack of CD11a, CD34, and CD45 expression). The hMSCs weresuccessfully differentiated into three lineages using self-mademedia. There was no chemokine receptor surface expression,
due to trypsinization. The chemokine receptor intracellular
expression was donor-dependent, with the constant expressionof CCR3, CCR4, and CXCR3. The chemokine intracellular
expression was also heterogenic, only CCL2 and CXCL12 being
constantly expressed.
Conclusion: The expression of chemokines and chemokine
receptors in healthy donors is highly heterogenic. There are
specific interactions between chemokines and chemokine
receptors during inflammation, and these specific interactions
influence hMSC migration.
Keywords: hMSC, chemokines, chemokine receptors,
migration, inflammation

Tutor: Prof. Dr. Mirela Tomescu, Victor Babes University of
Medicine and Pharmacy Timisoara
E-mail: amavrea@yahoo.com
It is now recognized that at least 40% of patients with heart
failure have normal ejection fraction (FE). If left ventricular(LV) systolic function is normal, it may be difficult to make aconclusive determination of the relative role of diastolic heart
failure compared with other concomitant conditions.
I propose to investigate the association between ventricularasynchrony, LV diastolic dysfunction and other biomarkers ofprognosis in diastolic heart failure
The objectives are to identify new prognostic markers that
correlate with clinical and cardiac functional exploration, toassess the progression of heart failure, prognostic stratificationof heart failure and identifying patients at high risk ofcardiovascular events.
Inclusion criteria: clinical symptoms of heart failure NYHA
II, III; sinus rhythm; FE > 45%.
Exclusion criteria: atrial fibrillation, relevant valve disorders,
hypertrophic cardiomyopathy and severe lung disease.
For echocardiography diagnosis of diastolic heart failure
the following are determined: mitral Doppler profile, theprofile of pulmonary venous flow Doppler, early diastolic
flow propagation velocity and myocardial performanceindex (GMI).
Ventricular asynchronism is assessed by the ratio of left
ventricular filling time and cardiac cycle, determining theinterventricular mechanical delay (IVMD), Septal – posteriorwall motion delay (SPWMD), aortic preejectional time. Usingtissue Doppler the differences between peak systolic velocitiesof opposite walls will be measured.
All patients will be physically and echocardiography examined,
6-minute-walk-testing will be assessed and determinationof serum prognostic markers of heart failure (serum pro-BNP,
serum troponin, serum D-dimmer) will be performed.
After one year we will reevaluate the parameters used forthe study and we will compare patients’ evolution with andwithout ventricular asynchrony and the presence of prognosis
biomarkers.
With multimarker strategy
I intend to identify the combinations of biomarkers andechocardiography parameters that are optimal at various
stages during the evolution of diastolic heart failure, ranging
from their use for screening, diagnosis, determining prognosis,
and guiding management.
Keywords: diastolic dysfunction, heart failure, asynchronism,
marker prognosis

Fiziologia ‑ Physiology  2011 supplement

11Official Journal of the Romanian Society of Physiological Sciences

18. MESOSTRUCTURE. TYPES OF BARS IN IMPLANT OVERDENTURE
Naiche Diana*, Baldea Bogdan*, Bratu Dorin**, Candea
Adrian**Prosthodontic Department, Faculty of Dentistry, UMF Timisoara**Prosthodontic Department, Faculty of Dentistry, UMF Timisoara
Tutor: Prof. Dr. Dorin Bratu, Victor Babes University of Medicine
and Pharmacy Timisoara
E-mail: diana_naiche80@yahoo.com
Prosthetic rehabilitation of totally edentulous patients,
especially on the mandibular jaw, with complete dentures is nolonger the first treatment method choice. After the Consensus
held in 2002, at McGill University, Canada, it was decided thatthe standard of care for this category of patients is represented
by the overdenture supported on 2 interforaminal implants. In
time, this treatment concept became viable and other treatment
solutions have emerged. The best known classification of prosthetic
implant options is that one proposed by C.E. Misch. The numberof implants inserted varies for each patient, depending on its
financial availability and anatomical limitations. The connection
between the implant and overdenture is done through different
connection systems: balls, bar / rider, magnets and telescopic
systems. In case of using implant bar overdentures, we candistinguish three levels at this type of prosthetic restorations:infrastructure, mesostructure, and superstructure. There areseveral types of bars used for implant overdentures, and the
purpose of this presentation is to expose them: cemented
and screwed bars, short and long bars, intraorally weldedprefabricated bars, metallic (titanium) milled bars, zirconium
milled bars.
Keywords: mesostructure, zirconium, overdenture

19. EPIDEMIOLOGY OF CENTRAL NERVOUS SYSTEM
MALFORMATIONS IN A REGION IN HUNGARY
Nóra Szabó1, Gyurgyinka Gergev1, Jenő Kóbor1, András
Svékus2, Edit Bereg1, Péter Szűcs1, Éva Romhányi1, Andrea
Valek3, László Kaizer4, Sándor Túri1, László Sztriha1
University of Szeged, Department of Pediatrics
Municipial Hospital, Gyula
3
National Centre for Healthcare Audit and Improvement, National
Congenital Abnormality Registry, Budapest
4
University of Szeged, Department of Pathology

12

Tutor: Prof. Dr. László Sztriha, University of Szeged
E-mail: szabo.nora.zs@gmail.com
Background: Few data are available on the epidemiologyof central nervous system (CNS) malformations despite theirmajor aetiological role in mental and neurological handicap
and epilepsy in childhood.
Aim of the study: To describe the population-based
epidemiological characteristics and spectrum of various
malformations of the CNS in South-Eastern Hungary and compile12

a registry.
Methods: A retrospective survey of patients born with CNS
malformations in the region between July 1, 1992 and June 30,
2006 was performed. The malformations were classified into 22
subgroups and the livebirth prevalence per 10 000 births wascalculated.  
Results and Conclusions: CNS malformations were foundin 214 children among 185 486 livebirths, which correspond toa livebirth prevalence of 11.54 per 10 000. Neural tube defects(spina bifida, anencephaly, encephalocele) and corpus callosum
anomalies were the most common malformations with livebirthprevalence figures of 2.21 and 2.05 per 10 000 births.
The livebirth prevalence of holoprosencephaly, septo-optic
dysplasia, cavum septum pellucidum, microcephaly, microlissencephaly,
lissencephaly-agyria/pachygyria and subcortical band heterotopia
spectrum, neuronal heterotopia, polymicrogyria, schizencephaly,
focal cortical dysplasia, Joubert syndrome, unilateral cerebellar
hypoplasia, Dandy-Walker malformation, cerebellar vermis
hypoplasia, cerebellar aplasia/hypoplasia, Chiari I malformation,
pontocerebellar hypoplasia and arachnoid cysts was 0.49, 0.11,
0.59, 1.62, 0.11, 0.27, 0.16, 0.43, 0.54, 0.05, 0.05, 0.11, 0.32, 0.49,
0.38, 0.11, 0.16, and 1.29 per 10 000 livebirths, respectively.
Somatic anomalies in association with the CNS dysgenesis
were observed in 39.2% of the patients. Seizures appeared
and epilepsy was diagnosed in 35.9%. Developmental delay,
intellectual disability or abnormal neurological signs occurredin more than half of the patients.
Our registry of CNS malformations provides a database forfurther studies on the genetic and environmental aetiology of
CNS dysgenesis.
Keywords: epidemiology, central nervous system malformations,
livebirth prevalence

20. STUDY ON THE LEVELS OF TAU AND β-AMYLOID INCSF OF PATIENTS WITH MULTIPLE SCLEROSIS
Levente Szalardy, Denes Zadori, Agnes Koszo, Krisztina
Bencsik, Laszlo Vecsei, Peter Klivenyi
Department of Neurology, University of Szeged
Tutor: Dr. Peter Klivenyi, University of Szeged
E-mail: levente.szalardy@yahoo.com
Multiple sclerosis (MS) is a progressive central nervous system(CNS) disorder characterized inflammation, demyelination, axonal
damage and neurodegeneration. The course of the disease canvary in a wide range and finding a proper way to predict diseaseprogression is a matter of great significance.
Our objective was to examine if the cerebrospinal fluid (CSF)
levels of proteins previously proved to be relevant in common
neurological diseases could predict the progression and future
clinical manifestation of MS.
We evaluated the levels of h-Tau, p-Tau and β-amyloid inhuman CSF samples collected from patients presenting withdifferent clinical forms of MS by commercially available ELISA kits.
The lumbar punctures were performed from 1999 to 2007 andpatients underwent regular clinical follow-up and were classified
as clinically isolated syndrome (CIS), relapse remitting form (RR)

Fiziologia ‑ Physiology  2011 supplement

Official Journal of the Romanian Society of Physiological Sciences
and primary progressive MS (PP).
We did not find any difference in the examined protein levels
between MS and control groups. No significant difference could beobserved between groups with different progression tendencies;
however, there was an observable decrease in p-Tau concentrationin the primary progressive group, which did not reach the levelof significance.
The examined proteins seem to be inappropriate as biomarkers
in predicting disease progression in MS. More focus should be
devoted to the examination of primary progressive MS with anincreased number of patients involved.
Key words: sclerosis multiplex, demyelination,
biomarker, ELISA

21. INVESTIGATION OF TRANSPORTER INTERACTIONS OF ANTIMALARIALS IN VITRO
Péter Szerémy1,2, Ildikó Makai1, Márton Jani1, Marton
Lívia1, Szilvia Gedey1, Katalin Jakab1, János Márki-Zay1,2,
Péter Krajcsi1
Solvo Biotechnology, Szeged
Department of Biochemistry, Faculty of Medicine, Universityof Szeged12

Tutor: Peter Krajcsi, Solvo Biotechnology, Szeged
E-mail: szeremy@solvo.com
Options to control spread of malaria are increasingly limited
due to emergence of parasites resistant to widely used antimalarials,
therefore, discovery of novel antimalarials appears crucial as ever.
However, animal experiments are too expensive and laborious
for the pharmacokinetic characterization of large number ofcompounds. The fate of administered drugs may largely dependon their interactions with transporter proteins, which are present
in all major pharmacologically relevant barriers. Furthermore,transporters are key determinants of antimalarial drug resistanceof plasmodiums as well. The aim of this study was to examine
whether the high-throughput (HTS) cell and membrane-basedtransporter assays can be applied to characterize the transporter
interactions of candidate antimalarials.
Reference antimalarials, such as artemisinin, chloroquine,
mefloquine, quinine, etc, have been tested for their interaction
with the ABC-transporters MDR1, MRP1 and BCRP using the Solvo
PredEasy ATPase kits and the interaction with the SLC family
members OCT1 and OCT2 uptake transporters in cell-based
assay. Measured IC50 and EC50 values were correlated with theclinical observations on the tested antimalarials.
In case of Amodiaquine, Hydroxychloroquine, Primaquine,
Pyrimethamine, Proguanil, Artemisinin, Artesunate, Atovaquone,
Clindamycin, Halofantrine, our data are the first proof for transporter
interaction of these clinically important drugs. Artemisinin is a
substrate for MDR1, chloroquine is inhibitor of the MDR1 andsubstrate for the MRP1 and BCRP, mefloquine is substrate for
the MDR1 but at higher concentrations is a not specific inhibitorof all the transporters and quinine is substrate for the MDR1.
These results corresponded exactly to the clinical data on theantimalarials tested.
We conclude that the membrane- and cell-based HTS in vitro

Fiziologia ‑ Physiology  2011 supplement

assays can be applied to facilitate the ADME characterization ofcandidate antimalarials.
Keywords: antimalarial, transporter interaction, ADME, invitro assays

22. ISCHEMIC LIMB SALVAGE USING NEOANGIOGENIC THERAPY – CASE REPORT
Georgel Taranu1, Andreea Rata1, Liviu Tamas2, Anca Tursie1,
Sylvana Gyenes1, Andrei Anghel2
Vascular Surgery Department, Emergency County Hospital
Timisoara2
Biochemistry Department, University of Medicine and Pharmacy“Victor Babes” Timisoara
1

Tutor: Prof. Dr. Andrei Anghel, Victor Babes University of Medicine
and Pharmacy Timisoara
E-mail: gptgeorgel@yahoo.com
The present paper describes the case of a 50-year old patient,heavy smoker, diagnosed at the age of 48 with peripheral occlusive
arterial disease (occlusion of the left superficial femoral artery),when a revascularization procedure was attempted (not possible
because of extensive thrombosis in the popliteal artery andpoor distal arterial bed – highly suggestive for thromboangiitis
obliterans). In the next period the patient developed a tissue
loss in the calf; skin graft was performed (in some other surgicaldepartment) but with no result. Considering the current symptoms(rest pain), the peripheral angiographic aspect and the trophic
lesions in the calf, in September 2008 the first gene therapy
procedure was performed, followed by repeated administration
at one month. The patient was monitored for the next 18 monthand the clinical course was good, with relief of rest pain andsecondary healing of trophic lesions.
Keywords: neoangiogenic therapy, VEGF, HGF, critical limb
ischemia

23. PROTECTIVE EFFECT OF L-α GLYCERYLPHOSPHORYLCHOLINE AGAINST ISCHAEMIA-REPERFUSION INJURY IN THE RAT SMALL INTESTINE
Tünde Tőkés, Gabriella Varga, Eszter Tuboly, László Major,
József Kaszaki, Miklós Ghyczy, Mihály Boros
University of Szeged, Institute of Surgical Research, Szeged,
Hungary
Tutors: Dr. József Kaszaki; Prof. Dr. Mihály Boros, Universityof Szeged
E-mail: tunde@expsur.szote.u-szeged.hu
Introduction: Our previous studies showed that phosphatidylcholine(PC) inhibits the production of reactive oxygen species (ROS) bothin vitro and in vivo, and displays anti-inflammatory features during
ischaemia-reperfusion (IR) conditions. Our present aims were to
examine whether the anti-inflammatory effects of PC are linked
to the fatty acids of the molecule or the headgroup, therefore
we studied the effects of L-α glycerylphosphorylcholine (GPC),
the deacilated polar derivative of PC, in a rat model of IR.
Methods: Anesthetized rats were randomized to control

13

Official Journal of the Romanian Society of Physiological Sciences(n=8), mesenteric IR (n=8), IR with GPC pre-treatment (n=8)
or IR with GPC post-treatment (n=8) groups. We monitored
the macrohaemodynamics (mean arterial pressure, arteria
mesenterica superior flow, mesenteric vascular resistance(MVR) and microhaemodynamic parameters (serosal blood
flow, with intravital OPS videomicroscopy) and biochemicalmarkers (tissue superoxide (O2.-), xanthine oxidoreductase (XOR)
activity, and liver ATP-contents, after 45 min ischemia and 180
min reperfusion period.
Results: The IR insult increased MVR and tissue O2.- production,and reduced liver ATP content after reoxygenation. Nevertheless,
GPC pre-treatment reduced the elevation of MVR and decreased
the tissue O2.- production, while post-treatment diminished both
XOR activity and ATP depletion, and normalized the intestinal
microcirculatory dysfunction.
Conclusion: Exogenous GPC has anti-inflammatory effectand decreases IR-induced ROS production. The similar efficacyof PC could be linked to a reaction involving the polar part ofthe molecule.
Supported by: OTKA K75161; TAMOP-4.2.1; TÁMOP-4.2.2;
ETT 442/2009
Keywords: Mesenteric vascular resistance (MVR), superoxide,
xanthine oxidoreductase, ATP

24. CHARACTERIZATION OF THE EFFECTS OF
TWO PORPHYRIN COMPOUNDS ON RAT HEART
MITOCHONDRIAL RESPIRATION
Simona Trancotă1, Corina Enache2, Oana Duicu1, EugeniaFăgădar-Cosma2, Danina Muntean1
Department of Pathophysiology, “Victor Babeş” Universityof Medicine and Pharmacy 2Institute of Chemistry-RomanianAcademy, Timişoara, Romania

1

Tutor: Prof. Dr. Danina Muntean, Victor Babes University of
Medicine and Pharmacy Timisoara
E-mail: mitica_simona_lavinia@yahoo.com
Mitochondria play a central role in health and disease, andcompounds such as metallo-porphyrins, able to enter mitochondria,
are nowadays intensively investigated. The aim of the present
study was to assess the effects of two porphyrins: Zn(II)-5,10,15,20tetrakis(4-pyridyl)porphyrin (ZnTPyP) and 5,10,15,20-tetrakis(4-allyloxyphenyl) porphyrin (TAPP) on respiratory function of rat
heart mitochondria isolated by means of differential centrifugation
technique. Oxygen consumption of freshly isolated mitochondria(0.5 mg/mL, final concentration of mitochondrial proteins) was
measured using a Clark-type respirometer at 37ºC in a sucrose
buffer (pH 7.2) in the presence of NAD-linked (glutamate/malate)and FAD-linked (succinate) substrates, respectively. Basal (state 2)
and ADP-stimulated (state 3) respiratory rates were recorded andexpressed as nanoatoms oxygen/min/mg protein and respiratory
control index (RCI) was calculated. Three concentrations (3, 5
and 10 μM, respectively) of each porphyrin were studied. In thepresence of complex I substrates (glutamate/malate), ZnTPyP(5 µM final concentration) elicited a significant increase ofstate 2 and state 3 respiratory rates (70.5±2.2 and 468±22.6, p <0.0001 vs. Control) but had no effect on complex II dependent14

respiratory rates. However, in the presence of TAPP (10 µM final
concentration) oxidation of glutamate and malate was diminished:state 3 respiratory rate was decreased by 29% and RCI by 31%.
TAPP (10 µM) also decreased basal respiration by 16% in the caseof succinate-supported respiration. In normal isolated rat heart
mitochondria Zn(II)-5,10,15,20-tetrakis(4-pyridyl) porphyrin and
5,10,15,20-tetrakis(4-allyloxyphenyl) porphyrin elicited different
effects on complex I and complex II supported respiration, an
observation that clearly warrants further investigation view theirpotential application in pathological conditions associated withischaemia/reperfusion injury.
Keywords: mitochondria, metallo-porphyrins, rat,
respiration

25. The effects of exogenous methane
inhalation on macro-and microcirculatory changes during intestinal ischemiareperfusion in rats
Eszter Tuboly, Gabriella Varga, Tünde Tőkés, József Kaszaki,
Miklós Ghyczy, Mihály Boros
University of Szeged, Institute of Surgical Research, Hungary
Tutors: Prof. Dr. Mihály Boros; Dr. József Kaszaki, Universityof Szeged
E-mail: eszter.tuboly4@gmail.com
Introduction: The small intestinal mucosa is particularly
sensitive to ischemia-reperfusion (I/R) injuries. I/R initiates acomplex biochemical cascade reaction in which the generation ofreactive oxygen species (ROS) and the micro-and macrocirculatory
perfusion deficiencies will lead to tissue damage and barrier
function failure. Mammalian methanogenesis is considered an
exclusive indicator of carbohydrate fermentation of the anaerobic
gastrointestinal flora, but in our previous studies we haveshown preventive effects of methane on hypoxia-induced
inflammatory processes (Ghyczy et al., FASEB J 2003; Cell Physiol
Biochem 2008). Based on this background, we hypothesizedthat the damaged intestinal microperfusion during I/R could
be improved by exogenous methane inhalation.
Methods: Male Sprague-Dawley rats were randomized tocontrol (n=7), I/R without methane (I/R, n=7) and I/R with 2.5%
methane added to 21% oxygen (n=7) groups. The superior
mesenteric artery (SMA) was occluded for 45 min, which was
followed by 120 min reperfusion. Methane treatment was started
during the last 10 min of ischemia and lasted for 5 min during
reperfusion. Systemic and mesenteric haemodynamics weremonitored, the microcirculation of the serosa of the terminal
ileum was observed by intravital videomicroscopy (IVM) using
orthogonal spectral polarisation imaging technique (A/R Cytoscan).
The tissue ROS generation was detected by a chemiluminescenceassay, the myeloperoxidase (MPO) activity was assessed byfluorimetric analysis.
Results: The significant microcirculatory dysfunction (asevidenced by IVM) after I/R was remarkably improved by methane
treatment. Moreover, a significantly decreased superoxide
production and MPO activity were detected in the treated animals,
in contrast with the control group.

Fiziologia ‑ Physiology  2011 supplement

Official Journal of the Romanian Society of Physiological Sciences
Discussion: Exogenous methane improves the perfusion
deficiency of the mucosa after I/R, and this effect may be linked
to ROS scavenging or inhibition of leukocyte activation in thereperfused intestinal tissues.
Keywords: intestinal ischemia, reactive oxygen species,
microcirculation, leukocyte activation
Supported by TÁMOP-4.2.2-08/1-2008-0013

26. CHILDBEARING AMONG WOMEN WITH MULTIPLE SCLEROSIS (MS)Melinda Vanya
University of Szeged, Department of Obstetrics and Gynecology
Tutor: Prof. Dr. György Bártfai, University of Szeged
E-mail: vmelinda74@gmail.com
Introduction: Pregnancy management issues an extra
challenge to MS patients and their physicians. There arefew epidemiological studies reporting databases on thissubject.
Objectives: To report the results between 1998
and 2010 from the University of Szeged, Department of
Obstetrics and Gynecology, Department of Neurology
database on multiple sclerosis and pregnancy. Patients

Fiziologia ‑ Physiology  2011 supplement

and methods: A retrospective analysis of pregnancy
data from MS patients, who became pregnant at any
time of their disease, compared to international research
was carried out. Results: In 65 MS mothers, the average
age at pregnancy was 35.87 (± 7.9). EDSS at start pregnancy
was 1.38(±1,4) and relapse rate before antepartum period was
1.37(±1.3). The average number of pregnancies was 2.14(SD±1.3), the average fetal weight: 3106.1±630 g. Exposure
to MS drugs at any time during pregnancy was high (67%)
In our study the overall spontaneous abortion rate wassignificantly higher in MS patients, as the age-matchedhealthy control (p = 0.0006). Vaginal delivery incidence
was relatively high (67.9%) Incidence of Cesarean section
was 35.4%, and 33.8% was the incidence of interruption.
The prevalence of pregnancy complications was relatively
low with 12.3 % premature births outcomes. Metrorrhagia
was seen in 26. 1% and myomas were seen in 13.8%.
Conclusion: Our results confirm previous findings
lower relapse rate during pregnancy and add to present
literature informing on data related to immunmodulatory
treatment, spontaneous abortion, changes of BMI andobstetrical complications.
Keywords: immunmodulatory treatment during pregnancy,
spontaneous abortion, obstetrical complications

15

Official Journal of theautHorS
Romanian Societyof Physiological Sciences
index

BALINISTEANU, B.............................................5KISVARI, G.......................................................10

BERE, ZS..............................................................5

KOSA, M............................................................11

BOCAN, V............................................................5

MARUSCIAC, L................................................11

BORDA, B............................................................6

MAVREA, A......................................................11

BOTA, S...............................................................6

NAICHE, D.........................................................12COMSA, S............................................................6SZABO, N..........................................................12

CORICI, C............................................................7

SZALARDY, L...................................................12

DUSE, A...............................................................7

SZEREMY, P......................................................12

FEFLEA, S............................................................8

TARANU, G.......................................................13GARAB, D............................................................8

TOKES, T...........................................................13

GOMBAR, CS......................................................9

TRANCOTA, S..................................................13

GRUICI, A............................................................9

TUBOLY, E........................................................14

JUHASZ, L.........................................................10

VANYA, M........................................................14

16

Fiziologia ‑ Physiology  2011 supplement