programme and abstract book - Ústav klinické a molekulární ...

PROGRAMME AND ABSTRACT BOOK 

APRIL 29–30, 2011 

OLOMOUC 

THE CZECH REPUBLIC 

ISBN 978-80-87327-59-3 

The 7th Symposium & Workshop 

on Molecular Pathology 

and Histo(cyto)chemistry 

The 96th Seminar 

of the Czech Division 

of the International Academy 

of Pathology 

The 3rd Olomouc Days 

of Histology Laboratory Technicians 

ORGANIZED BY 

The Czech Society of Pathologists CLS JEP 

The Molecular Pathology Working Group of the Czech Society 

of Pathologists and the European Society of Pathology 

The Czech Oncological Society CLS JEP 

The Czech Society for Histochemistry and Cytochemistry 

The Czech Society of Laboratory Technicians 

The Department of Pathology & the Laboratory of Molecular 

Pathology; The Laboratory of Experimental Medicine, Faculty of 

Medicine and Dentistry, Palacký University, Olomouc 

University Hospital, Olomouc

The 7th Symposium & Workshop on Molecular Pathology and Histo(cyto)chemistry 

The 96th Seminar of the Czech Division of the International Academy of Pathology 

The 3rd Olomouc Days of Histology Laboratory Technicians 

April 29–30, 2011, Olomouc, Czech Republic 

Under the auspices of 

prof. M. Mašláň, Ph.D. / Rector of Palacký University, Olomouc 

prof. Z. Kolář, M.D., Ph.D. / Dean of the Faculty of Medicine and Dentistry, Palacký University, Olomouc 

R. Maráček, M.D. / Director of the University Hospital Olomouc 

Chairman of the Congress: 

Professor Zdeněk Kolář, M.D., Ph.D. / The Department of Pathology & the Laboratory of Molecular Pathology 

Faculty of Medicine and Dentistry, Palacký University, Olomouc, Hněvotínská 3, Olomouc, 775 15 

Tel: +420/585 632 451; Fax: +420/585 632 966; e-mail: kolarz@tunw.upol.cz 

Organizing & Programme Committee: 

Professor Jiří Ehrmann, M.D., Ph.D. / The Department of Pathology & the Laboratory of Molecular Pathology 

Faculty of Medicine and Dentistry, Palacký University, Olomouc, Hněvotínská 3, Olomouc, 775 15 

Tel: +420/585 632 466; Fax: +420/585 632 966; e-mail: info.lmp@seznam.cz 

Associate professor Martin Tichý, M.D., Ph.D. 

The Department of Pathology, Faculty of Medicine and Dentistry Palacký University, Olomouc, 

University Hospital, Hněvotínská 3, Olomouc, 775 15 

Tel: +420/585 632 453; Fax: +420/585 632 966; e-mail: info.lmp@seznam.cz 

Danuše Kvapilová 

The Department of Pathology, University Hospital, Hněvotínská 3, Olomouc, 775 15 

Tel: +420/585 632 465; Fax: +420/585 632 966; e-mail: d.kvapilova@post.cz 

Jan Bouchal, Marie Geierová, Marián Hajdúch, Jana Holinková, Monika Levková, Bohuslav Melichar, Eva Pimrová, 

Lenka Prokopová, Jana Steigerová, Jozef Škarda, Michaela Šváchová, Ivo Überall 

Venue: 

Friday, April 29 – Regional Centre Olomouc (Jeremenkova 40B, Olomouc, 772 00) 

Saturday, April 30 – Theoretical Institutes Building, Faculty of Medicine and Dentistry, Palacký University, Olomouc 

(Hněvotínská 3, Olomouc, 775 15) 

Conference Language: 

The 7 th Symposium & Workshop on Molecular Pathology and Histo(cyto)chemistry – English 

The 96 th Slide Seminar of the Czech Division of the International Academy of Pathology – Czech 

The 3 rd Congress of Histology Laboratory Technicians – Czech 

APRIL 29–30, 2011 | OLOMOUC | THE CZECH REPUBLIC 

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PROGRAMME 

The 7 th Symposium on Molecular Pathology and Histo(cyto)chemistry 

FRIDAY, APRIL 29, 2011 

Time Venue / language 

8:30–9:00 Registration 

9:00–9:15 Joint meeting ceremony Regional Centre 

Centaurus Hall / English, Czech 

Keynote lectures of invited speakers I 

Chairs: P. Dubový (Brno), E. Pikarski (Jerusalem) 

9:15–9:45 P. Dubový (Brno) 

Expression of cytokine proteins and mRNAs in the primary sensory neurons 

of neuropathic pain model 

9:45–10:15 E. Pikarski (Jerusalem) 

Advances in molecular diagnostics of hepatocellular carcinoma 

10:15–10:45 J. Moreira (Copenhagen) 

Development of novel early detection and stratifi cation markers for breast 

cancer: integrating pathology and proteomics 

10:45–11:15 L. Havel (Brno) 

Histochemistry in plant development and abiotic and biotic reactions 

11:15–11:45 Coff ee break 

Tissue Banking 

Chairs: P. De Blasio (Milano), L. Dušek (Brno) 

11:45–12:05 P. De Blasio (Milano) 

Tissue Banking and Tissue Microarrays 

12:05–12:25 L. Dušek (Brno) 

How to manage real time data support for registries and tissue banking in 

clinical practice? Cancer care case study 

12:25–12:45 G. Stanta (Trieste) 

Archive tissue biobanking as a basis for translational and reverse translational 

research 


Regional Centre 

Perseus Hall / English 








Centaurus Hall / English 





12:45–13:00 Discussion Regional Centre 


13:00–14:00 Lunch break 

Poster Session 

Keynote lectures of invited speakers II 

Chairs: P. Murray (Birmingham), K. Smetana (Prague) 

14:00–14:30 P. Murray (Birmingham) 

Small lipids as therapeutic targets in cancer 

14:30–15:00 K. Smetana (Prague) 

Social life of cancer cell 

15:00–15:30 J. Sikora (Prague) 

Danon disease – lysosomal dyfunction disease – molecular pathology 

of LAMP2 protein - implications for clinical diagnostics 

15:30–15:45 R. C. Ecker (Wien) 

Applications of Slide-Based Single Cell Cytometry by TissueFAXS 

in Histopathology 









15:45–16:00 Discussion Regional Centre 

Perseus Hall / English

POSTERS 

PROGRAMME 

1 Increased susceptibility of MMP19 defi cient mice to DSS-induced colitis provides novel insights into pathogenesis 

of intestinal infl ammation 

Brauer R. 1 , Dziechciarkova M. 2 , Skarda J. 3 , Hajduch M. 2 , Sedlacek R. 1 

1Department of Transgenic Models of Diseases, Institute of Molecular Genetics AS CR, v.v.i, Prague, Czech Republic 

2Laboratory of Experimental Medicine, Department of Pediatrics and Oncology, Medical Faculty, Palacky University, Olomouc, Czech Republic 

3 Department of Pathology, Medical Faculty, Palacky University, Olomouc, Czech Republic 

2 Quadriplex model enhances urine-based detection of prostate cancer 

Jamaspishvili T. 1 , Kral M. 2 , Khomeriki I. 2 , Vyhnankova V. 2 , Mgebrishvili G. 1 , Student V. 2 , Kolar Z1 . and Bouchal J. 1 

1 Laboratory of Molecular Pathology and Department of Clinical and Molecular Pathology, Institute of Molecular and Translational Medicine, 

Faculty of Medicine and Dentistry, Palacky University and University Hospital, Olomouc, Czech Republic 

2Department of Urology, University Hospital, Olomouc, Czech Republic 

3 Comparison of proliferating activity in terminal villi of normal and diabetic placenta 

Jirkovská M. 1 , Kučera T. 1 , Jadrníček M. 1 , Niedobová V. 1 , Moravcová M. 2 , Krejčí V. 2 , Žižka Z. 2 

1Institute of Histology and Embryology, First Faculty of Medicine, Charles University in Prague, Prague, Czech Republic 

2Department of Obstetrics and Gynecology, First Faculty of Medicine, Charles University in Prague, Prague, Czech Republic 

4 A novel transgenic mouse model to study epidermal diff erentiation and wounding 

Kasparek P., Suchanova S., Krenek P., Sedlacek R. 

Department of Transgenic Models of Diseases, Institute of Molecular Genetics AS CR, Prague, Czech Republic 

5 Asporin modulates tissue microenvironment, invasive growth and bone metastasis of breast cancer 

Kharaishvili G. 1 , Cizkova M. 2 , Bouchalova K. 2 , Sedlakova E. 1 , Kolar Z. 1 , Bouchal J. 1 

1 Laboratory of Molecular Pathology of Institute of Pathology, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, 

Palacky University, Olomouc, Czech Republic 

2Laboratory of Experimental Medicine, Pediatric Clinics of Medical Faculty of Palacky University and Faculty Hospital, Olomouc, Czech Republic 

6 Collagen triple helix repeat containing 1 protein in primary and metastatic breast cancer 

Kharaishvili G. 1 , Cizkova M. 2 , Bouchalova K. 2 , Sedlakova E. 1 , Kolar Z. 1 , Bouchal J. 1 

1 Laboratory of Molecular Pathology of Institute of Pathology, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, 

Palacky University, Olomouc, Czech Republic 

2Laboratory of Experimental Medicine, Pediatric Clinics of Medical Faculty of Palacky University and Faculty Hospital, Olomouc, Czech Republic 

7 Apoptosis in placental vessels from pregnancies complicated by diabetes mellitus type I 

Kučera T. 1 , Jadrníček M. 1 , Niedobová V. 1 , Žižka Z. 2 , Krejčí V. 2 , Moravcová M. 2 , Jirkovská M. 1 

1Institute of Histology and Embryology, First Faculty of Medicine, Charles University in Prague, Czech Republic 

2Department of Obstetrics and Gynaecology, First Faculty of Medicine, Charles University and General University Hospital in Prague, Czech Republic 

8 Double immunohistochemical staining in routine diagnostic pathology 

Lodererova A., Honsova E., Voska L., Gabris V., Maluskova J. 

Department of Pathology, Institute for Clinical and Experimental Medicine, Prague, Czech Republic 

9 Micro RNA Assessment as a New Diagnostic and Prognostic Tool of Barrett´s Esophagus. Pilot Study 

Luzna P. 1 , Gregar J. 2 , Uberall I. 3 , Prochazka V. 2 , Ehrmann J. jr. 1,3 

1 Department of Histology and Embryology, Faculty of Medicine and Dentistry, Palacky University and University Hospital Olomouc, Czech Republic 

(pavla83@hotmail.com) 

2 nd II Internal Clinic, Faculty of Medicine and Dentistry, Palacky University and University Hospital Olomouc, Czech Republic 

3Laboratory of Molecular Pathology, Faculty of Medicine and Dentistry, Palacky University and University Hospital Olomouc, Czech Republic 

10 Prognostic factors in multiple myeloma 

Mareckova J. 1 ,Scudla V. 2 , Ehrmann J. 1 , Abrahamova P. 1 

1Department of Pathology, Laboratory of Molecular Pathology, Faculty of Medicine and Dentistry, Palacky University Olomouc, Czech Rebublic 

2 rd 3 Internal clinic, Faculty of Medicine and Dentistry, Palacky University Olomouc, University Hospital Olomouc, Czech Republic 

11 Determination of metallothioneins and alpha-methylacyl-CoA racemase in tumor prostate diseases 

Masarik M. 1 , Gumulec J. 1 , Sztalmachova M. 1,2 , Hlavna M. 1 , Kuchtickova S. 1 , Rovny A. 3 , Hrabec R. 3 , Eckschlager T. 4 , Krizkova S. 2 , 

Adam V. 2 , Kizek R. 2 

1Department of Pathological Physiology, Faculty of Medicine Masaryk University, Brno, Czech Republic 

2Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University, Brno, Czech Republic 

3Department of Urology, St. Anne´s University Hospital, Brno, Czech Republic 

4Department of Paediatric Haematology and Oncology, 2nd Faculty of Medicine Charles University in Prague, Czech Republic 

12 Identifi cation of grafted bone marrow cells in the recipient tissues 

Mokrý J. 1 , Filip S. 2 , Vávrová J. 3 , Čížková D. 1 , Šinkorová Z. 3 , Mičuda S. 4 

1Department of Histology and Embryology, Charles University in Prague, Faculty of Medicine, Hradec Králové, Czech Republic 

2Department of Oncology and Radiotherapy, Charles University in Prague, Faculty of Medicine and Teaching Hospital, Hradec Kralové, Czech Republic 

3 Department of Radiobiology, University of Defence Brno, Faculty of Military Health Sciences, Hradec Králové, Czech Republic 

4 Department of Pharmacology, Charles University in Prague, Faculty of Medicine, Hradec Králové, Czech Republic 


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PROGRAMME 

13 Role of apoptosis associated genes in predicting clinical outcome of breast cancer 

Skálová H. 1 , Dundr P. 1 , Povýšil C. 1 , Velenská Z. 1 , Berková A. 1 , Staněk L. 1 , Dlouhá Z. 2 , Petruželka L. 2 , Tvrdík D. 1 

1Institute of Pathology, 1st Faculty of Medicine, Charles University and General University Hospital in Prague, Czech Republic 

2Department of Oncology, 1st Faculty of Medicine, Charles University and General University Hospital in Prague, Czech Republic 

14 Decalcifi cation options of bone material for consequential molecular biological and cytogenetic diagnosis – 

from the experimental model to trepanobiopsy 

Staněk L. 1,2,3 , Tvrdík D. 1 , Střítský J. 1 , Lísová S. 1 , Berková A. 1 , Ludvíková M. 1,3 

1Institute of Pathology, General Faculty Hospital and 1st Faculty of Medicine, Charles University in Prague, Prague, Czech Republic 

2 Department of Experimental virology, IHBT, Prague, Czech Republic 

3 Institute of Biology, Faculty of Medicine in Pilsen, Charles University, Pilsen, Czech Republic 

15 Role of PPARγ in colon cancer cells 

Straková N. 1,2,3 , Hofmanová J. 1,3 , Tylichová Z. 1,3 , Knopfová L. 4 , Šmarda J. 4 , Kozubík A. 1,3 , Ehrmann J. 2,3 , Kolář Z. 2,3 

1Laboratory of Cytokinetics, Institute of Biophysics, Acad Sci Czech Rep, v.v.i., Brno, Czech Republic 

2Laboratory of Molecular Pathology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic 

3Biomedical Centre of Institute of Biophysics Acad Sci Czech Rep, v.v.i. and Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic 

4 Laboratory of Cell Diff erentiation, Institute of Experimental Biology, Masaryk University, Brno, Czech Republic 

16 Molecular genetics changes in melanocystic lesions – case report 

Uvírová M. 1 , Dvořáčková J. 2 , Šimová J. 1 , Urbanovská I. 1 , Žiak D. 1 , Konvalinka D. 1 , Kubová B. 1 

1CGB laboratory, Ostrava, Czech Republic 

2Faculty of Medicine University of Ostrava, Ostrava, Czech Republic 

17 Role of matrix metalloproteinase-19 in liver fi brosis 

Zbodakova O. 1 , Jirouskova M. 1 , Sedlacek R. 1 , Ehrmann J. 2 , Hajduch M. 3 , Jirkovska M. 4 

1Laboratory of Transgenic Models of Diseases, Institute of Molecular Genetics of the ASCR, Prague, Czech Republic 

2Department of Pathology, Medical Faculty, Palacky University, Olomouc, Czech Republic 

3Laboratory of Experimental Medicine, Department of Pediatrics and Oncology, Palacky University and University Hospital, Olomouc, Czech Republic 

4 Institute of Histology and Embryology, First Faculty of Medicine, Charles University, Prague, Czech Republic 

SOCIAL PROGRAMME 

Time 

17:30–18:00 Welcome drink 

18:00–19:00 Concert in the Olomouc Archidiocesan Museum 

19:00–19:30 Hot wine time 

20:00–24:00 Social evening in Archa restaurant – Svatý Kopeček (bus transport guaranteed) 

Workshop on Molecular Pathology 

SATURDAY, APRIL 30, 2011 

Time Chairs: M. Mistrík, I. Überall (Olomouc) Venue/language 

9:00–9:30 P. Krist (Prague) 

3D Microscopy 

9:30–10:00 M. Mistrík (Olomouc) 

Applicated fl uorescence microscopy 

10:00–10:20 Coff ee break 


Theoretical Institutes Building 

Faculty of Medicine and Dentistry, Palacký 

University Olomouc / Czech 

Theoretical Institutes Building 



10:20–11:40 3D Microscopy – Practical demonstration by Zeiss (part I) Theoretical Institutes Building 



11:40–13:00 3D Microscopy – Practical demonstration by Zeiss (part II) Theoretical Institutes Building 


University Olomouc / Czech

PROGRAMME 

96. olomoucký meziregionální mezioborový diagnostický seminář 

Společnosti českých patologů a české sekce 

International Academy of Pathology 

PÁTEK, 29. DUBNA 2011 

Čas Místo konání/jazyk 

8:30–9:00 Registrace 

9:00–9:15 Slavnostní zahájení Regionální centrum 

sál Centaurus / anglicky, česky 

Diagnostický seminář Společnosti českých patologů a české sekce International Academy of Pathology 

Předsednictvo: M. Tichý, M. Geierová (Olomouc) 

9:15–10:45 Diagnostický seminář – část I Regionální centrum 

sál Andromeda / česky 

10:45–11:15 J. Berounský (České Budějovice) 

Vakuový systém pro přepravu a uchování biologického materiálu 

11:15–11:45 Přestávka, občerstvení 

Téma: Tissue Banking 

Předsednictvo: P. De Blasio (Milano), L. Dušek (Brno) 



12:05–12:25 L. Dušek (Brno) 

How to manage real time data support for registries and tissue banking 

in clinical practice? Cancer care case study 



research 

Regionální centrum 



sál Centaurus / anglicky 





12:45–13:00 Diskuze Regionální centrum 


13:00–14:00 Oběd 

Sekce posterů 

14:00–16:00 Diagnostický seminář – část II Regionální centrum 


SPOLEČENSKÝ PROGRAM 

Čas 

17:30–18:00 Uvítací přípitek 

18:00–19:00 Koncert v Arcidiecézním muzeu v Olomouci 

19:00–19:30 Zahřátí u svařeného vína 

20:00–24:00 Společenský večer v restauraci Archa na Sv. Kopečku (odvoz autobusy zajištěn) 

Ve dnech 28. dubna – 1. května 2011 probíhá v Olomouci „Jarní Flora Olomouc“. Vřele doporučujeme! 


7

8 

PROGRAMME 

Workshop molekulární patologie 

SOBOTA, 30. DUBNA 2011 

Čas Předsednictvo: M. Mistrík, I. Überall (Olomouc) Místo konání/jazyk 

9:00–9:30 P. Krist (Praha) 

Mikroskopie v 3D 


Aplikovaná fl uorescenční mikroskopie 


10:20–11:40 Mikroskopie v 3D – praktická demonstrace I 

fi rmy Zeiss 

11:40–13:00 Mikroskopie v 3D – praktická demonstrace II 

fi rmy Zeiss 


Teoretické ústavy LF UP / česky 

Teoretické ústavy LF UP/ česky 


Teoretické ústavy LF UP / česky

3. olomoucké dny histologických laborantů 

PÁTEK, 29. DUBNA 2011 

PROGRAMME 

Čas Místo konání/jazyk 

8:30–9:00 Registrace 

9:00–9:15 Slavnostní zahájení Regionální centrum 


Konference histologických laborantů I 

Předsednictvo: J. Ehrmann, D. Kvapilová (Olomouc) 

9:15–9:35 L. Vodičková, A. Faltýnková., D. Novotná (Olomouc) 

Laboratorní zpracování neodvápněné kostní tkáně 

9:35–9:55 P. Skýpalová, M. Tichý, K. Žamboch (Olomouc) 

Využití histomorfometrie kostní dřeně k diagnostice renálních osteopatií 

9:55–10:15 J. Ehrmann (Olomouc) 

Koncept řešení histologických laboratoří – nové trendy 

10:15–10:35 J. Rázga (Brno) 

Produktivita práce v patologické laboratoři 

10:35–11:05 M. Richter (Praha) 

Automatizace imunohistochemie a in situ hybridizace systémy Ventana - 

nové metody a přístupy 

11:05–11:15 Diskuze 


Téma: Tissue Banking 

Předsednictvo: P. De Blasio (Milano), L. Dušek (Brno) 



12:05–12:25 L. Dušek (Brno) 

How to manage real time data support for registries and tissue banking in 

clinical practice? Cancer care case study 



research 


sál Centaurus / česky 















12:45–13:00 Discussion Regionální centrum 


13:00–14:00 Oběd 

Sekce posterů 

Konference histologických laborantů II 

Předsednictvo: D. Kvapilová, J. Ehrmann (Olomouc) 

14:00–14:20 P. Hajzlerová, J. Mokrý (Hradec Králové) 

Využití tyramidů v imunohistochemii 

14:20–14:40 P. Lužná (Olomouc) 

Laserová mikrodisekce 

14:40–15:10 E. Knopek (Praha) 

BRADY – Laboratorní značení 

15:10–15:30 M. Janíková, J. Škarda, V. Žižková (Olomouc) 

Využití parafínových bloků k izolaci miRNA 

15:30–15:50 M. Ondráková (Brno) 

Informace ČAZL 












9

10 

15:50–16:00 Diskuze 

POSTERY 

PROGRAMME 

1 Tkáňový procesor TISSUE PROCESSOR STP 420D 

Tylečková M., Muchová A. 

CGB laboratoř, Ostrava – Vítkovice, Česká republika 

2 Porovnání hladin PSA s histologickým gradingem karcinomu prostaty 

Němečková L. 1 , Hafuda A. 2 , Martinková L. 1 , Vodičková J. 1 , Hladíková L. 1 , Straka V. 1 ,Hornych J. 3 

1Oddělení patologie, Oblastní nemocnice Náchod, Česká republika 

2Oddělení urologie, Oblastní nemocnice Náchod, Česká republika 

3Oddělení klinické biochemie a diagnostiky, Oblastní nemocnice Náchod, Česká republika 

SPOLEČENSKÝ PROGRAM 

Čas 

17:30–18:00 Uvítací přípitek 

18:00–19:00 Koncert v Arcidiecézním muzeu v Olomouci 

19:00–19:30 Zahřátí u svařeného vína 

20:00–24:00 Společenský večer v restauraci Archa na Sv. Kopečku (odvoz autobusy zajištěn) 

Ve dnech 28. dubna – 1. května 2011 probíhá v Olomouci „Jarní Flora Olomouc“. Vřele doporučujeme! 

Workshop molekulární patologie 

SOBOTA, 30. DUBNA 2011 

Čas Předsednictvo: M. Mistrík, I. Überall (Olomouc) Místo konání / jazyk 

9:00–9:30 P. Krist (Praha) 

Mikroskopie v 3D 


Aplikovaná fl uorescenční mikroskopie 


10:20–11:40 Mikroskopie v 3D – praktická demonstrace I 

fi rmy Zeiss 

11:40–13:00 Mikroskopie v 3D – praktická demonstrace II 

fi rmy Zeiss 





Teoretické ústavy LF UP / česky

Honorary guests 

Juri Kopolovic, Professor, M.D., Ph.D. 

Juri Kopolovic is currently Professor of Pathology at the Hebrew University – Hadassah 

Medical Center in Jerusalem. He completed his medical studies in 1972 at the Hebrew 

University – Hadassah School of Medicine. During the residency at Hadassah Medical 

Center he became interested in nephropathology and gynecopathlogy. In 1986–1988 he 

continued his training with a clinical and research fellowship at the Pathology Department 

at the Beth Israel Hospital of the Harvard School of Medicine in Boston where he worked 

on morphological changes along the nephron in rat experimental model of hypoxia and 

reperfusion. Upon returning to Israel he continued in research in the field of autoimmunity, 

atherosclerosis and extracellular matrix in malignancies of the female genital tract. It was 

shown that in experimental model of mice SLE treatment with intravenous gamma globulin 

lads to a complete clinical and morphological remission of SLE. He studied the role 

of extracellular matrix in local invasion, metastasis angiogenesis and prognosis in female 

genital tract malignancies. He is presently the chairman of the Department of Pathology 

at the Hebrew University School of Medicine, Jerusalem, Israel. 

Jaroslav Mokrý, Professor, M.D., Ph.D. 

Jaroslav Mokrý currently holds the position of Head of the Department of Histology and 

Embryology at Charles University Medical Faculty in Hradec Kralove. He has been working 

in the Department of Histology and Embryology at Charles University Medical Faculty in 

Hradec Kralove since 1985. He completed his M.D. degree in general medicine from Charles 

University in Prague in 1990. In 1993–1994 he was on his research stay in the Department 

of Human Anatomy, Oxford University, UK, where he was engaged in cultivation of fetal 

dopaminergic neurons. He defended his Ph.D. thesis in 1995 and was appointed Professor 

of Histology and Embryology in 2006. He was awarded Charles University Rector’s Prizes in 

1987 and 1990 and Young Histochemist Award from International Federation of Societies 

of Histochemistry and Cytochemistry in Kyoto, Japan, in 1996. He is president of Czech 

Society for Histo- and Cytochemistry and vice-president of Czech Anatomical Society. He 

published 130 research articles and presented over 300 lectures or posters. Main focus 

of his scientific activity covers the areas of stem cell biology, regenerative medicine, cell 

cultivation, cell transplantation, cell therapy, cell differentiation, angiogenesis, detection 

of molecules in situ and histology. 

ABSTRACT BOOK 

CURRICULUM VITAE 


11

12 ABSTRACT BOOK 

Invited guests 

Petr Dubový, Professor, D.Sc., Ph.D. 

Petr Dubový currently holds the position of Professor and Head of the Department of 

Anatomy and Senior Researcher in the Central European Institute of Technology (CEITEC), 

Masaryk University (MU) in Brno. He obtained his Master and Doctoral degree of Biology 

in the Faculty of Natural Science and was subsequently graduated and then taken degree 

of Professor of Human Anatomy in the Medical Faculty of MU. He was visiting scientist in 

the Department of Anatomy and Neuroscience Karolinska Institutet, Stockholm (1991– 

1995). In the period of 2001–2009, he was chairman of the Czech Society for Histo- and 

Cytochemistry. He is interested predominantly in the cellular and molecular biology of 

neuron-glial cell interactions during degeneration and regeneration of the nervous system, 

especially using in situ proteomics (immunohistochemistry). His recent scientific work is 

directed to involvement of cytokines, chemokines and their receptors in neuropathic 

pain induction as simultaneous process of nerve regeneration. He has published over 100 

peer-reviewed papers as author and co-author. 


Expression of cytokine proteins and mRNAs in the primary sensory neurons 

of neuropathic pain model 

Petr Dubový 

Department of Anatomy, Faculty of Medicine, and Central European Institute of Technology (CEITEC), Masaryk University, Brno, 

Czech Republic 

Peripheral neuropathic pain, manifested 

as spontaneous pain, hyperalgesia and 

allodynia, arises as a result of many forms 

of nerve damage. Compelling evidence 

indicates that neuropathic pain induced 

by nerve injury is related to cellular and 

molecular changes in the dorsal root 

ganglia (DRG) containing bodies of the 

primary afferent neurons. It is assumed 

that pro- and anti-inflammatory cytokines 

may contribute to both the induction and 

maintenance of neuropathic pain. Chronic 

constriction injury (CCI) of the rat sciatic 

nerve is frequently used experimental 

model of neuropathic pain. 

Unilateral CCI of the rat sciatic nerve 

performed under aseptic conditions was 

used to study changes of TNFa, IL-6 and 

IL-10 proteins and mRNAs in L4-L5 and 

C7-C8 DRG removed from both sides of 

naïve (n=16), operated (n=64) and shamoperated 

(n=64) rats. Neuropathic pain 

induction was tested by withdrawal 

threshold of mechanoallodynia and 

thermal hyperalgesia. Levels of TNFa, IL- 

6 and IL-10 proteins and their mRNA were 

analyzed by quantitative immunohistochemistry, 

ELISA, Western blot, in situ 

hybridization and RT-PCR in DRG samples 

from naïve, CCI- and sham-operated rats 

for 1, 3, 7 and 14 days. 

The naïve DRG displayed no or very 

weak immunofluorescence staining for 

the cytokine proteins in the neuronal 

bodies with a moderate intensity in small 

and medium-sized neurons. Unilateral 

CCI induced bilateral elevation of TNFa, 

IL-6 and IL-10 immunostaining usually in 

large-sized neuronal perikarya of both 

L4-L5 and C7-C8 DRG. ELISA and Western 

blot confirmed bilateral and temporal increase 

of TNFa, IL-6 and IL-10 proteins not 

only in the homonymous lumbar DRG 

associated with the sciatic nerve, but 

also in the heteronymous cervical ones 

non-associated with spinal segments of 

constricted nerve. 

TNFa protein was peaked in L4-L5 

DRG at 7 and C7-C8 DRG at 14 days after 

CCI, while IL-6 protein level was the highest 

in both lumbar and cervical DRG at 3 

days. In contrast, IL-10 protein was significantly 

increased in lumbar and cervical 

DRG of both sides at 1 day and 3 days. 

Elevated levels of TNFa and IL-6 mRNAs 

were proved in the neuronal bodies of all 

DRG from CCI rats by in situ hybridization 

and verified by RT-PCR. 

Surprisingly, sham-operated rats displayed 

bilateral increased staining for IL-6 

mRNA in satellite glial cells of both cervical 

and lumbar DRG, while staining for 

TNFa mRNA was enhanced in neuronal 

bodies. These increased levels of mRNAs 

were confirmed by RT-PCR 

Our results indicate that neuroinflammation 

expressed by increased production 

of cytokines is not limited to the primary 

sensory neurons of injured nerve, 

but is spread to the contralateral side 

as well as alongside neuroaxis to DRG 

neurons of remote segments. In addition, 

neuroinflammatory reaction of satellite 

glial cells in sham-operated rats suggests 

their sensitivity to tissue injury. Both 

neuroinflammatory reactions influence 

excitation of the primary sensory neurons 

and may participate in neuropathic 

pain induction. Moreover, changes of 

cytokine proteins in DRG non-associated 

with damaged nerve could be related 

with a general neuroinflammatory reaction 

of the nervous system to injury and 

thereby promote potential of the DRG 

neurons for regeneration of their axons 

following a conditioning lesion. 

Supported by MSM0021622404.

Eli Pikarsky, M.D., Ph.D. 

Eli Pikarsky is at the Department of Pathology and the Lautenberg Center for Immunology. 

He received his M.D. and Ph.D. at the Hebrew University, Jerusalem, Israel in 1992 and 2000 

respectively. He actively participates in clinical duties at the department of Pathology and 

heads the laboratory for molecular pathology which he established. Dr. Pikarsky’s research 

career is devoted to studying tissue interactions which are important for cancer initiation, 

promotion and progression. His research group includes 2 research associates holding 

a Ph.D. degree, 6 doctoral students and several technicians. His studies involve analysis 

of the interaction between inflammation and cancer cells and elucidating the roles of 

several transcription factors in cancer biology. He is a member of the steering committee 

of the Israel National Tissue Bank. He has received several awards at the Hebrew University 

including awards for excellence in research, in clinical practice and in teaching. He has also 

received the Daniel G. Miller Research Career Development Award from the Israel Cancer 

Research Fund, The Sir Zelman Cowen prize for discovery in medical research and the Teva 

Founders award for young scientists. 

ABSTRACT BOOK 

Advances in molecular diagnostics of hepatocellular carcinoma 

Eli Pikarski 

Department of Pathology and the Lautenberg Center for Immunology and Cancer Research, Jerusalem, Israel 

Hepatocellular carcinoma (HCC) 

afflicts more than 560,000 people 

worldwide each year and has one 

of the worst 1-year survival rates of 

any cancer type. Currently, there are 

no predictive markers for finetuning 

treatment of patients with HCC. We 

found that a chromosomal region 

encompassing VEGF-A is amplified in 

a subset of HCCs in humans and mice. 

This subset is histologically, molecularly 

and biologically distinct. Moreover, 

human HCCs that carry a synthenic 

VEGF-A-amplicon, also display distinctive 

histological features. Expression of 

VEGF-A in mice amplicon bearing 

tumors is associated with increased 

blood-vessel and macrophage density, 

and enhanced microenvironment 

derived HGF expression, which can 

support the proliferation of the 

malignant cells. Accordingly, we show 

that these amplicon-positive tumors 

are uniquely sensitive to treatment 

with soluble VEGF-A receptor or the 

multi-kinase inhibitor Sorafenib, which 


is the first line treatment for advanced 

HCC. Our data indicates that cancer 

gene amplification may have cell-nonautonomous 

oncogenic effects via the 

tumor microenvironment. Moreover, the 

VEGF-A amplicon is a potential biomarker 

for tumor response to direct and indirect 

VEGF blockers. Our study underscores 

the potential for clinical translation of 

results obtained from mouse models 

of human cancer guided by cancer 

genome analysis. 


13

14 

ABSTRACT BOOK 

José Moreira, M.Sc., Ph.D. 

José Moreira obtained his MSc. degree in Biochemistry from the Institute of Biomedical 

Sciences Abel Salazar in Porto, and a Ph.D. in Molecular Biology at the Dept. of Genetics, 

University of Copenhagen in 1998. Following a two-year period as Assistant Research 

Professor at the Dept. of Genetics, José Moreira took up a position as scientist at Active 

Biotech Research AB, Lund, Sweden. In 2002, he was appointed a senior scientist at the Dept. 

of Proteomics in Cancer, Danish Cancer Society. José Moreira is a member of the editorial 

board of Molecular and Cellular Proteomics and editorial manager for Molecular Oncology. 

His major achievements include contributions to some of the fundamental mechanisms 

underlying epigenetic regulation of genes, such as ATP-driven remodeling complexes being 

involved in activation as well as gene repression and the role that non-histone chromatin 

associated proteins play in the regulation of target genes. More recently, the identification 

of several biomarkers for bladder and breast cancer, as well as the establishment of 

a framework for biomarker discovery based on a new source, tissue interstitial fluid, 

and the identification a number of novel molecular markers that phenotypically define 

subpopulations of cells present in normal breast tissue. His current research interests 

focus on early detection and personalized treatment of cancer, in particular the concept 

of profiling of therapy-relevant protein markers in patients’ tumors and the development 

of a framework to identify biomarkers for blood-based diagnostic systems. 



Development of novel early detection and stratifi cation markers for breast 

cancer: integrating pathology and proteomics 

José Moreira 

Department of Genetics, Institute of Molecular Biology, University of Copenhagen, Denmark 

Our limited understanding of the biological 

impact of the whole spectrum of 

early breast lesions together with a lack 

of accurate molecular-based risk criteria 

for the diagnosis and assignment of 

prognostic significance to biopsy findings 

presents an important problem 

in the clinical management of patients 

harboring precancerous breast lesions. 

As a result, there is a need to identify biomarkers 

that can better determine the 

outcome of early breast lesions by identifying 

subpopulations of cells in breast 

premalignant disease that are at high-risk 

of progression to invasive disease. A first 

step towards achieving this goal will be 

to define the molecular phenotypes of 

the various cell types and precursors - 

generated by the stem cell hierarchy - 

that are present in normal, benign, and 

malignant conditions of the breast. For 

the past years our laboratory has carried 

out a systematic and comprehensive 

proteomic profiling of normal and malignant 

breast tissue in high-risk patients in 

a search for differentially expressed markers 

for early detection and stratification 

of patients, as well as novel targets for 

therapeutic intervention in breast cancer. 

This lecture will describe our efforts to 

address some of the issues that clinical 

proteomics is faced with, partly due to 

the formidable heterogeneity of tumors, 

but also due to limitations of the current 

proteomic technologies. The examples 

presented will include results from our 

work to identify proteins that characterize 

specific steps in the progression from 

early benign lesions to malignancy in 

breast apocrine carcinoma, the creation 

of a comprehensive database of proteins 

secreted/shedded by tumor cells and 

present in tissue interstitial fluid and from 

a systematic proteomic study to characterize 

the phenotypes of the different 

cell subpopulations present in normal 

human mammary tissue that can help 

define the molecular phenotypes underlying 

mammary epithelial normalcy. 

Finally results from our recent work on 

therapy-relevant stratification of triple 

negative breast cancer patients will also 

be discussed.

Ladislav Havel, Professor, D.Sc., Ph.D. 

Ladislav Havel obtained his academic degree (RNDr.) from Masaryk University in Brno in 1977, 

and Ph.D. at the Institute of experimental Botany, Academy of Sciences of the Czech Republic, 

Prague in 1983. He then worked at the same Institute, working place in Olomouc. In 1988 he 

moved to Mendel University in Brno. He became associate professor in 1988 and full time 

professor in 1998. Ladislav Havel is the head of the Department of Plant Biology at the Faculty 

of Agronomy (since 2007). He is a member of several international and national scientific 

societies. He published over 100 scientific communications in international scientific journals 

and monographs. He has been focused on the regeneration processes in plant tissue cultures 

of several species, especially somatic embryogenesis, the role of programmed cell death in 

plant development and in responses of plant cells to environmental stimuli. 

ABSTRACT BOOK 


Histochemistry in plant development and abiotic and biotic reactions 

Ladislav Havel 

Department of Plant Biology, Faculty of Agronomy, Mendel University of Agriculture and Forestry, Brno, Czech Republic 

Plant organisms have common characteristic 

of all eukaryotic organisms. In 

same features are different from other 

eukaryotes especially from animals and 

human, e.g. plant cells have cell wall and 

plastids, a special group of organelles. As 

in animals and human the histo- and/ 

or cytochemical methods are powerful 

tool in research of plant growth and development 

and responses to biotic and 

abiotic stimuli. Modern methods used 

in plant biology are usually optimized 

or derived from methods developed 

in animal and human research. A short 

survey of exploitation of these methods 

will be shown. 

15

16 

ABSTRACT BOOK 

Pasquale De Blasio, Professor, M.Eng. 

Pasquale De Blasio, is presently the CEO of BioRep (www.biorep.it), a global service provider 

biorepository based in Milan, Italy; President of ESBB (European, Middle-Eastern and African 

Society of Biopreservation and Biobanking, www.ESBB.org) and Adjunct Associate Professor 

at Temple University, Philadelphia-PA (USA) and founder of Integrated Systems Engineering 

S.r.l. (www.isenet.it) operating in development of automation instruments for molecular 

and cellular biology (e.g. Tissue. Microarrayers, Biorepositories automation, etc.). Pasquale De 

Blasio, has more than 35 years of industrial working experience in management positions. 

He worked for more than 14 years in Industrial and Power Plant design, and over 15 years in 

analytical instrumentation developments. He has also matured specific skills in multinational 

analytical instrumentation companies, as R&D Manager for the developments of HPLC 

Systems, UV/VIS Spectrophotometers, Ultra Centrifuges and Medical Anaesthesia Systems; 

and as Industrial Operations Management in the development and manufacturing of Blood 

Gas and Coagulation Analytical Instruments.. He has also experience with ISO 9001 Quality 

System, EEC Medical Directive and CE certification. Pasquale De Blasio, has a Bachelor of 

Science in Electrical Engineering from Villanova University, Villanova, Pennsylvania, USA 

(1976), and a Master Degree in Business Administration from Bocconi University, Milan, 

Italy (1986). He is co-author of several peer-reviewed scientific papers and co-participants 

of several industrial patents. 


De Blasio P. 1,2 and Biunno I. 2,3 

1BioRep Via Fantoli 16/15, Milano, Italy 

2Integrated Systems Egineering (ISE) Via Fantoli 16/15, Milano, Italy 

3Institute for Biomedical Research (ITB-CNR) Via F.lli Cervi 93, Milano, Italy 

“Tissue Banking” is an emerging 

concept with solid basis in translational 

medicine and in genetic epidemiology 

since allows biomedical research to 

flourish in several aspects: a tool for (i) 

biomarker identification, (ii) biomarker 

validation and (iii) subsequent drug discovery. 

Tissue banking aims to collect 

large number of high quality of samples 

process and store them in a manner that 

is compatible with the “omics” analysis. 

“Tissue microarray” have become an 

essential pathology tool being able to 

validate novel biomarkers in epidemiology 

scale since hundreds of tissue fragments 

can be analysed simultaneously. 


The advancement in digital pathology 

with whole-slide imaging and new techniques 

in image analysis and image databases 

has driven improvements in tissue 

samples collection and Tissue Banking. 

The identification of biomarkers is essential 

not only for diagnostic purposes 

but also as predictors of disease response 

to new and old drugs. This necessitates 

the use of tissue samples collected and 

stored in several centres, but in order to 

guarantee reproducible and comparable 

results, the multi-centre network must 

use standardized protocols (SOPs) and 

common minimum dataset of the clinical 

data. 


BioRep is a “Global Biorepository 

Service Provider” (www.biorep.it), which 

has implemented several Standard 

Operating Procedures (SOPs) developed 

thanks to its participation in several national 

and international research projects. 

BioRep has available SOPs for: tissues 

procurements and management, tissue 

microarrays, tracking system (strong IT 

structure), biosafety requirements, storage 

and distribution. 

It has assessed a code of conduct 

for the social, ethical and legal considerations.

Ladislav Dušek, Associate Professor, D.Sc., Ph.D. 

Ladislav Dušek (born 1967) is a director of Institute of Biostatistics and Analyses of Masaryk 

University Brno, Czech Republic. He graduated as an experimental biologist in Masaryk 

University in 1991, his further academic degrees obtained in fysiology of animals and in 

environmental sciences. Since 1994, he has been teaching at the Faculty of Science and 

Faculty of Medicine of Masaryk University. In 2004, he established a new research institute 

focused on biostatistics and health care informatics (IBA MU, www.iba.muni.cz) and since 

2004, he is a director of IBA MU. He is a chairman of coordination board of e-learning 

network of Czech and Slovak medical faculties MEFANET (www.mefanet.cz). He is a leader 

of more than 20 national cancer research programmes, focused namely on cancer screening 

(www.mamo.cz, www.kolorektum.cz, www.cervix.cz), epidemiology (www.svod.cz, www. 

uroweb.cz) and prospective registration of clinical data (www.registry.cz). He has published 

3 monographs, 5 authorized software tools in the field of cancer research and e-learning 

and more than 180 articles in international journals as an author or co-author. His research 

activities are focused on biostatistics, application of statistical methods in biology and in 

clinical sciences, health care informatics, cancer epidemiology and biology, human and 

ecological risk assessment. 

ABSTRACT BOOK 


How to manage real time data support for registries and tissue banking in clinical 

practice? Cancer care case study 

Ladislav Dušek 

Institute of Biostatistics and Analyses of Masaryk University Brno, Czech Republic 

In the era of personalized medicine, 

detailed description of the disease has 

become of the same importance as identification 

of a patient. It is particularly the 

case of chronic illnesses, which require 

multidimensional scoring of their stage 

and risk status. Personalized medicine 

strongly increases the value of tissue 

banking because we can turn back in 

time searching for useful prognostic 

factors or performing land mark analyses 

of therapeutic response or survival. 

However, with the current dynamic arrival 

of many new biomarkers, corresponding 

research activity should address how 

to link the new dimensions with standard 

clinical data in order to utilize their incremental 

value to existing diagnostic 

methods. Moreover, targeted treatment 

of many chronic diseases (e.g. breast 

cancer, chronic leukaemia) brings a new 

challenge for attempts to measure the 

therapeutic results, as the patients can 

experience multiple disease-free periods 

during the course of the therapy. The 

added value of the therapeutic progress 

depends on whether we can link common 

clinical data with new molecular 

and genetic markers which are able to 

cope with multiple disease remissions 

or progressions in time. 

Any banking of clinical material 

should be coupled with complete diagnostic 

and therapeutic identification of 

recorded subjects otherwise the information 

stored is insufficient for future 

views. Comprehensive electronic health 

records (EHR) stand in the position of 

key factor limiting current progress in 

therapeutic optimization. There are two 

principal ways how to manage support 

for personalized medicine, focus on clinical 

registries or automated collection of 

data from hospital information systems 

(HIS). This presentation proposes a novel 

approach combining both these data 

sources in hospital–based data mining 

system supporting analyses of cancer 

data and tissue banking. 

Common HIS is composed of a number 

of modules and satellite systems 

producing large volumes of heterogeneous 

data. Most of the outcomes are 

produced to fulfill administrative duties, 

namely reporting for health care payers. 

In such administrative outcomes, 

the clinical information is inevitably insufficient 

and many clinically important 

views are not possible. It limits all fields 

of medicine, particularly the disciplines 

which require prognostic typology of patients, 

based not only on diagnosis itself. 

Cancer care can serve as a typical model 

of such information demanding field because 

in most of cancers, the therapeutic 

standards are related to diagnosis, clinical 

stage, grade and other prognostic 

markers. These attributes are however 

not well standardized in HIS data capture 

systems. Their retrospective availability 

can be managed only through 

external sources like population-based 

registries. 

This study documents functionality of 

a newly developed system which is able 

to merge records of the Czech National 

Cancer Registry (CNCR) and hospital administrative 

data related to cancer care. A 

new data mining tool provides solution 

for this highly required data merge. The 

proposed system is capable to generate 

synthesis of different data sources 

and to extract information relevant for 

the evaluation of cancer care. Database 

of CNCR contains of more than 1.6 million 

of recorded cancer cases collected 

in a standardized way in past 30 years. 


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18 

ABSTRACT BOOK 

In combination with HIS data from any 

given hospital, the CNCR is able to map 

epidemiologic load and to identify hospital 

case mix in this segment of medical 

care. The CNCR thus supports the weakest 

point in parametric data collection, 

i.e. retrospectively available, fully representative 

diagnostic typology of cases. 

Successful implementation of the 

proposed system will be demonstrated 

over data from five cancer centers in the 

Czech Republic. The final merged database 

consists of more than 500 thousands 

of cancer care records containing 

all necessary diagnostic items and records 

on all therapeutic steps and corre- 


sponding outcomes. Available follow-up 

period exceeds 5 years in each participating 

hospital. The system can be easily 

linked to any tissue bank, registry or 

individually-based set of laboratory data. 

Comprehensive parametric structure 

forms a framework enabling all necessary 

views in the data, e.g. sorting according 

to demography, diagnoses, tumor morphology, 

or TNM classification of tumors. 

Routinely generated hospital records 

further contribute to the identification 

of therapeutic episodes, their content, 

cost and outcomes. Any examination 

of a biomarker is therefore associated 

with exactly measured disease status at 

a given time and its value is attributed to 

relevant therapeutic steps. The presentation 

introduces a new version of hospital 

reporting system which enables on-line 

analyses of the merged data 

Acknowledgements: Validation of 

the Czech National Cancer Registry 

and population–based monitoring of 

cancer disparities are supported by 

grant “Addressing Cancer Disparities 

in Central and Eastern Europe” Bristol- 

Myers Squibb Foundation, 2009 – 2011 

(Project: National Information System for 

the Assessment and Communication of 

Cancer Care Results and Quality in the 

Czech Republic).

Giorgio Stanta, Professor, M.D., Ph.D. 

Giorgio Stanta is Professor of Pathology at the Medicine, Surgery and Health Department at 

the University of Trieste. His main interest is molecular medicine and particular translational 

and diagnostic molecular pathology, mainly oncology. He is the head of the Molecular 

Histopathology Laboratory in the Cattinara Hospital in Trieste. He participates, as an 

expert, in the Biobanking and Biomolecular Resources Research Infrastructure (BBMRI). 

He is the coordinator of the European Group for molecular analysis in archive tissues of 

the European Society of Pathology and he is a member of the Management Board of 

“Molecular Pathology Study Group” of “European Society of Pathology” and a member of 

the executive committee of the Italian Reference Centre for BBMRI (European Infrastructure 

for Biobanking) as an expert in molecular analysis in human tissues and tissue biobanks. 

Giorgio Stanta is the coordinator of the European group IMPACTS “Archive tissues: improving 

molecular medicine research and clinical practice”, involving around twenty major university 

hospitals in 11 countries, with over 100 researchers (www.impactsnetwork.eu) and the 

coordinator of the biobanking network of pathological tissues at the Italian national level 

(NIPAB – Network of Italian Pathology Archives Biobank) and he is also the coordinator of 

the European biobanking network “Pan-European Archive Tissue Biobanking Network”, 

involving European pathologists in collaboration with the European Society of Pathology 

(ESP) and with the European Infrastructure for biobanks (BBMRI). He is the editor of the 

“Guidelines for Molecular Analysis in Archive Tissues”, those tissues which are fixed and 

paraffin embedded (paraffin is the only clinical material used in hospital for any kind of 

diagnostics). This book will be soon published by Springer Verlag. Giorgio Stanta was 

appointed as an expert by the European Commission for FP7. 

Archive tissue biobanking as a basis for translational 

and reverse translational research 

ABSTRACT BOOK 

Giorgio Stanta 

Department of Medical, Surgical and Health Sciences, University of Trieste, Italy, stanta@impactsnetwork.eu 

Fixed and paraffin-embedded tissues 

(Archive Tissues – AT) are the only 

tissues usually available for any patient. 

For this reason any diagnostic molecular 

analysis should be performed in this 

type of tissues. Nowadays we are able 

to perform any kind of nucleic acid or 

proteomics analysis in AT and most of 

these methods have been validated. 

Preservation of tissues can be quite 

different depending on pre-analytical 

conditions and special effort should be 

done to standardize the treatments. 

AT is the only available collection of 

tissues that really represent the clinical 

heterogeneity of human diseases. In 

fact, we have in our pathology archives 

any kind of even rare lesions, with also 

well defined subtypes, clinical information 

and long follow-up periods. 


Pathologists are the only experts able to 

correctly microdissect tissues to obtain 

reliable and reproducible analytical results. 

Moreover, they have a central position 

in the harmonization of the huge 

morphological-histological knowledge, 

reached in human pathology with the 

new molecular information. 


19

20 

ABSTRACT BOOK 

Paul G. Murray, Professor, M.Sc., Ph.D. 

Paul G. Murray is currently Professor of Molecular Pathology at the School of Cancer 

Sciences of the College of Medical and Dental Studies, University of Birmingham. The major 

focus of his research group is to provide a better understanding of the molecular events 

leading to the development of Hodgkin´s lymphoma and especially to study the role of 

the Epstein-Barr virus in this process. He has succesfully finished 38 research grants and 

published 78 original research papers and 14 review articles since 1989. He also published 

7 books and 3 textbooks. Paul G. Murray is a regular reviewer for over 20 peer-reviewed 

journals including Blood, American Journal of Pathology, Journal of Pathology, Oncogene 

and Cancer Research. Last but not least, he has succesfully supervised many Ph.D. students, 

presented multiple invited lecture at international conferences and is a member of several 

editorial boards and scientific societies. 

Small lipids as therapeutic targets in cancer 



Murray P.G. 1,2 , Miscoria M. 1 , Abdullah M. 1 , Yap L.F. 1 

1School of Cancer Sciences, University of Birmingham,Birmingham, United Kingdom 

2 Laboratory of Molecular Pathology & Department of Pathology and Institute of Molecular and Translation Medicine, 

Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic 

Two related oncogenic phospholipids, 

sphingosine-1-phosphate (S1P) and 

lysophosphatidic acid (LPA) are implicated 

in the pathogenesis of variety of 

cancer types. We have identified that 

these lipids work at multiple levels in 

the pathogenesis and maintenance of 

the tumour phenotype of different cancer 

types. Work presented here demonstrates 

the therapeutic potential of 

targeting these lipids and the signaling 

pathways they regulate in the treatment 

of several malignancies with poor prognosis, 

including those of the head and 

neck cancer and kidney.

Karel Smetana, Professor, M.D., Ph.D. 

Karel Smetana is Professor of Anatomy at the Institute of Anatomy, 1 st Faculty of Medicine, 

Charles University, from where he graduated in 1983. Dr. Smetana is a member of many 

professional commissions and scientific societies, including the Grant Agency of the Czech 

Republic. His work is focussed on cell and developmental biology, especially on squamous 

epithelia in physiological and pathological condition. He is an author of more than 170 

papers, patents and textbook chapters, with articles cited more than 750 times and an 

H-index of 19. His work has been recognized by the National Scientific Award of the Czech 

Republic (2002), by award of Int. J. Oncol. (2005) and by the Annual Award of Minister of 

Education, Youth and Sport of the Czech Republic (2010). 

Social life of cancer cell 

ABSTRACT BOOK 


Smetana K., Jr., Dvořánková B., Lacina L., Szabo P., Kolář M., Strnad H. 

Charles University, 1 st and 2 nd Faculty of Medicicine, Institute of Anatomy and Center of Cell Therapy and Tissue Repair, 

Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic; karel.smetana@lf1.cuni.cz 

Increasing volume of data demonstrate 

important role cancer stem cells in 

tumor formation and spreading through 

organism. Because, the biology of normal 

tissue stem cells including the maintenance 

of their stemness is influenced 

by the surrounding microenvironment, 

the role of tumor microenvironment for 

its biology is also extensively studied. 

Based on results of many laboratories, 

the cancer stem cell niche has been 

placed to the tumor stroma. This tissue 

is rather heterogenic and it is composed 

from fibroblasts producing extraxcellular 

matrix, inflammatory cells and from the 

capillaries. The extensive crosstalk between 

stromal elements and cancer cells 

including stem cell pool is expectable. 

Among the stromal cells, the stromal 

fibroblasts seem to be of a great importance. 

Although, their origin is not clear, 

they are biologically active and they are 

able to influence the behavior of both 

the normal and cancer epithelial cells. 

The differences between both the normal 

and cancer stromal fibroblats were 

characterized using the cell biology and 

molecular genetics microarray approach. 

The obtained data indicate the importance 

of epithelial-mesenchymal interaction 

in tumor biology with therapeutic 

perspective in future. 


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22 

Jakub Sikora, M.D., Ph.D. 

ABSTRACT BOOK 

Jakub Sikora graduated in 2000 and received Ph.D. in cell biology and pathology at the 

Charles University in Prague in 2007. Dr. Sikora is an assistant professor (lectures cell biology 

and cell pathology) at the Institute of Inherited Metabolic Disorders (1 st Faculty of Medicine, 

Charles University in Prague) where he heads the Laboratory of advanced microscopic 

techniques and tissue cultures, his professional interests include pathology of inborn errors 

of metabolism with special focus on lysosomal storage/dysfunction disorders. For his 

accomplishements, Dr. Sikora received several academic awards such as Prize of Professor 

Karel Weigner or Prize of Hlavka´s Foundation. 



Danon disease – lysosomal dyfunction disease – molecular pathology 

of LAMP2 protein – implications for clinical diagnostics 

Sikora J. 1 , Majer F. 1 , Kubánek M. 2 , Vlášková H. 1 , Stolnaya L. 1 , Kró l. 3 , Kalina T. 3 , Dvořáková L. 1 and Elleder M. 1 

1 st Institute of Inherited Metabolic Disorders, Charles University in Prague – 1 Faculty of Medicine, Czech Republic 

2Cardiology Department, Institute for Clinical and Experimental Medicine, Prague, Czech Republic 

3 Department of Paediatric Haematology and Oncology, Childhood Leukaemia Investigation Prague, 

Charles University in Prague – 2nd Faculty of Medicine, Czech Republic 

Danon disease (DD) is a monogenic 

X-linked disorder, which is clinically characterized 

by combination of cardiomyopathy, 

skeletal myopathy and variable 

degree of mental retardation. DD develops 

due to mutations in the gene coding 

lysosomal associated membrane protein 

2 (LAMP2). The ultrastructural changes in 

DD reflect a defect in autophagosomal 

(AP) clearance due to inefficient AP/lysosomal 

membrane interaction and fusion 

impaired by defects in LAMP2. 

We present a DD family with a clinical 

phenotype of hypertrophic cardiomyopathy 

associated with pre-excitation, 

myopia and mild myopathy in affected 

males. The diagnosis of DD in the family 

members was based on the assessment 

of their clinical status and evaluation of 

the skeletal or cardiac muscle biopsies 

documenting typical histological and 

ultrastructural DD changes including 

the absence of LAMP2 protein by immuno 

staining. Direct sequence analysis 

of LAMP2 gene and its mRNA revealed 

a novel protein truncation mutation in 

all affected family members. 

Our major aim is to present the results 

of the studies on the cellular pathology in 

two explanted DD hearts, comment on 

the utility of peripheral white blood cells 

(WBCs) for clinical DD diagnostics, and 

discuss the functional studies focusing 

on the cellular processing of the novel 

pathologic LAMP2 protein variant. 

Histological findings corresponding 

to AP accumulation were present 

in all segments of the male DD 

hemizygote´s heart. By ultrastructural 

analysis, we have documented dilatations 

of T-tubular cardiomyocyte conductance 

system. Such changes, possibly 

affecting cardiomyocyte excitability 

and contractility, have never been, to 

our best knowledge, reported neither in 

endomyocardial DD biopsies or DD KO 

mice. Due to X chromosome inactivation, 

myocardium from a female DD heterozygote 

showed minute fraction of LAMP2 

immuno positive cardiomyocytes. 

To test the qualitative utility of WBCs 

in DD clinical diagnostics, we performed 

LAMP2 immuno detection in peripheral 

blood smears of all three patients. 

Samples from male hemizygotes (patient 

1 and 3) were devoid of LAMP2, whereas 

small fraction of myeloid lineage cells 

was LAMP2 positive in the female DD 

heterozygote. Electron microscopy of 

WBC pellets demonstrated scattered APs 

in neutrophilic granulocytes and monocytes, 

comparable to autophagic changes 

found in cardiomyocytes. LAMP2 

expression was further quantitatively 

evaluated in specific WBC sub-populations 

(granulocytes, CD4+ and CD8+ 

T-lymphocytes, CD20+ B-lymphocytes, 

CD14+ monocytes and CD56+ natural 

killer cells) by polychromatic flow cytometry 

analysis. This approach delineated 

granulocytes and CD14+ monocytes 

as the dominant LAMP2 expressing 

WBC types in healthy human controls. 

Whereas male DD patients lacked LAMP2 

in all WBC subtypes, female DD heterozygote 

expressed LAMP2 in 14 % of her 

peripheral granulocytes and monocytes. 

Comparative plotting of ratio values of 

LAMP2 expression in CD14+ monocytes 

over LAMP2 expression in CD20+ B lymphocytes 

provided a mean to clearly

discriminate DD patients from healthy 

subjects. Summarized, we propose to 

consider peripheral WBCs as optimal 

“first choice” cellular population for 

LAMP2 expression testing when DD is 

a clinically relevant differential diagnostic 

option. Flow cytometry analysis, in contrast 

to peripheral blood smear LAMP2 

labeling, represents a quantitative way 

to disclose a skewed LAMP2 expression 

in DD female heterozygotes 

As the novel mutation results in 

a putative truncated 334 amino acids 

long LAMP2 protein, which lacks the 

transmembrane and C-terminal cytosolic 

domain, we hypothesized that this 

mutated LAMP2 variant may attain a relatively 

stable folding state and while not 

targeted to late endosomes/lysosomes, 

may become exocytosed. Therefore we 

performed LAMP2 Western blotting of 

cardiac muscle, WBCs and cultured skin 

fibroblasts as well as corresponding extra 

cellular media (plasma and tissue culture 

medium) but found no intra- or extra 

cellular truncated LAMP2. Additionally, 

we compared the expression and intracellular 

targeting of the three physiological 

LAMP2 splicing variants (A, B and C) 

C-terminally tagged by GFP with a mutated 

lAMP2 fusion protein in cultured skin 

fibroblasts. Contrary to normal LAMP2 

ABSTRACT BOOK 

variants, which became expressed and 

were targeted to late endosomes/lysosomes 

as evaluated by GFP fluorescence, 

the mutated LAMP2 variant has not even 

been expressed (demonstrated by the 

lack of mature GFP fluorescence) suggesting 

folding instability and possibly 

its co- or early post-translational degradation. 

Acknowledgements: Malušková J., 

Honsová E. (Clinical and Transplantation 

Pathology Department – Institute for 

Clinical and Experimental Medicine, 

Prague, Czech Republic). 


23

24 

Rupert C. Ecker, Ph.D. 

ABSTRACT BOOK 

Rupert Ecker has been co-founder of TissueGnostics Austria and Chief Executive Officer. He 

is presently co-managing director in TissueGnostics-Romania and has held the position of 

a Chief Technology Officer. Before founding TissueGnostics he was a research scientist at the 

Competence Centre for Bio Molecular Therapeutics in Vienna, a joint venture between the 

University of Vienna and the Novartis Research Centre.As a co-inventor of the TissueFAXS 

technology Rupert Ecker has always been significantly involved in product development 

from system design to clinical testing and has successfully headed several joint R&D projects 

with academic partner institutions in the fields of advanced computer vision, cancer 

research, and stem cell biology. In March 2011 Rupert Ecker received the “Science2Business” 

award of the Austrian Chamber of Commerce for successfully managing joint industry & 

academia development collaborations.Rupert Ecker graduated in Cell Biology from the 

University of Vienna, doing his thesis at the research laboratories of the Depart. of Urology, 

General Hospital of Vienna. He has 17 years experience in microscopy and image analysis 

and, in addition, he has been trained in software development. 

Applications of Slide-Based Single Cell Cytometry by TissueFAXS 

in Histopathology 

Ecker R. C., Rogojanu R., Steiner G. 

TissueGnostics GmbH, Vienna, Austria 

Background: Automated identification 

and cytometric quantification of 

molecular markers (antibodies, histological 

stains, genetic probes) on the single 

cell level in tissue sections by means of 

slidebased cytometry has become an 

essential tool in biomedical research 

and routine analysis in histopathology. 

TissueFAXS is the microscopic equivalent 

to flow cytometry – applicable to tissue 

sections, cell culture monolayers and 

cell smears/cytospin preparations and 

allows observer independent analysis 

of all types of cells in all kinds of tissue 

sections in-situ. 

Technology & Method: The 

TissueFAXS system consists of a highend 

motorized fluorescence microscope 

and sophisticated software for image 

analysis. The samples can be stained by 

means of immunohistochemistry or immunofluorescence. 

By using the nuclear 

marker (e.g. hematoxilin or DAPI) the location 

of the individual cells is identified 


and the systém quantifies the staining 

intensity in each color channel for each 

and every cell. It allows even to distinguish 

between and compare between 

nuclear and cytoplasmatic marker expression 

in the same sample. For each 

cell or cellular compartment up to 14 

parameters are measured and stored in 

a database. Representation of cellular 

parameters may be obtained by dot 

plots and/or histograms in a FACS-like 

manner. 

Results: Results presented come from 

several studies and comprise different 

applications of slidebased cytometry. 

(i) Phenotypic characterization of tissue 

infiltrating leukocytes in tumor biology1, 

transplantation immunology2 and autoimmune 

diseases3 exemplifying the 

quantification of markers CD1a, CD3, 

CD4, CD8, CD11c, and CD68 in renal cell 

carcinoma, renal allograft rejection and 

atopic dermatitis. (ii) Signal transduction 

research in prostate cancer4,5 and brain 


neuropathology6 dealing with signal 

transducers and activators of transcription 

(STAT), and transforming growth 

factor beta. (iii) Quantification of proliferation 

marker Ki67 and Her2 in breast 

cancer. 

Discussion: The data above demonstrate 

novel approaches in research and 

improved data transparency in diagnosis 

using slide-based cytometry. Observerbiased 

visual estimation in immunohistological 

analysis of tissue samples is 

replaced by observer independent measurements 

on the single-cell level, which 

is especially important in multi-center 

studies and set-up of clinical studies for 

the pharmaceutical industry. 

1. Steiner GE, et al. J Immunol Methods 2000; 237(1– 

2): 39–35. 

2. Kozakowski N, et al. Nephrology Dialysis Transplantation 

2009; 24(6): 1979–1986. 

3. Bangert C, et al. Dermatology, 2010; 222(1): 36–48. 

4. Neuwirt H, et al. Am J Pathol 2009; 174(5): 1921–1930. 

5. Puhr M, et al. Cancer Res 2009; 69(18): 7375–7384. 

6. Lohr J, et al. Clinical Cancer Research, in press, 2011.

Increased susceptibility of MMP19 

defi cient mice to DSS-induced 

colitis provides novel insights 

into pathogenesis of intestinal 

infl ammation 

Brauer R. 1 , Dziechciarkova M. 2 , Skarda J. 3 , Hajduch M. 2 , 

Sedlacek R. 1 

1 Department of Transgenic Models of Diseases, Institute 

of Molecular Genetics AS CR, v.v.i, Prague, Czech Republic 

2 Laboratory of Experimental Medicine, Department 

of Pediatrics and Oncology, Medical Faculty, Palacky 

University, Olomouc, Czech Republic 

3 Department of Pathology, Medical Faculty, Palacky University, 

Olomouc, Czech Republic 

Introduction: Matrix metalloproteinase-19 (MMP19) belongs 

to a family of the zinc-dependent endopeptidases, which are 

known to remodel extracellular matrix (ECM) and process 

a number of cell surface receptors, their ligands, and adhesion 

molecules. These proteolytical activities have direct impact on 

cell behaviour, proliferation, and survival. Beside their role in 

matrix processing MMPs are important in the development and 

outcome of inflammatory responses by activating and releasing 

cytokines, establishing a chemokine gradient, controlling cell 

migration and survival, and contributing to epidermal barrier 

function. MMP19 was found to be constitutively secreted by 

various tissues and cell types and shown to be upregulated in 

skin under inflammatory conditions and during cancer progression. 

It was also shown that MMP19 controls delayed-type of 

hypersensitivity in the skin. Dependent on the cell and tissue 

type MMP19 can play either a suppressor role or can exacerbate 

the disease outcome. 

Aim: In this study, we aimed to analyse the functional role 

of MMP19 in intestinal inflammation by using MMP19-deficient 

mice in a mouse model of chemically-induced colitis. 

Material and methods: Dextran sodium sulphate (DSS) was 

administered to age-matched male wild-type (WT) and MMP- 

19 deficient mice (C57Bl/6 background) at a concentration of 

2 % (w/v) in drinking water for 6 days to induce acute colitis. 

For analysing the healing process mice were given 2 % DSS 

for 5 days followed by a 10 day recovery period using normal 

drinking water. The administration of two cycles of DSS (2 % 

DSS for 4 days, 10 days water) results in chronic colitis. Colitis 

severity was assessed by changes in body weight, stool consistency 

and occult bleeding. Furthermore, colon length was 

determined and histological analysis of its proximal, middle and 

distal parts was performed. Cytokine concentrations (IL-6, IL-1β, 

KC, MCP-1) in plasma and supernatants of colon tissue explants 

were measured by Luminex. 

Results: During the course of DSS-induced acute colitis, the 

MMP19-deficient mice lost significantly more weight than the 

WT mice (22 % ± 5 % vs. 12 % ± 3 %, p < 0.08). Also, the disease 

activity index (DAI) that included also the severity of diarrhoea 

and the amount of blood in stool was remarkably higher in 

ABSTRACT BOOK 

MMP19-deficient mice. Histologically, the MMP19-deficient 

mice showed reduced infiltration of neutrophils into the colonic 

tissue during the course of colitis but increased lesion 

extent, ulceration and oedema. The differences between WT 

and knockout mice were even more striking in the recovery 

phase of the colitis as MMP19-deficient animals showed only 

65 % survival. During the chronic colitis just 50 % survived and 

they did not regain their initial weight. MMP19-deficient animals 

showed elevated levels of pro-inflammatory cytokines in blood 

plasma and supernatants of ex vivo cultures of colonic tissues 

during all phases of colitis. 

Conclusion: The exacerbation of the colitis in the MMP19deficient 

mice together with elevated concentrations of proinflammatory 

cytokines derived from colonic tissue suggests 

that MMP19 plays an essential role in maintenance of intestinal 

homeostasis. This may have important implications for the two 

major inflammatory conditions of the intestine in humans, 

Crohn’s disease and ulcerative colitis. 

Quadriplex model enhances urinebased 

detection of prostate cancer 

Jamaspishvili T. 1 , Kral M. 2 , Khomeriki I. 2 , 

Vyhnankova V. 2 , Mgebrishvili G. 1 , Student V. 2 , 

Kolar Z1 . and Bouchal J. 1 

1 Laboratory of Molecular Pathology and Department 

of Clinical and Molecular Pathology, Institute of Molecular 

and Translational Medicine, Faculty of Medicine and Dentistry, 

Palacky University and University Hospital, Olomouc, 


2 Department of Urology, University Hospital, Olomouc, 


Background: The major advantages of multiplex urinebased 

assays are their noninvasive character, ability to monitor 

prostate cancer (CaP) with heterogeneous foci and detect 

cancer more accurately than do single marker tests. They might 

supplement serum PSA test which has low specificity resulting 

in a number of unnecessary biopsies especially in „grey zone“ 

and thereby reducing complications associated with biopsy. 

While the test for the prostate cancer gene 3 (PCA3) is commercially 

available, the aim of our research was to test other 

putative urine markers in multiplex settings (AMACR, EZH2, 

GOLM1, MSMB, SPINK1 and TRPM8). 

Materials and methods: Expression of the candidate biomarkers 

was studied in sedimented urine using quantitative 

RT-PCR on two sets of patients with PSA levels of 3–15 ng/ml 

(N=104) and 0.1–587 ng/ml (N=176). Total RNA isolation was 

performed by the Urine Exfoliated Cell RNA Purification Kit 

(Fisher Scientific, USA), quantified by Nanodrop, pretreated with 

Dnase I (Invitrogen) and reverse transcribed with SuperScript® 

III Reverse Transcriptase (Invitrogen), preamplified and tested on 

real-time PCR with Light Cycler® 480 (Roche). Clinicopathological 

characteristics were defined according to the WHO classifica- 


25

26 

ABSTRACT BOOK 

tion. Statistical analysis was done in SPSS using Spearman, 

Mann-Whitney and ROC analysis. Univariate and multivariate 

regression analysis were used to determine predicted probability 

of the individual markers and their combinations. 

Results and discussion: We confirmed that PCA3 is an independent 

predictor of cancer in the patients without restriction 

of serum PSA values (0.1–587 ng/ml). However, AMACR was the 

only parameter that differentiated CaP from nonCaP patients 

in patients with serum PSA between 3 and 15 ng/ml. The area 

under curve (AUC) for this gene was 0.645 with both sensitivity 

and specificity at 65 %. Further improvement was achieved by 

multivariate logistic regression analysis which identified novel 

duplex (TRPM8 and MSMB), triplex (plus AMACR) and quadriplex 

(plus PCA3) models for the detection of early prostate 

cancers (AUC 0.665, 0.726 and 0.741, respectively). 

Conclusions: In the current study we demonstrated that 

novel qudriplex test could be implemented as an adjunct to 

serum PSA or urine PCA3 and this could improve decision making 

for diagnostics in the case of “PSA dilemma” patients. 

This work was supported by grants NS 9940–4 from the 

Czech Ministry of Health and MSM 6198959216 from the 

Czech Ministry of Education and EU infrastructure support 

CZ.1.05/2.1.00/01.0030. Tamar Jamaspishvili was also supported 

by GACR 303/09/H048 from the Grant Agency of the Czech 

Republic and LF_2010_006. 

Comparison of proliferating activity 

in terminal villi of normal and diabetic 

placenta 

Jirkovská M. 1 , Kučera T. 1 , Jadrníček M. 1 , Niedobová V. 1 , 

Moravcová M. 2 , Krejčí V. 2 , Žižka Z. 2 

1 Institute of Histology and Embryology, First Faculty 

of Medicine, Charles University in Prague, Prague, 


2 Department of Obstetrics and Gynecology, First Faculty 

of Medicine, Charles University in Prague, Prague, 


Introduction: Placenta is a unique organ growing during 

its whole existence. In the last trimester it is expressed first 

of all by development and growth of terminal placental villi 

and their capillary bed. Comparing the normal and diabetic 

term placenta we have found out a higher degree of capillary 

branching in diabetic terminal villi. 

Aim: In this pilot study we tested the hypothesis that this 

phenomenon is connected with higher proliferating activity 

of cells constituting capillary wall. 

Material and methods: Specimens collected by systematic 

random sampling from five normal placentas and five placentas 

form pregnancies complicated by type I diabetes were fixed 

in formalin and embedded in paraffin. Histological sections 

were cut from five randomly chosen blocks per placenta and 

immunohistochemical detection of Ki-67 antigen as a marker 


of proliferation was performed. The number of Ki-67 positive 

nuclei was counted in trophoblast, stroma and capillary wall 

and the area of terminal villi was measured in 20 fields of view 

per section using Leica IM 500 program. 

Results: The Ki-67 labeled nuclei occurred in cytotrophoblast, 

villous stromal cells and in cells of capillary wall. The mean 

numbers of positive nuclei in cytotrophoblast, stromal cells as 

well as in cells of capillary wall per squared millimeter of section 

were found higher in diabetic placentas but the results were 

not significant due to great variability of data. 

Conclusions: Larger groups of placentas have to be assessed 

in order to compare the proliferating activity in norma and 

diabetes, and to test the correlation of this parameter with 

the level of metabolic control, mode of delivery and sex of 

newborn. 

Supported by the GAČR, project number 304/09/0733. 

A novel transgenic mouse model to 

study epidermal differentiation and 

wounding 

Kasparek P., Suchanova S., Krenek P., Sedlacek R. 

Department of Transgenic Models of Diseases, Institute 

of Molecular Genetics AS CR, Prague, Czech Republic 

Introduction: A number of skin diseases and pathological 

conditions such as dermatitis, psoriasis, or skin wounding are 

characterized by abnormal maturation and proliferation of 

keratinocytes. Thus, monitoring of keratinocyte differentiation 

in vivo appears to be a valuable tool to study processes of 

epidermal physiology and pathology. To address this issue, we 

generated a transgenic mouse model expressing a fluorescent 

marker tdTomato under the control of modified human involucrin 

promoter. Involucrin is well established differentiation 

marker being expressed in suprabasal layers of epidermis and 

in hyperproliferative keratinocytes of healing wounds. 

Aim: To create a model to study patho-physiological processes 

in vitro and in vivo in the outmost layers of epidermis. 

Materials and methods: DNA construct containing truncated 

promoter and first intron of human involucrin, cDNA 

of tdTomato, and polyA signal was subcloned and used for 

generation of transgenic mice by pronuclear microinjection 

on C57Bl/6N background. Skin from mouse ears and paws was 

cryosectioned and the expression of the transgene compared 

to the expression of other differentiation and proliferation markers 

using involucrin, keratin 14, keratin 6 antibodies. Model of 

irritant dermatitis was induced by application of croton oil. Skin 

wound was induced on mouse ear by linear 3 mm-puncher. To 

study regulation of involucrin promoter in vivo, transgenic mice 

were depilated and analyzed by whole-body imaging. 

Results: We have generated a transgenic mouse expressing 

tdTomato, a fluorescent reporter, under the control of human 

involucrin promoter. To assess the specific targeting and regulation 

of transgene, multiple organs were isolated from positive

animals and analyzed for tdTomato signals using fluorescent 

microscopy. The truncated human involucrin promoter with 

its intron targeted the transgene efficiently and specifically 

into the uppermost epidermal layers as well as into epithelia of 

the tongue and bladder. Detail analyses of skin sections from 

transgenic animals revealed specific expression of tdTomato 

in differentiated keratinocytes. Induction of irritant dermatitis 

on mouse ear led to an increase of fluorescent signal in 24 

hours. In the model of wound healing the fluorescent reporter 

was clearly upregulated 24 hours post wounding and this 

augmented expression lasted for additional 96 hours with the 

peak of expression around 72 hours. These observations are in 

accordance with previously reported changes of epidermal differentiation 

during acute dermatitis and healing of skin wounds. 

Fluorescence signal of the reporter can be easily followed in 

vivo using whole-body imaging. 

Conclusions: The transgenic mouse expressing tdTomato reporter 

under the human involucrin promoter could be used as 

a valuable tool to study processes leading to the dysregulation 

of epidermal barrier. Keratinocytes undergoing rapid proliferation 

and differentiation are in transgenic animals characterized 

by increase of fluorescent signal, which can be easily detected 

in vivo. This novel mouse model can be further used to follow 

an effect of various therapeutic agents for treatment of skin 

diseases in vivo. 

Asporin modulates tissue 

microenvironment, invasive growth 

and bone metastasis of breast cancer 

Kharaishvili G. 1 , Cizkova M. 2 , Bouchalova K. 2 , 

Sedlakova E. 1 , Kolar Z. 1 , Bouchal J. 1 

1 Laboratory of Molecular Pathology of Institute of Pathology, 

Institute of Molecular and Translational Medicine, Faculty 

of Medicine and Dentistry, Palacky University, Olomouc, 


2 Laboratory of Experimental Medicine, Pediatric Clinics 

of Medical Faculty of Palacky University and Faculty Hospital, 


Introduction: In an earlier publication we identified asporin 

as a novel cancer related protein (BMC Cancer 2007; 7: 55), 

also reported among bone metastasis specific genes (Cancer 

Letters 2009; 276: 212–220). Asporin is a member of the class 

I small leucine-rich repeat proteoglycan family and besides 

affinity to collagen, it binds calcium and initiates collagen 

mineralization. Asporin is abundantly expressed in tissues 

with high remodeling rate (e.g. periodontal ligament) whereas 

extremely high turnover of matrix proteins is also associated 

with invasive cancer. 

Material and methods: Formalin-fixed paraffin-embedded 

tissues [invasive ductal (101) and lobular (39) carcinomas 

classified according to hormone receptors and Her2; 5 bone 

metastases from autopsies] were immunohistochemically 

ABSTRACT BOOK 

stained with in-house polyclonal peptide antibody against 

asporin. The antibody was validated on uterus tissue as a positive 

control together with a commercial antibody (Abcam). 

Data were evaluated by non-parametric Mann-Whitney U test 

and Spearman correlation coefficient. Expression of asporin in 

both tissues and cell lines was also queried in databases Gene 

Expression Omnibus, ArrayExpress and Oncomine. Expression 

of asporin was further tested by RT-PCR in selected breast 

cancer cell lines. Migration and invasion of MDA-MB-231 cells 

were tested using real-time cell analysis Xcelligence system 

(Roche). Transwells were coated by solidifying collagen for 6 

hours (with or without recombinant asporin). Starved cells were 

seeded on the collagen in transwells while FBS was used as 

a chemoattractant in bottom wells. 

Results: Levels of asporin were significantly higher in invasive 

lobular carcinomas than ductal ones (p

28 

ABSTRACT BOOK 

pression profiling as breast cancer related protein. It has been 

reported to affect Wnt signaling, collagen deposition and bone 

formation and generally it belongs to extracellular matrix proteins 

which are abnormally expressed in tumor microenvironment. 

As CTHRC1 has not been studied by immunohistochemistry 

in breast cancer yet we decided to verify its expression in our 

patients set together with other relevant proteins. 

Material and methods: Formalin-fixed paraffin-embedded 

tissues of 173 invasive carcinomas (21 with metastases in bone, 

29 with other distant metastases), 27 lymph node metastases, 

36 benign and 31 normal cases were stained for proteins 

CTHRC1, periostin, versican, E-cadherin and beta-catenin using 

immunohistochemistry. Invasive carcinomas were classified 

according to hormone receptors and Her2 expression 

into luminal, triple negative and Her2 molecular subtypes. 

Specimens were assessed semiquantitatively by histoscore. 

Statistical analysis was performed with SPSS software using 

Spearman correlation, Wilcoxon and Mann-Whitney U test and 

Cox proportional hazard function. 

Results: Unless otherwise specified below, CTHRC1 was 

predominantly localized in cancer cell cytoplasm and extracellular 

matrix, while versican and periostin were expressed mostly 

by peripherial areas of infiltrating carcinoma and stromal cells. 

Expressions of CTHRC1, versican and periostin were significantly 

higher in breast cancer tissue in comparison to normal 

(p

cells in vessels per mm2 of villous cross-sectional surface, while 

in those with poor compensation this value was 18,8±22,18/ 

mm 2 . Also this difference was not statistically significant. 

Discussion: We found that number of apoptotic cells in 

placental vessels from DM type I pregnancies as well as in 

normal pregnancies is rather variable. For this reason a significant 

difference between these groups was not determined. 

Regarding vascular changes previously observed in DM type 

I placentas it could be concluded that these arise as a consequence 

of neovessel formation, but without significantly 

increased apoptosis of cells within the vessel wall. 

Double immunohistochemical staining 

in routine diagnostic pathology 

Lodererova A., Honsova E., Voska L., Gabris V., 

Maluskova J. 

Department of Pathology, Institute for Clinical and 

Experimental Medicine, Prague, Czech Republic 

Combining two different scientific disciplines – morphology 

and immunochemistry – immunohistochemistry has developed 

as an important instrument in research and clinical pathology. 

However, in some cases there is a need for knowledge 

about the relative localization of targets, which can only be 

obtained by vizualizing all relevant targets in one slide. Multiple 

staining can be defined as the detection of two or more targets 

on one slide. This technique enables the depiction of two or 

more targets/antigens in connection with other morphological 

features. We use the Benchmark ULTRA system in our lab, which 

is the next generation in IHC/ISH staining instrumentation. For 

the double staining method we use sequential staining. The 

most important step is to optimize the pretreatment step (time, 

buffer or protease) for both antigens. For the first detection 

step we selected ultraView Universal DAB system (brown) and 

for the second one ultraView Universal Alkaline Phosphatase 

Detection Kit (red). 

The following combinations of antigens were determined 

as helpful: Ki 67 + CK 7; Ki 67 + CD 31; CD 34 + CK 7; CD 31 + p 

53; D2- 40 + AE 1/3; CD 3 + CK 7; κ + λ light chains. 

Conclusion: Detection of more targets/antigens in one 

slide has good results. Moreover, this method spares the tissue, 

which is very important in cases with small biopsy samples. 

ABSTRACT BOOK 

MicroRNA Assessment as a New 

Diagnostic and Prognostic Tool of 

Barrett´s Esophagus. Pilot Study 

Luzna P. 1 , Gregar J. 2 , Uberall I. 3 , Prochazka V. 2 , 

Ehrmann J. jr. 1,3 

1 Department of Histology and Embryology, Faculty of Medicine 

and Dentistry, Palacky University and University Hospital 

Olomouc, Czech Republic (pavla83@hotmail.com) 

2 nd II Internal Clinic, Faculty of Medicine and Dentistry, 

Palacky University and University Hospital Olomouc, 


3 Laboratory of Molecular Pathology, Faculty of Medicine 

and Dentistry, Palacky University and University Hospital 


Backround: Barrett´s esophagus (BE) is the disease which 

can progress through several stages into the adenocarcinoma 

of the esophagus (EAC). Since 1993 the process of carcinogenesis 

has been investigated in connection with regulators of 

gene translation – microRNAs. It was found that among them 

miR-21, miR-192, miR-196a and miR-203 were often related to 

progression and prognosis of various tumors. Therefore, the 

aim of this pilot study was to assess the role of these miRNA 

in BE in various stages of progression. 

Design: 30 patients diagnosed with BE (15 cases without 

dysplasia, 10 cases with low grade dysplasia and 5 cases with 

high grade dysplasia/adenocarcinoma) were examined. 5 cases 

of normal esophagus were used as a control. Samples from 

paraffin blocks were cut into the slides, specific lesions were 

microdissected, total RNA was isolated and microRNA assays 

were carried out. Their levels were relatively quantified to two 

endogenous controls RNU44 and RNU48. 

Results: We found significant increase of miR-192 levels in 

all stages of BE compared to the control. The levels of miR-203 

were downregulated in all cases and moreover negatively 

correlated with the progression of disease. miR-21 expression 

was increased in all cases without dysplasia and in all cases 

with low grade dysplasia, however in cases with high grade 

dysplasia/adenocarcinoma it was not changed. Conversely, 

miR-196a was amplified in all adenocarcinoma cases and not 

in dysplastic lesions. 

Conclusion: Our results suggest that detection of miR-192 

may be important for primary diagnosis of Barrett´s esophagus. 

Moreover, we found out that detection of miR-203, miR-21 

and miR-196a may play role in assessment of progression of 

BE. Therefore, it seems that miRNA assays may serve as a new 

diagnostic and prognostic tool for BE. 

Acknowledgement: This work was supported by Grant of 

Ministry of Health of Czech Republic, No. NS 10279–3 and by 

Grant MSM 6198959216. 


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ABSTRACT BOOK 

Prognostic factors in multiple myeloma 

Mareckova J.¹,Scudla V.², Ehrmann J.¹, Abrahamova P.¹ 

¹ Department of Pathology, Laboratory of Molecular Pathology, 

Faculty of Medicine and Dentistry, Palacky University 

Olomouc, Czech Rebublic 

2 3 rd Internal clinic, Faculty of Medicine and Dentistry, 

Palacky University Olomouc, University Hospital Olomouc, 


Introduction: Multiple myeloma is plasma cells neoplasm 

characterized by serum monoclonal immunoglobulin called 

the M component and skeletal destruction with osteolytic 

lesions, pathologic fractures, bone pain, hypercalcemia and 

anaemia. It is usually incurable with median of survival of 3 

years and 10 % at 10 %. Multiple myeloma (MM) is connected 

to a number of chromosomal abnormalities, in most cases 

IgH translocation with FGFR3, CCND1, CCND2, CCND3, c-MAF 

in early stages. Further chromosomal changes appear with 

the progression of the disease the most frequent of which 

are monoallelic deletion or monosomy of chromosome 13, 

trisomies of chromosome 8, 9, 15 and many others. It has also 

been revealed that some proteins controlling cell the cycle 

and apoptosis (p53, p16, FGFR3, cyclin D 1, 2, 3, Bcl-2, caspase 9) 

seem to play an important role during MM pathogenesis and 

progression. However, there are no reliable data on their prognostic 

significance in the various stages of disease (monoclonal 

gammapathy of uncertain significance – MGUS, smouldering 

MM and advanced MM). Therefore the aim of this pilot study 

was analysis of expression of these proteins in various stages 

of MM and potentially extend the panel of prognostic markers 

which allows differentiation of the above-mentioned disease 

stages. 

Material and methods: Bone marrow from 35 patients 

treated by the same chemotherapy protocol (VAD) and autologous 

transplantation were used. Standard indirect immunohistochemistry 

on formalin fixed, paraffin-embedded sections 

was used for the detection of p53, p16, FGFR3, cyclin D 1, 2, 3, 

Bcl-2, caspase 9 using a high temperature epitope retrieval 

technique. Immunohistochemical staining was evaluated by 

a semi-quantitative method using a histoscore which is the 

multiplication of positivity by intensity of staining. Intensity 

of staining was scored as weak (1), moderate (2) or strong 

(3) while positivity of staining was assessed as percentage of 

tumour cells. 

Results: Bone marrow samples of patients in advanced 

stages of MM showed high expression of Bcl-2 in tumor cells, 

in contrast to those in remission which showed weak or no 

positivity for Bcl-2. p53 and p16 were mostly completely negative. 

Only few patients with advanced disease showed strong 

positivity for p53. Caspase 9 was negative in biopsies taken 

before treatment but we detected several caspase 9 positive 

cells in patients after treatment. 

Conclusions: Based on decreased Bcl-2 expression in patients 

in remission, these preliminary data suggest that detection 

of low Bcl-2 expression in bioptic samples of MM might 


be used as positive prognostic factor. Negativity of p16 can 

be explained by hypermethylation which leads to p16 deactivation. 

Mutation of p53 is probably infrequent in multiple 

myeloma but the mechanism of its impaired function needs 

further study. 

This study was supported by IGA NR/9500–3 and UP 

LF_2011_009. 

Determination of metallothioneins and 

alpha-methylacyl-CoA racemase in 

tumor prostate diseases 

Masarik M. 1 , Gumulec J. 1 , Sztalmachova M. 1,2 , 

Hlavna M. 1 , Kuchtickova S. 1 , Rovny A. 3 , Hrabec R. 3 , 

Eckschlager T. 4 , Krizkova S. 2 , Adam V. 2 , Kizek R. 2 

1 Department of Pathological Physiology, Faculty of Medicine 

Masaryk University, Brno, Czech Republic 

2 Department of Chemistry and Biochemistry, Faculty of 

Agronomy, Mendel University, Brno, Czech Republic 

3 Department of Urology, St. Anne´s University Hospital, Brno, 


4 Department of Paediatric Haematology and Oncology, 

2nd Faculty of Medicine Charles University in Prague, 


Introduction: Metallothioneins (MT) belong to the group 

of intracellular, low-molecular cysteine-rich proteins with molecular 

weight from 6 to10 kDa. Due to their high affinity to 

heavy metal ions (Zn, Cd, As, etc.), their main and crucial function 

is homeostasis maintenance and detoxification of heavy 

metals. The role of MT in tumour tissue remains still unclear, 

but MT can be considered as new promising tumour marker. 

Alpha-methylacyl-CoA racemase (AMACR) is a -oxidation of 

branchedβperoxisomal and mitochondrial enzyme involved 

in the fatty acids and was recently reported that AMACR has 

very high sensitivity and specificity to prostate adenocarcinoma. 

Therefore is very important to find new approaches in 

AMACR detection. 

Aim: The main aim of this paper is to study expression of 

MT and AMACR in tumor prostate cell lines and in patients 

serum on protein and mRNA level. Moreover, we compare 

the differences in MT expression between cells influnced with 

zinc ions. 

Material and methods: Biological samples. Prostatic cell lines 

derived from prostate adenocarcinoma (LNCaP-FGC, PC-3, and 

22RVL) and prostatic cell line derived from normal epithelium 

(PNT1A – human immortalized prostatic cell line) were used. 

Protein detection. SDS-PAGE electrophoresis and Westernblot 

analysis with subsequent immunodetection were used. 

Immunohistochemical detection: Immunohistochemical detection 

of AMACR was performed by R.T.U Vectastain universal 

ABC kit (Vector Laboratories, Burlingame, CA, USA). 

Results: We investigated influence of zinc ions on MT expression 

in prostatic cell lines derived from prostate carcinoma

and MT level in patients serum samples. For the MT determination 

we used SDS-PAGE electrophoresis and western blot 

analysis with subsequent immunodetection. Moreover we 

used modern electrochemical methods (Brdicka reaction) for 

detection of MT. We demonstrated up-regulation of prostatic 

specific antigen expression in prostate tumour cells (LNCaP, PC- 

3, 22RVL) and contrariwise down-regulation of MT expression. 

This fact can be explain by decreased concentration of zinc 

ions in prostatic tumor cells and therefor lower concentration 

of MT in tumor cell lines in comparing with non-tumor cells. 

Besides immunodetection we measured MT by adsorptive 

transfer stripping technique coupled with differential pulsed 

voltammetry Brdicka reaction and we obtained similar results 

as in immunodetection. In case of AMACR, results show relatively 

big diferences between tumor and non-tumor cell lines 

in AMACR content in mRNA and protein level as well. In case 

of tumor cells the expression of AMACR was up-regulated in 

comparison with non tumour cells. Then we cultivated cells 

on glass slides for immunodetection of AMACR in situ and we 

obtained similar results. 

Conclusions: This study provides important new information 

about expresion of MT and AMACR in prostate tumor cell 

lines. Evidently MT expression is significantly down regulated 

in 22RVL, and PC-3 cells while AMACR was up regulated. We 

observed significant (α=0.05) difference in MT content between 

cell lines treated/nontreated with zinc ions by using 

immunohistochemical methods and electrochemical methods 

as well. 

Acknowledgements: This work was supported by the grants 

GACR 301/09/P436 and IGA MZ NS10200–3/2009. 

Identifi cation of grafted bone marrow 

cells in the recipient tissues 

Mokrý J. 1 , Filip S. 2 , Vávrová J. 3 , Čížková D. 1 , 

Šinkorová Z. 3 , Mičuda S. 4 

1 Department of Histology and Embryology, Charles University 

in Prague, Faculty of Medicine, Hradec Králové, Czech Republic 

2 Department of Oncology and Radiotherapy, Charles University 

in Prague, Faculty of Medicine and Teaching Hospital, Hradec 

Kralové, Czech Republic 

3 Department of Radiobiology, University of Defence Brno, 

Faculty of Military Health Sciences, Hradec Králové, 


4 Department of Pharmacology, Charles University in Prague, 

Faculty of Medicine, Hradec Králové, Czech Republic 

Introduction: Identification of transplanted cells is facilitated 

when donor cells were tagged with endogenous vectors. 

Aim: Comparison of efficacy of two different models based 

on bone marrow donor cells derived from transgenic ROSA26 

and eGFP mice. 

Material and methods: Full bone marrow or lin-CD117+ 

of B6;129S-Gt (ROSA) 26Sor mice or C57BL/6-Tg (CAG-EGFP) 

ABSTRACT BOOK 

C14Y01FM131Osb mice were transplanted into B6129SF2/J or 

C57BL/6J recipients subjected to 9Gy whole body lethal irradiation. 

At different time points post transplantation (30, 60 and 

180 days), cell settlement in recipient tissues including spleen, 

thymus, small intestine and liver was assessed histologically 

and verified by qPCR analysis. 

Results: By day 30 transplanted cells infiltrated stromal components 

of examined organs as connective tissue wandering 

cells. In the thymus and spleen, grafted bone marrow-derived 

cells participated in lymphatic infiltration of both organs including 

thymic cortex and splenic nodules, indicating generation 

of donor-derived T- and B-lymphocytes. Transplanted animals 

were allowed to survive up to 6 months, which demonstrated 

establishment of a stable cellular chimerism. Data from qPCR 

were in a correlation with histological analysis. 

Conclusion: eGFP mice represent a more reliable system 

over ROSA26 mice since GFP+ cells may be directly visualized 

in tissues (do not suffer from drawbacks associated with 

a poor penetration of X-Gal substrate and non-specifity of 

histochemical staining) and may be also directly analysed for 

flow cytometry. Histological examination of tissues obtained 

from transplanted mice is prior to PCR analysis because it allows 

not only to confirm identity, phenotype, status of transplanted 

cells and engraftment efficacy but also to specify their number, 

fate and distribution in recipient tissues. 

This work was supported by MSM 0021620820. 

Role of apoptosis associated genes 

in predicting clinical outcome 

of breast cancer 

Skálová H. 1 , Dundr P. 1 , Povýšil C. 1 , Velenská Z. 1 , 

Berková A. 1 , Staněk L. 1 , Dlouhá Z. 2 , Petruželka L. 2 , 

Tvrdík D. 1 

1 st Institute of Pathology, 1 Faculty of Medicine, Charles 

University and General University Hospital in Prague, 


2 st Department of Oncology, 1 Faculty of Medicine, 

Charles University and General University Hospital in Prague, 


Introduction: Neoadjuvant chemotherapy, also called preoperative 

chemotherapy, is now widely used in the treatment 

of breast carcinoma. It has several potential advantages 

compared with the traditional strategy of surgery followed 

by adjuvant chemotherapy. Neoadjuvant chemotherapy substantially 

reduces the size of the primary tumor and lymph 

node metastasis in grater then 80 % of cases, increasing the 

probability that breast-conserving surgery can be performed 

instead of mastectomy. Neoadjuvant chemotherapy is also 

a valuable research tool for discovery predictive markers of 

chemotherapy response. 


31

32 


ABSTRACT BOOK 

Aim: The study is aimed at investigation of the biomarkers 

in predicting pathologic response to the therapy. We assume 

that regression of the tumors after neoadjuvant treatment is 

executed predominantly by apoptosis despite different chemotherapy 

agents. 

Material and methods: The role of apoptosis in regression of 

the tumors after neoadjuvant chemotherapy was determined 

by two independent method: TUNEL and anti-active caspase 3 

assay, respectively. The transcriptional profile of 84 key apoptosis 

genes was evaluated in both pre-therapeutically obtained 

tumor tissue by core needle biopsy and in speciemen removed 

by final surgery, using Real-Time PCR assay (SABiosciences). 

Results: This study included 16 patients with histologically 

proven invasive breast cancer, that are primarily indicated to 

breast conserving surgery or mastectomy but that are treated 

by neoadjuvant chemotherapy. In each case, analysis of transcription 

profile was performed before and after the treatment, 

respectively. On the basis of hierarchical cluster analysis of 13 

significantly changed genes, we divided patients into good and 

bad prognosis groups which good correlate with progression 

free survival. In the good prognosis group we found statistically 

significant downregulation of expression of MCL1 and IGF1R 

genes after neoadjuvant treatment. We also found statistically 

significant overexpression of BCL2L10, BCL2AF1, CASP8, CASP10, 

CASP14, CIDEB, FADD, HRK, TNFRSF25, TNFSF8 and TNFSF7 

genes. In contrast, we found upregulation of IGF1R due to the 

treatment in the group of poor prognosis. The expression of 

remaining genes remained almost unchanged in this group 

of patients. 

Conclusion: Unfortunately, little progress has been made 

with regards to new molecular prognostic/predictive markers 

that can assist oncologists in treatment decision-making for 

breast cancer. In this study, we verify our presumption that 

tumor regression after chemotherapy is caused by apoptosis 

despite the diverse schema of the treatment. Moreover, we 

develop the 13 apoptosis associated genes expression assay 

which may be heplful for stratification of the patients into 

groups with different response to chemotherapy. As we shown, 

gene expression profiling after neoadjuvant chemotherapy is 

a valuable research tool for investigation of molecular markers 

which may better reflect tumor biology and treatment 

response than standard prognostic and predictive factors. 

This work was supported by Grant IGA MZ ČR No. NS10575–3. 

Decalcifi cation options of bone 

material for consequential molecular 

biological and cytogenetic diagnosis – 

from the experimental model to 

trepanobiopsy 

Staněk L. 1,2,3 , Tvrdík D. 1 , Střítský J. 1 , Lísová S. 1 , 

Berková A. 1 , Ludvíková M. 1,3 

1 Institute of Pathology, General Faculty Hospital and 

1st Faculty of Medicine, Charles University in Prague, Prague, 


2 Department of Experimental virology, IHBT, Prague, 


3 Institute of Biology, Faculty of Medicine in Pilsen, 

Charles University, Pilsen Czech Republic 

Molecular-biological and cytogenetic analysis of bone 

material has today a unique place in many areas of medicine. It 

is essential for the diagnosis of some lymphoproliferative disorders, 

and also for biomedical research. In biomedical research, 

the analysis concerns mainly the tissue engineering field, where 

mesenchymal stem cells differentiated into osteoblasts are 

used as a substitute in bioinplantology. Diagnostic in pathology 

includes especially the diagnosis of lymfoproliferative 

disease, mainly lymphomas, including both histological and 

immunohistochemical examination. But subtyping on the basis 

of cytogenetic and molecular-biological methods, detection 

of faults, translocations or amplification, is necessary. 

For bioptical analysis and diagnostic ultra thin slices are 

required. Quality of these slices is dependent on superior material 

decalcification. However, most decalcifying techniques 

(e.g. formic acid) irreversibly damages the DNA and blocks 

subsequent molecular subtyping. Formic acid protons purine 

ring and thereby reduces the glycosidic bond to form covalent 

bonds. Furthermore, in the tissue degrades DNA to a string of 

less than 650 bp. 

For decalcification of trepanobioptic rollers – diagnosis 

and experimentally for a mouse femur (Mus musculus) EDTA/ 

NaOH (10 mol/l – 40 %) protocol was used (EDTA – ethylenediaminetetraacetic 

acid CH2N (CH2CO2H) 2,2 – polyaminokarboxylic 

acid, which binds divalent cations). NaOH reacts with 

acid in form of ethylendiamine acid disodium salt (EDTA). This 

salt is more soluble in water than the acid itself. In reaction 

with metal ions (Mn+) such as Ca2+ complex ions of (MY) type 

are formed. We did the decalcification for three days on the 

shaker, changing EDTA every five hours and then rinse with 

water. Subsequently, the tissue was processed according to 

the protocol and paraffin blocks were prepared. 

The trepanobioptic rollers were processed by FISH using 

different Visis FISH probes (Abbott), to determine the translocations. 

DNA purification using QIAamp DNA Mini Kit (Qiagen) 

and subsequent DNA amplification using the IdentiClone IGH 

Gene Clonality Assay kit (InVivoScribe) for the detection of the 

Ig heavy chains genes remodeling was performed.

A mouse femur was cut up and classical histological H/E 

staining was done to verify bone structure preservation. 

Afterwards Visis probe LSI BCR/ABL Dual Color Translocation 

EC Probe (Abbott) was used for cytogenetic examination. 

All molecular methods were fully applicable and highly 

reproducible with very good visualization on specimens decalcified 

with this method. It can be concluded that this decalcification 

method is gentle, with tissue well preserved for routine 

histological staining and DNA remains in a condition suitable 

for further molecular biological and cytogenetic analysis. 

Role of PPARγ in colon cancer cells 

Straková N. 1,2,3 , Hofmanová J. 1,3 , Tylichová Z. 1,3 , 

Knopfová L. 4 , Šmarda J. 4 , Kozubík A. 1,3 , Ehrmann J. 2,3 , 

Kolář Z. 2,3 

1 Laboratory of Cytokinetics, Institute of Biophysics, Acad Sci 

Czech Rep, v.v.i., Brno, Czech Republic 

2 Laboratory of Molecular Pathology, Faculty of Medicine and 

Dentistry, Palacky University, Olomouc, Czech Republic 

3 Biomedical Centre of Institute of Biophysics Acad Sci Czech 

Rep, v.v.i. and Faculty of Medicine and Dentistry, Palacky 

University, Olomouc, Czech Republic 

4 Laboratory of Cell Differentiation, Institute of Experimental 

Biology, Masaryk University, Brno, Czech Republic 

Colorectal cancer (CRC) is the third most commonly diagnosed 

cancer in Western countries. The treatment of CRC 

is focused on identifying gene networks involved in regulation 

of cell growth, differentiation and cell death. PPARs 

(Peroxisome Proliferator – Activated Receptors) are nuclear 

hormone receptors. After binding of ligand to receptor, PPAR 

forms a heterodimeric complex with retinoid X receptor which 

then binds to PPAR response element on DNA. This interaction 

is responsible for the regulation of genes involved in glucose 

and lipid metabolism, differentiation and apoptosis. There are 

three PPAR isoforms: PPARα, PPAR β/δ and PPARγ. The expression 

of PPARγ has been described in both normal and many 

different cancer cell types. In colon tumors, increased PPARγ 

level was detected. Thus, our interest was focused on the role 

of PPARγ in regulation of colon tumorigenesis. 

First, we compared the expression of PPARγ in 11 human 

colon cancer cell lines with different grade of malignancy by 

Western blotting. The highest expression was observed in 

adenocarcinoma HT-29 cells. Expression of PPARγ was confirmed 

by immunocytochemistry fluorescent staining. Next, we 

analyzed the effects of PPARγ synthetic ligands – thiazolidinediones 

(rosiglitazone, ciglitazone) and compared them with 

the effects of short chain fatty acid – sodium butyrate (NaBt). 

Butyrate is produced by gastrointestinal tract by anaerobic 

fermentation of fiber and can modulate proliferation, differentiation, 

and apoptosis of colon cancer cells. Rosiglitazone 

and ciglitazone decreased proliferation of colon cancer cell 

lines in dose-dependent manner. Interestingly, real-time cell 

analysis using xCELLigence demonstrated that HCT-116 p21- 

ABSTRACT BOOK 

/- were more sensitive to rosiglitazone/ciglitazone treatment 

than wild type HCT-116. NaBt treatment dose-dependently 

reduced proliferation of colon cancer cell lines, too. And finally, 

the Luciferase assay was performed to analyze PPARγ activation 

and it detected that rosiglitazone, ciglitazone as well as 

NaBt activated this receptor. Moreover, NaBt increased PPARγ 

activity much more intensively than rosiglitazone or ciglitazone 

in colon cancer cell lines studied. 

In conclusion, colon cancer cell lines studied expressed 

PPARγ with different intensity. Rosiglitazone, ciglitazone as well 

as NaBt decreased cell proliferation and activated PPARγ. NaBt 

appeared as more potent activator of PPARγ than rosiglitazone/ 

ciglitazone in colon cancer cell lines. 

This work was supported by grant IGA MZ ČR NT/11201–5 

and grant Czech Science Foundation No. 7301/11/1730. 

Molecular genetics changes in 

melanocystic lesions – case report 

Uvírová M. 1 , Dvořáčková J. 2 , Šimová J. 1 , Urbanovská I. 1 , 

Žiak D. 1 , Konvalinka D. 1 , Kubová B. 1 

1CGB laboratory, Ostrava, Czech Republic 

2Faculty of Medicine University of Ostrava, Ostrava, 


Introduction: The worldwide incidence of malignant melanoma 

is increasing annually more than 4 %. Some forms of 

nevocellular lesions are a diagnostic problem in morphological 

terms; particularly in cases of pigmented lesions of benign 

nature, such as Spitz naevus. A correct determination of biological 

nature of the neuroectodermal tumour is one of the basic 

prerequisites for a proper treatment. Histological examination 

is the gold standard to establish the diagnosis. Although many 

cases may be reliably classified according to current histopathological 

criteria, there is a group of cases in which an agreement 

has not been reached even among experts. Molecular genetic 

studies show that the difference between melanomas and 

benign nevocellular lesions consists of a multiplication or loss 

of specific chromosomal regions. In melanomas, using the CGH 

method, the most frequent deletions were found in regions 

6q, 8p, 9p and 10q, and the most frequent amplifications in 

regions 1q, 6p, 8q, 17q, 20q and over the entire chromosome 

7. Based on these studies a combined probe Vysis LSI RREB1/ 

LSI MYB/LSI CCND1/CEP 6 (Abbott, USA) was developed, which 

can be used to detect numeric alterations associated with 

malignant melanoma as a diagnostic aid for formalin fixed, 

paraffin embedded sections. 

Aim: Case report 

Material and methods: Nevoid skin lesion of the left arm of 

seventy years old woman, 10 mm in diameter, was removed 

in 2006. Material was sent for histological examination with 

a diagnosis of benign naevus. Conclusion of the histological 

examination was dysplastic familiar naevus. In april 2010, reexcision 

of the scar recurrence was performed. Clinical diagnosis 


33

34 

ABSTRACT BOOK 

defined as malignant melanoma. Conclusion of histological 

examination was supperficial spreading melanoma. 

For the identification of genetic changes associated with 

diagnosis of malignant melanoma in the sample from the year 

2010 Multi Colour FISH (Fluorescence In Situ Hybridisation), the 

combined probe Vysis LSI RREB1/LSI MYB/LSI CCND1/CEP 6 

(Abbott, USA) has been applied. The same assay was used in 

the sample from the year 2006. 

Results and conclusions: Using Interphase FISH method in 

the sample from the year 2010 (histology: supperficial spread 

melanoma) pathological finding was proven – amplification 

of gene CCND1 (lokus 11q13) in all examined tumour cells. We 

examined 30 nonoverlapping cells from 3 different areas of the 

tumour. In the material from the year 2006 we found isolated 

cells with the same aberration as in the first examined sample 

from the year 2010 – the amplification of gene CCND1. 

While, using the current histopathological criteria, the sample 

was diagnosed in 2006 as a rather dysplastic familiar naevus, 

genetic changes associated with the diagnosis of malignant 

melanoma in isolated cells were also found. Molecular testing 

seems to be a useful tool for diagnosing skin lesions. 

Role of matrix metalloproteinase-19 

in liver fi brosis 

Zbodakova O. 1 , Jirouskova M. 1 , Sedlacek R. 1 , Ehrmann J. 2 , 

Hajduch M. 3 , Jirkovska M. 4 

1 Laboratory of Transgenic Models of Diseases, Institute of 

Molecular Genetics of the ASCR, Prague, Czech Republic 

2 Department of Pathology, Medical Faculty, Palacky University, 


3 Laboratory of Experimental Medicine, Department of 

Pediatrics and Oncology, Palacky University and University 

Hospital, Olomouc, Czech Republic 

4 Institute of Histology and Embryology, First Faculty of 

Medicine, Charles University, Prague, Czech Republic 

Introduction: Liver fibrosis is a complex process in which the 

interplay between diverse factors affects the outcome of the 

disease. Several matrix metalloproteinases (MMPs) have been 

shown to play a role in fibrosis development and its resolution. 

Their functions include degradation of basal membrane, release 

of growth factors and degradation of collagen deposits during 

the resolution of fibrosis. MMP-19 appears to be constitutively 

and highly expressed in the liver. However, nearly nothing is 

known about its function in this organ. 


Aim: To investigate role of MMP19 in the development and 

resolution of liver fibrosis. 

Material and methods: Using MMP19-deficient mice, we 

studied involvement of MMP19 in the development and 

resolution of liver fibrosis in an experimental model of CCL4 

administration. 8 weeks old WT and MMP19-deficient males 

were injected with CCl4 for 6 weeks. Samples were collected 

at peak of fibrosis (48h after last injection) or 10 and 15 days 

later to follow the resolution of fibrosis. Serum samples were 

analyzed to monitor liver function (ALT, AST, ALP, billirubin) and 

chemokine levels (MCP1, KC, IL6). Samples of left lateral lobe 

were collected and further processed using αTNF staining 

with H&E, Sirius red, and several antibodies. Concurrently, liver 

samples were also snap-frozen for further analyses including 

mRNA analysis, HPLC analysis of hydroxyproline content, and 

immunoblotting. 

Results: Both MMP19-deficient and wildtype (WT) mice 

developed pronounced liver fibrosis with deposition of fibrillar 

collagens after 6 weeks of chronic intoxication. Activity of aminotransferases 

in serum was highly elevated in all challenged 

animals compared to physiological conditions. However, the 

ALT and AST levels were significantly lower in MMP19-deficient 

mice compared to WT animals. Furthermore, the concentration 

of chemokines KC and MCP-1in sera of MMP19-deficient 

mice was lower than in control mice. Liver of both groups 

SMA staining, indicating activation of hepatic-exhibited strong 

positivity for stellate cells that are the main collagen-producing 

cell type in the liver. However, the localization was distinct 

in MMP19-deficient mice (mainly pericentral localization) in 

comparison to WT controls that exhibited staining along the 

central-portal connections. RNA chip analysis revealed 3.5 

times higher expression of MMP13 in MMP19-deficient mice 

compared to WT animals. This enzyme is typically expressed 

in first stages of fibrotic development with second peak later 

during the recovery process. 

In the recovery phase, serum levels of hepatocyte damage 

markers and chemokines were already in normal values in both 

mouse strains, however, fibrosis was still high in both WT and 

MMP19-deficient mice after 10 days of recovery. Nevertheless, 

Sirius red staining revealed ongoing fibrosis in MMP19-deficient 

animals 5 days later while livers of wild-type mice showed healing 

process with minimum of collagen deposition. 

Conclusion: Altogether our results indicate that MMP19deficientmice 

display different kinetics in development and 

resolution of liver fibrosis in CCL4 induced model. Our finding 

suggests that MMP19 deficiency leads to slower progression 

in development of fibrosis as well as slower resolution of liver 

fibrosis during recovery stage.

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NOTES 

PROGRAMME AND ABSTRACT BOOK

HLAVNÍ SPONZOR 

SPONZOŘI 

edesa

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