Volume 51 (2005) No. 4

Volume 51 (2005) No. 4
Dedication
Milestones in the Field of Tumour Microenvironment – Contributions and Perspectives of Professor Isaac P. Witz
O. LEVY-NISSENBAUM.........................................................................................85
The Department of Cell Research and Immunology, The George S. Wise Faculty of Life Sciences, Tel-Aviv University, Tel Aviv, Israel
Corresponding author: Orlev Levy-Nissenbaum, The Department of Cell Research and Immunology, The George S. Wise Faculty of Life Sciences, Tel-Aviv University, 69978, Tel Aviv, Israel. E-mail: orle@post.tau.ac.il.
No abstract available.
Full text. 85-88
Original Articles
Intranucleolar Translocation of AgNORs in Early Granulocytic Precursors in Chronic Myeloid Leukaemia and K 562 Cells
K. SMETANA, H. KLAMOVÁ, M. PLUSKALOVÁ, P. STÖCKBAUER, I. JIRÁSKOVÁ,
Z. HRKAL...................................................................................................................89
Institute of Haematology and Blood Transfusion, Prague, Czech Republic
Corresponding author: Karel Smetana, Institute of Haematology and Blood Transfusion, U Nemocnice 1, Prague 2, 128 20 Czech Republic. E-mail: karel.smetana@uhkt.cz
Abstract.
Full text. 89-92
Characteristics of Two Mouse bcr-abl-Transformed Cell Lines. II. Pathological Lesions Induced in Mice
F. JELÍNEK1, 2 , E. SOBOTKOVÁ1, V. VONKA1.......................................................93
1Department of Experimental Virology, Institute of Hematology and Blood Transfusion,
Prague, Czech Republic
2Veterinary Histopathological Laboratory, Prague, Czech Republic
Corresponding author: František Jelínek, Veterinary Histopathological Laboratory, Sojovicka 16, 197 00 Prague 9, Czech Republic.
E-mail: f.jelinek@email.cz.
Abstract.
Full text. 93-102
Viability of Human Chondrocytes in an ex Vivo Model in Relation to Temperature and Cartilage Depth
M. DROBNIČ1, T. MARŠ2, A. ALIBEGOVIĆ3, V. BOLE4, J. BALAŽIC3, Z. GRUBIČ2, J. BRECELJ1........................103
1Department of Orthopaedic Surgery, University Medical Centre, Ljubljana, Slovenia
2Institute of Pathophysiology and 3Institute of Forensic Medicine, Medical Faculty, University of Ljubljana, Slovenia
4Economic Institute at the Law School, Ljubljana, Slovenia
Corresponding author: Matej Drobnič, Department of Orthopaedic Surgery, University Medical Centre, Zaloška 9, SI – 1000 Ljubljana, Slovenia. Tel.: (+386) 1 522 4174; Fax: (+386) 1 522 2474; e-mail: matej.drobnic@mf.uni-lj.si
Abstract.
Full text. 103-108
Immunogenicity and Protective Effect of a DNA Construct Encoding Certain Neutralizing Epitopes of Herpes Simplex
Virus Type-1 Glycoprotein B
T. BAMDAD1, M. H. ROOSTAEE1, M. SADEGHIZADEH2, F. MAHBOUDI3, A. KAZEMNEJAD4,
H. SOLEIMANJAHI1.......................................................................................109
1Department of Virology, School of Medical Sciences, 2Department of Genetics, School of Basic Sciences, Tarbiat Modarres University, Tehran, Iran
3Department of Biotechnology, Pasteur Institute, Tehran, Iran
4Department of Biostatistics, School of Medical Sciences, Tarbiat Modarres University, Tehran, Iran
Corresponding author: Taravat Bamdad, Department of Virology, School of Medical Sciences, Tarbiat Modarres University,   P.O.Box  14115-111, Tehran, Iran. Tel.: +98 21 8011001 ext 3880;   Fax: +98 21 8013030; Bamdad_T@modares.ac.ir
Abstract.
Full text. 109-113
Review
Molecular Therapeutics - Lessons from the Role of Src in Cellular Signalling
Z. TUHÁČKOVÁ................................................................................................114
Department of Intracellular Communications, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic
Corresponding author: Zdena Tuháčková, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Flemingovo nám. 2,
166 37  Praha 6, Czech Republic. Tel.: (420) 220 183 287; Fax: (420) 233 320 702; e-mail: tuhack@img.cas.cz
Abstract.
Full text. 114-120
Original Articles
Intranucleolar Translocation of AgNORs in Early Granulocytic Precursors in Chronic Myeloid Leukaemia and K 562 Cells
K. SMETANA, H. KLAMOVÁ, M. PLUSKALOVÁ, P. STÖCKBAUER, I. JIRÁSKOVÁ,
Z. HRKAL

The present study was undertaken to provide missing information on the distribution of AgNORs in large nucleoli of human leukaemic early granulocytic precursors in vivo as well as in vitro.  In vivo, the distribution of AgNORs was studied in early granulocytic precursors of
patients suffering from chronic myeloid leukaemia who were both untreated and treated with imatinib mesylate. AgNORs were visualized by silver reaction under conditions which facilitated to see their distribution by light microscopy. In vitro, the distribution  of AgNORs was studied
in proliferating and ageing  K 562 cells which originated from chronic myeloid leukaemia. In vitro, the ageing of K 562 cells produced intranucleolar translocation of AgNORs to the nucleolar periphery. Such translocation was also observed in some leukaemic early granulocytic precursors in vivo, e.g. in bone marrow myeloblasts and promyelocytes of leukaemic patients. As was expected, the intranucleolar translocation of AgNORs in early granulocytic precursors was more frequent in patients treated with the cytostatic therapy – imanitib mesylate. The above- mentioned findings suggest that myeloblasts and promyelocytes with AgNORs translocated to the periphery of large nucleoli might be in the ageing state, similarly as blastic cells of leukaemic myeloid origin (K 562 cells)  in ageing cultures. Thus, the translocation of AgNORs might be
a useful marker of premature ageing in the future and might contribute  to the  evaluation of the single cell state under various experimental as
well as clinical conditions. However, more clinically oriented studies are required in this direction.  
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Characteristics of Two Mouse bcr-abl-Transformed Cell Lines. II. Pathological Lesions Induced in Mice
F. JELÍNEK, E. SOBOTKOVÁ, V. VONKA

Groups of six BALB/c mice each were intravenously inoculated with lethal doses of Ba-P210 (B210) or 12B1 cells and  examined by  autopsy, histology, special staining methods, enzyme histochemistry and immunohistochemistry. Clinical symptoms related to neoplasia consisted of a poor nutritional state, anaemia, mild to moderate dehydration and apathy. Paresis was apparent in three mice inoculated with 12B1 cells. Necropsy revealed splenomegaly in all animals. Sporadic haemorrhages in the lungs and enlargement of some lymph nodes were seen in some of the animals. Histological examination showed neoplastic cells in the spleen, in the bone marrow of the sternum, in the lung interstitium and in sinusoids of the liver in all mice. In six of nine brains examined, mild to moderate infiltration by neoplastic cells was observed. In all but two mice mild infiltration of the kidneys was found. The enlargement of lymph nodes was caused by an accumulation of neoplastic cells. The paresis was due to neoplastic infiltration of the vertebra, epidural space and  spinal roots.  Staining with Sudan black revealed cytoplasmic granules in neoplastic cells; however, the peroxidase reaction was negative. Numerous neoplastic cells disseminated in the red pulp of the spleen were reactive with CD3, CD79β, CD11b and with  neutrophil antibodies. We  classified the disease induced by both of the cell lines as acute myeloid undifferentiated leukaemia (AML MO).
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Viability of Human Chondrocytes in an ex Vivo Model in Relation to Temperature and Cartilage Depth
M. DROBNIČ, T. MARŠ, A. ALIBEGOVIĆ, V. BOLE, J. BALAŽIC, Z. GRUBIČ, J. BRECELJ

Chondrocytes in human articular cartilage remain viable post-mortem. It has however not been established yet how the storage temperature affects their survival, which is essential information when post-mortem cartilage is used for toxicologic studies. Our aim was to construct a simple model of explanted knee cartilage and to test the influences of time and temperature on the viability of chondrocytes in the ex vivo conditions. Osteochondral cylinders were procured from the cadaveric femoral condyles. The cylinders were embedded in water-tight rubber tubes, which formed separate chondral and osteal compartments. Tubes were filled with normal saline, without additives, to keep chondrocytes under close-to-normal conditions. The samples were divided into two groups stored at 4°C and 35°C, respectively. Three samples of each of these two groups were analysed at the time of removal, and then three and nine days later. Images of Live-Dead staining were scanned by a confocal laser microscope. Count of viable chondrocytes in four regions, from surface to bone, was obtained using image analysis software. The regression model revealed that the number of viable chondrocytes decreased every day by 19% and that an increase in temperature by 1°C decreased their viability by 5.8%. The temperature effect fell by 0.2 percentage points for every 100 μm from the surface to the bone. Herein we demonstrate that chondrocytes remain viable in the ex vivo model of human knee cartilage long enough to be able to serve as a model for toxicologic studies. Their viability is, however, significantly influenced by time and temperature.
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Immunogenicity and Protective Effect of a DNA Construct Encoding Certain Neutralizing Epitopes of Herpes Simplex
Virus Type-1 Glycoprotein B
T. BAMDAD, M. H. ROOSTAEE, M. SADEGHIZADEH, F. MAHBOUDI, A. KAZEMNEJAD,
H. SOLEIMANJAHI

Much attention is presently focused on the vaccination with certain epitopes of an antigen. To further study the ability of neutralizing epitopes mapped in the first 1515 nucleotides of glycoprotein B of herpes simplex virus type-1 (gB-1) to induce neutralizing antibodies, a DNA immunization approach was employed.  Vaccination of mice with a plasmid expressing the neutralizing epitopes induced humoral immune responses, although the antibody titre was significantly lower than that of antibodies induced by the full-length gB-1 gene. Furthermore, the plasmid DNA could not protect the mice against HSV-1 lethal challenge, but could significantly prolong the survival time compared to mock- vaccinated group.
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Review
Molecular Therapeutics - Lessons from the Role of Src in Cellular Signalling
Z. TUHÁČKOVÁ

Src is the oldest and best studied protooncogene participating in normal cells in proliferation, maintenance of normal intercellular contacts and cell motility. Its discovery opened the way for fundamental discoveries on the cell cycle, cell growth and death, cell-cell signalling, cell morphology and motility, and cancer biology. Src is poorly transforming, consistent with a role as a normal cellular protooncogene that, when activated, might serve as an oncogene. Its activation promotes growth during the process of tumorigenesis by stimulating the proliferation of pre-cancerous cells and also regulates other activities such as adhesion and invasion during the later stages of tumour progression. Overexpression of the Src protein and the increase of its specific protein kinase activity have been observed in various human malignancies and linked to the development of cancer and tumour progression to distant metastases. These observations have led to the recent rediscovery of Src, a molecule that has been investigated during nearly a century and that shows promise as a new target for therapy of human cancer in the development of anticancer therapeutics. To determine the role of Src in human cancer, which is still not fully understood, molecular details of many pathways that intersect with Src are necessary to be uncovered. A detailed map of signalling pathways regulating signal transduction and signal integration induced by Src may identify location of different „checkpoints“ for the therapeutic intervention of human diseases due to the altered activity of Src.
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