Volume 47 (2001) No. 3

Volume 47 (2001) No. 3

Articles
Patterns of Deletions and the Distribution of Breakpoints in the Dystrophin Gene in Czech Patients with Duchenne and Becker Muscular Dystrophy (Statistical Comparison with Results from Several Other Countries)
I. HRDLIČKA1, J. ZADINA2, R. KREJČÍ3, A. SRBOVÁ1, M. KUČEROVÁ1...............................81
1Department of Medical Genetics, Thomayer University Hospital, Prague, Czech Republic
2Institute of Rheumatology, Prague, Czech Republic
3Department of Genetics and Microbiology, Faculty of Science, Charles Universty, Prague, Czech Republic
Corresponding author: Ivan Hrdlička, Department of Medical Genetics, Thomayer University Hospital, Vídeňská 800, Prague 4-Krč, Czech Republic. Tel: +420 (2) 61083619; Fax: +420 (2) 4728227; e-mail: hrdlicka@ftn.cz.
Abstract.
Full text. 81-82 83-84 85-87


Supernumerary Human Preembryos Provide Potential for Preimplantation Genetic Diagnosis
S. M. DELIMITREVA, R. S. ZHIVKOVA, I. T. VATEV.................................88
IVF-ET Centre "Technobioassistance", Department of Biology, Medical Faculty, Sofia, Bulgaria
Corresponding author: Stefka M. Delimitreva, IVF-ET Centre "Technobioassistance", Department of Biology, Medical Faculty, 1, "St. G. Sofiyski" Str., 1431 Sofia, Bulgaria. Fax: +359 (2) 51 86 31; e-mail: delimitr@medfac.acad.bg.
Abstract.
Full text. 88-89 90-91


Effects of Combined Treatment of Rats with Cadmium and Ionozing Radiation on Nucleic Acids in the Kidneys, Liver and Haemopoietic Organs
L. SLOVINSKÁ, K. KROPÁČOVÁ, M. KOLESÁROVÁ, E. MIŠÚROVÁ........................92
Department of Cellular and Molecular Biology, Faculty of Sciences, P. J. Šafárik University, Košice, Slovakia
Abstract.
Full text. 92-93 94 95 96 97-98 99-100


High Incidence of Chromosome Aberrations after Radiochemotherapy for Hodgkin's Disease: a Report of a Case and a Review of the Literature
H. LALIC1, B. RADOSEVIC-STASIC2, C. VOLAVSEK3...................................101
1Department of Occupational and Environmental Medicine, 2Department of Physiology and Immunology, Medical Faculty, University of Rijeka, Croatia
3Department of Medical Cytogenetic and Toxicology, Centre for Environmental Health, Ljubljana, Slovenia
Corresponding author: Hrvoje Lalic, 51000 Rijeka, Brentinijeva 5, Croatia. Tel.: +385 51 371 094; 098 328 110; e-mail: biserr@mamed.medri.hr.
Abstract.
Full text. 101-102 103 104-105


Monoclonal Antibody register
Monoclonal Antibodies Specific to Polynuclear Aromatic Hydrocarbons
M. SUCHANEK1, T. SCHARNWEBER2, M. FISHER3, D. KNOPP2, R. NIESSNER2...................106
1EXBIO Praha a.s., Prague, Czech Republic
2Institute of Hydrochemistry, Technical University Munich, Germany
3Technion-Israel Institution of technology, Haifa, Israel
Corresponding author: Miloslav Suchanek, EXBIO Praha a.s., Videnska 1083, 14220 Praha 4, Czech Republic. Fax: +420 (2) 475 2151; e-mail: suchanek@exbio.cz.
Background.
Full text.106-107


Monoclonal Antibodies Specific for Bovine CD18
R. DUSINSKY, J. TOMASKOVA, L. HOROVSKA, M. SIMON...............................................108
Institute of Animal Biochemistry and Genetics, Slovak Academy of Sciences, Ivanka pri Dunaji, Slovakia 
Corresponding author: Roman Dusinsky, Institute of Animal Biochemistry and Genetics, Slovak Academy of Sciences, Moyzesova 61, 900 28 Ivanka pri Dunaji, Slovakia. Tel.: +421 (07) 4594 3882; fax: +421 (07) 4594 3932; e-mail:ubgz-dusi@savba.sk.
Background.
Full text. 108 109-110


Articles
Patterns of Deletions and the Distribution of Breakpoints in the Dystrophin Gene in Czech Patients with Duchenne and Becker Muscular Dystrophy (Statistical Comparison with Results from Several Other Countries)
I. HRDLIČKA, J. ZADINA, R. KREJČÍ, A. SRBOVÁ, M. KUČEROVÁL. 

Deletion pattern analysis of the dystrophin gene was performed in 115 unrelated Czech patients with Duchenne and Becker muscular dystrophy. In 50 patients (43.5% of  the analysed patients) exon deletions were detected by routinely performed multiplex PCR for 18 selected exons and for the area of muscle-specific promoter of the dystrophin gene. All startpoints and endpoints of deletions (100 breakpoints) were detected using PCRs for another 29 exon areas of the dystrophin gene (altogether primers for 47 different exons were used). Most of the breakpoints were found in introns 44 (16% of breakpoints), 47 (14%) and 50 (8%). The comparison of distributions of breakpoints in the area of the main hot spot of the dystrophin gene (introns 43–52) was made (chikvadrat test in a contingency table) in six different populations from the Czech Republic, Bulgaria, Hungary, Italy, Turkey and India. In compared populations, statistically significant differences were found by the pooled test. No significant difference  between the  Czech population and other studied populations was found by pair comparisons. On the other hand, pair comparisons revealed significant differences between populations from Bulgaria and Hungary, Bulgaria and Turkey, Hungary and Italy. The results of the presented study support  the theory suggested by other authors that specific differences in intron sequences of the dystrophin gene can exist between different populations, possibly as a result of a genetic drift. 
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Supernumerary Human Preembryos Provide Potential for Preimplantation Genetic Diagnosis
S. M. DELIMITREVA, R. S. ZHIVKOVA, I. T. VATEV

Preimplantation genetic diagnosis is an alternative to the classical prenatal diagnosis for couples undergoing in vitro fertilization. It allows very early embryo selection – before the intrauterine embryo transfer. Prior to clinical application of preimplantation diagnosis in the Infertility Treatment Centre “Technobioassistance”, Sofia Medical Faculty, we have developed preimplantation diagnosis models of human spermatozoa and untransferred 2-8-cell human embryos obtained in vitro. Directly fluorescein isothiocyanate-labelled probes specific for the centromeric regions of chromosomes 1, 5, 19 and X (Boehringer Mannheim) were used. Eighty-six point three percent of  fixed blastomeres with normal size and shape had  unfragmented nuclei with dispersed interphase chromatin or mitotic chromosomes and all of them demonstrated successful hybridization. In cases with more than 75% of embryo cells suitable for analysis we were able to estimate the presence of mosaicism among the blastomeres.
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Effects of Combined Treatment of Rats with Cadmium and Ionozing Radiation on Nucleic Acids in the Kidneys, Liver and Haemopoietic Organs
L. SLOVINSKÁ, K. KROPÁČOVÁ, M. KOLESÁROVÁ, E. MIŠÚROVÁ

The influence of Cd (1 mg/rat CdCl2 i.p.) and/or gamma radiation (6 Gy) on RNA and DNA content and/or concentration in the intact kidney and hypertrophic kidney (on the 44th hour after UN) and in other slowly and quickly proliferating organs was studied. The period between administration of Cd and Ir in the group with combined treatment was 30 min, between treatment (administration of Cd, Ir and combination of  both treatments - Cd + Ir) and UN it was 1, 7, 14 and 21 days.
The total extent of damage caused by the treatments in the investigated organs was following: intact kidney < liver < hypertrophic kidney and bone marrow < spleen < thymus. In the intact and hypertrophic kidney and liver, the administration of Cd caused more extensive changes in comparison with gamma irradiation and the effects of combination of the treatments were similar to those of Cd alone. In the bone marrow, spleen and thymus, more profound changes were observed after Ir in comparison with Cd administration, and the effects of  combined treatment were similar to the effects of Ir alone. The changes in the hypertrophic kidney after administration of Cd and/or Ir were more extensive than in the intact kidney, which suggests latent injury induction in the rat kidney by these noxa. The higher effectiveness of the treatments in the hypertrophic kidney than in the intact one was manifested mostly by the decrease in the RNA and DNA content, which was mainly due to inhibition of growth induced by UN and not by a real decrease in DNA and RNA contents caused by loss of damaged cells.
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High Incidence of Chromosome Aberrations after Radiochemotherapy for Hodgkin's Disease: a Report of a Case and a Review of the Literature
H. LALIC, B. RADOSEVIC-STASIC, C. VOLAVSEK

To emphasize the importance of information obtained by conventional cytogenetic tests, in this study we will present a case report showing the development of HD in a young engineer of medical radiology, who had been professionally exposed to ionizing radiation during 4 years. Bio-dosimeter data and chromosomal aberration analysis made in a group of colleagues working in the same ionizing conditions excluded the possibility that she was overexposed to ionizing radiation, but retroactive analysis showed  that at the time of employment she had a moderately increased level of chromosomal aberrations in peripheral blood lymphocytes (4.6%), with higher than normal incidence of chromatid breaks (22 x 10-3 per cell) and dicentric fragments (5 x 10-3 per cell). After the treatment of lymphoma by chemotherapy in combination with radiotherapy she also demonstrated a very high post-therapeutic level of  chromosomal aberrations (21.5%), which consisted mostly of increased dicentric fragments (65 x 10-3 per cell). Although her illness is now clinically cured, the observed genotoxic changes point to a greater risk for delayed complications after HD, emphasizing the necessity for  further continuous survey of this patient. 
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Monoclonal Antibody Register
Monoclonal Antibodies Specific to Polynuclear Aromatic Hydrocarbons
M. SUCHANEK, T. SCHARNWEBER, M. FISHER, D. KNOPP, R. NIESSNER

The interest in polycyclic aromatic hydrocarbons (PAHs), especially benzo[a]pyrene, in environmental samples (e.g. city aerosols) is derived from their known mutagenic potential (Topinka et al., 1998) and their adjuvant effect for the causation of allergies (Diaz-Sanchez, 1997; Bömmel et al., 2000). Analysis of PAHs is routinely performed by gas chromatography/mass spectrometry (GC/MS) or high performance liquid chromatography/fluorescence detection (HPLC/FD). However, these methods are time consuming, mainly due to complex procedures involved in the preparation of samples, which usually includes removal of disturbing matrix compounds and concentration of the target analytes. The objective of our project is to develop a rapid immunoassay which can be applied to screening of a large number of samples without the need for demanding sample preparation. In this communication we describe monoclonal antibodies BAP?13 and BAP?14 specific for a range of PAHs. 
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Monoclonal Antibodies Specific for Bovine CD18
R. DUSINSKY, J. TOMASKOVA, L. HOROVSKA, M. SIMON

The molecule CD18 is a subunit of leukocyte adhesion molecules belonging to the beta2 integrins (CD11/CD18). Recently, free forms of intracytoplasmic CD18 molecules have been described (Drbal et al., 2000). Integrins are polypeptide heterodimers consisting of a common 95-kD beta chain (CD18) non-covalently linked to a unique alfa subunit: 180-kD alfaL (CD11a), 170-kD alfaM (CD11b), or 150-kD alfaX (CD11c). The molecule CD11a/CD18 (LFA-1) is primarily expressed on lymphocytes, CD11b/CD18 (MAC-1) on neutrophils and monocytes, CD11c/CD18 on monocytes and macrophages (Gahmberg et al., 1997). The integrins are type I membrane proteins containing an N-glycosidic carbohydrate (Asada et al., 1991). The extracellular domains of integrins mediate cell-matrix and cell-cell contacts, while their cytoplasmic tails associate with the cytoskeleton. Integrins are not active in resting cells, but need activation to become adhesive. The CD18 molecule has been shown to play a central role in regulating activity (Green et al., 1998). The beta2 integrins bind to intercellular adhesion molecules ICAM and to several soluble proteins, many of which are involved in inflammation (Gahmberg et al., 1998). Their pivotal importance is best evident in individuals lacking functional CD11/CD18 due to mutation in the CD18 gene with LAD (leukocyte adhesion deficiency) syndrome in man (Arnaut, 1990) or BLAD (bovine leukocyte adhesion deficiency) syndrome in cattle (Kehrli et al. 1990; Shuster et al., 1992). This syndrome is characterized by repeated infections.
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