Vol. 3 No. 4 December 1997

Volume 3 (1998) pp 3-11
Title INCREASED ACTIVITY OF PHOSPHOLIPASE C IN AORTAS OF SPONTANEOUSLY HYPERTENSIVE RATS CORRELATES WITH DECREASED SPHINGOMYELIN : TOTAL PHOSPHOLIPID RATIO
Authors E. Gryckiewicz, A. Dettlaff and T. Pawełczyk*
Abstract The activity of phospholipase C (PLC) in the aortas of spontaneously hypertensive rats (SHR) was higher than in the aortas of age-matched normotensive Wistar-Kyoto rats (WKY). This was associated with the higher level of inositol 1,4,5-trisphosphate and diacylglycerol in aortas of SHR compared to the level of these compounds found in aortas of WKY. Observed changes in PLC activity correlates with changes in phospholipid composition of SHR aortas. The sphingomyelin (SM) to total phospholipid ratio decreased significantly in aortas of SHR compared to WKY. Since, SM was proposed to be the major PLC d inhibitor in vivoit might be possible that observed higher activity of PLC in aortas of SHR results from decreased content of this phospholipid.
Address and Contact Information Department of Clinical Biochemistry, Medical University of Gdańsk, 80-211 Gdańsk, Poland
*To whom correspondence should be adressed. Tel. (058) 347-82-22 ext. 17-76, Fax (058) 344-96-53, e-mail; tkpaw@amedec.amg.gda.pl
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Volume 3 (1998) pp 13-20
Title ARSENICAL - INDUCED TRANSCRIPTIONAL ACTIVATION OF THE YEAST Saccharomyces cerevisiae ACR2 AND ACR3 GENES REQUIRES THE PRESENCE OF THE ACR1 GENE PRODUCT
Authors P. Bobrowicz and S. Ułaszewski*1
Abstract The yeast ACR1 gene encodes a putative transcriptional regulatory bZIP protein involved in the arsenical resistance. We demonstrate that the ACR2 and ACR3 genes are positively regulated by Acr1p. Changes in the ACR1 gene dosage influence the ACR2-lacZ and ACR3-lacZ fusion genes response to the arsenite induction. Arsenic and antimony compounds, but not bismuth and cadmium salts, strongly induced the expression of the ACR3 gene encoding the arsenite transporter related to the prokaryotic ArsB proteins.
Address and Contact Information Institute of Microbiology, Wroclaw University, Przybyszewskiego 63, 51-148 Wroclaw, Poland
* To whom correspondence should be addressed. TEL: 48-71-3247293; FAX: 48-71-3252151; E-mail: stan@microb.uni.wroc.pl
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Volume 3 (1998) pp 21-24
Title EASY PCR SCREENING OF Pichia pastoris TRANSFORMANTS
Authors D. Arora, A. Chauhan and N. Khanna*
Abstract A simple protocol for easy PCR screening of P. pastoris transformants is described. In short, the P. pastoris cells are lysed with very small amount of the enzyme Zymolase and the crude cell lysate is directly used in PCR. This protocol needs no tube transfer steps and also obliviates the requirement of freezing the samples at -800C before PCR screening. Because of a single step screenig, both overall and actual hands on time are considerably reduced.
Address and Contact Information 1 International Centre for Genetic Engineering and Biotechnology, P.O. Box 10504, Aruna Asaf Ali Marg, New Delhi - 110 067, India
*To whom correspondence should be addressed
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Volume 3 (1998) pp 25-35
Title IMMUNOSPECIFIC PROTEIN WITH MOL. WT OF 38/39 kDa FROM LYMPHOCYTIC LEUKEMIC CELLS
Authors Z. M. Kiliańska*1, A. Ptasińska1, J. Błoński2, J. Chruściel1, P. Szymczyk1, E. Krykowski2 and T. Robak2.
Abstract One dimensional patterns of proteins from homogenates and four cellular fractions i.e., nuclear, mitochondrial, microsomal, and cytosolic from normal and leukemic lymphocytes were compared. Results obtained revealed that the neoplastic transformation of normal lymphocytes into CLL and ALL ones is associated with the expression of some novel proteins. Electrophoretically-specific nuclear protein of B-CLL lymphocytes with mol. wt of 38/39 kDa was used as immunogen to produce rabbit antiserum. It was observed that obtained antiserum crossreacted with 38/39 kDa antigen of nuclear fractions from CLL and ALL lymphocytes (15 of 16 studied), but not with any of normal ones. It was shown, by Western blot technique, that the expression of 38/39 kDa antigen is correlated with progression of B-CLL disease.
Address and Contact Information 1Department of Cytobiochemistry, University of Łódź, Banacha 12/16, 90-237 Łódź, Poland,
2Department of Haematology, Medical University of Łódź, Paderewskiego 4, 93-513 Łódź, Poland.
*Reprint address
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Volume 3 (1998) pp 37-48
Title PURIFICATION OF THE Ca2+ - ATPase FROM RAT CORTICAL, CEREBELLAR AND HIPPOCAMPAL SYNAPTOSOMAL MEMBRANES BY TWO - STEP CHROMATOGRAPHY.
Authors L. Żylińska*, L. Lachowicz and E. Gromadzińska
Abstract In Southern blot analysis of Paramecium genomic DNA using ß-adrenergic specific oligonucleotides labelled with Digoxigenin-ddUTP, the DNA species of the same molecular size were detected by different molecular probes. The 6.5 kb DNA species hybridized with the probes directed to the adjacent ß2-receptor transmembrane domains: to TM3 - including Asp 113 involved in ß-agonists and antagonists binding, and to TM4 - the oligonucleotide being one of the "gene-specific universal mammalian sequence-tagged site". Since a profound physiological effect of beta-agonists and antagonists has been previously observed in Paramecium, such a hybridization pattern may suggest an existence of Paramecium DNA sequences homologous to those of metazoan eukaryotes encoding for a membrane beta-adrenergic receptor.
Address and Contact Information Nencki Institute of Experimental Biology, Pasteura 3, 02-093 Warsaw, Poland
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Volume 3 (1998) pp 389-397
Title DNA BREAKING ACTIVITY OF THE PESTICIDE MALATHION AND ITS METABOLITE ASSAYED BY THE SINGLE CELL GEL ELCTROPHORESIS (COMET ASSAY)
Authors J. Blasiak1, A. Trzeciak1, P. Jaloszynski2, K. Szyfter2, R. Osiecka3, A. Blaszczyk3
Abstract Genotoxicity of pesticides is of a special significance because of the common presence of these agents in the environment and the long latent period between the exposure and effects becoming apparent. The DNA damaging effects of the commonly used organophosphorus insecticide malathion and its major metabolite malaoxon were evaluated. Freshly isolated human peripheral blood lymphocytes were incubated with 75 and 300 mM of malathion or malaoxon for 1 h. Sensitive alkaline microgel single cell electrophoresis (comet assay) was used to assess damage to the lymphocyte DNA. Possible lesions detected by this technique could be single- and double-strand DNA breaks as well as alkali-labile sites. Malaoxon, unlike malathion, induced DNA damage in a dose-dependent manner. The effect of malaoxon at 300 µM was comparable with the effect of hydrogen peroxide at 20 µM. Well recognized genotoxic effects of malathion may be a consequence either of the interaction of its major metabolite malaoxon with DNA or use of technical grade of malathion which contains impurities, including malaoxon. However, humans are exposed to the pesticide of technical grade and have the ability to metabolize malathion to malaoxon and in this regard malathion used as an organophosphorus insecticide can be considered as a genotoxic substance.
Address and Contact Information 1 Department of Molecular Genetics and 3 Department of Plant Cytology and Cytochemistry, University of Lodz, Poland
2 Institute of Human Genetics, Polish Academy of Sciences, Poznań, Poland
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Volume 3 (1998) pp 399-409
Title POTENTIAL SUPPRESSOR GENE LOCI AND HUMAN PROSTATE NEOPLASMS
Authors M. Brys and W. M. Krajewska
Abstract The biology of prostate cancer is still poorly understood. Allelic loss studies indicate that there likely exist multiple sites harbouring candidate tumour suppressor genes (TSG), some of which may have an important role in primary tumours, and some in late stages of prostate cancer. The recent studies on the localization of potential TSG in neoplastic transformation of prostate comprise chromosome regions 7q, 8p, l0p/q, 16q, 17q, and 18q. In connection with accumulation of genetic changes affecting functioning of critical TSG, the multistep cancer progression hypothesis is a useful starting point in efforts to understand the biology of the neoplastic lesions of prostate.
Address and Contact Information Department of Cytobiochemistry, University of Łódź, Poland
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Volume 3 (1998) pp 411-419
Title LONG-CHAIN ORCINOL HOMOLOGS FROM CEREAL BRAN ARE EFFECTIVE INHIBITORS OF GLYCEROPHOSPHATE DEHYDROGENASE.
Authors J. Rejman and A. Kozubek*
Abstract It was found that 5-n-alk(en)ylresorcinols isolated from high-fiber diet, wheat or rye bran, effectively inhibit glycerol-3-phosphate dehydrogenase (GPDH, EC 1.1.1.8), the key enzyme of triacylglycerol synthesis in adipocytes. The concentration of alk(en)ylresorcinols necessary for 50 per cent inhibition of GPDH vary from 3.8 µM to 6.4 µM. The effectiveness of the inhibition depends on the length of alk(en)enylresorcinol side chain and hydrophilic ring substitutions. The effectivity of homologs with long side chain (C17 and C19 ) in apparent inhibition of GPDH is slightly higher than that reported previously for adipostatin A (5-n-pentadecylresorcinol) isolated from Streptomyces (Tsuge, N., Mizokami, M., Imai, S., Shimazu, A., & Seto, H. (1992) J. Antibiot. 45, 886-891). The fact that the studied resorcinols are natural components of high-fiber human diet, such as porridge and bran products, suggests their possible participation in prevention of the risk of corpulence.
Address and Contact Information University of Wroclaw, Institute of Biochemistry and Molecular Biology Przybyszewskiego 63/77, 51-148 Wroclaw, Poland
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Volume 3 (1998) pp 421-429
Title STABLE GENETIC TRANSFORMATION OF MATURE WHEAT EMBRYOS USING SILICONE CARBIDE FIBERS AND DNA IMBIBITION
Authors W. Sawahel+@ and M. Saker*
Abstract Improvement of wheat (Triticum aestivum L) by biotechnological approaches is currently limited by a lack of tissue culture - free transformation system.  In this paper, we present the development of a gene transfer system using DNA imbibition [DI]; and/or silicone carbide fibers [SCF] treatments for gene delivery and seed-derived embryos as gene target.  Plasmid DNA (pBI221.23), containing the selectable "hpt" gene for hygromycin resistance and the reporter "gus" gene, was delivered into mature wheat embryos via [DI] and/or [SCF] treatments. In a histochemical analysis of ß-GUS activity, an average of 13.8 % of the mature embryos receiving the combined treatment [DI + SCF] showed gus expression, whereas only 3.2% and 0.4% showed gus expression following [SCF] and [DI] respectively. The mature embryos also showed more multi-site gus expression (4.6%) than those treated only with [SCF] (0.4%). The growth of the produced plantlets on a medium containing 45 µg/ml hygromycin B indicated that the transformation efficiency of the combined treatments [DI + SCF] was 0.8% in comparison to 0.1% for  that  of  [SCF] treatment  and no hygromycin   resistant-plantlets were  detected following [DI]. Stable transformation was confirmed through polymerase chain reaction "PCR" and Southern hybridization which indicated that a functional hpt gene had integrated into wheat chromosomal DNA.
Address and Contact Information Department of Microbial Genetics+ & Department of Plant Cell and Tissue Culture,*National Research Center, Cairo, Egypt
@-corresponding author
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Volume 3 (1998) pp 431-437
Title GENETIC ANALYSIS OF MAIZE ANTHER CULTURE RESPONSE
Authors J.C. Song, L.J. Jiang, Q.B. Wang, S.Y. Wang and F.F. Guo
Abstract Genetic barrier remains one of the major difficulties in maize anther culture and its application in breeding procedure. In order to investigate the genetic effects of anther culturability, five maize inbred lines with different anther culturability, including world-wide used commercial lines Mo17 and B73, were selected to form a 5 x 5 complete diallel cross. 1500-2000 anthers from each of the 25 combinations were cultured on modified Yupei medium. 0-20.93 embryoids per 100 plated anthers were produced from different combinations. Inbred line A3 had the highest anther culturability and produced at least 5 embryoids per 100 anthers in any involved combinations. Hayman's diallel analysis results showed that both embryoid and plant yields are genetically controlled. The additive effects were predominant while the dominant effects were significant only for plant yield. The dominance was partial and the dominant genes had the positive effects on anther culturability. The narrow sense heritability (h2N) are 0.97 and 0.96 for embryoid and plant yields respectively. Taken together, these results suggested that the genetic improvement of maize anther culturability might be efficient using hybridisation and/or selection procedures.
Address and Contact Information Department of Agronomy, Shandong Agricultural University Taian, 271018 China
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Volume 3 (1998) pp 439-448
Title NUCLEAR MATRIX LOCALIZATION OF CHICKEN HSP108.
Authors F. Altieri, M. Eufemi, B. Maras and C. Turano
Abstract A glycoprotein purified from the internal nuclear matrix of chicken liver nuclei has been identified, by partial sequencing and by Western blotting, as hsp108, which is a component of the hsp90 superfamily. This protein has been previously characterized as a protein which copurifies with the cytoplasmic progesterone receptor and as a transferrin-binding protein of the chicken oviduct cell membrane. We have found that hsp108 is present in the nuclear matrix even in the absence of a heat shock or of other noxious conditions. Some of the properties already described for its homologous proteins (endoplasmin, grp94, hsp100) might explain its function at the nuclear matrix level. Hsp108 isolated from the liver nuclear matrix has a carbohydrate composition significantly different from that of the protein of the oviduct cell membrane.
Address and Contact Information Department of Biochemical Sciences 'Alessandro Rossi Fanelli' and C.N.R. Center of Molecular Biology, University La Sapienza, Rome, Italy.
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Volume 3 (1998) pp 449-466
Title ON THE SELF-SIMILARITY OF THE LOGISTIC MAP
Authors Z.J. Grzywna1,* , L.S. Liebovitch2 and Z. Siwy1
Abstract The self-similarity of a quadratic, one parameter logistic map was shown. Depending on the value of a constant, four different patterns were obtained and analysed. Only for the chaotic region (R = 4) the obtained patterns were almost independent of the resolution used (the frequency of probing). Some of the analytical operations used as standard tools in investigating the maps properties were also analysed and suitably altered, where necessary.
Address and Contact Information 1 Department of Physical Chemistry and Technology of Polymer Section of Physics, Mathematics and Computer Science Silesian Technical University, 44-100 Gliwice, Strzody 9, Poland
2 Center for Complex Systems, Florida Atlantic University, Boca Raton, Florida 33431, USA

*  to whom all correspondence should be addressed.
1 E-mail: grzywna@zeus.polsl.gliwice.pl Phone: +48 32 37 17 52  Fax: +48 32 37 17 22,  371509
2 E-mail: liebovitch@walt.ccs.fau.edu  Phone: +01 407 367 2239  Fax: +01 407 367 2223
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