Vol.8 No.2 June 2003

Volume 8 (2003) pp 261-268
Title THE GENETIC CHARACTERISTICS SACCHAROMYCES CEREVISIAE ACI+ MUTANTS
Authors Renata Grochowalska1, Beata Machnicka1, Robert Wysocki2 And Tadeusz M. Lachowicz1
Abstract A series of 30 Saccharomyces cerevisiae aci+ mutants (characterized as acidifying Ogur's glucose medium containing bromocresol purple) were isolated after EMS mutagenesis. All the mutants excreted acid metabolites to the medium after 24 or 48 hours of incubation. The character of the aci+ mutations was defined using classical genetic techniques. Three of the aci+ mutants were studied by molecular genetics techniques.
Address and Contact Information 1Institute of Biotechnology and Environmental Protection, University of Zielona Góra, Monte Cassino 21b, 65-001 Zielona Góra, Poland,
2Institute of Microbiology, University of Wrocław, Przybyszewskiego 63/77, 51-148 Wrocław, Poland
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Volume 8 (2003) pp 269-278
Title SYNDECAN-4 DISTRIBUTION DURING THE DIFFERENTIATION OF SATELLITE CELLS ISOLATED FROM SOLEUS MUSCLE TREATED BY PHORBOL ESTER AND CALPHOSTIN C
Authors Edyta Brzóska, Iwona Grabowska, Edyta Wróbel and Jerzy Moraczewski
Abstract It was shown that syndecans have a potential role in muscle development. We focused this study on the role of syndecan-4 distribution and phosphorylation during the differentiation of satellite cells isolated from Soleus muscle. Syndecans are cell surface heparan sulfate proteoglycans (HSPGs) that bind numerous ligands through their HS glycosaminoglycan chains (GAG). They play a role in cell-extracellular matrix and cell-cell adhesion, signal transduction and the targeting of growth factors and other molecules to the cell surface. Syndecan-4 acts as a co-receptor or, along with integrins, is localized to the cell membrane of focal contacts. Syndecan-4 participates in the organization of the structure of focal contacts reacting with extracellular matrix molecules. The interaction of syndecan-4 with protein kinase C (PKC) isoforms is the main mechanism regulating its distribution in cells. Our current study focused on the role of the distribution of syndecan-4, and its interactions with PKC isoforms during the differentiation of activated satellite cells. We used the PKC activator TPA (12-O-tetradecanoyl phorbol 13-acetate) and the PKC inhibitor Calphostin C (Cal C). We concluded that syndecan-4 was important not only in the activation of satellite cells, but also in myoblast differentiation. During our research, we observed the presence of syndecan-4 and changes in its location over the course of that process. We also showed that TPA and Cal C treatment had an influence on the subcellular distribution of syndecan-4, but there was no influence on myoblast differentiation. We speculated that the reason for changes after TPA treatment was the interactions with activated PKCα, which provoked syndecan-4/PKCα complex translocation to integrins. We also supposed that Cal C treatment inhibited PKCd activity and probably induced PKCa association to syndecan-4, and syndecan-4 translocation to integrins.
Address and Contact Information Department of Cytology, Faculty of Biology, Warsaw University, Miecznikowa 1, 02-096 Warsaw, Poland
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Volume 8 (2003) pp 279-284
Title THE ACTIVITY OF LIPOXYGENASE IN Arabidopsis thaliana (L.) Heynh- A PRELIMINARY STUDY
Authors Ewa Skórzyńska-Polit and Zbigniew Krupa
Abstract The activity of lipoxygenase (EC 1.13.11.12) in Arabidopsis thaliana (L.) Heynh seedlings and mature plants was estimated spectrophotometrically at 234 nm. Linoleic acid was used as a substrate. Lipoxygenase activity showed two pH optima: at 7.0 and 10.0 in seedlings, and at pH 8.0 and 10.0 in leaves of mature plants. Seven-week-old plants were transferred to a hydroponic system and treated with different concentrations of Cd2+ or Cu2+ [in mM]: 0, 5, 25, 50, 100 for 7 days. The lipoxygenase activities at pH 8.0 and 10.0 depended on the metal that was added to the nutrient solution. The main change in lipoxygenase activity was under Cd2+ stress at pH 8.0 and under Cu2+ excess at pH 10.0.
Address and Contact Information Department of Plant Physiology, Maria Skłodowska-Curie University, Akademicka 19, 20-033 Lublin, Poland
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Volume 8 (2003) pp 285-296
Title VISUAL AND ARCHAEAL RHODOPSINS: SIMILARITIES, DIFFERENCES AND CONTROVERSY
Authors Krzysztof Bryl
Abstract Rhodopsins are currently known to belong to two distinct protein families. The visual rhodopsins, found in eyes throughout the animal kingdom, are photosensory pigments. Archaeal rhodopsins, found in extreme halophiles, function as light-driven proton pumps (bacteriorhodopsins), chloride ion pumps (halorhodopsins), or photosensory receptors (sensory rhodopsins). Light absorption by rhodopsins triggers their characteristic photoconversion extending into the (milli)second time range. There are three main paradigms of rhodopsins photoconversion. (1) Initiation of the trans-cis isomerization is the very primary consequence of light absorption. (2) Rhodopsins store light energy via the charge-separation mechanism (the charge of Schiff base is separated from its counterion). (3) Full trans-cis isomerization of the chromophore is a prerequisite for the full biological activity of rhodopsins. These paradigms will be questioned.
Address and Contact Information Department of Physics and Biophysics, University of Warmia and Mazury, 10-719 Olsztyn, Poland
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Volume 8 (2003) pp 297-303
Title HISTOLOGICAL PICTURES OF MUSCLES AND AN EVALUATION OF CELLULAR INFILTRATIONS IN HUMAN POLYMYOSITIS/DERMATOMYOSITIS, AS COMPARED TO THE FINDINGS IN EXPERIMENTAL GUINEA PIG MYOSITIS
Authors Hanna Gendek-Kubiak* and Ewa G. Gendek
Abstract We tested whether intramuscular injections of dermatomyositis (DM) patients' sera into guinea pig muscles can be used to transfer myositic alterations to these animals. Additionally, similar tests were performed using neoplastic patients' sera and sera from non-neoplastic, non-myositic patients. The DM patients' sera induced idiopathic inflammatory myopathy (IIM) type histological changes in muscle fibres in guinea pig quadriceps muscles, which were especially evident 72 h after sera injections. Immunohistochemical stainings of myositic guinea pig muscles were done for guinea pig pan-T-cells, monocytes/macrophages, the neuronal marker-protein gene product 9.5 (PGP 9.5) and protein S-100. Our studies proved that the factor(s) responsible for the appearance of characteristic alterations in diseased muscles during the course of DM is/are present in patient sera.
Address and Contact Information Department of Cytophysiology, Histology and Embryology, Medical University of Łódź, Narutowicza 60, 90-136 Łódź, Poland
* Corresponding author: E-mail: h_kubiak@hotmail.com
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Volume 8 (2003) pp 305-310
Title THE OPTIMAL EUKARYOTIC SIGNAL FOR TRANSLATION INITIATION FROM NON-AUG CODONS, PRESENT UPSTREAM OF BACTERIOPHAGE l P CISTRON, IS INACTIVE IN Escherichia Coli
Authors Borys Wróbel1,*, Bartosz Słomiński2 and Grzegorz Węgrzyn1,2
Abstract Expression of the replication genes of bacteriophage l, O and P, is believed to be translationally coupled. However, it was previously noted that, under conditions of amino acid starvation, when O is not synthesized, P continues to be expressed at a relatively high level. The results presented in this report, contrary to the previously presented hypothesis, suggest that an AGACUGGAU sequence (an optimal context for translation initiation from non- AUG codons in eukaryotes, and present upstream the P cistron) is inactive in Escherichia coli. Comparative sequence analysis confirms that such a signal is unlikely to be important for P synthesis. Instead, a weak Shine-Dalgarno sequence may be present upstream the P cistron, and be active in the absence of O gene expression.
Address and Contact Information 1Institute of Oceanology, Polish Academy of Sciences, Św. Wojciecha 5, 81-347 Gdynia, Poland,
2Department of Molecular Biology, University of Gdańsk, Kładki 24, 80-822 Gdańsk, Poland
*Corresponding author, Phone: (+48 58) 301 2241 ext. 377, Fax: (+48 58) 301 0072, E-mail: wrobel@biotech.univ.gda.pl
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Volume 8 (2003) pp 311-315
Title TTHERE IS NO EVIDENCE FOR THE EXISTENCE OF COMPLEX FORMATION BETWEEN DOXORUBICIN AND GLUTATHIONE
Authors Małgorzata Marszałek1, Małgorzata Bartosz2 and Grzegorz Bartosz1, 3
Abstract Doxorubicin is co-transported with glutathione by several multidrug resistance proteins (MRPs). In order to check whether weak non-covalent aggregates between doxorubicin and glutathione can be formed, which might be substrates for the transporter, the effect of glutathione on the partition coefficient of doxorubicin was studied. No evidence of an effect of glutathione (at levels up to 20 mM) on the partition coefficient of doxorubicin was found in the pH range of 4.0-7.4. These results indicate that non-covalent doxorubicin-glutathione complexes do not form.
Address and Contact Information 1Department of Molecular Biophysics, University of Łódź, Banacha 12/16, 90- 237 Łódź, Poland,
2Department of Physical and Health Education, University of Łódź, Rudzka 56, 93-423 Łódź, Poland and
3Department of Biochemistry and Cell Biology, University of Rzeszów, Rejtana 16A, 35-959 Rzeszów, Poland
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Volume 8 (2003) pp 317-331
Title THE ROLE OF STAT5 PROTEINS IN THE REGULATION OF NORMAL HEMATOPOIESIS IN A CORD BLOOD MODEL
Authors Magdalena Baśkiewicz-Masiuk1*, Marek Masiuk2, Ryszard Czajka3 and Bogusław Machaliński1*
Abstract The signal transducers and activators of transcription-STAT5A and STAT5B-are responsible for the control of proliferation, differentiation and apoptosis, via their effect on gene expression. They are activated by the binding of many cytokines, growth factors and hormones to their receptors on the cell surface. Many of these cytokines regulate hematopoietic cell development; therefore, STAT5 proteins are suggested to play an important role in hematopoiesis. There are numerous contradictory reports available in the literature on the role of STAT5 in normal hematopoietic cell development; hence, the question of the real function of STAT5 proteins clearly requires further studies. The aim of our study was to evaluate the role of STAT5 in normal hematopoiesis using oligodeoxynucleotide (ODN) strategy against STAT5 mRNA. We employed the RT-PCR method to study STAT5 mRNA expression in cells after their incubation with ODNs. We analyzed the effect of blocking STAT5 proteins on the viability and clonogenecity of the CFU-GM (Colony Forming Unit of Granulocyte-Macrophages) and the BFU-E (Burst Forming Unit of Erythrocytes) obtained from human cord blood (CB). The clonogenic growth of the cells was assessed in methylcellulose cultures according to the type of oligodeoxynucleotides. We also attempted to estimate the level of apoptosis induced in cord blood mononuclear and CD34+ cells by employing different assays: i) Annexin V staining using flow cytometry (FACSCalibur); ii) terminal deoxynucleotidyltransferase dUTP nick end labeling (TUNEL); iii) analysis of Bax and Bcl-XL gene expression by RT-PCR. Perturbation of STAT5 expression with antisense oligodeoxynucleotides had no impact on the viability, clonogenecity and apoptosis of CB hematopoietic cells. Our results showed that STAT5 proteins do not play a significant role in the regulation of proliferation of normal hematopoietic cells derived from cord blood.
Address and Contact Information 1Department of General Pathology, 2Department of Pathology, 3Clinic of Obstetrics, Pomeranian Medical University, Al. Powstańców Wlkp.72, 70-111 Szczecin, Poland
* Corresponding authors: Magdalena Baśkiewicz-Masiuk and Bogusław Machaliński, E-mail: poziomka@med.pam.szczecin.pl; machalin@sci.pam.szczecin.pl
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Volume 8 (2003) pp 333-336
Title THE EFFECT OF MELATONIN ON ANTIOXIDANT ENZYMES IN HUMAN DIABETIC SKIN FIBROBLASTS
Authors Ewa Kilańczyk and Maria Bryszewska*
Abstract Melatonin plays several important physiological functions in mammals, such as immune enhancement and regulation of dark-light signal transduction. Melatonin is also known to be an endogenous free radical scavenger and an efficient antioxidant. It detoxifies a variety of free radicals and reactive oxygen intermediates, including the hydroxyl radical, singlet oxygen and nitric oxide. These radicals participate in many diseases, for example diabetes. This study determined the effect of melatonin on the antioxidant enzymes: superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx), and the level of glutathione (GSH) in human diabetic (C2 line) skin fibroblasts. Confluent monolayers of control (S2 line) and diabetic (C2 line) skin fibroblasts were incubated with different concentrations of melatonin: 10, 50, 100 and 1000 mmol/l at 37oC for 24 h. Next, the GSH level and SOD, CAT and GPx activities were measured colorimetrically. The activities of the antioxidant enzymes and the GSH level were lower in diabetic skin fibroblasts than in the control S2 line. Concentrations of melatonin of 100 and 1000 mmol/l caused a significant increase in the enzymes' activities and GSH level.
Address and Contact Information Department of General Biophysics, University of Łódź, Banacha 12/16, 90-237 Łódź, Poland
*Corresponding author: Phone/fax: +48 42 635 44 74, E-mail: marbrys@biol.uni.lodz.pl
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Volume 8 (2003) pp 337-342
Title THE INFLUENCE OF METMYOGLOBIN AND FERRYLMYOGLOBIN ON THE HUMAN ERYTHROCYTE MEMBRANE
Authors Małgorzata Sztiller* and Mieczysław Puchała
Abstract Preliminary experiments revealed that ferrylmyoglobin decayed more slowly in the absence than in the presence of intact erythrocytes and erythrocyte membranes. This suggested the existence of interactions between FerrylMb and the erythrocyte membrane. Subsequent studies examined the influence of FerrylMb on the membrane of intact erythrocytes and on isolated erythrocyte membranes. The incubation of intact erythrocytes with FerrylMb did not influence their osmotic fragility or the fluidity of their membranes; the level of peroxidation of the membrane lipids increased only slightly( there was only a slight increase in the level of membrane lipid peroxidation). The activity of acetylcholinesterase significantly increased after 15 minutes of incubation, whereas longer incubation did not lead to any changes in the activity of this enzyme. The incubation of isolated erythrocyte membranes with FerrylMb resulted in an increase in their fluidity and a significant rise in the level of lipid peroxidation.
Address and Contact Information University of Łódź, Department of Molecular Biophysics, Banacha 12/16, Łódź, Poland
* Corresponding author
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Volume 8 (2003) pp 343-351
Title THE ANTHER-CULTURE RESPONSE OF TRITICALE LINE X TESTER PROGENIES
Authors Karol Marciniak1, Zygmunt Kaczmarek2*, Tadeusz Adamski2 and Maria Surma2
Abstract Seven triticale cultivars (Ampiac, Aubrac, Trinidad, Ticino, Lamberto, Pronto and Prado) and their F1 hybrids obtained after crossing in a line x tester scheme were examined with respect to their androgenetic effectiveness. The embryo induction rate (number of embryos per 100 anthers), green plant regeneration rate (number of green plantlets per 100 embryos), plant yield (number of green and albino plantlets per 100 anthers) and green plant yield (number of green plantlets per 100 anthers) were assessed. The multivariate and univariate effects of general (GCA) and specific (SCA) combining abilities for the studied traits were estimated and tested. Significant differences between the genotypes were found for individual traits as well as for all the traits treated jointly. Hybrids generally showed a better response in anther culture than their parental genotypes. Heterosis effects were observed in some hybrids for embryo induction rate and green plant yield. GCA and SCA variances were significant and a dominance of the GCA over the SCA variation was found. Among the examined cultivars, Ticino and Pronto were characterised by positive and significant GCA for embryo induction and green plant yield, and these cultivars may be recommended for the improvement of anther culture responsiveness in triticale.
Address and Contact Information 1DANKO Plant Breeding, Choryń, Poland,
2Institute of Plant Genetics, Polish Academy of Sciences, Strzeszyńska 34, 60-479 Poznań, Poland
*Corresponding author, E-mail: zkac@igr.poznan.pl
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Volume 8 (2003) pp 353-362
Title THE PRION PEPTIDE FORMS ION CHANNELS IN PLANAR LIPID BILAYERS
Authors Vladimir Berest1, Marcin Rutkowski2, Krzysztof Rolka3, Anna Łęgowska3 , Grażyna Dębska1, Dariusz Stępkowski2 and Adam Szewczyk1*
Abstract One of the hypotheses concerning the pathogenic properties of the prion protein considers its influence on cellular ion homeostasis. Using the lipid bilayer technique, the influence of prion-derived peptides on the lipid bilayer conductance was characterized. To evaluate the physiological significance and possible pathological functions of the peptides, their effect on the membrane potential and respiration rate of hippocampal mitochondria was also studied. We used a peptide bearing the human prion protein sequence YSNQNNF (PrP [169- 175]), and peptide SSQNNF (PrP [170-175]) bearing a naturally-occurring mutation in position 171 [N®S] linked to schizoaffective diseases in humans (Samaia, H.B., Mari, J.J., Vallada, H.P., Moura R.P., Simpson A.J.G., Brentani R.R. A prion-linked psychiatric disorder. Nature 390 (1997) 241). In this report, we show that PrP [170-175] N171S increases the conductance of planar lipid bilayers. Based on the conductance of single channel currents recorded in 500/500 mM KCl (cis/trans), we found a single channel conductance of 8 to 26 pS. The native prion peptide PrP [169-175] does not form ion channels in the lipid bilayer. Neither of the peptides significantly changed the membrane potential or respiration rate of isolated rat hippocampal mitochondria. We propose a possible mechanism for channel formation by aggregation of the prion-derived peptide.
Address and Contact Information 1Laboratory of Intracellular Ion Channels and 2Department of Muscle Biochemistry, Nencki Institute of Experimental Biology, Polish Academy of Sciences, Pasteura 3, 02-093 Warszawa, Poland,
3Faculty of Chemistry, University of Gdańsk, Sobieskiego 18, 80-952 Gdańsk, Poland
* Corresponding author: Tel: (4822) 6598571 ext. 269, Fax: (4822) 8225342, E-mail: adam@nencki.gov.pl
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Volume 8 (2003) pp 363-374
Title THE USE OF IMMOBILIZED METHYLCHYMOTRYPSIN FOR THE PURIFICATION OF HUMAN AND SHEEP ALPHA-1-PROTEINASE INHIBITOR (a1-PI)
Authors Joanna Grybel and Tadeusz Wilusz
Abstract a1-proteinase inhibitor was isolated from albumin fractions of human and sheep plasma using a new method of purification using affinity chromatography on immobilized methylchymotrypsin in the presence of 5 M NaCl. The inhibitor was finally polished to homogenity either by chromatography on a Mono Q or a Sephacryl S-200 HR column. The presented method makes it possible to recover a1-proteinase inhibitor which has been added to cow milk.
Address and Contact Information Institute of Biochemistry and Molecular Biology, University of Wrocław, Tamka 2, 50-137 Wrocław, Poland
Corresponding authors-Fax: (+48) 71-3752-608; E-mail: joanna@bf.uni.wroc.pl, wilusz@bf.uni.wroc.pl
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Volume 8 (2003) pp 375-381
Title AFLP MARKER POLYMORPHISM IN CUCUMBER (Cucumis sativus L.) NEAR ISOGENIC LINES DIFFERING IN SEX EXPRESSION
Authors Justyna Witkowicz, Ewa Urbańczyk-Wochniak and Zbigniew Przybecki
Abstract The AFLP technique was used to evaluate the level of polymorphism between two pairs of isogenic cucumber (Cucumis sativus L.) lines (NIL) differing in flower sex expression. The BSA techniques were also applied to find molecular markers linked to sex determination genes (dominant alleles) in those cucumber lines. Sex determination in cucumber is controlled by three main loci F, M and Gy. The interaction of these loci is responsible for the formation of various phenotypes of flowers in respect to sex in analyzed lines.: a female line 2gg with a ff/MM/gygy genotype, isogenic to a monoecious line B10 (genotype ff/MM/GyGy), and a female line Gy3 with a FF/MM/GyGy genotype, isogenic to a hermaphroditic line HGy3 (genotype FF/mm/GyGy). Using 56 combinations of AFLP primers, used for the analysis of lines 2gg and B10, gave 3794 bands, of which 155 (4.1%) were polymorphic. Ten bands distinguished gynoecious and monoecious bulks appearing at the same time in the appropriate parent; they are believed to be linked to the Gy locus. The isogenic lines Gy3 and HGy3 showed a higher level of polymorphism (14.2%). In this case, 55 combinations of primers gave 2996 reaction products, of which 430 showed variation. Twenty bands occurred in one bulk and in one parent, so they are probably associated with the M locus. Using the AFLP technique, the isogenicity of the lines was evaluated. The level of polymorphism (per pair of primer) between lines 2gg and B10 is 0.072% and is four times lower than that between the Gy3 and HGy3 lines (0.27%). The differences in the isogenicity of the lines can result from the degree of their relatedness, which may reflect the way they were derived.
Address and Contact Information Department of Plant Genetics Breeding and Biotechnology, Warsaw Agricultural University, Nowoursynowska 166, 02-787 Warsaw, Poland
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Volume 8 (2003) pp 383-389
Title THE EFFECT OF THE PRESENCE OF CROWN ETHER ON ION TRANSPORT ACROSS THE LIPID BILAYER
Authors Monika Naumowicz1, Aneta D. Petelska1 and Zbigniew A. Figaszewski1,2*
Abstract We studied the electric properties of phosphatidylcholine bilayers modified with crown ether (dibenzo[18] crown-6). The studies were carried out for various crown ether concentrations in forming solutions and various potassium ion concentrations in electrolyte solutions. The presence of crown ether in the membrane influences the membrane's impedance; there is a reduction in its resistivity, a decrease in its resistance of phase transfer and an increase in its capacity of phase transfer with an increase in crown ether concentration in the bilayer and in K+ ion concentration in the electrolyte solution.
Address and Contact Information 1Institute of Chemistry, University of Białystok, Al. J. Piłsudskiego 11/4, 15-443 Białystok, Poland,
2Laboratory of Electrochemical Power Sources, Faculty
* Corresponding author, Fax: +4885 745 75 81, E-mail: elchem@uwb.edu.pl
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Volume 8 (2003) pp 391-413
Title DNA DAMAGE CAUSED BY LIPID PEROXIDATION PRODUCTS
Authors Wojciech Łuczaj and Elżbieta Skrzydlewska
Abstract Lipid peroxidation is a process involving the oxidation of polyunsaturated fatty acids (PUFAs), which are basic components of biological membranes. Reactive electrophilic compounds are formed during lipid peroxidation, mainly a,b-unsaturated aldehydes. These compounds yield a number of adducts with DNA. Among them, propeno and substituted propano adducts of deoxyguanosine with malondialdehyde (MDA), acrolein, crotonaldehyde and etheno adducts, resulting from the reactions of DNA bases with epoxy aldehydes, are a very important group of adducts. The epoxy aldehydes are more reactive towards DNA than the parent unsaturated aldehydes. The compounds resulting from lipid peroxidation mostly react with DNA showing both genotoxic and mutagenic action; among them, 4- hydroxynonenal is the most genotoxic, while MDA is the most mutagenic. DNA damage caused by the adducts of lipid peroxidation products with DNA can be removed by the repairing action of glycosylases. The formed adducts have been hitherto analyzed using the IPPA (Imunopurification-32P-postlabelling assay) method and via gas chromatography/electron capture negtive chemical ionization/mass spectrometry (GC/EC NCI/MS). A combination of liquid chromatography with electrospray tandem mass spectrometry (LC/ES-MSMS) with labelled inner standard has mainly been used in recent years.
Address and Contact Information Department of Analytical Chemistry, Medical Academy of Białystok, Mickiewicza 2A, P.O. Box 14, 15-230 Białystok 8, Poland
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Volume 8 (2003) pp 415-419
Title A COMPARISON OF THE TOTAL ANTIOXIDANT CAPACITY OF SOME HUMAN BODY FLUIDS
Authors Anna Ziobro1* and Grzegorz Bartosz1,2
Abstract The Total Antioxidant Capacity of several human fluids was compared and the following sequence of TAC values was found: urine > saliva > blood plasma > milk ≈ˆ amniotic fluid >> sweat. Lower TAC values were found for the saliva of smokers than for that of non-smokers. Drinking of a cup of instant coffee increased the hydrogen peroxide content of urine but did not decrease the TAC of urine.
Address and Contact Information 1Department of Biochemistry and Cell Biology, University of Rzeszów, Rejtana 16A, 35-959 Rzeszów, Poland,
2Department of Molecular Biophysics, University of Łódź, Banacha 12/16, 90-237 Łódź, Poland
* This work was done as the Master's (M. Sc.) thesis of Anna Ziobro in the Institute of Biology and Environment Protection, University of Rzeszów.
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Volume 8 (2003) pp 421-438
Title THE ISOLATION OF cDNA CLONES FROM CUCUMBER (Cucumis sativus L.) FLORAL BUDS COMING FROM PLANTS DIFFERING IN SEX
Authors Zbigniew Przybecki*, Magdalena Ewa Kowalczyk, Ewa Siedlecka, Ewa Urabańczyk-Wochniak and Stefan Malepszy
Abstract In this study, we found flower cDNA clones which may be connected with the development of flower sex in cucumber. Two pairs of nearly-isogenic lines: gynoecious GY3 (FFMMGG) versus hermaphrodite HGY3 (FFmmGG) and monoecious B10 (ffMMGG) versus gynoecious 2gg (ffMMgg) were used for clone isolation. To obtain differentially-expressed clones, we applied the differential screening method. 454 clones from GY3 and 478 from B10 cDNA libraries were isolated. The results of RFLP analysis with 56 cDNA clones showed no clones which cosegregated with sex in cucumber. The 28 cDNA B10 and 33 cDNA GY3 clones isolated using the differential screening method were sequenced. Some of them seem to may play a role in cell differentiation or flower development. Among the 61 identified clones, 14 show high homology to plant proteins, although of unknown function. 11 show high homology to known proteins, and the possible function of some of them is discussed. For 3 clones, no significant similarity was found. The 31 clones displayed high homology to plant cDNA in EST database. The patterns of expression of five differential cDNA clones, 35GY3, 216GY3, 47GY3, 100B10 and 157B10, were analyzed in cucumber flower buds using in situ RT-PCR. The most interesting clone is 35GY3, because of its possible role in the inhibition of the development of male specific elements in the female cucumber flower
Address and Contact Information Department of Plant Genetics, Breeding and Biotechnology, Faculty of Horticulture, Warsaw Agricultural University, Nowoursynowska 166, 02-787 Warsaw, Poland
* Corresponding author, E-mail: przybecki@alpha.sggw.waw.pl
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Volume 8 (2003) pp 439-454
Title A KINETICS STUDY OF PIG ERYTHROCYTE HEMOLYSIS INDUCED BY POLYENE ANTIBIOTICS
Authors Agnieszka Knopik-Skrocka*, Józef Bielawski**, Marta Głąb, Agnieszka Klafaczyńska and Monika Wulkiewicz
Abstract The kinetics of the hemolysis induced by filipin is of the damage type, indicating the formation of large nonselective perforations of erythrocyte membranes. The process is relatively independent of the ionic composition of the incubation medium, and the differences between the hemolysis induced by filipin in pig and human erythrocytes are not significant. In a sucrose medium, filipin-induced hemolysis is inhibited in humans, whereas it is stimulated in pig erythrocytes. It is suggested that low ionic strength is the reason for the different modifications of complexation of filipin in pig and human erythrocyte membranes in a sucrose medium. The kinetics of the hemolysis induced in pig erythrocytes by amphotericin B and nystatin is of the permeability type, indicating the formation of selective channels in erythrocyte membranes and colloid osmotic hemolysis. The rate of the hemolysis, which is high in a KCl medium, is decreased in all the other media tested (CaCl2, MgCl2, potassium phosphate buffer, K2SO4, sucrose), although there are no changes in the kinetics of hemolysis. The results are interpreted as the formation of highly selective channels at a low concentration of the antibiotics. At increasing concentrations, channels of decreasing selectivity occur. The resistances of pig erythrocytes to amphotericin B and nystatin are lower than those of human erythrocytes.
Address and Contact Information Department of Cytology and Histology, A. Mickiewicz University, Fredry 10, 61-701 Poznań, Poland
* E-mail:askro@amu.edu.pl, ** E-mail: bielj@amu.edu.pl
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Volume 8 (2003) pp 455-460
Title THE EFFECT OF PEROXYNITRITE AND SOME ANTIOXIDANTS ON THE RATE OF OSMOTIC HEMOLYSIS OF BOVINE ERYTHROCYTES
Authors Anna Wróbel1*, Beata Łukaszyńska1 and Jadwiga Kędzierska2
Abstract Bovine erythrocytes treated with peroxynitrite (ONOO-), a cytotoxic species formed in vivo via the reaction of nitric oxide (NO·) and the superoxide anion (O ), show an increased rate of hemolysis on sudden osmotic stress. The increase in the rate was peroxynitrite concentration dependent. In the presence of some antioxidants (uric acid, ascorbic acid, glutathione, melatonin and albumin), this effect was significantly lower, with ascorbic acid as the most efficient antioxidant.
Address and Contact Information 1Institute of Physics, Wrocław University of Technology, Wybrzeże Wyspiańskiego 27, 50-370 Wrocław, Poland,
2Faculty of Chemistry, University of Wrocław, F. Joliot-Curie 14, 50-383 Wrocław, Poland
* Corresponding author: E-mail: Anna.Wrobel@pwr.wroc.pl
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Volume 8 (2003) pp 461-470
Title PROPHYLACTIC EFFECT OF MELATONIN IN REDUCING LEADINDUCED NEUROTOXICITY IN THE RAT
Authors Gamal H. El-Sokkary1, Esam S. Kamel2 and Russel J. Reiter3*
Abstract Oxidative stress is a likely molecular mechanism in lead neurotoxicity. Considering the antioxidant properties of melatonin, this study investigated the neuroprotective potential of melatonin in the hippocampus and corpus striatum of rats treated with lead. Three groups of male rats (control, lead acetate-treated [100 mg/kg], and lead acetate plus melatonin [10 mg/kg] for 21 consecutive days) were used. Levels of products of lipid peroxidation (LPO), glutathione (GSH) and superoxide dismutase (SOD) activity were measured in brain homogenates. Histological changes in the pyramidal cells of the hippocampus and the putamen of the corpus striatum were examined. The results documented increased LPO and decreased GSH and SOD activity in the brain homogenates of lead-treated rats. Histological observations revealed severe damage and a reduction in neuronal density in the hippocampus and corpus striatum. When melatonin was given to lead-treated rats, it almost completely attenuated the increase in LPO products and restored GSH levels and SOD activity. Also, the morphological damage was reduced and neuronal density was restored by melatonin. Considering the ease with which melatonin enters the brain, these results, along with previous observations, suggest that melatonin may be useful in combating free radical-induced neuronal injury that is a result of lead toxicity.
Address and Contact Information 1Department of Zoology, Faculty of Science, Assiut University, Assiut, Egypt,
2Department of Anatomy, Faculty of Medicine, South Valley University, South Valley, Egypt, 3Department of Cellular and Structural Biology, University of Texas Health Science Center, 7703 Floyd Curl Drive, San Antonio,
* Corresponding author: Phone: 210/567-3859; Fax: 210/567-6948; E-mail: reiter@uthscsa.edu TX 78229-3900, USA
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Volume 8 (2003) pp 471-486
Title THE TUMORIGENIC POTENTIAL OF HUMAN CX-1 COLON ADENOCARCINOMA CELLS DEPENDS ON CARCINOEMBRYONIC ANTIGEN (CEACAM5) EXPRESSION
Authors Dagmara Baczyńska1, Joanna Wietrzyk2, Janusz Madej3, Anna Krop-Wątorek1, Anna Dąbrowska1, Katarzyna Widerak1, Adam Opolski2 and Maciej Ugorski1,4
Abstract It was shown that CEACAM5 can mediate cell-cell adhesion through homotypic and heterotypic interactions; however, its role in the expression of the malignant phenotype remains obscure. To study whether the formation of both primary tumors and metastases is directly related to the presence or absence of CEACAM5, we applied the antisense RNA strategy. By transfecting human CX- 1.1 colon carcinoma cells with CEACAM5 antisense-expressing vector or with the vector itself, cell variants with a highly decreased expression of CEACAM5 were obtained. Profound differences in proliferative abilities among parental and obtained subclones of CX-1.1 cells were revealed when cells were implanted subcutaneously into nude mice. In contrast to their highly tumorigenic parental CX-1.1 cells (with high expression of membrane-bound and secreted CEACAM5), two subclones (3E and AS6Q) with substantially decreased expression of membrane-bound and secreted CEA showed a considerably diminished growth rate. Even more striking results were obtained with AS8Q cells, producing a residual amount of this glycoprotein. However, 3B cells (producing a large amount of secreted CEACAM5) did not differ significantly in their tumorigenic properties from CX-1.1 cells. Our experiments performed in nu/nu mice suggest that CEACAM5 supports the growth of primary tumors, but is not involved in the formation of metastases by colon cancer cells.
Address and Contact Information 1Departments of Immunochemistry and 2Tumor Immunology, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Rudolfa Weigla 12, 53-114 Wrocław, Poland,
3Department of Pathology, Faculty of Veterinary Medicine, Agriculture University, Cypriana Norwida 31, 50-375 Wrocław, Poland
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