The transfer of bacterial biotin genes to Escherichia coli K-12 cells by transfection with lambda bio phage DNA.

The transfection of biotin genes to biotin negative Escherichia coli K-12 cells with donor DNA isolated from transducing lambda bio phages was examined. The transfection of the cells was performed in the presence of bovine serum albumin and sucrose (Osowiecki and Skalińska, 1974).     

Nuclear distribution pattern of tumour-associated nonhistone protein of mol. wt 48,000.

• 1.1. As a further step toward characterizing nonhistone protein of mol. wt 48,000 which was found to be much more abundant in animal tumour cells than in normal ones [Krajewska W.M., Lipinska A., Marszatek M., Kiliańska Z., Wojtkowiak Z. and Kłyszejko-Stefanowicz L. Cell. Biochem. Funct. 8, 79–89 (1990)] its intranuclear localization in hamster liver and Kirkman-Robbins hepatoma was studied. The protein was identified by immunoblotting technique in the presence of antibodies against polypeptide of mol. wt about 48,000 from Kirkman-Robbins hepatoma.
• 2.2. Distribution of antigen with mol. wt of 48,000 in nuclear fractions representing different levels of nuclear material organization i.e. in nucleoli, nuclease-sensitive and nuclease-resistant fractions, and extensive nuclease digestion products separated by size on Bio-Gel A-50m, implied the structural role of this component.
• 3.3. Fractionation of endogenously digested nuclei into low salt extract, high salt extract and nuclear matrix revealed that in normal liver the antigen studied is associated with nuclear matrix while in hepatoma this component appeared in high salt extract.
• 4.4. These results suggest that polypeptide with mol. wt of 48,000 is a shuttling protein which may be involved in reorganization of nuclear matrix during neoplastic transformation.

     

Olminska and colleagues’ study, “Conversion of the helical tomotherapy plans to the step-and-shoot IMRT plans for patients with hip prosthesis during radiotherapy for prostate cancer” was interesting to read

Olmińska and colleagues’ study, Olmińska et al. (2016) was interesting to read [1]. While prasining the authors for their great work, I want to emphasize e few points.In the recent years, with the development of new device technology, Intensity Modulated Radiotherapy (IMRT) and complex treatment modalities such as stereotactic radiosurgery and helical tomotherapy were started to be implemented. Thus, due to increased local control of tumor growth and reduction of dose received by surrounding critical organs, serious complications were avoided. In this new treatment modality, while calculating appropriate dose, all the parameters such as patient anatomy and characteristics of radiation should be taken into account. Besides, during conformal radiotherapy, if hip prosthesis is located around or in the clinical target volume (CTV), type, thickness and density of biomaterial should be considered to avoid dose differences.     

Aspartate and asparagine tRNA genes in wheat mitochondrial DNA: a cautionary note on the isolation of tRNA genes from plants.

We have identified genes encoding a "native" tRNA(Asp) (trnD-GTC) and a "chloroplast-like" tRNA(Asn) (trnN-GTT) on opposite strands and 633 bp apart within a sequenced 1640 bp RsaI restriction fragment of wheat mtDNA. The trnD gene has been found previously at a different location in wheat mtDNA (P.B.M. Joyce et al. (1988) Piant Mol. Biol. 11, 833-843); the duplicate copies of this gene are identical within the coding and immediate flanking regions (9 bp downstream and at least 68 bp upstream), after which obvious sequence similarity abruptly disappears. The trnN gene is identical to its homolog in maize ctDNA; continuation of sequence similarity beyond the coding region suggests that this gene originated as promiscuous ctDNA that is now part of the wheat mitochondrial genome. In the course of this work, we have encountered some unexpected similarities between tRNA gene regions from wheat mitochondria and other sources. Detailed analysis of these similarities leads us to suggest that trnN genes reportedly from petunia nuclear DNA (N. Bawnik et al. (1983) Nucleic Acids Res. 11, 1117-1122) and lupine mtDNA (B. Karpińska and H. Augustyniak (1988) Nucleic Acids Res. 16, 6239) are, in fact, from petunia mtDNA and lupine ctDNA, respectively, whereas a putative wheat nuclear tRNA(Ser) (trnS-TGA) gene (Z. Szwekowska-Kulińska et al. (1989) Gene 77, 163-167) is actually from wheat mtDNA. In these instances, it seems probable that the DNA samples used for cloning contained trace amounts of DNA from another sub-cellular compartment, leading to the inadvertent selection of spurious clones.     

中国水华蓝藻的新记录属——拟浮丝藻属(Planktothricoides)

在近年来对水华蓝藻的调查中,确定了我国一水华蓝藻新记录属——拟浮丝藻属Planktothricoides(WoIoszyńska) Suda et Watanabe 2002。文中对该属及该属一个新记录种的主要形态学特征进行了描述,并对其相近属浮丝藻属Planktothrix进行了形态学比较研究。     

Factors affecting oxygen-induced changes in the activity of CTP-dependent lipid kinases in yeast

Previous work from this Laboratory (Szkopińska et al., 1988, Arch. Biochem. Biophys., 266, 124-131) indicated that CTP is a phosphate donor for the synthesis of phosphatidic acid and dolichyl phosphates. The elucidation of the role of mitochondrial membranes and mitochondrial proteins (isolation of rho- mutant) as well as specific detergents and sterols has been the aim of this work.     

Dependence of mitochondrial oxidative phosphorylation on activity of the adenine nucleotide translocase

The coupled reactions of electron transport and ATP synthesis for the first two sites of mitochondrial oxidative phosphorylation have been previously reported to be near equilibrium in isolated respiring pigeon heart (Erecińska, M., Veech, R. L., and Wilson, D. F. (1974) Arch. Biochem. Biophys. 160, 412-421) and rat liver mitochondria (Forman, N. G., and Wilson, D. F. (1982) J. Biol. Chem. 257, 12908-12915). Measurements are presented in this paper which demonstrate that the same relationship exists for both forward and reverse electron transport in rat heart mitochondria. This conclusion implies that adenine nucleotide translocation, a partial reaction of the system, is also near equilibrium, contrasting with proposals that the translocase is rate-limiting for oxidative phosphorylation. To resolve this controversy, the respiratory rates of suspensions of isolated rat liver and rat heart mitochondria were controlled by varying either the added [ATP]/[ADP][Pi] ratios ratios or [ADP] (by varying hexokinase in a regenerating system). Titrations with carboxyatractyloside, a high affinity inhibitor of the translocase which is noncompetitive with ADP, were carried out to assess the dependence of the respiratory rate on translocase activity. Plots of respiratory rate versus [carboxyatractyloside] were all strongly sigmoidal. In liver mitochondria, 40%-70% and in heart mitochondria 66% of the sites could be blocked with carboxyatractyloside before a 10% decrease in the respiratory rate was observed. Further analysis showed that liver and heart mitochondria have translocase/cytochrome a ratios of 1.52 and 3.20, respectively, and that at 23 degrees C the maximal turnover numbers for the translocases were 65 s-1 and 23 s-1. In all states of controlled respiration (no added inhibitor), a substantial excess of translocase activity was present, suggesting that the translocase was not normally rate-limiting in oxidative phosphorylation.     

THE ULTRASTRUCTURE OF GLENODINIOPSIS STEINII (DINOPHYCEAE)

The freshwater dinoflagellate Glenodiniopsis steinii Wolsoszyńska was examined using computer-assisted three-dimensional reconstruction of serially sectioned cells observed with the transmission electron microscope and images from the scanning electron microscope. Vegetative cells contain ultrastructure typical of freshwater dinoflagellates including trichocysts, mitochondria, Golgi bodies, starch grains, and lipid bodies. The chloroplast is a single, multilobed structure, not multiple discoid chloroplasts as previously described. The “C” shape of the nucleus is apparently due in part to the size and location of the pusule.     

Studies on the morphology of colonies of bacilli of BCG-Poland substrain. II. Dissociation of BCG-Poland substrain on L?wenstein-Jensen egg medium.

The dissociation of colonies of the substrain BCG-Poland on L?wenstein-Jensen medium has been studied. It had been stated previously (Lipińska and Rzucid?o, 1974) that BCG-Poland substrain undergoes a process of dissociation which manifests itself in the appearance of smooth colonies alongside typical rough colonies. Attempts have been made to obtain pure culture of both colony types. Pure cultures composed of smooth or rough colonies respectively were obtained. The morphology and diameter of the smooth colonies depended on the size of the seed inoculum spread on the medium.     

Christensonella ecallosa sp. nov. (Orchidaceae,Maxillariinae) from Guyana and Brazil

A new species of Christensonella (Orchidaceae, Maxillariinae) is described and illustrated based on herbarium material. The most important characters of Christensonella ecallosa Lipińska & Szlach. are the ovate lip, with transversely elliptic middle lobe, minutely dentate margins and very obscure lip callus, which forms an elevated thick pad near the base. Taking into account the habit, it may be misidentified as C. uncata, but they are clearly distinguished by the lip shape. The taxonomic affiliations of the new species are briefly discussed.     

Genotoxicity of lead in lupin root cells as evaluated by the comet assay

This paper presents the results of a study on the influence of lead (Pb(+2)) on DNA integrity on plant cells. The study was performed on the root tips of lupin (Lupinus luteus cv. Juno) seedlings treated with two selected concentrations of Pb(NO3)2: 150 and 350 mg l(-1), which were found to inhibit root growth by 50% and 70%, respectively [Rucińska et al. Plant Physiol. Biochem. 37 (1999) 37187-37194]. Roots exposed to those external lead concentrations took up about 50 and 70 mg l(-1) Pb(+2) g(-1) fresh weight (FW) over 48 h of incubation. A dose-dependent increase in the degree of root injury was observed in the presence of both tested concentrations. The genotoxicity of lead in lupin root cells was analysed using a mild alkaline comet assay at pH 12.3, which allows the detection of single strand breaks. The quantity of the DNA fragments migrating away from the nuclear remnant (tail area) increased proportionally to the lead content inside the roots, and was positively correlated with the degree of root injury. At 150 mg l(-1) Pb(+2), a high frequency distribution of nuclei having large values of tail lengths and moments was observed. By contrast, the number of nuclei with minimum values of these parameters increased at 350 mg l(-1) Pb(+2). This data suggests that lead at low concentrations induces the formation of short, rapidly migrating DNA fragments, whereas at higher concentrations, lead probably causes other changes to DNA that result in slower DNA migration in the electric field.     

The relationship between chronotype and personality among patients with alcohol dependence syndrome: Pilot study

The study investigates the distribution of chronotypes among alcohol-dependent subjects and the relationship between personality and chronotype. Fifty-eight alcohol-dependent patients and 29 age-matched healthy controls were studied using Ogińska’s Chronotype Questionnaire (ChQ), Eysenck’s Personality Questionnaire – Revised (EPQ-R), Selzer’s Michigan Alcoholism Screening Test (MAST) and a sociodemographic status questionnaire designed by the authors. The alcohol-dependent patients tended to be morning type, based on the morningness–eveningness ChQ scale, with a weakly marked rhythm, based on the distinctness ChQ scale. Preference towards morningness was associated with older age, but no relation between chronotype and severity of alcohol dependence was found. A high amplitude of the rhythm was associated with higher neuroticism. Therefore, despite being in the minority, patients with a distinct circadian rhythm (i.e. with a high amplitude) are at greater risk of mood and anxiety disorders and hence should be given special consideration.     

Synthesis of Zn(II)-cloxacillin sodium complex and study of its interaction with calf thymus DNA

In this paper, a solid complex of cloxacillin sodium (CS) with Zn(II) was prepared by coprecipitation and characterized by UV, fluorescence, IR, and thermal spectra. Furthermore, the nature of the complex has been studied by ¹H-NMR and ¹³C-NMR spectroscopy. The influence of Zn(II) on the combination of CS and calf thymus DNA (CT DNA) was studied using fluorescence spectrophotometry, and the formation of binary CS-Zn(II) and CS-CT DNA complexes and ternary CS-Zn(II)-CT DNA complex was studied. The results show that the fluorescence intensity of CS can be quenched in the presence of Zn(II) or DNA. In the presence of Zn(II), the fluorescence quenching action of DNA on CS was strongly enhanced. Based on the fluorescence intensity, the formation constants of CS-Zn(II) and CS-CT DNA complexes were calculated, and the mechanism of interaction between CS, Zn(II), and DNA is discussed. Published in Russian in Biokhimiya, 2007, Vol. 72, No. 2, pp. 184–193.     

Enhanced Nasal Mucosal Delivery and Immunogenicity of Anti-Caries DNA Vaccine through Incorporation of Anionic Liposomes in Chitosan/DNA Complexes

The design of optimized nanoparticles offers a promising strategy to enable DNA vaccines to cross various physiological barriers for eliciting a specific and protective mucosal immunity via intranasal administration. Here, we reported a new designed nanoparticle system through incorporating anionic liposomes (AL) into chitosan/DNA (CS/DNA) complexes. With enhanced cellular uptake, the constructed AL/CS/DNA nanoparticles can deliver the anti-caries DNA vaccine pGJA-P/VAX into nasal mucosa. TEM results showed the AL/CS/DNA had a spherical structure. High DNA loading ability and effective DNA protection against nuclease were proved by gel electrophoresis. The surface charge of the AL/CS/DNA depended strongly on pH environment, enabling the intracellular release of loaded DNA via a pH-mediated manner. In comparison to the traditional CS/DNA system, our new design rendered a higher transfection efficiency and longer residence time of the AL/CS/DNA at nasal mucosal surface. These outstanding features enable the AL/CS/DNA to induce a significantly (p<0.01) higher level of secretory IgA (SIgA) than the CS/DNA in animal study, and a longer-term mucosal immunity. On the other hand, the AL/CS/DNA exhibited minimal cytotoxicity. These results suggest that the developed nanoparticles offer a potential platform for DNA vaccine packaging and delivery for more efficient elicitation of mucosal immunity.     

An interplay between the TOM complex and porin isoforms in the yeast Saccharomyces cerevisiae mitochondria.

The outer mitochondrial membrane of Saccharomyces cerevisiae contains two isoforms of mitochondrial porin, known also as the voltage-dependent anion channel. The isoform termed here porin1 displays channel-forming activity enabling metabolite transport whereas the second one, termed here porin2, does not form a channel and its function is still not clear. We have shown recently that in the absence of porin1, the channel within the protein import machinery (the TOM complex) is essential for metabolite transport across the outer membrane [Kmita and Budzińska, Biochim. Biophys. Acta 1509 (2000) 6044-6050]. Here, we report that the TOM complex channel may also serve as a supplementary pathway for metabolites in the presence of porin1 when the permeability of the latter is limited and the role of the TOM complex seems to increase when porin2 is depleted.     

Relationship of the intra- and extramitochondrial adenine nucleotide ratios during synthesis of phosphoenolpyruvate using extramitochondrial ATP

Mitochondria prepared from the livers of guinea pig, chicken, and pigeon all actively synthesize phosphoenolpyruvate from oxalacetate and GTP, utilizing phosphoenolpyruvate carboxykinase. It was previously shown (Wilson, D. F., Erecińska, M., and Schramm, V. L. (1983). J. Biol. Chem. 258, 10464-10473) that phosphoenolpyruvate carboxykinase is freely reversible and that, in conjunction with nucleoside diphosphate kinase and malate dehydrogenase, which are also present in the mitochondria, it can be used to measure the intramitochondrial [ATPfree]/[ADPfree]. In this study, synthesis of phosphoenolpyruvate by guinea pig liver mitochondria was studied under conditions for which the only source of GTP was extramitochondrial ATP via adenine nucleotide translocase and nucleoside diphosphate kinase (the mitochondria were treated with rotenone, oligomycin, uncoupler, and fluorocitrate). When the extramitochondrial [ATP]/[ADP] was greater than the intramitochondrial [ATPfree]/[ADPfree] calculated from the phosphoenolpyruvate carboxykinase reaction, there was net synthesis of phosphoenolpyruvate, but when it was less, there was net disappearance of phosphoenolpyruvate. Thus, the intramitochondrial [ATPfree]/[ADPfree] was equal to the extramitochondrial value at the point of reversal of the phosphoenolpyruvate carboxykinase reaction. This equality of the intra- and extramitochondrial adenine nucleotide ratios occurred with a measured mitochondrial membrane potential of approximately -36 mV, whereas in the previous experiments, equality was observed for conditions in which the measured membrane potential was -111 to -125 mV. Thus, adenine nucleotide translocation was not dependent on the transmembrane electrical potential and must, therefore, have occurred by electroneutral exchange.     

Low temperature, gibberellin and acid lipase activity in removal of apple seed dormancy

The activity of acid lipase and the level of gibberellin A₄ (GA₄) were determined in apple embryos excised from seeds after different time periods of stratification and subsequently cultured in darkness at 4°C or at 25°C. Enzyme activity and GA₄ content were higher at 4°C. Exogenous gibberellin stimulated lipase activity, while AMO-1618, an inhibitor of gibberellin biosynthesis, inhibited, to the same degree, both the enzyme activity and the GA₄ accumulation. The involvement of GA₄ and lipolytic enzymes in cold-mediated removal of embryonal dormancy has been discussed and compared with the role of these two factors in light-stimulated germination of dormant apple embryos, described earlier (Smoleńska and Lewak 1974).     

Electrotransformation of Streptococcus pyogenes with plasmid and linear DNA

Electrotransformation was used to introduce both plasmid and linear DNA into Streptococcus pyogenes. The method was optimized using strain NZ131, for which transformation frequencies up to 10(7) per micrograms of plasmid DNA were obtained. A linear fragment of DNA, containing the streptokinase gene (ska) in which an internal fragment had been replaced with an erythromycin resistance gene (erm), was transformed into strain NZ131 with a frequency of 10(3) per micrograms DNA. The introduction of linear DNA into S. pyogenes by electrotransformation should be useful for future genetic analyses as well as targeted gene replacement.     

Effect of modification of 6-[(aminoalkyl)amino]-7H-benzo[e]-perimidin-7-ones on their cytotoxic activity toward sensitive and multidrug resistant tumor cell lines. Synthesis and biological evaluation

Benzoperimidines, a novel group of antitumor anthracenedione analogues, are of interest due to their ability to overcome multidrug resistance of tumor cells (Stefańska, B., Dzieduszycka, M., Bontemps-Gracz, M. M., Borowski, E., Martelli, S., Supino, R., Pratesi, G., De Cesare, MA., Zunino, F., Ku?nierczyk, H., Radzikowski, Cz. J. Med. Chem. 1999, 42, 3494). Although the structural factor essential for exhibiting this desirable property is the presence in the molecule of a fused heterocyclic ring, the cytotoxicity against resistant cells is highly influenced by the nature and location of the substituents. A series of novel synthetic derivatives, comprising monohydroxylated benzoperimidines and 2-aminobenzoperimidines, allowed the establishment of an in vitro structure-activity relationship for a panel of leukemia sensitive, as well as P-gp dependent multidrug resistance (MDR) and multidrug resistance associated protein dependent resistance (MRP) resistant cell lines. The membrane affinity for the compounds has also been determined.     

Hepatoprotective effects of a concentrate and components of sake against galactosamine (GalN)-induced liver injury in mice

We investigated the hepatoprotective effects of a concentrate of sake (CS) and its components against D-galactosamine (GalN)-induced liver injury by measuring the plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in mice. CS significantly suppressed the GalN-induced elevation of ALT and AST activities. Each of four concentrated fractions extracted from sake (respectively consisting mainly of basic amino acids, neutral and acidic amino acids, organic acids and sugars) suppressed the GalN-induced elevation of ALT and AST activities. We focused on the sugar fraction containing glucose and ethyl alpha-D-glucoside (alpha-EG), which is a sake-specific sugar, as the major components and demonstrated that only alpha-EG showed significant suppression of the GalN-induced elevation of ALT and AST activities. We compared the effects of the alpha-EG analogues, methyl alpha-D-glucoside and ethyl beta-D-glucoside, on GalN-induced liver injury and confirmed that only alpha-EG significantly suppressed both the ALT and AST activities. Moreover, CS and alpha-EG suppressed the GalN-induced production of interleukin 6 (IL-6) and liver DNA fragmentation. Together these results show that CS and its component, alpha-EG, suppressed GalN-induced liver injury by inhibiting IL-6 production.