Release of mtDNA from mitochondria and activation of its replication in tissues of irradiated mice

A nessessary condition for normal functioning of mitochondria is the maintenance of certain numbers of intact mtDNA molecules. In the present study, we investigasted changes in the number of mtDNA copies in brain and spleen cells of mice subjected to irradiation. For the first time, we observed the irradiation-induced output of mtDNA fragments into brain and spleen cell cytosol. In the cytosol of these cells, examined in mice 5 h after 5 Gy irradiation, 1841 h.p. mtDNA fragments were detected able to persist for at 3 weeks. In addition, larger fragments of mtDNA (10,090 b.p.) were detected in the cytosol of brain cells of irradiated mice. The occurrence of mtDNA fragments in the cytosol of brain cells is accompanied with an increase in the number of mtDNA copies in the mitochondrial matrix. The induction of mtDNA replication in brain cells of irradiated animals may be considered as a compensatory reaction in response to mtDNA damage. A sharp decrease in the amount of mtDNA copies in the mitochondrial matrix of spleen cells on the first day after irradiation may be considered as apoptosis development. However, the compensatory reaction in brain cells was also noticed but in later terms.     

The effect of N-stearoylethanolamine on lipid composition of the metastases and conditionally normal lung tissue in mice with Lewis carcinoma

Influence of NSE on lipid composition of metastases and the neighbouring conditionally normal lung tissue in mice with Lewis carcinoma was investigated. The processes of peroxidation in investigated tissues were also studied. It was shown that under the influence of NSE the high level of antioxidant activity in the metastases was decreased, while in the neighbouring conditionally normal lung tissue the catalase activity was increased. The content of the thiobarbituric acid-reactive substances in comparison with animals which were not fed by NSE was decreased. The development of carcinoma was accompanied by significant decrease of cholesterol level and by the increase of unsaturated fatty acids esterified in membrane phospholipids in both the metastases and the neighbouring conditionally normal lung tissue. An analysis of the phospholipid spectra shows that under tumor growth in investigated tissues the high-level lysophosphatidylcholine (LPC) was observed. The content of phosphatidyl choline (PC), phosphatidyl ethanolamine (PE), phosphatidyl serine (PS) was found to be significantly lower than in the lung of intact animals. It was found that administration of NSE to tumor-bearing mice contributed to the increase of cholesterol level, to the decrease of omega-6/omega-3 ratio polyunsaturated fatty acids of total phospholipids. NSE modulated the phospholipid membrane composition in both the metastases and the neighbouring conditionally normal lung tissue.     

Chronic low doze ionizing irradiation and polyfactors of adaptation

The analysis of cytogenetic anomaly dynamics in conditions of chronic low doze effect of ionizing irradiation in generations of genetically homogeneous laboratory line of mice CC57W/Mv and genetically heterogeneous populations of vole's species, trapped in alienation zone of Chernobyl's NPP in places with different levels of radio nuclide contamination in 1994-2001 was carried out. The data about the physiological adaptation to ionizing irradiation effects under age of linear mice and non-linearity of frequency changes of separate cytogenetic anomalies in generations as laboratory line of mice, and genetically heterogeneous populations of vole's species were obtained. Was supposed, that the number of non-linear effects of the ionizing irradiation in low doze range could be stipulated by comparability between the intensity of it's damage effects and the activation by it the polyfactor systems of anti oxidant ones, reparation events, and also elimination of defective cells.     

Genomic typing of the Echinococcus granulosus isolates from the areas of Southern Urals

Nine larvocysts of Echinococcus granulosus isolated from nine patients and one cyst derived from a naturally infested cattle have been examined. Genomic typing was carried out in order to identify strains of E. granulosus. All DNA samples were shown to have the same genotype, E. granulosus G1.     

Virus-induced dysregulation of programmed immunocompetent cell death: pathology or adaptation?

The review summarizes information from recent literature and results of the authors' own investigations concerning dysbalance of programmed cell death in establishment of a long-term virus persistense. The article discusses molecular mechanisms of apoptosis modulation of immune cellls by persistent viruses.     

Structure and localization of sensilla on antennas of caddisflies (insecta: trichoptera)

The structures of antennal segments and ultrastructures of antennal sensilla were studied in representatives of 28 families of caddisflies from both extant suborders by the methods of light and scanning electron microscopy. Sixteen types of the sensilla have been found to occur on the antenna in Trichoptera; some of them were found for the first time. Morphological characters of the cuticular structures on the antennal surface demonstrate the significant structural differences both in various families and in the lower taxonomy levels. Specialized sensory fields differing structurally from the rest of the flagellomer surface have been found on the antennas in the suborder Phryganeina. A modified classification of sensilla based on the cuticular structures is proposed.     

Identification of genetically modified vegetable sources in food and feed using hydrogel oligonucleotide microchip

A method of multiplex polymerase chain reaction (PCR) followed by the hybridization on a hydrogel oligonucleotide biochip was developed for simultaneous identification of ten different transgenic elements of plant DNA in feed and food products. The biochip contained 22 immobilized probes intended for (i) detection of plant DNA; (ii) plant species determination (soybean, maize, potato, rice); (iii) identification of transgenic elements, including 35S CaMV, 35S FMV, rice actine gene promoters, nos, 35S CaMV, ocs, pea rbcS1 gene terminators, and bar, gus, nptII marker genes. The limit of detection was 0.5% of genetically modified (GM) soybean and maize in analyzed samples. Identification of transgenic DNA in food and feed products using either the developed approach or real-time PCR led to virtually identical results. The assay can be used for selection of GM samples by screening food and feed products for subsequent quantitative determination of the GM component based on the identified transgene.     

Animal in vivo model of arrhythmia for genes target identification for 5-amino-exo-3-azatricyclo[5.2.1.0(2,6)]decan-4-one

The goal of the current work is to study the molecular mechanisms underlay the action of 5- amino-exo-3-azatricyclo[5.2.1.0(2,6)]decan-4-one (P-11) with combined antiarrhythmic, nootropic, anti-inflammatory and anaesthetic activities. The aconitine-induced experimental rat model of cardiac arrhythmia has been used in our study. Aconitine was administered once intravenously in a dose 50 microg/kg whereas experimental animal group received P-11 in a dose 0.3 mg/kg (the compound was injected intravenously 2 min before acute aconitine treatment). Expression macroarray (Atlas Rat cDNA Expression Array, #7738-1; BD Biosciences) was used to identify the target genes for P-11 compound. Comparative analysis of changes in the status of expression of genes in the heart of rats induced by P-11 against the simulated in vivo arrhythmia identified 16 genes that reproducibly alter the level of expression.These genes encode the extracellular matrix proteins (glypican 1, Gpc1; tissue inhibitor of metalloproteinase 2, 3, Timp2, Timp 3); intracellular signaling molecules (rho GTPase activating protein 7, Dlc1; protein tyrosine phosphatase 4a1, Ptp4a1; phosphodiesterase 4D, PDE4D; PI3-kinase regulatory subunit alpha, PIK3R1; guanine nucleotide binding protein alpha 12, Gna12) and protein of intermediate junctions (junction plakoglobin, Jup), proteins involved in glycolysis (phosphofructokinase I, Pfk1) and hemostasis (tissue plasminogen activator, Plat), plasma membrane transporters (Solute carrier family 16, member 1, Slc16a1; ATPase, Na+/K+ transporting, Atp1a), and ets. (c-fos protooncogene, c-fos; telomerase protein component 1, tlp; Annexin 1, anxa 1). Thus, the data about the selective effect of P-11 on genes whose products are involved in the aritmogenesys mechanisms, allow us to consider this compound as a promising means of pathogenetically oriented pharmacotherapy of cardiac arrhythmias.     

Microarray analytic system for multiplex analysis by real-time polymerase chain reaction with reagents immobilized in microreactors

A microarray analytic system that uses a silicon chip with immobilized in microreactor test-system for multiplex analysis of DNA by real-time polymerase chain reaction (RT-PCR) was developed and optimized. We suggested the method of immobilization of PCR-components of a test-system, chose the stabilizer, and conducted the optimization of the composition of reaction mixture to achieve permanent stability of a microarray. We conducted optimization of preparation of samples using magnetic sorbent and indicated that, with 2.6 x 10(4) copies/ml, 60 min are necessary to obtain positive identification including time for preparation of model probes. The abilities of the created system were demonstrated on the example of microarray analysis of samples with different content of DNA, low absolute limits of identification (20 DNA copies in microreactor), and high reproducibility of the analysis.     

嗜热链球菌胞外多糖EPS333对酸乳品质的影响

本研究以酸乳为基质,通过测试酸乳酸化速率曲线、脱水收缩敏感性、持水力、水分迁移、质构及流变特性等参数,研究了嗜热链球菌胞外多糖与市场常规应用多糖对酸乳品质的影响。研究结果表明:嗜热链球菌胞外多糖EPS333在酸乳凝胶过程中较其他几种多糖能明显减缓酸化趋势,后熟及储藏阶段减少"自由水"含量,降低流动性,改善其流变特性。