Detection of Taxuyunnanin C in Suspension Cell Culture of Taxuscanadensis.

This is the first study to isolate the taxoid taxuyunnanin C (group of 14-hydroxylated taxoids) from the biomass of suspension cell culture of the Canadian yew (Taxus canadensis). According to available data, this is the first report of the presence of nonpolar (polyacylated) forms of 14-hydroxylated taxoids, including taxuyunnanin C, in T. canadensis.

Occurrence of 14-hydroxylated taxoids in the plant in vitro cell cultures of different yew species (Taxus spp.).

This is the first study to show that the formation of 14ß-hydroxylated derivatives of taxa-4(20),11-diene is a specific feature of in vitro cultured dedifferentiated yew cells that distinguishes them from intact plant cells. This may be due to a lower toxicity of the 14-OH taxoids for proliferating plant cells compared to the 13-OH derivatives.

[Formation of protodioscin and deltoside isomers in suspension cultures of Nepal yam (Dioscorea deltoidea Wall.) cells].

Changes in the content of the furostanol glycosides protodioscin and deltoside, particularly that of the (25S)-isomers of the glycosides, during suspension cultivation of different lines of Nepal yam (Dioscorea deltoidea Wall.) cells of the strain IFR-DM-0.5 has been investigated. The composition of furostanol glycosides has been characterized, and the dynamics of the accumulation of individual glycosides during lengthy subcultivation of cells maintained in flasks or in a barbotage bioreactor has been analyzed. A positive correlation between the growth and accumulation of substances that belonged to the class of furostanol glycosides has been demonstrated for cultured dioscorea cells, whereas the content of some of the individual glycosides varied considerably between the lines of the strain, cultures maintained under different conditions, and even between cells in different phases of the growth cycle. The increased content of (25R)-forms of the glycosides (protodioscin and deltoside) was correlated with a decrease in the cellular growth rate, whereas an increase in culture growth intensity occurred concomitantly to an increase of the amount of (25S)-isomers. This may be indicative of the specific stimulatory effect of (25S)-glycosides, but not the (25R)-forms, on cell proliferation in vitro. Thus, the concentration of (25S)-forms may increase due to the autoselection of cells capable of intensive division during prolonged cultivation.

[Use of the tactics of multistage surgical treatment on the model of combined radiation-and-mechanic injury].

Use of the tacticts of multistage surgical treatment on the model of combined radiation-and-mechanic injury. The article provides information on the use of the tactics of multistage surgical treatment (MST), which allows increasing survival value in patients with polytrauma. There are no descriptions of possibility of the use of this tactics in case of combined radiation-and-mechanic injuries in available literature. During an experimental research underwent on 25 sheep it was found that in case of MST implementation there is a significant decrease of intraoperative and postoperative lethal outcome in comparison with traditional tactics. In animals, underwent MST, were defined more laudable course of radiation syndrome. Analysis of laboratory peripheral blood indexes allowed making conclusion, that conducting final surgical intervention (3'd stage of MST) during the first 24 hours after abbreviated surgical intervention allows finishing surgical treatment and intensive care before the beginning of acute radiation syndrome and doesn't counter classic recommendations concerning the treatment of combined radiation-and-mechanic injuries.

PHYSIOLOGICAL, CYTOLOGICAL AND BIOCHEMICAL STABILITY OF Medicago sativa L. CELL CULTURE AFTER 27 YEARS OF CRYOGENIC STORAGE.

BACKGROUND: The efficiency of long-term cryogenic storage to prevent somaclonal variations in plant cell cultures and retain their major cytogenetic and biochemical traits remains under debate. In particular, it is not clear how stress conditions associated with cryopreservation, such as low temperature, dehydration and toxic action of some cryoprotectants (DMSO in particular), affect post-storage regrowth and genetic integrity of cell samples. OBJECTIVE: We assessed growth, cytogenetic and biochemical characteristics of the peroxidase-producing strain of Medicago sativa L. cell culture recovered after 27 years of cryogenic storage as compared to the same culture before cryopreservation. MATERIALS AND METHODS: In 1984, M. sativa L. cell culture was cryopreserved using programmed freezing and 7% DMSO as a cryoprotectant. In 2011, after rewarming in a water bath at 40 degree C for 90 s, cell culture was recovered and proliferated. Viability, growth profile, mitotic index, ploidy level, peroxidase activity and cell response to hypothermia and osmotic stress were compared between the recovered and the initial cell cultures using the records available from 1984. RESULTS: Viability of alfalfa cell culture after rewarming was below 20% but it increased to 80% by the 27th subculture cycle. Recovered culture showed higher mitotic activity and increased number of haploid and diploid cells compared to the initial cell line. Both peroxidase activity and response to abiotic stress in the recovered cell culture were similar to that of the initial culture. CONCLUSION: Cryopreservation by programmed freezing was effective at retaining the main characteristics of M. sativa undifferentiated cell culture after 27 years of storage. According to available data, this is longest period of successful cryopreservation of plant cell cultures reported so far. After storage, there was no evidence that DMSO had any detrimental effect on cell viability, growth or cytogenetics.

[Ginsenosides affect the system of Toll-like receptors in the brain of rats under conditions of long-term alcohol withdrawal]. / Ginzenozidy vliiaiut na sistemu Toll-podobnykh retseptorov v strukturakh golovnogo mozga krys v usloviiakh otmeny dlitel'noi alkogolizatsii.

Long-term alcohol consumption causes the development of neuroinflammation in various brain structures. One of the mechanisms involved in this process is the increased activity of TLR-signaling intracellular pathways. Studies confirm the ability of ginseng extract or its individual ginsenosides to reduce the increased activity of TLR-signaling pathways. The aim of our study was to study the effect of the amount of ginsenosides obtained from the extract of the Panax japonicus cell line on the state of the TLR-signaling system in the nucleus accumbens and hippocampus of the rat brain in a model of long-term alcohol consumption during alcohol withdrawal. The results of the study showed that ginsenosides were able to make changes in the TLR signaling system, which has been altered by long-term alcohol consumption. A significant effect of ginsenosides on the level of TLR3 and TLR4 mRNA in the nucleus accumbens was found, while in the hippocampus, ginsenosides significantly affected the level of TLR7 mRNA. The effect of ginsenosides on the level of mRNA of transcription factors and cytokines involved in TLR-signaling was evaluated. Thus, results of our study confirm that ginsenosides are able to influence the state of TLR-signaling pathways, but this effect is multidirectional in relation to different brain structures. In the future, it seems interesting to evaluate the role of individual ginsenosides in relation to genes of TLR-signaling, as well as the effect of ginsenosides on other brain structures.

[Breathing activity of suspension culture of cells of Polyscias filicifolia Bailey, Stephania glabra (Roxb.) Miers, and Dioscorea deltoidea Wall].

Peculiarities of breathing of cultures of cells producing biologically active compounds (isoprenoids and alkaloids) were investigated in order to optimize productivity of culture growth and biosynthesis. It had been revealed that studied cultures of cells of Dioscorea deltoidea Wall (producer of furistanol glycosides), Stephania glabra (Roxb.) Miers (producer of stepharin alkaloid) and Polyscias filicifolia Bailey (complex of biologically active agents) differ both in joint breathing activity and in ratio between cytochrome and cyanide-resistant breathing, while changes of rate of total oxygen consumption and activity of alternative oxidase during growth were found to be individual for every investigated culture. Maximum rate of oxygen consumption for cells of D. deltoidea and S. glabra was marked in the period preceding active synthesis of secondary metabolites (lag phase for D. deltoidea and exponential phase for S. glabra). The revealed trends can be used for further monitoring and regulation of growth and biosynthesis of secondary metabolites in producing cell cultures during deep cultivation.

[Comparative expression in Escherichia coli of the native and hybrid genes for acyl-lipid delta(9) desaturase].

Expression of the desC gene coding for acyl-lipid delta(9) desaturase of thermophilic cyanobacterium Synechocystis sp. PCC6803 was studied in Escherichia coli cells. In a hybrid gene constructed (desC-licBM3), a sequence of the native acyl-lipid delta(9) desaturase was fused in frame with the reporter gene coding for thermostable lichenase. Lichenase contained in the hybrid protein simplified selection and analysis of the expression of membrane desaturase in the heterologous host. Comparisons of the expression for the native and hybrid genes in bacterial cells showed that lichenase remained active and thermostable in the hybrid protein, while desaturase retains the capability of introducing a double bound in the corresponding position of fatty acids.

[Adaptogenic effects of furostanol glycosides of Dioscorea deltoidea wall on oxidative processes in tomato plants in biotic stress].

The effect of furostanol glycosides of cell culture of Dioscorea deltoidea Wall on oxidative processes in tomato plants subjected to invasion with the gall nematode Meloidogyne incognita Kofoid et White was studied. We showed that furostanol glycosides induce a nonspecific defensive response in plants. Exposure of cell membranes to furostanol glycosides cause rearrangements in fatty acids resulting in the formation of conjugated dienes, which makes molecules thermodynamically more stable under stress conditions. The study of changes in the activity of peroxidases of intact plants and plants affected with the nematode, which were treated with furostanol glycosides, showed that the protective effect of the guaiacol-dependent peroxidase is more long-term than the effect of the benzidine-dependent peroxidase.

Antimicrobial activity of Polyscias filicifolia cell biomass extracts.

Antibacterial activity of extracts of Polyscias filicifolia biomass from bioreactor and callus was determined using the agar disc-diffusion method. The microorganisms Staphylococcus aureus (three strains) showed the highest sensitivity to extracts of P. filicifolia biomass from a bioreactor. The values were comparable with nitrofurantoine used as a standard. Micrococcus flavus, Sreptococcus pyogenes and S. agalatiae were less sensitive. The effect of P. filicifolia callus extract on the above bacteria was less pronounced than that of extracts of biomass from a bioreactor.

[Biosynthesis of mini-antibodies to human ferritin in plant and bacterial producers]. / Biosintez mini-antitel k ferritinu cheloveka v rastitel'nykh i bakterial'nykh produtsentakh.

A recombinant scFv antibody against human spleen ferritin was expressed as a barstar-fused protein in Escherichia coli and in Nicotiana tabacum plants and suspension cell cultures. As demonstrated by immunoblotting with antibarstar antibodies, direction of the recombinant protein to the endomembrane system of plant cells ensured its stability and solubility. Production of the recombinant protein did not differ between parental transgenic plants and their first-generation progeny. Fusion with barstar allowed not only immunochemical detection of the recombinant scFv antibody, but also their purification from the plant material by affinity chromatography with barnase-His6 immobilized on a metal-affinity carrier.

[The complex study of the epidemic situation and of the immunological structure of the population in brucellosis foci]. / Kompleksnoe izuchenie épidemicheskoi situatsii i immunologicheskoi struktury naseleniia v ochagakh brutselleza.

As the result of the complex clinico-laboratory examination of 265 cattle breeders working on 5 farms where cases of brucellosis among agricultural animals had been detected, 62 persons (21.8%) were found to have the positive reaction to brucellosis. A strikingly high proportion (38.3%) of infected persons was found among the workers of a cattle-feeding complex. Out of 62 persons with the positive reaction to brucellosis, 32 had different clinical manifestations of the disease. Treatment for all persons who needed it was organized, the rest were placed under dispensary observation. Regular complex clinico-laboratory examinations of cattle breeders to brucellosis were recommended.

[Epidemiologic characterization of Brucellosis under modern conditions]. / Epidemiologicheskaia kharakteristika brutseleza v sovremennykh usloviiakh.

The spread of brucella infection under modern conditions of cattle breeding were studied with the foci in the Rostov region taken as an example. The main epidemiological features and tendencies of epidemiological importance were established. To find out specific antigen in different environmental objects, the approbation of the enzyme immunoassay test systems was carried out and the possibility of their use at the institutions of the State Sanitary and Epidemiological inspection was shown.