Accumulation of colicin M protein and its biological activity in transgenic lettuce and mizuna plants.

Food-borne illnesses caused by pathogenic strains, especially enterohaemorrhagic (EHEC), are a serious public health problem, as debilitating disease and even death from such food poisonings have been repeatedly reported. Colicin M (ColM), a non-antibiotic antimicrobial protein produced by some strains of , has shown promising activity in controlling multiple enteropathogenic strains of and related pathogens. As contaminated green leafy vegetables are a frequent source of pathogenic infections, we genetically modified (GM) two edible crops, lettuce ( L.

[Changes of the genome-wide DNA methylation level in various regions of the rat brain with incomplete cerebral ischemia].

Objective: To quantify genome-wide DNA methylation in the olfactory bulbs, fronto-parietal and occipital regions, and cerebellum in normal male Wistar rats and in modeling incomplete cerebral ischemia caused by permanent bilateral occlusion of the common carotid arteries.

Material And Methods: The study was performed on 23 male Wistar rats divided into groups: «Sham operation» and «Cerebral ischemia». The level of genome-wide methylation of CCGG sites was determined by methyl-sensitive restriction using MspI/HpaII endonucleases followed by densitometric analysis of electrophoregrams by ImageJ software.

[DNA methylation in experimental ischemic brain injury].

The enrichment of angioneurology with fundamental advances leads to the understanding of new important facets in the pathogenesis of cerebral ischemia. The knowledge of epigenetic mechanisms in the development of stroke, in particular, DNA methylation, which makes a significant contribution to the development and formation of cerebral damage, is becoming more and more relevant. This review reflects an analysis of animal studies proving the relationship of DNA methylation with cerebral ischemia.

Rhizoextraction Potential of Convolvulus tricolor Hairy Roots for Cr, Ni, and Pb Removal from Aqueous Solutions.

This study evaluated the potential of dwarf morning-glory Convolvulus tricolor (Convolvulaceae) plants and their hairy roots induced by Agrobacterium rhizogenes for rhizoextraction of heavy metals ions from the liquid medium under aseptic growth conditions. Both the young C. tricolor plants and the generated hairy root culture efficiently removed Cr, Ni, and Pb ions from the liquid cultivation medium.

[CHANGE IN BCL-2 PROTEIN EXPRESSION IN NEURONS OF THE HIPPOCAMPAL AREAS AFTER THE APPLICATION OF ISCHEMIC CEREBRAL POSTCONDITIONING].

Bcl-2 protein expression was studied in hippocampal CA1, CA2, CA3 and CA4 pyramidal neurons in Mongolian gerbils (Meriones unguiculatus), of the in in the early (Day 2) and late (Day 7) reperfusion period after a 7-minute forebrain ischemia and following ischemic postconditioning (IPostC), as well as in sham-operated animals (n=60). In the latter, the highest level, of Bcl-2-expression was found in CA4 neurons, while the lowest--in-CA1 neurons (P<0.01).

[Morpho-functional changes of the pyramidal neurons in various hippocampal areas after the ischemic postconditioning].

The aim of this study was to determine the effect of ischemic postconditioning (IP) on the viability of neurons in various hippocampal areas as well as on cytoplasmic activity of succinatedehydrogenase (SDH) in these cells in 30 male Mongolian gerbils (Meriones unguiculatus). Ischemic brain injury was induced by bilateral common carotid artery occlusion for 7 min. IP protocol comprised 3 cycles of 15 s of reperfusion/15 s of ischemia.

Lox-dependent gene expression in transgenic plants obtained via Agrobacterium-mediated transformation.

Lox sites of the Cre/lox recombination system from bacteriophage P1 were analyzed for their ability to affect on transgene expression when inserted upstream from a gene coding sequence adjacent to the right border (RB) of T-DNA. Wild and mutated types of lox sites were tested for their effect upon bar gene expression in plants obtained via Agrobacterium-mediated and biolistic transformation methods. Lox-mediated expression of bar gene, recognized by resistance of transgenic plants to PPT, occurred only in plants obtained via Agrobacterium-mediated transformation.

Activity of succinate dehydrogenase in the neocortex and hippocampus of Mongolian gerbils with ischemic and reperfusion brain injury.

We analyzed changes in activity of SDH, one of the most important enzymes of the Krebs cycle, in the cytoplasm of hippocampal and cortical neurons of Mongolian gerbils (Meriones unguiculatus) at the early and delayed reperfusion period after global brain ischemia. The data indicate that SDH activity in pyramidal neurons of various hippocampal areas and in neurons of II, III and V layers of cerebral cortex after 7-min forebrain ischemia depends on both the localization of these neurons and duration of the postischemic reperfusion. SDH activity in neurons significantly increased on days 2 and 7 after reperfusion.

[Morpho-functional changes of hippocampal CA1 area in Mongolian gerbils after ischemic postconditioning].

The aim of this study was to determine the effect of ischemic postconditioning on hippocampal CA1 neuronal survival and cytoplasmic activity of lactate dehydrogenase (LDH) in the gerbil model of cerebral ischemia-reperfusion injury. Ischemia was induced by bilateral common carotid artery occlusion (for 7 min) in male Mongolian gerbils (Meriones unguiculatus). Ischemic postconditioning protocol comprised 3 cycles of 15 s reperfusion/15 s ischemia.

The infarct-limiting effect of cerebral ischaemic postconditioning in rats depends on the middle cerebral artery branching pattern.

Cerebral ischaemic postconditioning (PostCon) is a recently discovered endogenous neuroprotective phenomenon that occurs after several brief bouts of reperfusion/ischaemia instituted immediately after prolonged cerebral ischaemia. Data on the extent of PostCon-mediated infarct size limitation in models of focal cerebral ischaemia-reperfusion are controversial. In this study, we investigated the infarct-limiting effect of PostCon in the rat model of focal cerebral ischaemia-reperfusion.

[The impact of early and late ischemic preconditioning on brain damage and degree of neurological deficiency in rats].

The present study was aimed to investigate neuroprotective effects of early and late ischemic preconditioning in the acute phase of ischemic brain damage in rats. It was found that a single five-minute ischemic episode of early ischemic preconditioning did not lead to significant neuroprotective effects in comparison with control group, while three five-minute ischemic episode early ischemic preconditioning accompanied by a significant increase in neurological deficit and growing damage rate in CA1 hippocampus neurons. In contrast, later ischemic preconditioning in form of single five-minute episode 24 hours before ischemia modeling, provided a significant neuroprotective effect, manifesting reduced neurological deficit and maintaining viability of CA1 hippocampus neurons.

[Molecular mechanisms of development of cerebral tolerance to ischemia (Review. Part 2)].

In the 2nd part the authors describe in details the main aspects of protective effect of preconditioning of the brain: inhibition of programmed cell death, weakening of phenomenon of excitotoxicity, activation of endogenous antioxidant systems, anti-inflammatory effects, modulation of glial cell function, changes in regional blood flow and vascular reactivity. In addition, data analysis on the impact of preconditioning on brain neurogenesis, the state of the blood-brain barrier, ion homeostasis and metabolism of neurons is presented. Review emphasizes the role of microRNAs in mechanisms of ischemic tolerance of brain.

[Molecular mechanisms of development of cerebral tolerance to ischemia. Part 1].

In the first part of this review molecular mechanisms of ischemic tolerance emerging as a result ofpreconditioning of the brain are discussed. Data on inductors, sensors, transducers and effectors of early and delayed ischemic tolerance are presented.

A new rat model of reversible global cerebral ischemia.

A new rat model of global cerebral ischemia-reperfusion was proposed via reversible occlusion of the major vessels originating from the aortic arch and supplying the brain. This technique can be used for the search and study of exogenous (pharmacological) and endogenous methods of brain protection from ischemia-reperfusion injury.

[Activity of lactate dehydrogenase in the brain cortex and hippocampus of Mongolian gerbils after global ischemia and reperfusion injuries].

Cerebral ischemia results in severe derangements of energy metabolism in the nervous tissue including activation of glycolytic pathway. Activity of cytosolic lactate dehydrogenase (LDH) in the specific brain structures remains unclear. The recent study was aimed at investigation into the LDH activity in the cytoplasm of both hippocampal and cortical neurons in Mongolian gerbils (Meriones unguiculatus) at different durations of reperfusion after global ischemia.

Expression of genes for thioredoxin 1 and thioredoxin 2 in multidrug resistance ovarian carcinoma cells SKVLB.

The expression of genes for thioredoxin isoforms Trx1 and Trx2 was studied in sensitive SKOV-3 and resistant SKVLB human ovarian carcinoma cells. The development of doxorubicin resistance was accompanied by a significant increase in the expression of TRX1 gene and less pronounced increase in TRX2 gene expression.

Expression of genes for redox-dependent glutathione S-transferase isoforms GSTP1-1 and GSTA4-4 in tumor cell during the development doxorubicin resistance.

Expression of genes for redox-dependent glutathione S-transferase isoforms GSTP1-1 and GSTA4-4 in tumor cells K562, MCF-7, and SKOV-3 was studied during the development of resistance to doxorubicin. It was found that the development of resistance was accompanied by predominant increase in the expression of hGSTP1 gene in MCF-7 cells, and hGSTA4 gene in resistant K562/DOX and SKVLB cells.

[A comparative study of induction regulation in cytochromes family 1 P450 in cell cultures at different stages of tumor transformation].

Lipophilic xenobiotics, including some carcinogenic agents and cytostatics, are metabolized by cytochrome P450 isoforms (CYP). In tumours expression of CYP genes and their inducibility are lower than in a homologous normal tissue. This phenomenon determines the known higher cytostatic stability of tumour cells.

[Changes in the free-radical state and level of free iron during the formation of drug resistance in tumor cells].

The development of resistance of K562 human erythroleukemia cells to doxorubicin, a widely used antitumor antibiotic with the prooxidant action, leads to changes in the free-radical state of cells. It has been found that the formation of superoxide anion in resistant cells decreases. The introduction of doxorubicin to the culture medium induced a considerably lesser increase in the formation of O2*- in resistant cells compared to sensitive cells.

[Effect of lox-sites of the Cre lox recombination system on promoterless bar gene expression in transgenic plants].

We demonstrate that localization of lox site between the right border of T-DNA and promoterless bar gene (RB-lox-bar-) led to its highly efficient expression in transgenic plants of Nicotiana tabacum and N. africana. Plasmid vectors used in gene integration experiments contained neomycin phosphotransferase II (npt II) gene under nos promoter as well.

[Genetic transformation of Nicotiana africana Merxm. with plasmids containing lox recombination].

An efficient genetic transformation method for african tobacco Nicotiana africana Merxm. has been established. African tobacco is a valuable source for cytoplasmic male sterility (CMS) and nuclear encoded resistance to potato virus Y (PVY).

[In vitro regeneration of Nicotiana africana plants from explants of different type and mesophyll protoplasts].

Methods of in vitro cultivation of an African endemic species Nicotiana africana (Solanaceae) possessing some valuable traits have been elaborated. Influence of different concentrations of auxin (alpha-naphtaleneacetic acid) and cytokinins (6-benzylaminopurine, zeatin, thidiazuron) on morphogenesis and plant regeneration has been analysed using leaves and internodes as explants. The optimum method of regeneration of N.

Cytochrome P-450 family 1 in rat embryo cell culture immortalized by Rausher leukemia virus.

We studied comparative expression and activity of cytochrome P450 family 1 (CYP1) isoforms in rat embryo cells, both primary and immortalized by Rausher leukemia virus (RLV). In RLV-infected embryonal cells compared with the initial ones the expression levels of CYP1A1 and 1B1 mRNAs and benzo[a]pyrene (BP) hydroxylase activity were higher, regardless of their treatment with the CYP1 inducer 2,3,7,8-tetrachlorodibenzo-p-dioxin. The sensitivity to BP and 7,12-dimethylbenzo[a]anthracene was higher in the cells immortalized with RLV.

[Morphological and cytogenetic disorders in rats following chronic exposure to elevated radiation].

Object of the investigation were effects of elevated radiation background on specific body systems in two generations of rats that had been chronically exposed to radiation in the most badly polluted areas of the Kaluga district and eaten local food. Findings included some compensatory deviations in the white blood count, decrease in karyocytes and increase in chromosomal aberration rate in nucleus-containing cells from the femoral bone marrow in younger animals of several groups, morphologic and histochemical shifts in mucous membrane of the esophagus and the stomach, and morphofunctional changes in neurons and glia of the cerebral cortex. These changes are compensatory-adaptive by nature and indicative of destabilization of homeostasis in experimental animals.

[Role of the antioxidant system and redox-dependent regulation of transcription factors bcl-2 and p53 in forming resistance of human K562 erythroleukemia cells to doxorubicin].

Prooxidant effect of chemotherapeutic agents is of significant interest in connection with activation of oxidative stress in cancer cells. Role of development of adaptive antioxidant response to the rise of resistance to cytotoxical effect of doxorubicin (DOX) has been studied in human erythroleukemia K562 cells. Growth of resistance to DOX caused enhancement of antioxidant enzymes (Cu, Zn-SOD, Mn-SOD, catalase) elevation of Mn-SOD activity being predominant.