Biodistribution of Viscumin after Subcutaneous Injection to Mice and In Vitro Modeling of Endoplasmic Reticulum Stress.

Viscumin (mistletoe lectin I, MLI) in concentrations of 10-11-10-7 M causes endoplasmic reticulum stress and triggers unfolded protein response, a modulator of antitumor immunity, in target cells.

Transcriptomic changes in human umbilical cord blood endothelial cells under simulated microgravity.

Microarray analysis of cultured endothelial cells was performed 24 h after simulated microgravity. A significant change in the expression of 177 genes that can be classified into several functional clusters was detected. Among the genes with overexpression, clusters of cell response to external stimuli and regulation of cell motility and proliferation can be reliably distinguished. Among down-regulated genes, clusters of transcription factors with the "zinc fingers" domain and factors involved in the regulation of morphogenesis and angiogenesis were distinguished. The overlapping of signaling pathways involved in mechanotransduction and inflammatory changes is discussed.

mRNA expression profile of mouse oligodendrocytes in inflammatory conditions.

In this study, we performed transcriptome profiling of oligodendrocyte culture of mice treated with the remyelinating therapeutic agent benztropine in the presence and absence of interferon gamma (IFNγ). The results of this work are important for understanding the expression profile of oligodendrocytes under conditions of systemic inflammation in the central nervous system in multiple sclerosis as well as the mechanisms of cellular response to benztropine in light of its possible use for the treatment of multiple sclerosis.

Comparison of the results of PCR analysis of gene expression in breast cancer tissue specimens stabilized in formalin and RNAlater.

We compared the results of real-time PCR analysis of gene expression in paired specimens breast cancer tissue fixed in RNAlater (Qiagen) stabilization reagent (FF samples) and in formalin (FFPE samples). A clear-cut linear relationship (correlation coefficient 0.76 ± 0.07) was detected between the gene expression logarithm values in FF and FFPE samples. This fact suggests that collections of paraffin blocks with formalin-fixed tissue specimens from patients with a many-year disease history can be effectively used in modern studies.

c-FOS suppresses ovarian cancer progression by changing adhesion.

BACKGROUND: C-Fos was initially described as oncogene, but was associated with favourable prognosis in ovarian cancer (OvCa) patients. The molecular and functional aspects underlying this effect are still unknown. METHODS: Using stable transfectants of SKOV3 and OVCAR8 cells, proliferation, migration, invasion and apoptotic potential of c-FOS-overexpressing clones and controls were compared. Adherence to components of the extracellular matrix was analysed in static assays, and adhesion to E-selectin, endothelial and mesothelial cells in dynamic flow assays. The effect of c-FOS in vivo was studied after intraperitoneal injection of SKOV3 clones into SCID mice, and changes in gene expression were determined by microarray analysis. RESULTS: Tumour growth after injection into SCID mice was strongly delayed by c-FOS overexpression, with reduction of lung metastases and circulating tumour cells. In vitro, c-FOS had only weak influence on proliferation and migration, but was strongly pro-apoptotic. Adhesion to components of the extracellular matrix (collagen I, IV) and to E-selectin, endothelial and mesothelial cells was significantly reduced in c-FOS-overexpressing OvCa cells. This corresponds to deregulation of adhesion proteins and glycosylation enzymes in microarray analysis. CONCLUSION: In addition to its known pro-apoptotic effect, c-FOS might influence OvCa progression by changing the adhesion of OvCa cells to peritoneal surfaces.

Expression profile of inflammatory breast cancer.

Inflammatory breast cancer is characterized by high malignancy, early and rapid lymphogenic and hematogenic metastasizing, and high mortality. The diagnosis of this form of cancer is based fully on the clinical criteria. Whole transcriptome analysis of tumor tissue specimens from patients with inflammatory breast cancer detected 137 differentially expressed mRNA (17 genes with low expression and 120 with high expression). Genes involved in the organization of inflammatory process, chemotaxis, and transcription regulation were active in the process of pathogenesis of inflammatory breast cancer.

[Genetic and biochemical mechanisms of ontogenetic and phylogenetic adaptation]. / Genetiko-biokhimichskie mekhanizmy ontogeneticheskoi i filogeneticheskoi adaptatsii.

Results of the scientific research of genetic mechanisms of ontogenetic and phylogenetic adaptation in the department of genetics and molecular biology of the Odessa national university (ONU) are summarized. The new concept of genetic adaptation and adaptive heterosis is proposed. The evidence are presented that not only selection of selectively valuable alleles of the structural genes of enzymes, but also of gene-modifiers of the gene products are involved in the adaptation of D. melanogaster populations to environmental conditions. As a result, the sets of coadaptation alleles--adaptation complexes of genes (ACG), which are specific to the selective factors, are formed in persons of the population.

[The ADH gene-enzyme system and the fitness of Drosophila melanogaster mutants]. / Gen-énzimnaia sistema ADH i prosposoblennost' mutantov Drosophila melanogaster.

Physico-chemical properties of ADH and some fitness parameters were examined in two mutant (cn and vg) and two wild-type (C-S and D) strains of Drosophila melanogaster. It was shown that, under the experimental conditions, longevity, fecundity and heat resistance did not depend on the activity and the electrophoretic mobility of enzymes. The Adh gene-enzyme system of the mutants was analyzed in relation to the saturation of their genotypes with genes of wild-type flies having different allelic control of the enzyme. ADH activity was shown to be positively correlated with the frequency of F allele of the structural gene (r = 0.84), whereas thermostability of the enzyme was not associated with electrophoretic mobility. Low thermostability of ADH in vg mutants, which was correlated with low heat resistance (r = 0.94), is assumed to be controlled by the thermostable allele Adh Fs.

[Genotypic basis of low viability in vestigial mutants of Drosophila melanogaster]. / Genotipicheskie osnovy nizkoi zhiznesposobnosti mutantov vestigial Drosophila melanogaster.

The role of a marker mutation and other genes in a decrease in viability was studied in the Drosophila melanogaster vg line. In flies of the C-S line, chromosome 2 was substituted by the homologous chromosome of the vg flies. In addition, the flies of the mutant phenotype with mutant genes partially or completely substituted by the wild-type C-S genes were obtained in saturating crosses C-S x vg. In the reciprocal variant of chromosome 2 substitution, the flies of the C-S phenotype with chromosomes 1, 3, and 4 from the vg line were obtained. Chromosome 2 of the vg line, introduced into C-S fly karyotype, proved to substantially reduce the heat resistance and life span of flies. In the case of reciprocal replacement (C-S line chromosome 2 substituted for the homologous chromosome of vg flies), a significant increase in viability was observed, which, however, never reached the level characteristic of the C-S line. As the vg genotype became saturated with C-S genes, the heat resistance and life span of flies increased substantially. However, even the complete saturation of mutant chromosomes with wild-type genes never resulted in the equal viability of vg and C-S flies. These data suggest that the low viability of the vg mutant is largely accounted for by the gene composition of the second chromosome and, primarily, by the presence of the vg gene. Nevertheless, there is evidence that, along with the pleiotropic effect of the marker mutation, other genes not linked to chromosome 2 are responsible for the studied physiological properties of the vg flies.

[The gene-enzyme system of alcohol dehydrogenase during genotype changes in Drosophila melanogaster]. / Gen-énzimnaia sistema alkogol'degidrogenazy pri izmeneniiakh genotipa u Drosophila melanogaster.

It is established, that certain variants of replacement of chromosomes performed on wild (C-S, D) and mutant (cn, vg) lines as well as continuous backcrosses, leads to the change of heat resistance and activity of F- and S-allozymes of ADH in tissues of synthesized forms of drosophila. But nevertheless, the electrophoretic mobility of allozymes does not change. It is assumed that post-translated modifications of ADH play the important part in processes of phylogenetic adaptation of Drosophila melanogaster.

[Gene-enzyme alcohol dehydrogenase system and adaptability in Drosophila melanogaster]. / Gen-enzymna sistema alkogol'degidrogenazy i adaptyvna zdatnist' Drosophila melanogaster.

As follows from the experiments on the genotypic related populations of Drosophila melanogaster with different frequency of ADH allozymes selection for postponement of ageing and resistance to hypothermia lead to saturation of population with S-allozyme ADH, and genotypic adaptation to ethanol-to increasing of frequency of F-allozyme. It is supposed that genotypic adaptation is realized by selection of specimens with the most favourable alleles of genes. At the same time ontogenetic adaptation is accompanied by biochemical modification of existing allozymes.

[Alcohol dehydrogenase polymorphism and the genotypic adaptation of D. melanogaster to the action of selective factors]. / Polimorfizm alkogol'degidrogenazy i genotipicheskaia adaptatsiia D. melanogaster k deistviiu selektivnykh faktorov.

Analysis of experimental populations of Drosophila has shown different selective value of S- and F-alleles of Adh locus under selection for delayed aging, resistance to elevated temperature and ethanol. Selection for delayed aging and resistance to hyperthermia was accompanied by an increase of Adhs frequency in populations and genotypic adaptation to ethanol- by an increase of AdhF frequency.

[The Drosophila melanogaster Adh locus upon selection for delayed aging]. / Lokus Adh Drosophila melanogaster v usloviiakh otbora na zaderzhku stareniia.

Dynamics of the Adh activity and frequencies of alleles AdhF and AdhS were analyzed under selection for delayed senescence. The experiments were performed on Drosophila melanogaster. Lines AdhS cn and AdhF vg and experimental populations cn and vg, selected for an increased duration of reproductive period (late oviposition) were used. Analysis of fertility, longevity, viability and resistance to starvation showed that selection for late oviposition resulted in delayed senescence of flies of the experimental populations. Genetic structure of population vg changed considerably with regard to the Adh locus. This was confirmed by parameters of activity, thermostability and electrophoretic mobility of the enzyme isolated from flies after 30 generations of selection. Analysis of frequencies of the Adh alleles showed that in both selected populations, which initially had different genetic composition, accumulated allele AdhS, which encodes the isozyme that is less active but more resistant to inactivation. Genetic mechanism of delayed senescence in Drosophila is assumed to involve selection at vitally important enzyme loci, including Adh.

[Gene-enzyme system of alcohol dehydrogenase and adaptation of Drosophila to increased temperature]. / Gen-énzimnaia sistema alkogol'degidrogenazy i adaptatsiia drosofily k povyshennoi temperature.

Properties and allelic ADH (alcohol dehydrogenase) control in Drosophila melanogaster were studied upon flies' cultivation under conditions of hypotherm of different intensity and duration. Lines homozygotic for F allele (vg) and S allele (cn) of the Adh gene as well as genetically enriched experimental cn' and vg' populations containing a small number of AdhF/AdhS heterozygotes at the initial stage were used. It was found out that physiological adaptation of the species to momentary influence of elevated temperature is accompanied by modification of physical properties of ADH-F according to ADH-S. Constant influence (during the life span of 25 generations) of elevated temperature on the population changes its genetic structure, due to selective advantages of the S allele of Adh under these conditions.

[Expressivity of the gene-enzymatic system and viability indicators in the ontogenesis of Drosophila inbred lines and hybrids]. / Ekspressivnost' gen-énzimnykh sistem i pokazateli zhiznesposobnosti v ontogeneze inbrednykh linii i gibridov drozofily.

The activity, physico-chemical properties and multiple molecular forms of enzymes (alcohol dehydrogenase, superoxide dismutase, nonspecific alpha- and beta-esterases, hydroxide peptidohydrolase) were studied in ontogenesis of Drosophila inbred lines and their hybrids under conditions of high temperature (37-41 degrees C) and the presence in food of 10% ethanol. It was established that resistance of individuals to the effect of high temperature and alcohol, including manifestation of adaptive heterosis in hybrids not always depends on the level of the activity of enzymes analysed and is rather determined by allelic state of the appropriate structural genes. So, in conditions of the alcohol stress the individuals containing highly active F form of alcohol dehydrogenase have selective advantage and flie with hybrid F/S enzyme of higher activity and heat stability are more stable to the effect of high temperature. It is supposed that the complexes of adaptation genes (CGA) are formed in individuals of populations in response to the regular action of unfavourable environmental factors. These complexes condition optimal allelic control and most efficient regulation of enzyme activity in environment. Genotypic adaptation of individuals as well as occurrence of adaptive heterosis in hybrids seem likely to be connected with formation of CGA.

[Alcohol dehydrogenase and adaptation to ethanol in Drosophila]. / Alkogol'degidrogenaza i adaptatsiia k étanolu u drozofily.

The subject of this research is activity and allozyme spectra of alcohol dehydrogenase (ADH), and survival of mutant strains of Drosophila kept in standard nutrient medium with added ethanol. In all experiments the ADH of flies revealed greater affinity to isopropanol than ethanol. The mutant strains considerably differed from one another and from the wild type of flies in the level of enzyme activity, which may be connected with genotypic properties in the mutants studied. The ADH variability in mutant strains seems to be caused by different alleles of the structural ADH gene, which was established as a result of investigation of activity, electrophoretic mobility and thermostability of corresponding allozymes. As follows from experiments on the genotypical structure of populations in the conditions of fly selection in the medium containing ethanol (10%), the adaptation of flies to exogenous ethanol takes place via mechanisms of allele control of the ADH activity. Phenotypical manifestation of the ADH locus and its effect on the resistance of Drosophila to alcohol are supposed to depend on complex gene interactions determined by the genotype as a whole.

[Uptake of 14C-nicotinic acid by membrane structures of rat tissue in alloxan diabetes with insulin administration]. / Pogloshchenie [14C]nikotinovoi kisloty membrannyme strukturami tkanei krys s alloksanovym diabetom pri vvedenii insulina.

The uptake of nicotinic acid by erythrocytes, hepatocytes and inverted small intestine sacs of rats with alloxane diabetes was studied as affected by insulin. It is established that a disturbance of the pancreas endocrine function caused by the administration of alloxane to rats inhibits the accumulation of [14C]nicotinic acid in all the studied structures. Insulin administered to rats with alloxane diabetes stimulates the acid uptake by erythrocytes and hepatocytes, but has no effect on accumulation of the vitamin in tissues and serous fluid of inverted empty intestine sacs. An assumption is advanced that under alloxane diabetes the membrane transport of nicotinic acid is disturbed and it may be partially normalized by the insulin administration. The efficiency of the hormone influence on the vitamin uptake by the structures under study changes essentially against a background of the actinomycin D and cycloheximide action.

[Possible role of lysosomal proteinases in the biological effects of G-forces]. / O vozmozhnoi roli lizosomal'nykh proteinaz v biologicheskikh éffektakh peregruzki.

Experiments on rats demonstrated that acceleration of 6 g for 10 min elevated significantly the activity of acid proteinases and the rate of their release from lysosomes into the incubation mixture. Hydrocortisone (1 mg/100 g body weight) also increased proteinase activity; however, this effect was transient and less expressed. Upon a combined effect of hydrocortisone and acceleration the effect of the latter on the activity of lysosomal enzymes weakened appreciably.