Change of state of endothelial cells occurs under the action of viral infection and bacterial lipopolysaccharide (LPS) that leads to cell dysfunction. Therefore, the aim of the current study was to investigate the effect of LPS from Escherichia coli and influenza A virus on proliferative activity of human endothelial cells (ECV-304) and gene expression of several cytokines and cellular factors: TNFá, TGFâ, IFN-ã, MMP-9, NF-êB, Rho A, eNOS and iNOS. It was found that ECV-304 cells once infected with very low infectious doses of influenza virus acquire the ability to long-term active proliferation (over 8 passages). Addition of LPS E. coli reduced the virus-stimulated cell proliferation. It was shown that influenza virus and LPS can affect on gene expression of cytokine and other cellular factors. When endothelial cells had been infected with influenza A virus in the presence of LPS, there was a significant increase in the expression of several genes and replacement of some genes expression on the expression of other genes. Expression of MMP-9 gene was inhibited in the case of separate exposure to the virus and LPS, but it was significantly increased during the first day under the adding of the virus and LPS together, as well as the activity of the IFN-ã gene; gene of TNFá was active for only 1—3 days whereas genes expression of other factors (TGFâ, eNOS, iNOS, NF-êB and Rho A) increased significantly at the 5th day as in the case of adding only LPS. Thus, the change of physiological state of endothelial cells occurs in the presence of influenza A virus and LPS and it can be caused during different time periods (as well as by varying degrees of virus infection of cells) by different cellular factors and possibly with involvement of different signaling pathways.

Mouse embryonal fibroblasts with knockout of CDKN1A gene encoding p21/Waf1 protein transformed by oncogenes E1A and cHa-ras (mEras-Waf1–/– cell line) have been used to assess the level of DNA repair genes expression — Rad51 and XRCC5 after treatment with HDAC inhibitor sodium butyrate as compared with their control counterparts (mEras-Waf1+/+ cells). mEras-Waf1–/– cells are characterized by the elevated amount of single-stranded DNA breaks and g-H2A.X histone foci associated with these breaks. According to immunofluorescence and immunobloting data, Rad51 and Ku80 proteins are highly expressed in the nuclei of both studied cell lines. The level of Ku80 is higher in cells with CDKN1A gene knockout. When cells were treated with DNA-damaging agent adriamycin, there was an additional accumulation of Rad51 foci in the nuclei. However, sodium butyrate reduced considerably the content of Rad51 and Ku80 proteins both in mEras-Waf1+/+ and mEras-Waf1–/– cells as well as in the cells treated by adriamycin. RT-PCR and immunobloting data show that inhibitory effect of sodium butyrate takes place at the level of Rad51 and XRCC5 gene transcription and the content of Rad51 and Ku80 proteins. The observed suppressive effect of HDACI on DNA repair components explains in part the mechanisms of antiproliferative function of HDAC inhibitors. Surprisingly, sodium butyrate was shown to activate the pluripotent genes transcription in mEras-Waf1+/+ and mEras-Waf1–/– cells, as exemplified by upregulation of Oct-4, Sox-2, Klf4, implying that these pluripotent genes are under negative control at the level of chromatin structure.

We have studied the effect of new ligands of progesterone receptors, including pregna-D'-pentaran 6-methoxyimino-16a,17a-cyclohexanopregn-4-en-3,20-dio-ne (K1047), 17a-acetoxy-3b-butanoyloxy-6-methylpregna-4,6-dien-20-one (buterol), progesterone (P4), and medroxyprogesterone acetate on the viability of HeLa cells and expression of estrogen receptor-alpha (Era) mRNA gene in these cells. K1I047 and buterol exhibited high cytostatic activity, which exceeded the activity of reference compounds on the average by 15% (p < 0.05). Both buterol and K-1047 (at 10(-6)M) effectively suppressed ERa mRNA gene expression in HeLa cell culture by 83.4 - 9 8.6%.

The effect of sulfated polysaccharides (PS) from brown alga Fucus evanescens and their enzymatic transformation and low molecular weight product on the functional activity of the innate immunity cells, i.e. polymorphonuclear leukocytes of human peripheral blood (NF) was comparatively studied. The in vitro NF contact with PS resulted in significant changes in the functional activity of NF, evident from higher density of molecules CD69, CD14, CD11b on the cell membranes with simultaneous lowering of that of CD62L and increased phagocytic and bactericidal activity of NF. The low molecular weight product resulting from fucoidan transformation with fucoidanases showed a higher effect on the level of the molecules CD14, CD11b and CD62L expression vs. the high molecular weight PS.

The functional importance of DNA methylation, which is a special case of epigenetic variation, is meant for regulation of many biological processes, ranged from tissue specific gene expression to remodeling of chromatin structure. Disorders of the DNA methylation can cause changes in the cell's phenotype, providing a significant impact on the development of pathology. Both exogenous and endogenous factors are able to cause disruption of DNA methylation, while epigenetic changes usually precede the emergence of clinical and morphological symptoms of pathological process development, consequently the parameters of DNA methylation can be used as sensitive biomarkers to detect adverse effects on the organism. The purpose of the study was to identify genes of the liver, the methylation profile of which changes under the influence of hepatotoxicants of different nature. The experiment was carried out on 60 male Wistar rats with initial body weight (b.w.) 83.3±1.5 g. Animals were randomly divided into 6 groups - 1 control and 5 test groups, with 10 rats in each group. During the first two weeks of the experiment the rats of the 1-5th test groups were administered to aflatoxin B1 (200 Mg/kg b.w.), cadmium chloride 2,5-hydrate (2 mg/kg), monosodium glutamate (1000 mg/kg), epigallocatechin gallate (EGCG) (1000 mg/kg), paracetamol (150 mg/kg), accordingly. Methylation of the liver genes in rats was determined by using high-performance methods, based on bisulfite sequencing of reduced representation. For each sample from 12 to 30 million pairs of reads were received, genes which demonstrated significant changes in methylation when exposed to toxic factors were identified: aflatoxin B1 caused changes in the methylation of 57 genes; cadmium - 54 genes; monosodium glutamate - 39 genes; EGCG -198 genes; paracetamol - 167 genes. The comparison of genes with altered methylation in the experimental groups revealed that none of the genes repeatedly occurred under the influence of each toxicant out of five, the highest number of repeats accounted 3. As a result of the present analysis 7 genes have been selected: methylation change in Fan1 gene was observed when exposed to cadmium, monosodium glutamate, EGCG; gene Lppr2 - under the influence of aflatoxin B1, EGCG, paracetamol; gene Mlh3 - under the influence of aflatoxin B1, cadmium, paracetamol; Sirt7 gene - under the influence of cadmium, EGCG, paracetamol; gene Fbxo15 - when exposed to cadmium, monosodium glutamate, paracetamol; gene E2f1 - when exposed to cadmium, EGCG, paracetamol; gene Mrps16 - when exposed to cadmium, EGCG, paracetamol. On the basis of the received data the project of the panel of genes-biomarkers of toxic effect, including genes Fan1, Lppr2, Mlh3, Sirt7, Fbxo15, E2f1, Mrps16 has been formed.

The purpose of the study was to determine the effects of rutin (R) and hesperidin (Hes), the main representatives of two most studied subclasses of flavonoids - flavonols and flavanones, on the expression of prototypical Nrf2 and AhR-regulated genes and CYP3A1 gene in rats intoxicated with carbon tetrachloride (CCl4). Investigations were carried out on 5 groups of male Wistar rats with the initial body weight (b.w.) 180-200 g (n=40). Rats of the control group and the 1st experimental group received for 14 days the semisynthetic diet, rats of the 2nd experimental group - the same diet plus R (400 mg/kg b.w.), the animals of the 3rd experimental group received the diet with Hes in the same amount, of the 4th experimental group - diet with R (400 mg/kg b.w.) and Hes (400 mg/kg b.w.). Animals of the experimental groups 24 hours before the end of experiment were injected intraperitoneally CCl4 at a dose of 0.5 ml/kg b.w. in olive oil; rats of the control group were injected equal amount of olive oil. For gene expression assessment the mRNA content of NAD(P)H-quinone oxidoreductase (NQO1), heme oxygenase-1 (Hmox1), Nrf2 (Nrf2), AhR (AhR), CYP1A1, CYP1A2, CYP3A1 and β-actin (Actb) in rat liver was determined by real-time RT-PCR. The results showed that in rats intoxicated with CCl4, enrichment of the diet with R, but not with Hes, led to a significant increase in the expression of genes Hmox1, NQO1 and CYP3A1. Combined intake of R and Hes with the diet led to additivity of their action on the expression of Hmox1 gene and to synergism in the effect on the expression of genes NQO1 and CYP3A1. A moderate increase in the levels of expression of AhR and CYP1A2 genes as compared to their expression in rats treated with CCl4 only, CCl4 and R or CCl4 and Hes has been noted. Thus, for the first time on the model of oxidative stress in rats the data have been obtained showing at the gene expression level a synergism of action of two flavonoids - R and Hes, widely present in the daily human diet.

Peptide KED (Lys-Glu-Asp) has vasoprotective effects and is effective substance in treatment of of atherosclerosis and other cardio-vascular disorders in elderly people. One of the probable mechanisms of biological activity of this peptide is epigenetic genes regulation. These genes can coding proteins, which are markers of endothelium functional activity. The goal of investigation was to study the KED peptide effect on signal molecules expression in normal, atherosclerotic and restenotic endothelium in vitro. It was shown, that KED peptide has normalized endothelin-1 expression, which increased during atherosclerosis and restenosis. KED peptide also restorates cells interactions by connexin expression. Geroprotective effect of KED peptide is realized by increasing of sirtuin1 expression, which has took part in DNA reparation.

to elucidate an influence of nerve growth factor mimetic GK-2 on the expression of neurotrophic factors and the process of neuronal death after ischemia-reperfusion. Materials and methods. Adult white male rats underwent cardiac arrest for 12 minutes, followed by resuscitation. 10 rats were injected GK-2 (Img/kg i/ρ) at 30 minutes and 48 hours after resuscitation. 10 untreated animals received equivalent doses of saline. The control group consisted of sham-operated animals (n = 10). On the 7th postoperative day the total density of hypoxia-sensitive cerebellar Purkinje cells was determined by morphometric analysis. Immunohistochemical study of proteins FGFb, NT4, BDNF was performed by indirect peroxidase-antiperoxidase method using primary polyclonal antibodies. The number of neurons with different expression levels of the neurotrophic factors was determined.

The major problem in prostate cancer treatment is the development of drug resistance and especially important, cross-resistance. The mechanisms of drug resistance, which are divided into ligand-dependent (requiring the presence of androgens in the cell) and independent (not requiring the presence of androgens) are reviewed. The mechanisms are mainly represented with mutations of the androgen receptor and expression of aberrant constitutively active splice variants, as well as up-regulation of genes involved in androgens synthesis.

The effects of exogenous jasmonic acid (JA) on antioxidant enzymes in four-week-old leaves of wild-type Arabidopsis thaliana L. (Columbia-0) and jin1 (jasmonate insensitive 1) mutant plants with defective jasmonate signaling were investigated under normal conditions and under salt stress (200 mM NaCl, 24 h). The wild-type plants responded to JA by an increase in the activities of Cu/Zn superoxide dismutase, catalase, and guaiacol peroxidase, while there was no change in the case of the mutant plants. In response to the salt stress of both the wild-type and mutant genotypes, the activities of superoxide dismutase, catalase, and guaiacol peroxidase were unchanged, decreased, and increased, respectively. The JA-treated wild type plants showed the highest activity of all three enzymes as compared with the mutant plants. Salinity caused a decrease in chlorophyll content in the wild-type and jin 1 plants. Preliminary JA treatment of the Col-0 plants resulted in a normal content of photosynthetic pigments after the salt stress, while the positive JA effect was insignificant in the jin 1 mutants. It was concluded that the MYC2/JIN 1 protein is involved in the JA signal transduction and plant adaptation to salt stress.

Carnosine is an endogenous dipeptide with antiproliferative properties. Here we show that carnosine selectively inhibits proliferation of human glioblastoma cells (U-118-MG) compared to breast (MB231) and oral (Cal27 and FaDu) cancer cells. Carnosine-induced inhibition of U-118-MG proliferation is associated with a significant: decrease in cellular reactive oxygen species levels, increase in manganese superoxide dismutase (MnSOD) and increase in cyclin B1 expression resulting in G2-block. We conclude that the antiproliferative property of carnosine is due to its ability to enhance MnSOD and cyclin B1 expression. These results will be of significance to the potential application of carnosine in brain cancer therapy.

Evaluation of an antiviral effect of miRNA in the nanoparticles of a polycationic compound against mRNA of vp16 protein (UL48 gene) of herpes simplex virus type 2 (HSV-2) in vitro. MATERIALS AND METHODS. 50% aqueous solution of polyethyleneimine (BDH, Great Britain), chitosan, containing approximately 15% of N-acetylated glucosamine chains (Sonat, Russia), hydrazine-hydrate and other chemical reagents (Chimmed, Russia); Vero continuous cell line, MS HSV-2 virus were used. Vero cells were cultivated in DMEM medium supplemented by 10% fetal bovine serum at 37°C in the atmosphere of 5% CO2. Cell viability was evaluated by using Neutral Red vital stain and MTT-test. Primers and probes for RT-PCR were modeled in Vector NTI 8.0 computer program according to the mRNA sequences of the studied genes (the sequences were obtained from GenBank) and synthesized in Sintol (Russia). RT-PCR tests were set using a standard procedure. Synthesis of PEI-PG-chitosan was carried out by Krivtsov G.G. et al. (2010).

To study E-cadherin and β-catenin expression in colorectal cancer (CRC) liver metastases in order to assess the impact of different drug therapy regimens on the adhesive properties of tumor cells.

To evaluate the ameliorative effect of curcumin on dietary aflatoxin-induced changes in the expression of genes in Nile tilapia Oreochromis niloticus liver, the fish were fed with a diet contaminated by 200 ppb of atlatoxin B1 (AFB1) with and without curcumin (5 mg/kg diet) for 16 weeks in addition to a negative and positive controls fed with the basal diet and basal diet supplemented with curcumin, respectively. Further, two recovery groups with and without curcumin were tested after 2 more weeks. Relative mRNA expression of genes involved in antioxidant function (superoxide dismutase, SOD), biotransformation (cytochrome P4501A, CYPA) and immune response (interleukin-1β, IL-1β and transforming growth factor β, TGF-β) were assessed using RT-PCR. Also, fish weight gain and survival rate were determined. Results revealed that AFB1 significantly-reduced the survivability and weight gain, while curcumin inclusion improved them. Fish fed with AFB1-contaminated diet showed the up-regulation of CYP1A and down-regulation of SOD, IL-1β and TGF-β. This expression pattern was still evident in the recovery group without curcumin, but to a lesser extent. Supplementation of curcumin ameliorated the overall gene expression close to the control levels. It appears that curcumin exhibited protective effects on AFB1-induced liver toxicity in O. niloticus by moderating oxidative stress, toxin biotransformation, immune response, and hence growth performance.

We have investigated the role of apoptosis resistance gene bcl-2 in the activation of cellular senescence program induced by histone deacetylase inhibitor (HDACi) sodium butyrate (NaBut) in transformed rat fibroblasts. This study was conducted in a resistant to apoptosis induction cell line of rat embryo fibroblasts transfor- med by oncogenes E1A, cHa-ras and bcl-2 (ERasBcl). The parent cell line transformed with only EJA and cHa-ras (ERas) was used as a control. It has been found that NaBut reduces proliferation rate of ERasBcl cells significantly weaker than of ERas transformed cells, despite the fact that the G1 cell cycle arrest was observed in both cell lines. After NaBut treatment, hypertrophy of the apoptosis resistant transformants ERasBcl also was reduced compared to parent cell line ERas, due to less activation of mTORC1, which is known to control the synthesis of protein and ribosome biogenesis. The degree of mTORC1 activation was as.sessed by its target proteins phosphorylation: the ribosomal S6 protein and 4E-BP1--inhibitor of translation initiation factor eIF4E. Since cell senescence process may be associated with changes in autophagy regulation, we analyzed the dynamics of one of the main autophagosome formation markers--protein LC3. The accumulation of lipid-bound form LC3-II changes significantly in ERasBcl cells after NaBut treatment and has transient nature. The set of analyzed cellular senescence markers suggests that a high level of apoptosis resistance gene bcl-2 expression prevents the realization of tumor-suppressor senescence program induced by HDACi sodium butyrate treatment.

The expression of DDX5 protein (RNA-helicase p68) correlates with processes of proliferation and differentiation. However there is no direct evidence of involvement of the protein in these processes. In present work, we studied the influence of DDX5 protein inactivation by si-RNA on the proliferation of Jurkat cells and dynamic of DDX5 expression during differentiation of U-937 cells induced by phorbol 12-myristate-13-acetate (PMA). We showed that the content of DDX5 in Jurkat cells is less in phases G0/G1 as compare to phases G2/M. The treatment of cells with the antisense LV-shDDX5 was followed by the increase of G0/G1 cells. It was also shown that the increase of expression of the DDX5 protein occurred during the initial stages of differentiation, and the peak of expression was registered during the first 2-3 hours after the induction of the cells, later the DDX5 content decreases. The increase of the number of macrophage surface marker CR3 on the membrane of cells occurred only in 24 hours after induction of the cells by PMA. Thus, these data confirm that: (1) the DDX5 protein is essential for normal cell proliferation; (2) the transition from G1 to S/G2 phase is accompanied by an increase of DDX5 protein concentration in the cells; (3) the concentration of the DDX5 protein increases on early stages of U-937 cells differentiation and after decreases.

In order to evaluate the toxic effect of silver (AgNP) and titanium dioxide (TiO2) nanoparticles their influence on the expression of genes of biomarkers of inflammatory responses and apoptosis in human lymphocytes was studied. An increase in the IL-6, IL-8, TNF-α and p53 genes expression in the concentration range of silver and titanium dioxide nanoparticles of 10-40 μk g/ml was found. Increased expression of IL-6, IL-8, TNF-α and p53 genes under the nanoparticles action indicates the stimulation of the immune system and of apoptosis, respectively.

Hemolymph filtration in insects is performed by nephrocytes, additional cells of the circulatory system that are not connected to Malpighian vessels. Drosophila has two types of nephrocytes: the ventral ("garland"), which are situated around the connection site of the esophagus and proventriculus, and the pericardial, which are localized around the heart. In this study, we examined the role of the of insulin-like receptor (InR)gene in regulation of the function of ventral nephrocytes (VNC) in D. melanogaster females. Immunofluorescent analysis of female VNC with anti-InR antibodies revealed for the first time that the InR gene is expressed in VNC cells. To determine whether a change in the level of InR expression has an effect on VNC function in Drosophila females, we implemented an antisense suppressor of the InR gene, together with a driver that is expressed specifically in VNC. VNC function was evaluated by survival of the females exposed to toxic stress (treatment with AgNO3). This study has shown for the first time that suppression of InR expression in VNC leads to a rise in the survival of flies under conditions of toxic stress.

This paper studies the effect of plant peptides of thionine Ns-W2 extracted from seeds of fennel flower (Nigella sativa) and β-purothionine from wheat germs (Triticum kiharae), as well as a synthetic antimutagen (crown-compound), on the expression of several genes involved in the.control of cellular homeostasis, processes of carcinogenesis, and radiation response in human rhabdomyosarcoma cells (RD cells), T-lymphoblastoid cell line Jurkat, and blood cells. All of these agents acted as antimutagens-anticarcinogens, reducing the expression of genes involved in carcinogenesis (genes of families MMP, TIMP, and IAP and G-protein genes) in a tumor cell. A pronounced reduction in the mRNA level of these genes was caused by thionine Ns-W2, and the least effect was demonstrated by β-purothionine. Antimutagens had very little effect on the mRNA levels of the several studied genes in normal blood cells.

Comparison of action of chronic γ-irradiation at a dose of 22.6 cGy and the serpisten substance containing phitoecdysteroids at small doses of 5 and 50 mg/kg on biochemical indicators in erythrocytes and tissues of white not purebred mice is given. It is established that in both cases there is an increase of minor fractions of cardiolipin and phosphatidic acid, lysophosphatidilcholin and a share of phospholipids as part of common lipids. Course administration of serpisten to rats at the total doses of 12 and 30 mg/kg leads to an increase in tissues of thermal shock proteins of family 70 (Hsp70 and Hsc70). Similarity of action of ecdysteroid preparations and the influence of stress factors of physical nature of low intensity (gamma radiation at a small dose) have been detected in mice, which manifest themselves in some chain links of lipid peroxidation processes as well as an increase in biosynthesis of thermal shock proteins of family 70 (Hsp70 and Hsc70) in rats at administration of serpisten.