The Inhibitory Activity of Lactobacillus plantarum Supernatant against Enterobacteria, Campylobacter, and Tumor Cells.

Cell-free supernatant of Lactobacillus plantarum exhibit a strong antimicrobial effect against a number of pathogenic enterobacteria (E. coli, Shigella flexneri, Salmonella typhimurium, Proteus mirabilis, and Campylobacter jejuni). The degree of growth inhibition in broth culture reached a high level for all tested bacteria. The highest rates were noted for P. mirabilis (by 13 times) and the lowest for S. flexneri (by 5 times) and C. jejuni (by 4.5 times). Significant antiproliferative effect of the supernatant on cells of tumor-derived epithelial cell lines was shown. The highest degree of inhibition (by 22 times) was observed for HT-29 cells (colon carcinoma). Thus, inclusion of probiotics in traditional treatment schemes can increase the effectiveness of antibacterial and antitumor drug therapy.

[Interferon-regulating activity of the CelAgrip drug and its influence on the formation of reactive oxygen species and expression of innate immunity genes in Burkitt's lymphome cells cultures.]

INTRODUCTION: Interferons (IFN) and IFN inducers are effective in suppressing viral reproduction and correcting of the innate immunity mechanisms. The aim of the study was to test the hypothesis of the possible involvement of the IFN inducer CelAgrip (CA) as an activator or suppressor of antiviral effects in Burkitt's lymphoma (LB) cell cultures with different ability to produce Epstein-Barr virus antigens (EBV). MATERIAL AND METHODS: The kinetic analysis of the dynamics of reactive oxygen species (ROS) production and determination of gene group expression by real-time PCR in response to CA treatment were done in human cell lines LB P3HR-1 and Namalva, spontaneously producing and not producing EBV antigens. RESULTS AND DISCUSSION: When treating CA in Namalva cells, a decrease in the ROS activation index was found; in P3HR-1 cells, an increase was observed. After treatment with CA, there was no reliable activation of the IFN-α, IFN-ß and IFN-λ genes in Namalva cells, but the expression of the ISG15 and P53(TP53) genes was increased more than 1200 times and 4.5 times, respectively. When processing the CA of P3HR-1 cells, the expression of IFN-α genes increased by more than 200 times, IFN-λ - 100 times, and the ISG15 gene - 2.2 times. The relationship between IFN-inducing action of CA and the activity of ISG15 and ROS in LB cell cultures producing and not producing EBV antigens is supposed. CONCLUSION: In Namalva cells that do not produce EBV antigens the treatment of CA results in suppression of ROS generation and activation of the expression of genes ISG15 and P53 (TP53); in P3HR-1 cells producing EBV antigens, the opposite picture is observed - the formation of ROS and the expression of the IFN-α and IFN-λ genes are activated and the activity of the ISG15 and P53 (TP53) genes is suppressed.

[Interferon-regulating activity of the celagrip antiviral drug and its influence on formation of reactive oxygen species and expression of innate immunity genes in the follicular lymphoma patients].

INTRODUCTION: Medicines from the group of interferon inducers (IFNs) "swith on" the synthesis of type 1 interferons (IFN-I) and induce the expression of IFN-stimulated genes (ISGs) that regulate innate immunity reactions and protect the host from infectious agents and the tumour pathology.The purpose of the study was to determine the role of the drug celagrip (CA) in the activation of innate immunity genes and the effect on the production of reactive oxygen species (ROS) in patients with follicular lymphoma (FL). OBJECTIVES:  to study the intensity of ROS production and the level of expression of the IFN-α2, IFN-λ1, ISG15, BCL2, P53(TP53) and USP18 genes in response to the treatment of blood cells of patients with FL with the preparation of CA. MATERIAL AND METHODS: The study involved primary cancer patients diagnosed with follicular lymphoma (FL) and healthy volunteers. A kinetic analysis of the dynamics of production of reactive oxygen species (ROS) was performed in whose blood cells, and the expression of the group of genes was determined by real-time PCR in response to CA processing. RESULTS AND DISCUSSION: ROS production by blood cells of patients with FL and volunteers in the presence of CA significantly decreased (P < 0.05). The level of gene expression of ISG15, P53(TR53) and USP 18 in the group of patients with FL was significantly higher than that in the group of volunteers. When treating blood cells with CA, it becomes possible to divide patients with FL into groups with a positive and negative response in accordance with the level of expression of the USP18 gene. We divided FL patients into groups with a positive and negative response in accordance with the level of USP18 gene expression after treatment of blood cells with CA. CONCLUSIONS: The CA drug reduces the production of ROS and simultaneously stimulates the activity of the innate immunity genes ISG15, P53(TP53) and USP18 in the blood cells of patients with FL.

[Cytokine-regulating activity of anti-virus preparation CelAgripus in Burkitt's lymphoma stable B-cell lines.]

INTRODUCTION: Cytokines activated in response to immunosuppressive viral infections can directly or indirectly affect the neoplastic transformation of B cells. In this study, we studied a new substance designed to produce the antiviral drug CelAgrip (CA, CelAgripus), which exhibits interferon (IFN) and cytokine-inducing activity and, apparently, can be used as an activator of antiviral immunity. Purpose - is to evaluate the cytokine-regulating effect of CA in Burkitt's lymphoma (LB) cell lines latently infected with the Epstein-Barr virus (EBV). OBJECTIVES: to study the CA-induced expression of the cytokine genes IL-1ß, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-17, IL-18, IFN-α, IFN -γ, IFN-ß, IFN-λ1, IFN-λ2, IFN-λ3, TNF-α in normal and EBV transformed LB cells. MATERIAL AND METHODS: Cell line: the human embryo fibroblasts (HEF), Namalva, Daudi, Raji, P3HR-1. Preparations: CA, gossypol-acetic acid (GAA), sodium carboxymethyl cellulose (Na-CMC). METHODS: RT-PCR and methods for assessing cytotoxicity (MTT and Scepter 2.0 Merck cell counter). RESULTS: The effect of the CA preparation on the expression of IFN-λ, IL-1ß, IL-6, IL-8 and IL-10 genes was revealed. DISCUSSION: We observed the activation of gene expression of IFN-λ, IL-1ß, IL-6, IL-8 and suppression of IL-10 gene activity when treatment CA of LB cells. CONCLUSION: The substance CA has new effects on the activation of the expression of a number of key cytokine genes in stable Burkitt lymphoma cell lines.

Genotoxicity of single-walled carbon nanotubes: in vitro study on human embryonic fibroblast cells.

The effects of single-walled carbon nanotubes on the levels of DNA aberrations, chromosome and genome disorders were studied on human embryonic fibroblasts, their karyotype was analyzed by the spectral karyotyping method. The level of DNA aberrations increased after 3-h exposure to the nanotubes. No appreciable increase in the incidence of aberrant metaphases, micronuclei, and chromosome 1, 6, 8, 11, X, and Y aneuploidy after 24- and 48-h incubation with the nanotubes were detected.

[The interaction between nerve cells and carbon nanotube networks made by CVD process investigation].

In this research we investigate neuroblastoma cells cultivated on single-walled carbon nanotubes networks made by CVD method on silicon substrates. The complex analysis of grown cells made by atomic force, electron microscopy and Raman spectroscopy was carried out and the effect of nanotube growth process on proliferation factor was investigated. It is shown that despite of a weak decrease in proliferation, cell morphology remains unchanged and no physical or chemical interaction between carbon nanotubes and cells is observed. The results of the research can be used to investigate the interaction between conductive nanomaterials and cells for the development of neural replacement implants. Also they can be useful in bio-electronic interface investigation of signal propagation in neurons.

[Cultivation of continuous cell lines on the substrate of carbon nanotubes and the effect of electric stimulation on cell proliferation].

It was demonstrated that the three studied samples of carbon nanotubes of domestic production fixed on the substrate surface did not have toxic effect and could be used for cell cultivation. A biocompatible conductive coating based on carbon nanotubes and bovine serum albumin was developed. The efficacy of the coating for growing in vitro cell cultures was tested. A device was developed for electric stimulation of the cells. Local electric potential was applied to the cells using nanoscale electrodes. The results of human embryonic fibroblast cultivation in a pulsed electric field on conductive nanocomposite substrates were presented. An 26% increase in the proliferative activity of cells was observed at potentials up to 100 mV.

[Regularities in the spread of hepatitis C virus and its genotypes in Russian and countries within the former USSR]. / Zakonomernosti rasprostraneniia virusa gepatita C i ego genotipov v Rossii i stranakh SNG.

The incidence of markers of hepatitis C virus (HCV in the blood of 4216 normal subjects living in the European Russia (Northern, North-Western, Central, Central Chernozem, Volga-Vyatka, Volga, and North Caucasian regions), in the Urals, in Siberia (Eastern Siberian region), in the Far East, and in Monogolia is assessed. The incidence of antibodies to HCV varied from 0.7% in the Central region to 3.8% in the Central Chernozem and 10.7% in Mongolia. HCV genotyping (identification of 1a, 1b, 2a, 2b, and 3a genotypes) was performed using 469 RNA of HCV-positive sera of donors and patients collected in Russia, Moldova, Turkmenistan, and Mongolia. The 1b genotype predominated everywhere (68.9%), its incidence being the highest in Moldova (96%). Unclassifiable variants of HCV were found in 28 (6%) of sera. The regularities of HCV genotypes circulation in the European Russia were the same as in other European countries, whereas their prevalence in Eastern Russia was rather like that in China or Japan. The prevalence of genotypes did not depend on the clinical manifestations of diseases caused by HCV.

Prevalence of hepatitis C virus and distribution of its genotypes in Northern Eurasia.

We tested hepatitis C virus (HCV) antibody in 4216 sera collected from healthy people living in European part of Russia (including Northern, North-Western, Central, Central-Blacksoil, Volga-Vyatka, Volga, and North-Caucasian regions), non-European part of Russia (the Urals, East-Siberia, and the Far-East regions) and Mongolia. Prevalence of HCV antibody varied significantly by regions, ranging from 0.7% in Central region of European part of Russia to 10.7% in Mongolia. Genotyping of HCV (into 1a, 1b, 2a, 2b, and 3a) was performed on 469 sera from blood donors and patients (in Russia, Moldova, Turkmenistan, and Mongolia) who were positive for both HCV antibody and RNA. Genotype 1b was the most dominant genotype irrespective of regions (68.9%), with the highest rate in Moldova (96%). HCV unclassifiable into genotypes 1a-to-3a was found in 28 (6.0%) samples: particularly 4 of 10 samples from Lipetzk were untypable. Overall, HCV genotypes in European part of Russia were more similar to those in European countries, while those in Eastern part of Russia more similar to China or Japan. Genotype distribution was not associated with the clinical expression of HCV disease: acute hepatitis, chronic hepatitis or liver cirrhosis.

[The experimental efficacy of vaccination with a recombinant yeast vaccine against hepatitis B in combination with immunomodulators]. / Effektivnost' v éksperimente vaktsinatsii rekombinantnoi drozhzhevoi vaktsinoi protiv gepatita B v sochetanii s immunomoduliatorami.

Experiments in BALB/c mice were carried out to study a number of immunomodulators in combination with vaccination using a recombinant yeast hepatitis B vaccine. The use of poludan, ridostine, amixine, larifan, myekonide significantly enhanced specific antibody production and at least doubled cell-mediated immune response by 14 and 28 days postvaccination. The above-mentioned immunomodulators are planned to be used in vaccination with the recombinant yeast vaccine against hepatitis B.

[Proteins induced by various types of double-stranded RNA and interferon type I in human fibroblasts. Correlation with antiviral activity of the substances]. / Belki, indutsirovannye raznymi vidami dvispiral'nykh RNK i interferonami tipa I v fibroblastakh cheloveka. Korreliatsiia s antivirusnoi aktivnost'iu preparatov.

Protein induction by new antiviral preparations of dsRNAs (larifan, ridostin, rifastin and poly(A).poly(U)) and recombinant beta-interferon in human fibroblasts (M19) was studied. The common gene products: 88, 80, 68, 58, 56, 52, 50 and 26 kD were detected in the spectra of the induced cytoplasmic polypeptides. At the same time the sets of the induced proteins had individual distinctions in various preparations. Induction of the 56-kD protein was more essential in the action of dsRNAs than that of interferon. The antiviral activity of dsRNAs and interferon preparations correlated with a relative increase in the synthesis of proteins with molecular weights of 88, 80 and 58 kD. The study results are in agreement with the fact that the dsRNAs have interferon-independent pathways of antiviral action with participation of 56- and 58-kD protein genes.

[Enzyme activity of interferon system in virus diseases]. / Aktivnost' fermentov sistemy interferona pri virusnykh zabolevaniiakh.

The enzymes activities of interferon system at viral infections of different etiology/influenza, parainfluenza, hepatitis B with delta infection and urticaria chronica with respiratory and herpes infection/has been studied and evaluation of the noted changes in the enzymes activity is discussed. It is shown that the same pathological changes in enzymes activities of interferon system were observed at different viral infections.

[Levels of interferon-dependent enzymes in urticaria patients, connected with frequent viral infections]. / Urovni interferonzavisimykh fermentov u bol'nykh krapivnitsei, sviazannoi s chastymi virusnymi infektsiiami.

27 patients with relapsing and giant urticaria accompanied by multiple general respiratory viral infection and herpetism were studied during acute and remittent steps as compared with 13 healthy volunteers. 10 patients were treated with reaferon. Elevated rates of circulating interferon and interferon-dependent enzymes as well as a decreased ability of leukocytes and lymphocytes to produce alpha- and gamma-interferons were detected in these patients. Anomalous reactions of the interferon system were found in the patients with urticaria and general viral respiratory infection simultaneously with impairments of the immunity response: discorrelation between rates of interferon-dependent enzymes and circulating interferon. The course of interferon therapy within 5 days using recombinant alpha 2-interferon did not obviate these deteriorations, although activity of 2-5A synthetase tended to decrease down to the level observed in healthy persons.

[Activation of the interferon system enzymes by recombinant alpha2-interferon in patients with chronic hepatitis B]. / Aktivatsiia rekombinantnym alpha2-interferonom fermentov sistemy interferona u bol'nykh khronicheskim gepatitom B.

Treatment of patients suffering from chronic hepatitis B with recombinant leukocytic interferon (reaferon) increased the levels of circulating interferon and activated interferon-dependent enzymes such as 2-5A-synthetase and histone kinase. Activation of the enzymes was observed for 1 to 2 weeks. After that period it was maintained at the required levels with intramuscular administration of 1-3 million units of reaferon 2 or 3 times a week. In parallel with increasing of the levels of the interferon system enzymes there was observed a decrease in the level of aminotransferase. The reaction of the viral antigens to the treatment with reaferon was not the same: HBe antigen and antibodies to HBe antigen disappeared, the content of HBs antigen and antibodies to delta-interferon did not change.

[Interferon status and the activity of dsRNA-dependent enzymes in volunteers and influenza patients receiving human recombinant alpha 2-interferon (reaferon)]. / Interferonovyi status i aktivnost' dsRNK-zavisimykh fermentov u dobrovol'tsev i bol'nykh grippom liudei, poluchavshikh chelovecheskii rekombinantnyi al'fa 2-interferon (reaferon).

The national recombinant preparation of alpha 2-interferon (reaferon) after intramuscular inoculation in a daily dose of 3 X 10(6) units/ml to human volunteers was found in the blood serum within 1-2 hours. In 85% of the subjects given the preparation, a 2-8-fold rise in interferon titers in the blood serum and in 64% an increase in interferon leukocyte reaction (ILR) by the 5th day after the beginning of the treatment were observed. In the examined influenza patients given the preparation, interferon circulation in the blood serum, increased ILR and 2-5A synthetase enzyme levels were demonstrated. Tests for interferon of the blood sera of patients given reaferon by inhalation revealed no significant rise in its level, in contrast to cases with reaferon intramuscular injections. It is assumed that the immunomodulating effect of the preparation given by inhalation is mediated through the hypothalamus. Thus, parameters of the interferon system may be corrected by administration of reaferon.

[Morphogenetic characteristics of the mammary glands of autoimmune F1 mice (NZB x NZW)]. / Osobennosti morfogeneza molochnykh zhelez autoimmunnykh myshei F1 (NZB x NZW).

The morphogenesis of mammary glands was studied in the normal and autoimmune F1(NZW X NZB) mice. In the lactation cycle of the autoimmune mice the normal course of structural-functional rearrangements of parenchyma and stroma in the developing and involuting mammary glands was disturbed. A conclusion has been reached that the modification of stromal elements, first of all involved in the autoimmune disease, is the leading link in the abnormal development of mammary glands.

[Formation of transfecting DNA in acute cellular infection with the tick-borne encephalitis virus mediated by an oncogenic virus]. / Obazovanie transfitsiruiushchei DNK pri ostroi infektsii kletok virusom kleshchevogo éntsefalita, oposredovannoe onkogennym virusom.

To prove the association of the transfecting activity of nucleic acid from cells co-infected with TBE and SV40 viruses with DNA, experiments were carried out using fractionation of nucleic acid preparations in cesium sulphate gradient an on a column with HAP. The experiments led to a conclusion that the infectivity of the preparations used was associated with double-stranded DNA. Investigation of nucleic acid infectivity form cytochalasine-enucleated cells indicated that the function of the nucleus was necessary for formation of transfecting DNA in mixed TBE and SV40 infection of cell. No formation of transfecting DNA was observed in experiments of blocking replication and transcription of cellular genome. The retention of the transfecting activity of DNA during 6 passages of cells at approximately the same level indicated the lack of selective preferences or inhibition of cells containing transfecting DNA.

[Morphophysiological characteristics of the development and involution of mammary gland structures in murine radiation chimeras]. / Morfofiziologicheskie osobennosti razvitiia i involiutsii struktur molochnykh zhelez myshinykh radiatsionnykh khimer.

A study of development and involution of the mammary glands was carried out in syngeneic and xenogeneic mouse radiation chaimeras at different times after transplantation of the bone marrow cells, before and after restoration of the lymphoid tissue. The data obtained suggest the important role of the lymphoid (immune) component in the processes of development and involution of the mammary gland structures, as well the necessity of genetic homology of the hemopoietic elements and the mammary gland cells.