[Development and properties of neutralizing monoclonal antibodies for fusion protein of respiratory syncytial virus.]

INTRODUCTION: Respiratory syncytial virus (RSV) is the most common cause of lower respiratory tract infections in infants and the elderly. The absence of a wide range of therapeutic drugs and vaccines indicates to the high relevance of the development of new effective drugs for the prevention and treatment of RSV infections. PURPOSE: to obtain highly active and specific monoclonal antibodies (MAbs) capable of detecting RSV in infected cells and neutralizing the infectious activity of the virus in vitro. MATERIAL AND METHODS: RSV reference strains of group A 2 subgroups (A2 and Long) were propagated in HEp-2 and MA-104 cell lines, respectively. Mice were immunized with purified RSV A2 virus. MAbs were obtained using hybridoma technology. RESULTS: A panel of 6 MAbs reacting with RSV strains А2 and Long has been obtained. Four MAbs were IgG (IgG2a or IgG2b subtype), two MAbs were IgM. All MAbs reacted with RSV F-protein in immunochemical tests. The MAbs actively reacted with RSV in ELISA, in immufluorescence and peroxidase staining of infected cells, and in immunodot test. Five out of 6 MAbs neutralized of RSV in cell culture. Different properties of MAbs suggest that they target different antigenic sites of F-protein. DISCUSSION: Comparative analysis suggests that the obtained MAbs can be used for the development of diagnostic preparations, for RSV detection in clinical materials and confirmation of infection etiology by rapid culture method. CONCLUSION: High activity and specificity of MAbs indicate that they can serve as a basis for development vaccines and preventive medicines.

Resistance of THP-1 Leukemia Cells Infected with Cytomegalovirus to Anti-tumor Antibiotic Doxorubicin and Restoration of the Sensitivity by Inhibitors of the PI3K/AKT/mTOR Molecular Pathway.

Results obtained showed that infection with HCMV prevented the death of THP-1 cells treated with DOX in both active and latent forms of infection. In the presence of mTOR inhibitors (rapamycin and Torin2), the sensitivity of the infected cells to DOX was restored. Rapamycin inhibited the expression of the HCMV protein IE1-p72 and increased sensitivity to DOX. Molecular targets for the creation of new drugs for the treatment of leukemia in patients infected with HCMV were determined.

Insecticides of neonicotinoides class: determining exposure via workers' urine.

For biomonitoring of exposure in workers with insectocaricides, the authors created a method of multi-component assessment oflowlevels of neonicotinomides in workers' urine, based on last generation tandem liquid mass-spectrometry (triple quadrupole) with ionization source - electrostatic dispersion (positive ionization) in dynamic multi-reaction monitoring with two transitions of parent ions (for quantitative assessment and ionic ratio confirmation). After the work, the operators gave urine samples (about 100 ml in average) that were frozen and kept under -20°C before analysis. Samples were defrozen before analysis, and each urine portion of 5 ml was diluted by equal volume of 0,1% formic acid. To extract substances out of the samples, solid-phase extraction (cartridges based on octadecylsilane) was applied, elution was performed with 1 ml of methanol. Lower limit of the substances detection in urine - 0,02-0,05 ng/ml, lower limit of the quantitative assessment - 0,1-0,2 ng/ml. The method was tested on monitoring of the workers' exposure to preparations based on imidaclopride and clotianidine in natural conditions of pesticides use in agriculture with various processing technologies. midacloprid was identified in urine of 3 professional operators after wheat and oat seeds treatment and after subsequent seeding at lower limit of detection (0,02 ng/ml), lower limit of quantitative assessment (0,1 ng/ ml) and 0,34 ng/ml.

[Biomonitoring in workers exposed to pesticides: development and application of method detecting imidacloprid in urine].

Imidacloprid is a relatively new insecticide in neonicotinoids chemical class with neuroactivity in insects, being one of the most widely used insecticides in the world. For biomonitoring in workers exposed to pesticides, the authors designed a method detecting low levels of Imidacloprid in urine of operators, based on tandem liquid mass-spectrometry with ionization source--electrostatic dispersion (positive ionization) in multi-reaction monitoring regime with subsidiary ion (mass/charge) 209 for quantitative assessment and ion 175.1 for confirmation onion ratio. The study incorporated diurnal urine, about 100 ml of average sample was frozen and kept at temperature -20C for analysis. Before extraction, the sample was unfrozen, an aliquot of 5 ml was selected, diluted with 5 ml of 0.1% formic acid. The substance was concentrated out of the urine samples via solid-phase extraction with application of cartridges based on octadecylsilane, eluition--1 ml of methanol. Lower limit of Imidacloprid detection in urine is 0.02 ng/ml, of the quantitative assessment--0.1 ng/ ml, linear range of concentrations measured 0.1-10 ng/ml. The method was tested for monitoring in workers exposed to Imidacloprid preparations in natural conditions of pesticides application in agriculture, with various processing technologies. Imidacloprid was identified in urine of two professional operators after work in seed treatment and the subsequent seeding at lower limit of detection (0.02 ng/ml) and 0.34 ng/ml.

[The method of multicomponent determination of herbicides of various chemical classes in water].

In the work there are considered results of the development of the multicomponent method of measurement of concentration of herbicides of various chemical nature under their joint presence in the water. There was justified the optimality of application of HPLC-DAD (the working wavelength of 240 nm) for the determination of levels of 10 active ingredients of herbicides of class of sulfonylurea (metsulfuron-methyl, nikosulfuron, sulfometuron-methyl, thifensulfuron-methyl, triflusulfuron-methyl), imidazolinone (imazapyr, imazethapyr), 2,6-Bis[(4,6-dimethoxy-2- pyrimidinyl)oxy]benzoic acid (bispyribac acid), triazol-pyrimidines (Penoxsulam), a benzoylpyrazole compound (Topramenzone). For the concentrating and cleaning of samples of water there were used cartridges for solid-phase extraction of Oasis HLB - the macro porous copolymer made on the basis of the balanced ratio of 2 monomers - lipophilic divinylbenzene and hydrophilic N-vinylpirrolidone. The range of the detected concentrations in water was volatile between 0.0005 and 0.005 mg/L, values of standard deviation vary in the range of 1.8-3.9%. Chlorine-containing acidic herbicides were analyzed by the method of GC-ECD and GC-MS (IE) after preliminary converting of compounds into flying derivatives with the use of diazomethane. Satisfactory extraction of substances from a water sample may be achieved by classical extraction in the system "liquid-liquid" with the application of Methyl tert-butyl ester. For cleaning of the derivatized sample there were used cartridges for solid-phase extraction on the basis of silica gel. The range of the determination of 9 active ingredients referring to classes of phenoxy-acetic acid (2,4- D, MCPA), pyridinecarboxylic (aminopyiralid, picloram, clopyralid), benzoic acids (dicamba), benzothiadiazinone (bentazone), biphenyl ester (acifluorfen) and a chloroacetamide (acetochlor) - 0.0001-0.001 mg/L, SD values vary in the range of 1.8-33%.

[Determination of residual amounts of chlorothalonil in peaches: problems of gas chromatographic identification with the use of electron capture detector].

In the work there are presented results of studies on the validation of the gas chromatographic (GC) methodfor the determination of chlorothalonil residue amounts in peaches with the use of electron capture detector (ECD). For the analytical control there was selected such stone fruit crop as the peach, referring to the crops, the most contaminated with residue amounts ofpesticides. There was justified the necessity of the inclusion in the procedure of the detection of the method of confirmation, based on mass spectrometry detection (MSD) (the type of ionization - electron impact). The significant source of the obtaining of incorrect data in the identification with the use of ECD of ions are shown to be phthalates, visualized in the chromatogram as intense and/or broad peaks. Mass spectra of compounds of the class ofphthalates are characterized by the dominant peak of the ion with the value of (mass/ charge) 149, just on this peak the detection of low molecular weight phthalates occurs in various matrices, on the spectrum there are also recorded typical ions corresponding to fragments of radical residues. The combination of the use of various types of the detection allows to prove that the revealed response (detector signal) is caused just by the analyte, but not the impurities, and optionally to optimize chromatographic conditions towards to the obtaining reliable results. The lower limit of the quantitation of chlorothalonil in peach fruits accounts for 0.01 mg/kg, determined with a signal/noise ratio of 10. The range of measured concentrations is volatile between 0.01-0.1 mg/kg, recovery rate of chlorothalonil from samples of peaches, established according to results of the analysis of model samples with the introduction of the substance in four points along the detection range, was 84-102%, the SD value of the repeatability varies in the range of 2.0-5.8%.

[EFFECT OF ANTI-CANCER DRUG DOXORUBICINE ON CYTOMEGALOVIRUS INFECTED HUMAN FIBROBLASTS].

The anticancer antibiotic doxorubicine (DOX) is highly toxic and induces functional complications in vital organs. The effect of DOX on normal cells has not been examined in sufficient detail, and the search for compounds reducing DOX toxicity did not lead to success so far. It has been suggested that DOX induces death of cancer cells via p53-dependent apoptosis, however, the information regarding the role of p73 protein, a member of p53 tumor suppressor family, is scanty. Cytomegalovirus (CMV) induces an antiapoptosis program that allows its replication until death of the target cell. Our objectives were to examine the effect of DOX on normal cells (human fibroblasts), analyze the ability of CMV-induced antiapoptosis program to reduce DOX toxicity, and to evaluate the involvement of p73 protein and its isoforms in the regulation of death of CMV-infected and DOX-treated cells. Within a 24-h time period DOX caused death of about 70% human embryonic lung fibroblasts (HELF) in cell culture, this parameter decreased significantly in CMV-infected DOX-treated HELF cells. TUNEL has shown that the number of cells with DNA fragmentation decreases from 5.2% under the effect of DOX to 3.2% (P < 0.05) after combined CMV-DOX treatment. Analysis of mitotic figures revealed that DOX causes accumulation of mitotic cells, which was not observed in CMV-infected DOX-treated cells. PCR analysis of mRNA of two p73 protein isoforms (TAp73 and dNp73) has shown that in uninfected cells the expression of TAp73 isoform was low, while in CMV-infected cells level of TAp73 was significant and expression of dNp73 was demonstrated for the first time. Expression of TAp73 associated with lack of mitosis block. The activation of caspases 8, 9 and 3 in CMV-infected cells was registered but cell death was not, however, as massive as that caused by DOX. From these findings it can be concluded that CMV attenuates DOX-related damage to normal cells. It can be suggested that induction of TAp73 and dNp73 isoforms provides conditions for reduction of DOX effect which leads to DNA damage and death of normal cells.

[The development of diagnostics of herpes viral infections].

The representatives of herpesviruses family (Herpesviridae--virus of herpes simplex and human cytomegalovirus) are largely widespread in human population. These herpesviruses bring on severe disorders of embryonic development up to fetal death, newborn diseases, neurologic disturbances, deafness, and blindness and in transplantation patients the severe internal organs affections and transplanted organs rejection. Both herpesviruses are able to affect the central nervous system and result in encephalitis with lethal outcome. The particular attention deserve the asymptomatic forms in case of which virus is excretee and can be transmitted both by horizontal line (sexual way included) and by vertical line (in the process of itrauterine development offtetus). The lacking of clear djfefrentiating clinical symtoms frequentyv observed under manifestations of herpesviruses infections brings another danger The comparison is made of fouur methods of detection of herpes simplex in urethra scrapes: the immunocytochemical method of detection of herpes simplex antigen in smears: the rapid cultural method; the immune-enzyme method of detection of herpes simplex antigen; the polymerase chain reaction. The results demonstrated that the rapid cultural method detected herpes simplex in infected samples on all stages of disease and in even more quantity at the stage of exacerbtion as compared with the polymerase chain reaction. The presented data testify the actuality of development of of laboratory diagnostics of herpesviruses injection to relevant diagnosis, determination of form and stage of disease, timely initiation of treatment and monitoring of therapy.

[Methodical aspects of hygienic safety assessment of polymeric materials in contact with foodstuffs].

It was purposed new technique by capillary gas chromatography (GC) for the low level determination of monomer hexamethylenediamine (HMDA) in food simulants water from polymeric materials in contact with foodstuffs. Hexamethylenediamine, HN2-(CH2)6-NH2, is a monomer used in the manufacture of certain of polyamide plastic materials and articles intended to come into contact with foodstuffs. Compound exhibits all the chemical properties of aliphatic amines, is an irritant, causing dermatitis, can accumulate in the body, the degree of human exposure to HMDA assigned to the 2nd class of hazard - the substance is highly dangerous. There was studied two methodological approaches pre-derivatization of compound for GC determination. The first approach involves conversion of the free diamine using ethyl chloroformate as derivatizing agent followed by analysis of the resulting diurethan by gas chromatography using a flame ionization and mass selective detection (HMDA was quantitated by selective ion monitoring at m/z 102, the lower detection limit of 1 ng). According to second methodological approach the water samples were mixed with sodium chloride and extracted with toluene, then derivatized with trifluoroacetic anhydride (60 min, 55 degrees C) to diamide, 1 M potassium phosphate buffer (pH 7,0) was add to remove excess derivatizing agent, followed by analysis of resulting diamide by gas chromatography with electron capture detection (lower limit value 0,01 ng). Conformation of HMDA levels is carried out by combined gas chromatography/mass spectrometry (HMDA was quantitated by selective ion monitoring at m/z 126, the lower limit value of 0,1 ng). The optimal pre-derivatization of the second approach for the determination of low levels of HMDA in the water extracts. The range of measured concentrations of 0,005-0,5 mg/dm3, recovery 88-101%, the total error of measurement is 16%, the relative standard deviation is 1,85%. The method was tested in the study of aqueous extracts of the 10 random samples intended for food purchased in the consumer market. Shows the corresponding output level hexamethylenediamine requirements for products of this type.

[Different regulation of mitochondrial apoptosis and Bcl-2 gene expression in quescent and proliferative human fibroblasts infected with cytomegalovirus].

The aim of the study was to compare the dynamics ofmitochondrial apoptosis (MA) in cells at different stages of proliferation and with different susceptibility to cytomegalovirus (CMV). It has been found that in quiescent human fibroblasts (HF) CMV regulates MA at the level of bcl-2 gene transcription, exerting both pro- and anti-apoptotic effects. Suppression of bcl-2 transcription is greater in HF-977 line, which is highly susceptible to CMV in comparison with HF-1068 line. The effect of proliferative activity on MA was studied using CMV-infected HF-110044 line at the G0- or S-phase. A direct correlation was established between accumulation of cytochrome c and caspase 3 (MA markers) and production of IE72, pp65 and gB (CMV proteins). In G0-fibrob-lasts, viral replication was highly productive and bcl-2 expression was 10-fold as high as in S-phase cells, in which viral protein production and cell death were much lower. The increased gene transcription and accumulation of Bcl-2 protein enhanced cell viability and provided synthesis of viral proteins. Impaired structure of actin microfilaments, a caspase 3 target, coincided with pronounced suppression of gamma-actin gene in S-phase HF-110044. Our findings provide an insight into CMV-induced mechanisms of MA which lead to rapid death of infected quiescent fibroblasts and to slow death of cells infected at the stage of DNA synthesis.

[Localization of human cytomegalovirus immediate-early protein IE-1 pp72 in the juxtanuclear inclusion at the late stage of infection].

Immediate-early protein IE-1 pp72 is one of the most abundant proteins at the early stage of human cytomegalovirus infection and has a number of intranuclear activities. This paper gives immunocytochemical and ultrastructural data on IE-1 pp72 accumulation in the juxtanuclear inclusion at the late stage of low-multiplicity infection. Detection of a new localization site infers that this protein may participate in the final steps of virus morphogenesis and play a functional role in the pathogenesis of cytomegalovirus infection.

[Effect of Panavir on Herpes simplex virus types 1 and 2 proteins synthesis in cell culture].

The effect of Panavir on the synthesis of ultraearly (alpha), early (beta) and late (gamma) proteins of Herpes simplex virus types 1 and 2 in the culture of Vero cells was studied. It was shown that the level of the proteins suppression depended on the infection multiplicity and the time of the Panavir addition to the culture.

[Monoclonal antibodies to highly pathogenic avian influenza A(H5N1) strain isolated in the Russian Federation: development and properties].

Highly pathogenic avian influenza (HPAI) virus subtype H5N1 has recently caused extensive epizootics in different regions of the world and presents a serious threat to man. Since 2005, HPAI virus subtype H5N1 strains have been circulating in Russia, which differ from the earlier isolated Southern Asia strains. A panel from 15 monoclonal antibodies (Mabs) to HPAI virus A/duck/Novosibirsk/56/05 (H5N1) was developed. Eleven Mabs interacted with the hemagglutinin molecule (HA), 4 with influenza A virus nucleoprotein (NP) in the Western blot assay. The bulk of the obtained Mabs interacted with homologous virus in ELISA and showed an antigen in the infected cells in the indirect immunofluorescence assay. Nine Mabs were active in the hemagglutination inhibition (HI) assay and 8 of them were capable to neutralize viral activity. The comparative analysis of the properties of Mabs in the HI assay using various influenza A strains showed that Mabs 2C6, 6F3, 4G10, 3G9, and 7B3 inhibited hemagglutination of study avian influenza viruses subtype H5, Mab 6F3 being most active. Mab 3B5 reacted only with the viruses isolated in the Russian Federation in 2005-2007 and failed to interact with the other study influenza A viruses subtype H5. The obtained panel of Mabs can be used to study the fine antigenic structure of hemagglutinin and to make a differential diagnosis of avian influenza viruses subtype A/H5N1. The high neutralizing activity of Mabs creates a prospect for preparing humanized antibodies for specific prevention and treatment.

[CMV-induced cell death and fas gene expression in resting and proliferating human fibroblasts].

Cytomegalovirus (CMV) infection development and mRNA fas transcription levels (CD95) in resting (GO) and proliferating (S-phase) human lung embryo fibroblasts (HLEF-110044 line) were studied. In GO cells accumulation of infectious CMV was high and cell death was very quick, and fas gene expression was inhibited in early period of infection. In cells infected during S-phase CMV synthesis was lower and total cell death was detected only after 5 days; fas gene activity remained on high levels and increased during 6-48 hours. Death of CMV-infected fibroblasts occurred through apoptosis with cytopathic effect and detachment of cells in early stage, but without changes of cell membrane permeability and internucleosome fragmentation of DNA during later stages. In another HLEF-977 line CMV-induced apoptosis correlated with increased levels of fas gene transcription in resting cells. Positive association of activation Fas-receptor pathway and cell proliferation as well as different effect of CMV on activity of fas gene in 2 HLEF lines are discussed.

[Influence of membrane-active polyanions on various stages of the human cytomegalovirus life cycle in vitro].

Human cytomegalovirus (CMV), an agent of infection (CMVI), lethally dangerous for immune deficient neonates and adults was investigated in vitro as a target for a therapeutic effect of new membrane-active polyanionic compounds (MPC). Previous studies on the alicycle- and sulfate-modified carboxy-MPCs revealed a well-defined tendency of the anti-CMV activity amplification in parallel with increasing of the content of sulfate groups, enhancing the negative charge of the macromolecule. The dominating role of the electrostatic factor was confirmed by the highest activity of AS-688, compound with maximum sulfation among the tested MPCs. Its selectivity index (SI) of the CMVI inhibition in human diploid fibroblast cells reached 5450, 7500, 250 and 4286 in the microbicidal, viricidal, prophylactic and therapeutic schemes of the experiment respectively. The antiviral activity at the first, second and third schemes was explained by the polyanion-typical potential of electrostatic neutralization of the countercharged virions and prevention of the virus adsorption on the cell membranes (in competition with heparin sulfate, a cellular receptor of CMV), whereas the therapeutic effect required the ability of MPC to influence the intracellular stages of the CMV life cycle. The PCR and immunochemical assays revealed an inhibitory action of AS-688 on replication of the viral DNA and the following synthesis of the late viral protein gB with efficiency similar to that of gancyclovir (GCV). However, in contrast to GCV, acting as inhibitor of enzyme (viral RNA-polymerase) factor of the biosynthesis, the therapeutic activity of MPC could be interpreted by competition with viral RNA/DNA due to the specific character of the MPC molecular basis, initially constructed on the principle of nucleic acids backbone and charge adjustable imitation. This mechanism assuming reduction of the cytotoxicity risks, explained the experimentally observed fact of low cytotoxicity of MPCs and possible achievement of high SI. The MPC ability to penetrate into the cells without disruption of cellular membrane permeability was confirmed in experiments with the fluorescent-labeled derivate AS-679, structurally and functionally related to AS-688. In the light of the previously described HIV inhibiting properties of AS-688, AS-679 and MPC analogous, the results could be considered prospective in development of new highly effective agents for combined antiviral protection.

[Antiviral activity of polycarboxylic substances modified by cage hydrocarbon and sulfoacid pharmacophors against cytomegalovirus infection in vitro].

Human cytomegalovirus (CMV) infection (CMVI) results in lethal risks at the immunodeficiency status, including the HIV co-infection. Carboxy-mimickers of the polymeric backbone of nucleic acids, potential agonists and antagonists of the virus genome were developed as promising candidates for the antiviral protective agents. In parallel with stimulation of antiviral immunity the mimickers derived membrane potent compounds (MPC), were shown to be able to prevent directly and efficiently the cell infection by various strains of the human immunodeficiency virus (HIV) [Antibiotics and Chemother 2003; 48: 2:29-41; 5:7-15]. The paper presents new data and discussion of the results on investigation of the MPC, modified by the previously designed adamantane or norbornene and by the recently applied sulfoacidic pharmacophores in the experimental model of CMVI in vitro (human diploid fibroblast cells). Eight substances with various ratios of theabove mentioned cage-hydrocarbon and/or anion pharmacophores in the macromolecule were tested and active MPC modifications were detected which efficiently inhibited the CMVI with high indexes of selectivity up to 250, 4286 and 7500 in prophylactic, therapeutic and viricidal experimental schemes respectively. Modulating influence of the lipotropic (cage-hydrocarbon) pharmacophores on the anti-CMV activity was observed only in the viricidal and prophylactic experimental schemes, in which the lipid membranes of the cells and/or virus envelopes were involved. Still, the dominant role in the antiviral activity of MPC in all the experimental schemes was played by the sulfoacid-anionic chemical structure modulation. By increasing the density of the negative charge of the macromolecules to the levels comparable with the charge of the genome molecules, theanionic modification evidently amplified the potential of the antagonistic competition of the synthetic MPC with the virus genome, thus impairing the virus-specific interactions. The most promising compounds AS-688 and AS-678/-679 were selected for further investigation of the mechanisms of the anti-CMV and anti-HIV activity.

[Regulation of human fibrolastic 2',5'-oligoadenylate synthetase gene activity in cytomegalovirus infected fibroblast cells].

Great differences were found in the level of 2',5'-oligoadenylate synthetase 40-46 kDa (OAS1) mRNA in relation to the proliferative state of human diploid fibroblasts at the moment of cytomegalovirus (CMV) (the strain AD169) infection. In the phase of synthesis of cellular cycle DNA (S), CMV induced OAS1 mRNA transcription by 10-100 times stronger than then in phase Go infection. The level of viral induction OAS1 mRNA peaked by hour 12 postinfection. The high gene activity correlated with suppressed DNA synthesis, a slowing-down mitotic cycle, markedly inhibited CMV replication, and delayed cell death. When the cells were infected in phase Go, the stimulation of OAS1 gene activity was less and it was attended by intensive viral replication and rapid cell death. There was a direct relationship between the resistance of cells and the constitutive level of OAS1 gene expression: in the low CMV-sensitive cells, the activity of OAS1 gene was more than 10 times greater than that in the highly sensitive cells. The inhibitors of the enzymatic OAS activity induced by IFN and dsRNA were found in the cytoplasm of the CMV-infected cells.

[Carboxy mimetic derivatives of nucleic acid polymeric backbones inhibiting human cytomegalovirus. 1. In vitro microbicidal effect].

The artificial polycarboxyacidic compounds (PC), imitating the principle of furan-derived and negatively charged structures alternating in the polymeric backbone of nucleic acids, previously explored as interferon inductors and stimulators of antiviral immunity in vivo, were modified by the side groups to amplify the direct antiviral potency in vitro and investigated in the cell culture model of human diploid fibroblasts infected with human cytomegalovirus (CMV) in a microbicidal scheme. Reconstruction from the PC to membrane potent compounds (MPC) was carried out by covalent modification with lipotropic pharmacophores (of cage-hydrocarbon structures similar to rimantadine or camphor-like terpenoids), as well as by conversion of the carboxy groups to sulfate-anionic derivates, related to the CMV sensitive heparansulfate receptor (HSR) of the cells. Both the factors of the MPC structure-functional modulation (lipotropic and anionic) were found to be effective tools for amplification of the microbicidal activity. The maximum inhibitory effect against CMV and minimum cytotoxicity (with the best selectivity, the chemotherapeutic index of > or = 3000-5000) were achieved mainly through increasing the anionic groups content, elevating the MPC negative charge to the level comparable with one of the like charged viral genome and HSR. In relation with the previously found anti-HIV efficacy of the same MPCs in analogous experimental models and in view of the fact that CMV is one of the most dangerous opportunistic co-factors of HIV/AIDS pathogenesis, the obtained data can be used as a basis for further development of new generation microbicides, promising for combined prevention of sexually transmitted infections.

[Detection of the markers of herpes simplex virus and cytomegalovirus in newborns and infants].

A total of 111 children suspected for herpesvirus infection were examined. In blood and urine samples the infectious activity of herpes simplex virus (HSV) and cytomegalovirus (CMV) was detected by the rapid culture method (RCM) and the presence of virus DNA--by the polymerase chain reaction (PCR). HSV and/or CMV were detected by two laboratory methods in 57 examined children (51%). Of these, in 18 children (16.2%) both HSV and CMV were detected. The coincidence of the results of the detection of HSV and CMV by these two methods was observed in 72.4% and 75.2% of cases respectively. The comparative analysis of the detection of anti-CMV IgG and IgM was made with the use of commercial test systems produced bythe following manufacturers: "Vector-Best" and "Bioservice" (Russia), "HUMAN" and "Boehringer" (Germany). The effective detection of both anti-CMV (IgG and IgM) was ensured by the test systems "Boehringer". The test system "Vector-Best" for anti-CMV IgG proved to be not inferior as regards sensitivity and specificity. The German test systems demonstrated the highest specificity in the detection of low-avid antibodies. The data obtained in this study indicate that the detection rate of HSV and CMV markers in newborns and infants suspected for herpesvirus infection was, on the average, 20 - 40%. Reliable diagnostics in newborns and infants is possible only in the presence of the combination of at least 2 serological tests (the determination of antivirus IgM and IgG avidity) and 2 methods for the detection of direct herpesvirus markers (PCR and RCM).

Antiviral activity of liposomal preparations of antibiotic geliomycin.

A liposomal preparation with maximally possible content of incorporated geliomycin was obtained. Its cytotoxicity was studied in a culture of human embryonal diploid fibroblasts. Antiviral activity was studied on a model of cytomegaloviral infection in vitro by the capacity to plaque formation. Liposomal geliomycin was 10-fold less toxic for human cells than the solution of the antibiotic in DMSO and exhibited antiviral activity towards cytomegaloviral infection at a concentration of 0.042 microg/ml.