[The lysate and recombinant antigens in ELISA-test-systems for diagnostic of herpes simplex].

The lysate and recombinant antigens of various production included informula of ELISA-test-systems were analyzed. The ELISA-test-systems are used for detection of IgG to Herpes simplex virus type I and II. For testing the panel of serums PTH 201 (BBI Inc.) were used. The samples of this panel contain antibodies to Herpes simplex virus type I and II in mixed titers. The 69 serums of donors were used too (17 samples had IgG to Herpes simplex virus type I, 23 samples to Herpes simplex virus type II and 29 samples had no antibodies to Herpes simplex virus). The diagnostic capacity of mixture of recombinant antigens gG1 Herpes simplex virus type I and gG2 Herpes simplex virus type II (The research-and-production complex "DiaprofMed") was comparable with mixture of lysate antigen Herpes simplex virus type I and II (Membrane) EIE Antigen ("Virion Ltd."). In the test-systems for differentiation of IgG to Herpes simplex virus type I the recombinant antigen gG1 Herpes simplex virus type I proved to be comparable with commercial analogue Herpes simplex virus-1 gG1M ("Viral Therapeutics Inc."'). At the same time, capacity to detect IgG to Herpes simplex virus type II in recombinant protein gG2 Herpes simplex virus type II is significantly higher than in its analogue Herpes simplex virus-2 gG2c ("Viral Therapeutics Inc.").

[Simultaneous quantification of soluble fibrin and D-dimer in blood plasma for the assessment of the threat of thrombosis].

Soluble fibrin and D-dimer are the most specific markers of blood coagulation cascade and the threat of thrombosis. Two immunoassay test systems were designed using the fibrin-specific and D-dimer-specific monoclonal antibodies. The clinical trials of the test systems were carried out in Ukraine. The high informative value of soluble fibrin quantification as a prognostic indicator of the threat of thrombosis associated with hip replacement and endoprosthetics of the abdominal aorta was shown. Independent D-dimer quantification is less informative. Simultaneous quantification of soluble fibrin and D-dimer before operation and at different time intervals after it is required for the prediction of postoperative thrombotic complications and monitoring the efficiency of antithrombotic therapy. Only in this case it is possible to get information about the state of the balance between blood coagulation and fibrinolytic systems, and determine the degree of the threat of thrombosis.

[Production of purified human recombinant alpha2b-interferon].

Technology for producing biologically active recombinant alpha2b-interferon is based on creating a bacterial producer strain containing the cDNA of human interferon alpha. The authors have obtained two producers of recombinant alpha2b-interferon, the synthesis of the target protein in them occurs in the inclusion bodies. The schemes of isolation and purification of biologically active recombinant alpha2b-interferon have been developed. The drug purity was approximately 97-98%. Biological activity in the culture of sensory cells in the cytopathic test was 4.2*10(8)ME/mg.

[Obtaining of hairy-root, callus and suspension carrot culture (Daucus carota L.) able to accumulate human interferon alpha-2b].

Here we report the obtaining of suspension, callus and hairy root culture initiated from carrot plants of Nantskaya and Perfektzya variety with the highest level of recombinant human interferon alpha-2b accumulation exhibited the highest level of plant protein extract antiviral activity (up to 12.8 x 10(3) IU/mg TSP). The antiviral activity of callus extracts was significantly lower comparing to the activity of plant extracts from parent organisms. However, the antiviral activity level of suspension culture extracts (up to 4.42 x 10(3) IU/mg TSP) and Ri-root ones (up to 4.42 x 10(3) IU/mg TSP) appeared to be comparable to analogical data of antiviral activity of transgenic carrot leaf extracts, this way the described cultures could be possibly used for comparatively speedy obtaining of recombinant therapeutic protein for curing and preventing of virus diseases.

[Influence of physiological state of Escherichia coli cells on the expression of soluble protein--recombinant analog of glycoprotein G of Herpes simplex virus of type 2].

It is shown that the recombinant protein GST-HSV2gG, containing the immunodominant regions of glycoprotein G of HSV-2 is accumulated in the form of inclusion bodies or in soluble form in the Escherichia coli BL21 (DE3) cells. The ratio between protein fractions varied depending on the physiological state of cells before biosynthesis. The kinetic parameters of bacterial populations were determined by mathematical modeling of growth curves based on the Verhulst logistic function. It was established that the induction of biosynthesis in the growth acceleration phase (at OD600 = 0.3) with 0.1 mM IPTG gives the maximum yield of soluble protein (26.75 mg/l or 17.6 mg/g biomass). The target protein was purified using the immobilized metal ion affinity and affinity chromatography technologies. Antigenic activity of the soluble form of recombinant protein GST-HSV2gG, was significantly (three times) higher than that of the protein purified from inclusion bodies (p < 0.05) and was comparable with the activity of the commercial analog (p > 0.05), that allows using this product in the immunosorbent test kits for diagnosis of IgG to HSV-2.

[Nanocrystaline ceria based materials--perspectives for biomedical application].

Nanocrystalline ceria possesses a unique complex of physical and chemical properties making it highly bioactive material. In this review, modern data on the action of nanocrystalline ceria on cells, micro- and macroorganisms are analyzed. Special attention is paid to the analysis of the factors affecting protective properties of CeO2 with respect to the living systems.

[Antiviral effect of cerium dioxide nanoparticles stabilized by low-molecular polyacrylic acid].

Antiviral activity of nanosize cerium dioxide sols (CeO2) in animal cell culture has been studied. The inhibiting effect of the mentioned sols upon reproduction of vesicular stomatitis test-virus was demonstrated for the first time in case of preliminary 24-hour contact with cells lines L929 and EPT. The effectiveness of protective action depends on initial precursor and the way of obtained water nanosize nanosols.

[Thrombin inhibitors which display antibacterial activity].

The investigation of cytotoxicity and antibacterial activity of the novel thrombin inhibitors containing retro-D-sequences -D-Arg-D-Phe--modified by D-arginine amino group by the residues of lauric acid or chromone-contained substituent, in comparison with known cationic preservative Nalpha-lauroyl-L-arginine ethyl ester (LAE) have been carried out. It has been shown that compound Laur-D-Arg-D-Phe-OMe has a similar cytotoxicity with LAE, and Chrom-D-Arg-D-Phe-OMe has almost twice higher toxicity than it fatty moiety contained analogues. Antibacterial activity of Laur-D-Arg-D-Phe-OMe against Staphylococcus aureus and Bacillus subtilis is close in action to LAE. It is assumed that ability of thrombin inhibitors to suppress the growth of some microorganisms can be explained by their ability to suppress activity of trypsin-like serine proteinases, which participate in the infection process of Staphylococcus aureus and influence on Bacillus subtilis sporulation. These findings open new prospects for exploring efficient antimicrobial agents among synthetic low-molecular trypsin-like serine proteinase inhibitors.

[Gene-engineering technologies and stem cells].

In the review it has been presented the conceptional analysis of modern state of stem cells use problem (SC) in gene therapy ex vivo. Principles of vectorial systems construction have been expounded, designed for SC transfection. It has been displayed results of experimental investigations and clinical data on transplantation of genetically modifiable SC at different monogenous and multifactorial diseases. Special attention is focused on the problem of biosecurity ensuring.

[The role of Toll-like receptors in regulation of immune responses in norm and pathology].

We have analyzed the recent data about Toll-like receptors (TLRs) that play the most important role in host immune defense. TLRs involved in induction and modulation of innate and acquired immunity as the integrators of their reactions. Genetic disorders of TLRs or their signaling pathways components were noticed with different pathologies. TLRs are the ideal molecular target for the therapy of many diseases including inflammatory, autoimmune, allergic and tumor diseases.

[Interferon-stimulating activity of muramoyl dipeptide glycosides].

The uninvestigated interferon (IFN)-inducing capacity of glycoside derivatives of N-acetylmuramoyl-L-alanyl-D-isoglutamine (MDP) has been studied. Most MDP glycosides tested in vitro were more active than the reference preparation MDP. Under the in vivo conditions three studied preparations: MDP, a-heptyl-MDP and beta-(2-methyl-3-phenylchromonyl-7)-MDP stimulated production of considerable amount of circulating IFN that evidences for the promising character of their further investigation as preparations immunostimulators.

[Optimization of microbial synthesis of recombinant analogs of Treponema pallidum membrane proteins (Tpp17 and Tpp47) and estimation of their immunochemical properties].

Two strains which produce recombinant proteins--analogs of membrane immunodominant epitopes with molecular weights 17 and 47 kDa of Treponema pallidum were constructed on the basis of E. coli recipient BL21 (DE3)-Gold with phage T7RNA-polymerase expression system. The method of mathematical modeling of growth curve was used to determine kinetic parameters which characterized physiological state of the culture producer in the process of fermentation. Results of immunoenzymatic analysis of blood serum samples from patients with syphilis and healthy donors are presented.

[Mesenchymal progenitor cells. Biological characteristic and prospects for their use].

Imunophenotic characteristic of the mesenchymal stem and progenitor cells, their tissue origin and functional peculiarities as well as immunological aspects of its transplantation are presented in the article. Also the address migration and the site-specific differentiation of mesenchymal stem cells its age-related and pathological process induced changes are discussed. Prospects of clinical use of mesenchymal stem cells for the treatment of degenerative arthritis, spinal cord injuries, myocardial infarction and other disorders are shown.

[Functional activity of lymphoblastoid cells infected by human adenovirus type 2 and Epstein-Barr virus].

The paper deals with the influence of the adenovirus (Ad) and Epstein-Barr virus (EBV) on functional activity of lymphocytes, in particular, the production of alpha- and gamma-interferons, tumor necrosis factor (TNF) in conditions of mono- or double infection of B- and T-phenotype (CEM) lymphoblastoid cells. It is shown, that Ad, EBV or both viruses induce high enough levels of interferon on both lines of cells and in control epithelial cells. The lymphoblastoid cells infected by viruses deep ability to synthesize alpha- and gamma-interferons under the influence of the corresponding inducers (Newcastle disease virus and hemagglutinine). Nevertheless, the levels of their formation are not high. Rather high parameters of activity of the tumor necrosis factor (TNF) were revealed during a day in the initial B95-8 cells and superinfected Ad after the effect of LPS of E. coli. Their activity in CEM cells also did not depend on the infection type.

[Preparation of monoclonal antibodies to the Fc-fragment of human IgG and the use of their based immunoenzyme conjugates].

An original kit containing 17 clones of hybridomas, the producers of monoclonal antibodies (MCAb) against the Fc-fragments of human IgG, has been obtained. The possibility of using horseradish peroxidase conjugates of obtained MCAb as a part of ELISA test systems (Diaproph-Med, Ukraine) was comparatively studied. The application of peroxidase conjugates of two monoclonal antibodies (156C10 and 153H11) as part of five diagnostic kits was shown to provide a high specificity and a high sensitivity.

[Specific immunoglobulins as effective drugs for treatment of virus infections: present state and urgent tasks in Ukraine]. / Spetsificheskie immunoglobuliny kak éffektivnye preparaty dlia lecheniia virusnykh infekstii: sovremennoe sostoianie i nasushchnye zadachi dlia Ukrainy.

The thesis about importance of specific immunoglobulin preparations in treatment of infectious diseases of viral and microbial etiology, uncontrollable infections, caused by representatives of herpes viruses family in particular, is developed in the paper. The types of immunoglobulin preparations, their advantages, technology peculiarities has been considered; the experience of application, including that of the authors have been analyzed. The necessity and expedience of intensive development of this branch of immunobiotechnology in Ukraine is proved, especially in connection with a high level of infectious morbidity in this country, concrete recommendations are given.

[Effect of synthetic muramyl dipeptide derivatives on staphylococcal infection in mice]. / Vliianie sinteticheskikh proizvodnykh muramildipeptida na techenie stafilokokkovoi infektsii u myshei.

The work deals with the results of experimental evaluation of the influence of some new modified derivatives of muramyldipeptide (MDP) on the course of staphylococcal infection in mice. The preparations under study were found to produce rapid elimination of bacteria from kidneys and the increase of phagocytic activity of blood macrophages in animals. At the same time MDP and its derivatives stimulated natural killer cells whose activity was inhibited during infection. The dependence between the structure of these compounds and their protective action in staphylococcal infection, as well as the increase of the natural immunity characteristics of the body was followed.

[The antibacterial efficacy of preparations of interferon and its inducers]. / Antibakterial'naia éffektivnost' preparatov interferona i ego induktorov.

Experimental data have been presented which prove antibacterial efficiency of interferon preparations and inducers under infection processes evoked by Staphylococcus aureus, Salmonella typhimurium, Pseudomonas aeruginosa, Klebsiella pneumoniae. It is shown that the system of phagocytizing cells and natural cells-killers plays the basic protective role in the organism under the persistence of these microbes. The basic regularities of activating effect of interferon preparations and its inducers on the functional activity of the above mentioned cells have been found. The methods of treatment with interferon drugs based on experimental data have been developed for the first time. They were used to cure patients with pyo-septic diseases.