[In vitro study of antiviral activity of Myramistin against subtypes H3N2 and H5N1 of influenza virus].

In vitro antiviral effect of myramistin on influenza virus (MDCK cell culture) was studied. The drug showed significant dose-dependent antiviral activity against the virus. When used prophylactically (1 hour before exposure to the virus) in subtoxic doses, myramistin was effective in inhibiting replication of the influenza virus [strains A/Aichi/2/68 (H3N2) and A/Chicken/Suzdalka/Nov-11/2005 (H5N1)]. In urgent prophylaxis (1 hour after exposure to the virus) the protective effect was less pronounced, especially when the contamination dose was high. When the drug was added 12 hours after exposure to the virus, it had no protective effect on the MDSK cell monolayer. The prospects of the myramistin use as a prophylactic agent in grippe are discussed.

[Evaluation of the antimicrobial potential of immune sera].

The inhibiting activity of blood sera obtained from 20 volunteers immunized with the multi-component vaccine "PYOPOL", 25 nonimmune donors and 7 lots of human antistaphylococcal immunoglobulin with respect to Staphylococcus aureus, Escherichia coli, Proteus vulgaris and Pseudomonas aeruginosa has been studied in the liquid nutrient medium with the use an automated microbiological analyzer. The sera of donors immunized with the vaccine "PYOPOL" have been found to possess high antimicrobial activity, comparable with the definite concentration of the antiseptic agent "Myramistin". The proposed method may be recommended for the evaluation of the bacteriostatic activity of different immuno- biological preparations.

[In vitro study of antiviral activity of Myramistin against measles and mumps viruses].

The study was aimed at in vitro investigation of the Myramistin antiviral activity against the measles and mumps viruses in the Vero cell culture. The experiments with addition of myramistin simultaneously or at various periods after inoculation of the monolayer by the measles virus (Edmonson strain) or mumps virus (PetroNov/03 strain) revealed pronounced dose-dependent antiviral effect of the drug. It was shown that for prevention of replication of the measles and mumps viruses the optimal concentrations were 0.05 to 0.005%. The prospects of myramistin use as a prophylactic agent for infections caused by the measles and mumps viruses are discussed.

[Ways of overcoming drug resistance in patients with pulmonary tuberculosis]. / Puti preodoleniia lekarstvennoi rezistentnosti u bol'nykh tuberkulezom legkikh.

To elucidate the combined effect of surfactants and antituberculous drugs, the surfactant myrastimine was tested. Two series of sputum cultures with multidrug-resistant Mycobacterium tuberculosis (MBT) strains were made. The experimental results confirmed that myramistine noticeably enhanced the sensitivity of multidrug-resistant MBT strains to antituberculous agents. The use of endobronchial and intracavernous myramistine in combination with antituberculous agents to patients with destructive pulmonary tuberculosis caused by multidrug-resistant MBT strains reduced the latter's resistance to antibiotics, cured tubendobronchitis in most patients, created conditions for obliteration of the cavernous cavity. Positive changes are noted in the first 4 months after the initiation of therapy, which can prepare patients for surgical treatment more rapidly.

[Synthesis and biological activity of anomeric muramoyldipeptide butylglycosides]. / Sintez i biologicheskaia aktivnost anomernykh butilglikozidov muramoildipeptida.

The synthesis of anomeric butyl glycosides of muramyl dipeptide was reported. alpha-Butyl glycoside of N-acetyl-D-glucosamine was 4,6-O-benzylidenated and the benzylidene derivative was 3-O-alkylated by the Williamson reaction with sodium (S)-2-chloropropionate. The resulting protected alpha-butyl glycoside of muramic acid was then condensed with L-Ala-D-iGln-OBzl by the DCC-HOSu method. Mild acidic hydrolysis and subsequent catalytic hydrogenolysis of the resulting glycopeptide yielded the target alpha-butyl glycoside of N-acetyl-L-alanyl-D-isoglutamine. In the synthesis of beta-butyl glycoside of N-acetylmuramyl-L-alanyl-D-isoglutamine, 2-acetamido- 4,6-di-O-acetyl-2-deoxy-3-O-[(R)-1-(methoxycarbonyl)ethyl]-alpha- D-glucopyranose, a 1-OH derivative of muramic acid, was the key compound. Its interaction with the excess thionyl chloride resulted in the corresponding glycosyl halide, which was condensed with n-butanol according to Helferich. O-Deacetylation, 4,6-isopropylidenation, and subsequent alkaline hydrolysis of the resulting compound gave the protected beta-butyl glycoside of muramic acid. Its activation and condensation with L-Ala-D-iGln-OBzl and the subsequent removal of protective groups were performed in the same manner as the reactions in the synthesis of alpha-butyl glycoside of N-acetyl-L-alanyl-D-isoglutamine. The adjuvant activity of the butyl glycosides to HIV proteins rgp160 and rgp120 and their ability to affect in vitro HIV replication and the proliferation of mouse spleen T-cells were examined. The biological activity of anomeric muramyl dipeptides was shown to depend essentially on the configuration of their anomeric center.

[A modification to the chloroform-methanol method of isolating the matrix protein of the influenza virus]. / Modifikatsiia khloroform-metanol'nogo metoda vydeleniia matriksnogo belka virusa grippa.

A modification of chloroform-methanol method of isolation of M1 protein (M1P) of human influenza A virus allowed increasing 1.6 times the yield of the target product and decreasing 6.4 times the amount of organic solvents. It has been shown that M1 protein isolated by the modified method did not differ (as to its physicochemical characteristics and antigenic activity) from M1 protein isolated by means of the initial method.

[Surface-active substances in the arsenal of agents against AIDS]. / Poverkhnostno-aktivnye veshchestva v arsenale sredstv bor'by so SPIDom.

Only nucleoside analogues such as zidovudine have been widely used to improve the clinical state of patients with AIDS till now. Their use has a number of eliminations. Nevertheless, the anti-HIV properties of many different substances have been shown. Some of them contain hydrophobic and polar parts of molecules and can be considered as surfactants. Detergents (DT) (among them Ukrainian preparation Myramistin), modified phospholipids, polyene antibiotics (such as Nystatin A), lipophilic muramyl peptides (LMP), saponins (S) and some other surfactants have been described as inhibitors of HIV replication. Moreover, LMP and S are the potent adjuvants and are probable candidates for vaccines against HIV. Adjuvant activities of some DT have been shown recently as well. We hope that elaboration of new adjuvants with strong anti-HIV properties is an attractive and manageable aim.

[Effect of saponins from Hedera taurica Carr. and modified muramylpeptides on replication of human immunodeficiency virus in vitro]. / Vliianie saponinov iz Hedera taurica Carr. i modifitsirovannykh muramilpeptidov na reproduktsiiu virusa immunodefitsita cheloveka in vitro.

The effects of muramyldipeptide (MDP), several new MDP derivatives, and saponins derived from Hedera taurica Carr, on the in vitro replication of HIV-1 were studied. The coefficient of alteration of the rate of HIV replication was used to compare these reagents' effects on virus replication in Jurkat-tat cells. This coefficient was calculated as the ratio of concentrations of HIV p24 in supernatants to the amount of viable cells. Muramylpeptides boosted HIV replication. Only one modified muramylpeptide alpha-butyl-MDP and tauroside H2 were not capable of boosting HIV-1 antigen production.

[The use of monoclonal antibodies and a biotin-streptavidin system of amplification for the quantitative determination of human IgE]. / Ispol'zovanie monoklonal'nykh antitel i biotin-streptavidinovoi sistemy usileniia dlia kolichestvennogo opredeleniia IgE cheloveka.

A "sandwich"-variant of the solid phase immunoenzyme method of quantitative determination of human IgE in biological liquids has been developed on the basis of two monoclonal antibodies to various epitopes of Fc-fragment of E-chain of human IgE and biotin-streptavidin system of the signal amplification. The total analysis time is 3 h: the range of determined concentrations of IgE is 1-200 kE/l; sensitivity-1 kE/l; the type of calibrating curve is linear. To make an analysis twice 10 microliters of the tested sample are required. Reproducibility of the suggested method is not below 5%.

[Optimization of a method of production of allergen extracts from ambrosia pollen]. / Optimizatsiia metoda polucheniia allergennykh ékstraktov iz pyl'tsy ambrozii.

Peculiarities of allergens extraction from the intact and destructed pollen grains of Ambrosia artemisiifolia L. have been studied, and the effect of the content of extracting buffer on stability of the obtained allergen extracts has been investigated. It is established that mechanical destruction of pollen during extraction permits decreasing protein output, increasing specific activity of the extracts and accelerating the procedure of their production. Introduction of some protective additions to the composition of the extracting buffer permits essentially increasing stability of the extracts under long-term storage at 4 degrees C.

[The use of the immunoperoxidase test with monoclonal antibodies for detecting influenza A viruses]. / Ispol'zovanie immunoperoksidaznogo testa s monoklonal'nymi antitelami dlia vyiavleniia virusov grippa A.

The immunoperoxidase test was used to detect influenza virus in cells of a chorionallanthois shell of infected chicken embryos. Application of monoclonal antibodies D8 and A11 in the analysis has permitted detecting reproduction of type A (subtypes H1N1, H2N2, H3N2) viruses, the PGA titre of the respective allantois liquids being not lower than 1:16. The matrix protein and hemagglutinin, detection of which underlies this analysis, were found on the cell membrane, in the perinucleus region and as cytoplasmic inclusions.

[The effect of fetal calf serum on the anti-HIV properties of miramistin]. / Vliianie émbrional'noi syvorotki korov na anti-VICh-svoistva miramistina.

The anti-HIV activity of an antiseptic miramistin in the presence of fetal calf serum (FCS) was studied. It has been recently shown that cation detergent miramistin at a concentration of 100 micrograms/ml prevented HIV-1 replication in MT-4 cells when these cells were cocultivated with MT-4 cells previously infected and treated with the detergent in a protein-free medium. Under such conditions 50 micrograms of miramistin per ml delayed HIV propagation in MT-4 cells by 14 days. Detergent-dependent arrest of HIV development was abolished by addition of FCS at a final concentration of 50%. Miramistin in a dose of 100 micrograms/ml prevents HIV-1 replication in Jurkat-tat cells when they are cocultivated with the cells of the same line previously infected and treated by the detergent in the presence of 5% FCS. FCS in concentrations from 12.5 to 75% prevents the anti-HIV activity of miramistin in a dose of 100 micrograms/ml.

[The use of monoclonal antibodies for detecting the influenza virus]. / Ispol'zovanie monoklonal'nykh antitel dlia vyiavleniia virusa grippa.

The possibilities of using influenza A (Leningrad) 385/80 (H3N2) virus matrix protein-specific FITC-labeled D8 monoclonal antibodies in immunofluorescence assays were investigated. The virus antigen accumulation was detected in chorioallantoic cells of chick embryos. Exhibiting the type-specific properties, the fluorescent antibodies stain the perinuclear space, cytoplasmic membrane, and granular structures in the cytoplasm of infected cells. The haemagglutination test tires in the corresponding specimens were at least 1:16.

[The induction of alloantigen-specific cytotoxic T-lymphocytes by the intravenous immunization of mice]. / Induktsiia alloantigen-spetsificheskikh tsitotoksicheskikh T-limfotsitov pri vnutrivennoi immunizatsii myshei.

It is shown possible to induce specific cytotoxic T-lymphocytes (CTL) in the course of intravenous (i.v.) immunization of Balb/c mice by 2000-rad-irradiated allogeneic C57Bl/6 splenocytes in a dose of 9 x 10(7). The induced CTL express Thy1.2+L3T4-Ly2+ cell surface markers. No correlation was observed between the level of cytotoxic activity and the ability to inhibit proliferation in the population of lymphocytes primed by i.v. immunization.

[The stimulation of the proliferative reaction of immune splenocytes by monoclonal antibodies to the beta-chain of the LFA-1 molecule can lead to an increase in their cytotoxicity]. / Stimuliatsiia proliferativnoi reaktsii immunnykh splenotsitov monoklonal'nymi antitelami k beta-tsepi molekuly LFA-1 mozhet privodit' k usileniiu ikh tsitotoksichnosti.

A system for investigation of relationship between proliferation and cytotoxicity of immune splenocytes fractions stimulated by monoclonal antibodies (mAb) to beta-chain LFA-1 molecule was elaborated. To obtain different proportional contents of effector cells in populations, the immune splenocytes were eluted from donor, third-party, and recipient macrophage monolayers. Cytotoxic indices of effector splenocyte fractions were compared before and after their proliferation in mixed lymphocyte culture. The results do not exclude the role of proliferation in increasing the effector cells proportions in populations stimulated by mAb to beta-chain LFA-1 molecule.

[Monoclonal antibodies to the beta-chain of the LFA-1 molecule promote an increase in the cytotoxic index of effector lymphocytes and stimulate their proliferation]. / Monoklonal'nye antitela k beta-tsepi molekuly LFA-1 sposobstvuiut povysheniiu tsitotoksicheskogo indeksa éffektornykh limfotsitov i stimuliruiut ikh proliferatsiiu.

The influence of monoclonal antibodies (mAb) to LFA-1 molecule beta-chain on the functional activities of immune splenocyte fractions which were eluted from donor, third party and recipient macrophage monolayers was studied. The splenocyte fractions were treated with the mAb and tested for cytotoxicity and proliferation. MAb to LFA-1 molecule beta-chain (without co-stimulation by other antibodies) were found to stimulate the immune response of the splenocyte fractions in both functional tests. The observed effect might be non-specific. The relationship between the stimulation of proliferative and cytotoxic responses of the immune splenocyte fractions and the treatment with mAb of LFA-I molecule beta-chain remains obscure.

[Effect of the Ly2 molecule on the function of alloantigen-specific effector cytotoxic T-lymphocytes as a function of the variability of their receptors' affinity]. / Vliianie molekuly Ly2 na funktsiiu alloantigenspetsifichnykh éffektornykh tsitotoksicheskikh T-limfotsitov v zavisimosti ot variabel'nosti srodstva ikh retseptorov.

To study T cell receptors' affinity alloantigen-specific anti-K cytotoxic T-lymphocytes (CTL) were divided on fractions by elution from donor K and third-party macrophage monolayers. The functional activity of CTL was suppressed by anti--Ly2 monoclonal antibodies (mAb), 51-Cr-labelled transformed fibroblasts (L-cells) transferred with the H-2K gene were used as targets. The results demonstrate that primary CTL enriched on third-party macrophage monolayers are the most sensitive to anti-Ly2 mAb. T-cells exhausted on third-party monolayer and then enriched on donor monolayer were resistant to treatment with the mAb. Secondary CTL enriched on donor monolayer were resistant to treatment with anti-Ly2 mAb even should they were not exhausted on third-party monolayer. These results show that Ly2 (CD8) molecule plays an essential role in the interaction of CTL with MHC class I molecule only if T-cell receptor has low affinity.

[Biosensors in biotechnology and medicine]. / Biosensory v biotekhnologii i meditsine.

The work is a review of the state-of-art of research in the field of biological sensors. The enzymatic and immune biosensors are described. Problems concerning application of microorganisms as biosensors have been reported in detail. The use of luminescent bacteria and luciferase sensors is under discussion. The principles of operation of fibrous-optic and semiconducting biosensors are stated.

[Monoclonal antibodies to the influenza A virus matrix protein that react with viral strains of different subtypes]. / Monoklonal'nye antitela k matriksnomu belku virusa grippa A, reagiruiushchie s virusnymi shtammami razlichnykh podtipov.

To obtain hybridomas secreting monoclonal antibodies (MCA) to matrix (M) protein of influenza virus, mice were immunized with a modified antigen which consisted of subvirus units after electrophoretic removal of surface glycoproteins from virions using Desintegnon-O detergent. Six stable hybrid cultures producing MCA to M-protein were derived. The properties of MCA to the antigen determinant common for a group of influenza A virus strains of different subtypes allow them to be used in the development of a test-system for diagnosis of influenza A type.