[Treatment of surgical stress response by neuroleptics, leu-enkephalin, glycyl-proline dipeptide, and electroanesthesia in experimental animals].

We have studied the effects of chlorpromazine, acepromazine, droperidol, and transcranial electroanesthesia upon evacuation function of the stomach in piglets and the effects of leu-enkephalin and glycyl-proline upon secretory activity of the stomach in dogs and rats during surgical stress to optimize anesthetic dosage. All pharmaceutical and nonpharmaceutical methods of anesthesia used in the experiments were implemented by actiovating stress-limiting systems in post-operational period. Leu-enkephalin, apart from stimulating the stress-limiting factors of neurohumoral systems of the organism, shifted the vector ratio of aggressive and protective factors of gastric mucosa toward strengthening of the latter. The opposite trends of changes in the secretory activity of the stomach of rats in response to surgical stress can be leveled using glycyl-proline as a component of anesthesia.

Optimization of parameters of physiological systems in animals under conditions of experimental stress exposure.

Physiological and biomedical experiments were performed. We estimated the parameters of tree main levels of physiological systems as a response of an organism to stress. Mechanisms of physiological defense of an organism against stress should be considered systemically. The response of physiological systems of animal organism was modeled by the activation of latent functional reserves by various exposures to stress (abdominal surgery, cranial electrotherapy stimulation, cranial electrotherapy stimulation with simultaneous administration of droperidol, aminazine, vetranquil, and carbacholine).

[Quantitative determination of the infective activity of the human immunodeficiency virus by plaque formation using monolayer cultures]. / Kolichestvennoe opredelenie infektsionnoi aktivnosti virusa immunodefitsita cheloveka metodom bliashkoobrazovaniia s ispol'zovaniem monosloinykh kul'tur.

A method for assessment of the infective activity of HIV using plaque formation method with human diploid cells (strains L-65, L-68, L-72) has been developed. This method is not inferior to plaque formation with poly-L-lysine in sensitivity, but superior to it in reproducibility of results and standardization of the test conditions. The method is fit for titration of HIV and virus-neutralizing antibodies, as well as for assessment of the activity of anti-HIV preparations.

[Study of the anti-HIV activity of miramistin]. / Izuchenie anti-VICh-aktivnosti miramistina.

The anti-HIV activity of a cationic detergent myramistin and nonionic detergent dezintegron-0 (d-0) was studied using HIV-1 strains III B/H9 and BRU in lymphoblastoid cells MT-4 and Jurkat-tat. Myramistin in a concentration of 0.075 mg/ml was shown to prevent HIV-1 replication in MT-4 when these cells were cocultivated with the cells preinfected and treated with the detergent. Myramistin in concentrations from 0.030 to 0.050 mg/ml delayed the accumulation of virus antigens in the cells by 4 and 14 days, respectively, without preventing the infection of intact cells. Nonionic d-0 prevented HIV-1 infection of intact Jurkat-tat cells at a concentration of 12.5 mg/ml.

[The isolation of HIV from children with the initial signs of an HIV infection and from symptom-free carriers]. / Vydelenie VICh ot detei s nachal'nymi priznakami VICh-infektsii i bessimptomnykh nositelei.

A high rate of HIV carrier state was observed in seropositive children with early symptoms of HIV infection. The virus was also isolated from 2 seropositive adults (mothers) showing no clinical manifestations. The intervals of virus manifestation in culture varied from 6 to 30 days with maximal frequency of detection in the 2nd week. Different modifications of the procedure for HIV isolation were assayed, and it was shown that the efficacy of isolation (shortening of the period of virus detectability and increase in the number of the antigen-containing cells) could be improved by the addition to the culture of the Jurkat-tat III line expressing the product of the tat gene important for virus reproduction.

[The strain features of the HIV-1 circulating in the USSR based on data from a study of its properties in cell cultures]. / O shtammovykh osobennostiakh tsirkuliruiushchego v SSSR VICh-1 po dannym izucheniia ego svoistv v kletochnykh kul'turakh.

Both variants of HIV-1 reported in the literature: slow/low and rapid/high types, were detected among the strains isolated from the subjects examined in 4 foci of HIV-1 infection in the south of the RSFSR and Byelorussia. All the 17 strains isolated in the southern RSFSR foci belonged to the slow/low type and had a low and unstable replication potential in donor peripheral blood mononuclear cells and in MT-4 cell line. All of them were isolated from subjects with asymptomatic infection and from children with initial clinical manifestations of the disease. Only one strain isolated in Byelorussia belonged to the rapid/high type. Its replicative activity was very similar to that of the classical HIV-1--HTLV-IIIB strain. Long-term (up to 7 months) propagation of slow/low strains did not result in any increase of their replicative activity. The capacity to form syncytia was found not only in the rapid/high type strains but also in the majority of slow/low strains under study.

[Dependence of alkaline phosphatase activity in human cultured fibroblasts on the proliferative status of the culture]. / Zavisimost' aktivnosti' shchelochnoi fosfatazy kul'tiviruemykh chelovecheskikh fibroblastov ot proliferativnogo statusa kul'tury.

The alkaline phosphatase activity was studied in embryonic lung human fibroblasts grown in vitro. The total enzyme activity per flask was constant during the log-phase of culture growth to increase significantly after confluence. The increase in the activity did not depend on the rate of culture growth, and was seen to start after the mean density equal to (37.5 +/- 5.9) X 10(3) cells/cm2 flask surface. The stimulation of cell growth may presumably slow down the synthesis of alkaline phosphatase, which resumes after the contact inhibition of DNA replication.

[Phase-specific protein synthesis in human fibroblasts after synchronization by a double thymidine block]. / Fazospetsificheskii sintez belkov v fibroblastakh cheloveka posle sinkhronizatsii dvoinym timidinovym blokom.

Kinetics of acid-insoluble non-histone protein synthesis during S and G2 cell cycle phases of diploid human fibroblasts and heteroploid transformed cells was investigated. Two distinct groups of protein with different kinetic pattern depending on the cell culture type were revealed. Export of one group of protein and turnover of the other group of acid-insoluble non-histone protein is arrested in heteroploid transformed cells.

[Isolation and characteristics of a new strain of diploid cells from human embryonic lung tissue]. / Poluchenie i kharakteristika novogo shtamma diploidnykh kletok iz émbrional'noi tkani legkogo cheloveka.

A new strain of the embryonic human fibroblasts L-68 was obtained and thoroughly characterized. It completely met all the requirements of the International Committee on the Cells Cultures. This strain can be recommended as a substrate for production of viral vaccines, diagnostic preparations and for research purposes.

[Glucose metabolism and template synthesis in the mitotic cycle of human diploid fibroblasts]. / Metabolizm gliukozy i matrichnye sintezy v mitoticheskom tsikle diploidnykh fibroblastov cheloveka.

The correlation between the rates of protein and nucleic acid synthesis and the activity of the key enzymes of glycolysis (hexokinase, phosphofructokinase) and pentose phosphate cycle (glucose-6-phosphate dehydrogenase) in the mitotic cycle of human diploid fibroblasts synchronized by double thymidine block was studied. It was found that the removal of the thymidine block is followed by short-term (presumably, non-specific) simultaneous stimulation of matrix syntheses, as well as by glycolytic and pentose phosphate cycle enzyme syntheses. By the beginning of the S-phase, all the processes appear to be inhibited, followed by gradual activation of glycolysis and pentose phosphate cycle reactions. The implementation of the cell cycle is concomitant with stepwise transitions of protein and hexokinase synthesis rates and ATP content to one of the following levels--basal, intermediate or maximal. Changes in the activity of glucose-6-phosphate dehydrogenase in the course of the cell cycle appear as oscillations, those in phosphofructokinase as alternative states. At stage M, the oscillatory processes are temporarily quenched, whereas the ATP content occupies an intermediate level. In contrast with diploid fibroblasts, in transformed T9 cells the enzyme activity is much higher, and the fluctuations in activity throughout the cell cycle are less noticeable. Presumably, in transformed cells the enzyme activity is at the maximum level and is not prone to effector regulation.

[Sequence of activation of template biosynthesis in normal and transformed human cells after synchronization by a double thymidine block]. / Posledovatel'nost' aktivatsii matrichnykh biosintezov v normal'nykh i transformirovannykh kletkakh cheloveka posle sinkhronizatsii dvoinym timidinovym blokom.

The sequence of matrix biosyntheses of DNA, RNA and various proteins in normal and transformed human fibroblasts in the first mitotic cycle after synchronization of cells by double thymidine block was studied. Two important regularities of synthesis of acid-soluble histone-like and acid-insoluble proteins in normal and transformed cells were established. In normal fibroblasts, the synthesis of both acid-soluble and acid-insoluble proteins is minimal before DNA replication and maximal in the G2-phase; that in transformed cells is maximal after removal of the thymidine block and decreased in the G2-phase. In normal fibroblasts, the synthesis of acid-insoluble proteins is maximal before, while that of acid-soluble ones--after the maximum of DNA synthesis. In transformed cells the situation is opposite. RNA synthesis in normal and transformed cells is stimulated at the end of the G2-phase. In normal cells, protein synthesis is coupled with the activation of RNA synthesis, whereas in transformed fibroblasts protein synthesis occurs, in all probability, in the next mitotic cycle. These differences are especially well-pronounced in the expression of some LMG proteins. It is concluded that in transformed cells the regulatory control over the coupling of matrix biosyntheses is impaired.

Standardization of ELISA for detection of antibodies to orthopoxviruses.

The experience with ELISA technique utilized at serological screening for orthopoxviruses in the Republic of Ivory Coast revealed factors reducing the sensitivity and the specificity of the test. It was found out that routine controls such as "positive" and "negative" sera as well as the accepted reading of the results by two-fold or higher increase of OD values as compared to the "negative" control may be not sufficient. A significantly enhanced sensitivity and specificity of reaction was achieved by simultaneous examination of each serum under study with a control antigen. Selection of optimal dilutions of each test component followed by spectrophotometric assay and calculation of results according to the given formula contributed to the same aim. As a result of these improvements the rate of antibody detection among revaccinees was enhanced from 19 to 78.8% and the titres of ELISA and virus neutralization tests correlated in 88% of cases.

[Effect of levamisole on the course of chronic cytomegalovirus infection in women]. / Vliianie levamizola na techenie khronicheskoi tsitomegalovirusnoi infektsii u zhenshchin.

The therapeutic effect of orally given levamisole in the treatment of women with chronic cytomegalovirus infection was studied. The patients were examined serologically, cytologically, virologically; the factors of cell-mediated immunity were also studied. Oral therapy with levamisole resulted in a definite increase of the cell-mediated immunity with a trend for decrease in antibody titres and reduced virus excretion. Evaluation of the clinical effect of levamisole therapy, however, revealed no distinct difference in the outcomes of pregnancy in the groups of women treated or not treated with levamisole.

[Alkaline phosphatase activity of normal and transformed cultured human cells]. / Aktivnost' shchelochnoi fosfatazy v kul'tiviruemykh normal'nykh i transformirovannykh kletkakh cheloveka.

A study was made of the relationship between the activity of alkaline phosphatase and the proliferation of cultured human cells with different replicative potentials. It is shown that alkaline phosphatase plays a role as one of endogenic stimulators of cellular proliferation. The ageing of diploid cells is accompanied by a decrease in the enzyme activity. Maximum activity was observed during a period of logarithmic cell growth. Addition of placental alkaline phosphatase to the synchronized diploid cells stimulated DNA synthesis in the S-phase of the cell cycle. Heteroploid cells with a high growth rate possessed a 30-100 times higher alkaline phosphatase activity than in the diploid cells. Under certain conditions alkaline phosphatase may presumably function as a proteinkinase.

[Selective accumulation of alkaline phosphatase in the microsomes of human cells cultured in the G0-phase of the cell cycle]. / Izbiratel'noe nakoplenie shchelochnoi fosfatazy v mikrosomakh kul'tiviruemykh kletok cheloveka v G0-faze kletochnogo tsikla.

The subcellular distribution of alkaline phosphatase in normal and transformed cultivated human cells was investigated. The total activity of alkaline phosphatase in the transformed cells exceeds that in normal cells 10- (or more) fold and is sharply increased with a rise in the cell population density. The proliferating cells have the same specific activity of alkaline phosphatase in the mitochondrial-lysosomal and microsomal fractions. The transition of normal cells to the G0-phase of the cell cycle is accompanied by an increase in the total enzyme activity and selective accumulation of alkaline phosphatase in the microsomes. An addition of exogenous alkaline phosphatase to intact cells leads to the stimulation of DNA synthesis and to changes in cell morphology. The data obtained suggest that alkaline phosphatase can participate in the destabilization of the cytoskeleton and stimulate cell proliferation. However, in normal cells the selective accumulation of the enzyme in the membranes may hamper its biosynthesis. the activity of alkaline phosphatase thus does not reach the level sufficient for unrestricted cell growth.

[Correlation between the indices of immunity and virus elimination in cytomegalovirus infection]. / Izuchenie korreliatsii mezhdu nekotorymi pokazateliami immuniteta i éliminatsiei virusa pri tsitomegalovirusnoi infektsii.

Correlations between titres of complement-fixing antibody, levels of E-RFC, Ea-RFC, EAC-RFC, EA-RFC, and the intensity of virus excretion in the saliva and urine of women with chronic form of cytomegaly, as well as the effect of decaris in vitro on the level of E-RFC were studied. The highest antibody titres were found in subjects with generalized infection (virus excretion in the saliva and urine) and in cases where high concentrations of the virus were found in the salivary gland cells. In these subjects, total levels of E-RFC and their active subpopulations were decreased. The lowest antibody titres were found in women excreting no virus in the study period. Relative amounts of E-RFC and Ea-RFC in them were within normal ranges. No correlation between the levels of EAC-RFC, EA-RFC, and virus excretion was found. Decaris in vitro normalized the reduced values of E-RFC. Theoretical substantiation of the possibility of using this drug for treatment of patients with cytomegalovirus infection is presented.

[Mechanism of post-synthetic changes in glucose-6-phosphate dehydrogenase in human cells cultured in vitro]. / Mekhanizm postsinteticheskoi modifikatsii gliukozo-6- fosfatdegidrogenazy v kletkakh cheloveka, kul'tiviruemykh in vitro.

The reasons for the decreased stability of glucose-6-phosphate dehydrogenase in transformed human cells were investigated. The enzyme stability was found to be dependent on its subunit composition; the dimeric form possessed a lower stability in comparison with the tetrameric one. An addition of NADP to cell extracts which had partly lost their glucose-6-phosphate dehydrogenase activity, resulted in reactivation and stabilization of the enzyme. The constants for a forward (k1) and back (k2) reactions during stabilization are equal to 2.87 X 10(-3) and 5.77 X 10(-1) s-1, respectively. The inactivation and reactivation kinetics suggest that the enzyme destabilization may also occur inside the cells. The cells contain more than 40% of glucose-6-phosphate dehydrogenase molecules in an inactive form. A mechanism of destabilization and inactivation of glucose-6-phosphate dehydrogenase is proposed, which consists in NADP hydrolysis and enzyme decomposition to inactive monomers which are less stable to proteolysis.

[Role of alkaline phosphatase in the regulation of the stability of glucose-6-phosphate dehydrogenase from human cells cultivated in vitro]. / Rol' shchelochnoi fosfatazy v reguliatsii stabil'nosti gliukozo-6- fosfatdegidrogenazy kletok cheloveka kul'tiviruemykh in vitro.

The causes for different stability of glucose-6-phosphate dehydrogenase in two heteroploid cell strains and in the diploid cell strain of human embryo lungs were investigated. The thermostability of glucose-6-phosphate dehydrogenase was shown to be dependent on the coenzyme (NADP) concentration and to be coupled with the activity of alkaline phosphatase. In diploid and heteroploid cell extracts possessing a low alkaline phosphatase activity glucose-6-phosphate dehydrogenase reveals a high stability. In heteroploid cell extracts having a high activity of alkaline phosphatase a fast hydrolysis of NADP and a decrease of glucose-6-phosphate dehydrogenase stability are observed. Inhibition of alkaline phosphatase by levamisole prior to cell disruption does not increase the stability of glucose-6-phosphate dehydrogenase. Presumably destabilization of glucose-6-phosphate dehydrogenase mediated by alkaline phosphatase occurs in intact cells and is an essential mechanism controlling the enzyme activity.