Local and Systemic Functional Responses of Mouse Macrophages to Intravaginal Infection with Type 2 Herpes Simplex Virus and Vaccination.

Activity of cathepsin D and phagocytosis of macrophages from vaginal lavage fluid, peritoneal exudation, and spleen were studied in mice of sensitive (DBA/2) and resistant (BALB/c) lines after intravaginal infection with type 2 herpes simplex virus and vaccination. Activity of cathepsin D and intensity of phagocytosis (irrespective of the macrophage source) and their ratio in BALB/c mice in early terms after infection were close to the control levels taken as a unit. In DBA/2 mice, these parameters and their balance were shifted and changes in cathepsin D activity depended on the time after challenge. Activities of cellular and extracellular cathepsin D increased sharply on day 1 postinfection under conditions of local virus interaction with the vaginal mucosa and activation of the pathological process. Later, after generalization of the infection, activity of cathepsin D decreased, while phagocytosis increased in all the studied macrophage populations. Vaccination corrected the cathepsin D/phagocytosis imbalance and created conditions for rapid elimination of the virus.

[Dual role of photosensibilizator merocyanine 540 as a bactericidal agent and immune reaction regulator].

AIM: Develop conditions for inactivation of staphylococcus by using photosensibilizator merocyanine 540 (MC540) for the production of antigenic preparation (AP). Study some of immune reactions to AP and the possibility of regulation of DTH reaction to AP under the effect of MC540. MATERIALS AND METHODS: Merocyanine 540 (MC540, Sigma-Aldrich, Switzerland) is used in the study. MC540 and Staphylococcus aureus, strain 78 (Sa78) were irradiated by light of a mercury-quartz lamp DRSH-250 (Zelenograd). C56BL/6 line mice were immunized once by subcutaneous administration of AP. DTH reaction was tested 7 days after the immunization. Functional activity of peritoneal exudate macrophages was determined 1 and 9 days after the immunization. Immune modulating effect of MC540 in DTH was determined after its per os administration to mice 1 hour after AP sensibilization. RESULTS: In order to obtain AP, S. aureus suspension at the concentration of 2.5 x 10(7) CFU/ml in 25 microM MC540 solution and 0.25 M NaCl solution were exposed to irradiation for 5 minutes. During DTH reaction induction its intensity dependence on AP dose was revealed. A persistent increase of a lysosomatic enzyme cathepsin D in macrophages of peritoneal exudate after a single administration of AP was noted. During MC540 irradiation an accumulation of photoproducts that have a pronounced immune suppression effect in DTH reaction had a dose-dependent character. CONCLUSION: Use of saline allows to increase bactericidal potential of a photosensibilizator (PS). However during therapy of localized forms of infection a possible immune modulating effect of PS on macro organism should be considered. By varying PS dose and irradiation time not only maximum bactericidal effect can be achieved but also regulation of inflammatory reactions in the area of PS effect can be ensured.

[The features of system organization of the brain activity during different sleep stages and transition states].

Electropoligraphical study of the natural night sleep in 16 adults with the use of correlation, coherent, cluster and factor analysis were used to obtain new data describing the active nature of sleep, which is expressed especially in periods of falling asleep and the transition from one stage to another. It is shown that the process of falling asleep and deeper sleep is accompanied by intense reorganization of cortico-subcortical relations, which is reflected in the dynamics ofcrosscorrelation and coherent estimates of interrelations of biopotentials of the brain. The results of factor analysis of multichannel EEG heterogeneity of the transition process from wakefulness to sleep is manifested in significant changes of I, II and III factors weight during I(B) stage of sleep, which may reflect changes in the degree of contribution of the main integrative brain systems in the reorganization of its integral activity. A considerable increase in the I factor weight (reflecting the generalized modulatory brainstem effect on the cortex), along with a decrease in the balance of factors II and III (associated with organization of fronto-occipital and interhemispheric interactions) clearly indicates a special role of sleep synchronizing influences from the brain stem in the development of this initial stage. Reduction of EEG interhemispheric interrelations in the anterior and inferior frontal areas with the deepening of sleep may be indication of the reorganization of the frontal areas activity associated with the coordinated increasing of inactivation process in the cortex of both hemispheres. Degree of stability of the spatial structure of interregional interactions of different brain cortex areas (according to the analysis of average dispersion of crosscorrelation EEG relations) increases on falling asleep with the onset of stage I(A), but with the transition to the stage I(B) there is a significant increase of instability of values EEG crosscorrelation. With the deepening of sleep the subsequent decrease of the dispersion of EEG crosscorrelations in frontal cortex is revealed. During REM sleep the dispersion levels of inter-regional interactions increases as much as possible, especially for EEG crosscorrelations of posterotemporal and inferiofrontal parts of both hemispheres.

[Organisation of the hemisphere interaction in preschool children and in adults during mental speech production and speech perception].

The main goal of this study was to investigate reorganisation of the EEG systemic interactions spatial structure during mental speech production in preschool children: generating sentences from the set of words and generating words from the set of phonemes. In both cases, interhemisphere biopotential relations significantly increased as compared with the baseline (resting with closed eyes). Results of the EEG cross-correlation and coherent analyses showed that during verbal tasks marked intensification of hemisphere interaction was observed. High coefficients of statistic similarity between intercortical interactions patterns in adults and children during sentence and words generating were observed (SC = 0.71 & 0.62 respectively). Opposite to that, lesser coefficients of statistic similarity were observed between these two groups during grammar and semantic mistakes identification (SC < or = 0.50). According to this data we could expect a relatively high maturation level of central mechanisms which underlie the processes of speech production rather than mechanisms which underlie grammar and semantic mistakes identification in preschool children.

[Features of cortex areas system interactions in the left and right brain hemospheres during different human sleep stages].

The article is devoted to analysis of research brain biopotential field structure during different stages of natural night sleep in adults. The special attention is given to distinctions of the spatial organization of EEG interregional relations in the left and right hemispheres which are shown most strongly in the II and III sleep stages. The special role of frontal cortical areas in reorganization of brain system activity during falling asleep is highlighted. Essential strengthening of hemispherical interaction of cortex biopotentials fluctuations during falling asleep--a stage of I(B)--along with its reduction in central and occipital cortical areas is shown during a slow wave sleep (II--IV stage) versus wakefulness.

[Peculiarities of disorders of brain bioelectrical activity spatial-time organization in patients with different consciousness depression after severe head injury].

Specific changes of bioelectrical brain activity was found in 27 patients with different level of posttraumatic consciousness depression by the methods of crosscorrelation, coherence and factor analysis of EEG. The changes of activity of morphofunctional systems of intracerebral integrations were revealed partially by decreasing of unspecific activity from brainstem structures reflected with increasing of slow wave activity and decreasing of EEG coherence in alpha- and beta-range. Depression of system organization of interconnections of bioelectrical brain activity in frontal and occipital regions of both hemispheres was also detected, and testified about decreasing of intercortical and thalamocortical brain system action under brain dislocation. The changes of integrative brain system activity, provides interhemispheric interaction, had the specific characted. Our results propose a "facilitation" of activity of system, providing "direct" interhemispheric connections through corpus callosum and other commissural tracts of telencephalon as a sequel of mesodiencephalon structures depression with steady reciprocal, antiphase relations of slow weve activity in symmetrical areas of hemispheres in coma II patients. The data of our research had shown no complete disintegration of system brain activity in coma II patients in spite of consciousness and brainstem reflexes depression.

[Ontogenetic formation of stereognosis brain mechanism (EEG study)].

In this study the age specificity of inter- and intrahemispheric relations during stereognostic task (tactile identification of predetermined chess-like pieces with right or left hand) in adults and children of 5-6, 7-8 an 9-10 years old is considered. The combined pattern of cortical interactions was revealed in adults. Its appeared in significant increasing (compared to baseline) of interhemispheric EEG relations along with increasing of system interactions of anterior and posterior cortex areas bioelectrical activity. This combined pattern is observed both in case of using EEG coherence and correlation analysis. In children interhemispheric EEG relations increasing is appeared already in 5-6 years (maximum in 7-8 yo.), whereas increasing of EEG relations in fronto-occipital direction expanses gradually with age, and only in age 9-10 approaches to definitive level, typical for adults. The data obtained evidence the gradually formation in postnatal ontogenesis of central mechanisms of such important function--stereognosis. Our results suppose that brain mechanisms of right (leading) hand stereognostic activity maturate earlier than left (nonleading) hand activity. Heterochronic involving in stereognostic activity of inter- and intrahemispheric relations can be closely related with gradual and heterochronic processes of commissural and associative hemispheric tracts mielinization.

Functional activity of peritoneal macrophages from sensitive and resistant mouse strains during intravaginal infection with herpes simplex virus type 2 and mucosal vaccination.

In the early period after intravaginal infection with herpes simplex virus type 2 (2 h), macrophages from sensitive DBA/2 mice were characterized by higher capacity to engulf the antigen, decreased function of the lysosomal apparatus, lower activity of cathepsin D, and reduced oxygen metabolism compared to cells from resistant BALB/c mice. Mucosal vaccination with herpes vaccine and hyaluronic acid promoted the increase in functional activity of macrophages and improved survival of sensitive mice (by 60%).

Comparative study of macrophage response in mice after DNA immunization and infection with herpes simplex virus type 1.

Functional activity of macrophages and intensity of T cell immune response in mice were studied after intravaginal and intraperitoneal infection with herpes simplex virus type 1 and DNA vaccination in combination with adjuvant treatment (recombinant granulocyte-macrophage colony-stimulating factor and glucosaminylmuramyl dipeptide). DNA vaccination induced a virus-specific T cell immune response with no macrophagic inflammatory reaction. Infection with herpes simplex virus type 1 was accompanied by sustained inflammation, but not by the T cell immune response.

[The correction action of Phosprenyl and Gamavit on the functional activity of mouse peritoneal macrophages in response to high doses of Staphylococcus aureus alpha-toxin].

The study of the functional activity of peritoneal macrophages of BALB/c mice at different stages of the toxic action caused by S. aureus alpha-toxin (ST) was carried out. The analysis of the dynamics of toxic reaction revealed the main critical points of triggering necrotic processes: the first hour and day 2. One hour after the injection of large doses of ST a sharp increase in the process of antigen binding with its subsequent sharp decrease. Simultaneously, a decrease in the activity of the lysosomal enzymes cathepsin D and acidic phosphatase was established, which was indicative of the destabilization of both lysosomal and cellular macrophage membranes. The increase of oxygen metabolism on day 2, together with the release of lysosomal proteases into the extracellular area, correlated with the maximum death rate of mice and served as the main index of the development of necrosis. The prophylactic and therapeutic use of the preparations Gamavit and Phosprenyl revealed their antitoxic activity and capacityfor stimulating the level of natural body resistance.

[Mitochondrial H+-ATPase: identification of subunits of the F0 subcomplex contacting with membrane lipids]. / H+-ATP-asa mitokhondrii: identifikatsiia sub''edinits subkompleksa F0, kontaktiruiushchikh s lipidami membrany.

The subunits of the F0 membrane sector of bovine heart mitochondrial H(+)-ATPase that contact the lipids of the mitochondrial inner membrane were identified with the use of specially synthesized proteoliposomes that contained active mitochondrial H(+)-ATPase and a photoreactive lipid, which was 1-acyl-2-[12-(diazocyclopentadiene-2-carbonylamino)-[12-14C]dodecanoyl]-sn- glycero-3-phosphocholine, 1-acyl-2-[11-([125I]diazoiodocyclopentadiene-2-carbonyloxy)undecanoyl]-sn- glycero-3-phosphocholine, or 1-acyl-2-[12-(diazocyclopentadiene-2-carbonylamino)dodecanoyl]-sn-glycero- 3-phosphocholine, where acyl is a mixture of the residues of palmitic (70%) and stearic (30%) acids. An analysis of the cross-linked products obtained upon the UV-irradiation of these proteoliposomes indicated that subunits c and a of the F0 membrane sector contact the lipids. The cross-linked products were identified by SDS-PAGE and MALDI mass spectrometry.

[Protein import into mitochondria]. / Import belkov v mitokhondrii.

This review is focused on the import of processable precursor proteins into the mitochondrial matrix; the import of carrier proteins into the inner mitochondrial membrane is also briefly discussed. Post- and cotranslational theories of the import, specific features of the presequence structures, and effects of some cytosolic factors on the import of precursor proteins are reviewed. The data on the structure of the protein translocases of the outer (TOM complex) and the inner (TIM complex) membranes of mitochondria and the current models of the precursor protein import by these translocases are also summarized.

[Protein composition of venoms from several species of tropical ants and their effect on mitochondrial H+-ATPase]. / Issledovanie belkovogo sostava iadov nekotorykh vidov tropicheskikh murav'ev i ikh vliianiia na H+-ATPazu mitokhondrii.

Protein compositions of venoms of South-American stinging ants, Ectatomma tuberculatum, Paraponera clavata (subfamily Ponerinae), and "tangarana" were analyzed. The venom of E. tuberculatum displayed the most complex protein composition (more than 15 polypeptides). The water-soluble fraction of the venoms of P. clavata and "tangarana" contained acidic proteins (pI < 3.5 to 5.2), whereas the venom of E. tuberculatum contained predominantly basic proteins (pI 8 to > 9.5). N-Terminal residues and N-terminal sequences of a number of polypeptides were determined. High-molecular-mass polypeptides of the P. clavata venom slightly stimulated the ATPase activity of mitochondrial F1-ATPase. Low-molecular-mass nonprotein components of this venom significantly inhibited the ATPase activity of submitochondrial particles and F1-ATPase from bovine heart mitochondria. The venoms of E. tuberculatum and "tangarana" produced no effect on the ATPase activity.

[The Amben regulation of the intestinal microflora and macrophage functional activity in an experiment]. / Reguliatsiia ambenom kishechnoi mikroflory i funktsional'noi aktivnosti makrofagov v éksperimente.

The possibility of the correction of intestinal microflora disorders and the functional activity of macrophages in dysbiosis, caused by the intragastric administration of ampiox, with the use of amben (PAMBA), an inhibitor of proteolytic enzymes, was studied. Quantitative and qualitative changes in the main representatives of automicroflora, the functional activity of macrophages in the phagocytosis of 51Cr-labeled sheep red blood cells, the intensity of protein synthesis, the content of cathepsin D, acidic phosphatase and nitro blue tetrazolium activity were determined. The combined administration of ampiox and amben normalized quantitative and qualitative ratios of the main representatives of intestinal microflora, as well as the characteristics of macrophage functional activity, studied in this investigation. The administration of amben to intact animals was found to stimulate bifido- and lactoflora.

[The mechanism of the combined action of immunomodulators on phagocytic cells]. / K mekhanizmu kombinirovannogo vozdeistviia immunomoduliatorov na fagotsitarnye kletki.

The study revealed that the injection of different immunomodulators (salmosan, levamisole, BCG, prodigiosan) or such irritants as meat-peptone broth led to the formation of heterogeneous macrophage populations, differing in their phagocytic function, oxidation metabolism, secretion of protein products. The additional injection of immunomodulators led neither to further increase in the influx of cells into the abdominal cavity, nor to further activation of the phagocytic function in all cases, irrespective of background and additional preparations used in the experiment. The response was determined by the first stimulus. Each of these macrophage populations was seemingly in a characteristic state of self maintenance during a certain period when it was insensitive to the second stimulus, i.e. the existence of a certain refractory period was revealed. The effect produced by the additional injection was registered only at the expiration of the refractory period. Therefore, the data on the presence of a certain refractory period and its duration for each immunomodulator should be established and taken into consideration in working out the schedule of their combined use.

[Effect of regional (wound) revaccination with tetanus anatoxin on the functional activity of immunocompetent cells in the region]. / Vliianie regionarnoi (ranevoi) revaktsinatsii stolbniachnym anatoksinom na funktsional'nuiu aktivnost' immunokompetentnykh kletok regiona.

The study made with the use of complex methods established that the local (wound) application of tetanus toxoid rapidly made the manifestation of the lysosomal apparatus more pronounced, increased the oxidizing activity (determined in the nitro blue tetrazolium test) and phagocytic activity of the mononuclear phagocytizing system in the wound and in the regional lymph nodes. The wound application of tetanus toxoid significantly increased blast transformation of T lymphocytes in guinea pigs simultaneously with tetanus wound infection. The study confirmed the pathogenetic expediency of the proposed method for the stimulation of anti-tetanus immunity by the application of tetanus toxoid on the wound which specifically inhibited the primary stage of the infectious process.