Evaluation of Collagenase Activity from Crab Hepatopancreas in Different Model Systems.

The effect of Kamchatka crab hepatopancreas containing three collagenolytic isoenzymes Collagenase KK and proteinases of Streptomyces lavendulae on metabolic activity and cell death were carried out on in vitro models. It was shown that changes in the protein structure under the influence of Collagenase KK occurred earlier than under the effect of bacterial proteinases. At the same time, activity of Collagenase KK was significantly higher than that of bacterial proteinases (p<0.01). Both preparations had a pronounced time- and dose-dependent effects on metabolic activity of cells. Collagenase KK had low cytotoxic effect, and cells mainly died by apoptosis. Thus, hepatopancreas collagenase has a high activity and proapoptotic effect on cells and can be used in low concentrations for enzymatic disaggregation of tissues.

[Markers of redox potential of blood leukocytes in acute coronary syndrome, depending on the presence of type 2 diabetes mellitus].

Aim      Evaluating the redox potential of white blood cells (WBC) in acute coronary syndrome (ACS) depending on the presence or absence of type 2 diabetes mellitus (DM2).Material and methods  The study included 100 men and women aged 35 to 65 years who were managed for ACS at the Primary Vascular Department (PVD) of the Vladivostok Clinical Hospital #1. The control group consisted of 30 healthy volunteers matched with ACS patients in major anthropometric characteristics. Examinations were performed according to clinical recommendations. Blood was withdrawn for measuring cell activity of enzymes (superoxide dismutase, SOD; succinate dehydrogenase, SDH; and glutathione reductase, GR) and serum concentration of malonic dialdehyde (MDA). Based on the ACS type, all patients were divided into 3 main ACS groups, and then the groups were subdivided into subgroups based on the presence of DM2.Results Development of ACS was associated with changes in WBC redox potential. These changes were characterized by a significant decrease in SDH activity in all ACS patients, irrespective of their ACS type, and a moderate decease in GR in patients with myocardial infarction compared to patients with unstable angina and healthy volunteers. At the same time, the SOD activity and MDA concentration were practically unchanged compared to the control group. There were practically no significant differences in the enzyme activities between the ACS subgroups with or without DM2.Conclusion      The WBC activities of SDH and GR on day 1 of ACS can be considered as the indicators for early diagnosis of mitochondrial dysfunction resulting from the cardiovascular catastrophe as well as the markers for impaired primary cell defense. MDA and SOD values are not informative for determining the intensity of oxidative stress and further damage of the antioxidant system.

Antihistamines Modulate Functional Activity of Macrophages.

We compared the effects of the first-, second- and third-generation antihistamines in different doses on enzyme activity and cytokine production by macrophages and their death using an in vitro model. It was found that decreasing the dose led to an increase in the number of viable cells; after contact with second-generation antihistamines (loratadine, desloratadine), apoptosis of macrophages predominated. A dose-dependent increase in activity of ATPase and 5'-AMP with less pronounced effect of second-generation drugs was revealed. It was shown that under the influence of drugs, macrophages do not produce IL-1ß, but actively synthesize TNFα and IL-10, which indicates the immunomodulatory properties of these drugs.

Effect of Mild Traumatic Brain Injury on Behavioral Reactions and Neocortical Morphology in Rats.

The original weight-drop model was employed to examine the effect of mild traumatic brain injury (TBI) on behavioral phenotype and neocortical morphology in rats. The neurological examination of rats with moderate TBI revealed the focal symptoms corresponding to pronounced neurological disorders, whereas in rats after mild TBI, there were only minor coordination disorders. On day 7 after injury, the rats with mild TBI demonstrated enhanced anxiety assessed by conditioned passive avoidance response. The morphometric analysis of the brain tissues revealed narrowing of the capillaries and increased score of hyperchromic neocortical neurons, which attested to cerebral hypoxia. The manifestations of mild TBI in original rat model demonstrated a close similarity to the symptoms of TBI in humans.

[Aortic stiffness and content of adipokines in the serum in persons of European and South Asian ethnic].

AIM: To evaluate the cardiovascular risk (CVR) based on arterial stiffness and content of adipokines in young-aged persons of different ethnicity (European and South Asian). MATERIALS AND METHODS: 290 persons of European (Slavic) and South Asian (Korean) ethnicity aged from 19 to 49 years with and without arterial hypertension (AH) were examined. Clinical, anthropometric, laboratory examinations were performed, levels of resistin and adiponectin of blood were assessed. Total CVR was assessed by SCORE scale, patients under the age of 40 years were assessed by relative risk scale. Aortic stiffness was examined by non-invasive arteriography. RESULTS: Patients of European ethnicity had higher blood pressure (BP), body mass index (BMI), waist circumference (WC), levels of resistin and adiponectin. Pulse wave velocity in the aorta (PWVA) did not differ significantly in ethnic groups. According to the SCORE scale in individuals of the European and South Asian races in general groups and groups with arterial hypertension a moderate absolute risk was determined, in individuals under 40 years of age a moderate relative risk was determined without a significant difference between the groups. However increased levels of PWVA (more than 10 m/s) were registered more often in Korean ethnicity (46.9% compared to Slavic ethnicity, 22.2%). Closer reliable correlations between the level of BP and BMI, WC, PWVA were revealed in Korean ethnicity. Ethnic differences in correlation of adipokines in blood and their dependence on anthropometric and hemodynamic characteristics were described. CONCLUSION: The assessment of CVR according to traditional scales does not always accurately represent its real level. New information was obtained on the features of adipokine metabolism and its connections with early manifestations of vascular remodeling in young-aged depending on the race. Taking into account ethnic differences, we recommend in-depth diagnostics of CVR in South Asians. The data can be useful for the design of personalized programs for the diagnostics and assessment of CVR.

Functional State of Mesenchymal Stem Cells upon Exposure to Bioactive Coatings on Titanium Alloys.

Bioactive coatings on implants affect osteogenic differentiation of mesenchymal stem cells (MSC). We studied the morphofunctional state of bone marrow MSC cultured on the surface of calcium phosphate coatings on titanium formed by plasma electrolytic oxidation (PEO). The biocompatible properties of the coatings manifested in the absence of the cytotoxic effect on cells. High expression of receptors (CD90, CD29, and CD106), enhanced synthesis of osteocalcin and osteopontin, and changes in surface architectonics of MSC adherent to the samples confirmed osteoinductive properties of the calcium phosphate PEO coating.

[Morphofunctional features of the cerebral cortex in chronic obstructive pulmonary disease concurrent with ischemic stroke]. / Morfofunktsional'nye osobennosti kory bol'shikh polusharii mozga pri sochetanii khronicheskoi obstruktivnoi bolezni legkikh i ishemicheskogo insul'ta.

OBJECTIVE: To determine the structural changes in the cerebral cortex tissues, which are characteristic of the severe course of chronic obstructive pulmonary disease (COPD) and ischemic cerebral stroke (ICS). MATERIAL AND METHODS: The autopsy material of the microcirculatory bed of the cerebral cortex from 18 people aged 48-64 years who had died from COPD, ICS, and a concurrence of these conditions underwent histological examination and morphometric analysis. RESULTS: The state of the brain from the persons who died from ICS was characterized by a decline in the total number of neurons and glial elements, by structural and regional blood flow changes with obvious venous hyperemia, stasis, and red blood cell aggregation in the vessels. There was pericellular and perivascular edema in the perifocal area of the brain from people who died from COPD, as well as morphological signs of cerebral hemodynamic disorders. The brain structural features in ICS concurrent with COPD included a double decrease in the numerical density of capillaries, their hyperemia, perivascular edema, and venous plethora, a substantial change in the diameter of non-muscular venules, parenchymal atrophy, and stromal sclerosis, indicating the presence of a chronic process. CONCLUSION: Patients with ICS in the presence of COPD versus those who died from the isolated variants of these conditions were recorded to have an obvious decrease in the numerical density of capillaries, their hyperemia, and a pronounced change in the structure of the cerebral vein wall, which is a consequence of multisystem processes associated with COPD.

Behavioral and Cognitive Reactions of Rats with Experimental Chronic Obstructive Lung Diseases of Different Degree.

The study presents the results of behavioral phenotyping of rats with experimental chronic obstructive pulmonary disease (eCOPD) of different severity modeled by using a combination of inductors (purified papain and bacterial LPS) in increasing concentrations. Locomotor and exploratory activity, spatial memory, and anxiety were evaluated. In mild eCOPD, locomotor and exploratory activity increased, while other parameters did not differ from the control values. Rats with moderate eCOPD demonstrated moderately restricted locomotor function and increased anxiety. Severe eCOPD was associated with pronounced changes in all analyzed parameters. Correlation analysis revealed relationships between some behavioral parameters and blood oxygen saturation, inspiratory lung volume, and respiration rate. This indicates the role of respiratory failure in the pathogenesis of cognitive disorders.

Radioprotective Properties of Neomitilan in Experimental Radiation Pneumonia.

The efficiency of neomitilan, a polysaccharide isolated from Crenomytilus grayanus mussels, was studied in experimental model of radiation pneumonia (irradiation of the animal lungs with a total dose of 14 and 28 Gy). Histological study showed that an increase in the number of nucleated components in the lungs of animals subjected to neomitilan inhalation prior to irradiation. Proliferation of bronchial epitheliocytes and the formation of additional lymphoid structures were also revealed in these animals. The absence of a tendency to lung tissue recovery in animals irradiated with 28 Gy is due to extreme radiation toxicity.

THE STRUCTURAL CHANGES OF MACROPHAGES INFECTED WITH TICK-BORNE ENCEPHALITIS VIRUS.

Macrophages belong to the innate immune cells and play a key role in the pathogenesis of viral infections. The results of ultrastructural study of macrophages infected with tick-borne encephalitis virus (TBEV), the Flavivirus family, pathogens of human infections, affecting the nervous system, were presented. With the assistance of virological methods was found that the TBEV are absorbed by macrophages and replication in them. An ultrastructural study has shown that the virus enters into the cytoplasm by local destruction of plasmalemma and newly synthesized virus particles exited from the cell by same. Simultaneously there is a seal of perinuclear cytoplasm space, where found in a large number of ribosomes, microfilaments, ribonucleoprotein fibers and viral special structure: nucleocapsids, tubular formations and viral layers (fabrics). On the surface of last structures the newly synthesized virus particles were visualized. Thus, the evidence shows that macrophages play a role in the spread of TBEV, being for their the target cell. As active antigen presenting cells the macrophages can modulate the protective response of the body and influence on the pathogenesis of tick-borne encephalitis.

Morphogenesis of Experimental Infection Caused by Plasmid Variants of Yersinia pseudotuberculosis.

The dynamics of pathomorphological changes in response to infection with plasmid variants of Yersinia pseudotuberculosis was studied in experimental animals. Variability of cell injuries in pseudotuberculosis histopathology depended on the plasmid-associated virulence of the infection agent. Infection with highly virulent two-plasmid strain pYV48:pVM82 MDa and Y. pseudotuberculosis strain with low virulence with the only plasmid pVM82 MDa led to the development of cell destruction (necrosis and apoptosis) in the target organs. Apoptosis predominated in response to infection by plasmid variant pVM82 MDa with low virulence.

[Pathomorphism of Hantavirus infection under prolonged heat stress]. / Patomorfoz khantavirusnoi infektsii na fone prolongirovannogo teplovogo stressa.

AIM: to provide the morphological characteristics of experimental Hantavirus infection under heat stress conditions to identify the possibilities of its modeling in resistant laboratory animals. MATERIAL AND METHODS: Experiments were carried out on outbred albino mice that were divided into 4 groups: 1) intact mice unexposed to heating; 2) those exposed to heating; 3) Hantavirus-infected animals unexposed to preheating; 4) those exposed to preheating. The animals in Groups 2-4 were long exposed to heat stress at a temperature of 30 °C for 3 hours during 3 days. Strain Aa 60343 (PM-79-95) of the Far East genovariant of Hantaan virus (the genus Hantavirus, family Bunyaviridae) from the collection of the G.P. Somov Research Institute of Epidemiology and Microbiology was used to induce infection. The animals in Groups 3 and 4 were intraperitoneally injected with 700 FFU of Hantavirus per mouse. Materials (lung, liver, kidney, and spleen) taken from Groups 2-4 animals were collected for histological examination on days 1, 3, 7 and 14 of observation. RESULTS: The intact albino mice in Group 1 showed no histopathological changes in the organs. After heat exposure, Group 2 animals were found to have an immunomorphological response in the interstitial tissues of the lung, liver and kidney in partial lymphoid hypoplasia of the spleen. There were no signs of inapparent infection in the presence of marked immunomorphological changes in the organs in Group 3 of hantavirus-infected animals unexposed to preheating. In Group 4, those exposed to preheating exhibited dystrophic and destructive changes in the target organs (lungs, kidneys) in the presence of immunodeficiency manifestations of manifestations that were more pronounced in dead animals. CONCLUSION: In an unresponsive model (adult albino mice), Hantavirus infection caused only obvious immunomorphological changes in the organs. Prolonged preheat stress in the hantavirus-infected animals promoted inapparent infection and morphological manifestations of induced secondary immunodeficiency that was responsible for the manifestation of an infectious process in some animals.

[PLASMID-ASSOCIATED VIRULENCE OF YERSINIA PSEUDOTUBERCULOSIS AND INFECTIOUS PROCESS].

Literature data regarding genetically-determined pathogenicity factors of Y pseudotuberculo- sis and associated manifestations of this infection caused by various plasmid types of the causative agent are generalized. Principal attention is given to features of cell-tissue alterations mediated by virulence plasmid pYV, as well as effects of pathogenicity of an understudied pVM82 plasmid present only in Y pseudotuberculosis sttains causing clinical-epidemic manifestation of the infec- tions as Far East scarlet-like fever (FESLF). The data obtained on the ability of far-eastern strains to produceYPMa super-antigenj Ypseudotuberculosis-derivative mitogenA, probablygive evidence on its key role in FESLF pathogenesis. Variability of damage of innate immunity cells and target- organs caused by various plasmid types of Y pseudotuberculosis by virulence could determine polymorphism of clinical-morphological manifestations of this infection. In-depth understanding of dependency of immune pathogenesis mechanisms of the disease on molecular characteristics of the causative agent opens up-perspectives of enhancement of diagnostics and prognosis of the severity of the course of pseudotuberculosis and yersiniosis in human in general.

[The metabolism of the innate immunity cells in bacterial infections].

Metabolic activity of innate immunity cells infected by various doses of Gram-negative (Yersinia pseudotuberculosis, Salmonella enteritidis) and Gram-positive (Staphylococcus aureus, Listeria monocytogenes) bacteria has been investigated. Using various animal models we found that during the initial period (up to 2 days) changes of infection in cellular responses depend on the type of the pathogen. In response to infection caused by Gram-negative bacteria predominant neutrophil accumulation in the foci of inflammation was observed, while Gram-positive bacteria induced preferential accumulation of macrophages. The study of metabolism of these cells showed that the response of terminally differentiated primed phagocytes to pathogen appearance was higher than in cells circulating in blood. In addition to the priming state the phagocyte reactivity is influenced by the bacterial load. At a low phagocyte/microbe ratio the cells reaction is almost undetectable, while an excess of microorganisms causes (despite of the increase of the phagocytic parameters) the hyperactivation of cell metabolism and production of maximal amounts of bactericide agents, which exhibit a damaging effect on the cell itself.

Effects of immunomodulators on functional activity of innate immunity cells infected with Streptococcus pneumoniae.

Low activity of bactericidal enzymes was found in innate immunity cells infected with S. pneumonia. The death of these cells was fastened under these conditions. On the contrary, treatment with antibiotic maxifloxacin was followed by an increase in activity of bactericidal enzymes in phagocytes and induced their death via necrosis. Analysis of the therapeutic properties of immunomodulators tinrostim and licopid in combination with maxifloxacin showed that these combinations correct functional activity of cells infected with S. pneumonia.

Effect of thermolabile toxin from Yersinia pseudotuberculosis on functions of innate immunity cells.

The thermolabile toxin of Yersinia pseudotuberculosis produces a selective dose-dependent stimulating effect on functional activity of innate immunity cells. Prolonged apoptosis-inducing action of the toxin was associated with activation of enzymes of the oxygen-dependent system (LDH and myeloperoxidase) at the early terms of observation (up to 3 h). In turn, increased number of macrophages with apoptotic changes was noted at the early stages of contact with the thermolabile toxin (5 h), and its further growth was observed against the background of activation of mitochondrial enzymes and production of NO metabolites.

Morphological validation of hydroxyethylstarch use during the acute period of severe brain injury.

The effects of 10% hydroxyethylstarch solution on the cerebral microcirculatory bed and the blood-brain barrier were studied in Wistar rats during the acute period of severe brain injury. Positive changes in the morphometric values of the cerebral capillaries were observed in animals receiving intravenous injections of the drug, which promoted reduction of vascular permeability and cerebral tissue edema. These results confirmed the efficiency of hydroxyethylstarch as a component of infusion therapy in patients with severe brain injury.

Pathomorphosis of experimental infection in mice, infected by Streptococcus pneumoniae, under the effect of immunotropic drugs.

Pathomorphological changes in the organs of animals intranasally infected with Streptococcus pneumoniae were studied under conditions of immunotropic therapy added to antibiotic therapy. The pathomorphosis in the lungs, spleen, and thymus in animals treated with likopid, tinrostim, and roncoleukin was described. A positive time course of the pathological process in experimental animals in comparison with intact animals and animals receiving no immunotropic drugs was demonstrated.

[The physiological role of nitric oxide in infectious process].

This review presents the recent data on the physiological role of the molecule nitric oxide (NO) and its derivatives in the pathogenesis of bacterial and viral infection. Indicated that the effect of NO on individual cells involved in inflammation and immune regulation may be ambiguous, and depends on the cellular environment on the concentration NO and other factors. Thus, the interaction between reactive oxygen and NO derivatives provides a molecular basis for synergy between the respiratory burst and synthesis of NO, which leads to the formation of peroxynitrite, which has powerful bactericidal potential. At the same time production of NO can counteract the oxidant damage to cell membranes by blocking the oxidation of lipids and protect cells from damage, for example, by inhibiting the respiratory burst. Thus, nitrate and oxidative stress observed in pathophysiological states, has a modulating effect on the function of cells and tissues of various organs.

[The new views to pseudotuberculosis pathology].

For the first time the nature of cell damages under pseudotuberculosis as a generalized infection has been analyzed by positions of the modern knowledge about types of the cell death. The own and literature data have pointed to presence the apoptosis-induced effect of Yersinia pseudotuberculosis, a causative agent of this infection. In conclusion, the typical pathological changes for pseudotuberculosis infection such as granuloma formation with the central karyorrhexis could be appearance of apoptotic and secondary necrotic changes.