Guinea Pig Blood Research Articles

The effect of thiocetam on the parameters of the nitric oxide system under the conditions of the experimental periodontitis and immobilization stress formation

Introduction and aim. The aim of this work is to study the parameters of the nitric oxide (NO) system in the blood of guinea pigs under the conditions of the experimental periodontitis (EP) and immobilization stress (IS) formation and to evaluate the effectiveness of thiocetam use. Material and methods. Experimental studies were performed on 50 guinea pigs (males, body weight 0.18–0.21 kg) which were divided into five groups (10 in each): the first group were intact animals as control; the second experimental group were animals with experimental periodontitis under conditions of immobilization stress (3rd day), the third group included guinea pigs with EP and IS on the 5th day of the combined model process, group IV – animals with EP and IS 15th day (without administration of thiocetam) and group V – animals on the 15th day of experiment with EP and IS after use of thiocetam. Results. As a result of this research, changes in the activity of the NO system in the blood were observed, namely an increase in the level of stable metabolites and an increase in the activity of total NO-synthase, which is accompanied by a compensatory inhibition of the L-arginine activity, and these indicators were most pronounced in the late stages of EP and IS formation. Conclusion. The use of thiocetam showed a corrective effect on the changed variables of NO metabolism in the peripheral blood of guinea pigs under the conditions of the EP and IS development.

Assimilation of feed nutrients by Pietren breed pigs depending on their sex

Each breed and type of pig is characterized by its unique complex of biological properties. In addition, pigs of different sexes realize their genetic potential to a different extent. Therefore, research aimed at optimizing the feeding rations of different sex-age groups in pigs of meat genotypes, namely the Pietren breed, depending on their sex, is relevant. Objective. To determine the efficiency of assimilation offeed nutrients by pigs of the Pietren breed, depending on their gender. Methods. The research was conducted in the conditions of the physiological yard of the Institute of Pig Breeding and AIP of the NAAS on 12 pigs with an average weight of 65 kg, namely 6 boars and 6 gilts, which were divided into two groups. Selected average samples of feed and excrement were studied in the laboratory offeeding, physiology and animal health of Institute of Pig Breeding and AIP NAAS. The biochemical composition of the blood of experimental young pigs was investigated using commercial kits from the company "Filisit Diagnostics" Ukraine. Results. It was established that the coefficients of digestibility of nutrients offeed for Pietren pigs do not significantly depend on their gender. However, there is a tendency for 3.05 % better digestibility of mineral substances in piglets. Piglets have better digestibility o f fiber and crude protein, respectively, by 19.20 % and 2.65 %. The average daily balance of Nitrogen and Magnesium did not differ significantly. As for the average daily balance of Calcium and Phosphorus, the excretion of these elements with feces and urine in boars was somewhat less, and it was retained in the body, accordingly, slightly more, but due to the rather significant variability of the indicators, no probable difference between the groups was detected. It was found that the content of Calcium, probably by 17.87 % (р<0.05), and Phosphorus by 9.7%, was higher in the blood of guinea pigs. The content of total protein in the blood plasma of wild boars was probably (p<0.05) higher by 12.5 %, creatinine was probably (p<0.05) higher by 10.44 %, the activity of aspartate aminotransferase was probably (p<0.05) higher by 10.34 %. Conclusions. It was established that the coefficients of digestibility of nutrients offeedfor Pietren pigs do not significantly depend on their gender. However, the analysis of hematological indicators of blood can indicate a higher level of metabolism in boars. Keywords: guinea pigs, pigs, Pietren breed, digestibility coefficient, nutrients, sex, feed, hematological indicators.

Features of oxygen-independent metabolism of neutrophils in the animals sensitized with non-agglutinogenic Brucella strain

The results of evaluation of the peculiarities of functioning of non-enzymatic systems of neutrophils of peripheral blood of guinea pigs before immunization and on the 7th, 14th, 21st, 28th, 42nd, 55th, 69th, 125th days after immunization with Brucella R-strain are presented. Two groups of agouti guinea pigs were formed for the study: control and experimental. The experimental animals were immunized with a suspension of Brucella abortus strain 16/4 at a dose of 1 billion CFU/ml subcutaneously; the control pigs were intact to brucellosis. The activity of antimicrobial systems was evaluated by determining the indices of cationic proteins by photometric method in spontaneous and induced variants. Experimental samples of brucellosis antigens were used as specific inducers in the reaction procedure. It was found that the use of Brucella disintegrans at a concentration of 50 μg/ml and corpuscular antigens at a concentration of 100 μg/ml by protein in the test with cationic proteins as inducers of disintegrants has a stimulating effect on the intensity of cationic protein production in the cell suspension of leukocytes. Stimulation indices were calculated at all observation periods as the ratio of the index of the stimulated sample to the index of the sample without antigen load. Two peaks of stimulating activity of cationic proteins were observed in the course of the study: the maximum rise of stimulation indices was observed on the 14th day after immunization and a less pronounced rise on the 28th day. The peak of agglutinating antibody production was on the 21st day after immunization, complement-binding antibodies – on the 21st-28th day, less intensive rise in titers of agglutinating and complement-binding antibodies was noted on the 69th day. In these terms, a decrease in the stimulated activity of neutrophil cationic proteins was observed. The peak activity of non-enzymatic cationic proteins of neutrophils was found to be ahead of the peak of humoral immune response.

Role of NaV1.7 in postganglionic sympathetic nerve function in human and guinea-pig arteries.

NaV1.7 plays a crucial role in inducing and conducting action potentials in pain-transducing sensory nociceptor fibres, suggesting that NaV1.7 blockers could be effective non-opioid analgesics. While SCN9A is expressed in both sensory and autonomic neurons, its functional role in the autonomic system remains less established. Our single neuron rt-PCR analysis revealed that 82% of sympathetic neurons isolated from guinea-pig stellate ganglia expressed NaV1.7 mRNA, with NaV1.3 being the only other tetrodotoxin-sensitive channel expressed in approximately 50% of neurons. We investigated the role of NaV1.7 in conducting action potentials in postganglionic sympathetic nerves and in the sympathetic adrenergic contractions of blood vessels using selective NaV1.7 inhibitors. Two highly selective NaV1.7 blockers, GNE8493 and PF05089771, significantly inhibited postganglionic compound action potentials by approximately 70% (P<0.01), with residual activity being blocked by the NaV1.3 inhibitor, ICA121431. Electrical field stimulation (EFS) induced rapid contractions in guinea-pig isolated aorta, pulmonary arteries, and human isolated pulmonary arteries via stimulation of intrinsic nerves, which were inhibited by prazosin or the NaV1 blocker tetrodotoxin. Our results demonstrated that blocking NaV1.7 with GNE8493, PF05089771, or ST2262 abolished or strongly inhibited sympathetic adrenergic responses in guinea-pigs and human vascular smooth muscle. These findings support the hypothesis that pharmacologically inhibiting NaV1.7 could potentially reduce sympathetic and parasympathetic function in specific vascular beds and airways. KEY POINTS: 82% of sympathetic neurons isolated from the stellate ganglion predominantly express NaV1.7 mRNA. NaV1.7 blockers inhibit action potential conduction in postganglionic sympathetic nerves. NaV1.7 blockade substantially inhibits sympathetic nerve-mediated adrenergic contractions in human and guinea-pig blood vessels. Pharmacologically blocking NaV1.7 profoundly affects sympathetic and parasympathetic responses in addition to sensory fibres, prompting exploration into the broader physiological consequences of NaV1.7 mutations on autonomic nerve activity.

Experimental transmission of a novel relapsing fever group Borrelia harbored by Ornithodoros octodontus (Ixodida: Argasidae) in Chile.

Tick-borne relapsing fever spirochetes of genus Borrelia thrive in enzootic cycles involving Ornithodoros spp. (Argasidae) mainly, and rodents. The isolation of these spirochetes usually involves a murine model in which ticks are fed and the spirochetes detected in blood several days later. Such an experiment also demonstrates that a given species of tick is competent in the transmission of the bacteria. Here, soft ticks Ornithodoros octodontus were collected in Northern Chile with the objective to experimentally determine its capacity to transmit a Borrelia sp. detected in a previous study. Two Guinea pigs (Cavia porcellus) were used to feed nymphs and adults of O. octodontus and the spirochetes in blood were inspected by dark-field microscopy and nested PCR. Although spirochetes were not seen in blood, DNA was detected in only one animal 11 days after the ticks were fed. Genetic sequences of Borrelia flaB, clpX, pepX, recG, rplB, and uvrA genes retrieved from DNA extraction of positive blood were employed to construct two phylogenetic analyses. On the one hand, the flaB tree showed the Borrelia sp. transmitted by O. octodontus clustering with Borrelia sp. Alcohuaz, which was previously detected in that same tick species. On the other hand, concatenated clpX-pepX-recG-rplB-uvrA demonstrated that the characterized spirochete branches together with "Candidatus Borrelia caatinga", a recently discovered species from Brazil. Based on the genetic profile presented in this study, the name "Candidatus Borrelia octodonta" is proposed for the species transmitted by O. octodontus. The fact that spirochetes were not observed in blood of guinea pigs, may reflect the occurrence of low spirochetemia, which could be explained because the susceptibility of infection varies depending on the rodent species that is used in experimental models. Although the vertebrate reservoir of "Ca. Borrelia octodonta" is still unknown, Octodon degus, a rodent species that is commonly parasitized by O. octodontus, should be a future target to elucidate this issue.

A Review of Slaughterhouse Blood and its Compounds, Processing and Application in the Formulation of Novel Non-Meat Products

Animal blood has become of growing interest, and its functional and nutritional properties are being exploited. In recent years, several research papers related to the application of blood in food products have been published. The purpose of this review is to describe animal blood, its chemical composition, sampling, processing, preservation, and its application in various non-meat products. Bovine, pig and guinea pig blood has been used in the formulation of different foods such as chocolate, cookies, sausages, drinks, gummies, extruded products and consumed directly as a nutritional supplement, the compounds of interest being heme iron from hemoglobin, blood plasma and serum, bioactive proteins and peptides. However, animal blood residues have a high microbial load that is controlled in slaughterhouses. Likewise, the use of this by-product has shown an increase in hemoglobin levels in pregnant mothers and children with anemia who consume it. These fortified foods were high in protein and iron. The use of blood in different food matrices is a potential alternative to improve its nutritional quality, in addition to helping to reduce the levels of malnutrition and anemia in people.

Virus-inhibitory activity of the antigen complex of opportunistic pathogenic bacteria against SARS-CoV-2

Introduction. The antigen complex of opportunistic pathogenic bacteria (ACOPB) has a protective effect against avian influenza viruses, herpes virus type 2, and other viruses that cause acute respiratory viral infections. In the context of the COVID-19 pandemic, an important task is to find out whether ACOPB has a protective effect against SARS-CoV-2.&#x0D; The purpose of the study was to evaluate in vitro the ACOPB virus-inhibitory activity against the Dubrovka laboratory strain of SARS-CoV-2.&#x0D; Materials and methods. The study was performed using Vero cell line CCL-81, human peripheral blood mononuclear cells (PBMCs), mouse monoclonal anti-idiotypic antibodies structurally mimicking biological effects of human interferons (IFNs), the Dubrovka laboratory strain of SARS-CoV-2. The infectivity of the virus was assessed by two methods: by virus titration using cell cultures and the limiting dilution method when the results are assessed by a cytopathic effect; the second method was a plaque assay. The in vitro virus inhibition test was performed using the cell culture susceptible to SARS-CoV-2; the mixture containing a specific dose of the virus and a two-fold dilution of ACOPB was transferred to the cell culture after the ACOPB medication had interacted with the virus at 4C for 2 hours. The ACOPB virus-inhibitory activity against SARS-CoV-2 was assessed by the functional activity of / and IFN receptors (RIFN) in human PBMCs induced in vitro by ACOPB and the ACOPB mixture with the specific dose of SARS-CoV-2. The RIFN expression level was measured by the indirect membrane immunofluorescence test.&#x0D; Results. Hemagglutination assay using chicken, mouse, guinea pig, and human red blood cells was performed for detection of the SARS-CoV-2 inhibitory protein. The lysate of Vero CCL-81 cells infected with SARS-CoV-2 Dubrovka demonstrated the highest hemagglutination activity with guinea pig red blood cells and low titers of hemagglutination in the virus-containing fluid. The virus inhibition test in the Vero CCL-81 cell culture demonstrated that ACOPB inhibited 10 doses of SARS-CoV-2 Dubrovka with the titer 1 : 32, providing 100% protection of the cell culture for 8 days (the monitoring period). ACOPB induced / and RIFN expression on membranes of human PBMCs in in vitro cultures and decreased RIFN / and expression after its interaction with SARS-CoV-2 Dubrovka.&#x0D; Conclusion. The experimental studies including the virus inhibition test in the cell culture susceptible to SARS-CoV-2 Dubrovka and the indirect membrane immunofluorescence assay using monoclonal anti-idiotypic antibodies mimicking IFN-like properties demonstrated that ACOPB had both an immunomodulatory and a virus-inhibitory effect.

An extract from the culture of a thermophilic &lt;i&gt;Staphylococcus aureus&lt;/i&gt; strain suppresses allergic inflammation in the airways &lt;i&gt;in vivo&lt;/i&gt; and degranulation of mast cells and basophils &lt;i&gt;in vitro&lt;/i&gt;

Aim. To study the anti-allergic effects of ruzam, an extract from the culture of a thermophilic Staphylococcus aureus strain, in an in vivo model of asthma and its influence on degranulation of mast cells and basophils in vitro.Materials and methods. Allergic asthma in guinea pigs was reproduced by two intraperitoneal injections of ovalbumin followed by a series of inhalations of this antigen for 1.5 months. Ruzam (6 μg / kg) or a reference drug (sodium cromoglycate, 3 mg / kg) was administered daily via a nebulizer during the last 6 days of immunization. One day after completion of inhalations with ovalbumin and compared drugs, changes in the airways were assessed using cytological, morphometric, and histologic methods. Rabbit blood basophils and rat peritoneal mast cells were used to determine the effect of ruzam on IgE-independent degranulation induced by the compound 48 / 80 in vitro. The effect of ruzam was compared with that of hydrocortisone hemisuccinate. Basophils from the blood of ovalbumin-sensitized guinea pigs were used to evaluate the effect of the drug on IgE-dependent degranulation induced by ovalbumin. Granules of mast cells and basophils were detected by alcian blue staining to calculate the degranulation index.Results. In the asthma model, ruzam reduced the degree of airway obstruction by increasing the bronchoalveolar lavage volume returned and suppressed neutrophilic and eosinophilic inflammation, while mobilizing other effector cells of the anti-pathogen immunity (lymphocytes and macrophages). Ruzam has proven to have a stronger anti-allergic effect than sodium cromoglycate by several parameters. At concentrations of 8.4–840 μg / ml, ruzam inhibited degranulation of mast cells and basophils, induced by the compound 48 / 80, equally to hydrocortisone hemisuccinate (10–3 M). At concentrations of 280 and 420 μg / ml, ruzam dose-dependently inhibited ovalbumin-induced degranulation of basophils in sensitized guinea pigs.Conclusion. The anti-allergic effect of ruzam was confirmed in test systems in vivo and in vitro. We speculate here that the TLR2 signaling pathway may be involved in biological and pharmacological effects of this drug.

A Novel Relapsing Fever Group Borrelia Isolated from Ornithodoros Ticks of the Brazilian Caatinga

Tick-borne relapsing fever group (RFG) borreliosis remains neglected as a human disease and little is known on its maintenance in ticks and vertebrates, especially in South America. Therefore, this study investigated borrelial infection in Ornithodoros ticks collected in rodent-inhabited rock formations in the Brazilian semiarid region, within the Caatinga biome. Collected ticks (Ornithodoros rietcorreai and Ornithodoros cf. tabajara) were allowed to feed under laboratory conditions on guinea pigs, which had blood samples examined daily by dark-field microscopy. No spirochetes were visualized in the blood of any of four O. rietcorreai-infested guinea pigs. Contrastingly, spirochetes were visualized between 9 and 39 days after tick feeding in the blood of three guinea pigs, each infested with O. cf. tabajara ticks from a different locality. Guinea pig infection was confirmed by passages into experimental animals and by generating DNA sequences of Borrelia spp. from the blood of spirochetemic guinea pigs. Three O. cf. tabajara populations were infected by the same borrelial organism, which was characterized as a novel RFG agent (named as 'Candidatus Borrelia caatinga') based on 10 Borrelia loci (rrs, flaB, glpQ, gyrB, clpX, pepX, pyrG, recG, rplB and uvrA). We demonstrated that O. cf. tabajara is a competent vector of the novel Borrelia sp. isolates, although none of the infected rodents developed clinical illness.

THE ROLE OF NITRIC OXIDE AND PEROXYNITRITE IN THE ERYTHEMAL PERIOD OF ULTRAVIOLET-INDUCED SKIN DAMAGE.

The aim: To study the activity of inducible NO synthase in the blood, the content of NO metabolites and peroxynitrite in the skin and blood of guinea pigs during erythema periods after local ultraviolet skin irradiation. Materials and methods: The studies were carried out on 24 guinea pigs, subjected to local ultraviolet irradiation. The control group consisted of intact guinea pigs (n=6). After 2, 4 hours, on the 3rd, 8th day, the activity of inducible NO synthase (in blood), the content of total NO metabolites, nitrite anion, nitrates, peroxynitrite (in blood and skin) were determined. Results: An increase in the level of peroxynitrite in the blood and of all NO metabolites in the skin was noted 2 hours after irradiation. After 4 hours activation of inducible NO synthase in the blood was observed, accompanied by an increase in the content of all NO metabolites and peroxynitrite both in the blood and in the skin. On the 3rd day high levels of excessive synthesis of NO and peroxynitrite in the blood and skin were also revealed due to the induced expression of this enzyme in the blood. On the 8th day the activity of inducible NO synthase remained high in the blood, which led to the accumulation of all NO metabolites and peroxynitrite in the blood and skin. Conclusions: Thus, local ultraviolet irradiation of the skin of guinea pigs leads to the activation of inducible NO synthase in the blood and an increase in NO synthesis and peroxynitrite in the blood and skin.

Characteristics of changes in individual cytokines in the blood of guinea pigs under the conditions of the development of experimental allergic alveolitis = Характеристика змін окремих цитокінів у крові морських свинок за умов розвитку експериментального алергічного альвеоліту

Characteristics of changes in individual cytokines in the blood of guinea pigs under the conditions of the development of experimental allergic alveolitis = Характеристика змін окремих цитокінів у крові морських свинок за умов розвитку експериментального алергічного альвеоліту

The effect of the combination of thiotriazoline and corvitin on the values of some parameters of the cellular and humoral immune system during the experimental reproduction of allergic alveolitis under immobilization stress

The purpose of the study was to determine the effectiveness of the combination of corvitin and thiotriazoline for the correction of levels of T- and B-lymphocytes, circulating immune complexes (CIC) of the blood of male guinea pigs (GP) if experimental allergic alveolitis (EAA) and immobilization stress (IS) are combined. The study was performed on 50 male GP weighing 180-210 g. EAA was reproduced by the method of O.O. Orekhov, Yu.A. Kyrylov. The production of IS was carried out by the method of P.D. Horizontov. The content of T- and B-lymphocytes in the blood was determined by the method of E.F. Chernushenko, L.S. Kogosova. CIC were detected by the method of Haskova V., Kaslik J., Math J., Matejckava M. The withdrawal from the experiment was carried out by the method of decapitation after previous administration of nalbuphine hydrochloride in the experimental group (EG) of intact GP; in the EG of EAA and IS during the study days corresponding to the stages of development of IS: 1st – anxiety, 14th – resistance, 24th – exhaustion; EG of EAA with IS on the 24th day after pharmacocorrection with the above-mentioned combination of drugs. There was a progressive decrease in the level of T-lymphocytes by 33,47% (p&lt;0,01), 36,23% (p&lt;0,01) and 39,62% (p&lt;0,01), an increase in B-lymphocytes by 27,82% (p&lt;0,01), 131,13% (p&lt;0,01) and 44,37% (p&lt;0,01), an increase in CIC by 41,15 % (p&lt;0,01), 46,62% (p&lt;0,01), and 50,78% (p&lt;0,01) on the 1st, 14th and 24th days respectively, compared to the indicators of the control group under the combination of EAA and IS. An increase in T-lymphocytes by 40% (p&lt;0,01), a decrease in B-lymphocytes by 24,31% (p&lt;0,01), and a decrease in the level of CIC by 26,08% (p&lt;0,01) were observed in the EG using corvitin and tiotriazoline relative to the corresponding indicators of the EG with a combination of EAA and IS without pharmacological interventions on 24th day of the experiment. The results indicate an immunocorrective effect of the combination of corvitin and thiotriazoline which is manifested by activation of the cellular and suppression of the humoral link of immunity of GP exposed to EAA and IS.

The state of individual components of humoral immunity of guinea pig blood in the dynamics of the formation of experimental periodontitis and immobilization stress and correction of their disorders by thiocetam

The research found a rapid increase in the level of immunoglobulin M, starting from the 3 rd day of the experiment in comparison with the control values, which clearly indicates its activation in particular, and stimulation of humoral immunity in general. In the later stages (on the 5 th and 15 th days) of the formation of experimental periodontitis (EP) and immobilization stress (IS), an increase of this indicator by 93.5% and 94.2% was found, respectively, against group I guinea pigs (p≤0, 05).Regarding the content of immunoglobulin G in the blood, the highest growth rates were recorded in the fourth group of guinea pigs with EP and IS (on the 15 th day), which indicates a direct dependence of the duration of damaging factors on the level of these indicators and the body's ability to the protective response. The results of the treatment showed the effect and the reduction of the activity of the studied classes of immunoglobulins. Thus, the use of thiocetam led to a significant decrease in the content of Ig M and G in the blood, respectively, by 33.8% (p1≤0.05) and 41.7% (p1≤0.05) in EP and IS compared with the group of animals were not exposed to this drug, which indicates the immunocorrective effect of this drug on the studied tests.

Preparation of bovine viral diarrhea disease virus 1 virus-like particles and evaluation of its immunogenicity in a guinea pig model

In order to obtain virus-like particles (VLPs) for prevention of bovine viral diarrhea virus 1 (BVDV-1), the C-Erns-E1-E2 region was cloned into a pFastBacDaul vector for generating the recombinant Bacmid-BVDV-1 in DH10Bac Escherichia coli. The recombinant baculovirus Baculo-BVDV-1 was produced by transfecting the Sf9 cells with Bacmid-BVDV-1. The expressed protein and the assembled VLPs were determined by immunofluorescence, Western blotting and electron microscopy. Guinea pigs were immunized with inactivated VLPs coupled with the Montanide ISA-201 adjuvant. The immunogenicity of VLPs was evaluated by monitoring the humoral immune response with neutralizing antibody titer determination, as well as by analyzing the cell-mediated immune response with lymphocyte proliferation assay. The protective efficacy of VLPs was evaluated by challenging with 106 TCID50 virulent BVDV-1 strain AV69. The results showed that the recombinant Baculo-BVDV-1 efficiently expressed BVDV structural protein and form VLPs in infected Sf9 cells. The immunization of guinea pigs with VLPs resulted in a high titer (1:144) of neutralizing antibody, indicating an activated cellular immunity. Significantly lower viral RNA in the blood of the post-challenged immunized guinea pigs was observed. The successful preparation of BVDV VLPs with insect cell expression system and the observation of the associated immunogenicity may facilitate further development of a VLPs-based vaccine against BVD.

Comparative characteristics of different methods of treatment of microsporia

Microsporia is the most common disease of fungal etiology, most often caused by the pathogen Microsporum canis. Treatment of this disease requires a comprehensive approach, because the disease is dangerous to humans. Therefore, in the treatment of microspores, it is important not only to carry out therapeutic measures, but also to prevent the spread of the pathogen of the fungus in the environment and increase the immune status of the organism in the fight against infection. The main source of infection is cats. In order to determine the effectiveness of treatment of microsporia by various methods, studied of the blood and skin of guinea pigs infected with the pathogen M. canis. Sick animals were divided into three groups. The first group was treated with the systemic antifungal itraconazole and topical treatment with a solution of clotrimazole. Treatment of the second group was performed with a topical antifungal agent (1% solution of clotrimazole) with vaccination with the antifungal vaccine “Vakderm”. For the third group, the developed drugs were used – antifungal agent “Micromar” and immunostimulant “Biogluk”. During treatment, hematological and immunological blood researches and histological skin examinations were performed. During treatment with antifungal drugs (itraconazole and clotrimazole) the number of leukocytes decreases from 11.13 ± 0.72 to 7.13 ± 0.22, rod-shaped neutrophils from 15.76 ± 1.29 to 5.50 ± 0.76, and segmental increases from 12.17 ± 1.47 to 24.17 ± 2.27, decreases ESR from 5.67 ± 0.67 to 2.33 ± 0.42, which occurs when inhibiting the inflammatory response of the organism to infection. Thrombocytopenia (from 231.17 ± 7.60 to 184.33 ± 7.65) and eosinophilia (from 2.70 ± 0.73 to 7.33 ± 1.33) are also noted. There is a slight increase in T-helpers and a decrease in T-suppressors. Histologically, the infiltration of the dermis by histiocytes and eosinophils under skin persists. In the treatment of microsporia by treatment with 1 % solution of clotrimazole and vaccination with the vaccine “Vakderm” the results of studies showed that the number of leukocytes decreases from 11.13 ± 0.72 to 5.35 ± 0.31 (P &lt; 0.01), rod-shaped neutrophils from 15.76 ± 1.29 to 7.67 ± 0.56, and segmental increases from 12.17 ± 1.47 to 22.17 ± 0.91 (P &lt; 0.001), decreases). The number of T-helpers is increasing. The histological picture on day 7 is characterized by hyperkeratosis, and on day 14 the hyperemia of the basal layer of the epidermis persists. When using the antifungal drug “Micromar” and immunostimulant “Biogluk” the results of studies showed that the number of leukocytes decreases from 11.13 ± 0.72 to 6.95 ± 0.10, rod-shaped neutrophils from 15.76 ± 1.29 to 6.17 ± 0.65, and segmental increases from 12.17 ± 1.47 to 22.00 ± 0.86, decreases ESR from 5.67 ± 0.67 to 2.17 ± 0.31. Increases the number of T-helpers and the number of natural killers and T-suppressors is gradually decreasing. Histological changes are presented in the form of dilation of blood vessels and visualization of single erythrocytes in the dermis on day 14 of treatment.

Evaluation of endogenous intoxication indicators in the dynamics of experimental periodontitis and immobilization stress

The aim of our work was to investigate the peculiarities of changes in endogenous intoxication rates: medium mass molecules (MMM) and erythrocyte intoxication index (EII) in the blood of guinea pigs in the formation of combined pathology - experimental periodontitis (EP) and immobilization stress (IS).The results of biochemical studies showed that at all stages of development of combined pathology - experimental periodontitis and immobilization stress there is a consistent increase in the degree of endogenous intoxication, which dominated on the 15th day of the experiment, namely increased content of medium mass molecules (МMМ254 , MMM280) and EII in the blood by 58.5% (p≤0.05), 80.3% (p≤0.05) and 95.7% (p≤0.05), respectively, compared with the first group of animals is a clear manifestation of the formation of endogenous intoxication. These changes are obviously the result of insufficient elimination of toxic products on the background of the inflammatory process in periodontal tissues and stress.

Dynamics of morphological, immunological and histological changes in microsporіа in guinea pigs

Microsporіа affect different species of animals and humans. The high contagiousness of the pathogen determines the relevance of research into this disease. Microsporum canis is the pathogen that most often causes microsporia. Weakened functions of the immune system and violation of the epithelial barrier of the skin are a favourable factor that causes microspores. The main source of infection is cats, which are involved in the storage and transmission of the pathogen. To clarify the dynamics of morphological, immunological and histological changes in microsporia, blood and skin studies of guinea pigs infected with M. canis were carried out. The animals were divided into two groups of 6 guinea pigs (healthy and sick). Test material (blood and skin) was taken from clinically healthy and sick animals 21 and 42 days after infection. The number of erythrocytes and leukocytes was determined by counting them in the Goryaev chamber, the hemoglobin content – by the method of cyanide hemoglobin. The leukogram was derived based on the counting and differentiation of 200 leukocyte cells in blood smears. Material for histological examination (pieces of skin) was fixed in 10–12% cooled solution of neutral formalin, followed by pouring in paraffin according to the scheme proposed by G. A. Merkulov. The obtained results demonstrated that leukocytosis developed in guinea pigs with microsporia on the 21st and 42nd days; the number of rod-shaped neutrophils increased, that of segmental neutrophils decreased, and that of ESR increased. The immune response to the course of microsporia was manifested in an increase in the percentage of T-lymphocytes, T-suppressors and a decrease in T-helper cells and an increase in T-killers compared with healthy animals. Histological examination showed that on the 21st day after infection, hyphae and spores of the fungus M. canis were localized in the skin. There is swelling of the dermis, stratification of collagen fibers and the accumulation of inflammatory infiltrates around the hair follicles. On the 42nd day, the infiltration spread and dystrophic changes in the skin occurred in the form of desquamation of the epidermis and the formation of acanthosis and hyperkeratosis on the surface of the dermis. The conducted research will allow further assessment of the course of microsporia under the action of various drugs and help establish the most effective method of treatment.

DNA methylome signatures of prenatal exposure to synthetic glucocorticoids in hippocampus and peripheral whole blood of female guinea pigs in early life

Synthetic glucocorticoids (sGC) are administered to women at risk of preterm delivery, approximately 10% of all pregnancies. In animal models, offspring exposed to elevated glucocorticoids, either by administration of sGC or endogenous glucocorticoids as a result of maternal stress, show increased risk of developing behavioral, endocrine, and metabolic dysregulation. DNA methylation may play a critical role in long-lasting programming of gene regulation underlying these phenotypes. However, peripheral tissues such as blood are often the only accessible source of DNA for epigenetic analyses in humans. Here, we examined the hypothesis that prenatal sGC administration alters DNA methylation signatures in guinea pig offspring hippocampus and whole blood. We compared these signatures across the two tissue types to assess epigenetic biomarkers of common molecular pathways affected by sGC exposure. Guinea pigs were treated with sGC or saline in late gestation. Genome-wide modifications of DNA methylation were analyzed at single nucleotide resolution using reduced representation bisulfite sequencing in juvenile female offspring. Results indicate that there are tissue-specific as well as common methylation signatures of prenatal sGC exposure. Over 90% of the common methylation signatures associated with sGC exposure showed the same directionality of change in methylation. Among differentially methylated genes, 134 were modified in both hippocampus and blood, of which 61 showed methylation changes at identical CpG sites. Gene pathway analyses indicated that prenatal sGC exposure alters the methylation status of gene clusters involved in brain development. These data indicate concordance across tissues of epigenetic programming in response to alterations in glucocorticoid signaling.

Pharmacokinetic study of midazolam and α-hydroxymidazolam in guinea pig blood and hair roots after a single dose of midazolam

The appearance of midazolam (M) and its metabolites into the hair root following a single administration was examined by following the time course of M and α-hydroxymidazolam (αHM) in hair roots and blood from guinea pigs. The back hair of guinea pigs was shaved before drug administration and before each sampling, and hair roots (3–5 mm) were plucked at 5, 15, and 30 min, 1, 2, 4, 6, 10, 24, 48, 72, 96, 120, 144 h, and 7, 14, 21, and 28 days. The kinetic parameters of M and αHM in guinea pig blood and hair roots were determined for three doses (5, 10, and 25 mg/kg). Comparisons of drug time course between hair roots and blood indicated an association between drug concentrations in the hair root and the blood. M and αHM entered the hair root within 5 min after a single exposure. The detection windows were also longer for the hair root than for the blood. Consequently, the hair root can be a valuable specimen in acute poisonings or drug-facilitated crime (DFC) cases, if other matrices are unavailable, or if blood and urine results are negative. Hair shafts (with hair roots) were plucked at 28 days and segmented. The concentrations of M and αHM were lower in the hair shafts than in the hair roots. The concentrations of the metabolite αHM in hair shafts were barely detectable. The concentrations of M and αHM in the hair root showed a moderate correlation with dose. Comparison of drug levels in hair roots between the washed group and the unwashed group indicated a generally stable percentage between the washed and unwashed groups of 40–60 % during the entire time course. This indicates that drugs are likely to be immobilized in the hair root at the beginning of the incorporation process.

Practical aspects on identification, cultivation and characteristics of varicella-zoster virus isolates

Until now, it has been considered that infectivity of the varicella-zoster virus (VZV) is closely related to target cell, and newly formed virus is not released into the culture medium. It is also known that it is hard to grow VZV in cell cultures, due to its slow replication rate and a limited range of sensitive cell cultures. In addition, VZV isolation depends on type of cell culture used, nature of clinical material, presence of viable virus and transport time. Objectives. To study production of infectious extracellular VZV in various cell cultures. Materials and methods. Eight cell cultures were used, including human embryonic diploid lung cells and human embryonic dermomuscular tissue (KM-27), as well as continuous human and monkey cell lines. Crusts detached from vesicular lesions were used as clinical isolates, which were placed into cryo-vials added with transport medium and transferred in liquid nitrogen. VZV infectivity was assessed in cell cultures by using hemo-adsorption assay with erythrocyte suspension isolated from guinea pig or human zero group blood and confirmed by indirect immunofluorescence with polyclonal sera from varicella or herpes zoster convalescents. Results. There were examined 27 clinical samples consisting of crusts from vesicular lesions isolated from patients with chickenpox, as well as one sample from 63-year old patient with exacerbated recurrent herpes zoster. Primary infection with clinical isolates was performed on diploid human lung embryo cells (HLEC) at low temperature. It was found that clinical samples collected within day 1–18 inclusive after the onset of skin eruption were able to induce cytopathic effects in HLEC cell monolayer such as cytolysis around dermal crusts. Specificity of cytopathic effect was confirmed by using real-time polymerase chain reaction (RT-PCR). Viral antigens were prepared on 7 cell lines infected with the laboratory strain Ellen VZV (USA) to assess the immune sera. A high anti-VZV specificity of mouse sera was detected by ELISA while all the lysates of infected cell lines were used as the solid-phase sorbent. In experiments on VZV reproduction demonstrated that extracellular virus was released into the culture medium starting from day 1 after infection of target cells, and infectivity of the virus-containing fluid ascends during further cultivation.