Development of the PCR Assay with Universal Primers for the Detection of Different <i>tcpA</i> Gene Variants

Developed is the PCR assay for the detection of the structural genes of toxin co-regulated adhesion piluses - tcpA of different types. Determined are the universal primers, the usage of which provides for the detection of the stated above genes in V. cholerae of various serogroups. With the help of this PCR assay identified is a new variant of tcpA gene in toxigenic cholera vibrio of non-O1/non-O139 serogroup

Taxonomic Position of Entomoparasitic Nematodes Isolated from Fleas in Gorno-Altai High-Mountain Natural Plague Focus

Objective of the study was to investigate entomoparasitic nematodes from rodent fleas, collected in Gorno-Altai high-mountain natural plague focus in 2016–2017, with identification of their taxonomic status. Materials and methods. Given are the results of morphological and genetic analysis of entomoparasitic nematodes – parasites of fleas Amphipsylla primaris primaris living on flat-headed high-mountain voles Alticola strelzovi, fleas Rhadinopsylla li transbaicalica of Daurian pika Ochotona dauurica, fleas Rhadinopsylla dahurica, collected at the burrows of the rodents. Results and conclusions. Identified have been nucleotide sequences of ITS2 spacer region of ribosomal operon in entomoparasitic nematodes of three flea species: Amphipsylla primaris primaris, Rhadinopsylla li transbaicalica, and Rhadinopsylla dahurica. Based on comparative phylogenetic analysis and identified high percentage of homology of the stated nucleotide sequences (92–99 %), appurtenance to Rubzovinema spp. species and close relation to earlier proposed by us multi-host species Rubzovinema polyxenica from C. tesquorum, A. rossica, and C. secundus fleas from Volga-Ural steppe plague focus has been established. For the first time ever separate branch of evolution of entomoparasitic nematodes – flea parasites, represented by Rubzovinema ssp., has been identified. Determined has been wide spread of Rubzovinema ssp. nematodes in biocenoses of steppe zone of Russia across Precaspian territory and Altai Mountains

Effect of <i>Yersinia pestis</i> on the Soil Nematodes <i>Panagrolaimus</i> sp. from the Gorno-Altai High-Mountain Focus of Plague

The aim of the work was to study interaction of Yersinia pestis with soil nematodes isolated on the territory of the Gorno-Altai high-mountain plague focus. Materials and methods. We used the fluorescent Y. pestis strain KM2083, a derivative of the natural strain of the 4.ANT phylogenetic line, the antique biovar of the main subspecies, and a nematode culture isolated in the same area of the Gorno-Altai plague focus. The taxonomy of nematodes was determined by the region of the 18S rRNA gene; phylogenetic analysis was performed using the Maximum Likelihood method based on the Tamura-Nei model in the Mega 7.0 software. The interaction of the Y. pestis KM2083 strain and the nematodes was studied during cultivation on a solid NGM agar medium. Nematodes were observed using microscopes Stemi-2000C (Carl Zeiss, Germany) and Axio Imager Z2 (Carl Zeiss, Germany). Results and discussion. It has been established that the nematodes from the Gorno-Altai high-mountain plague focus used in the work belong to the genus Panagrolaimus. Cultivation of nematodes on the lawn of the Y. pestis strain of the main subspecies of antique biovar, the 4.ANT phylogenetic line for 24 hours did not lead to a reduction in the lifespan of nematodes compared to the control sample, which indicates the absence of toxicity of the used strain towards Panagrolaimus nematodes. On the cuticle of nematodes, the formation of a biofilm in the genital area and tail has been noted, and accumulations of fluorescent cells of the plague pathogen observed in the digestive tract. The data obtained can indicate the ability of nematodes to carry the plague pathogen in the soil biocoenosis

Efficacy and safety of diacerein in patients with knee osteoarthritis

Diacerein (D) belongs to a class of symptomatic slow-acting agents, has an original mechanism of action, and is widely used as a diseasemodifying antirheumatic drug to treat osteoarthritis (OA) in Russia and many countries of the world. The ability of the drug to affect the main symptoms and progression of OA has been shown in a number of well-organized clinical trials.Objective: to evaluate the efficacy and safety of D in patients with knee OA.Patients and methods. An open-label trial evaluating the efficacy and safety of D (diaflex) in patients with knee OA was conducted in accordance with the multicenter program «Osteoarthrosis: Assessment of Progression in Real Clinical Practice». The trial included 80 patients of both sexes with Stage II–III knee OA; mean age, 60.8±6.8 years (47–75 years); mean body mass index, 31.8±5.9 kg/m2; disease duration, 10.3±5.7 years (2–30 years). The duration of the trial was 9 months (6 months of therapy and 3 months of follow-up).Results. There was a statistically significant reduction in visual analog scale pain on walking just 1 month after therapy initiation (57.1±9.7 and 44.7±13.9 mm; p&lt;0.0001) and a further significant improvement throughout the 6-month therapy. Pain did not increase after the drug was discontinued (the follow-up period was 3 months). The same pattern was observed in the assessment of the WOMAC index (pain during early therapy, 243.8±73.9; pain at the end of therapy, 137.5±78.9; stiffness, 97.8±41.1 and 57.7±38.6; functional failure, 875.8±250.4 and 525±305.7 respectively; p&lt;0.0001). Statistically significantly improved quality of life indicators measured by EQ-5D were noted throughout the follow-up period: 0.43±0.23 at the beginning of therapy, 0.61±0.14 at its end, and 0.63±0.11 at 3 months following treatment completion (p&lt;0.0001). By the time of therapy completion, 71.3% of the patients completely refused to take nonsteroidal anti-inflammatory drugs (NSAIDs). Both the patient and the physician evaluated the efficiency of treatment identically. By the end of therapy, 87.5% of the patients were observed to have improvement. Adverse reactions (ARs) were recorded in 10 (12.5%) patients and mainly associated with more frequent stools; ARs were not a cause of treatment interruptions or protocol deviations.Conclusion. Diaflex has a good symptomatic and anti-inflammatory effect: the therapy statistically significantly reduces pain, stiffness, and the need for NSAIDs and improves quality of life and joint function. The drug has a good safety profile and after-effects, which is seen at least 3 months after therapy discontinuation

Long-Term Persistence of <i>Yersinia pestis</i> in Association with Acanthamoeba castellanii in Experiment

The aim of the study was to test the feasibility of long-term survival and preservation of the properties of Yersinia pestis in association with soil amoeba Acanthamoeba castellanii. Materials and methods. Y. pestis strains and acanthamoeba isolated in the common area of the Gorno-Altai high-mountain plague focus were used for the study. The systematic affiliation of protozoa was determined through analyzing the 18S rRNA gene fragment sequencing data, followed by alignment with amoeba sequences from the NCBI GenBank database. A fluorescent Y. pestis strain was obtained by electroporation using the pTurboGFP-B plasmid. Co-cultivation was carried out in saline buffer in the absence of nutrients for the cells of plague pathogen. The influence of co-culturing with protozoa on Y. pestis properties was determined using microbiological, biological, and molecular-genetic methods. Results and discussion. The cell viability preservation for 22 months of the experiment in Y. pestis strain belonging to the main subspecies of the antique biovar, the 4.ANT phylogenetic line in co-culture with amoeba cells in the absence of additional nutrients has been established. Co-cultivation with amoebae did not lead to a change in the cultural, morphological, genetic and virulent properties of the plague pathogen strain. The data obtained confirm the possibility of using Acanthamoeba castellanii by the plague microbe to persist in soil biocenoses and open up the prospect of studying the mechanisms of plague pathogen surviving during extended inter-epizootic periods

OPTIMIZATION OF PRESENTATION AND CONSUMER CONTAINER OF ANTI-RABIES IMMUNOGLOBULIN OBTAINED FROM HORSE SERUM

We presented the data concerning optimization of presentation and consumer container of anti-rabies immunoglobulin obtained, from horse serum.. During the experiments, ampoules and. flasks were used for primary packaging. They were filled within 5 ml of the preparation of freeze-drying. Comparative analysis of physical, chemical and. biological properties and. molecular parameters of freeze-dried and initial forms of the immunoglobulin. was carried out. Freeze-drying was demonstrated to promote stabilization of its properties and. prevent emergence of fragments and. aggregates during anti-rabies immunoglobulin storage

Molecular-Genetic and Phenotypic Peculiarities of Plague Agent Strains Isolated in Vietnam

Objective of the study is to investigate phenotypic and molecular-genetic features and perform whole genome sequencing of Y. pestis strains isolated in Vietnam. Materials and methods. Studied were phenotypic and genotypic peculiarities of 20 plague agent strains isolated in different prefectures of Vietnam. Carried out was SNP-analysis of the strains, sequenced were genomes of 8 Y. pestis strains. Results and conclusions. Based on the results of studies of differential biochemical characteristics all the investigated strains were attributed to oriental biovar of the main subspecies of plague agent, which was confirmed by the presence of marker indel mutation – deletion of 93 bps in glpD gene. Investigated was also the plasmid composition of the strains. On the basis of the conducted genome sequencing and SNP-analysis appurtenance of 19 out of 20 strains under examination was determined. They belong to 1.ORI2v phylogenetic branch, relative to the strains isolated in Yunnan Province, China, which points to their common origin. Identified was a marker SNP and developed the method of SNP-typing for 1.ORI2v strains from Vietnam

Specific Appurtenance, Numbers, and Dynamics of Interaction of Acanthamoeba from Soils of Gorno-Altai High-Mountain Plague Focus with Yersinia pestis Strains

Objective of the study is to analyze the appurtenance, numbers, and dynamics of interaction of acanthamoeba from soils of GornoAltai plague focus with Yersinia pestis 367 strain, isolated in 2016 in enzootic territory of this focus. Materials and methods. Utilized were soil amoeba from Gorno-Altai high-mountain focus and the strain Y. pestis 367 of the main subspecies of antique biovar, isolated there in 2016. Determination of systematic relation of the isolated amoeba was carried out using PCR with genus specific primers and sequencing of the obtained PCR fragments followed by identification of nucleotide sequences against GenBank database. Localization of Y. pestis cells in acanthamoeba was performed using fluorescent antibody technique by means of Axio Imager Z2 (Carl Zeiss, Germany). Results and conclusions. For the first time ever established has been the presence of Acanthamoeba castellanii in soils of burrows of Marmota altaica in the numbers of up to 300000 cells/gr in Gorno-Altai high-mountain focus. Investigated has been the dynamics of interaction of these microorganisms. Preservation of the agent in vacuoles of endoplasmatic reticulum within 14 days has been revealed. It is an indicative of the possibility of Y. pestis persistence in amoeba of Acanthamoeba subspecies in soil biocoenosis of Gorno-Altai high-mountain plague focus

Оценка эффективности, переносимост и и безопасности внутрисуставного введения гиалуроновой кислоты у больных с о стеоартритом коленных суставов

Objective: to evaluate the efficacy, tolerability, and safety of intra-articular hyaluronic acid (hyalurom) in patients with knee osteoarthritis (OA).Patients and methods. A 6-month prospective trial enrolled 20 women aged 45–75 years (61±7 years) with primary knee OA of Kellgren–Lawrence grades II (85%) and III (15%) who needed nonsteroidal anti-inflammatory drugs (NSAIDs). The disease duration averaged 6.6±2.4 years. The mean body mass index was 33±5 kg/m2. Intra-articular administration of hyalurom was made; its cycle included 3 injections at a 1-week interval; a further follow-up was performed during 6 months. All the participants completed the trial.Results. In the first month of therapy, its effect was developed in the majority (75%) of the patients. There was a substantial reduction in total WOMAC scores by an average of 29% at 1 month, by 27% at 3 and 6 months (p&lt;0.01); pain by 35% at 1 month, by 32% at 3 months, and by 36% at 6 months (p&lt;0.01); stiffness, by 37, 38, and 39% (p&lt;0.01); and functional failure by 29, 25, and 23%, respectively (p&lt;0.01). The effect of therapy in most (80%) patients persisted throughout the follow-up period; only 10% of patients continued to take NSAIDs with the same frequency at 6 months; 60% used them on-demand, and 30% did not need NSAID therapy. No adverse reactions associated with the therapy performed were detected during the follow-up period.Conclusion. Hyalurom has a significant symptomatic effect and a good tolerability in the treatment of knee OA.Цель исследования – оценить эффективность, переносимость и безопасность внутрисуставного введения гиалуроновой кислоты (гиалуром) у больных с остеоартритом (ОА) коленных суставов.Пациенты и методы. В 6-месячном проспективном исследовании приняли участие 20 женщин 45–75 лет (61±7 лет) с первичным ОА коленных суставов II (85%) и III (15%) рентгенологических стадий по Kellgren–Lawrence, нуждающихся в приеме нестероидных противовоспалительных препаратов (НПВП). Длительность заболевания достигала в среднем 6,6±2,4 года. Средний индекс массы тела – 33±5 кг/м2. Проводилось внутрисуставное введение гиалурома, курс составлял 3 инъекции с интервалом в 1 нед, последующее наблюдение осуществлялось на протяжении 6 мес. Все участники завершили исследование.Результаты. У большинства пациентов (75%) эффект развивался в течение первого месяца. Выявлено значимое снижение суммарного индекса WOMAC – через 1 мес в среднем на 29%, через 3 и 6 мес на 27% (р&lt;0,01); боли – через 1 мес на 35%, через 3 мес на 32% и через 6 мес на 36% (р&lt;0,01); скованности – на 37, 38 и 39% (р&lt;0,01); функциональной недостаточности – на 29, 25 и 23% соответственно (р&lt;0,01). Эффект терапии у большинства пациентов (80%) сохранялся на протяжении всего периода наблюдения, к 6-му месяцу только 10% больных продолжали принимать НПВП с прежней частотой, 60% использовали их по требованию, 30% не нуждались в терапии НПВП. За период наблюдения не выявлено нежелательных реакций, связанных с проводимой терапией.Выводы. Гиалуром обладает значимым симптоматическим эффектом и хорошей переносимостью при ОА коленных суставов

ISOLATION OF GLICOPROTEID FROM THE FIXED RABIES VIRUS, STRAIN «MOSCOW 3253», AND CONSTRUCTING OF DOT-IMMUNOASSAY DIAGNOSTICUM ON ITS BASIS

Described here are the results of glicoproteid isolation from the fixed rabies virus, strain «Moscow 3253», using non-ionic detergent with subsequent chromatographic purification. The obtained antigen was demonstrated to be applicable as immunoreagent for construction of diagnosticum, by means of conjugation with colloid gold nanoparticles. The diagnosticum is meant for detection of specific antibodies in immune sera of horsesproducers, and in the preparation of anti-rabies immunoglobulin, in dot-immunoassay