Modern Methods of DNA Sequencing (Scientific Review)

Rapidly developing sequencing technologies have provided the possibility for identification of a DNA nucleotide sequence of a whole individual human genome just in a couple of weeks. Working capacity of some sequencers is already measured in thousands of milliards of base pairs per an operating cycle. Reviewed have been the basic principles and analytical potential of the modern methods for DNA sequencing which are nominally subdivided into three major types: classical - capillary electrophoresis sequencing and pyrosequencing, novel (Next Generation Sequencing - NGS) - simultaneous sequencing of millions of DNA fragments, each one of which is a cluster containing many thousands or hundreds of thousands of their clones - high-performance pyrosequencing, cyclic ligase and semiconducting sequencing, molecular-cluster-based sequencing using fluorescent-labeled precursors; and cutting-edge methods - (Next-Next Generation Sequencing - NNGS) - the ones that read millions of single DNA fragments without pre-cloning

THE INFLUENCE OF COLLOID SILVER NANO-PARTICLES UPON IMMUNOBIOLOGIC PROPERTIES OF PLAGUE MICROBE CAPSULAR ANTIGEN

This study shows that Yersinia pestis capsular antigen F1 conjugated to the CS nano-particles demonstrates apparent immunogenic effects. When injected to the test animals it was able to stimulate the production of specific antibodies under smaller antigenic load, it caused activation of the genetic apparatus in the immune system cells, and it stimulated hyperplastic processes in the lymphoid organs. Solutions of CS plus antigens appeared to be stable and caused neither general nor local responses in the test animals; they were easy to make use of, thus demonstrating their potentialities to be exploited as adjuvants in immunologic practices

Extensive Studies of Legionella Strains Isolated from the Environmental Objects during Preparation and Holding of Mass Events in the Russian Federation in 2013-2014

Objective of the work was to conduct an extended study of Legionella strains isolated from epidemiologically significant environmental objects during the preparation and conduct of mass events in the territory of the Russian Federation in 2013-2014 . Materials and methods. Studied were 53 strains of Legionella pneumophila, isolated from epidemiologically significant objects during the preparation and conduct of a number of mass events (ME): XXVII World Summer Universiade, XXII Olympic Winter Games and XI Paralympic Winter Games, Sochi; Summer Health Promotion Campaign in 2014, Republic of Crimea; IV Caspian Summit, Astrakhan, 2014. Strains were analyzed using multilocus sequencing, time-of-flight mass spectrometry (MALDI-ToF-MS) and atomic force microscopy. Legionella strains were typed by multilocus sequencing in accordance with the algorithm of the European Legionellosis Research Group “Sequence-Based Typing protocol for epidemiological typing of L. pneumophila". Results and discussion. Strains of L. pneumophila, legionellosis agent, were isolated in the territory of the Republic of Crimea, Moscow, Kazan, Sochi, Astrakhan and characterized. According to the results of slide agglutination, 17 L. pneumophila strains were assigned to 1 serogroup , 37 - to 2-14 serogroups. Based on the data obtained by multilocus sequencing, in accordance with the algorithm of the European Working Group on Legionellosis Surveillance, allelic profiles of all the studied L. pneumophila strains were identified; their belonging to 7 sequence types was established. Using the method of time-of-flight mass spectrometry, legionella strains were characterized, their protein profiles were studied, and a database was formed. Using the method of scanning probe microscopy, information was obtained on the morphology of the cells of 18 legionella strains and the features of their surface structures. Using the methods of multilocus sequencing, time-of-flight mass spectrometry, and atomic force microscopy, molecular-genetic, proteomic, and morphometric features of Legionellosis pathogen strains that circulate in epidemiologically significant sites in the Russian Federation were determined

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<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<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<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

Olfactory transport efficiency of the amorphous and crystalline manganese oxide nanoparticles

The ability to deliver particulated xenobiotics and therapeutic drugs directly from the nasal cavity to the central nervous system, bypassing the hemato-encephalic barrier, determines a high importance of investigation of factors influencing this process. It was shown that the bioavailability of solid particles is influenced by their size and surface charge. At the same time, the impact of a crystal structure (crystalline/amorphous) has been poorly investigated. In this study, using sexually mature male C57BL/6J mice, we analyzed the efficiency of the nose-to-brain transport of crystalline and amorphous manganese oxide nanoparticles. T1-weighted magnetic resonance imaging (MRI) was used to evaluate the accumulation of manganese nanoparticles in olfactory bulb (OB) and olfactory epithelium (OE). So, it has been established that amorphous particles have higher accumulation rate in OE and OB in comparison with crystalline particles after their intranasal administration. The unequal ability of amorphous and crystalline particles to overcome the mucosal layer covering the OE may be one of the possible reasons for the different nose-to-brain transport efficiency of particulated matter. Indeed, the introduction of mucolytic (dithiothreitol) 20 minutes prior to intranasal particle application did not influence the accumulation of amorphous particles in OE and OB, but enhanced the efficiency of crystalline nanoparticle entry. Data on the different intake of amorphous and crystalline nanoparticles from the nasal cavity to the brain, as well as the evidence for the key role of the mucosal layer in differentiating the penetrating power of these particles will be useful in developing approaches to assessing air pollution and optimizing the methods of inhalation therapy

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<0.01); pain by 35% at 1 month, by 32% at 3 months, and by 36% at 6 months (p<0.01); stiffness, by 37, 38, and 39% (p<0.01); and functional failure by 29, 25, and 23%, respectively (p<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<0,01); боли – через 1 мес на 35%, через 3 мес на 32% и через 6 мес на 36% (р<0,01); скованности – на 37, 38 и 39% (р<0,01); функциональной недостаточности – на 29, 25 и 23% соответственно (р<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

Эффективность хондроитина сульфата у пациентов с остеоартритом коленных и тазобедренных суставов

Currently chondroitin sulfate (CS) is the most studied drug from the group of «chondroprotectors». The article shows the role of CS in the treatment of osteoarthritis (OA): mechanism of action, clinical efficacy and safety in patients with OA with comorbidity. Evidence for the effectiveness and safety of the use of CS has been analyzed. The place of this drug in Russian and international clinical guidelines for the management of patients with OA is discussed.In domestic practice, the parenteral form of CS is often used in patients with OA. The parenteral form of CS for intramuscular and intra-articular administration (Chondroguard®) has a number of advantages, including the rapid onset of the effect. This enables reducing the dose in a short time or completely abandon the use of non-steroidal anti-inflammatory drugs (NSAIDs), which is extremely important for patients with OA with comorbidity. Chondroguard® can be recommended for initial use in exacerbations of chronic joint and back pain in OA, including in patients with relative and absolute contraindications for NSAID therapy.Хондроитина сульфат (ХС) – наиболее изученный на сегодняшний день препарат из группы «хондропротекторов». Описаны механизм действия, клиническая эффективность и безопасность ХС у больных остеоартритом (ОА) с коморбидностью. Проанализирована доказательная база эффективности и безопасности применения ХС. Обсуждается место препарата в отечественных и международных клинических рекомендациях по ведению пациентов с ОА.В отечественной практике у больных ОА часто используется парентеральная форма ХС. Парентеральная форма ХС для внутримышечного и внутрисуставного введения (Хондрогард®) имеет ряд преимуществ, среди которых быстрое наступление эффекта. Это позволяет в короткие сроки снизить дозу или полностью отказаться от приема нестероидных противовоспалительных препаратов (НПВП), что крайне важно для больных ОА с коморбидностью. Хондрогард® может быть рекомендован для стартового применения при обострениях хронической боли в суставах и спине при ОА, в том числе у пациентов с относительными и абсолютными противопоказаниями для назначения НПВП

Азотсодержащие вещества в снеге районов падения ступеней ракеты-носителя «Протон» в 2009–2019 гг.

The article presents analysis of the snow pollution caused by flights of the Proton launch vehicles launched from the Baikonur cosmodrome in 2009–2019. Data on 1477 snow samples collected in areas in Central Kazakhstan, south­east of Western Siberia, and north­east of the Altai are summarized to assess the pollution. In the uninhabited areas in Central Kazakhstan, where rocket fuel is spilled, pollution of snow was detected at 18 fall sites at a distance of up to 10 m from the fragments of the stages. The background value of pH is estimated as 6.6±0.9. The following nitrogen-­containing compounds (mg/l) were found in snow taken in the unpolluted territories of Central Kazakhstan: NO3­ (2.3±3.4), NH4+ (0.75±0.98) and to a lesser extent NO2– (0.015±0.019); asymmetric dimethylhydrazine and nitrosodimethylamine were not detected. The chemical composition of snow in the fall areas of the second stage of the Proton launch vehicle is in a good agreement with the background level of nitrogen­-containing substances of natural origin. There were no traces of the rocket fuel. Thus, in the snow on the Ketsko­Tym plain (south­east of Western Siberia) and north­east of Altai, the background content of nitrogen-­containing substances is equal, respectively (mg/l): NO3– (0.36±0.28 and 0.47±0.59); NH4+ (< 0.05 and 0.20±0.27); NO2– (0.048±0.016 and 0.027±0.073), which is determined by regional features and distance from sources of nitrogen­-containing substances. The data obtained allow us to conclude that the areas of the rocket falls on the above territories are not an environmental hazard to the environment by the concentration of nitrogen-­containing substances in the snow.Обобщены данные по 1477 пробам снега из районов падения ступеней ракеты-носителя «Протон» в Центральном Казахстане, на юго-востоке Западной Сибири и северо-востоке Алтая. В безлюдных территориях Центрального Казахстана, куда попадает ракетное топливо, достоверное загрязнение снега установлено на расстоянии не более 10 м от фрагментов падения первой ступени. Химический состав снега районов падения второй ступени (юго-восток Западной Сибири и северо-восток Алтая) отражает фоновые уровни азотсодержащих соединений природного происхождения и показывает отсутствие компонентов ракетного топлива. Очевидно, что районы падения и первой, и второй ступеней ракет-носителей «Протон», запускаемых с космодрома Байконур, не представляют собой зоны экологического бедствия

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