The 35S U5 snRNP is generated from the activated spliceosome during In vitro splicing

Primary gene transcripts of eukaryotes contain introns, which are removed during processing by splicing machinery. Biochemical studies In vitro have identified a specific pathway in which introns are recognised and spliced out. This occurs by progressive formation of spliceosomal complexes designated as E, A, B, and C. The composition and structure of these spliceosomal conformations have been characterised in many detail. In contrast, transitions between the complexes and the intermediates of these reactions are currently less clear. We have previously isolated a novel 35S U5 snRNP from HeLa nuclear extracts. The protein composition of this particle differed from the canonical 20S U5 snRNPs but was remarkably similar to the activated B* spliceosomes. Based on this observation we have proposed a hypothesis that 35S U5 snRNPs represent a dissociation product of the spliceosome after both transesterification reactions are completed. Here we provide experimental evidence that 35S U5 snRNPs are generated from the activated B* spliceosomes during In vitro splicing

Studies of Sensitivity to Avian Flu Virus A/H5N1 in Chickens

) appear to be highly virulent for chickens. The chance of AFV infection of chickens in case of intranasal challenge is 20 times as great as in the case of peroral one, and 300 times as great as in the case of intragastral one, which bears evidence to higher sensitivity to AFV of the tissues of avian respiratory organs, in comparison with the tissues of gastro-intestinal tract. Therewith, primary target organ for virus in intranasal infected birds is their respiratory channel (mucous membrane of the nasal cavity in particular). Registered is the possibility of existence of fecal-nasal AFV transfer mechanism in chickens

Dissemination of Influenza A/H5N1 Virus after Intranasal Inoculation of Chickens

Studied was dissemination of avian influenza virus (AIV) in the organism of chickens after intranasal challenge with 10-100 LD50. The primary organ of accumulation of AIV A/H5N1 (A/Chicken/Kurgan/05/2005strain) is the respiratory tract (nasal mucosa), where the virus is registered in 18 hours after challenge. The accumulation of pathogen is observed in many organs and serum of chicken in 30-32 hours after challenge. The animals die in 54 hours, the concentration of virus reaches critical value in all studied samples. The highest AIV loads (7 lg of chicken embryo infective dose - EID50/g or ml) are registered in lungs, blood serum and kidneys of chicken. The results of AIV loads measuring using titration and real time RT-PCR show high degree of correlation (r=0.89)

Quantitative Proteome Profiling of C. burnetii under Tetracycline Stress Conditions

The recommended antibiotic regimen against Coxiella burnetii, the etiological agent of Q fever, is based on a semi-synthetic, second-generation tetracycline, doxycycline. Here, we report on the comparison of the proteomes of a C. burnetii reference strain either cultured under control conditions or under tetracycline stress conditions. Using the MS-driven combined fractional diagonal chromatography proteomics technique, out of the 531 proteins identified, 5 and 19 proteins were found significantly up- and down-regulated respectively, under tetracycline stress. Although the predicted cellular functions of these regulated proteins did not point to known tetracycline resistance mechanisms, our data clearly reveal the plasticity of the proteome of C. burnetii to battle tetracycline stress. Finally, we raise several plausible hypotheses that could further lead to more focused experiments on studying tetracycline resistance in C. burnetii and thus reduced treatment failures of Q fever

Assessment of Animal Sensitivity to Particularly Dangerous Orthopoxviruses, Using Primary Cultures of Lung Cells

Objective of the study is to investigate the sensitivity of different animals to highly pathogenic Orthopoxviruses applying techniques, based on utilization of primary cultures of lung cells, and to assess the possibility of further deployment of this approach. Materials and methods. Cultural and virological research methods are used. Results and conclusions. Performed is the assessment of sensitivity of outbred mice, marmots and chickens to variola virus (VV) and monkeypox virus (MPV), using suspended primary cultures of lung cells (SPCLC) of these animals. Through inoculation of the mentioned above cell cultures with VV and MPV in a dose of 0.00001 PFU per a cell (plaque forming unit /cell) demonstrated has been virus replication with maximum concentration values in all cases (1,4 - 2,0 lg PFU/ml), mainly 3 days after infection. According to the data on SPCLC, sensitivity to VV in mice, marmots and chickens (ID50 - 50 % infective dose) amounts to (1,3 ± 0,5) lg PFU; (2,3 ± 0,5) lg PFU; and (0,0 ± 0,4) lg PFU respectively, taking into account unhindered interaction of the virus with permissive lung cells in the organism of the animals. As for MPV values for this indicator, they are: (1,7 ± 0,3) lg PFU for mice, and (0,5 ± 0,3) lg PFU - for marmots. Obtained ID50 values for VV using mice SPCLC and for MPV using mice and marmots SPCLC coincide with the ones, studied in direct experiments on intranasal infection with the viruses, with regard to 10 % of the viral application in lungs when deploying the latter method of infection. The fact testifies to the possibility of further deployment of this method for the assessment of animal sensitivity to highly pathogenic Orthopoxviruses based on the results of in vitro experiments

Possibility of using a mouse SCID as a model animal to variola virus for evaluating anti-smallpox drug efficacy

At present, there is no animal model for smallpox that reflects the weakened immune system in people and can therefore help assess the prophylactic (highly preventive) efficiency of antiviral drugs. To fill in the gap, we have explored the possibility of using outbred immunodeficient SCID mice as a model animal for smallpox with the aid of virolo­gical, histological and electron microscopic and sta­tistical methods. There was no clinical evidence of disease by intranasal infection of mice at a dose of 5.2 log10 PFU (plaque forming units). At the same time, the 50 % infective dose (ID50) of VARV estimated for animals by registering the presence of the virus in their lungs after 4 days post i.n. infection was 3.5 log10 PFU and was relatively similar to that in humans, theoretically determined by identification of the clinical picture of the disease. Virus replication was detected only in the respiratory organs of mice challenged i.n. with VARV at a dose of 5.2 log10 PFU (50 ID50). The values for its concentrations in the lungs and nose resembled those for affected people and well-known animal models (Macaca cynomolgus and ICR mice), respiratorily infected with VARV at similar doses. The existing model animals were not significantly different from SCID mice in the duration of viral presence in the lungs. Moreover, in SCID mice, as in humans and other animal models, similar pathomor- phological changes of inflammatory necrotic nature in the respiratory organs have been reported. Using SCID mice in assessing the prophylactic efficacy of the antiviral drugs NIOCH-14 and ST-246 demonstrated the adequacy of the results obtained to those described in the literature. This opens up the prospect of using SCID mice as an animal model for smallpox to develop antiviral drugs intended for people with severe immuno­suppressive states

Non-alcoholic fatty liver disease and gastroesophageal reflux disease as a model of a patient with a high cardiovascular risk

A high level of cardiovascular diseases and the increase in the number of patients with excessive body weight indicate eating disorders among the population not only of Russia, but also in the whole world. Rhetorical remains the question of determining the primary pathogenetic link. However, the understanding and control of cardiovascular risk will entail improving the quality of life of patients with multiple organ pathology and positive changes in the dynamics of statistical indicators of health. The aim of our study was to study the features of cardiovascular system in 20 patients with nonalcoholic fatty liver disease (NAFLD) and gastroesophageal reflux disease (GERD) aged 18 to 60 years. As a result, we demonstrated a high prevalence of modifiable and non-modifiable factors of cardiovascular risk in patients with NAFLD and GERD, the leading of which was an inherited family history, arterial hypertension, inadequate physical activity, nicotine dependence, excessive alcohol consumption.Высокий уровень сердечно-сосудистых заболеваний и увеличение численности пациентов с избыточной массой тела свидетельствуют о нарушениях пищевого поведения среди населения не только россии, но и в мире в целом. риторическим остается вопрос определения первичного патогенетического звена. однако, понимание и возможность управления кардиоваскулярными рисками, повлечет за собой повышение качества жизни пациентов с полиорганной патологией и положительные сдвиги в динамике статистических показателей здравоохранения. Целью нашего исследования было изучение особенностей состояния сердечно-сосудистой системы у 20 больных с неалкогольной жировой болезнью печени (НаЖБП) и гастроэзофагеальной рефлюксной болезнью (ГЭрБ) в возрасте от 18 до 60 лет. В результате, нами была продемонстрирована высокая распространенность модифицируемых и немодифицируемых факторов кардиоваскулярных рисков у больных НаЖБП и ГЭрБ, ведущими из которых были наследственная отягощенность, артериальная гипертензия, недостаточная физическая активность, никотиновая зависимость, избыточное употребление алкоголя

Detection of the Genetic Material of the Viruses Tacheng uukuvirus and Sara tick phlebovirus in Taiga Ticks Collected in the Sverdlovsk, Tomsk Regions and Primorsky Territory of Russia and Their Phylogeny

Extensive spread of tick-borne diseases poses a significant problem for public health and the health of the population living in endemic areas.The aim of the study was to search, analyze genetic material and identify new viral agents of the Phenuiviridae family in taiga ticks collected in Asian regions of Russia using the method of high throughput sequencing.Materials and methods. The study involved 1460 taiga ticks collected in suburban areas of the Tomsk, Yekaterinburg and Primorsky Territory. The genetic material isolated from ticks was sequenced using Illumina technology followed by phylogenetic analysis.Results and discussion. Analysis of the sequencing results made it possible to detect extended nucleotide sequences of the L-gene fragment characteristic of the Phenuiviridae family viruses. We were able to identify 20 nucleotide sequences the length of 250 bp on average in homogenates of Ixodes persulcatus ticks. Eighteen isolates have been identified as members of the genus Uukuvirus and two isolates have been assigned to the genus Phlebovirus, Phenuiviridae family. Phylogenetic analysis has shown that all isolates of the genus Uukuvirus fall under the cluster of Tacheng tick virus 2 belonging to the species Tacheng uukuvirus. They form a separate phylogenetic group which is closely related to two Romanian variants of 2019. Tacheng tick virus 2 was detected in all three surveyed regions of the Asian part of Russia. Two Tomsk isolates of phlebovirus were classified as Sara tick phlebovirus and they clustered with two isolates of phleboviruses from Karelia. Thus, the genetic material of Tacheng tick virus 2 and Sara tick phlebovirus belonging to two genera of the family Phenuiviridae was found in I. persulcatus ticks collected in three geographically different regions of the Asian part of Russia

The Radio Spectral Index of the Crab Nebula

We present the results of a new, comprehensive investigation of the radio spectral index of the Crab Nebula supernova remnant. New data at 74 MHz are combined with data at 327 MHz, 1.5 GHz and 5 GHz. In contrast to previous claims, little spatial variation in the spectral index is seen. In particular, between 327 MHz and 5 GHz we see no evidence of spectral steepening near the edge of the nebula, the ``jet'' or the ionized filaments. The rms limits on any spectral index variations in these regions amount to no more than 0.01. We believe that earlier reports of large steepening were the result of correlator bias and image registration problems. An elongated feature was detected 1\arcmin northwest of the pulsar which may be a continuation of the well-known wisp-like structures seen closer to the center of the nebula. At 74 MHz, we see for the first time evidence of free-free absorption by the thermal material in the Crab Nebula's filaments. Apart from some possible renewed acceleration occurring in the wisps, the dominant accelerator of relativistic electrons in the Crab Nebula is the pulsar itself.Comment: 20 pages, Latex, requires aaspp4.st

Management and Performance of Diagnostic Investigations on the Platform of the Specialized Anti-Epidemic Team Mobile Complex During EVD Epidemics in 2014 in the Republic of Guinea

Given is an account of the EVD epidemics that started in the Republic of Guinea in December, 2013 and spread over West African countries within 2014. Established have been the grounds for the Rospotrebnadzor SAET deployment in the Republic of Guinea, objectives, goals and stuffing of the mission, and mobile complex technical performance. Described are the key stages of the work, including the process of integration into the UNMEER. Outlined are priority areas of collaboration with the National Public Health Ministry and international partner organizations. Represented are the results of work on the Ebola fever and other dangerous infectious diseases diagnostics, carried out at the mobile facility. Provided is molecular-genetic characteristics of the Ebola virus