Peptidomimetic and Non- Peptidomimetic Derivatives as Possible SARS-CoV-2 Main Protease (Mpro) Inhibitors

To design novel inhibitors of the SARS-CoV-2 main protease (Mpro), we investigated the binding mode of the recently reported α-ketoamide inhibitors of this enzyme. Following, we utilized in-silico screening to identify 168 peptidomimetic and non-peptidomimetic compounds that are high probability Mpro binding candidates. The compounds were synthesized in 5 to 10 mg for initial screening for their potential inhibition of Mpro using Fluorescence Resonance Energy Transfer (FRET) assay. The study was conducted using the main protease, MBP-tagged (SARS-CoV-2) Assay Kit (BPS Bioscience, #79955-2), and the fluorescence due to enzymatic cleavage of substrate measured using BMG LABTECH CLARIOstar™, a fluorescent microplate reader, with an excited/emission wavelength of 360 nm/460 nm, respectively. The FRET assay showed 29 compounds to exhibit lower fluorescence compared to the positive control, indicating inhibitory activity, with three of the compounds exhibiting over 50% enzymatic inhibition. The assay average scores were plotted as dose inhibition curves using variable parameter nonlinear regression to calculate the IC50 values. To design more potent inhibitors, an in-silico molecular docking simulation using the SARS-CoV-2 Mpro crystal structure was conducted to investigate on a molecular level the key binding residues at the active site, as well as the possible binding modes and affinity of the lead inhibitors. Additionally, an in-silico study of the compounds\u27 molecular properties and physicochemical profiles was performed to predict their pharmacokinetic properties and assess their suitability as potential orally active drug candidates.https://scholarscompass.vcu.edu/gradposters/1139/thumbnail.jp

Эпидемиология и структура сочетанной черепно-мозговой и скелетной травмы в г. Ташкенте

In period of 2001—2004 in Tashkent we observe growing concomitant craniocerebral and skeletal injuries (CCCSI) on 4,8% per annum. People of working age (at the age of 21—60) often get concomitant craniocerebral and skeletal injuries (CCCSI) though males dominate at the age of 21—40. But at the age of 15—20 and above 60 years old females get a CCCSI in 2—2,5 times higher, than males.Patients admission to hospital at different times greatly differentiates: the largest was in period from 13 to 18 hours and smallest was from 0 to 6 hours. Road trauma appearance is principal cause of concomitant craniocerebral and skeletal injuries (CCCSI) in Tashkent in all the years of observation. It has the tendency to grow from 19,4 to 29,1%. Noticeable growing of concomitant craniocerebral and skeletal injuries (CCCSI) is alarming at the work and in sport. The severe skeleton injuries were dominant among all kind of injuries.В Ташкенте за период наблюдения с 2001 по 2004 г. отмечается рост сочетанной черепно-мозговой и скелетной травмы (СЧМСТ) на 4,8% в год. Чаще имели СЧМСТ люди трудоспособного возраста (21-60 лет) - 74,9%, причем мужчины доминировали в возрасте 21-40 лет (54,5%), а в возрасте 15-20 лет и старше 60 лет женщины (в 2-2,5 раза чаще мужчин). Поступление пострадавших по времени суток существенно отличается: наибольшее в период с 13 до 18 ч и наименьшее с 0 до 6 ч. Автодорожная травма остается основной причиной возникновения СЧМСТ в Ташкенте во все годы наблюдения, имея тенденцию к росту от 19,4 до 29,1%. Тревожным следует считать заметный рост СЧМСТ на производстве и в спортивных учреждениях. Во всех видах травм чаще всего возникли ведущие тяжелые скелетные травмы

Drosophila selenophosphate synthetase 1 regulates vitamin B6 metabolism: prediction and confirmation

<p>Abstract</p> <p>Background</p> <p>There are two selenophosphate synthetases (SPSs) in higher eukaryotes, SPS1 and SPS2. Of these two isotypes, only SPS2 catalyzes selenophosphate synthesis. Although SPS1 does not contain selenophosphate synthesis activity, it was found to be essential for cell growth and embryogenesis in <it>Drosophila</it>. The function of SPS1, however, has not been elucidated.</p> <p>Results</p> <p>Differentially expressed genes in <it>Drosophila </it>SL2 cells were identified using two-way analysis of variance methods and clustered according to their temporal expression pattern. Gene ontology analysis was performed against differentially expressed genes and gene ontology terms related to vitamin B6 biosynthesis were found to be significantly affected at the early stage at which megamitochondria were not formed (day 3) after <it>SPS1 </it>knockdown. Interestingly, genes related to defense and amino acid metabolism were affected at a later stage (day 5) following knockdown. Levels of pyridoxal phosphate, an active form of vitamin B6, were decreased by <it>SPS1 </it>knockdown. Treatment of SL2 cells with an inhibitor of pyridoxal phosphate synthesis resulted in both a similar pattern of expression as that found by <it>SPS1 </it>knockdown and the formation of megamitochondria, the major phenotypic change observed by <it>SPS1 </it>knockdown.</p> <p>Conclusions</p> <p>These results indicate that SPS1 regulates vitamin B6 synthesis, which in turn impacts various cellular systems such as amino acid metabolism, defense and other important metabolic activities.</p

Иммуноопосредованная энцефалопатия на фоне эффективной терапии аденокарциномы легкого анти-PD-1. Клиническое наблюдение

Now the number of patients receiving immunotherapy with checkpoint inhibitors, including nivolumab, is growing all over the world. At the same time, there is a growing of immune-related adverse events in clinical practice, including rare ones. There is a case report of immune-related encephalopathy (grade 3) at patient with metastatic non-small cell lung cancer after complete response at nivolumab treatment.В настоящий момент во всем мире растет число пациентов, получающих иммунотерапию ингибиторами контрольных точек, включая ниволумаб. Вместе с тем растет и вероятность встретиться в клинической практике с иммуноопосредованными нежелательными явлениями, в том числе достаточно редкими. Приводится клиническое наблюдение высокоэффективного лечения ниволумабом больной метастатическим немелкоклеточным раком легкого с полным клиническим и рентгенологическим эффектом, осложнившегося иммуноопосредованной энцефалопатией 3 степени

Exchange of functional domains between a bacterial conjugative relaxase and the integrase of the human adeno-associated virus

Endonucleases of the HUH family are specialized in processing single-stranded DNA in a variety of evolutionarily highly conserved biological processes related to mobile genetic elements. They share a structurally defined catalytic domain for site-specific nicking and strand-transfer reactions, which is often linked to the activities of additional functional domains, contributing to their overall versatility. To assess if these HUH domains could be interchanged, we created a chimeric protein from two distantly related HUH endonucleases, containing the N-terminal HUH domain of the bacterial conjugative relaxase TrwC and the C-terminal DNA helicase domain of the human adeno-associated virus (AAV) replicase and site-specific integrase. The purified chimeric protein retained oligomerization properties and DNA helicase activities similar to Rep68, while its DNA binding specificity and cleaving-joining activity at oriT was similar to TrwC. Interestingly, the chimeric protein could catalyse site-specific integration in bacteria with an efficiency comparable to that of TrwC, while the HUH domain of TrwC alone was unable to catalyze this reaction, implying that the Rep68 C-terminal helicase domain is complementing the TrwC HUH domain to achieve site-specific integration into TrwC targets in bacteria. Our results illustrate how HUH domains could have acquired through evolution other domains in order to attain new roles, contributing to the functional flexibility observed in this protein superfamily.This work was supported by the Medical Research Council (MRC) grant MR/N022890/1 to EH and grant 1001764 to RML; National Institutes of Health (NIH) grant RO1-GM09285 to CRE; Spanish Ministry of Economy and competitiveness (MINECO) grant BIO2013-46414-P to ML and AFM is supported by a Doc.Mobility fellowship from the Swiss National Science Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Clinical and Prognostic Significance of Heart Rate Variability in Pulmonary Hypertension

34th Annual Meeting and Scientific Sessions of the International-Society-for-Heart-and-Lung-Transplantation -- APR 10-13, 2014 -- San Diego, CAWOS: 000333866700620Int Soc Heart & Lung Transplanta