Эпидемиологические и вирусологические аспекты гриппа, острых респираторных вирусных инфекций и тяжелых острых респираторных инфекций в Республике Молдова, сезон 2018/2019

Agenția Națională pentru Sănătate Publică, Congresul al VIII-lea al specialiștilor din domeniul sănătății publice și managementului sanitar 24-25 octombrie 2019 Chișinău, Republica MoldovaSezonul de gripă 2018/2019 a debutat mai devreme în comparație cu sezoanele precedente. Perioada epidemică a debutat în săptămâna 03/2019, atingând apogeul în săptămâna 06/2019 și s-a terminat în săptămâna 11/2019. Virusul gripal A(H1)pdm09 a fost predominant, fi ind detectat în 79,8% din cazurile de gripă confi rmate de laborator. Virusul gripal A(H3) a fost detectat în 20,2% cazuri. Virusurile gripale de tip B nu au fost detectate în acest sezon. Toate (4) virusurile gripale A(H1)pdm09 caracterizate genetic aparțin subgrupei 6B.1 (reprezentată prin A/Michigan/45/2015). Principalele caracteristici ale virusurilor din grupa 6B.1 sunt prezența substituțiilor aminoacizilor S84N, S162N (cu apariția unui nou loc potențial de glicozilare) și I216T în HA1. Virusurile gripale A/H3 detectate aparțin subclaselor 3C.2a1b și 3C.2a2. În ceea ce privește virusurile H3N2 circulante în prezent, caracterizarea lor antigenică prin reacția de hemaglutinoinhibare rămâne dificilă din cauza aglutinării instabile a globulelor roșii de cobai, curcan și umane. Nivelul morbidității prin gripă a sporit de 2,2 ori față de sezonul 2017/2018. Nivelul morbidității prin IACRS în sezonul 2018/2019 s-a majorat de 1,07 ori, iar prin SARI – de 1,17 ori față de sezonul precedent. Începând cu săptămâna 40/2018, virusurile respiratorii negripale au fost detectate în 37,7% (130/345) din probe colectate în timpul monitoringului în sistemul de tip santinelă. Au predominat rinovirusurile, adenovirusurile și virusul respirator sincițial.The influenza season started earlier that previous seasons. The epidemic period started at week 03/2019, peaked in week 06/2019 and ended in week 11/2019. Influenza A(H1)pdm09 was the predominant virus detected in 79,8% of ILI cases. Influenza A(H3) was detected in 20,2%. Influenza B viruses were not detected this season. All (4) of the A(H1)pdm09 viruses genetically characterized, belonged to the subgroup 6B.1 (represented by A/Michigan/45/2015). The main characteristics of viruses in the 6B.1 group are that the viruses carry the amino acid substitutions S84N, S162N (introducing a new potential glycosylation site) and I216T in HA1. Detected influenza A/ H3 viruses belonged to subclades 3C.2a1b and 3C.2a2. In the case of currently circulating H3N2 virus’s antigenic characterization continues to be difficult by HI assay due to variable agglutination of red blood cells from guinea pig, turkey and humans. The level of influenza morbidity increased 2,2 times compared to 2017/2018 season. The level of ARI morbidity in this season increased 1,07 times and the SARI recorded increased 1,17 times compared to the previous season. Since week 40/2018 other respiratory viruses were detected in 37,7% (130/345) of samples in sentinel surveillance. Rhinovirus, adenovirus and respiratory syncytial virus was detected in higher level during this season.Сезон гриппа 2018/2019 начался раньше, чем в предыдущие сезоны. Эпидемический период начался на неделе 03/2019, достигнув максимума на неделе 06/2019, и завершился на неделе 11/2019. Вирус гриппа A(H1)pdm09 был доминирующим и выявлен в 79,8% случаев гриппа. Вирус гриппа A(H3) был обнаружен в 20,2% случаев. Вирусы гриппа B не были выявлены в этом сезоне. Все (4) вирусы гриппа A(H1)pdm09б охарактеризованные генетически, относятся к подгруппе 6B.1 (представленную вирусом A/Michigan/45/2015). Основные характеристики вирусов из подгруппы 6B.1 – это наличие аминокислотных замен S84N, S162N (с появлением нового потенциального места гликозилирования) и I216T в HA1. Выявленные вирусы гриппа A/H3 относятся к подклассам 3C.2a1b и 3C.2a2. В случае циркулирующих в настоящее время H3N2, их антигенная характеристика с помощью реакции торможения гемаглютинации по-прежнему затруднена из за нестабильной агглютинации эритроцитов морской свинки, индейки и человека. Уровень заболеваемости гриппом повысился в 2,2 раза по сравнению с сезоном 2017/2018. Заболеваемость ОРВИ в сезоне 2018/2019 увеличилась в 1,07 раз по сравнению с предыдущим сезоном, а количество случаев ТОРИ увеличилось в 1,17 раз. Начиная с недели 40/2018 другие респираторные вирусы были выявлены в 37,7% (130/345) образцов, собранных во время дозорного надзора. Риновирусы, аденовирусы и респираторно-синцитиальные вирусы выявлялись в большом количестве в течении этого сезона

A Potent and Selective Inhibitor of Cdc42 GTPase

Cdc42, a member of the Rho family of GTPases, has been shown to play a role in cell adhesion, cytoskeletal arrangement, phagocytosis and cell motility and migration, in addition to a host of other diverse biological processes. The function of Rho-family GTPases in disease pathogenesis has been well established and identification of small, cell permeable molecules that selectively and reversibly regulate Rho GTPases is of high scientific and potentially therapeutic interest. There has been limited success in identifying inhibitors that specifically interact with small Rho family GTPases. The identified probe, ML141 (CID-2950007), is demonstrated to be a potent, selective and reversible non-competitive inhibitor of Cdc42 GTPase suitable for in vitro assays, with low micromolar potency and selectivity against other members of the Rho family of GTPases (Rac1, Rab2, Rab7). Given the highly complementary nature of the function of the Rho family GTPases, Cdc42 selective inhibitors such as those reported here should help untangle the roles of the proteins in this family

Analysis of subcellular metabolite levels of potato tubers (Solanum tuberosum) displaying alterations in cellular or extracellular sucrose metabolism

The expression of a heterologous invertase in potato tubers (Solanum tuberosum) in either the cytosol or apoplast leads to a decrease in total sucrose content and to an increase in glucose. Depending on the targeting of the enzyme different changes in phenotype and metabolism of the tubers occur: the cytosolic invertase expressing tubers show an increase in the glycolytic flux, accumulation of amino acids and organic acids, and the appearance of novel disaccharides; however, these changes are not observed when the enzyme is expressed in the apoplast [Roessner et al. (2001). Plant Cell, 13, 11-29]. The analysis of these lines raised several questions concerning the regulation of compartmentation of metabolites in potato tubers. In the current study we addressed these questions by performing comparative subcellular metabolite profiling. We demonstrate that: (i) hexoses accumulate in the vacuole independently of their site of production, but that the cytosolic invertase expression led to a strong increase in the cytosolic glucose concentration and decrease in cytosolic sucrose, whereas these effects were more moderate in the apoplastic expressors; (ii) three out of four of the novel compounds found in the cytosolic overexpressors accumulate in the same compartment; (iii) despite changes in absolute cellular content the subcellular distribution of amino acids was invariant in the invertase overexpressing tubers. These results are discussed in the context of current models of the compartmentation of primary metabolism in heterotrophic plant tissues

Identification of a small molecule yeast TORC1 inhibitor with a flow cytometry-based multiplex screen

TOR (target of rapamycin) is a serine/threonine kinase, evolutionarily conserved from yeast to human, which functions as a fundamental controller of cell growth. The moderate clinical benefit of rapamycin in mTOR-based therapy of many cancers favors the development of new TOR inhibitors. Here we report a high throughput flow cytometry multiplexed screen using five GFPtagged yeast clones that represent the readouts of four branches of the TORC1 signaling pathway in budding yeast. Each GFP-tagged clone was differentially color-coded and the GFP signal of each clone was measured simultaneously by flow cytometry, which allows rapid prioritization of compounds that likely act through direct modulation of TORC1 or proximal signaling components. A total of 255 compounds were confirmed in dose-response analysis to alter GFP expression in one or more clones. To validate the concept of the high throughput screen, we have characterized CID 3528206, a small molecule most likely to act on TORC1 as it alters GFP expression in all five GFP clones in an analogous manner to rapamycin. We have shown that CID 3528206 inhibited yeast cell growth, and that CID 3528206 inhibited TORC1 activity both in vitro and in vivo with EC50s of 150 nM and 3.9 μM, respectively. The results of microarray analysis and yeast GFP collection screen further support the notion that CID 3528206 and rapamycin modulate similar cellular pathways. Together, these results indicate that the HTS has identified a potentially useful small molecule for further development of TOR inhibitors

CACHE (Critical Assessment of Computational Hit-finding Experiments): A public–private partnership benchmarking initiative to enable the development of computational methods for hit-finding

One aspirational goal of computational chemistry is to predict potent and drug-like binders for any protein, such that only those that bind are synthesized. In this Roadmap, we describe the launch of Critical Assessment of Computational Hit-finding Experiments (CACHE), a public benchmarking project to compare and improve small-molecule hit-finding algorithms through cycles of prediction and experimental testing. Participants will predict small-molecule binders for new and biologically relevant protein targets representing different prediction scenarios. Predicted compounds will be tested rigorously in an experimental hub, and all predicted binders as well as all experimental screening data, including the chemical structures of experimentally tested compounds, will be made publicly available and not subject to any intellectual property restrictions. The ability of a range of computational approaches to find novel binders will be evaluated, compared and openly published. CACHE will launch three new benchmarking exercises every year. The outcomes will be better prediction methods, new small-molecule binders for target proteins of importance for fundamental biology or drug discovery and a major technological step towards achieving the goal of Target 2035, a global initiative to identify pharmacological probes for all human proteins. [Figure not available: see fulltext.

Tamoxifen mechanically deactivates hepatic stellate cells via the G protein-coupled estrogen receptor

Tamoxifen has been used for many years to target estrogen receptor signalling in breast cancer cells. Tamoxifen is also an agonist of the G protein-coupled estrogen receptor (GPER), a GPCR ubiquitously expressed in tissues that mediates the acute response to estrogens. Here we report that tamoxifen promotes mechanical quiescence in hepatic stellate cells (HSCs), stromal fibroblast-like cells whose activation triggers and perpetuates liver fibrosis in hepatocellular carcinomas. This mechanical deactivation is mediated by the GPER/RhoA/myosin axis and induces YAP deactivation. We report that tamoxifen decreases the levels of hypoxia-inducible factor-1 alpha (HIF-1α) and the synthesis of extracellular matrix proteins through a mechanical mechanism that involves actomyosin-dependent contractility and mechanosensing of tissue stiffness. Our results implicate GPER-mediated estrogen signalling in the mechanosensory-driven activation of HSCs and put forward estrogenic signalling as an option for mechanical reprogramming of myofibroblast-like cells in the tumour microenvironment. Tamoxifen, with half a century of safe clinical use, might lead this strategy of drug repositioning.Peer reviewe

Complementarity Between a Docking and a High-Throughput Screen in Discovering New Cruzain Inhibitors†

Virtual and high-throughput screens (HTS) should have complementary strengths and weaknesses, but studies that prospectively and comprehensively compare them are rare. We undertook a parallel docking and HTS screen of 197861 compounds against cruzain, a thiol protease target for Chagas disease, looking for reversible, competitive inhibitors. On workup, 99 % of the hits were eliminated as false positives, yielding 146 well-behaved, competitive ligands. These fell into five chemotypes: two were prioritized by scoring among the top 0.1 % of the docking-ranked library, two were prioritized by behavior in the HTS and by clustering, and one chemotype was prioritized by both approaches. Determination of an inhibitor/cruzain crystal structure and comparison of the high-scoring docking hits to experiment illuminated the origins of docking false-negatives and false-positives. Prioritizing molecules that are both predicted by docking and are HTS-active yields well-behaved molecules, relatively unobscured by the false-positives to which both techniques are individually prone

Unexplored therapeutic opportunities in the human genome

A large proportion of biomedical research and the development of therapeutics is focused on a small fraction of the human genome. In a strategic effort to map the knowledge gaps around proteins encoded by the human genome and to promote the exploration of currently understudied, but potentially druggable, proteins, the US National Institutes of Health launched the Illuminating the Druggable Genome (IDG) initiative in 2014. In this article, we discuss how the systematic collection and processing of a wide array of genomic, proteomic, chemical and disease-related resource data by the IDG Knowledge Management Center have enabled the development of evidence-based criteria for tracking the target development level (TDL) of human proteins, which indicates a substantial knowledge deficit for approximately one out of three proteins in the human proteome. We then present spotlights on the TDL categories as well as key drug target classes, including G protein-coupled receptors, protein kinases and ion channels, which illustrate the nature of the unexplored opportunities for biomedical research and therapeutic development. © 2018 Macmillan Publishers Limited, part of Springer Nature. All rights reserved

Erratum: Unexplored therapeutic opportunities in the human genome (Nature reviews. Drug discovery (2018) 17 5 (317-332))

This corrects the article DOI: 10.1038/nrd.2018.14

Natural products in modern life science

With a realistic threat against biodiversity in rain forests and in the sea, a sustainable use of natural products is becoming more and more important. Basic research directed against different organisms in Nature could reveal unexpected insights into fundamental biological mechanisms but also new pharmaceutical or biotechnological possibilities of more immediate use. Many different strategies have been used prospecting the biodiversity of Earth in the search for novel structure–activity relationships, which has resulted in important discoveries in drug development. However, we believe that the development of multidisciplinary incentives will be necessary for a future successful exploration of Nature. With this aim, one way would be a modernization and renewal of a venerable proven interdisciplinary science, Pharmacognosy, which represents an integrated way of studying biological systems. This has been demonstrated based on an explanatory model where the different parts of the model are explained by our ongoing research. Anti-inflammatory natural products have been discovered based on ethnopharmacological observations, marine sponges in cold water have resulted in substances with ecological impact, combinatory strategy of ecology and chemistry has revealed new insights into the biodiversity of fungi, in depth studies of cyclic peptides (cyclotides) has created new possibilities for engineering of bioactive peptides, development of new strategies using phylogeny and chemography has resulted in new possibilities for navigating chemical and biological space, and using bioinformatic tools for understanding of lateral gene transfer could provide potential drug targets. A multidisciplinary subject like Pharmacognosy, one of several scientific disciplines bridging biology and chemistry with medicine, has a strategic position for studies of complex scientific questions based on observations in Nature. Furthermore, natural product research based on intriguing scientific questions in Nature can be of value to increase the attraction for young students in modern life science