In silico screening for human norovirus antivirals reveals a novel non-nucleoside inhibitor of the viral polymerase

Human norovirus causes approximately 219,000 deaths annually, yet there are currently no antivirals available. A virtual screening of commercially available drug-like compounds (~300,000) was performed on the suramin and PPNDS binding-sites of the norovirus RNA-dependent RNA polymerase (RdRp). Selected compounds (n = 62) were examined for inhibition of norovirus RdRp activity using an in vitro transcription assay. Eight candidates demonstrated RdRp inhibition (>25% inhibition at 10 μM), which was confirmed using a gel-shift RdRp assay for two of them. The two molecules were identified as initial hits and selected for structure-activity relationship studies, which resulted in the synthesis of novel compounds that were examined for inhibitory activity. Five compounds inhibited human norovirus RdRp activity (>50% at 10 μM), with the best candidate, 54, demonstrating an IC50 of 5.6 μM against the RdRp and a CC50 of 62.8 μM. Combinational treatment of 54 and the known RdRp site-B inhibitor PPNDS revealed antagonism, indicating that 54 binds in the same binding pocket. Two RdRps with mutations (Q414A and R419A) previously shown to be critical for the binding of site-B compounds had no effect on inhibition, suggesting 54 interacts with distinct site-B residues. This study revealed the novel scaffold 54 for further development as a norovirus antiviral

Eumalacostracan phylogeny and total evidence: limitations of the usual suspects

<p>Abstract</p> <p>Background</p> <p>The phylogeny of Eumalacostraca (Crustacea) remains elusive, despite over a century of interest. Recent morphological and molecular phylogenies appear highly incongruent, but this has not been assessed quantitatively. Moreover, 18S rRNA trees show striking branch length differences between species, accompanied by a conspicuous clustering of taxa with similar branch lengths. Surprisingly, previous research found no rate heterogeneity. Hitherto, no phylogenetic analysis of all major eumalacostracan taxa (orders) has either combined evidence from multiple loci, or combined molecular and morphological evidence.</p> <p>Results</p> <p>We combined evidence from four nuclear ribosomal and mitochondrial loci (18S rRNA, 28S rRNA, 16S rRNA, and cytochrome <it>c </it>oxidase subunit I) with a newly synthesized morphological dataset. We tested the homogeneity of data partitions, both in terms of character congruence and the topological congruence of inferred trees. We also performed Bayesian and parsimony analyses on separate and combined partitions, and tested the contribution of each partition. We tested for potential long-branch attraction (LBA) using taxon deletion experiments, and with relative rate tests. Additionally we searched for molecular polytomies (spurious clades). Lastly, we investigated the phylogenetic stability of taxa, and assessed their impact on inferred relationships over the whole tree. We detected significant conflict between data partitions, especially between morphology and molecules. We found significant rate heterogeneity between species for both the 18S rRNA and combined datasets, introducing the possibility of LBA. As a test case, we showed that LBA probably affected the position of Spelaeogriphacea in the combined molecular evidence analysis. We also demonstrated that several clades, including the previously reported and surprising clade of Amphipoda plus Spelaeogriphacea, are 'supported' by zero length branches. Furthermore we showed that different sets of taxa have the greatest impact upon the relationships within molecular versus morphological trees.</p> <p>Conclusion</p> <p>Rate heterogeneity and conflict between data partitions mean that existing molecular and morphological evidence is unable to resolve a well-supported eumalacostracan phylogeny. We believe that it will be necessary to look beyond the most commonly utilized sources of data (nuclear ribosomal and mitochondrial sequences) to obtain a robust tree in the future.</p

Expression of angiogenic regulators, VEGF and leptin, is regulated by the EGF/PI3K/STAT3 pathway in colorectal cancer cells.

Abstract Both leptin and vascular endothelial growth factor (VEGF) are growth and angiogenic cytokines that are upregulated in different types of cancer and have been implicated in neoplastic progression. Here we investigated the molecular mechanism by which leptin and VEGF expression are regulated in colon cancer by epidermal growth factor (EGF). In colon cancer cell line HT-29, EGF induced the binding of signal transducer and activator transcription 3 (STAT3) to STAT3 consensus motifs within the VEGF and leptin promoters and stimulated leptin and VEGF mRNA and protein synthesis. All these EGF effects were significantly blocked when HT-29 cells were treated with an inhibitor of the phosphoinositide 3-kinase (PI3K) pathway, LY294002, or with small interfering RNA (siRNA) targeting STAT3. Thus, our study identified the EGF/PI3K/STAT3 signaling as an essential pathway regulating VEGF and leptin expression in EGF-responsive colon cancer cells. This suggests that STAT3 pathways might constitute attractive pharmaceutical targets in colon cancer patients where anti-EGF receptor drugs are ineffective

CXCL12/SDF-1 from perisynaptic Schwann cells promotes regeneration of injured motor axonterminals

The neuromuscular junction has retained through evolution the capacity to regenerate after damage, but little is known on the inter-cellular signals involved in its functional recovery from trauma, autoimmune attacks, or neurotoxins. We report here that CXCL12, also abbreviated as stromal-derived factor-1 (SDF-1), is produced specifically by perisynaptic Schwann cells following motor axon terminal degeneration induced by -latrotoxin. CXCL12 acts via binding to the neuronal CXCR4 receptor. A CXCL12-neutralizing antibody or a specific CXCR4 inhibitor strongly delays recovery from motor neuron degeneration invivo. Recombinant CXCL12 invivo accelerates neurotransmission rescue upon damage and very effectively stimulates the axon growth of spinal cord motor neurons invitro. These findings indicate that the CXCL12-CXCR4 axis plays an important role in the regeneration of the neuromuscular junction after motor axon injury. The present results have important implications in the effort to find therapeutics and protocols to improve recovery of function after different forms of motor axon terminal damage

Acarinose da videira no Rio Grande do Sul.

bitstream/item/48495/1/Circular-Tecnica-85.pd

A new species of Armascirus and description of the male of Scutopalus tomentosus from Brazil (Acari: Cunaxidae).

In this work, we describe the male of Scutopalus tomentosus Rocha, Skvarla & Ferla, 2013 and a new speciesArmascirus amazoniensis Wurlitzer & Silvasp. nov. from specimens collected on coconut crop, Cocos nuciferaL. (Arecaceae), cultivated in state of Pará, into the Amazonic biome, Brazil

First insights into the microbiology of three antarctic briny systems of the northern Victoria land

Different polar environments (lakes and glaciers), also in Antarctica, encapsulate brine pools characterized by a unique combination of extreme conditions, mainly in terms of high salinity and low temperature. Since 2014, we have been focusing our attention on the microbiology of brine pockets from three lakes in the Northern Victoria Land (NVL), lying in the Tarn Flat (TF) and Boulder Clay (BC) areas. The microbial communities have been analyzed for community structure by next generation sequencing, extracellular enzyme activities, metabolic potentials, and microbial abundances. In this study, we aim at reconsidering all available data to analyze the influence exerted by environmental parameters on the community composition and activities. Additionally, the prediction of metabolic functions was attempted by the phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt2) tool, highlighting that prokaryotic communities were presumably involved in methane metabolism, aromatic compound biodegradation, and organic compound (proteins, polysaccharides, and phosphates) decomposition. The analyzed cryoenvironments were different in terms of prokaryotic diversity, abundance, and retrieved metabolic pathways. By the analysis of DNA sequences, common operational taxonomic units ranged from 2.2% to 22.0%. The bacterial community was dominated by Bacteroidetes. In both BC and TF brines, sequences of the most thermally tolerant and methanogenic Archaea were detected, some of them related to hyperthermophiles

Shallow water tomography in a highly variable scenario

In October 2000, SiPLAB and the Instituto Hidrografico (IH - PN) conducted the IN-TIFANTE'00 sea trial in a shallow area off the Peninsula of Troia, approximately 50 km south from Lisbon, in Portugal. The experiment itself and results obtained in most of the data set have been reported at various occasions in the last two years. This paper focuses on the data acquired during Event 2, where the acoustic propagation path was approximately range independent and the source ship was held on station at a constant range of 5.8 km from the vertical line array. Although these conditions were, in general, relatively benign for matched-field tomography, retrieval of water column and bottom parameters over a 14-hour-long recording revealed to be extremely difficult. This paper analysis in detail the characteristics of this data set and determines the causes for the observed inversion difficulties. Is is shown that the causes for the poor performance of the conventional methods are mainly the tide induced spatially correlated noise and the relative source-receiver motion during time averaging. An eigenvalue-based criterion is proposed for detecting optimal averaging time. It is shown that this data selection procedure together with hydrophone normalization and an appropriate objective function provide a better model fit and consistent inversion results and thus a better understanding of the environmental variability

Clindamycin phospate-zinc acetate versus clindamycin phosphate-zinc acetate + adapalene in the treatment of mild to moderate acne. Results of a multicentre, randomized, retrospective, sponsor-free study

Introduction: The aim of the present study was to evaluate the efficacy and the response in patients with mild to moderate acne of a clindamycin phosphate-zinc acetate topical therapy in comparison with clindamycin phosphate-zinc acetate plus adapalene. Methods: Patients with mild to moderate acne were randomized into two groups and treated, respectively, with a gel containing 1 % clindamycin phosphate-0.5% zinc acetate (2 applications/day for 12 weeks) or with the same gel (1 application/day for 12 weeks) plus a gel containing 0.1% adapalene (1 application/day for 12 weeks). No other topical or systemic drugs were allowed, except for a detergent and a sunscreen. Acne severity and treatment efficacy were evaluated by means of the Global Acne Grading System (GAGS). Results: At the end of the study, 63 patients were considered evaluable (29 patients in the group treated with clindamycin-zinc and 34 in the group treated with clindamycin-zinc and adapalene). Significant clinical improvement (maggiore/uguale 50% from baseline) was observed in 12/29 patients (41.4%) in the group treated with clindamycin-zinc and in 22/34 patients (64.7%) in the group treated with clindamycin-zinc and adapalene (p< 0.05). Irritant contact dermatitis was observed in 12 patients (3 in the group treated with clindamycin-zinc and 9 in the group treated with clindamycin-zinc and adapalene); in the latter group, two patients stopped the treatment. Conclusions: On the basis of the results of this study, which is the first one on the activity and tolerability of the association clindamycin-zinc, the latter association is less effective than the association clindamycin-zinc and adapalene