New nicotinic acid-based 3,5-diphenylpyrazoles: design, synthesis and antihyperlipidemic activity with potential NPC1L1 inhibitory activity.

Nicotinic acid hydrazide was incorporated into new 4,5-dihydro-5-hydroxy-3,5-diphenylpyrazol-1-yl derivatives. Compounds 6a-h were synthesized, and their antihyperlipidemic activity was evaluated in high cholesterol diet-fed rat model. Compounds 6e, 6f were found to decrease the levels of serum total cholesterol by 14-19% compared to control group. Total triglycerides were also reduced by 24-28% and LDL cholesterol by 16%. As expected from parent niacin, compounds 6e and 6f caused an elevation of HDL cholesterol by 33-41%. Docking study supported the ability of designed compounds to block NPC1L1 active site in a manner similar to that observed with ezetimibe.

l-Carnitine ameliorates the osteoporotic changes and protects against simvastatin induced myotoxicity and hepatotoxicity in glucocorticoid-induced osteoporosis in rats.

The current study aimed to discover more effective drugs to treat osteoporosis (OP) with fewer side effects. OP was induced in 24 rats using dexamethasone (DEX) 7 mg/kg intramuscular once weekly for four weeks, with six rats as a negative control. The osteoporotic rats were divided into one untreated group (positive control) and three treated groups (n = 6) that received L-carnitine (L-Car) (100 mg/kg/d), simvastatin (SIMV) (10 mg/kg/d), and L-Car + SIMV in the same previous doses, all treatments were orally for four weeks. At the end of the experiment, serum calcium (Ca), phosphorous (P), alkaline phosphatase (ALP), osteoprotegerin (OPG), total antioxidant (TAO), creatine kinase (CK), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) levels were measured. The femur was histopathologically examined. Serum Ca, OPG, and TAO levels increased significantly, while P and ALP levels decreased in the L-Car and SIMV treated groups compared to the DEX-treated group. Moreover, there was a significant decrease in CK, ALT, and AST levels in the L-Car and L-Car + SIMV treated groups compared to the DEX treated group. CONCLUSIONS: L-Car and SIMV have antiosteoporotic effects, as well as a synergistic effect. Moreover, L-Car ameliorates SIMV-induced myotoxicity and hepatoxicity.

Thieno[2,3-b]thiophene Derivatives as Potential EGFRWT and EGFRT790M Inhibitors with Antioxidant Activities: Microwave-Assisted Synthesis and Quantitative In Vitro and In Silico Studies.

Microwave-assisted synthesis and spectral analysis of certain novel derivatives of 3,4-diaminothieno[2,3-b]thiophene-2,5-dicarbonitrile 1-7 were carried out. Compounds 1-7 were examined for cytotoxicity against MCF-7 and A549 cell lines using the quantitative MTT method, and gefitinib and erlotinib were used as reference standards. Compounds 1-7 were shown to be more active than erlotinib against the two cell lines tested. Compound 2 outperformed regular erlotinib by 4.42- and 4.12-fold in MCF-7 and A549 cells, respectively. The most cytotoxic compounds were subsequently studied for their suppression of kinase activity using the homogeneous time-resolved fluorescence assay versus epidermal growth factor receptor (EGFRWT) and EGFR790M. With IC50 values of 0.28 ± 0.03 and 5.02 ± 0.19, compound 2 was demonstrated to be the most effective against both forms of EGFR. Furthermore, compound 2 also had the best antioxidant property, decreasing the radical scavenging activity by 78%. Molecular docking research, on the other hand, was carried out for the analyzed candidates (1-7) to study their mechanism of action as EGFR inhibitors. In silico absorption, distribution, metabolism, excretion, and toxicity tests were also performed to explain the physicochemical features of the examined derivatives.

Carvedilol induces the antiapoptotic proteins Nrf2 and Bcl2 and inhibits cellular apoptosis in aluminum-induced testicular toxicity in male Wistar rats.

The present study was carried out to explore the protective effect of carvedilol (CARV) on aluminum chloride-induced testicular damage in Westar rats. Forty adult male rats, aged 8 weeks, were randomly divided into 4 groups (10 rats each). Group I (control group) received normal saline; whereas group II animals were supplemented with CARV in a dose of 10 mg/kg/day. Group III received AlCl3 (30 mg/kg/day) whereas group IV was co-administered CARV and AlCl3 as the same doses in group II and III respectively. The route of the application was oral gavage for CARV and I.P for AlCl3 for 20 successive days. Exposure of rats to AlCl3 for 20 consecutive days resulted in a significant decrease in serum and testicular superoxide dismutase and catalase activities, serum testosterone level, and sperm count and motility; on the other hand, an increase in nitric oxide, malondialdehyde, aluminum, and serum tumor necrosis factor-alpha levels. Furthermore, histopathological changes in the testis exhibited marked testicular damage. In addition, it revealed a significant up-regulation in the level of the expression for the apoptotic marker; Caspase-3, and down-regulation in antiapoptotic marker Bcl2 and Nrf2 genes. On the other hand, the co-administration of CARV modulated the biochemical parameters, saved sperm count and motility, and the histopathological findings, also, restored the observed changes in Caspase-3, Bcl2, and Nrf2 transcriptional genes. These data suggested that administration of CARV protects against AlCl3 induced testicular oxidative, inflammatory, and apoptosis damage.

Standardized phylogenetic and molecular evolutionary analysis applied to species across the microbial tree of life.

There is growing interest in reconstructing phylogenies from the copious amounts of genome sequencing projects that target related viral, bacterial or eukaryotic organisms. To facilitate the construction of standardized and robust phylogenies for disparate types of projects, we have developed a complete bioinformatic workflow, with a web-based component to perform phylogenetic and molecular evolutionary (PhaME) analysis from sequencing reads, draft assemblies or completed genomes of closely related organisms. Furthermore, the ability to incorporate raw data, including some metagenomic samples containing a target organism (e.g. from clinical samples with suspected infectious agents), shows promise for the rapid phylogenetic characterization of organisms within complex samples without the need for prior assembly.

Phylogenetic analysis of the NEEP21/calcyon/P19 family of endocytic proteins: evidence for functional evolution in the vertebrate CNS.

Endocytosis and vesicle trafficking are required for optimal neural transmission. Yet, little is currently known about the evolution of neuronal proteins regulating these processes. Here, we report the first phylogenetic study of NEEP21, calcyon, and P19, a family of neuronal proteins implicated in synaptic receptor endocytosis and recycling, as well as in membrane protein trafficking in the somatodendritic and axonal compartments of differentiated neurons. Database searches identified orthologs for P19 and NEEP21 in bony fish, but not urochordate or invertebrate phyla. Calcyon orthologs were only retrieved from mammalian databases and distant relatives from teleost fish. In situ localization of the P19 zebrafish ortholog, and extant progenitor of the gene family, revealed a CNS specific expression pattern. Based on non-synonymous nucleotide substitution rates, the calcyon genes appear to be under less intense negative selective pressure. Indeed, a functional group II WW domain binding motif was detected in primate and human calcyon, but not in non-primate orthologs. Sequencing of the calcyon gene from 80 human subjects revealed a non-synonymous single nucleotide polymorphism that abrogated group II WW domain protein binding. Altogether, our data indicate the NEEP21/calcyon/P19 gene family emerged, and underwent two rounds of gene duplication relatively late in metazoan evolution (but early in vertebrate evolution at the latest). As functional studies suggest NEEP21 and calcyon play related, but distinct roles in regulating vesicle trafficking at synapses, and in neurons in general, we propose the family arose in chordates to support a more diverse range of synaptic and behavioral responses.

Draft Genome Sequence of Thauera sp. Strain SWB20, Isolated from a Singapore Wastewater Treatment Facility Using Gel Microdroplets.

We report here the genome sequence of Thauera sp. strain SWB20, isolated from a Singaporean wastewater treatment facility using gel microdroplets (GMDs) and single-cell genomics (SCG). This approach provided a single clonal microcolony that was sufficient to obtain a 4.9-Mbp genome assembly of an ecologically relevant Thauera species.

Genomic comparison of Escherichia coli O104:H4 isolates from 2009 and 2011 reveals plasmid, and prophage heterogeneity, including shiga toxin encoding phage stx2.

In May of 2011, an enteroaggregative Escherichia coli O104:H4 strain that had acquired a Shiga toxin 2-converting phage caused a large outbreak of bloody diarrhea in Europe which was notable for its high prevalence of hemolytic uremic syndrome cases. Several studies have described the genomic inventory and phylogenies of strains associated with the outbreak and a collection of historical E. coli O104:H4 isolates using draft genome assemblies. We present the complete, closed genome sequences of an isolate from the 2011 outbreak (2011C-3493) and two isolates from cases of bloody diarrhea that occurred in the Republic of Georgia in 2009 (2009EL-2050 and 2009EL-2071). Comparative genome analysis indicates that, while the Georgian strains are the nearest neighbors to the 2011 outbreak isolates sequenced to date, structural and nucleotide-level differences are evident in the Stx2 phage genomes, the mer/tet antibiotic resistance island, and in the prophage and plasmid profiles of the strains, including a previously undescribed plasmid with homology to the pMT virulence plasmid of Yersinia pestis. In addition, multiphenotype analysis showed that 2009EL-2071 possessed higher resistance to polymyxin and membrane-disrupting agents. Finally, we show evidence by electron microscopy of the presence of a common phage morphotype among the European and Georgian strains and a second phage morphotype among the Georgian strains. The presence of at least two stx2 phage genotypes in host genetic backgrounds that may derive from a recent common ancestor of the 2011 outbreak isolates indicates that the emergence of stx2 phage-containing E. coli O104:H4 strains probably occurred more than once, or that the current outbreak isolates may be the result of a recent transfer of a new stx2 phage element into a pre-existing stx2-positive genetic background.