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Abstract: The recombinant proteins KMPII, TRYP, and LACK of Leishmania infantum were produced in baculovirus-infected Trichoplusia ni larvae and used to analyze the seroreactivity of 165 dog serum samples by the multiple-well ELISA technique (57 infected dogs with clinical signs, 46 naturally infected and 11 experimentally infected; and 108 non-infected dogs, 76 from non-endemic areas and 32 from endemic areas). Recombinant (r) KMPII was the most recognized antigen, as the majority of infected dogs seroreacted against it (0.75). This is the first report of seroreactivity against rTRYP (0.51) and rLACK (0.42) in L. infantum-infected dogs, since previous studies using recombinant TRYP and LACK proteins produced in prokaryotic systems failed to detect specific seroreactivity. All non-infected dogs were negative for rTRYP and rLACK, and only one of the 32 from endemic areas seroreacted against rKMPII. The results demonstrate that L. infantum-infected dogs develop humoral immunity against rKMPII, rTRYP, and rLACK antigens. There was substantial agreement between crude total L. infantum antigen (CTLA)-based ELISA and rKMPII ELISA (κ =0.664), although this was higher than that found between the CTLA-based ELISA and rTRYP (κ =0.427) or rLACK (κ =0.343) ELISA, which can be interpreted as fair and moderate agreement, respectively. Ninety-three percent of the infected dogs analyzed developed specific antibodies against at least one of these three recombinant antigens. When the three recombinant antigen-based ELISA techniques were evaluated in parallel, almost perfect agreement (κ =0.880) with CTLA-based ELISA was observed, with a specificity of 0.97 and a sensitivity of 0.93 in relation to CTLA-based ELISA. Further studies using purified recombinant antigens in a single-well test or individually, depending on the objective of the study, are warranted. [Copyright &y& Elsevier]

Objective To review clinical outcomes after preimplantation genetic screening. Most methods of embryo viability assessment involve morphologic evaluation at different preimplantation developmental stages. A weak association between blastocyst morphology and aneuploidy has been described, supporting the basis for preimplantation genetic screening (PGS) for assessment of embryo viability. The expected improvement in reproductive outcome rates has been reached with the application of microarrays based on comparative genomic hybridization (CGH) in clinical routine PGS. Design Review of published studies and own unpublished data. Setting University-affiliated private institution. Patient(s) IVF patients undergoing PGS at different stages. Intervention(s) PGS with polar body, cleavage-stage, and blastocyst biopsies. Main Outcome Measure(s) Aneuploidy, implantation, and pregnancy rates. Results The clinical outcome after analysis of all 24 chromosomes improved pregnancy and implantation rates for different indications to a higher degree than the previously available technology, fluorescence in situ hybridization (FISH), in which only a limited number of chromosomes could be analyzed. Conclusion(s) Most of the data regarding the controversy of day-3 biopsy come from FISH cycles, and the utility of day-3 biopsy with new array-CGH technology should be further evaluated through randomized controlled trials. The current trend is blastocyst biopsy with a fresh transfer or vitrification for transfer in a nonstimulated cycle. [ABSTRACT FROM AUTHOR]

Niemann-Pick type C1 (NPC1) receptor is an endosomal membrane protein that regulates intracellular cholesterol traffic. This protein has been shown to play an important role for several viruses. It has been reported that SARS-CoV-2 enters the cell through plasma membrane fusion and/or endosomal entry upon availability of proteases. However, the whole process is not fully understood yet and additional viral/host factors might be required for viral fusion and subsequent viral replication. Here, we report a novel interaction between the SARS-CoV-2 nucleoprotein (N) and the cholesterol transporter NPC1. Furthermore, we have found that some compounds reported to interact with NPC1, carbazole SC816 and sulfides SC198 and SC073, were able to reduce SARS-CoV-2 viral infection with a good selectivity index in human cell infection models. These findings suggest the importance of NPC1 for SARS-CoV-2 viral infection and a new possible potential therapeutic target to fight against COVID-19. [Display omitted] • SARS-CoV-2 nucleoprotein (N) interacts with NPC1 protein. • This interaction was shown by CO-IP and reverse pull-downs. • Small molecules known to target EBOV GP interaction with NPC1 were tested as inhibitors. • These compounds inhibited SARS-CoV-2 and other CoV infections. [ABSTRACT FROM AUTHOR]

Despite the efforts to develop new treatments against Ebola virus (EBOV) there is currently no antiviral drug licensed to treat patients with Ebola virus disease (EVD). Therefore, there is still an urgent need to find new drugs to fight against EBOV. In order to do this, a virtual screening was done on the druggable interaction between the EBOV glycoprotein (GP) and the host receptor NPC1 with a subsequent selection of compounds for further validation. This screening led to the identification of new small organic molecules with potent inhibitory action against EBOV infection using lentiviral EBOV-GP-pseudotype viruses. Moreover, some of these compounds have shown their ability to interfere with the intracellular cholesterol transport receptor NPC1 using an ELISA-based assay. These preliminary results pave the way to hit to lead optimization programs that lead to successful candidates. Image 1 • Actually there is no antiviral drugs to treat patients with Ebola virus disease. • The interaction between EBOV glycoprotein (GP) and the host receptor NPC1 is a well-known druggable target. • Virtual screening on this interaction allowed us the selection of candidates for experimental validation. • Identification of new small molecules able to inhibit EBOV infection using lentiviral EBOV-GP-pseudotype viruses. [ABSTRACT FROM AUTHOR]