Produção científica acerca do descolamento prematuro da placenta / Scientific production on abruptio placentae

Objetivo: analisar as produções científicas disponíveis na literatura, sobre descolamento prematuro da placenta (DPP). Métodos: trata-se de uma revisão integrativa de literatura. A busca foi realizada nas bases de dados Literatura Latino-Americana e do Caribe em Ciências da Saúde (LILACS) e MEDLINE pela Pubmed, a partir da questão norteadora “Quais as evidências científicas sobre descolamento prematuro da placenta?”. Resultados: predominaram estudos que não apresentam fortes evidências para aplicação clínica e a totalidade é de autoria médica. Os estudos analisados têm como foco a associação de DPP com patologias, alterações genéticas e comportamentais na gestação. Considerações finais: percebe-se a escassez com níveis de evidência altos, bem como, ausência de produções na área de enfermagem.

Objetive: this study aimed to analyze the scientific publications available in the literature onabruptio placentae. Methods: this is an integrative literature review. The search was conducted inthe databases Latin American and Caribbean Health Sciences (LILACS) and MEDLINE by PubMed, fromthe guiding question "What are the scientific evidence of abruptio placentae?”. Results: studies thatdo not show strong evidence for clinical application predominated, most medical authorship. Thestudies analysis were focused on the DPP association with diseases, genetic and behavioral changesduring pregnancy. Final Considerations: there is a shortage with high levels of evidence, as well asthe absence of productions in nursing.

Single point mutations in the helicase domain of the NS3 protein enhance dengue virus replicative capacity in human monocyte-derived dendritic cells and circumvent the type I interferon response.

Dengue is the most prevalent arboviral disease worldwide. The outcome of the infection is determined by the interplay of viral and host factors. In the present study, we evaluated the cellular response of human monocyte-derived DCs (mdDCs) infected with recombinant dengue virus type 1 (DV1) strains carrying a single point mutation in the NS3hel protein (L435S or L480S). Both mutated viruses infect and replicate more efficiently and produce more viral progeny in infected mdDCs compared with the parental, non-mutated virus (vBACDV1). Additionally, global gene expression analysis using cDNA microarrays revealed that the mutated DVs induce the up-regulation of the interferon (IFN) signalling and pattern recognition receptor (PRR) canonical pathways in mdDCs. Pronounced production of type I IFN were detected specifically in mdDCs infected with DV1-NS3hel-mutated virus compared with mdDCs infected with the parental virus. In addition, we showed that the type I IFN produced by mdDCs is able to reduce DV1 infection rates, suggesting that cytokine function is effective but not sufficient to mediate viral clearance of DV1-NS3hel-mutated strains. Our results demonstrate that single point mutations in subdomain 2 have important implications for adenosine triphosphatase (ATPase) activity of DV1-NS3hel. Although a direct functional connection between the increased ATPase activity and viral replication still requires further studies, these mutations speed up viral RNA replication and are sufficient to enhance viral replicative capacity in human primary cell infection and circumvent type I IFN activity. This information may have particular relevance for attenuated vaccine protocols designed for DV.

The impact of oocyte maturation media on early bovine embryonic development.

Successful production of high quality blastocysts in vitro depends on the use of a culture system that ensures the acquisition of developmental competence by the maturing oocyte. It is now clear that the in vitro maturation environment has a major influence on the oocyte's ability to acquire the potential to develop into blastocysts. In this work we examine the impact of oocyte culture media on the quality of blastocysts by comparing developmental rates, cell number and their allocation to embryonic cell lineages, apoptosis, and expression of developmentally important genes. Higher total cell count and ICM:TCN ratio, which are indicative of embryo viability, were observed in embryos derived from oocyte maturation in TCM-199 supplemented with serum when compared to blastocysts derived from oocyte maturation in SOF BSA. Moreover, oocyte maturation in TCM-199 supplemented with serum-generated embryos of higher morphological quality and producing higher levels of Interferon Tau transcripts when compared to embryos derived from oocyte maturation in SOF BSA. In conclusion, the oocyte maturation regimen affected the morphological feature of blastocysts, including total cell count and allocation of cells to trophectoderm (TE) and inner cell mass (ICM) lineages and the expression profiles of genes involved in various embryo functions such as early embryonic growth, regulation of gene transcription, trophoblast differentiation and function, embryo-maternal communication, and stress response. Our results show that the oocyte culture media have strong impact on the quality of embryos produced in vitro and emphasize the need for more in depth evaluation of oocyte maturation protocols.

Detection and characterization of rabies virus in Southern Brazil by PCR amplification and sequencing of the nucleoprotein gene.

Due to the medical and socio-economical importance of both human and animal rabies infection, several studies have suggested the use of molecular techniques such as RT-PCR and DNA sequencing for diagnosis and phylogenetic studies of the rabies virus. Considering the conservancy of the nucleoprotein (N) gene of the virus, we herein describe a RT-PCR assay for rabies diagnosis and characterization. A total of 75 samples obtained from a variety of animal species in the state of Santa Catarina (SC), Southern Brazil, were comparatively studied by fluorescence antibody test (FAT), mouse inoculation test (MIT), cell infection assay and RT-PCR, which revealed itself to be as sensitive as FAT and MIT and less time-consuming than MIT. Direct sequencing of the 5' end of the N gene allowed the clustering of the SC samples with samples from the vampire bat-related or sylvatic cycle through comparative sequence analysis.

Flow cytometry assay for intracellular rabies virus detection

Following previous studies reporting microbiological diagnosis by flow cytometry, the possibility of using this method was examined to monitor infection of susceptible cell lines by a fixed rabies virus strain (Pasteur Virus strain—PV) or a wild rabies virus strain (WRS). Suspensions of BHK-21 and C6 cells were infected with viruses and a time course of virus infection was established. Sequentially, at several time points, infected and control uninfected cells were fixed, permeabilized, and stained with a rabies virus-specific antibody conjugate. This was achieved by resuspending cells in a solution containing p-formaldehyde in FACS lysis fluid, which allowed the detection of intracellular virus with flourescein-coupled antibodies by flow cytometry. A Becton Dickinson FACSCalibur® flow cytometer was used to analyze the percentage of cells infected and the kinetics of the infection process was determined. As early as 12 h after inoculation with both rabies virus strains, significant levels (P<0.01) of infection (from 4.7 to 7.1%) were detected by flow cytometry. The maximum level of infection was obtained at 48 h in C6 cells (88%) with both viruses. The advantages of this method for examination of intracellular virus infection are discussed.

Isolation and replication of rabies virus in C6 rat glioma cells (clone CCL-107).

The susceptibility of the C6 rat glioma cell line (ATCC; CCL-107) to rabies virus was characterized. The kinetics of infection performed with a fixed and a wild strain (from an infected cow) of rabies virus was monitored by direct immunofluorescence. Fluorescent cytoplasmic bodies were readily observed by UV microscopy from 24 hours post-infection (hpi) onwards. The ability of C6 to produce rabies infective virion particles was confirmed by determining the viral titres present in the supernatants of infected cultures, by both BHK-21 cell infection and mice inoculation. C6 cells produced similar viral titres to those produced by BHK-21 for both strains used. In addition, the yield of rabies glycoprotein was assessed by ELISA. In general, BHK-21 and C6 cells infected either by PV or with the wild rabies strain produced similar amounts of rabies glycoprotein. At 96 hpi, however, when the glycoprotein production peaked, BHK-21 infected with the wild strain produced significantly higher amounts of glycoprotein than C6. Subsequently, the optimal conditions for isolation of wild rabies virus strains from C6 cells were established and these proved to be as sensitive as NA cells in detecting 10 wild rabies samples. Due to the high sensitivity exhibited, C6 rat glioma cells present a new and useful system for rabies virus investigation.

Isolation and replication of rabies virus in C6 rat glioma cells (clone CCL-107)

The susceptibility of the C6 rat glioma line (ATCC; CCL-107) to rabies virus was characterized. The kinetics of infection performed with a fixed and a wild strain (from an infected cow) of rabies virus was monitored by direct immunofluorescence. Fluorescent cytoplasmic bodies were readily observed by UV microscopy from 24 hours post-infection (hpi) onwards. The ability of C6 to produce rabies infective virion particles was confirmed by determining the viral titres present in the supertants of infected cultures by both BHK-21 cell infection and mice inoculation C6 cells produced similar viral titres to those produced by BHK-21 for both strains used. In addition, the yield of rabies glycoprotein was assessed by ELISA. In general, BHK-21 and C6 cells infected either by PV or with the wild rabies strain produced similar amounts of rabies glycoprotein. At 96 hpi, however, when the glycoprotein production peaked, BHK-21 infected with the wild strain produced significantly higher amounts of glycoprotein than C6. Subsequently, the optimal conditions for isolation of wild rabies virus strains from C6 cells were established and these proved to be as sensitive as NA cells in detecting 10 wild rabies samples. Due to the high sensitivity exhibited, C6 rat glioma cells present a new useful system for rabies virus investigation.