Validation of a DNA-Based Next-Generation Sequencing Test for Molecular Diagnostic Variant and Fusion Detection in Formalin-Fixed, Paraffin-Embedded Tissue Specimens and Liquid Biopsy Plasma/Cell-Free DNA Samples.

This study evaluated two DNA-based next-generation sequencing approaches for detection of single-nucleotide variants (SNVs) and fusions in formalin-fixed, paraffin-embedded (FFPE) tissue specimens and liquid biopsies (AVENIO Targeted and Surveillance Panels). Reference standards (n = 7 with SNVs and structural variants) and real-world FFPE tissue specimens (n = 26 lung, colorectal, pancreas, ovary, breast, prostate, melanoma, and soft tissue cancer cases with n = 27 samples), liquid biopsies [n = 29 cases with n = 40 plasma/cell-free DNA (cfDNA) samples], and one pleural effusion (lung cancer) were analyzed by the AVENIO workflow for known SNVs (BRAF, BRCA1/2, CTNNB1, EGFR, KRAS, MET exon 14 skipping, NRAS, PIK3CA, and TP53), insertions and deletions (ERBB2 and KIT), and fusions (ALK and ROS1). Detection of SNVs, insertions and deletions, and fusions was reliable in 24 of 26 FFPE tissue specimen cases and at 1% allele frequency in 5 of 5 cfDNA reference standards and 37 of 40 plasma/cfDNA samples. Pitfalls were identified for the AVENIO workflow in calling and listing of clinically relevant variants, requiring additional manual inspection. Moreover, laboratory workflows are distinct for FFPE tissue specimens and liquid biopsies as well as time-consuming for sample quality control assays. In summary, the DNA-based next-generation sequencing approaches may be suitable for routine molecular pathology diagnostics on careful data interpretation and further optimization of the technical and laboratory workflows.

Nutrição e doença de alzheimer no idoso: uma revisão / Nutrition and alzheimer's disease in older adults: a review

Objetivo: verificar se a alimentação e a nutrição podem contribuir para a prevenção ou retardo da progressão da doença de Alzheimer. Métodos: pesquisa de revisão integrativa cujos artigos foram encontrados nas bases de dados: Lilacs, Medline, Periódicos Capes e National Center of Biotechnology Information, entre o período de 2006 e 2016. Resultados: 20 artigos compuseram o produto final da seleção, sendo 14 artigos de revisão, dois estudos transversais, três estudos de corte e um estudo caso-controle. De modo geral, os padrões alimentares associados à prevenção da Doença de Alzheimer são a Dieta Mediterrânea, Dietary Approaches to Stop Hypertension, Dieta Mediterranean/Dietary Approaches to Stop Hypertension. Também há evidências acerca do retardo da progressão do Alzheimer através de vitaminas antioxidantes. Discussão: entre os fatores alimentares de risco para o desenvolvimento da patologia, estão o elevado consumo de ácidos graxos saturados e trans, carnes vermelhas em excesso e álcool. Conclusão: ficou evidente o papel protetor contra o desenvolvimento da DA através de uma dieta saudável. Alguns nutrientes parecem ser promissores na prevenção e redução da progressão da DA, como as vitaminas C e E e selênio, vitaminas B9 e B12, especialmente quando oriundos de fontes dietéticas e parte de um padrão alimentar saudável.

Objective: To verify if nourishment and nutrition can contribute to the prevention or retardation of the progression of Alzheimer's disease. Methods: An integrative review was carried out in the databases: Lilacs, Medline, Capes Periodicals and National Center of Biotechnology Information between the period 2006 and 2016. Results: 20 articles composed the final product of the selection, 14 articles of 2 cross-sectional studies, 3 cohort studies and 1 case-control study. In general, the dietary patterns associated with Alzheimer's disease prevention are the Mediterranean diet, Dietary Approaches to Stop Hypertension, Mediterranean Diet/Approach to Stop Hypertension. There is also evidence of delayed Alzheimer's proration through antioxidant vitamins. Discussion: Among the dietary risk factors for the development of the pathology are the high consumption of saturated and trans fatty acids, excess red meat and alcohol. Conclusion: The protective role against the development of AD through a healthy diet became evident. Some nutrients appear to be promising in preventing and reducing the progression of AD, such as vitamins C and E and selenium, vitamins B9 and B12, especially when derived from dietary sources and part of a healthy eating pattern.

Impact of oral gut decontamination on Staphylococcus aureus colonisation in patients undergoing allogeneic haematopoietic stem cell transplantation.

Recipients of allogeneic haematopoietic stem cell transplantation (allo-HSCT) are severely immunocompromised and are at increased risk of infection. In this prospective, observational, single-centre study including 110 allo-HSCT recipients, the rate of Staphylococcus aureus colonisation was reduced from 11.8% to 0% (P <0.001) following peritransplant oral gut decontamination. No invasive S. aureus infections were observed.

Efeitos de um programa de educação nutricional domiciliar para adolescentes asmáticos / Effects of a domiciliary nutritional education program for asthmatic adolescents

Objetivo: investigar os efeitos de um programa de educação alimentar e nutricional sobre os sintomas daasma em escolares asmáticos. Método: estudo descritivo, de intervenção, ensaio clínico. Foram realizadasatividades de educação nutricional domiciliar, ao longo de seis meses, com oito escolares asmáticos. Avaliouse,no momento de pré-intervenção e no pós-intervenção, para fins de comparação. Resultados: houveinadequação na ingestão alimentar dos escolares, com incremento no consumo de frutas, legumes, verduras eleguminosas durante a pesquisa. A avaliação da asma, como bem controlada, passou de 25% para 62,5%, e opercentual de escolares com mais de 12 crises de asma ao mês diminuiu de 37,5% para 12,5%. Conclusão:pode-se inferir que houve efetiva melhora do comportamento alimentar e aumento no controle da asma comdiminuição da frequência e intensidade da asma.

The roles of the nitrate reductase NarGHJI, the nitrite reductase NirBD and the response regulator GlnR in nitrate assimilation of Mycobacterium tuberculosis.

Mycobacterium tuberculosis can utilize various nutrients including nitrate as a source of nitrogen. Assimilation of nitrate requires the reduction of nitrate via nitrite to ammonium, which is then incorporated into metabolic pathways. This study was undertaken to define the molecular mechanism of nitrate assimilation in M. tuberculosis. Homologues to a narGHJI-encoded nitrate reductase and a nirBD-encoded nitrite reductase have been found on the chromosome of M. tuberculosis. Previous studies have implied a role for NarGHJI in nitrate respiration rather than nitrate assimilation. Here, we show that a narG mutant of M. tuberculosis failed to grow on nitrate. A nirB mutant of M. tuberculosis failed to grow on both nitrate and nitrite. Mutant strains of Mycobacterium smegmatis mc(2)155 that are unable to grow on nitrate were isolated. The mutants were rescued by screening a cosmid library from M. tuberculosis, and a gene with homology to the response regulator gene glnR of Streptomyces coelicolor was identified. A DeltaglnR mutant of M. tuberculosis was generated, which also failed to grow on nitrate, but regained its ability to utilize nitrate when nirBD was expressed from a plasmid, suggesting a role of GlnR in regulating nirBD expression. A specific binding site for GlnR within the nirB promoter was identified and confirmed by electrophoretic mobility shift assay using purified recombinant GlnR. Semiquantitative reverse transcription PCR, as well as microarray analysis, demonstrated upregulation of nirBD expression in response to GlnR under nitrogen-limiting conditions. In summary, we conclude that NarGHJI and NirBD of M. tuberculosis mediate the assimilatory reduction of nitrate and nitrite, respectively, and that GlnR acts as a transcriptional activator of nirBD.

Human cardiac phospholipase D activity is tightly controlled by phosphatidylinositol 4,5-bisphosphate.

Phospholipase D (PLD) plays a central role in receptor-mediated breakdown of choline phospholipids and formation of phosphatidic acid (PA), an important regulator of cardiac function. However, specific mechanisms that regulate myocardial PLD activity remain largely unknown, particularly in the human heart. We hypothesized that phosphatidylinositol 4,5-bisphosphate (PIP2), best known as substrate for phospholipase C (PLC) isozymes, plays a critical role in regulating myocardial PLD activity. We examined the effect of PIP2 on human myocardial PLD activity in vitro by utilizing a fluorescence HPLC assay. PIP2 increased 10-fold the maximal activity of a partially solubilized PLD from human atrial myocardium. PIP2-stimulated PLD activity was accompanied by a consecutive increase in diacylglycerol, indicating dephosphorylation of PA by PA phosphohydrolase. Likewise, phosphatidylinositol 3,4,5-trisphosphate, which is produced from PIP2 by phosphatidylinositol 3-kinase, increased PLD activity with about the same potency but with somewhat lower efficacy. In contrast, other phospholipids were ineffective, indicating that the action of PIP2 on PLD is highly specific. Neomycin, a high-affinity ligand of PIP2, inhibited PLD activity in human atrial myocardium, but had no effect on the activity of partially solubilized enzyme. The addition of PIP2 restored the sensitivity of solubilized PLD to neomycin inhibition, indicating that neomycin inhibits PLD activity by binding to endogenous PIP2. Our results demonstrate a critical role for PIP2 in human cardiac PLD activity and suggest that PIP2 synthesis (by phosphatidylinositol 4-phosphate 5-kinase) and hydrolysis (by PIP2-specific PLC) could be important determinants in regulating PLD signal transduction in the human heart.