Produção de clamidósporos de Pochonia chlamydosporia em diferentes substratos

Clamidósporos são estruturas de sobrevivência do fungo nematófago Pochonia chlamydosporia. Objetivou-se, com este trabalho, avaliar diferentes substratos, teores de água e formas de inóculo para a produção in vitro de clamidósporos de P. chlamydosporia. Inicialmente, testaram-se os substratos grãos de milho triturado, grãos de arroz e casca de café e os tipos de inóculo meio líquido concentrado ou diluído (1:40) e discos de cultura, colonizados por P. chlamydosporia. Posteriormente, testou-se o substrato grãos de milho triturado suplementado ou não com caldo de batata com os inóculos do fungo nas formas de discos de micélio ou suspensão aquosa. As maiores produções de clamidósporos g-1 de substrato foram obtidas nos substratos grãos de milho triturado e casca de café, e as melhores formas de inóculo foram meio líquido diluído (1:40) e discos de micélio. A suplementação do substrato grãos de milho triturado com caldo de batata não aumentou a produção de clamidósporos do fungo e a melhor forma de inóculo do fungo foi a de discos de micélio. Além disso, testaram-se os substratos canjica, grãos de arroz, casca de café e fibra de coco, umedecidos com diferentes quantidades de água; o substrato casca de café umedecido por diferentes períodos; a casca de café enriquecida ou não com sacarose e farinha de arroz e o substrato grãos de arroz esterilizado no forno micro-ondas com diferentes quantidades de água. Apenas o substrato grãos de arroz, em todas as quantidades de água testadas, apresentou maior produção de clamidósporos. Não houve diferença na produção de clamidósporos no substrato casca de café umedecido por diferentes períodos e apenas quando a casca de café foi enriquecida com farinha de arroz, apresentou maior média de clamidósporos g-1 de substrato. Todos os tratamentos apresentaram grande produção de clamidósporos g-1 quando o substrato utilizado foi o de grãos de arroz tratados no forno de micro-ondas. O melhor meio de cultivo de P. chlamydosporia para a produção de clamidósporos foi o substrato contendo grãos de arroz.Chlamydospores are survival structures of the nematophagous fungus Pochonia chlamydosporia. The objective of this study was to evaluate different substrates, different contents of water and kinds of inoculum for the production of Pochonia chlamydosporia chlamydospores. The substrates evaluated were: milled maize, rice grains and coffee husk; all of them were inoculated with disks of culture, concentrated liquid media or diluted liquid media (1:40) colonized by P. chlamydosporia. Besides, the substrates milled maize, supplemented or not with potato broth, and inoculated with fungus disks or aqueous suspension were evaluated. Milled maize and coffee husk were the best substrates for chlamydospores production. The best inoculum kinds were disks of culture and diluted liquid media (1:40) colonized by P. chlamydosporia. The supplementation of milled maize with potato broth did not improve the production of chlamydospores, and the best inoculum form was disks of fungus. Moreover, it was also studied the substrates rice grains, coffee husk and coconut fibers with different rates of water; the substrate coffee husk moistened by different periods; the supplementation of coffee husk with rice flour or dextrose; and the substrate grains of rice sterilized in microwave oven with different rates of water. The substrate grains of rice, in all of the amounts of water tested showed increase in production of chlamydospores. There was no difference in the chlamydospores production of the coffee husk substrate, when moistened by different periods of time and only when it was supplemented with rice flour it showed higher mean of clamydospores per gram of substrate. All treatments evaluated showed higher production of chlamydospores in the substrate grains of rice treated in the microwave oven. The best substrate for the production of chlamydospores of P. chlamydosporia was the rice grains

Absent B cells, agammaglobulinemia, and hypertrophic cardiomyopathy in folliculin-interacting protein 1 deficiency

Agammaglobulinemia is the most profound primary antibody deficiency that can occur due to an early termination of B-cell development. We here investigated 3 novel patients, including the first known adult, from unrelated families with agammaglobulinemia, recurrent infections, and hypertrophic cardiomyopathy (HCM). Two of them also presented with intermittent or severe chronic neutropenia. We identified homozygous or compound-heterozygous variants in the gene for folliculin interacting protein 1 (FNIP1), leading to loss of the FNIP1 protein. B-cell metabolism, including mitochondria! numbers and activity and phosphatidylinositol 3-kinase/AKT pathway, was impaired. These defects recapitulated the Fnip1(-/-) animal model. Moreover, we identified either uniparental disomy or copy-number variants (CNVs) in 2 patients, expanding the variant spectrum of this novel inborn error of I immunity. The results indicate that FNIP1 deficiency can be caused by complex genetic mechanisms and support the clinical utility of exome sequencing and CNV analysis in patients with broad phenotypes, including agammaglobulinemia and HCM. FNIP1 deficiency is a novel inborn error of immunity characterized by early and severe B-cell development defect, agammaglobulinemia, variable neutropenia, and HCM. Our findings elucidate a functional and relevant role of FNIP1 in B-cell development and metabolism and potentially neutrophil activity.Molecular Technology and Informatics for Personalised Medicine and Healt

Primary resistance to integrase strand-transfer inhibitors in Europe

Objectives: The objective of this study was to define the natural genotypic variation of the HIV-1 integrase gene across Europe for epidemiological surveillance of integrase strand-transfer inhibitor (InSTI) resistance. Methods: This was a multicentre, cross-sectional study within the European SPREAD HIV resistance surveillance programme. A representative set of 300 samples was selected from 1950 naive HIV-positive subjects newly diagnosed in 2006-07. The prevalence of InSTI resistance was evaluated using quality-controlled baseline population sequencing of integrase. Signature raltegravir, elvitegravir and dolutegravir resistance mutations were defined according to the IAS-USA 2014 list. In addition, all integrase substitutions relative to HXB2 were identified, including those with a Stanford HIVdb score=10 to at least one InSTI. To rule out circulation of minority InSTIresistant HIV, 65 samples were selected for 454 integrase sequencing. Results: For the population sequencing analysis, 278 samples were retrieved and successfully analysed. No signature resistance mutations to any of the InSTIswere detected. Eleven (4%) subjects hadmutations at resistance-associated positions with an HIVdb score =10. Of the 56 samples successfully analysed with 454 sequencing, no InSTI signature mutationsweredetected, whereas integrase substitutionswithanHIVdbscore=10were found in8(14.3%) individuals. Conclusions:No signature InSTI-resistant variantswere circulating in Europe before the introduction of InSTIs. However, polymorphisms contributing to InSTI resistancewere not rare. As InSTI use becomes more widespread, continuous surveillance of primary InSTI resistance is warranted. These data will be key to modelling the kinetics of InSTI resistance transmission in Europe in the coming years. © The Author 2015. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved

Safety of hospital discharge before return of bowel function after elective colorectal surgery

© 2020 BJS Society Ltd Published by John Wiley & Sons LtdBackground: Ileus is common after colorectal surgery and is associated with an increased risk of postoperative complications. Identifying features of normal bowel recovery and the appropriateness for hospital discharge is challenging. This study explored the safety of hospital discharge before the return of bowel function. Methods: A prospective, multicentre cohort study was undertaken across an international collaborative network. Adult patients undergoing elective colorectal resection between January and April 2018 were included. The main outcome of interest was readmission to hospital within 30 days of surgery. The impact of discharge timing according to the return of bowel function was explored using multivariable regression analysis. Other outcomes were postoperative complications within 30 days of surgery, measured using the Clavien–Dindo classification system. Results: A total of 3288 patients were included in the analysis, of whom 301 (9·2 per cent) were discharged before the return of bowel function. The median duration of hospital stay for patients discharged before and after return of bowel function was 5 (i.q.r. 4–7) and 7 (6–8) days respectively (P < 0·001). There were no significant differences in rates of readmission between these groups (6·6 versus 8·0 per cent; P = 0·499), and this remained the case after multivariable adjustment for baseline differences (odds ratio 0·90, 95 per cent c.i. 0·55 to 1·46; P = 0·659). Rates of postoperative complications were also similar in those discharged before versus after return of bowel function (minor: 34·7 versus 39·5 per cent; major 3·3 versus 3·4 per cent; P = 0·110). Conclusion: Discharge before return of bowel function after elective colorectal surgery appears to be safe in appropriately selected patients