Current methods to analyze lysosome morphology, positioning, motility and function

Since the discovery of lysosomes more than 70 years ago, much has been learned about the functions of these organelles. Lysosomes were regarded as exclusively degradative organelles, but more recent research has shown that they play essential roles in several other cellular functions, such as nutrient sensing, intracellular signalling and metabolism. Methodological advances played a key part in generating our current knowledge about the biology of this multifaceted organelle. In this review, we cover current methods used to analyze lysosome morphology, positioning, motility and function. We highlight the principles behind these methods, the methodological strategies and their advantages and limitations. To extract accurate information and avoid misinterpretations, we discuss the best strategies to identify lysosomes and assess their characteristics and functions. With this review, we aim to stimulate an increase in the quantity and quality of research on lysosomes and further ground-breaking discoveries on an organelle that continues to surprise and excite cell biologists.Medical Biochemistr

Desempenho de ovinos mantidos com dietas com capim-elefante ensilado com diferentes aditivos Performance of sheep fed diets with elephant grass ensiled with different addictives

Avaliaram-se os efeitos da adição de 15% de casca de café, farelo de cacau ou farelo de mandioca, com base na matéria natural (peso/peso), à forragem verde de capim-elefante no momento da ensilagem sobre o consumo de nutrientes e o desempenho de 20 ovinos Santa Inês em confinamento. Os animais tinham 5 meses de idade, peso inicial de 22,27 ± 3,24 kg e o peso final foi de 28,6 ± 3,78 kg. Utilizou-se um delineamento experimental inteiramente ao acaso, com quatro tratamentos e cinco repetições. Os tratamentos foram: capim-elefante ensilado sem aditivo ou ensilado com 15% de casca de café, farelo de cacau ou farelo de mandioca. A utilização da casca de café, do farelo de cacau e do farelo de mandioca na ensilagem de capim-elefante aumentou o consumo de matéria seca. Os maiores consumos de proteína bruta foram observados nas silagens contendo farelo de cacau e farelo de mandioca. Os consumos de fibra em detergente neutro foram semelhantes e o consumo de nutrientes digestíveis totais foi maior na silagem contendo farelo de mandioca. A conversão alimentar dos animais não foi afetada pela inclusão dos aditivos na ensilagem. A inclusão de farelo de mandioca na ensilagem de capim-elefante aumenta o ganho de peso (140,5 g/dia) em ovinos Santa Inês.<br>The effects of the addition of 15% of different addictives (coffee hulls, cocoa meal and cassava meal) to green elephant grass forage, in natural matter basis (weight/weight), at the ensilage moment on the consumption of nutrients and the performance of twenty Santa Inês sheep under feedlot system were evaluated. The animals had 5.0 months of age, initial weight of 22.27 ± 3.24 kg and final weight of 28.6 ± 3.78 kg. The treatments were: T1 = elephant grass ensiled without addictive; T2 = elephant grass ensiled with 15% coffee hulls; T3 = elephant grass ensiled with 15% cocoa meal; T4 = elephant grass ensiled with 15% cassava meal. The use of the addictives coffee hulls, cocoa meal and cassava meal in elephant grass ensilage increased the dry matter intake. The greatest crude protein intakes were observed with silage containing cocoa meal and cassava meal. Neutral detergent fiber intakes were similar, and the total digestible nutrients intake was higher in silage containing cassava bran. The feed conversion rate of the animals was not affected by the inclusion of addictives in ensilage. Diet containing elephant grass ensiled with cassava meal showed higher weight gain along the period and per day, however, no difference was observed in the feed conversion rate. The addition of cassava meal in ensilage of elephant grass provided weight gain of 140.5 g/day in Santa Inês sheep

Subprodutos industriais na ensilagem de capim-elefante para cabras leiteiras: consumo, digestibilidade de nutrientes e produção de leite Industrial by-products in elephant grass silage for dairy goats: intake, nutrient digestibility and milk yield

O experimento foi conduzido para avaliar o consumo, a digestibilidade aparente e a produção do leite em cabras alimentadas com silagens de capim-elefante contendo subprodutos industriais. Utilizaram-se oito cabras da raça Saanen, distribuídas em dois quadrados latinos 4 &#215; 4, com 4 períodos de 14 dias (10 dias de adaptação e 4 dias para coleta de dados). Os tratamentos foram: silagem de capim-elefante + concentrado; silagem de capim-elefante com 15% de farelo de mandioca + concentrado; silagem de capim-elefante com 15% de casca de café + concentrado; silagem de capim-elefante com 15% farelo de cacau + concentrado. Os maiores consumos de matéria seca (MS), matéria orgânica (MO), carboidratos totais (CT), carboidratos não-fibrosos (CNF) e nutrientes digestíveis totais foram observados entre os animais alimentados com a silagem contendo farelo de mandioca. As silagens com farelo de mandioca ou casca de café promoveram maior consumo de extrato etéreo. Os maiores valores de digestibilidade da MS, MO, EE e CT foram obtidos na dieta contendo silagem de capim-elefante com farelo de mandioca. O valor de digestibilidade dos CNF foi semelhante entre as dietas contendo silagem de capim-elefante com farelo de mandioca, casca de café e farelo de cacau. A digestibilidade de fibra em detergente neutro e de fibra em detergente ácido foi maior para a silagem sem aditivo, com farelo de mandioca ou farelo de cacau. Entre os subprodutos avaliados, o farelo de mandioca é o que promove maiores consumos, digestibilidades dos nutrientes e produção de leite em cabras alimentadas com silagem de capim-elefante.<br>The experiment was carried out to evaluate the intake, apparent digestibility and the milk yield of goats fed elephant grass silages containing industrial by-products. Eight Saanen breed goats were used, distributed in two 4 &#215; 4 Latin squares, with four 14-day periods (ten days of adaptation and four days for data collection). The treatments were: elephant grass silage + concentrate; elephant grass silage containing 15% cassava meal + concentrate; elephant grass silage containing 15% coffee hulls + concentrate; elephant grass silage containing 15% cocoa meal + concentrate. The greatest intakes of dry matter (DM), organic matter (OM), total carbohydrates (TC), nonfiber carbohydrate (NFC) and digestible total nutrients were observed in the animals fed silage containing cassava meal. The greatest intake of ether extract was verified in the silages with cassava meal or coffee hulls. The greatest digestibility values of the DM, OM, EE and TC were obtained in the diet containing elephant grass silage with cassava meal. The NFC digestibility value was similar among diets containing elephant grass silage with cassava meal, coffee hulls and cocoa meal. The neutral detergent fiber and acid detergent fiber digestibility was greater in the silage without additive, with cassava meal or cocoa meal. Greater intake, digestibility coefficients and milk yield were obtained in goats fed elephant grass silage containing cassava meal

Treatment of Multisystem Inflammatory Syndrome in Children

BACKGROUND Evidence is urgently needed to support treatment decisions for children with multisystem inflammatory syndrome (MIS-C) associated with severe acute respiratory syndrome coronavirus 2. METHODS We performed an international observational cohort study of clinical and outcome data regarding suspected MIS-C that had been uploaded by physicians onto a Web-based database. We used inverse-probability weighting and generalized linear models to evaluate intravenous immune globulin (IVIG) as a reference, as compared with IVIG plus glucocorticoids and glucocorticoids alone. There were two primary outcomes: the first was a composite of inotropic support or mechanical ventilation by day 2 or later or death; the second was a reduction in disease severity on an ordinal scale by day 2. Secondary outcomes included treatment escalation and the time until a reduction in organ failure and inflammation. RESULTS Data were available regarding the course of treatment for 614 children from 32 countries from June 2020 through February 2021; 490 met the World Health Organization criteria for MIS-C. Of the 614 children with suspected MIS-C, 246 received primary treatment with IVIG alone, 208 with IVIG plus glucocorticoids, and 99 with glucocorticoids alone; 22 children received other treatment combinations, including biologic agents, and 39 received no immunomodulatory therapy. Receipt of inotropic or ventilatory support or death occurred in 56 patients who received IVIG plus glucocorticoids (adjusted odds ratio for the comparison with IVIG alone, 0.77; 95% confidence interval [CI], 0.33 to 1.82) and in 17 patients who received glucocorticoids alone (adjusted odds ratio, 0.54; 95% CI, 0.22 to 1.33). The adjusted odds ratios for a reduction in disease severity were similar in the two groups, as compared with IVIG alone (0.90 for IVIG plus glucocorticoids and 0.93 for glucocorticoids alone). The time until a reduction in disease severity was similar in the three groups. CONCLUSIONS We found no evidence that recovery from MIS-C differed after primary treatment with IVIG alone, IVIG plus glucocorticoids, or glucocorticoids alone, although significant differences may emerge as more data accrue. Copyright © 2021 Massachusetts Medical Society