Multiplex PCR identification of eight clinically relevant Candida species

Invasive fungal infections, specifically candidemia, constitute major public health problems with high mortality rates. Therefore, in the last few years, the development of novel diagnostic methods has been considered a critical issue. Herein we describe a multiplex PCR strategy allowing the identification of 8 clinically relevant yeasts of the Candida genus, namely C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, C. krusei, C. guilliermondii, C. lusitaniae and C. dubliniensis. This method is based on the amplification of two fragments from the ITS1 and ITS2 regions by the combination of 2 yeast-specific and 8 species-specific primers in a single PCR reaction. Results from the identification of 231 clinical isolates are presented pointing to the high specificity of this procedure. Furthermore, several Candida isolates were identified directly from clinical specimens which also attests to the method's direct laboratory application. The results from the multiplex reactions with other microorganisms that usually co-infect patients also confirmed its high specificity in the identification of Candida species. Moreover, this method is simple and presents a sensitivity of approximately 2 cells per ml within 5 hours. Furthermore, it allows discrimination of individual Candida species within polyfungal samples. This novel method may therefore provide a clinical diagnostic procedure with direct applicability.Agostinho Carvalho was financially supported by a fellowship from Fundação para a Ciência e Tecnologia, Portugal (contract SFRH/BD/11837/2003). This study was supported by Fundação para a Ciência e Tecnologia, Portugal (POCI/SAU-ESP/61080/2004)

Functional genetic variants in ATG10 are associated with acute myeloid leukemia

Acute myeloid leukemia (AML) is the most common acute leukemia, characterized by a heterogeneous genetic landscape contributing, among others, to the occurrence of metabolic reprogramming. Autophagy, a key player on metabolism, plays an essential role in AML. Here, we examined the association of three potentially functional genetic polymorphisms in the ATG10 gene, central for the autophagosome formation. We screened a multicenter cohort involving 309 AML patients and 356 healthy subjects for three ATG10 SNPs: rs1864182T>G, rs1864183C>T and rs3734114T>C. The functional consequences of the ATG10 SNPs in its canonical function were investigated in vitro using peripheral blood mononuclear cells from a cohort of 46 healthy individuals. Logistic regression analysis adjusted for age and gender revealed that patients carrying the ATG10rs1864182G allele showed a significantly decreased risk of developing AML (OR [odds ratio] = 0.58, p = 0.001), whereas patients carrying the homozygous ATG10rs3734114C allele had a significantly increased risk of developing AML (OR = 2.70, p = 0.004). Functional analysis showed that individuals carrying the ATG10rs1864182G allele had decreased autophagy when compared to homozygous major allele carriers. Our results uncover the potential of screening for ATG10 genetic variants in AML prevention strategies, in particular for subjects carrying other AML risk factors such as elderly individuals with clonal hematopoiesis of indeterminate potential.This research was funded by FEDER and Foundation for Science and Technology (FCT), grant number POCI-01-0145-FEDER-028159 and POCI-01-0145-FEDER-030782); by Fondo de Investigaciones Sanitarias (Madrid, Spain), grant number ISCIII-FEDER PI20/01845, ISCIII-FEDER PI12/02688, and ISCIII-FEDER PI17/02276. B.S.M. was funded by FCT, grant number DL 57/2016. A.C.A. was funded by FCT, grant number POCI-01-0145-FEDER-028159. C.C. was funded by FCT, grant number CEECIND/04058/2018. A.C. was funded by FCT, grant number CEECIND/03628/2017

Development, economic viability and attributes of lamb carcass from confined animals fed on different amounts of crude glycerin

<p>The current study aims to assess the effect from crude glycerin inclusion (0, 7, 14, and 21% dry matter) in the diet of slaughtered lamb on their development, nutrient consumption, biometrical measures, diet economic viability and carcass features. Thirty two (32) non-castrated male Texel lambs were used in the study, they presented mean initial weight 15.9 ± 4.1 kilos and were distributed in casual outlining. They were fed with four treatments, with 8 repetitions. Animals were slaughtered when they reached approximately 35 kilos. The mean total weight gain was 20.72 kilos and mean daily weight gain was 260 grams. No changes resulted from glycerin use. The carcass performance was similar among treatments (P&gt;0.05) and the cold carcass performance (CCP) was 44.68%. There were no effects (P&gt;0.05) on the loin eye area (LEA) and on fat thickness (FT); they showed averages of 13.66 cm2 and 0.84 mm, respectively. Nutrition cost per animal during the whole confinement period varied between R$82.60 (eighty-two Reais and forty-eight cents) to R$92.48. The smallest nutrition amount consisted of 21% crude glycerin. The gross profit ranged from R$30.75 to R$ 34.01 per animal, for feed without glycerin and 21% glycerin, respectively. Animal development was not impacted by glycerin introduction, even with decrease on dry and organic mass consumption. The result showed that crude glycerin inclusion might be used in lambs’ diet. Whenever there are big amounts of feed involved in the process, the 21% crude glycerin addition may be an interesting cost reduction. Seventy eight percent (78%) glycerol crude glycerin to replace corn-based feed in confined lambs’ diet appeared to be nutritionally and economically viable.</p><p><strong> </strong></p

O EFEITO DA SUPLEMENTAÇÃO COM ZINCO ORGÂNICO NO GANHO DE PESO DE BORREGAS TEXEL

A demanda por proteína animal e a facilidade de criação tornam a ovinocultura uma atividade em evidência, e melhorar o desempenho dos animais uma necessidade cada vez maior. O objetivo deste trabalho foi avaliar o efeito da suplementação com zinco orgânico no desempenho de borregas Texel. Foram selecionadas, aleatoriamente, 46 borregas da raça Texel, com oito meses de idade, pesando em média 41 kg PV. Os animais foram divididos em dois grupos experimentais (23 animais/ grupo): grupo tratado (GZn) suplementado com mistura mineral e adição de Zn orgânico e grupo controle (Gc) suplementado somente com a mistura mineral. Os grupos ficaram em 2 piquetes de 4 ha de Brachiaria brizantha, separados, e foram alternados mensalmente, o Gc recebeu 500g da mistura mineral no aprisco e o GZn além de receber 500g da mistura mineral foi suplementado com 0,5g/ animal/ dia de Zn ogânico por via oral durante o período experimental de 180 dias. O consumo da mistura mineral foi de 22 g/ animal/ dia, nos dois grupos, estando dentro do consumo esperado de 10 a 28 g/ animal/ dia. O ganho de peso no período avaliado foi superior nos animais suplementados com Zn orgânico, 38,3 g/dia no GZn e 8,6 g/dia no Gc, sendo a diferença estatisticamente significativa (p≤ 0,05). O efeito da suplementação com zinco orgânico no desempenho de borregas Texel foi positivo, com aumento do ganho de peso médio diário nos animais suplementados

Chemical composition and losses of nutrients involved in sugarcane ensiling with microbial and chemical additives

There is increasing interest in sugarcane silage as an alternative supplemental feed for cattle, but high losses during the fermentation can affect the viability of this option. The present study evaluated the effects of additives on the composition and nutrient losses in sugarcane ensiling by applying the following six treatments: without additive (CON), with 2 × 106 cfu/g of Lactobacillus buchneri alone (LB) or associated with 2 × 106 cfu/g of Lactobacillus plantarum (LB/PLA), 1.0% urea (LB/URE), 0.5% virgin lime (LB/CAL), and only 0.5% virgin lime (CAL) on an as fed basis. Silos with 18 L capacity were used and the fermentation lasted 104 days. After opening the silos, the dry matter (DM) content of the silage was found to have decreased by 3.9 percentage units on average compared to the initial value. Crude protein (CP) content was higher and neutral detergent fiber (NDF) lower in the (LB/URE) treatment. In the treatments with virgin lime the pH value was higher and the recovery of MS was lower, but equal to that of the treatment LB, in which the effluent losses were greater. The gaseous losses were lower in treatments with only microbial additives. It was concluded that none of the additives evaluated reduce dry matter losses, but the microbial additives reduce gas losses and the virgin lime reduces effluent losses

The C Allele of rs5743836 Polymorphism in the Human TLR9 Promoter Links IL-6 and TLR9 Up-Regulation and Confers Increased B-Cell Proliferation

In humans, allelic variants in Toll-like receptors (TLRs) associate with several pathologies. However, the underlying cellular and molecular mechanisms of this association remain largely unknown. Analysis of the human TLR9 promoter revealed that the C allele of the rs5743836 polymorphism generates several regulatory sites, including an IL-6-responding element. Here, we show that, in mononuclear cells carrying the TC genotype of rs5743836, IL-6 up-regulates TLR9 expression, leading to exacerbated cellular responses to CpG, including IL-6 production and B-cell proliferation. Our study uncovers a role for the rs5743836 polymorphism in B-cell biology with implications on TLR9-mediated diseases and on the therapeutic usage of TLR9 agonists/antagonists

OVINOCULTURA LEITEIRA NO BRASIL: ASPECTOS E FATORES RELACIONADOS À COMPOSIÇÃO, AO CONSUMO E À LEGISLAÇÃO

Ao longo dos últimos anos a ovinocultura leiteira vem ganhando espaço no mercado internacional e nacional brasileiro. Sua exploração mostra-se bastante vantajosa e rentável, visto que, o leite de ovelha carrega consigo características tecnológicas importantes para a indústria de alimentos. Seu elevado potencial de ser transformado em derivados lácteos, que agregam alta qualidade nutricional, atrai investidores e consumidores. As altas concentrações de sólidos totais presentes no leite de ovelha permitem a formulação de queijos, iogurtes e manteigas, de rentabilidade superior a de outras espécies. Fatores como a raça, a alimentação, a estação do ano, o estágio de lactação e o estado sanitário são variáveis que influenciam diretamente na composição do leite de ovelha. No Brasil, ainda não existe legislação específica para controle da qualidade do leite de ovinos, o que demonstra a necessidade de sua criação. São poucos os estudos brasileiros que caracterizam a composição e qualidade do leite de ovelha produzido no Brasil, sendo assim, importante o estudo das suas características e fatores que determinam sua composição. Este trabalho tem como objetivo projetar um delineamento da ovinocultura leiteira brasileira, com enfoque nos aspectos da legislação, da composição e da qualidade do leite

Search for dark matter produced in association with bottom or top quarks in √s = 13 TeV pp collisions with the ATLAS detector

A search for weakly interacting massive particle dark matter produced in association with bottom or top quarks is presented. Final states containing third-generation quarks and miss- ing transverse momentum are considered. The analysis uses 36.1 fb−1 of proton–proton collision data recorded by the ATLAS experiment at √s = 13 TeV in 2015 and 2016. No significant excess of events above the estimated backgrounds is observed. The results are in- terpreted in the framework of simplified models of spin-0 dark-matter mediators. For colour- neutral spin-0 mediators produced in association with top quarks and decaying into a pair of dark-matter particles, mediator masses below 50 GeV are excluded assuming a dark-matter candidate mass of 1 GeV and unitary couplings. For scalar and pseudoscalar mediators produced in association with bottom quarks, the search sets limits on the production cross- section of 300 times the predicted rate for mediators with masses between 10 and 50 GeV and assuming a dark-matter mass of 1 GeV and unitary coupling. Constraints on colour- charged scalar simplified models are also presented. Assuming a dark-matter particle mass of 35 GeV, mediator particles with mass below 1.1 TeV are excluded for couplings yielding a dark-matter relic density consistent with measurements