Molecular diagnosis of Eimeria species affecting naturally infected Gallus gallus.

Abstract - We used PCR to test various protocols and define a technique for DNA extraction directly from chicken-shed stool samples for the identification of Eimeria species that parasitize birds. It was possible to extract and amplify DNA of seven Eimeria species from field stool samples, using both protocols tested; extractions made with phenol/chloroform protocols gave the best results. The primers were specific and sensitive, allowing amplification of samples containing as few as 20 oocysts, both in individual and in a multiplex PCR. Individualized PCR with the phenol/chloroform DNA extraction protocol detected a larger number of Eimeria species. Molecular diagnosis was found to be practical and precise, and can be used for monitoring and epidemiological studies of Eimeria

Adipose Tissue-Derived Stem Cells and the Importance of Animal Model Standardization for Pre-Clinical Trials

ABSTRACTStem cells are undifferentiated cells and can self-renew and differentiate into various cell types, besides having immuno-modulating properties and paracrine effects in response to tissue injury, and may therefore treat injuries and diseases or even replace damaged or lost cells. Adipose tissue is an attractive source of adult stem cells, since the human body has a large reserve that is obtained in large amounts by minimally invasive methods. Interest in these cells has been increasing steadily due to their properties and possible applications in regenerative medicine and cell therapy. A large part of these investigations are focused on cardiovascular diseases, which are a leading cause of morbidity and mortality worldwide. Although in recent years treatments have advanced in cardiology, the development of new therapies to recover the damaged tissue still remains one of the main goals of cardiac research. However, to achieve effective results, in vivo and in vitro animal models for preclinical studies and consequently for application in humans must be standardized. The development of preclinical models in large animals requires the use of well-characterized animal cell lines, similar to human cells, and the use of the porcine model represents a great advantage for preclinical translational research

Análise de repetições em dados biológicos

The decoding of the genomes has created new challenges on the scientific community linked to the area of computation and information technologies. Daily, new data is added to numerous databases with billions of records, coming from more advanced equipment, helping in decoding the genomes. Determine how important and relevant are these data in order to find value-added information and obviously turn them into knowledge,is the main challenge for the bioinformatics research community. The analysis of genomes and proteomes of several organisms allow us to observe the behaviour at the evolution of species. In this study, our focus goes to a particular aspect of this analysis: the repetition of some codons and their amino acids inside several orthologous genes in eukaryotic organisms. Belonging to different stages of evolution, the main objective focuses on achieving results on the evolution of these repetitions over millions of years. We now know that these repetitions in humans are the source of several neurodegenerative diseases among others. This analysis will verify the conservation or repression, of these repetitions throughout the process of speciation as well at the level of relationship that may exist between these repetitions and those diseases. For this study we have developed an algorithm for A descodificação dos genomas veio criar novos desafios na comunidade científica ligada à área da computação e da informática. Diariamente são alimentadas inúmeras bases de dados com biliões de registos provenientes de equipamentos cada vez mais evoluídos, que auxiliam na descodificação dos genomas. Determinar o quão importante e relevante são esses dados, de forma a retirar valor acrescentado – informação, e obviamente transformá-los em conhecimento, é o grande desafio actual para a comunidade de investigadores de bioinformática. A análise de genomas, bem como dos proteomas dos vários organismos permitem-nos observar o comportamento ao nível da evolução das espécies. Neste estudo focamos a atenção num aspecto particular dessa análise: as repetições de determinados codões e dos respectivos aminoácidos nos vários organismos eucariotas, especificamente em genes ortólogos. Pertencente a várias fases da evolução das espécies, o objectivo principal centra-se na obtenção de resultados quanto à evolução dessas repetições ao longo de milhões de anos. Sabemos hoje que essas repetições no ser humano são a causa de diversas doenças neuro-degenerativas, entre outras, pelo que esta análise permitirá verificar o estado de conservação ou repressão, dessas repetições ao longo do processo de especiação, bem como ao nível do relacionamento que poderá existir entre essas repetições e as doenças nos seres superiormente evoluídos. Para este estudo foi desenvolvido um algoritmo de detecção de padrões de repetição, que possibilita uma análise detalhada da localização de uma determinada sequência, bem como das sequências que melhor se ajustam ao padrão de repetição inicial.Centro de Estudos em Educação, Tecnologias e Saúd

Endothelial Dysfunction In Cardiovascular And Endocrine-metabolic Diseases: An Update.

The endothelium plays a vital role in maintaining circulatory homeostasis by the release of relaxing and contracting factors. Any change in this balance may result in a process known as endothelial dysfunction that leads to impaired control of vascular tone and contributes to the pathogenesis of some cardiovascular and endocrine/metabolic diseases. Reduced endothelium-derived nitric oxide (NO) bioavailability and increased production of thromboxane A2, prostaglandin H2 and superoxide anion in conductance and resistance arteries are commonly associated with endothelial dysfunction in hypertensive, diabetic and obese animals, resulting in reduced endothelium-dependent vasodilatation and in increased vasoconstrictor responses. In addition, recent studies have demonstrated the role of enhanced overactivation of β-adrenergic receptors inducing vascular cytokine production and endothelial NO synthase (eNOS) uncoupling that seem to be the mechanisms underlying endothelial dysfunction in hypertension, heart failure and in endocrine-metabolic disorders. However, some adaptive mechanisms can occur in the initial stages of hypertension, such as increased NO production by eNOS. The present review focuses on the role of NO bioavailability, eNOS uncoupling, cyclooxygenase-derived products and pro-inflammatory factors on the endothelial dysfunction that occurs in hypertension, sympathetic hyperactivity, diabetes mellitus, and obesity. These are cardiovascular and endocrine-metabolic diseases of high incidence and mortality around the world, especially in developing countries and endothelial dysfunction contributes to triggering, maintenance and worsening of these pathological situations.44920-3

Endothelial dysfunction in cardiovascular and endocrine-metabolic diseases: an update

The endothelium plays a vital role in maintaining circulatory homeostasis by the release of relaxing and contracting factors. Any change in this balance may result in a process known as endothelial dysfunction that leads to impaired control of vascular tone and contributes to the pathogenesis of some cardiovascular and endocrine/metabolic diseases. Reduced endothelium-derived nitric oxide (NO) bioavailability and increased production of thromboxane A2, prostaglandin H2 and superoxide anion in conductance and resistance arteries are commonly associated with endothelial dysfunction in hypertensive, diabetic and obese animals, resulting in reduced endothelium-dependent vasodilatation and in increased vasoconstrictor responses. In addition, recent studies have demonstrated the role of enhanced overactivation of β-adrenergic receptors inducing vascular cytokine production and endothelial NO synthase (eNOS) uncoupling that seem to be the mechanisms underlying endothelial dysfunction in hypertension, heart failure and in endocrine-metabolic disorders. However, some adaptive mechanisms can occur in the initial stages of hypertension, such as increased NO production by eNOS. The present review focuses on the role of NO bioavailability, eNOS uncoupling, cyclooxygenase-derived products and pro-inflammatory factors on the endothelial dysfunction that occurs in hypertension, sympathetic hyperactivity, diabetes mellitus, and obesity. These are cardiovascular and endocrine-metabolic diseases of high incidence and mortality around the world, especially in developing countries and endothelial dysfunction contributes to triggering, maintenance and worsening of these pathological situations449920932CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informaçã

Daily vaginal temperature in Girolando cows from three different genetic composition under natural heat stress.

The present trial evaluated the effect of crossbred composition and Temperature and Humidity Index (THI) on vaginal temperature (VT) of Girolando dairy cows maintained under tropical pasture during warm seasons. The VT was monitored along 41 to 96 h in 615 Girolando cows with different Holstein (H) × Gir genetic composition (1/2 H = 284, 3/4 H = 248, and 7/8 H = 83) from six Brazilian farms in the summer of 2016 and 2017. VT of each cow at each hour of the day and the respective THI were averaged per hour across all monitoring days to generate an averaged value for VT and THI during 24 h. A linear mixed model with repeated measures using restricted maximum likelihood (REML) method for (co)variance components estimation procedure was employed. The final model adjusted the VT for the effects of cow, time, THI, farm, year, pregnancy status, body condition score (BCS), milk yield, genetic composition, and genetic composition*time interaction. Fixed effects were evaluated by ANOVA and tested with Tukey test in R software version 3.6.1 (R Core Team, 2019). Overall mean of VT, air temperature (AT), and THI were 39.06 ± 0.52 °C, 25.63 ± 0.40 °C, and 75.06 ± 3.96, respectively. VT had moderate positive correlation with THI (r² = 0.45, P 0.05). The daily average VT was affected by genetic composition (P < 0.001) with highest temperature for 3/4 H (39.08 ± 0.06 °C a) and 7/8 H (39.09 ± 0.06 °C a) and lowest temperature for 1/2 H (38.95 ± 0.06 °C b). The difference of VT among the three crossbred groups varied in function of the time of the day, from 12:00 to 20:00 h (P < 0.001), with 3/4 Holstein and 7/8 Holstein cows reaching similar VT, above to the upper limit 39.1 °C and higher than 1/2 Holstein cows during all this period. In conclusion, Girolando cows are sensitive to heat stress in tropical condition during warm seasons. Moreover, Girolando cows with genetic composition higher than 3/4 Holstein display reduced thermoregulatory efficiency. Therefore, Girolando cows in tropical dairy farms require strategies to mitigate heat stress according to their genetic composition

Polymorphisms in the leptin gene in divergent swine breeds

Investigou-se a exist?ncia de polimorfismo no gene da leptina (gene da obesidade) entre varr?es da ra?a nativa Piau (porco tipo banha) e matrizes mesti?as de ra?as comerciais (Landrace/Large White e Landrace/Large White com Pietrain), selecionadas para peso e precocidade. Oito pares de primers foram desenhados a partir da seq??ncia dispon?vel no GenBank (U66254), usada, neste trabalho, como seq??ncia de refer?ncia. Amostras de DNA foram extra?das de c?lulas sang??neas brancas utilizando-se solu??o de fenol:clorof?rmio, ap?s tratamento com proteinase K. Os fragmentos gerados por amplifica??o da rea??o em cadeia da polimerase foram purificados e seq?enciados em seq?enciador autom?tico. As seq??ncias de nucleot?deos, obtidas a partir do DNA das ra?as comerciais de su?no, apresentaram maior similaridade com a seq??ncia de refer?ncia, e as seq??ncias geradas a partir do DNA dos animais nativos divergiram de ambas em algumas posi??es. Dos 28 polimorfismos encontrados, oito foram observados em apenas uma das tr?s seq??ncias geradas a partir do DNA das ra?as nativas. Doze estavam presentes em duas seq??ncias, e os oito polimorfismos restantes foram encontrados nos tr?s animais nativos.Funda??o de Amparo ? Pesquisa do Estado de Minas Gerais (FAPEMIG)Empresa de Pesquisa Agropecu?ria de Minas Gerais (EPAMIG)Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq)Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES)Leptin gene (obese gene) polymorphism was investigated in Piau boars (a fat, native breed) and sows from commercial strains (Landrace/Large White and Landrace/Large White by Pietrain) chosen for rapid growth and early sexual maturity. Eight pairs of primers designed using the sequence available from GenBank (access n? U66254) were identified as the reference sequence in this project. DNA samples were extracted from white blood cells using phenol:chloroform solution, after treatment with proteinase K. Fragments generated by amplification of the Polymerase Chain Reaction were purified and sequenced in an automatic sequencer. Nucleotide sequences obtained from DNA of commercial swine breeds were similar to the reference sequence; whereas sequences generated from native breed DNA diverged from the reference sequence and from domestic breed DNA. Of the 28 polymorphisms found, eight were observed in only one of the three sequences generated from DNA of native breeds. Twelve polymorphisms were present in two sequences and the eight remaining polymorphisms were found in all three categories of DNA

Identification of mesenchymal progenitor niches from liver of embryo and fetuses of canines: a source of stem cells for cell therapy

As células-tronco (CT) derivadas dos tecidos fetais (TF) foram as mais recentes descobertas entre as CT, e ultimamente tem demonstrado amplo potencial terapêutico, dentre os TF o fígado fetal (FF) apresenta grande potencial terapêutico. Este órgão durante o período fetal em mamíferos é um nicho hematopoético transitório, sendo o principal órgão responsável pela hematopoese no feto, além de contribuir com a formação do nicho definitivo na medula óssea adulta, portanto pode ser considerado um nicho de células-tronco mesenquimais (CTM) e progenitores. No entanto, pouco se sabe sobre a localização destas células no FF, desta forma o presente estudo visa identificar o nicho de CTM e progenitores em FF de cães, a fim de contribuir com as técnicas de isolamento e extração celular. Em conjunto foi realizada a verificação da expressão do fator de transcrição Oct-3/4 e da proteína delta polimerase do DNA (PCNA). Para a análise foram utilizados cinco embriões e 11 fetos caninos com idades gestacionais variando de 25-60 dias. Os resultados elucidaram a partir de 25 dias de gestação o FF apresentou-se volumoso e composto por todas as estruturas típicas, dentre elas a tríade portal, ductos biliares e ramos das artérias hepáticas. Com 30 dias de gestação foram identificados os primeiros sitos de progenitores mesenquimais (PM) enquanto que aos 60 dias os nichos estavam completamente formados com localização semelhante ao fígado adulto (FA). No entanto, células imunopositivas para Oct-3/4 não foram identificadas; sendo assim, destacamos que o FF é uma fonte de PM, apresentando-se como uma alternativa para a utilização terapêutica, bem como para os estudos da biologia do desenvolvimento das CTM e progenitores

Predicting range shifts of African apes under global change scenarios

Aim: Modelling African great ape distribution has until now focused on current or past conditions, while future scenarios remain scarcely explored. Using an ensemble forecasting approach, we predicted changes in taxon-specific distribution under future scenarios of climate, land use and human populations for (1) areas outside protected areas (PAs) only (assuming complete management effectiveness of PAs), (2) the entire study region and (3) interspecies range overlap. Location: Tropical Africa. Methods: We compiled occurrence data (n = 5,203) on African apes from the IUCN A.P.E.S. database and extracted relevant climate-, habitat- and human-related predictors representing current and future (2050) conditions to predict taxon-specific range change under a best- and a worst-case scenario, using ensemble forecasting. Results The predictive performance of the models varied across taxa. Synergistic interactions between predictors are shaping African ape distribution, particularly human-related variables. On average across taxa, a range decline of 50% is expected outside PAs under the best scenario if no dispersal occurs (61% in worst scenario). Otherwise, an 85% range reduction is predicted to occur across study regions (94% worst). However, range gains are predicted outside PAs if dispersal occurs (52% best, 21% worst), with a slight increase in gains expected across study regions (66% best, 24% worst). Moreover, more than half of range losses and gains are predicted to occur outside PAs where interspecific ranges overlap. Main Conclusions: Massive range decline is expected by 2050, but range gain is uncertain as African apes will not be able to occupy these new areas immediately due to their limited dispersal capacity, migration lag and ecological constraints. Given that most future range changes are predicted outside PAs, Africa's current PA network is likely to be insufficient for preserving suitable habitats and maintaining connected ape populations. Thus, conservation planners urgently need to integrate land use planning and climate change mitigation measures at all decision-making levels both in range countries and abroad