Determinação da expressão da arginase 1 e sintase do óxido nítrico induzível durante a gravidez e infeção por toxoplasma gondii

Mestrado em Biologia Molecular e CelularToxoplasma gondii is considered to be the world’s most successful zoonotic parasite, causing congenital toxoplasmosis, a prevalent disease worldwide with serious implications for the fetus. The mechanisms playing a role in the induction of pathology during infection are not clear, but are potentially associated with disruption of normal homeostatic immunological pathways including macrophage activation. Macrophages have a variety of activation states, being able to adapt their functions to environmental changes of cytokines, described as innate (TLR ligation), classical/M1 (TLR ligation with IFN) or alternative/M2 (IL4Ralpha ligation) activation states. Thus, while classical macrophage activation can control replication through induction of inducible nitric oxide synthase (NOS2), alternative activation can control parasite replication through induction of arginase 1 (Arg-1) and depletion of arginine. Our study was focused on the effects of T. gondii infection on Arg-1 and NOS2 expression at the fetomaternal interface (FMI) and systemic level (peritoneal exudate cells and spleen cells) using the mice model. Infection of pregnant BALB/c and C57Bl/6 (B6) mice with a type II strain of T. gondii allowed the follow-up of pregnancy. Morphometric analysis of decidua and placenta was performed using haematoxylin-eosin sections of the fetoplacental units. The parasite loads’ evaluation was done by quantitative Real Time-PCR (q-PCR) using Taqman probes. Arg-1 and NOS2 expression were evaluated by qRT-PCR, immunohistochemistry and Western Blotting. It was observed a significant decrease in placental and decidual areas in infected B6 compared to control but not in Balb/c mice. Immunohistochemical analysis indicated an increased expression of Arg-1 and a decreased expression of NOS2 in the decidua from infected compared to control animals from both strains of mice. This indicates that infection interferes with the process of placentation delaying decidualization and this might be correlated with altered expression of Arg-1 and NOS2 in B6 but not in Balb/c mice. This work aims not only understanding the role of macrophage activation in congenital infection by T. gondii, but also should provide valuable information regarding the role of macrophages during healthy pregnancy and infection complicated pregnancy.Toxoplasma gondii é considerado um parasita zoonótico, causando toxoplasmose congênita, uma doença prevalente em todo o mundo com implicações graves para o feto. Os mecanismos que desempenham um papel, induzindo patologia durante a infeção por este parasita não são claros, mas são potencialmente associados com alterações dos processos homeostáticos imunológicos normais, incluindo a ativação de macrófagos. Os macrófagos apresentam uma variedade de estadios de ativação descritos como ativação inata (ligação TLR), clássica/M1 (ligação TLR com IFN) ou alternativa/M2 (ligação IL4Ralpha). Assim, enquanto os macrófagos M1 controlam a replicação dos parasitas pela indução de sintetase do óxido nítrico induzível (NOS2), os macrófagos M2 controlam a replicação dos parasitas induzindo a arginase-1 (Arg-1) e depletando a arginina. O nosso estudo incidiu sobre os efeitos da infeção por T. gondii, na expressão de Arg-1 e de NOS2 na interface feto-materna (FMI) e a nível sistémico (células de exsudado peritoneal e células do baço) utilizando murganhos como modelo de estudo. A infeção de animais gestantes Balb/c e C57BL/6 (B6) com uma estirpe de T. gondii tipo II permitiu o acompanhamento da gravidez. A análise morfométrica da decídua e da placenta foi realizada utilizando seções de unidades fetoplacentárias coradas com hematoxilina-eosina. A determinação da carga parasitária foi realizada por PCR quantitativo em tempo real utilizando sondas Taqman (q-PCR). A expressão de Arg-1 e NOS2 foi avaliada por qRT-PCR, Imuno-histoquímica e Western-Blotting. Observou-se uma diminuição significativa na área da placenta e da decídua em B6 infetados em comparação com os animais controlo, mas não nos animais Balb/c. A análise imunohistoquímica indicou um aumento da expressão de Arg-1 e uma diminuição da expressão de NOS2 na decídua de animais infetados em comparação com os animais controlo em ambas as estirpes. Os resultados obtidos sugerem que a infeção interfere com o processo de placentação atrasando a decidualização, podendo existir uma relação com as alterações da expressão de Arg-1 e NOS2 nos B6, mas não nos animais Balb/c. O trabalho desenvolvido visa não só a compreensão do papel da ativação de macrófagos na infeção congénita pelo T. gondii, mas poderá fornecer informações essenciais sobre o papel dos macrófagos durante uma gravidez saudável e uma gravidez com infeção

Electroactive polymer membranes as substrates for point-of-care devices

Point-of-care devices (POC) are becoming essential for medical assistance in emergency situations or location with difficult access to medical infrastructures. In this work, innovative microfluidic substrates based on electroactive poly(vinylidene-co-trifluorethylene)-P(VDF-TrFE) with tuned morphologies and adequate physicochemical properties were developed using electrospinning and phase inversion techniques, as alternative to commercially available two-dimensional microfluidic substrates based mainly on cellulose. Their hydrophilicity was tuned using plasma treatments and barriers were implemented using wax printing to fabricate a design able to carry out glucose assays as a proof of concept.FCT- Fundação para a Ciência e Tecnologia: PTDC/EMD-EMD/28159/2017; SFRH/BD/140698/2018. Spanish Ministry of Economy and Competitiveness (MINECO): MAT2016-76039-C4-3-R. Basque Government Industry and Education Departments: ELKARTEK, HAZITEK and PIBA (PIBA-2018-06

Natural based reusable materials for microfluidic substrates: The silk road towards sustainable portable analytical systems

Portable analytical systems are versatile tools for application in areas including biomedicine, biosecurity, food safety and environmental monitoring. This work contributes to the increasing demand for low-cost, environmentally friendly substrates for portable analytical systems by using natural Bombyx mori cocoons. Further, silk fibroin is also extracted from these cocoons and electrospun into oriented and randomly oriented fiber substrates. Oxygen plasma treatment is applied to improve their hydrophilicity. Fiber morphology, mechanical properties, porosity, thermal characteristics and surface contact angle are extensively characterized and the ability of the samples for passive capillary flows demonstrated. Plasma treated pressed cocoons show superhydrophilicity, capillary flow rates of 44.8 ± 3.75 mm.min-1, and high mechanical resistance with Young's modulus values up to 592.13 ± 19.83 MPa. The developed materials are used as substrates for the colorimetric quantification of three commonly scrutinized clinical analytes. Hydrophobic barriers are first wax-printed on all samples with a proper design and albumin assays are performed on all substrates. Further assays for uric acid and glucose quantification are successfully accomplished on the pressed cocoons after a simple in between washing step, with overall high coefficient of determination, proving the suitability of the developed materials as low-cost, sustainable and reusable microfluidic substrates.This work was supported by the Portuguese Foundation for Science and Technology (FCT) under strategic funding UID/FIS/04650/2020, UIDB/04436/2020, UIDP/04436/2020 and project PTDC/EMD-EMD/28159/2017 (POCI-01–0145-FEDER-028159). The authors also thank FCT for financial support under grants SFRH/BD/140698/2018 (R.B. P.), 2020.09218.BD (A.S.M.), 2020.04163.CEECIND (C.R.) and 2020.02304.CEECIND (V.F.C.). Finally, the authors acknowledge fund ing by Spanish State Research Agency (AEI) and the European Regional Development Fund (ERFD) through the project PID2019–106099RB C43/AEI/10.13039/501100011033 and from the Basque Government Industry Departments under the ELKARTEK program. Finally, the au thors also thank Dr. J. Borges and Prof. F. Vaz for experimental support.info:eu-repo/semantics/publishedVersio

Specific probiotics in elite athlete injuries: indication for usage / Probiotics específicos em lesões de atletas elite: indicação de uso

More severe effects of illness that require probiotic intervention and complete cessation of training activities might have a larger effect on competitive sport performance. In this review, we outline key principles of prevention for injury and illness in high level athletes using probiotic treatments, examine relevant scientific evidence on probiotics for reducing the risk of illness and injury, and summarize practical recommendations for high level athletes, coaches, and practitioners. Studies were obtained through manual and electronic journal searches, review articles, as well as personal correspondence, concerning the period 2000-2018. We used the following databases: PubMed, Web of Science, SCOPUS and Medline. Electronic databases were searched using keywords and/or MeSH terms such as [[[Athlete] or [Sport effects] or [Injuries] or [Disease] or [Allergy] or [Illness] or [Sickness] or [Desorder]] in combination with [[microbiota] or [microbiotas] or [Microbiome] or [microbiomes] or [Human Microbiome] or [Human microbiomes] or [microbiomes, Human] or [Microbiome, Human]] not [[disease]]], until February 18th, 2018. In this present review, we have outlined key points of injury and illness prevention for elite athletes using probiotic treatments, examining relevant scientific evidence on probiotics for reducing the risk of illness and injury, and summarized practical recommendations for high level athletes, coaches, and practitioners

Análise psicofisiológica em simulação de combate competitivo de artes marciais mistas / Psychophysiological analysis in competitive combat simulation of mixed martial arts

Este estudo comparou o estado de humor e variáveis fisiológicas de combates de MMA, antes e depois de uma simulação de competição. A amostra foi composta por 10 atletas em 3 rounds consecutivos de lutas masculinas em medidas agudas pré e pós-combate. O estado de humor foi medido pelo questionário BRUMS, a percepção subjetiva de esforço (PSE) foi medida pela escala de Borg (0-10). Foram verificadas pressão arterial diastólica e sistólica, frequência cardíaca, concentração de lactato, testosterona e cortisol. Todos os dados foram normais e comparados de maneira pareada pelo test t de Student, com p≤0,05. A comparação entre os valores pré e pós mostraram diferenças em tensão (7,1±6,43,6±3,8 pontos), pressão arterial sistólica (11,9±0,813,3±1,5 mmHg), frequência cardíaca (75±33152,7±63,6 bpm), concentração de lactato (2,5±1,010±1,8 u/L), PSE (0,9±13±1,1 pontos), cortisol (95,1±53,6125,0±59,4 ng/dL) e testosterona (3,4±0,44,4±0,4 ng/dL). Por sua vez, a análise de regressão mostrou que alterações no estado de humor dos atletas têm relação com a pressão arterial diastólica. A tensão e a fadiga dos atletas influenciaram a pressão arterial diastólica e, consequentemente, a vasodilatação o que pode desfavorecer o fornecimento de oxigênio para a realização das ações necessárias durante os treinamentos e combates de MMA

Prematuridade em gestações resultantes de fertilização in vitro

A quantitative, descriptive, retrospective and documentary study is presented to assess the rate of premature births in pregnancies resulting from in vitro fertilization, using the Cryolife program for data collection, which is the program used by the clinic present in the research, for storage of patients' electronic medical records. The study was carried out in a human reproduction clinic in northern Paraná, where we established criteria to identify the index and possible causes of premature births in women with a prevalence of age from 34 to 38 years old who became pregnant through in vitro fertilization among the years 2015 and 2019, using the fresh embryo transfer technique or embryo thaw. It was obtained that in this period 156 babies were born, 59.61% were male, 64.1% were single gestation, 23% were born less than 37 weeks old and 33.3% were born weighing less than 2500 g. And relating to single and twin pregnancies, it was found that 50% of premature babies and 71.15% of babies born with low weight were twins. It is concluded that the rate of premature births in IVF is high, however there was a predominance of single pregnancies during the human reproduction process in the 4 years evaluated.Apresenta-se um estudo quantitativo, descritivo, retrospectivo e documental para avaliação do índice de partos prematuros em gestações resultantes de fertilização in vitro, mediante a utilização do programa Cryolife para coleta de dados, sendo este o programa utilizado pela clínica presente na pesquisa, para armazenamento dos prontuários eletrônicos das pacientes. O estudo foi realizado em uma clínica de reprodução humana do norte do Paraná, onde estabelecemos critérios para identificar o índice e possíveis causas dos partos prematuros em mulheres com a prevalência de idade de 34 a 38 anos que engravidaram por meio da fertilização in vitro entre os anos de 2015 e 2019, utilizando a técnica de transferência de embrião a fresco ou descongelamento de embrião. Obteve-se que neste período nasceram 156 bebês, 59,61% eram do sexo masculino, 64,1% eram de gestação única, 23% nasceram de menos de 37 semanas e 33,3% nasceram com peso inferior 2500 g. E relacionando com gestações únicas e gemelares, obteve-se que 50% dos prematuros e 71,15% dos bebês que nasceram com baixo peso eram gêmeos. Conclui-se que o índice de partos prematuro na fertilização in vitro é realmente elevado, contudo houve a predominância de gestações únicas durante do processo de reprodução humana nos 4 anos avaliados

Challenges of public education in Brazil: Notes on school, the family and the community

This paper proposed to delimit as an object of study the “Challenges faced by Public Education in Brazil”, without the purpose of verifying the quality of education in the national territory. We understand that it is necessary for the school, the family and the community to walk together, so that socio-educational quality can be achieved. Therefore, an analysis of the main productions available on the subject is necessary. Although access to Elementary Education is practically universal, Brazil still faces the great problem of the precarious quality of education offered in its schools. Thus, the general objective of this work is to analyze the strategies that can be used by the school, the family and the community to implement the qualification of public education, specifically in Paraíba. Thus, subsidies to exercise and studies on the subject arise. This work is bibliographic and documental, under a qualitative approach, since it is understood as a social quality strategy and legal regulations through documents and bibliographic materials related to the strategies can be used by the school, family and community, to carry out the qualification of the public education, in Paraíba