Implementation of a Practical Teaching Course on Protein Engineering

Simple Summary Proteins are the workhorses of the cell. With different combinations of the 20 common amino acids and some modifications of these amino acids, proteins have evolved with a staggering array of new functions and capabilities due to Protein Engineering techniques. The practical course presented was offered to undergraduate bioengineering and chemical students at the Faculty of Engineering of the University of Porto (Portugal) and consists of sequential laboratory sessions to learn the basic skills related to the expression and purification of recombinant proteins in bacterial hosts. These experiments were successfully applied by students as all working groups were able to isolate a model recombinant protein (the enhanced green fluorescent protein) from a cell lysate containing a mixture of proteins and other biomolecules produced by an Escherichia coli strain and evaluate the performance of the extraction and purification procedures they learned. Protein Engineering is a highly evolved field of engineering aimed at developing proteins for specific industrial, medical, and research applications. Here, we present a practical teaching course to demonstrate fundamental techniques used to express, purify and analyze a recombinant protein produced in Escherichia coli-the enhanced green fluorescent protein (eGFP). The methodologies used for eGFP production were introduced sequentially over six laboratory sessions and included (i) bacterial growth, (ii) sonication (for cell lysis), (iii) affinity chromatography and dialysis (for eGFP purification), (iv) bicinchoninic acid (BCA) and fluorometry assays for total protein and eGFP quantification, respectively, and (v) sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for qualitative analysis. All groups were able to isolate the eGFP from the cell lysate with purity levels up to 72%. Additionally, a mass balance analysis performed by the students showed that eGFP yields up to 46% were achieved at the end of the purification process following the adopted procedures. A sensitivity analysis was performed to pinpoint the most critical steps of the downstream processing

Três miradas sobre o Mangue: representações da prostituição carioca na primeira metade do século XX

Em um momento em que o Rio de Janeiro se modernizava, com as reformas urbanas, o Mangue, zona de baixo meretrício carioca, era o emblema máximo da sujeira. O espaço representava tudo aquilo que as classes dominantes desejavam ignorar em prol de um pretenso desenvolvimento e de uma alegada higienização urbana. Jornalistas, juízes, médicos e demais pessoas que ocupavam posições de poder na sociedade tinham particular interesse pela prostituição, tema percebido na época como intrinsecamente ligado ao comportamento da mulher e à moralidade pública, em uma época de intenso crescimento urbano, conforme aponta Margareth Rago (2008, p. 21). Considerando o exercício da prostituição uma atividade remunerada, e levando em consideração também o contextohistórico, o presente artigo busca analisar como as mulheres que vendiam serviços sexuais eram representadas nas artes e na literatura. Para isso, escolhemos três discursos sobre o assunto: o poema “O santeiro do Mangue”, de Oswald de Andrade; as gravuras que compõem o álbum Mangue, de Lasar Segall e algumas obras (sem título) produzidas por Aurora Cursino, pintora que trabalhou como prostituta no Rio de Janeiro durante as décadas de 1920 e 1930.At a time when Rio de Janeiro was modernizing itself, with urban reforms, the Mangue, Rio's low red-light district, was the ultimate emblem of dirt. The Mangue represented everything upper classes wanted to ignore in favor of an alleged urban development and sanitation. Journalists, judges, doctors and other people who occupied positions of power in society were particularly interested in prostitution, a topic perceived at the time as intrinsically linked to women's behavior and public morality, in a time of intense urban growth, as pointed out by Margareth Rago (2008, p. 21) Considering the exercise of prostitution as a paid activity, and also taking into consideration the context of the epoch, this article seeks to analyze how women who sold sexual services were represented in the arts and literature. Thus, we chose three speeches about the subject: the poem “O santeiro do Mangue”, by Oswald de Andrade; the engravings that make up the album Mangue, by Lasar Segall and some works (untitled) produced by Aurora Cursino, a painter who also worked as a prostitute in Rio de Janeiro during the 1920s and 1930s

Formulação e memória na trama discursiva da antologia de ensaios Nenhum Brasil existe: Antônio Cândido como discurso fundador na construção da memória crítico-literária nacional

RESUMO: Tendo como ponto de partida a organização discursiva da antologia de ensaios Nenhum Brasil existe, organizada por João Cezar de Castro Rocha, buscarei examinar as marcas de heterogeneidade no tocante à voz de Antonio Candido, no ensaio introdutório da antologia. Para tanto, levei em conta o conceito de discurso fundador (ORLANDI, 2003), na medida em que os dizeres fundadores de Candido na ensaística brasileira se constituem como processos de produção de sentidos que possibilitam o surgimento de outros processos discursivos em relação ao Brasil e à identidade nacional, na tensão entre repetição e deslocamento

Association of increased levels of homocysteine and peripheral arterial disease in a Japanese-Brazilian population

Objectives. Our aim was to evaluate the possible association between homocysteine levels and peripheral arterial disease (PAD) in a population-based study of Japanese-Brazilians.Materials and methods. This cross-sectional study was derived from a population-based survey on the prevalence of diabetes and associated diseases conducted in Japanese-Brazilians. A total of 1330 male and female subjects aged >= 30 years were submitted to clinical examination and laboratory procedures including homocysteine measurement. the ankle-brachial index (ABI) was calculated, subjects with ABI values <0.9 were diagnosed with PAD. the evaluable population included 1008 subjects. Logistic regression was used taking PAD as the dependent variable.Results. Mean age of the population was 56.5 years and overall prevalence of PAD was 20%. A worse cardiovascular profile was found in male patients, including significantly higher homocysteine levels (11.9 +/- 1.8 vs. 9.1 +/- 1.1 mu mol/L, p < 0.001). Men with PAD had higher prevalence rates of hyperhomocysteinemia compared to women (22.7% vs 7.6%). Univariate analysis showed an odds ratio of hyperhomocysteinemia for PAD of 1.51 [1.02-2.25] in men and 1. 69 [1.06-2.68] in women. After adjustment for other cardiovascular risk factors, higher levels of homocysteine were only significantly related to PAD in men.Conclusion. in a Japanese-Brazilian population, elevated levels of homocysteine are associated with PAD in men. Prospective studies are necessary to confirm this finding.Universidade Federal de São Paulo, Dept Surg, Postgrad Course Cardiac Vasc & Thorac Surg, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Surg, Div Vasc Surg, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Pediat, São Paulo, BrazilUniv São Paulo, Sch Publ Hlth, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Surg, Postgrad Course Cardiac Vasc & Thorac Surg, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Surg, Div Vasc Surg, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Pediat, São Paulo, BrazilWeb of Scienc

O ensino da leitura em língua materna ancorado na perspectiva discursiva: uma proposta de leitura de um conto contemporâneo

Considering the importance of thinking the foundations of reading teaching in mother tongue, this paper presents a didactic approach reading discursive, keeping in view the mobilization of concepts, namely: production conditions, linguistic surface and senses of incorporation processes (in relation to memory), based on a short story published in “O Globo” newspaper that circulates in a literary blog named Prosa. The proposal aims to present possible referrals to the text and discuss some issues related to the mother tongue of reading instruction such as evidence of the senses, the contradiction and the equivocity.Keywords: reading concepts, reading in mother tongue, Discourse Analysis.Considerando a relevância de pensar as bases do ensino da leitura de língua materna, neste artigo, apresentaremos uma abordagem didática de leitura de cunho discursivo, tendo-se em vista a mobilização dos conceitos, a saber: condições de produção, superfície linguística e processos de constituição de sentidos (na relação com a memória), com base em um conto publicado no sítio eletrônico do jornal O Globo intitulado “O Corpo” que circula em um blog literário do referido jornal chamado Prosa. A proposta tem como objetivo apresentar possíveis encaminhamentos para o texto e problematizar algumas questões pertinentes ao ensino da leitura de língua materna, tais como a evidência dos sentidos, a contradição e a equivocidade.Palavras-chave: concepções de leitura, leitura em língua materna, Análise do Discurso

O ensino da leitura em língua materna ancorado na perspectiva discursiva: uma proposta de leitura de um conto contemporâneo

Considerando a relevância de pensar as bases do ensino da leitura de língua materna, neste artigo, apresentaremos uma abordagem didática de leitura de cunho discursivo, tendo-se em vista a mobilização dos conceitos, a saber: condições de produção, superfície linguística e processos de constituição de sentidos (na relação com a memória), com base em um conto publicado no sítio eletrônico do jornal O Globo intitulado “O Corpo” que circula em um blog literário do referido jornal chamado Prosa. A proposta tem como objetivo apresentar possíveis encaminhamentos para o texto e problematizar algumas questões pertinentes ao ensino da leitura de língua materna, tais como a evidência dos sentidos, a contradição e a equivocidade.Palavras-chave: concepções de leitura, leitura em língua materna, Análise do Discurso.</p

Coronavirus Disease 2019 and Human Reproduction: A Changing Perspective

Since the outbreak of severe acute respiratory coronavirus 2 (SARS-CoV-2), the coronavirus disease 2019 has had a wide range of effects on human health. This paper summarizes the data related to the effects of the SARS-CoV-2 infection on human reproduction. Both the male and female reproductive tract express high levels of receptors and proteins needed for viral cell entry. There is presently no evidence that gametes are affected by the infection. Male fertility may be temporarily reduced due to inflammatory responses following infection. The endometrium is highly susceptible to SARS-CoV-2 cell entry; however, it remains unclear whether this could alter receptivity and embryo implantation. Menstrual cycle changes were reported in women who experienced severe infection; however, they tended to be reversible. For couples undergoing assisted reproduction treatment, the pandemic led to a significant psychological burden, with changes in lifestyle that could directly affect the success of the treatment. Human reproduction societies recommend screening all patients prior to cycle initiation and avoiding treatment of women with severe comorbidities until the pandemic is under control. Finally, for pregnant women, it is expected that the infection is more severe in women in the third trimester and in those with comorbidities. Those who are symptomatic for SARS-CoV-2 are more likely to have increased rates of prematurity and intrapartum fetal distress than those who are asymptomatic. Vertical transmission cannot be completely ruled out, but neonatal infection rates are low. Vaccination appears to be safe and is indicated for use in pregnant and lactating women because the benefits outweigh the risks