Cathepsin L of Triatoma brasiliensis (Reduviidae, Triatominae): Sequence characterization, expression pattern and zymography

AbstractTriatoma brasiliensis is considered one of the main vectors of Chagas disease commonly found in semi-arid areas of northeastern Brazil. These insects use proteases, such as carboxypeptidase B, aminopeptidases and different cathepsins for blood digestion. In the present study, two genes encoding cathepsin L from the midgut of T. brasiliensis were identified and characterized. Mature T. brasiliensis cathepsin L-like proteinases (TBCATL-1, TBCATL-2) showed a high level of identity to the cathepsin L-like proteinases of other insects, with highest similarity to Rhodnius prolixus. Both cathepsin L transcripts were highly abundant in the posterior midgut region, the main region of the blood digestion. Determination of the pH in the whole intestine of unfed T. brasiliensis revealed alkaline conditions in the anterior midgut region (stomach) and acidic conditions in the posterior midgut region (small intestine). Gelatine in-gel zymography showed the activity of at least four distinct proteinases in the small intestine and the cysteine proteinase inhibitors transepoxysuccinyl-l-leucylamido-(4-guanidino)butane (E-64) and cathepsin B inhibitor and N-(l-3-trans-propylcarbamoyl-oxirane-2-carbonyl)-l-isoleucyl-l-proline (CA-074) were employed to characterize enzymatic activity. E-64 fully inhibited cysteine proteinase activity, whereas in the samples treated with CA-074 residual proteinase activity was detectable. Thus, proteolytic activity could at least partially be ascribed to cathepsin L. Western blot analysis using specific anti cathepsin L antibodies confirmed the presence of cathepsin L in the lumen of the small intestine of the insects

Implementação de Metodologias Ativas em Processos de Educação Corporativa na UFRA / Implementation of Active Methodologies in Corporate Education Processes at UFRA

Objetivou-se com esse trabalho avaliar o impacto das metodologias ativas em processos de Ensino e Aprendizagem, com o objetivo de gerar a autonomia, a coparticipação, a responsabilidade, o protagonismo e a colaboração dos servidores, características que, aliadas, resignificam o ambiente de sala de aula, envolvendo todos os atores em uma perspectiva de Aprendizagem Significativa e centrada no aluno, resultando em uma profunda mudança no modelo formativo atual. Nesse sentido, o relato apresenta o processo de inserção de quatro metodologias ativas em vinte processos de educação corporativa, no âmbito da formação continuada dos servidores da Universidade Federal Rural da Amazônia. Uma vez implementadas as metodologias ativas: Aprendizagem por Pares, Mapas Conceituais, Sala de Aula Invertida e Aprendizagem Baseada em Projetos, a avaliação realizada, os resultados, conforme instrumento de avaliação de reação e avaliação de ensino preenchido por facilitadores e participantes, revelaram que tais metodologias permitiram um maior estímulo à participação, além de contribuírem com uma aprendizagem de forma mais criativa e direcionada à sua atuação profissional

The Genome of Anopheles darlingi, the main neotropical Malaria vector

Anopheles darlingi is the principal neotropical malaria vector, responsible for more than a million cases of malaria per year on the American continent. Anopheles darlingi diverged from the African and Asian malaria vectors ∼100 million years ago (mya) and successfully adapted to the New World environment. Here we present an annotated reference A. darlingi genome, sequenced from a wild population of males and females collected in the Brazilian Amazon. A total of 10 481 predicted protein-coding genes were annotated, 72% of which have their closest counterpart in Anopheles gambiae and 21% have highest similarity with other mosquito species. In spite of a long period of divergent evolution, conserved gene synteny was observed between A. darlingi and A. gambiae. More than 10 million single nucleotide polymorphisms and short indels with potential use as genetic markers were identified. Transposable elements correspond to 2.3% of the A. darlingi genome. Genes associated with hematophagy, immunity and insecticide resistance, directly involved in vector–human and vector–parasite interactions, were identified and discussed. This study represents the first effort to sequence the genome of a neotropical malaria vector, and opens a new window through which we can contemplate the evolutionary history of anopheline mosquitoes. It also provides valuable information that may lead to novel strategies to reduce malaria transmission on the South American continent. The A. darlingi genome is accessible a

Evidence for Reductive Genome Evolution and Lateral Acquisition of Virulence Functions in Two Corynebacterium pseudotuberculosis Strains

Ruiz JC, D'Afonseca V, Silva A, et al. Evidence for Reductive Genome Evolution and Lateral Acquisition of Virulence Functions in Two Corynebacterium pseudotuberculosis Strains. PLoS ONE. 2011;6(4): e18551.Background: Corynebacterium pseudotuberculosis, a Gram-positive, facultative intracellular pathogen, is the etiologic agent of the disease known as caseous lymphadenitis (CL). CL mainly affects small ruminants, such as goats and sheep; it also causes infections in humans, though rarely. This species is distributed worldwide, but it has the most serious economic impact in Oceania, Africa and South America. Although C. pseudotuberculosis causes major health and productivity problems for livestock, little is known about the molecular basis of its pathogenicity. Methodology and Findings: We characterized two C. pseudotuberculosis genomes (Cp1002, isolated from goats; and CpC231, isolated from sheep). Analysis of the predicted genomes showed high similarity in genomic architecture, gene content and genetic order. When C. pseudotuberculosis was compared with other Corynebacterium species, it became evident that this pathogenic species has lost numerous genes, resulting in one of the smallest genomes in the genus. Other differences that could be part of the adaptation to pathogenicity include a lower GC content, of about 52%, and a reduced gene repertoire. The C. pseudotuberculosis genome also includes seven putative pathogenicity islands, which contain several classical virulence factors, including genes for fimbrial subunits, adhesion factors, iron uptake and secreted toxins. Additionally, all of the virulence factors in the islands have characteristics that indicate horizontal transfer. Conclusions: These particular genome characteristics of C. pseudotuberculosis, as well as its acquired virulence factors in pathogenicity islands, provide evidence of its lifestyle and of the pathogenicity pathways used by this pathogen in the infection process. All genomes cited in this study are available in the NCBI Genbank database (http://www.ncbi.nlm.nih.gov/genbank/) under accession numbers CP001809 and CP001829

Tamponamento intrauterino induzido por vácuo para hemorragia pós-parto: uma revisão sistemática

A hemorragia pós-parto (HPP) é uma complicação grave e uma das principais causas de mortalidade materna global, respondendo por aproximadamente 25% de todos os óbitos maternos. A busca por intervenções eficazes e seguras é crítica para melhorar os desfechos maternos. O tamponamento intrauterino induzido por vácuo (VHD) surgiu como uma abordagem promissora, oferecendo potencial para rápido controle do sangramento e redução da necessidade de procedimentos invasivos. Nesse sentido, o presente estudo tem como objetivo analisar a eficácia, segurança e aplicabilidade na prática clínica moderna. Foi realizada uma revisão sistemática da literatura de 2016 a 2024 nas bases de dados PubMed (Medline), Cochrane Library e SciELO. A seleção dos estudos foi baseada em critérios de inclusão e exclusão rigorosos, focando na eficácia, segurança e aplicabilidade do VHD para tratamento da HPP. Três estudos chave foram analisados, com pacientes submetidos ao tratamento com VHD para HPP. Os resultados demonstraram uma taxa de sucesso no tratamento variando de 73% a 94%, com um controle do sangramento alcançado em uma média de 3 minutos. Foi observada uma redução significativa na necessidade de transfusões maciças de sangue e na perda de sangue estimada quando comparado com o tamponamento com balão uterino. Eventos adversos foram relatados, mas todos resolveram-se sem sequelas graves. O tamponamento intrauterino induzido por vácuo apresenta-se como uma opção promissora no tratamento da hemorragia pós-parto, com resultados consistentes indicando eficácia no controle do sangramento e redução na necessidade de transfusões sanguíneas. Embora os resultados sejam encorajadores, mais estudos são necessários para confirmar essas descobertas e explorar plenamente o potencial do VHD na prática clínica. O VHD emerge como uma alternativa eficaz e segura, com potencial para melhorar significativamente os desfechos maternos e reduzir a morbimortalidade associada à HPP

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

The Genome of Anopheles darlingi, the main neotropical malaria vector

Anopheles darlingi is the principal neotropical malaria vector, responsible for more than a million cases of malaria per year on the American continent. Anopheles darlingi diverged from the African and Asian malaria vectors ∼100 million years ago (mya) and successfully adapted to the New World environment. Here we present an annotated reference A. darlingi genome, sequenced from a wild population of males and females collected in the Brazilian Amazon. A total of 10 481 predicted protein-coding genes were annotated, 72% of which have their closest counterpart in Anopheles gambiae and 21% have highest similarity with other mosquito species. In spite of a long period of divergent evolution, conserved gene synteny was observed between A. darlingi and A. gambiae. More than 10 million single nucleotide polymorphisms and short indels with potential use as genetic markers were identified. Transposable elements correspond to 2.3% of the A. darlingi genome. Genes associated with hematophagy, immunity and insecticide resistance, directly involved in vectorhuman and vectorparasite interactions, were identified and discussed. This study represents the first effort to sequence the genome of a neotropical malaria vector, and opens a new window through which we can contemplate the evolutionary history of anopheline mosquitoes. It also provides valuable information that may lead to novel strategies to reduce malaria transmission on the South American continent. The A. darlingi genome is accessible at www.labinfo.lncc.br/index.php/anopheles- darlingi. © 2013 The Author(s)

Early Growth and Neurologic Outcomes of Infants with Probable Congenital Zika Virus Syndrome

We report the early growth and neurologic findings of 48 infants in Brazil diagnosed with probable congenital Zika virus syndrome and followed to age 1–8 months. Most of these infants had microcephaly (86.7%) and craniofacial disproportion (95.8%). The clinical pattern included poor head growth with increasingly negative z-scores, pyramidal/extrapyramidal symptoms, and epilepsy