Study of the antimalarial properties of hydroxyethylamine derivatives using green fluorescent protein transformed Plasmodium berghei.

A rapid decrease in parasitaemia remains the major goal for new antimalarial drugs and thus, in vivo models must provide precise results concerning parasitaemia modulation. Hydroxyethylamine comprise an important group of alkanolamine compounds that exhibit pharmacological properties as proteases inhibitors that has already been proposed as a new class of antimalarial drugs. Herein, it was tested the antimalarial property of new nine different hydroxyethylamine derivatives using the green fluorescent protein (GFP)-expressing Plasmodium berghei strain. By comparing flow cytometry and microscopic analysis to evaluate parasitaemia recrudescence, it was observed that flow cytometry was a more sensitive methodology. The nine hydroxyethylamine derivatives were obtained by inserting one of the following radical in the para position: H, 4Cl, 4-Br, 4-F, 4-CH3, 4-OCH3, 4-NO2, 4-NH2 and 3-Br. The antimalarial test showed that the compound that received the methyl group (4-CH3) inhibited 70% of parasite growth. Our results suggest that GFP-transfected P. berghei is a useful tool to study the recrudescence of novel antimalarial drugs through parasitaemia examination by flow cytometry. Furthermore, it was demonstrated that the insertion of a methyl group at the para position of the sulfonamide ring appears to be critical for the antimalarial activity of this class of compounds.

Heme on Pulmonary Malaria: Friend or Foe?

Malaria is a hemolytic disease that, in severe cases, can compromise multiple organs. Pulmonary distress is a common symptom observed in severe malaria caused by Plasmodium vivax or Plasmodium falciparum. However, biological components involved in the development of lung malaria are poorly studied. In experimental models of pulmonary malaria, it was observed that parasitized red blood cell-congested pulmonary capillaries are related to intra-alveolar hemorrhages and inflammatory cell infiltration. Thus, it is very likely that hemolysis participates in malaria-induced acute lung injury. During malaria, heme assumes different biochemical structures such as hemin and hemozoin (biocrystallized structure of heme inside Plasmodium sp.). Each heme-derived structure triggers a different biological effect: on the one hand, hemozoin found in lung tissue is responsible for the infiltration of inflammatory cells and consequent tissue injury; on the other hand, heme stimulates heme oxygenase-1 (HO-1) expression and CO production, which protect mice from severe malaria. In this review, we discuss the biological mechanism involved in the dual role of heme response in experimental malaria-induced acute lung injury.

Urgência da geração de conhecimento durante a pandemia de covid-19: um retrospecto sobre a integridade em publicações em saúde / Urgency of knowledge generation during the covid-19 pandemic: a retrospective on health publication integrity / Urgencia de generación de conocimiento durante la pandemia de covid-19: una retrospectiva a la integridad en las publicaciones sobre salud

Durante a pandemia de covid-19, foi observado um aumento expressivo do número de publicações (artigos e preprints), que levou ao rápido compartilhamento de informações e incentivou a discussão sobre a integridade científica na geração do conhecimento durante emergências sanitárias. Nesse sentido, o objetivo deste trabalho foi o de realizar uma breve análise do cenário referente à integridade em pesquisa em publicações em saúde durante a pandemia de covid-19. Para isso, fizemos uma pesquisa documental em sites de organizações com histórico de promoção da cultura da integridade. Verificamos como a urgência de geração de conhecimento acelerou, de forma positiva, o debate sobre ética e integridade em pesquisa e ciência aberta. Por outro lado, esse contexto expôs pontos críticos, como práticas questionáveis e/ou fraude em pesquisa. Ações educativas em instituições de pesquisa que visem à implementação e à manutenção da cultura da integridade podem contribuir significativamente para transformações positivas no sistema de pesquisa

During the covid-19 pandemic, a significant increase in the number of publications (articles and preprints) was observed, which led to the rapid sharing of information and encouraged the discussion about scientific integrity in the generation of knowledge during health emergencies. In this sense, the present work aims to analyze research integrity in health publications during the covid-19 pandemic. For this goal, we conducted documentary research on websites of organizations that promote the culture of research integrity. We verified how the urgency of generating knowledge positively accelerated the debate on ethics and integrity in research and open science. On the other hand, this context exposed critical points, such as questionable research practices, and/or research fraud. Educational actions in research institutions aimed at implementing and maintaining a culture of integrity can significantly contribute to positive changes in the research system.

Durante la pandemia de covid-19, se observó un aumento en el número de publicaciones (artículos y preprints), lo que condujo al rápido intercambio de información y fomentó la discusión sobre la integridad científica en la generación de conocimiento durante las emergencias sanitarias. El objetivo de este trabajo fue realizar un análisis sobre la integridad de la investigación en publicaciones de salud durante la pandemia de covid-19. Realizamos una investigación documental en sitios web de organizaciones con trayectoria en la promoción de una cultura de integridad. Comprobamos cómo la urgencia de generar conocimiento aceleró positivamente el debate sobre ética e integridad en la investigación y la ciencia abierta, pero expuso puntos críticos, tales como prácticas cuestionables y/o fraude de investigación. Las acciones educativas en instituciones de investigación dirigidas a implementar y mantener una cultura de integridad pueden contribuir a cambios positivos en el sistema de investigación.

Angiotensin II is a new component involved in splenic T lymphocyte responses during Plasmodium berghei ANKA infection.

The contribution of T cells in severe malaria pathogenesis has been described. Here, we provide evidence for the potential role of angiotensin II (Ang II) in modulating splenic T cell responses in a rodent model of cerebral malaria. T cell activation induced by infection, determined by 3 to 4-fold enhancement in CD69 expression, was reduced to control levels when mice were treated with 20 mg/kg losartan (IC50 = 0.966 mg/kg/d), an AT1 receptor antagonist, or captopril (IC50 = 1.940 mg/kg/d), an inhibitor of angiotensin-converting enzyme (ACE). Moreover, the production of interferon-γ and interleukin-17 by CD4+ T cells diminished 67% and 70%, respectively, by both treatments. Losartan reduced perforin expression in CD8+ T cells by 33% while captopril completely blocked it. The upregulation in chemokine receptor expression (CCR2 and CCR5) observed during infection was abolished and CD11a expression was partially reduced when mice were treated with drugs. T cells activated by Plasmodium berghei ANKA antigens showed 6-fold enhance in AT1 levels in comparison with naive cells. The upregulation of AT1 expression was reduced by losartan (80%) but not by captopril. Our results suggest that the AT1/Ang II axis has a role in the establishment of an efficient T cell response in the spleen and therefore could participate in a misbalanced parasite-induced T cell immune response during P. berghei ANKA infection.

Synthesis and in vivo antimalarial evaluation of novel hydroxyethylamine derivatives.

A series of hydroxyethylamines has been synthesized from the reaction of (2S,3S )Boc-phenylalanine epoxide with alkyl amines in good yields and evaluated for their in vivo antimalarial activity in mice. Compound 4g presented better activity then the reference artesunate in percentage of inhibition of parasitemia in treated P. berghei-infected mice and compare to the activity of artesunate in the survival of mice 14 days after infection. In addiction, no hemolytic activity was found, which supports that inhibition of parasitemia is due to antimalarial activity. The compound 4g inhibited the differentiation to schizonts suggesting that parasite metabolism is a possible target of 4g. These results indicate that this class of compound possesses promising perspectives for the development of new antimalarial drugs.

AT1 receptor-mediated angiotensin II activation and chemotaxis of T lymphocytes.

Angiotensin II (Ang II), a central renin-angiotensin system (RAS) effector molecule, and its receptors, AT(1) and AT(2), have been shown to be involved in the inflammatory aspects of different diseases, however the cellular mechanisms underlying the regulation of immunity are not fully understood. In this work, using spleen-derived CD4(+) and CD8(+) T lymphocytes activated in vitro, we tested the influence of Ang II on different aspects of the T cell function, such as activation and adhesion/transmigration through endothelial basal membrane proteins. The addition of 10(-8)M Ang II did not change any of the parameters evaluated. However, 10(-6)M losartan, an AT(1) receptor antagonist: (i) reduced the percentage of CD25(+) and CD69(+) cells of both subsets; (ii) inhibited adhesion of these cells to fibronectin or laminin by 53% or 76%, respectively and (iii) significantly reduced transmigration through fibronectin or laminin by 57% or 43%, respectively. In addition, 10(-6)M captopril, an angiotensin-converting enzyme inhibitor had similar effects to Ang II, however its effects were reverted by exogenous Ang II (10(-8)M). None of these responses was modified by 10(-7)M PD123319, an AT(2) antagonist. These data reinforce the notion of endogenous production of Ang II by T cells, which is important for T cell activation, and adhesion/transmigration induced on interaction with basal membrane proteins, possibly involving AT(1) receptor activation. Moreover, AT(1) receptor expression is 10-fold higher in activated T lymphocytes compared with naive cells, but AT(2) receptor expression did not change after T cell receptor triggering.

Impairment of the Plasmodium falciparum erythrocytic cycle induced by angiotensin peptides.

Plasmodium falciparum causes the most serious complications of malaria and is a public health problem worldwide with over 2 million deaths each year. The erythrocyte invasion mechanisms by Plasmodium sp. have been well described, however the physiological aspects involving host components in this process are still poorly understood. Here, we provide evidence for the role of renin-angiotensin system (RAS) components in reducing erythrocyte invasion by P. falciparum. Angiotensin II (Ang II) reduced erythrocyte invasion in an enriched schizont culture of P. falciparum in a dose-dependent manner. Using mass spectroscopy, we showed that Ang II was metabolized by erythrocytes to Ang IV and Ang-(1-7). Parasite infection decreased Ang-(1-7) and completely abolished Ang IV formation. Similar to Ang II, Ang-(1-7) decreased the level of infection in an A779 (specific antagonist of Ang-(1-7) receptor, MAS)-sensitive manner. 10(-7) M PD123319, an AT(2) receptor antagonist, partially reversed the effects of Ang-(1-7) and Ang II. However, 10(-6) M losartan, an antagonist of the AT(1) receptor, had no effect. Gs protein is a crucial player in the Plasmodium falciparum blood cycle and angiotensin peptides can modulate protein kinase A (PKA) activity; 10(-8) M Ang II or 10(-8) M Ang-(1-7) inhibited this activity in erythrocytes by 60% and this effect was reversed by 10(-7) M A779. 10(-6) M dibutyryl-cAMP increased the level of infection and 10(-7) M PKA inhibitor decreased the level of infection by 30%. These results indicate that the effect of Ang-(1-7) on P. falciparum blood stage involves a MAS-mediated PKA inhibition. Our results indicate a crucial role for Ang II conversion into Ang-(1-7) in controlling the erythrocytic cycle of the malaria parasite, adding new functions to peptides initially described to be involved in the regulation of vascular tonus.

Phytosociology of weeds in millet under different soil managements in savanna sul-mato-grossense / Fitossociologia de plantas daninhas em milheto sob diferentes manejos do solo no cerrado sul-mato-grossense

The millet crop in recent decades has shown an increase in planted area, mainly in the Cerrado region. However, there are few studies related to the management and phytossociology of weeds in this culture. Thus, the objective of this research was to perform a phytosociological survey in millet under different soil managements in Savanna Sul-Mato-Grossense region. The experiment was conducted in the experimental area of Universidade Estadual de Mato Grosso do Sul ­ Unit of Aquidauana (UEMS/UUA), in Aquidauana-MS. The area is cultivated to five years with soybeans in first crop and corn in second. The experimental design was randomized complete block in a split-plot design consisting of four blocks with four replications. In the plots were used soil preparation systems (minimum tillage and no-tillage) and in the subplots was utilized the nitrogen fertilization at 50 kg ha-1 and absence of nitrogen on millet at 25 days after emergence (DAE). Were evaluated the parameters number of species (NS), total number and dry mass of weeds (TN and DMW, respectively), dry mass of millet (DMM) and frequency (F), density (D), abundance (A) and Importance Value (IV) of weeds. The no-tillage system provided greater dry mass of weeds, whereas the nitrogen resulted in a smaller total number of weeds (TN). Cynodon dactylon and Commelina benghalensis were the most predominant species on area.

O cultivo do milheto vem apresentando nas últimas décadas um crescimento da área plantada, sobretudo na região do Cerrado. Entretanto, são escassos os estudos relacionados ao manejo e à fitossociologia de plantas daninhas nesta cultura. Diante disto, o objetivo desta pesquisa foi realizar um levantamento fitossociológico em milheto sob diferentes manejos do solo na região do Cerrado sul-mato-grossense. O experimento foi instalado no setor de Fitotecnia da Universidade Estadual de Mato Grosso do Sul, Unidade Universitária de Aquidauana (UEMS/UUA), município de Aquidauana-MS. A área é cultivada a cinco anos com soja em primeira safra e milho em segunda. O delineamento experimental utilizado foi o de blocos casualizados em esquema de parcelas subdivididas, constituído por quatro blocos e quatro repetições. Nas parcelas foram avaliados os sistemas de preparo do solo (cultivo mínimo e plantio direto) e nas subparcelas a adubação nitrogenada de 50 kg ha-1 e a ausência de nitrogênio em cobertura no milheto aos 25 dias após a emergência (DAE). Foram avaliados os parâmetros número de espécies (NE), massa seca e número total de plantas daninhas (MSPD e NT, respectivamente), massa seca do milheto (MSM) e frequência (F), densidade (D), abundância (A) e Valor de Importância (IV) das espécies de plantas daninhas. O sistema de plantio direto proporcionou uma maior massa seca de plantas daninhas, enquanto que a adubação nitrogenada conferiu um menor número total de plantas invasoras. Cynodon dactylon e Commelina benghalensis foram as espécies mais predominantes na área.

Study of the antimalarial properties of hydroxyethylamine derivatives using green fluorescent protein transformed Plasmodium berghei

A rapid decrease in parasitaemia remains the major goal for new antimalarial drugs and thus, in vivo models must provide precise results concerning parasitaemia modulation. Hydroxyethylamine comprise an important group of alkanolamine compounds that exhibit pharmacological properties as proteases inhibitors that has already been proposed as a new class of antimalarial drugs. Herein, it was tested the antimalarial property of new nine different hydroxyethylamine derivatives using the green fluorescent protein (GFP)-expressing Plasmodium berghei strain. By comparing flow cytometry and microscopic analysis to evaluate parasitaemia recrudescence, it was observed that flow cytometry was a more sensitive methodology. The nine hydroxyethylamine derivatives were obtained by inserting one of the following radical in the para position: H, 4Cl, 4-Br, 4-F, 4-CH3, 4-OCH3, 4-NO2, 4-NH2 and 3-Br. The antimalarial test showed that the compound that received the methyl group (4-CH3) inhibited 70% of parasite growth. Our results suggest that GFP-transfected P. berghei is a useful tool to study the recrudescence of novel antimalarial drugs through parasitaemia examination by flow cytometry. Furthermore, it was demonstrated that the insertion of a methyl group at the para position of the sulfonamide ring appears to be critical for the antimalarial activity of this class of compounds.

Genetic divergence among maize hybrids in cerrado-pantanal ecotone / Divergência genética entre híbridos de milho no ecótono cerrado-pantanal

In maize breeding programs can arise difficulties in relation to combination capacity studies for determination of heterotic groups, which are highly correlated with genetic divergence among the parents. The aim of this study was to estimate the genetic divergence measured for nine quantitative morphological traits in eleven single-cross hybrids of maize cultivated in the Cerrado-Pantanal ecotone. The experiment was conducted at Universidade Estadual de Mato Grosso do Sul ­ University Unit of Aquidauana. The experimental design was a randomized blocks with four replications. At harvest time, it were measured the following traits: plant height, ear insertion height, ear length, ear diameter, stem diameter, number of kernels per rows, number of rows per ear, weight of hundred grains and grain yield. In the application of hybrids cluster technique was adopted the Mahalanobis's generalized distance as dissimilarity measure, and for establishment of similar groups was applied the Tocher's method. The results indicated the existence of genetic variability among tested hybrids. The greatest genetic divergence was observed among the pairs MAXIMUS and XB6012, implying in heterotic gains. Crossings of lines extracted from hybrids 2B587HX and XB6012 with lines obtained from the other hybrids provide greater heterosis. The traits grain yield and ear insertion height were those who more and less contributed, respectively, for genetic divergence among hybrids.

Em programas de melhoramento em milho podem surgir dificuldades quanto a estudos de capacidade de combinação para determinação de grupos heretóticos, que estão altamente correlacionados com a divergência genética entre os genitores. O objetivo deste trabalho foi estimar a divergência genética mensurada para nove descritores morfológicos quantitativos em onze híbridos simples de milho cultivado no ecótono Cerrado-Pantanal. O experimento foi conduzido na área experimental da Universidade Estadual de Mato Grosso do Sul, Aquidauana (MS). O delineamento experimental utilizado foi o de blocos casualizados com quatro repetições. No momento da colheita, foram mensurados os seguintes descritores: altura de planta, altura de inserção da primeira espiga, diâmetro da espiga, diâmetro do colmo, número de grãos por fileira, número de fileiras por espiga, massa de cem grãos e rendimento de grãos. Na aplicação da técnica de agrupamento de híbridos foi adotada a distância generalizada de Mahalanobis como medida de dissimilaridade, e para o estabelecimento de grupos similares foi aplicado o método de Tocher. Os resultados indicaram a existência de variabilidade genética entre os híbridos testados. A maior divergência genética foi observada entre os pares MAXIMUS e XB6012, implicando em ganho heterótico. Cruzamentos de linhagens extraídas dos híbridos 2B587HX e XB6012 com linhagens obtidas a partir dos demais híbridos proporcionaram maior heterose. Os descritores rendimento de grãos e altura de inserção da espiga foram os que mais e menos contribuíram, respectivamente, para a divergência genética entre os híbridos.

Estudo dos mecanismos responsáveis pelo acúmulo de linfócitos durante a pleurisia induzida por Mycobacterium bovis-BCG / Study of the responsible mechanisms for the accumulation of lymphocytes during the induced pleurisy for Mycobacterium bovis-BCG

Neste trabalho nós avaliamos o mecanismo pelo qual linfócitos T acumulam na cavidade pleural de camundongos nas diferentes fases da resposta inflamatória induzida pelo BCG.Na 1ªetapa deste trabalho avaliamos o papel de moléculas de adesão da família das selectinas no acúmulo de células T na cavidade pleural após o estímulo de BCG. A injeção intratorácica(i.t.)...O pré-tratamento in vivo com fucoidina foi capaz de inibir o acúmulo de granulócitos em 24 horas e 15 dias após estímulo,mas somente inibiu o acúmulo de linfócitos em 24 horas,mas não em 15 dias.Através do modelo de transferência de células,foi observado que linfócitos marcados com FITC migram 24 horas,mas não 15 dias,após a injeção i.t.de BCG.A expressão de moléculas envolvidas na proliferação de linfócitos como a IL-2,seu receptor CD25 e o monômero que compõe o complexo AP-1 o c-Fos,foi avaliada 7 dias após a estimulação com BCG.O conteúdo de IL-2 no lavado pleural recolhido de camundongos injetados com BCG foi maior que o observado no grupo controle.A expressão de CD25 e c-Fos em linfócitos pleurais recolhidos de camundongos estimulados com BCG também foi maior que o observado no grupo estimulado com salina.O lavado pleural recolhido 7 dias após a estimulação com BCG foi capaz de induzir a proliferação de esplenócitos derivados de animais normais devido a ação da IL-2 e IFN-γ.Em conjunto,estes resultados sugerem que o acúmulo de linfócitos na cavidade pleural observada 15 dias após administração de BCG é devido a um evento proliferativo. A 2ªparte deste estudo foi dedicada a avaliar a atividade quimiotática sobre linfócitos do CCL2/CCR2 durante a pleurisia induzida por BCG.A injeção i.t.de BCG aumenta a produção de CCL2 por células pleurais em 24 horas que começa a cair em 7 dias,e retorna a níveis basais em 15 dias.Células T γσ recolhidas da cavidade pleural de camundongos estimulados com BCG expressam maiores níveis de CCR2 quando comparado com...