Increased angiogenesis parallels cardiac tissue remodelling in experimental acute Trypanosoma cruzi infection

BACKGROUND Angiogenesis has been implicated in tissue injury in several noninfectious diseases, but its role in Chagas disease (CD) physiopathology is unclear. OBJECTIVES The present study aimed to investigate the effect of Trypanosoma cruzi infection on cardiac angiogenesis during the acute phase of experimental CD. METHODS The signalling pathway involved in blood vessel formation and cardiac remodelling was evaluated in Swiss Webster mice infected with the Y strain of T. cruzi. The levels of molecules involved in the regulation of angiogenesis, such as vascular endothelial growth factor-A (VEGF-A), Flk-1, phosphorylated extracellular-signal-regulated protein kinase (pERK), hypoxia-inducible factor-1α (HIF-1α), CD31, α-smooth muscle actin (α-SMA) and also the blood vessel growth were analysed during T. cruzi infection. Hearts were analysed using conventional histopathology, immunohistochemistry and western blotting. FINDINGS In this study, our data demonstrate that T. cruzi acute infection in mice induces exacerbated angiogenesis in the heart and parallels cardiac remodelling. In comparison with noninfected controls, the cardiac tissue of T. cruzi-infected mice presented higher levels of (i) HIF-1α, VEGF-A, Flk-1 and pERK; (ii) angiogenesis; (iii) α-SMA+ cells in the tissue; and (iv) collagen -1 deposition around blood vessels and infiltrating throughout the myocardium. MAIN CONCLUSIONS We observed cardiac angiogenesis during acute experimental T. cruzi infection parallels cardiac inflammation and remodelling.

Transforming growth factor-ß as a therapeutic target for the cardiac damage of Chagas disease

Transforming growth factor beta (TGF-β) is deeply involved on the pathogenesis of Chagas disease. Our group has been investigating the participation of this pleiotropic cytokine in different aspects of Chagas disease over the last 20 years. Important observations have been made, such as: (i) the ability of Trypanosoma cruzi in activating latent TGF-β; (ii) the potential involvement of TGF-β pathway on T. cruzi invasion of host cells; (iii) association of TGF-β with parasite intracellular replication; (iv) cardiac fibrosis development and maintenance; (v) disruption of Connexin-43 plaque structures and (vi) inflammation and immune response. In this perspective article we intend to discuss the advances of the potential use of new therapies targeting TGF-β to treat the cardiac alterations of Chagas disease-affected patients.

Soil macrofauna in green manures preceding cotton growing / Macrofauna do solo em adubos verdes antecessoras ao cultivo do algodão

The objective of this work was to evaluate the abundance of the soil macrofauna in four green manure species, before the cotton crop. The experimental design was a randomized block design with four treatments: Mucuna pruriens (gray mucuna), Canavalia ensiformes (pork bean), Cajanus cajan (pigeon pea) and Crotalaria juncea (crotalaria). The macrofauna was captured by means of collections with collections using pitfall traps. The data were processed through the cluster analysis to verify the similarity among green manure species as to the occurrence of soil macrofauna. The orders Orthoptera, Coleoptera L (Larval), Hymenoptera and Coleoptera (adult), respectively, had the highest local relative abundance. There was a significant effect of the cover plants on the taxonomic groups and relative density of the soil macrofauna. Green manures were more determinant in the abundance and relative density of the soil macrofauna than the region climate. Up to 60 days after sowing the cotton crop, there was a beneficial influence of the green manures on the soil macrofauna.

Objetivou-se avaliar a abundância da macrofauna do solo em quatro espécies de adubos verdes, antecedendo a cultura do algodão. O experimento foi instalado na Universidade Estadual de Mato Grosso do Sul - Unidade Experimental de Aquidauana, MS. O delineamento experimental adotado foi em blocos ao acaso com quatro tratamentos: Mucuna pruriens (mucuna cinza), Canavalia ensiformes (feijão de porco), Cajanus cajan (guandu anão)e Crotalaria juncea (crotalária) e quatro repetições.A macrofauna foi capturada por meio de coletas com armadilhas modelo Pitfall.Os dados foram processados mediantes a análise de agrupamento para verificar a similaridade entre as espécies de adubação verde quanto à ocorrência da macrofauna do solo.A maior abundância relativa local foram as ordens, Orthoptera, Coleoptera L (Larva), Hymenoptera e Coleoptera respectivamente. Houve efeito significativo das plantas de cobertura sobre os grupos taxonômicos e densidade relativa da macrofauna edáfica. Os adubos verdes foram mais determinantes na abundância e densidade relativa da macrofauna do solo que o clima da região. Até os 60 dias de implantação da cultura do algodão, houve influência benéfica dos adubos verdes sobre a população da macrofauna do solo.

Influence of green manures on epiedaphic macrofauna in cerrado/pantanal ecotone / Influência de adubos verdes sobre a macrofauna epiedáfica no ecótono cerrado/pantanal

The aim of this study was to determine the similarity between nine species of green manure regarding the occurrence of epiedaphic macrofauna. The experiment was conducted during the months of August 2014 to July 2015 at the Crop Science sector of the State University of Mato Grosso do Sul, Unit of Aquidauana. Treatmentes evaluated were: Canavalia ensiformes, Dolichos lablab, Mucuna pruriens, Mucuna aterrima, Crotalaria spectabilis, Crotalaria juncea, Crotalaria ochroleuca, Cajanus cajan, Cajanus cajan and two control (fallow area and native vegetation). Experimental design was completely randomized with two replications. Each plot consists of 10 lines with 4.5 m long. Epiedaphic macrofauna inhabiting the interphase between the vegetation cover and the soil surface was collected weekly by the Pitfall model. Individuals collected in all green manure species in the same month were added and subjected to cluster analysis. The months of September and October composed group 2 due to higher averages of epiedaphic macrofauna individuals. The month of July had the highest number of individuals from Isoptera order, composing the group 3. The month of November was in which was collected the greatest number of individuals from Hymenoptera order, which has allocated this month in group 4. The others months of the year formed the group 1, indicating that the region climatic conditions in this period do not affect significantly the distribution of epiedaphic macrofauna individuals.

O objetivo deste trabalho foi verificar a similaridade entre nove espécies de adubo verde quanto à ocorrência de macrofauna epiedáfica. O experimento foi conduzido durante os meses de agosto de 2014 a julho de 2015 no setor de Fitotecnia da Universidade Estadual de Mato Grosso do Sul, Unidade de Aquidauana. Os tratamentos avaliados foram: Canavalia ensiformes, Dolichos lablab, Mucuna pruriens, Mucuna aterrima, Crotalaria spectabilis, Crotalaria juncea, Crotalaria ochroleuca, Cajanus cajan, Cajanus cajan e duas testemunhas (área de pousio e mata nativa). O delineamento experimental utilizado foi o inteiramente casualizado, com duas repetições. Cada parcela foi composta por 10 linhas com 4,5 m de comprimento. A macrofauna epiedáfica, que habita na interfase entre a cobertura vegetal e a superfície do solo foi coletada semanalmente por meio de armadilhas modelo Pitfall (de queda). Os indivíduos coletados em todas as espécies de adubação verde no mesmo mês foram somados e submetidos à análise de agrupamento. Os meses de setembro e outubro compuseram o grupo 2 devido as maiores médias de indivíduos da macrofauna epiedáfica. O mês de julho obteve o maior número de indivíduos da ordem Isoptera, compondo o grupo 3. O mês de novembro foi no qual foi coletado o maior número de indivíduos da ordem Hymenoptera, o que alocou este mês no grupo 4. Os demais meses do ano formaram o grupo 1, o que indica que as condições climáticas da região neste período não afetam significativamente a distribuição dos indivíduos da macrofauna epiedáfica.

Preservação da fertilidade e dos ovários em mulheres com tumores anexiais benignos / Preservation of the fertility and the ovaries in women with benign adnexal tumors

OBJETIVO: Avaliar a preservação da fertilidade e dos ovários em mulheres submetidas à cirurgia por tumor anexial benigno. MÉTODOS: Para este estudo observacional com coleta prospectiva foram incluídas 206 mulheres operadas no CAISM-Unicamp de fevereiro de 2010 a janeiro de 2014. A preservação da fertilidade foi definida como tumorectomia ou anexectomia unilateral sem histerectomia em mulheres na pré-menopausa. A preservação ovariana foi considerada quando pelo menos um ovário ou parte dele foi preservado. RESULTADOS: Das 206 mulheres com tumores anexiais benignos, 120 (58%) estavam na pré-menopausa e 86 (42%) na pós-menopausa. Na pré-menopausa, foram encontrados 36 (30%) tumores de células germinativas, 31 (26%) neoplasias epiteliais e 11 (9%) do cordão sexual e estroma. Na pós-menopausa foram identificados 35 (41%) neoplasias epiteliais, 27 (31%) do cordão sexual e estroma e 8 (9%) de células germinativas. Entre as 36 mulheres com tumores ovarianos não neoplásicos, 21 (58%) apresentavam endometriomas e 8 (22%) cistos funcionais. Das 22 mulheres com tumores extra ovarianos, o leiomioma uterino foi o achado mais frequente (50%). Entre as pacientes com ≤35 anos, 26 (57%) foram submetidas à tumorectomia e 18 (39%) a anexectomia unilateral com preservação do útero e anexo contralateral. Mulheres com ≤35 anos foram mais frequentemente operadas por laparoscopia que esteve associada a maior taxa de preservação de fertilidade quando comparada com a laparotomia (p<0,01). Observou-se que 26 das pacientes submetidas à histerectomia com anexectomia (28%) bilateral estavam na pré-menopausa. CONCLUSÕES: Embora se observe uma tendência em realizar apenas tumorectomia em mulheres com ≤35 anos, uma proporção significativa de mulheres jovens ainda é ...

PURPOSE: To evaluate the sparing of fertility and ovaries in women submitted to surgical treatment for benign adnexal tumors. METHODS: Between February 2010 and January 2014, 206 patients were included in this observational study as they were submitted to surgical treatment for benign ovarian tumors at CAISM, a tertiary hospital. Fertility sparing surgery was defined as tumorectomy or unilateral salpingoophorectomy without hysterectomy in premenopausal women. Preservation of the ovary occurred when at least one ovary or part of it was mantained. RESULTS: Of the 206 women with benign tumors, 120 (58%) were premenopausal and 86 (42%) were postmenopausal. There were 36 (30%) ovarian germ cell tumors, 31 (26%) epithelial neoplasms and 11 (9%) sex-cord stromal tumors among premenopausal women. In the group of postmenopausal women, 35 (41%) epithelial neoplasms, 27 (31%) sex-cord stromal tumors and 8 (9%) ovarian germ cell tumors were identified. Among 36 women with non-neoplastic ovarian tumors, 21 (58%) had endometriomas and 8 (22%) functional cysts. Among 22 women with extra-ovarian tumors, uterine leiomyomatosis was the most frequent finding (50%). In the group of women who were ≤35 years old, 26 (57%) were treated by tumorectomy and 18 (39%) were submitted to unilateral salpingoophorectomy with sparing of the uterus and the contralateral ovary. Women who were ≤35 years old were more frequently operated by laparoscopy which was associated with a higher number of fertility sparing procedures when compared to laparotomy (p<0.01). Twenty-six (28%) women submitted to hysterectomy with bilateral salpingoophorectomy were premenopausal. CONCLUSION: Although there is a trend to perform only tumorectomy in women who are ≤35 years old, a significant number of young women is still treated by salpingoophorectomy. Among 36- to 45-year-old women, only 70% had their fertility spared, while 20% had both ovaries removed. ...

Experimental chemotherapy for Chagas disease: 15 years of research contributions from in vivo and in vitro studies

Chagas disease, which is caused by the intracellular parasite Trypanosoma cruzi, is a neglected illness with 12-14 million reported cases in endemic geographic regions of Latin America. While the disease still represents an important public health problem in these affected areas, the available therapy, which was introduced more than four decades ago, is far from ideal due to its substantial toxicity, its limited effects on different parasite stocks, and its poor activity during the chronic phase of the disease. For the past 15 years, our group, in collaboration with research groups focused on medicinal chemistry, has been working on experimental chemotherapies for Chagas disease, investigating the biological activity, toxicity, selectivity and cellular targets of different classes of compounds on T. cruzi. In this report, we present an overview of these in vitro and in vivo studies, focusing on the most promising classes of compounds with the aim of contributing to the current knowledge of the treatment of Chagas disease and aiding in the development of a new arsenal of candidates with anti-T. cruzi efficacy.

Morte celular & diamidinas: estudos em Trypanossoma Cruzi e Leishmania L. Amazonensis / Cellular death & diamidines: studies in Trypanossoma Cruzi and Leishmania L. Amazonensis

O conceito de morte abrange múltiplas interpretações sob diferentes aspectos incluindo filosófico, orgânico e legal, sendo definido em termos médicos como término das funções vitais: “parada definitiva e irreversível dos processos metabólicos, das funções orgânicas e das atividades vitais”. A biologia celular utiliza-se de diferentes ferramentas bioquímicas, ópticas e ultra-estruturais, entre outras, para identificar células nas quais as funções vitais deixaram de existir. O conhecimento gerado nestes estudos permitiu identificar que este processo não implica necessariamente na morte do indivíduo, pois diariamente milhões de células morrem e são substituídas sem que haja maiores implicações. A morte celular ocorre em (i) processos fisiológicos, como parte importante no desenvolvimento embrionário e fetal, na renovação das células e em vários aspectos da maturação dos mecanismos imunológicos de defesa e, em (ii) processos patológicos, como infecções parasíticas, nas quais o surgimento de alterações estruturais passa a comprometer a integridade do órgão e a manutenção de suas funções. Com relação a patogenias causadas por diversos agentes (incluindo bactérias, fungos e protozoários), sabe-se que a integridade e a forma de morte das células do hospedeiro e do próprio parasita representam fatores determinantes para o desenvolvimento e manutenção da doença. Sob este aspecto, a presente tese abordará dois blocos: o primeiro avaliando morte celular envolvida na infecção pelo T. cruzi, agente etiológico de uma importante parasitose endêmica em países da América Latina, e o segundo bloco analisando a morte celular induzida pela ação de diamidinas aromáticas que são compostos ativos no tratamento de diversas infecções parasíticas, incluindo o Trypanosoma cruzi e L.(L) amazonensis, dois parasitas de grande relevância médica, cujas patologias ainda incuráveis atingem cerca de 30 milhões de indivíduos.

In vitro measurement of enzymatic markers as a tool to detect mouse cardiomyocytes injury

Primary cultures of cardiomyocytes represent a useful model for analyzing cardiac cell biology as well as pathogenesis of several cardiovascular disorders. Our aim was to standardize protocols for determining the damage of cardiac cells cultured in vitro by measuring the creatine kinase and its cardiac isotype and lactate dehydrogenase activities in the supernatants of mice cardiomyocytes submitted to different protocols of cell lysis. Our data showed that due to its higher specificity, the cardiac isotype creatine kinase was the most sensitive as compared to the others studied enzymatic markers, and can be used to monitor and evaluate cardiac damage in in vitro assays.