Abstract Chagas disease is a neglected parasitic disease caused by Trypanosoma cruzi, whose treatment has remained unsatisfactory for over 50 years, given that it is limited to two drugs. Benznidazole (BZN) is an efficient antichagasic drug used as the first choice, although its poor water-solubility, irregular oral absorption, low efficacy in the chronic phase, and various associated adverse effects are limiting factors for treatment. Incorporating drugs with such characteristics into nanostructured lipid carriers (NLC) is a promising alternative to overcome these limiting obstacles, enhancing drug efficacy and bioavailability while reducing toxicity. Therefore, this study proposed NLC-BZN formulations in different compositions prepared by hot-melt homogenization followed by ultrasound, and the optimized formulation was characterized by FTIR, DRX, DSC, and thermogravimetry. Biological activities included in vitro membrane toxicity (red blood cells), fibroblast cell cytotoxicity, and trypanocidal activity against epimastigotes of the Colombian strain of T. cruzi. The optimized NLC-BZN had a small size (110 nm), negative zeta potential (-18.0 mV), and high encapsulation (1.64% of drug loading), as shown by infrared spectroscopy, X-ray diffraction, and thermal analysis. The NLC-BZN also promoted lower in vitro membrane toxicity (<3% hemolysis), and 50% cytotoxic concentration (CC50) for NLC-BZN in L929 fibroblast cells (110.7 µg/mL) was twice the value as the free BZN (51.3 µg/mL). Our findings showed that the NLC-BZN had higher trypanocidal activity than free BZN against the epimastigotes of the resistant Colombian strain, and this novel NLC-BZN formulation proved to be a promising tool in treating Chagas disease and considered suitable for oral and parenteral administration. cruzi 5 years (BZN choice watersolubility, watersolubility water solubility, solubility water-solubility absorption phase NLC (NLC obstacles Therefore NLCBZN hotmelt hot melt ultrasound FTIR DRX DSC thermogravimetry red cells, , cells) cytotoxicity T 110 (11 nm, nm nm) 18.0 180 18 0 (-18. mV, mV mV) 1.64% 164 1 64 (1.64 loading, loading loading) spectroscopy Xray X ray diffraction analysis <3% 3 (<3 hemolysis, hemolysis hemolysis) CC50 CC (CC50 L L92 110.7 1107 7 (110. µg/mL µgmL µg mL 51.3 513 51 (51. µg/mL. . administration 11 (1 18. (-18 1.64 16 6 (1.6 <3 (< CC5 (CC5 L9 110. 51. (51 ( (-1 1.6 (1. < (CC (5 (- 1.

Resumo Senna rugosa (Fabaceae) é uma espécie comum do território brasileiro, especialmente no Cerrado. É amplamente utilizado na medicina popular, embora ainda não tenha sido investigada quanto aos seus fitoconstituintes e atividades farmacológicas. Neste estudo, foram realizadas as análises fitoquímicas preliminares do extrato etanólico de folhas de S. rugosa e sua citotoxicidade, atividade antitumoral e antimicrobiana (antibacteriana, antifúngica, anti-Trypanosoma e anti-Leishmania). O extrato bruto foi analisado em HPLC-DAD e as frações foram caracterizadas usando técnicas de GC-MS e ESI-MS. Os resultados indicam a presença majoritária de compostos fenólicos, incluindo flavonóis, antraquinonas e derivados de antronas. Atividades antimicrobianas foram detectadas contra as espécies Staphylococcus aureus, Pseudomonas aeruginosa, Candida famata, Candida krusei e Candida tropicalis. Além disso, uma fraca atividade anti-tripanossomatídeos contra as cepas M2269 e BH46 de Leishmania sp. Uma atividade antitumoral foi identificada, porém também foi observada uma toxicidade sobre células de fibroblastos (L929). Fitoquimicamente, os resultados fornecem evidências de que compostos fenólicos nas folhas de S. rugosa podem ser os responsáveis pela sua atividade antimicrobiana e pela toxicidade celular observada em nosso estudo.

Abstract Senna rugosa (Fabaceae) is a common specie of the Brazilian territory, especially in the Cerrado biome. It is widely used in the popular medicine although not yet adequately investigated as to its phytoconstituents and pharmacological activities. In this study, the preliminary phytochemical analysis of ethanolic leaf extract of S. rugosa was performed and its cytotoxicity, antitumoral and antimicrobial activities (antibacterial, antifungal, anti-Trypanosoma and anti-Leishmania) were evaluated. The crude extract was analyzed in HPLC-DAD and fractions were also characterized using GC-MS and ESI-MS techniques. Results indicate phenolic compounds majority presence, including flavonol, anthraquinone and anthrones derivatives. Relative antimicrobial activities were detected against Staphylococcus aureus, Pseudomonas aeruginosa, Candida famata, Candida krusei and Candida tropicalis. As well as, a weak anti-trypanosomatids activity against M2269 strain and BH46 strain of Leishmania sp. A relative antitumoral activity was also identified but in counterpoint were also observed a toxicity in fibroblast cells (L929). Phytochemically, results provide evidence that phenolic compounds in S. rugosa leaves might be the responsible for its antimicrobial activity and cell toxicity inferred in our research.

RESUMO Objetivo: O presente estudo teve como objetivo avaliar a presença de helmintos e protozoários intestinais em hortaliças comercializados em Diamantina, um município localizado no Vale do Jequitinhonha, uma das regiões mais pobres do mundo. Métodos: Cento e oito exemplares, incluindo alface, cebolinha e rúcula, foram mensalmente coletados em uma feira livre, uma quitanda e um supermercado do município. As amostras foram processadas por um método de concentração e avaliadas por microscopia óptica para pesquisa de estruturas parasitárias. Resultados: O percentual global de contaminação foi de 50,9% (55/108), com predominância de larvas de nematódeos (36,5%), cistos de Entamoeba coli e ovos de ancilostomídeos/Strongyloides spp. (12,9%). A alface demonstrou a maior taxa de contaminação (61,1%) e as amostras da feira livre foram as mais contaminadas (77,8%). Informações coletadas em cada ponto de venda apontaram o cultivo em campo como a etapa crítica para a contaminação. Conclusão: Hortaliças comercializadas em Diamantina apresentam uma ampla variedade de parasitas intestinais, o que representa um risco potencial à saúde dos consumidores da área.

ABSTRACT Objective: The present study aimed to evaluate the presence of helminthes and intestinal protozoa in vegetables commercialized in Diamantina, a municipality located at Jequitinhonha Valley, one of the poorest regions of the world. Methods: A total of 108 specimens, including lettuce, green onion and rocket, were monthly collected from the most popular open street market, green grocery and supermarket of the municipality. The samples were processed by a concentration method and evaluated by light microscopy for parasitological identification. Results: The percentage of contamination was 50.9% (55/108), with predominance of nematode larvae (36.5%), cysts of Entamoeba coli (26.0%) and eggs of hookworms/Strongyloides spp. (12.9%). Lettuce showed greater contamination rate (61.1%) and samples from the open street market were more contaminated (77.8%). Information collected at each point of sale pointed the field cultivation as the critical step for such contaminations. Conclusion: Vegetables marketed in Diamantina presents a wide variety of intestinal parasites, which may represent a potential risk to the health of consumers of fresh vegetables.

Reports of triatomine infestation in urban areas have increased. We analysed the spatial distribution of infestation by triatomines in the urban area of Diamantina, in the state of Minas Gerais, Brazil. Triatomines were obtained by community-based entomological surveillance. Spatial patterns of infestation were analysed by Ripley’s K function and Kernel density estimator. Normalised difference vegetation index (NDVI) and land cover derived from satellite imagery were compared between infested and uninfested areas. A total of 140 adults of four species were captured (100 Triatoma vitticeps, 25Panstrongylus geniculatus, 8 Panstrongylus megistus, and 7 Triatoma arthurneivai specimens). In total, 87.9% were captured within domiciles. Infection by trypanosomes was observed in 19.6% of 107 examined insects. The spatial distributions ofT. vitticeps, P. geniculatus, T. arthurneivai, and trypanosome-positive triatomines were clustered, occurring mainly in peripheral areas. NDVI values were statistically higher in areas infested by T. vitticeps and P. geniculatus. Buildings infested by these species were located closer to open fields, whereas infestations of P. megistus andT. arthurneivai were closer to bare soil. Human occupation and modification of natural areas may be involved in triatomine invasion, exposing the population to these vectors.

Trypanosoma cruzi strains from distinct geographic areas show differences in drug resistance and association between parasites genetic and treatment response has been observed. Considering that benznidazole (BZ) can reduce the parasite burden and tissues damage, even in not cured animals and individuals, the goal is to assess the drug response to BZ of T. cruzi II strains isolated from children of the Jequitinhonha Valley, state of Minas Gerais, Brazil, before treatment. Mice infected and treated with BZ in both phases of infection were compared with the untreated and evaluated by fresh blood examination, haemoculture, polymerase chain reaction, conventional (ELISA) and non-conventional (FC-ALTA) serologies. In mice treated in the acute phase, a significant decrease in parasitaemia was observed for all strains. Positive parasitological and/or serological tests in animals treated during the acute and chronic (95.1-100%) phases showed that most of the strains were BZ resistant. However, beneficial effect was demonstrated because significant reduction (p < 0.05%) and/or suppression of parasitaemia was observed in mice infected with all strains (acute phase), associated to reduction/elimination of inflammation and fibrosis for two/eight strains. BZ offered some benefit, even in not cured animals, what suggest that BZ use may be recommended at least for recent chronic infection of the studied region.

Introduction The biological diversity of Trypanosoma cruzi strains plays an important role in the clinical and epidemiological features of Chagas disease. Methods Eight T. cruzi strains isolated from children living in a Chagas disease vector-controlled area of Jequitinhonha Valley, State of Minas Gerais, Brazil, were genetically and biologically characterized. Results The characterizations demonstrated that all of the strains belonged to T. cruzi II, and showed high infectivity and a variable mean maximum peak of parasitemia. Six strains displayed low parasitemia, and two displayed moderate parasitemia. Later peaks of parasitemia and a predominance of intermediate and large trypomastigotes in all T. cruzi strains were observed. The mean pre-patent period was relatively short (4.2±0.25 to 13.7±3.08 days), whereas the patent period ranged from 3.3±1.08 to 34.5±3.52 days. Mortality was observed only in animals infected with strain 806 (62.5%). Histopathological analysis of the heart showed that strains 501 and 806 caused inflammation, but fibrosis was observed only in animals infected with strain 806. Conclusions The results indicate the presence of an association between the biological behavior in mice and the genetic characteristics of the parasites. The study also confirmed general data from Brazil where T. cruzi II lineage is the most prevalent in the domiciliary cycle and generally has low virulence, with some strains capable of inducing inflammatory processes and fibrosis.

INTRODUÇÃO: O objetivo foi desenvolver um método sorológico in-house de alta especificidade e sensibilidade para diagnosticar e monitorar a morbidade da doença de Chagas. MÉTODOS: Para tal, a reatividade sorológica de IgG e subclasses foi testada em soros de pacientes chagásicos de Berilo, Vale do Jequitinhonha/MG/Brasil. A reatividade sorológica foi também avaliada em amostras de pacientes com outras doenças infecto-contagiosas relevantes, incluindo o HIV, vírus da hepatite C (VHC), sífilis (SYP), leishmaniose visceral (LV), leishmaniose tegumentar americana (LTA) e controles não infectados (NI) para verificar o desempenho do método. Outras análises foram feitas para avaliar a aplicabilidade desta metodologia no monitoramento da morbidade da doença de Chagas. Com este propósito os pacientes com doença de Chagas foram anteriormente classificados em três grupos: indeterminados (IND), cardíacos (CARD) e digestivos/mistos (DIG/Mis) conforme seu estado clínico. RESULTADOS: A análise da reatividade sorológica de IgG total na diluição 1:40 mostrou ser uma abordagem importante no diagnóstico da doença de Chagas (100% de sensibilidade e especificidade e ausência de reação cruzada com as demais infecções). A análise das subclasses de IgG mostrou reação cruzada principalmente com NI, LV e LTA em todas as diluições. O grupo IND apresentou a maior reatividade para IgG3 e o grupo DIG/Mis apresentou nível mais elevado de IgG se comparados aos grupos IND e CARD. CONCLUSÕES: Estes achados demonstram que o método de ELISA in-house apresenta uma promissora aplicabilidade no diagnóstico diferencial e na avaliação da morbidade da doença de Chagas.

INTRODUCTION: The goal was to develop an in-house serological method with high specificity and sensitivity for diagnosis and monitoring of Chagas disease morbidity. METHODS: With this purpose, the reactivities of anti-T. cruzi IgG and subclasses were tested in successive serum dilutions of patients from Berilo municipality, Jequitinhonha Valley, Minas Gerais, Brazil. The performance of the in-house ELISA was also evaluated in samples from other relevant infectious diseases, including HIV, hepatitis C (HCV), syphilis (SYP), visceral leishmaniasis (VL), and American tegumentary leishmaniasis (ATL), and noninfected controls (NI). Further analysis was performed to evaluate the applicability of this in-house methodology for monitoring Chagas disease morbidity into three groups of patients: indeterminate (IND), cardiac (CARD), and digestive/mixed (DIG/Mix), based on their clinical status. RESULTS: The analysis of total IgG reactivity at serum dilution 1:40 was an excellent approach to Chagas disease diagnosis (100% sensitivity and specificity). The analysis of IgG subclasses showed cross-reactivity, mainly with NI, VL, and ATL, at all selected serum dilutions. Based on the data analysis, the IND group displayed higher IgG3 levels and the DIG/Mix group presented higher levels of total IgG as compared with the IND and CARD groups. CONCLUSIONS: These findings demonstrated that methodology presents promising applicability in the analysis of anti-T. cruzi IgG reactivity for the differential diagnosis and evaluation of Chagas disease morbidity.