Experimental Chemotherapy for Chagas Disease: A Morphological, Biochemical, and Proteomic Overview of Potential Trypanosoma cruzi Targets of Amidines Derivatives and Naphthoquinones

Chagas disease (CD), caused by Trypanosoma cruzi, affects approximately eight million individuals in Latin America and is emerging in nonendemic areas due to the globalisation of immigration and nonvectorial transmission routes. Although CD represents an important public health problem, resulting in high morbidity and considerable mortality rates, few investments have been allocated towards developing novel anti-T. cruzi agents. The available therapy for CD is based on two nitro derivatives (benznidazole (Bz) and nifurtimox (Nf)) developed more than four decades ago. Both are far from ideal due to substantial secondary side effects, limited efficacy against different parasite isolates, long-term therapy, and their well-known poor activity in the late chronic phase. These drawbacks justify the urgent need to identify better drugs to treat chagasic patients. Although several classes of natural and synthetic compounds have been reported to act in vitro and in vivo on T. cruzi, since the introduction of Bz and Nf, only a few drugs, such as allopurinol and a few sterol inhibitors, have moved to clinical trials. This reflects, at least in part, the absence of well-established universal protocols to screen and compare drug activity. In addition, a large number of in vitro studies have been conducted using only epimastigotes and trypomastigotes instead of evaluating compounds' activities against intracellular amastigotes, which are the reproductive forms in the vertebrate host and are thus an important determinant in the selection and identification of effective compounds for further in vivo analysis. In addition, due to pharmacokinetics and absorption, distribution, metabolism, and excretion characteristics, several compounds that were promising in vitro have not been as effective as Nf or Bz in animal models of T. cruzi infection. In the last two decades, our team has collaborated with different medicinal chemistry groups to develop preclinical studies for CD and investigate the in vitro and in vivo efficacy, toxicity, selectivity, and parasite targets of different classes of natural and synthetic compounds. Some of these results will be briefly presented, focusing primarily on diamidines and related compounds and naphthoquinone derivatives that showed the most promising efficacy against T. cruzi

CULTURA DE SEGURIDAD ENTRE PROFESIONALES DE SALUD EN HOSPITAL DE ENSEÑANZA

Objetivo: Evaluar la cultura de seguridad organizacional entre los profesionales de un hospital de enseñanza. Método: Estudio tipo survey con 645 profesionales de un hospital de enseñanza en Estado de Paraná, de octubre de 2014 a julio de 2015, con instrumento auto aplicable de Agency for Health care Research and Quality. Variables cuantitativas fueron representadas en porcentajes; cuando ≥75% de las respuestas fueron positivas, cultura de seguridad satisfactoria; 74 a 51% neutra; y ≤50% débil. La confiabilidad fue mensurada por el coeficiente Alfa de Cronbach. Resultado: Mayor índice fue obtenido en la dimensión “Expectativas sobre el supervisor/jefe y acciones promotoras da seguridad”, con 70,8% (n=455) de respuestas positivas y “Respuestas no punitivas al error” el menor, con 25,6% (n=164). El coeficiente Alfa de Cronbach obtuvo media de 0,62, presentando baja confiabilidad. Conclusión: Los resultados muestran cultura de seguridad insatisfactoria y relevancia de acciones de promoción, destacándose abordaje colectivo y no punitivo de errores.Objective: assess the safety culture among health professionals of a teaching hospital. Method: Cross-sectional surveytype study with 645 professionals of a teaching hospital of the State of Paraná, from October 2014 to July 2015, through the use of a self-administered instrument of the Agency for Health care Research and Quality. Quantitative variables were represented in percentage values: the safety culture was considered satisfactory if ≥75% of the responses were positive; neutral, if 74 to 51% of the responses were positive, and fragile, if ≤50% of the responses were positive. Reliability was measured by Cronbach’s alpha coefficient. Results: the highest value was obtained for dimension “Expectations about your supervisor/boss and safety-promoting actions”, with 70.8% (n=455) of positive responses and “Non punitive responses to error” obtained the lowest value, with 25.6% (n=164). The Cronbach’s alpha coefficient obtained an average of 0.62, showing low reliability Conclusion: The results reveal an unsatisfactory safety culture and stress the importance of actions targeted to the promotion of a safety culture, with emphasis to a collective and non-punitive approach to errors.Objetivo: avaliar a cultura de segurança organizacional entre os profissionais de um hospital de Ensino. Método: estudo tipo survey com 645 profissionais de um hospital de ensino no Estado do Paraná, de outubro de 2014 a julho de 2015, utilizando instrumento autoaplicável da Agency for Health care Research and Quality. Variáveis quantitativas foram representadas em percentuais; quando ≥75% das respostas foram positivas, cultura de segurança satisfatória; 74 a 51% neutra; e ≤50% frágil. A confiabilidade foi mensurada pelo coeficiente Alfa de Cronbach. Resultados: maior índice foi obtido na dimensão “Expectativas sobre o seu supervisor/chefe e ações promotoras da segurança”, com 70,8% (n=455) de respostas positivas e “Respostas não punitivas ao erro” o menor, com 25,6% (n=164). O coeficiente Alfa de Cronbach obteve média de 0,62, mostrando baixaconfiabilidade. Conclusão: resultados mostram cultura de segurança insatisfatória e relevância de ações de promoção, com destaque à abordagem coletiva e não punitiva de erros

True Prevalence and Spatial Distribution of Equine Infectious Anemia Virus (EIAV) in Horses from Northeast Region of Brazil

Background: Equine infectious anemia (EIA) is a viral infection, caused by a lentivirus of the Retroviridae family, Orthoretrovirinael subfamily and its occurrence generates significant economic losses due to culling of positive animals as a measure of infection control. The objective of this work was to determine the prevalence of horses positive for equine infectious anemia virus (EIAV) and to identify the occurrence of areas with higher densities of cases in the states of Paraíba (PB), Pernambuco (PE), Rio Grande do Norte (RN) and Ceará (CE), Northeast region of Brazil, during the rainy (May and June) and dry (October and November) periods of 2017 and 2018.Materials, Methods & Results: Serum samples from 6,566 horses from the states of PB, PE, RN and CE, Brazil, provided by the Laboratório Veterinária Diagnóstico - Ltda., were used. Serological diagnosis of EIA was performed using indirect enzyme-linked immunosorbent assay (ELISA) as a screening test and agar gel immunodiffusion test (AGID) as a confirmatory test. The apparent prevalence was obtained by dividing the number of seroreactive animals by the total number of animals, while the true prevalence was estimated by adjusting the apparent prevalence, considering the sensitivity (100%) and specificity (98.6%) of the diagnostic protocol used. For the construction of Kernel estimates, the Quartic function was used. In the dry season, of the 1,564 animals sampled, 28 were serologically positive, of which 19 belonged to the state of Ceará, 7 to Paraíba and 2 to Rio Grande do Norte. In 2018, it was observed that, during the rainy season, 26 of the 1,635 horses were seroreactive, with 19 cases resulting from Ceará, 4 from Paraíba and 3 from Pernambuco. In the dry season, 32 of the 1,526 animals were seroreactive to EIAV, of which 26 were from Ceará, 3 from Paraíba, 1 from Rio Grande do Norte and 2 from Pernambuco. In the dry period of 2017, the CE had a real prevalence of 1.22% (95% CI = 0.05 - 2.99%). In 2018, during the rainy season, prevalences of 0.03% (95% CI = 0 - 1.18%) were identified in CE and 1.69% (95% CI = 0 - 8.38%) in PE. Regarding the 2018 dry period, a prevalence of 1.32% (95% CI = 0.26 - 2.84%) was found in the state of CE. In both dry and rainy periods of 2017, the presence of spatial clusters of animals positive for EIA was observed, mainly in the border areas among the states of CE, PE, PB and RN. In 2018, there was a variation in the distribution of areas with higher densities of cases between the rainy and dry periods.Discussion: The state of CE had the highest prevalence of positive animals and the presence of areas with higher densities of EIA cases in both climatic periods, in the years 2017 and 2018. In some municipalities of the CE, important sporting events of agglomeration of animals take place, which can favor the transmission of EIAV by facilitating the contact of infected and susceptible animals. Population density may be a factor associated with the higher prevalence observed in this region, as it has the second largest herd among the states studied. Higher densities indirectly contribute to the occurrence of infectious diseases, as they favor the contact of infected and susceptible animals. The occurrence of higher densities of cases in the border areas of the states of PE, RN, CE, and PB may be related to the greater movement of animals in these regions, favoring the indirect contact of infected horses with susceptible ones. The observed results demonstrate the circulation of the EIAV in four states in the Northeast region of Brazil.Keywords: Equine infectious anemia virus, prevalence, spatial clusters, epidemiology

FENOLOGIA DE Vellozia ramosissima L. B. Sm. (VELLOZIACEAE) EM ÁREAS DE "COMPLEXO RUPESTRE" QUARTZÍTICO E FERRUGINOSO

O estudo teve como objetivo compreender a fenologia reprodutiva e vegetativa da espécie Vellozia ramosissima, associando o padrão fenológico  s variações sazonais caracterí­sticas do ambiente. O estudo foi realizado em áreas do complexo rupestre quartzí­tico e ferruginoso no municí­pio de Conceição do Mato Dentro, MG. Foram observadas mensalmente as caracterí­sticas fenológicas de vinte plantas, no perí­odo de jul/2015 a jul/2016. Foram utilizados dados meteorológicos de precipitação, temperatura, umidade e velocidade do vento para o perí­odo de estudo. O padrão fenológico foi determinado pelo método de intensidade de Fournier, correlação de Spearman e análise circular. Foram encontradas correlações significativas entre as fenofases e as variáveis abióticas apenas para dispersão dos frutos e produção de folhas novas. A espécie apresentou um comportamento fenológico bastante sazonal para maior parte dos eventos, reflexo da sazonalidade climática, caracterí­stica da região de estudo. Foram observados padrões distintos entre as áreas para todas as fenofases avaliadas

GERMINAÇÃO DE SEMENTES DE Vellozia ramosissima L. B. Sm. (VELLOZIACEAE) EM RESPOSTA íâ?¬ DIFERENTES TEMPERATURAS

Devido   escassez de estudos acerca do gênero Vellozia e sua importância ecológica nos Complexos Rupestres da Cadeia do Espinhaço, este trabalho teve como objetivo avaliar a germinação das sementes de canela-de-ema (Vellozia ramosissima L.B.Sm.), submetidas  s temperaturas constantes de, 20í°C, 30í°C, 40í°C e 50í°C.   O delineamento experimental adotado foi inteiramente casualizado, com quatro repetições, sendo cinquenta amostras por repetição. A contagem da germinação foi realizada durante 30 dias. Os parâmetros avaliados foram a porcentagem, o í­ndice de velocidade e o tempo médio de germinação.   A germinação foi nula a 50í°C, e a faixa de germinabilidade ocorreu nas temperaturas de 20, 30 e 40í°C. A ocorrência de germinação da espécie em uma maior amplitude de temperatura sugere a capacidade de colonização numa maior diversidade de hábitats, indicando adaptação da mesma a ambientes expostos ao sol e   substratos sombreados, como fendas de rochas

O sistema nacional de inspeção e vigilância sanitária para a agricultura familiar e empresarial no Brasil

Neste trabalho é analisado o Sistema Nacional de Inspeção e Vigilância Sanitária - SNIVS, por meio do conjunto de normas que o constitui, assim como, de sua operacionalização enquanto instrumentos de regulação das atividades de produção, distribuição e consumo de alimentos de origem agrícola e pecuária no Brasil. O objetivo desta pesquisa é compreender o funcionamento do Sistema Nacional de Inspeção e de Vigilância Sanitária–SNIVS, suas vinculações institucionais e sua atuação na agricultura familiar e empresarial no Brasil, no contexto da discussão da concentração fundiária. O método utilizado foi à análise documental. Nos resultados da pesquisa identificam-se dois períodos distintos que caracterizam a atuação dos órgãos e entidades que compõem o SNIVS: num primeiro momento, suas atuações são favoráveis a Agricultura Empresarial, num segundo momento, as ações daqueles órgãos e entidades favorecem aos interesses da Agricultura Familiar. A pesquisa mostrou também que o SNIVS está operando um cuidadoso processo de harmonização institucional e operacional de acordo com os tratados bilaterais de comércio firmados com a Organização Mundial do Comércio – OMC e com a Organização Mundial da Saúde – OMS

Validation of an analytical method by high-performance liquid chromatography and microbiological assay, biological safety and in silico toxicity for danofloxacin

Danofloxacin is a veterinary fluoroquinolone used to treat respiratory and gastrointestinal diseases of birds, pigs and cattle. The literature reviewed shows some analytical methods to quantify this fluoroquinolone, but microbiological and biological safety studies are limited. The analytical methods were validated by the Official Codes. The LC-DAD method was developed and validated using an RP-18 column, mobile phase containing a mixture of 0.3% triethylamine (pH 3.0) and acetonitrile (85:15, v/v). The microbiological assay was performed by agar diffusion method (3 x 3) and Staphylococcus epidermidis as a microorganism test. Forced degradation studies were performed in both methods. The minimum inhibitory concentration (MIC) was performed by test microdilution and toxicity studies were evaluated using in silico study, cell proliferation, cell viability test, micronuclei and comet assay. LC and a microbiological assay proved linear, accurate, precise, and robust to quantify danofloxacin, but only the LC method showed selectivity to quantify the drug in the presence of its degradation products. These results demonstrate that the LC method is suitable for stability studies of danofloxacin, but a microbiological assay cannot be used to quantify the drug due to the biological activity of the photoproducts. Ex-vivo cytotoxicity and theoretical and experimental genotoxicity were also observed

The soil-plant interface in Megathyrsus maximus cv. Mombasa subjected to different doses of nitrogen in rotational grazing

Este trabajo tuvo como objetivo evaluar los efectos de tres dosis de nitrógeno (N) en las características morfogénicas y estructurales, la masa (MR) y la distribución de las raíces en el perfil del suelo, y la resistencia del suelo a la penetración de las pasturas de guinea Mombasa manejados con pastoreo rotacional. El diseño experimental utilizó bloques al azar con tres dosis de N (100, 200 y 300 kg ha-1) y tres repeticiones. El criterio para interrumpir el rebrote de los pastos fue la altura de 80 a 90 cm del dosel (90-95 % de intercepción de luz por el dosel). Los animales fueron retirados de los potreros cuando el dosel alcanzó el 50 % de la altura prepastoreo. Se evaluó la masa y acumulación del forraje, las características morfogénicas y estructurales del dosel, la MR y distribución en el perfil del suelo, y la resistencia del suelo a la penetración. En pastos fertilizados con 200 y 300 kg ha-1 de N, se observó la mayor aparición foliar (0.090 hojas macollo-1 día-1 y 0.081 hojas macollo-1 día-1), tasas de alargamiento (2.82 y 2.61 cm macollo-1 d-1) y mayor acumulación diaria de forraje (113.8 y 106.6 kg ha-1d-1). El uso de 300 kg ha-1 de N promovió una mayor resistencia del suelo a la penetración a 10 cm de profundidad en el pospastoreo (3.3 MPa). No se observó ningún efecto de las dosis de N en la MR (P>0.05). Por lo tanto, el control de la altura antes y después del pastoreo de los animales en los potreros ayudó a mantener la estructura del pasto y evitar el proceso de compactación del suelo.  De acuerdo con los resultados, se concluye que 200 y 300 kg ha-1 de fertilización con N es una estrategia para intensificar los pastos.This work aimed to evaluate the effects of three nitrogen (N) doses on the morphogenic and structural characteristics, root mass (RM) and distribution in the soil profile, and penetration soil resistance of Mombasa guineagrass pastures managed with rotational stocking. The experimental design used randomized blocks with three N doses (100, 200, and 300 kg ha-1) and three replicates. The criterion for interrupting the pastures’ regrowth was the height of 80 to 90 cm of the canopy (90–95 % of light interception by the canopy). The animals were removed from the paddocks when the canopy reached 50 % of the pre-grazing height. Forage mass and accumulation, canopy morphogenic and structural characteristics, RM and distribution in the soil profile, and soil resistance to penetration were evaluated. In pastures fertilized with 200 and 300 kg ha-1 of N, the highest leaf appearance  (0.090 and 0.081 leaves-1 tiller-1 d-1),  elongation rates (2.82 and 2.61 cm tiller-1 d-1) and highest daily forage accumulation (113.8 and 106.6 kg ha-1d-1) were observed. Using 300 kg ha-1 of N promoted greater soil resistance to penetration at 10 cm of depth in the post-grazing (3.3 MPa). No effect of N doses was observed for RM (P>0.05). The pre- and post-grazing height control of animals in the paddocks therefore helped to maintain the pasture structure and avoid the soil compaction process. According to the results, it is concluded that 200 and 300 kg ha-1 of N fertilization is a strategy for intensifying pastures

Por entre as várzeas da Amazônia: educação ambiental como instrumento de gestão no parque estadual charapucu/ Marajó-PA/ Among the floodplains of the Amazon: environmental education as a management tool in charapucu state park/ Marajó-PA

Este trabalho correlaciona a metodologia da gestão em Unidade de Conservação (UC) e a função da Educação Ambiental (EA) na área do Parque Estadual Charapucu (PEC), sendo uma Unidade de Conservação de Natureza de Proteção Integral localizada na Amazônia Paraense na Ilha do Marajó, situada na cidade de Afuá. O objetivo do projeto foi desenvolver ações e práticas de conscientização que visam a Educação Ambiental de maneira ampliada e singular às características das sociedades ribeirinhas marajoaras do PEC. Metodologicamente essa pesquisa apresenta abordagem qualitativa por envolver estudos de ações sociais grupais, tratando-se de uma pesquisa-ação participativa baseada nos estudos epistemológicos de LEFF (2001). O recorte temporal para o levantamento dos dados obtidos situou-se sobre um projeto ainda vigente de 12 meses no ano de 2018. A metodologia ativa parte do princípio da EA de planejamento de atividades que partem da realidade das comunidades tradicionais da região, por meio de uma abordagem interligada a UC, no sentido de tornar as populações bem mais conscientes sobre os possíveis danos ao meio ambiente. Estre trabalho traz todas as ações desenvolvidas neste período, possibilitou alcançar 15 escolas na zona rural e urbana e 12 comunidades rurais. Com a realização do referido projeto de educação ambiental as comunidades e a gestão estadual tiveram uma melhor amplitude e fortalecimento sendo possível construir um cenário direcionado à conscientização à educação ambiental

In Vitro and In Vivo Investigation of the Efficacy of Arylimidamide DB1831 and Its Mesylated Salt Form - DB1965 - against Trypanosoma cruzi Infection

Chagas disease is caused by infection with the intracellular protozoan parasite Trypanosoma cruzi. At present, nifurtimox and benznidazole, both compounds developed empirically over four decades ago, represent the chemotherapeutic arsenal for treating this highly neglected disease. However, both drugs present variable efficacy depending on the geographical area and the occurrence of natural resistance, and are poorly effective against the later chronic stage. As a part of a search for new therapeutic opportunities to treat chagasic patients, pre-clinical studies were performed to characterize the activity of a novel arylimidamide (AIA - DB1831 (hydrochloride salt) and DB1965 (mesylate salt)) against T.cruzi. These AIAs displayed a high trypanocidal effect in vitro against both relevant forms in mammalian hosts, exhibiting a high selectivity index and a very high efficacy (IC50 value/48 h of 5–40 nM) against intracellular parasites. DB1965 shows high activity in vivo in acute experimental models (mouse) of T.cruzi, showing a similar effect to benznidazole (Bz) when compared under a scheme of 10 daily consecutive doses with 12.5 mg/kg. Although no parasitological cure was observed after treating with 20 daily consecutive doses, a combined dosage of DB1965 (5 mg/kg) with Bz (50 mg/kg) resulted in parasitaemia clearance and 100% animal survival. In summary, our present data confirmed that aryimidamides represent promising new chemical entities against T.cruzi in therapeutic schemes using the AIA alone or in combination with other drugs, like benznidazole