RESUMO: O tumor venéreo transmissível canino (TVTC) é uma neoplasia transmissível de elevada casuística no Brasil. A quimioterapia com sulfato de vincristina é considerada o tratamento de escolha, mas a necessidade de aplicações semanais e acompanhamento hematológico, além dos custos, são obstáculos à adesão dos proprietários ao tratamento. A lomustina é um antineoplásico da classe dos agentes alquilantes e, por ser administrado por via oral, representa um opção de tratamento mais prática e menos onerosa para os proprietários de animais com neoplasias. O objetivo deste estudo foi avaliar a eficácia terapêutica da lomustina em cães acometidos por TVTC. Foram selecionados 12 cães com diagnóstico citopatológico de TVTC genital de ocorrência natural. Os cães foram submetidos ao protocolo experimental com administração de lomustina nas doses de 70 a 85 mg/m2 por via oral a cada 21 dias, totalizando no máximo dois ciclos de administração. Os animais foram reavaliados a cada sete dias até um máximo de +49 dias após a primeira dose de lomustina, para monitorar a regressão das lesões neoplásicas por meio de mensuração das lesões. Entre os 12 cães submetidos ao protocolo, 8/12 obtiveram remissão completa da neoplasia e foram considerados curados (66,6%), 1/12 tiveram resposta parcial ao tratamento (8,33%) e 3/12 tiveram doença estável (25%). Efeitos adversos importantes, como leucopenia neutrofílica grave, foram detectados em 3/12 cães (25%). O estudo clínico indicou que a lomustina pode ser uma opção de tratamento para TVTC.

ABSTRACT: Canine transmissible venereal tumor (TVTC) is a highly casuistic transmissible neoplasm in Brazil. Chemotherapy with vincristine sulfate is considered the treatment of choice, but the need for weekly applications and hematological monitoring, in addition to costs, are obstacles to owners’ adhesion to the treatment. Lomustine is an alkylating class antineoplastic agent, and because it is administered orally, it is a more practical and less costly treatment option for the owners of animals with neoplasms sensitive to the drug. This study evaluated the therapeutic efficacy of lomustine in dogs affected by TVTC. Twelve dogs with cytopathological diagnosis of natural genital TVTC were selected. The dogs were submitted to the experimental protocol with lomustine administration at doses of 70 to 85 mg/m2 orally every 21 days, totaling a maximum of two administration cycles. The animals were reevaluated every 7 days until a maximum of +49 days after the first dose of lomustine, to monitor the regression of neoplastic lesions through measurements. Among the 12 dogs submitted to the lomustine protocol, 8/12 achieved complete remission of the neoplasm and were considered cured (66.6%), 1/12 had partial response to treatment (8.33%) and 3/12 had stable disease (25%). Important adverse effects such as severe neutrophilic leukopenia were detected in 3/12 dogs (25%). The clinical study indicated that lomustine may be a treatment option for TVTC.

Resumen Introducción: La cuantificación de SARS-CoV-2 en aguas residuales es una herramienta que permite determinar la tendencia de la circulación viral en un área geográfica determinada. Objetivo: Cuantificar el virus SARS-CoV-2 en 15 plantas de tratamiento de aguas residuales en diferentes ciudades de Chile para establecer una comparación con las variables de: i) casos activos por cada 100.000 habs.; ii) positividad diaria (casos nuevos); y iii) fases del plan de confinamiento. Metodología: SARS-CoV-2 se concentró a partir de muestras de aguas residuales. Para obtener el número de genomas del virus por litro se realizó una cuantificación absoluta utilizando qRT-PCR. Resultados: Entre enero y junio de 2021 se procesaron 253 muestras, siendo todas positivas para la presencia del virus. Asimismo, se logró determinar que la tasa de casos activos por cada 100.000 habs. es la variable que mejor se ajusta a las tendencias obtenidas con la cuantificación de la carga viral en las aguas residuales. Conclusiones: La cuantificación de SARS-CoV-2 en las aguas residuales de manera permanente es una herramienta eficiente para determinar la tendencia del virus en un área geográfica determinada y, en conjunto con una vigilancia genómica, puede constituirse en una vigilancia centinela ideal generando alertas sobre futuros brotes.

Abstract Background: The quantification of SARS-CoV-2 in wastewater is a tool that allows determining the trend of viral circulation in a particular geographical area. Aim: To quantify the SARS-CoV-2 virus in 15 wastewater treatment plants in different Chilean cities to establish a comparison with the variables of: i) Active cases per 100,000 inhabitants; ii) daily positivity (novel cases); and iii) phases of the lockdown strategy. Methods: SARS-CoV-2 was concentrated from wastewater samples. To obtain the number of virus genomes per liter, absolute quantification was performed using qRT-PCR. Results: Between January and June 2021, 253 samples were processed, all of which were positive for the presence of the virus. Likewise, it will be determined that the rate of active cases per 100,000 inhabitants is the variable that best fits the trends obtained with the quantification of the viral load in wastewater. Conclusions: The quantification of SARS-CoV-2 in wastewater as a continuous strategy is an efficient tool to determine the trend of the viral circulation in a delimited geographical area and, combined with genomic surveillance, it can constitute an ideal sentinel surveillance alert on future outbreaks.

Resumo Intervenções para a redução do estigma relacionado às pessoas que usam drogas podem melhorar sua qualidade de vida e adesão ao tratamento. Esta revisão teve por objetivo identificar e analisar estudos de intervenções para reduzir o estigma relacionado às pessoas que usam drogas. Foram realizadas buscas nas bases PubMed, APA PsycNET, ScienceDirect, Web of Science e BVS, sem delimitação de ano. Foram identificados 5.488 registros somados à busca secundária de referências. Esta revisão abrange 28 artigos e foi baseada no PRISMA. Houve variação no tamanho da amostra e intervenções, embora o público-alvo de estudantes e profissionais da saúde e o uso de métodos quantitativos tenham sido predominantes. Limitações metodológicas consideráveis foram identificadas nos estudos. Portanto, conclui-se que não há evidências que permitam inferir que as intervenções propostas são efetivas e quais delas deveriam continuar a ser empregadas para essa finalidade. É imprescindível investir em abordagens distintas das adotadas tradicionalmente.

Abstract Interventions to reduce stigma related to people who use drugs can improve their quality of life and adherence to treatment. This review aimed to identify and analyze studies on interventions to reduce the stigma related to people who use drugs. Searches were performed in the following databases, without period delimitation: PubmMed, APA PsycNET, ScienceDirect, Web of Science, and VHL. Considering secondary referencing, we identified a total of 5,488 records. This review covered 28 articles and was based on PRISMA. Although target audiences comprising students and health professionals were predominant, as well as the use of quantitative methods, sample size and interventions varied among studies. The selected studies present considerable methodological limitations. We concluded that no evidence confirms the effectiveness of the proposed interventions and which of them should continue to be applied for this specific aim. It is essential to invest in approaches other than those traditionally adopted.

Resumen Las intervenciones para la reducción del estigma relacionado a las personas que usan drogas pueden mejorar su calidad de vida y la adherencia al tratamiento. Esta revisión tuvo como objetivo identificar y analizar los estudios sobre las intervenciones para la reducción del estigma relacionado a las personas que usan drogas. Se realizaron búsquedas en las bases de datos PubMed, APA PsycNET, ScienceDirect, Web of Science y BVS, sin delimitación del año. Se identificaron 5.488 registros sumados a la búsqueda secundaria de referencias. Esta revisión abarca 28 artículos y se basó en PRISMA. Hubo una variación en el tamaño de la muestra y las intervenciones, aunque fueron predominantes el público objetivo de estudiantes y profesionales de la salud y el uso de métodos cuantitativos. Se identificaron limitaciones metodológicas considerables en los estudios. Se concluye que no hay evidencias de que las intervenciones propuestas son efectivas y cuáles deberán seguir siendo utilizadas para este propósito. Es esencial invertir en enfoques distintos de los adoptados tradicionalmente.

RESUMO Este estudo teve como objetivo caracterizar a cadeia produtiva do leite e estudar a sanidade do rebanho leiteiro do município de Codó, estado do Maranhão, Brasil, através da aplicação de um checklist e da realização de exames para detecção de mastite clínica e subclínica pelos métodos da caneca do fundo escuro e California mastitis test (CMT), no período de junho a agosto de 2019, em 295 vacas leiteiras procedentes de 20 propriedades. Coletaram-se amostras de leite e água da ordenha para análises microbiológicas. Observou-se que os ordenhadores não possuem conhecimento adequado sobre as boas práticas agropecuárias. Quanto à realização da ordenha, apenas 60% realizam-na em currais com cobertura; quanto ao piso, 60% são de terra batida e 40% de concreto. A água utilizada no manejo da ordenha é proveniente de poços em 35% das propriedades e as demais de açudes. As práticas de pré e pós-dipping e os testes CMT e caneca de fundo escuro e a adoção de programa de controle da mastite não são realizados em nenhuma das propriedades. Diagnosticaram-se duas vacas com mastite subclínica e uma com tuberculose e verificou-se elevada contaminação por coliformes totais e termotolerantes nas análises microbiológicas do leite, variando entre 23 NMP/mL a >1.100 NMP/mL e < 3.0 a > 1.100 NMP/mL, respectivamente, e presença de estafilococos coagulase positivos em 25% das amostras. As amostras de água também apresentaram elevada contaminação por coliformes totais entre 4,1 NMP/mL a > 2.419,6 NMP/mL e 40% apresentaram presença de Escherichia coli. Esses resultados refletem a necessidade de mais investimentos em assistência técnica e treinamento técnico para esses produtores.

ABSTRACT In order to characterize the milk production chain and study the dairy herd health in the city of Codó, state of Maranhão, Brazil, a checklist was applied and tests were carried out to detect clinical and subclinical mastitis using mastitis test strip cup and the California mastitis test (CMT), from June to August 2019, in 295 dairy cows from 20 farms. Water and milk samples were collected for microbiological analysis. It was observed that herdsmen do not have adequate knowledge about good agricultural practices. As for milking, only 60% are performed in corrals with coverage, and as for the floor, 60% are made of clay and 40% are made of concrete. In 35% of the properties, the water used in milking management comes from wells and the rest from dams. Pre and postdipping practices, CMT, mastitis strip cup test and the adoption of a mastitis control program are not carried out on any of the properties Two cows tested positive for subclinical mastitis and one cow tested positive for tuberculosis. In the microbiological analysis of the milk, a high count of total coliforms and thermotolerants was obtained, with values between 23 to > 1,100 MPN/mL and < 3.0 to > 1,100 MPN/mL, respectively. The presence of coagulase positive staphylococci was also observed in 25% of the samples. The water samples also showed high contamination by total coliforms between 4.1 to > 2.419.6 MPN/mL and 40% showed the presence of Escherichia coli. These results reflect the need for more investments in technical assistance and technical training for these producers.

ABSTRACT Ultraviolet B radiation has been described as a potential elicitor agent of the synthesis of secondary metabolites in plants. Thus, the present study aimed to investigate the production of betalains and total flavonoids, as well as the antioxidant activity, of Alternanthera sessilis, A. brasiliana, A. tenella and A. philoxeroides exposed to different periods of UV-B radiation (280-315 nm). Plants of these four species were exposed to UV-B radiation for 0, 2, 4, 6, and 8 hours, which amounts to 0, 10, 20, 30, and 40 J cm−2 of radiation, respectively. Significant increases in betacyanin and betaxanthin levels were observed in A. sessilis and A. brasiliana during the period of UV-B exposure, while no differences were observed for the others species. The highest estimated flavonoid levels were for A. sessilis exposed to UV-B radiation for 8 h, followed by a 24-h recovery period. In conclusion, the action of UV-B radiation as an inducer of defence responses in plants is influenced by increasing exposure periods followed by a recovery period. Both increase the levels of these composts, yet this increase is different among the four Alternanthera species, having a greater influence on the species A. sessilis and A. brasiliana.

Abstract Trifolium pratense L., Fabaceae, is a rich source of isoflavones and has become the focus of several studies related to its phytoestrogenic activity. The aim of this study was to establish germination and cell cultures protocol for T. pratense and quantify isoflavones content in cell cultures, in vitro cultured and wild plants harvested in two different seasons. Murashige Skoog medium supplemented with naphthalene acetic acid and kinetin was able to produce the highest formation of friable calli. Calli cultures were analyzed qualitatively after 60 days of culture, and in vitro plants after 30, 45 and 60 days of cultivation. The chemical analysis was performed by ultra performance liquid chromatography, using the linearity curves of daidzein, genistein, formononetin and biochanin A as standards. The concentrations of isoflavones detected in wild plants were different in the two harvest periods and contrasted in content when compared to the in vitro plants. Cell cultures exhibited diverse profiles and concentration of isoflavones, none of which presented the isoflavonoid biochanin A. Pectinase was used to promote reduction of clumps and ended up altering the characteristics of secondary metabolites production in some cultures. Formononetin showed higher concentration in wild red clover samples (15.407 mg g-1), and in the in vitro grown plants the highest concentration was daidzein (17.591 mg g-1) at 60 days. The methods used for this research were effective, and the red clover plants of the analyzed variety can be cultivated in vitro aiming the commercial productivity by having contents greater than or equal to the wild plants in the periods studied, even without the use of elicitors during the cultivation.

RESUMO O objetivo deste estudo foi estabelecer um protocolo para indução de calos e produção de betacianinas em plantas de Alternanthera brasiliana. Para indução da calogênese foram utilizados explantes de A. brasiliana e cinco combinações de citocinina e auxina. Posteriormente os calos foram transferidos para Meio de Indução de Betacianinas (MIB), formado por meio MS, com 0,5 mg L-1 de tidiazuron (TDZ) e 1 mg L-1 de ácido naftalenoacético (ANA), onde permaneceram na luz por 45 dias. Avaliou-se oaspecto e a intensidade de pigmentos e quantificação de betacianinas totais em Cromatografia Líquida de Alta Eficiência (CLAE). Os segmentos internodais combinados com o meio que contém equilíbrio na concentração de auxinas e citocininas foi mais eficiente para obtenção de calogênese e maior produção de betacianinas. A presença de amarantina, em calos de A. brasiliana, justifica seu uso medicinal e contribui para futuros estudos para produção em larga escala desta molécula.

ABSTRACT The objective of this study was to establish a protocol for callus induction and betacyanin production in plants of Alternanthera brasiliana. Explants of A. brasiliana and five combinations of cytokinin and auxin were used for callus induction. Calli were transferred to a Betacyanin Induction Medium (MIB), composed of MS, with 0.5 mg L-1 of thidiazuron (TDZ) and 1 mg L-1 of naphthaleneacetic acid (NAA), and kept in the light for 45 days. The aspect and intensity of pigments were assessed and total betacyanins were quantified in high performance liquid chromatography (HPLC). The combination of internodal segments and a medium containing equilibrium concentrations of auxins and cytokinins was the most efficient metod to induce calli and increased production of betacyanins. The presence of amaranthine in calli of A. brasiliana justifies its medical use and the consequent need for future studies for the large-scale production of this molecule.

Resumo O objetivo deste trabalho foi avaliar a expressão gênica de citocinas pró-inflamatórias (RANKL, TNF-a e IFN-g) e regulatórias (TGF-b e IL-10) em resposta à infecção experimental por Fusobacterium nucleatum (ATCC 10953) e Enterococcus faecalis (ATCC 19433) como mono-infecção ou em bi-associação. F. nucleatum e E. faecalis foram inoculados no sistema de canais radiculares de camundongos Balb/c, tanto isoladas como em bi-associação. Os animais foram sacrificados em 10 e 20 dias após a infecção, e os tecidos periapicais foram coletados. As expressões do mRNA das citocinas RANKL, TNF-a, IFN-g, TGF-b e IL-10 foram analisadas por meio do real-time PCR. O teste de Kruskal-Wallis foi utilizado para análise estatística. A mono-infecção com F. nucleatum induziu alta expressão de RANKL e TNF-a, enquanto sua modulação ocorreu devido à IL-10. A alta expressão de IFN-g no dia 20 foi regulada positivamente por E. faecalis e RANKL; TNF-a foi regulada positivamente por um mecanismo independente via IL-10 e TGF-b. A bi-associação (F. nucleatum e E. faecalis) estimulou uma alta expressão de RANKL, TNF-a e IFN-g, que parece ser modulada por TGF-b após 20 dias. A expressão gênica de citocinas pró-inflamatórias foi mais proeminente nos estágios iniciais da infecção periapical experimental, com concomitante redução no período tardio. Este fenômeno pode ser regulado por IL-10 e TGF-b em uma condição de infecção específica.

Abstract The aim of this study was to evaluate the gene expression of proinflammatory (RANKL, TNF-a and IFN-g) and regulatory (TGF-b and IL-10) cytokines as reaction to experimental infection by mono or bi-association of Fusobacterium nucleatum (ATCC 10953) and Enterococcus faecalis (ATCC 19433). F. nucleatum and E. faecalis, either in mono- or bi-association were inoculated into the root canal system (RCS) of Balb/c mice. Animals were sacrificed at 10 and 20 days after infection and periapical tissues surrounding the root were collected. The mRNA expression of the cytokines RANKL, TNF-a, IFN- g, TGF-b and IL-10 was assessed using real-time PCR. The Kruskal-Wallis test was used for statistical analysis. F. nucleatum mono-infection induced high expression of RANKL and TNF-a, while its modulation was due to IL-10. High expression of IFN-g at day 20 was up-regulated by E. faecalis and RANKL; TNF-a was up-regulated by an independent mechanism via IL-10 and TGF-b. Bi-association (F. nucleatum and E. faecalis) stimulated high expression of RANKL, TNF-a and IFN-g, which seemed to be modulated by TGF-b 20 days later. The gene expression of proinflammatory cytokines was more prominent in the earlier periods of the experimental periapical infection, which concomitantly decreased in the later period. This expression may be regulated by IL-10 and TGF-b in an infection-specific condition