Manipulação gênica do protozoário Neospora caninum como ferramenta para o estudo das interações parasito-hospedeiro

Neospora caninum is an obligate intracellular parasite classified in the phylum Apicomplexa, characterized by the presence of the apical complex composed by micronemes proteins, rhoptries and dense granules, used by parasite during the adhesion and invasion process of the host cell. This is the mean event in infection pathogenesis generated by N. caninum and other parasites from the phylum Apicomplexa, promoting influence in the parasite biology and the interface between the parasite and its host. Therefore, molecular tools have been developed in order to identify and characterize these possible virulence factors. Thus, the present study sought to establish a specific system of genetic manipulation of N. caninum, searching for the improvement of the genetics manipulation of this parasite. So, we developed genetically depleted N. caninum to Rop9 rhoptry using the pU6-Universal CRISPR-Cas9 plasmid of T. gondii modified by the insertion of Ku80. The Rop9 depleted parasite showed important during initial phase of invasion and replication of the parasite, however it was not characterized as a potential virulence fator for N. caninum. Furthermore, T. gondii proteins were expressed in N. caninum by the use of specific vectors for this parasite, showing an heterologous system for the study of Toxoplasma proteins, due to the fact that Gra15 or Gra24 of type II T. gondii and Rop16 of type I T. gondii were expressed in N. caninum tachyzoites in a stable way and keept its biological phenotype, as already presented the former parasite, that naturaly expresses these proteins. In addition, it was observed that N. caninum induced an inflammasome activation through NLRP3, ASC and Caspase-1. IL-1R/MyD88 demonstrated an indirect pathway in the control of parasite replication. Furthermore, it was observed that this activation is dependent of the potassium efflux and that different strains of N. caninum keep this activation profile. However, T. gondii strains block this activation, making necessary a prior signal in order to active the inflamosome pathway. Type I T. gondii Rop16 was identified as responsible for blocking this activation, in a dependent way to the STAT3 activation. Therefore, the development of molecular tools and their application in N. caninum may prove to be useful to identify and characterize virulent factors involved in the pathogenesis by these two protozoans.Conselho Nacional de Desenvolvimento Científico e TecnológicoCoordenação de Aperfeiçoamento de Pessoal de Nível SuperiorFundação de Amparo a Pesquisa do Estado de Minas GeraisTese (Doutorado)Neospora caninum é um parasito intracelular obrigatório pertencente ao filo Apicomplexa, caracterizado pela presença do complexo apical composto por proteínas de micronemas, roptrias e grânulos densos, utilizadas pelo parasito durante o processo de adesão e invasão da célula hospedeira, caracterizando o principal evento na patogênese da infecção por N. caninum e por outros parasitos do filo Apicomplexa, influenciando assim na biologia do parasito e na interação parasito-hospedeiro. Dessa forma, ferramentas moleculares tem sido desenvolvidas na tentativa de se identificar e caracterizar esses possíveis fatores de virulência. Portanto, o presente trabalho buscou desenvolver ferramentas moleculares especificas para N. caninum de modo a melhorar a eficiência na manipulação genética deste parasito, sendo assim desenvolvemos N. caninum depletado geneticamente para a roptria Rop9 por meio da utilização do plasmídeo CRISPR-Cas9 pU6-Universal de T. gondii modificado pela inserção de Ku80. O parasito depletado para Rop9 mostrou-se importante durante a fase inicial da invasão e replicação do parasito, entretanto não foi caracterizado como um potencial fator de virulência para N. caninum. Complementarmente, proteínas de T. gondii foram expressas em N. caninum utilizando vetores específicos para este parasito, apresentando um sistema heterólogo para estudo de proteínas presentes em Toxoplasma. Pois, Gra15 ou Gra24 de T. gondii tipo II, e Rop16 de T. gondii tipo I foram expressos em taquizoítos de N. caninum de forma estável e mantiveram o seu fenótipo biológico como já apresentado pelo parasito de origem, que expressa naturalmente estas proteínas. Adicionalmente, foi observado que N. caninum induziu a ativação do inflamassoma através da via NLRP3, ASC e Caspase-1. Sendo a via IL-1R/ MyD88 uma via indireta importante no controle da replicação do parasito. Além disso, foi demonstrado que essa ativação é dependente de efluxo de potássio, e que diferentes cepas de N. caninum mantêm esse perfil de ativação. No entanto, cepas de T. gondii bloqueiam essa ativação sendo necessário um primeiro sinal para acionar a via do inflamassoma. Foi identificado Rop16 de T. gondii da cepa tipo I como responsável por bloquear essa ativação, de modo dependente a ativação de STAT3. Portanto, as ferramentas moleculares desenvolvidas e aplicadas em N. caninum revelaram-se úteis para identificar e caracterizar fatores de virulência envolvidos na patogênese por estes dois protozoários

Effects and prevalence of responders after a multicomponent intervention on cardiometabolic risk factors in children and adolescents with overweight/obesity: action for health study

This study aimed to verify the effect of a multicomponent intervention on cardiometabolic risk factors (CMRF), and to determine the prevalence of responders on CMRF among children and adolescents with overweight/ obesity. This is a quasi-experimental study, developed with 35 children and adolescents with overweight/ obesity (control group (CG) = 18; intervention group (IG) = 17), aged between 7 and 13 years. Participants in IG underwent a multicomponent intervention for 12 weeks. The following variables were evaluated: anthropometric measures, maturational stages and CMRF (body fatness, HOMA-IR, triglycerides, high-density and low-density lipoprotein) (HDL-C, LDL-C), total cholesterol (TC), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and AST/ALT ratio. Mixed analysis of variance and the prevalence of responders were used for statistical analysis. There was a significant time x group interaction on body fatness (p < 0.001), HOMA-IR (p = 0.01), HDL-C (p < 0.001), LDL-C (p = 0.009) and TC (p < 0.001). The prevalence of responders for CMRF in IG and CG was respectively: body fatness (47%; 0%; p = 0.04), HOMA-IR (58.8%; 16.6%; p = 0.04); triglycerides (17.6%; 5.5%; p = 0.31); HDL-C (76.4%; 5.5%; p = 0.01), LDL-C (35.3%; 5%; p = 0.08), TC (64.7%; 5%; p = 0.01), AST (5.8%; 0%; p = 0.87), ALT (29.4%; 11.1%; p = 0.24) and AST/ALT ratio (24.4%; 22.2%; p = 0.67). Multicomponent intervention induced positive changes on CMRF along with a higher prevalence of positive adaptations in IG than the CG in some of the cardiometabolic outcomes assessed

Multicomponent intervention efect on cardiometabolic risk factors among overweight/obese Brazilian children: a mediation analysis

Purpose To verify whether percentage of body fat, physical ftness, physical activity and calorie intake mediates the multicomponent intervention efect on cardiometabolic risk factors in overweight/obese children, and present the relative contribution of each mediator. Methods This is an intervention study, developed with 35 overweight/obese school-aged children (control group=17 and intervention group=18), aged between 7 and 13 years (9.05±1.90). A 12-week multicomponent intervention was performed, consisting of physical exercise, nutritional education sessions and parental support. The following variables were evaluated at baseline and post-intervention: anthropometric measures and percentage of body fat, physical ftness, physical activity assessed by accelerometer, total calorie intake and biochemical assays. For statistical analysis, generalized linear models were used. Results The intervention efect on glucose was mediated by percentage of body fat (24%), muscular ftness (22%) and total calorie intake (40%). The same was observed for alanine aminotransferase (ALT), with a mediation proportion of 26, 31 and 35%, respectively, as well as for HDL-C (percentage of body fat −30%, muscular ftness −30% and total calorie intake −33%); while vigorous physical activity mediated the intervention efect on glucose (40%), HDL-C (39%) LDL-C (43%) and total cholesterol (37%). Conclusion Interventions strategies should focus on reducing percentage of body fat and calorie intake, and enhancing muscular ftness and vigorous physical activity to achieve efective changes on cardiometabolic risk factors

24- hour movement behaviors and fitness in preschoolers: acompositional and isotemporal reallocation analysis

This study analyzed the associations between the 24-hour movement behaviors composition and fitness in preschoolers and investigated predicted changes in fitness when time in active behaviors is reallocated. This cross-sectional study was carried out with 270 preschoolers (132 boys; 3.97 ± 0.80 years-old). Light and moderate-to-vigorous physical activity (LPA and MVPA), and sedentary behavior (SB) were verified using an accelerometer. Sleep time was obtained through interviews with parents. Components of physical fitness (cardiorespiratory fitness—CRF, speed-agility, and lower-body muscular strength) were assessed using the PREFIT Battery. To verify the association between 24-hour movement behaviors and physical fitness, the compositional analysis was used, and for the time reallocation, the compositional isotemporal substitution analysis was used for active behaviors (LPA and MVPA). The daily composition, adjusted for body mass index, sex, and age, was significantly associated with CRF (P = .007; r2 = 0.29), speed-agility (P < .001; r2 = 0.14), and lower-body muscular strength (P = .01; r2 = 0.07). For CRF, the addition of MVPA, at the expense of any other behavior, was associated with significant improvements. For speed-agility and lower-body muscular strength, only reallocations between sleep and LPA yielded significant associations. The variation in CRF, speed-agility, and lower-body muscular strength was associated with the 24 hours movement composition, and reallocating 5, 10 or 15 minutes of SB or sleep for MVPA was significantly positive for CRF (P < .05). The present findings highlight the relevance of decreasing SB and increasing physical activity practice, particularly at high intensities, to promote a better CRF profile for preschoolers

Fitoterápicos com pontencial de ação antiparasitária presentes na baixada maranhense / Phytotherapeutics with antiparasitic action potential present in the maranhense lowlands

O Brasil contempla uma vasta biodiversidade vegetal, sendo classificado como um ponto tangencial em potencial no desenvolvimento de novas substâncias terapêuticas de origem natural. Este estudo evidencia uma revisão da literatura norteada pelo objetivo de esclarecer a utilidade terapêutica de cinco plantas pertencentes à Baixada Maranhense em relação a infecções parasitárias tanto no homem quanto em animais de interesse econômico, que são a arruda, abóbora, mangueira, o mastruço e cajueiro, a qual advém da compilação de dados resultantes de artigos científicos, trabalhos de conclusão de curso, além de teses de mestrado e doutorado. Foi constatado que essas plantas denotam de potencial ação antiparasitária, mas os trabalhos que investigam essa atividade ainda são incipientes, e em sua maioria consistem em ensaios pré-clínicos ou análises de modelos experimentais. Desse modo, mais pesquisas devem acontecer para que o potencial desses agentes fitorerápicos frente a parasitoses possa ser comprovada devidamente, e os estudos já publicados precisam continuar, dando prosseguimento aos ensaios clínicos

Sueño y salud mental infantil: papel de la actividad física y aptitud cardiorrespiratoria

Introduction: Sleep is considered an important health indicator and plays a key role in brain development and plasticity. Objective: To ascertain whether there is a relationship between sleep quality and mental health indicators and whether organized physical activity (PA) or cardiorespiratory fitness (CRF) act as moderators of this association. Methods: This is a cross-sectional study with a quantitative approach. The sample consisted of 226 students between six and 11 years of age, male and female, in the early years of elementary education at a public school in Porto Alegre, Brazil, which was selected for convenience. CRF was measured by running test and six-minute walk. Sleep quality and organized physical activity outside of school were verified through an anamnesis, socioeconomic status through an adaptation of the ABEP (Brazilian Association of Research Companies) questionnaire, and mental health indicators with the assistance of the Strengths and Difficulties Questionnaire, all answered by parents. Frequencies, means, standard deviations and generalized linear models were used for the data analysis, while a 95% confidence interval was used for the analyses. Results: Sleep quality was associated with total difficulties (β=7.659, p<0.001), emotional symptoms (β= 1.754; p=0.001), hyperactivity/inattentiveness (β=3.054, p <0.001), conduct problems (β=1.619, p<0.001) and peer relationship problems (β=1.231; p=0.007) in boys. In girls it was related to total difficulties (β=3.421; p=0.006), and conduct problems (β=1.235; p=0.003). However, the interactions were not significant. Conclusion: Although they occur independently, stress is placed on the importance of sleeping well, engaging in organized PA and having good CRF levels for the improvement and maintenance of mental health.O sono é considerado um importante indicador da saúde e tem papel fundamental no desenvolvimento e na plasticidade cerebral. Verificar se há relação entre a qualidade do sono e os indicadores de saúde mental e se a atividade física (AF) organizada ou aptidão cardiorrespiratória (APCR) atuam como moderadores dessa associação. Trata-se de um estudo de corte transversal com abordagem quantitativa. A amostra foi composta por 226 alunos entre seis a 11 anos de idade do sexo feminino e masculino, estudantes dos anos iniciais do ensino fundamental de uma escola pública de Porto Alegre - Brasil, sendo a mesma selecionada por critério de conveniência. A APCR foi mensurada através do teste de corrida e da caminhada de seis minutos. A qualidade do sono e a AF organizada fora da escola foram verificadas através de uma anamnese, o nível socioeconômico através de uma adaptação do questionário da ABEP (Associação Brasileira de Empresas de Pesquisa) e os indicadores da saúde mental com o auxílio do Strengths and Difficulties Questionnaire, todos respondidos pelos pais. Para a análise de dados, utilizaram-se frequências, médias e desvios padrão e modelos lineares generalizados. Para as análises, utilizou-se um intervalo de confiança de 95%. Resultados: A qualidade do sono apresentou associação com o total de dificuldades (β=7,659; p<0,001), sintomas emocionais (β=1,754; p=0,001), hiperatividade/déficit de atenção (β=3,054; p<0,001), problemas de conduta (β=1,619; p<0,001) e problemas de relacionamento com colegas (β=1,231; p=0,007), nos meninos. Já nas meninas, apresentou relação com o total de dificuldades (β=3,421; p=0,006) e problemas de conduta (β=1,235; p=0,003). No entanto, as interações não foram significativas. Conclusão: Apesar de ocorrerem por vias independentes, destaca-se a importância de dormir bem, praticar AF organizada e ter bons níveis de APCR para a melhora e manutenção da saúde mental.Introducción: El sueño es considerado un indicador importante de la salud y juega un papel fundamental en el desarrollo y la plasticidad cerebral. Objetivo: Verificar si hay relación entre la calidad del sueño y los indicadores de salud mental y si la actividad física (AF) organizada o aptitud cardiorrespiratoria (APCR) actúan como moderadores de esa asociación. Métodos: Se trata de un estudio de corte transversal, con abordaje cuantitativo. La muestra fue compuesta por 226 alumnos entre seis a 11 años de edad del sexo femenino y masculino, estudiantes de los años iniciales de la enseñanza fundamental de una escuela pública de Porto Alegre - Brasil, siendo la misma seleccionada por criterio de conveniencia. La APCR se midió a través del test de carrera y de caminata de seis minutos. La calidad del sueño y la AF organizada fuera de la escuela fueron verificadas a través de una anamnesis, el nivel socioeconómico a través de una adaptación del cuestionario de la ABEP (sigla para Asociación Brasileña de Empresas de Encuestas), y los indicadores de salud mental con la ayuda de Strengths and Difficulties Questionnaire, todos respondidos por los padres. Para el análisis de datos, se utilizaron frecuencias, promedios y desviaciones estándar y modelos lineales generalizados. Para los análisis, se utilizó un intervalo de confianza de 95%. Resultados: La calidad del sueño presentó asociación con el total de dificultades (β = 7,659, p <0,001), síntomas emocionales (β = 1,754; p = 0,001), hiperactividad/déficit de atención (β = 3,054; p<0,001) problemas de conducta (β=1,619, p<0,001), y problemas de relacionamiento con colegas (β = 1,231; p = 0,007) en los niños. En las niñas, presentó relación con el total de dificultades (β = 3,421; p = 0,006) y problemas de conducta (β = 1,235; p = 0,003). Sin embargo, las interacciones no fueron significativas. Conclusión: A pesar de ocurrir por vías independientes, se destaca la importancia de dormir bien, practicar AF organizada y tener buenos niveles de APCR para la mejora y mantenimiento de la salud mental

TNF-TNFR1 signaling enhances the protection against Neospora caninum infection

Neospora caninum is a protozoan associated with abortions in ruminants and neuromuscular disease in dogs. Classically, the immune response against apicomplexan parasites is characterized by the production of proinflammatory cytokines, such as IL-12, IFN-γ and TNF. TNF is mainly produced during the acute phases of the infections and binds to TNF receptor 1 (CD120a, p55, TNFR1) activating a variety of cells, hence playing an important role in the induction of the inflammatory process against diverse pathogens. Thus, in this study, we aimed to evaluate the role of TNF in cellular and humoral immune responses during N. caninum infection. For this purpose, we used a mouse model of infection based on wildtype (WT) and genetically deficient C57BL/6 mice in TNFR1 (Tnfr1-/-). We observed that Tnfr1-/- mice presented higher mortality associated with inflammatory lesions and increased parasite burden in the brain after the infection with N. caninum tachyzoites. Moreover, Tnfr1-/- mice showed a reduction in nitric oxide (NO) levels in vivo. We also observed that Tnfr1-/- mice showed enhanced serum concentration of antigen-specific IgG2 subclass, while IgG1 production was significantly reduced compared to WT mice, suggesting that TNFR1 is required for regular IgG subclass production and antigen recognition. Based on our results, we conclude that the TNF-TNFR1 complex is crucial for mediating host resistance during the infection by N. caninum

Acetonic Fraction of Bidens pilosa Enriched for Maturase K Is Able to Control Cerebral Parasite Burden in Mice Experimentally Infected With Toxoplasma gondii

Toxoplasma gondii infection can cause abortions or congenital infection for a vast number of domestic animals and humans, leading to economic loss in veterinary sciences, as well as severe consequences for immunocompromised patients. Bidens pilosa Linné has been used in ethnopharmacology for treatment of diseases, as malaria, diabetes and hepatitis, in addition to its use as antioxidant, antiallergic, anti-inflammatory, and antiviral. The components of this plant have never been studied before for treatment of toxoplasmosis, and the conventional drugs currently used to treat this disease have high degree of toxicity. Thus, the aim of this study was to evaluate the effect of B. pilosa against T. gondii, by analyzing a total extract of this plant in parallel with a fraction obtained by precipitation in acetone. Also, it was assessed if the acetonic fraction could present lectinic activity, followed by its identification by mass spectrometry. It was observed with the experimental models designed that both total extract and acetonic fraction of B. pilosa were able to control T. gondii infection by in vitro and in vivo experiments, in addition to their low toxicity to host cells. Both total extract and acetonic fraction of this plant display capacity to impair replication of T. gondii tachyzoites. Interesting, the B. pilosa acetonic fraction treatment for 10 days after infection decreases significantly the number of T. gondii brain cyst in comparison with controls. The protein isolated from B. pilosa acetonic fraction was characterized as a novel lectin identified as maturase K. Taken together, these findings open new perspectives to treat patients infected by T. gondii. Future studies will be necessary to investigate the precise mechanism underlying the control of T. gondii infection to impair the replication of this parasite in the host cells after treatment with B. pilosa maturase K

Strategies used by Primary Care Nurses in the prevention of cervical cancer: integrative review

Objective: to identify, in the scientific literature, the strategies used by primary care nurses to prevent cervical cancer. Methods: An integrative review of the EMBASE, LILACS (BVS), SCOPUS, and Web of Science databases between March and April 2023. After the search, the selected articles were read in full, and the data were extracted for analysis. The final sample consisted of 5 studies. Results: the main strategies used were educational interventions, with a predominance of lectures and subsequent invitations or collection of material to carry out the preventive exam. Conversation circles (culture circles), telephone guidance, and the provision of a self-completed clinical form during the nursing consultation were also used. Conclusions: the studies made it possible to identify positive strategies for preventing cervical cancer, which could direct nurses to increasingly invest in methodologies aimed at increasing women’s autonomy. men

Um resgate histórico do LABIOMEC na história do CEFD 50 anos: parte 1

The purpose of this work is to report a little of the history and evolution that the Laboratory of Biomechanics (LABIOMEC) experienced over the 50 years of the Center for Physical Education and Sports (CEFD) of the Federal University of Santa Maria. For that purpose, searches were carried out on physical documents and digital files from 1970 to the present day in relation to LABIOMEC in articles, works in events, graduation monographs, specialization, dissertations and theses. The article was subdivided in two parts and in this first one, the historical recovery of the initial years is carried out. The importance of this report is to allow part of the history of LABIOMEC and the people who carried out their activities in the laboratory as teachers or students to be permanently registered.El propósito de este trabajo es informar un poco de la historia y evolución que experimentó el Laboratorio de Biomecánica (LABIOMEC) durante los 50 años del Centro de Educación Física y Deportes (CEFD) de la Universidad Federal de Santa María. Para ello, se realizaron búsquedas en documentos físicos y archivos digitales desde 1970 hasta la actualidad en relación con LABIOMEC en artículos, trabajos en eventos, monografías de graduación, especialización, disertaciones y tesis. El artículo fue subdividido en dos partes y em esta primera se realiza la recuperación histórica de los años iniciales y la evolución técnico-científica que se produjo. La importancia de esto informe es para permitir que parte de la historia de LABIOMEC y las personas que llevaron a cabo sus actividades en el laboratorio como maestros o estudiantes se registren permanentemente.Este trabalho tem como objetivo relatar um pouco da história e da evolução do Laboratório de Biomecânica (LABIOMEC) ao longo dos 50 anos do Centro de Educação Física e Desportos (CEFD) da Universidade Federal da Santa Maria. Para tanto foram realizadas buscas em documentos físicos e arquivos digitais no período de 1970 até os dias atuais em relação ao LABIOMEC em artigos, trabalhos em eventos, monografias de graduação, especialização, dissertações e teses. O artigo foi subdividido em duas partes e nesta primeira é realizado o resgate histórico dos anos iniciais e a evolução técnico-científica ocorrida. A importância desse relato é permitir que parte da história do LABIOMEC e das pessoas que realizaram suas atividades no laboratório como docentes ou discentes fique registrada de forma permanente