Metabolic reprogramming and lipid droplets are involved in Zika virus replication in neural cells.

Zika virus (ZIKV) infection is a global public health concern linked to adult neurological disorders and congenital diseases in newborns. Host lipid metabolism, including lipid droplet (LD) biogenesis, has been associated with viral replication and pathogenesis of different viruses. However, the mechanisms of LD formation and their roles in ZIKV infection in neural cells are still unclear. Here, we demonstrate that ZIKV regulates the expression of pathways associated with lipid metabolism, including the upregulation and activation of lipogenesis-associated transcription factors and decreased expression of lipolysis-associated proteins, leading to significant LD accumulation in human neuroblastoma SH-SY5Y cells and in neural stem cells (NSCs). Pharmacological inhibition of DGAT-1 decreased LD accumulation and ZIKV replication in vitro in human cells and in an in vivo mouse model of infection. In accordance with the role of LDs in the regulation of inflammation and innate immunity, we show that blocking LD formation has major roles in inflammatory cytokine production in the brain. Moreover, we observed that inhibition of DGAT-1 inhibited the weight loss and mortality induced by ZIKV infection in vivo. Our results reveal that LD biogenesis triggered by ZIKV infection is a crucial step for ZIKV replication and pathogenesis in neural cells. Therefore, targeting lipid metabolism and LD biogenesis may represent potential strategies for anti-ZIKV treatment development.

Dengue induces iNOS expression and nitric oxide synthesis in platelets through IL-1R.

Introduction: Dengue is an arthropod-born disease caused by dengue virus (DENV), that may manifest as a mild illness or severe form, characterized by hemorrhagic fever and shock. Nitric oxide (NO) is a vasodilator signaling molecule and an inhibitor of platelet aggregation known to be increased in platelets from dengue patients. However, the mechanisms underlying NO synthesis by platelets during dengue are not yet elucidated. IL-1ß is a pro-inflammatory cytokine able to induce iNOS expression in leukocytes and present in dengue patients at high levels. Nevertheless, the role of IL-1ß in platelet activation, especially regarding iNOS expression, are not clear. Methods: We prospectively followed a cohort of 28 dengue-infected patients to study NO synthesis in platelets and its relationship with disease outcomes. We used in vitro infection and stimulation models to gain insights on the mechanisms. Results and Discussion: We confirmed that platelets from dengue patients express iNOS and produce higher levels of NO during the acute phase compared to healthy volunteers, returning to normal levels after recovery. Platelet NO production during acute dengue infection was associated with the presence of warning signs, hypoalbuminemia and hemorrhagic manifestations, suggesting a role in dengue pathophysiology. By investigating the mechanisms, we evidenced increased iNOS expression in platelets stimulated with dengue patients´ plasma, indicating induction by circulating inflammatory mediators. We then investigated possible factors able to induce platelet iNOS expression and observed higher levels of IL-1ß in plasma from patients with dengue, which were correlated with NO production by platelets. Since platelets can synthesize and respond to IL-1ß, we investigated whether IL-1ß induces iNOS expression and NO synthesis in platelets. We observed that recombinant human IL-1ß enhanced iNOS expression and dose-dependently increased NO synthesis by platelets. Finally, platelet infection with DENV in vitro induced iNOS expression and NO production, besides the secretion of both IL-1α and IL-1ß. Importantly, treatment with IL-1 receptor antagonist or a combination of anti-IL-1α and anti-IL-1ß antibodies prevented DENV-induced iNOS expression and NO synthesis. Our data show that DENV induces iNOS expression and NO production in platelets through mechanisms depending on IL-1 receptor signaling.

Jóvenes universitarios de género masculino y el uso del preservativo / Jovens universitários do gênero masculino e a utilização do preservativo / Young male university students and condom use

Objetivo: Analizar el uso sistemático del condón por estudiantes universitarios varones. Método: Estudio transversal realizado en dos instituciones de educación superior de Río de Janeiro, con 661 estudiantes sexualmente activos de 18 a 29 años que respondieron un cuestionario. Los datos fueran analizados con el software Statistical Package for Social Sciences mediante estadística descriptiva e inferencial, con la aplicación de la prueba chi-cuadrado de Pearson. Resultados: Los datos muestran que los jóvenes son solteros (61,98%); heterosexuales (87,37%); no usan condones con parejas fijas (45,40%) o ocasionales (23,55%). El estado civil, la orientación sexual, las creencias religiosas, el consumo de alcohol y otras drogas antes de la última relación sexual, la percepción de riesgo y antecedentes de infecciones de transmisión sexual son un factor determinante para el uso del condón (valor de p <0,05). Conclusión: Considerando las características de la juventud y los factores socioculturales involucrados, se observa que los estudiantes asumen conductas de riesgo ante las infecciones de transmisión sexual con el uso no sistemático del condón. (AU)

Objective: to analyze systematic condom use by male university students. Method: A cross-sectional study carried out in two higher education institutions from Rio de Janeiro, with 661 sexually active students aged 18-29, who answered a questionnaire. The data were analyzed using the Statistical Package for Social Sciences software through descriptive and inferential statistics, with application of Pearson's chi-square test. Results: The data show that young people are single (61.98%), heterosexuals (87.37%) and that they do not use condoms with steady (45.40%) or occasional (23.55%) partners. Marital status, sexual orientation, religious belief, use of alcohol and other drugs before the last sexual relationship, risk perception and past history of sexually transmitted infections are determining factors for condom use (p-value<0.05). Conclusion: Considering the characteristics inherent to youth and the sociocultural factors involved, it is noted that the students assume risk behaviors in the face of sexually transmitted infections with non-systematic condom use. (AU)

Objetivo: Analisar o uso sistemático do preservativo por universitários do gênero masculino. Método: Estudo transversal realizado em duas instituições de ensino superior no Rio de Janeiro, com 661 estudantes de 18-29 anos, sexualmente ativos, que responderam um questionário. Os dados foram analisados com emprego do softwareStatistical Package for the Social Sciences através da estatística descritiva e inferencial, com a aplicação do teste qui-quadrado de Pearson. Resultados: Dados demonstram que os jovens são solteiros (61,98%); heterossexuais (87,37%); não usam preservativo com parcerias fixas (45,40%) ou eventuais (23,55%). A situação conjugal, orientação sexual, crença religiosa, uso de álcool e outras drogas antes da última relação sexual, percepção de risco e história pregressa de infecção sexualmente transmissível são fatores determinante para uso do preservativo (p-valor<0,05). Conclusão: Considerando as características próprias da juventude e fatores socioculturais envolvidos, nota-se que os estudantes assumem comportamentos de risco frente às infecções sexualmente transmissíveis com a utilização não sistemática do preservativo. (AU)

Ultrasensitive molecular tests for Plasmodium detection: applicability in control and elimination programs and reference laboratories.

Objective: To evaluate molecular tools to detect low-level parasitemia and the five species of Plasmodium that infect humans for use in control and elimination programs, and in reference laboratories. Methods: We evaluated 145 blood samples from patients who tested positive by nested polymerase chain reaction (nPCR), from asymptomatic individuals and from the WHO Global Malaria Programme/United Kingdom National External Quality Assessment Service. Samples were assayed using the genus-specific RealStar® Malaria PCR Kit 1.0 (alt-Gen; altona Diagnostics) and the RealStar® Malaria Screen & Type PCR Kit (alt-S&T; altona Diagnostics). The results from the molecular tests were compared with those from quantitative PCR (qPCR), nPCR and thick blood smear. Results: The levels of parasitemia ranged from 1 to 518 000 parasites/µL, depending on the species. Compared with nPCR, alt-S&T had a sensitivity of 100%, except for identifying P. falciparum, for which the sensitivity was 93.94%. All samples positive by alt-Gen were also positive by nPCR. When comparing alt-Gen to qPCR, the sensitivity was 100% for P. vivax, P. malariae and P. falciparum. For all Plasmodium species, the correlation between cycle threshold values of alt-S&T and alt-Gen compared with qPCR was significant (P < 0.0001, Spearman's test), with r = 0.8621 for alt-S&T and r = 0.9371 for alt-Gen. When all Plasmodium species were considered, there was a negative correlation between the level of parasitemia and real-time PCR cycle threshold values (P < 0.0001). In this study, only 2 of 28 samples from asymptomatic individuals were positive by thick blood smear; however, all 28 of these samples were positive by alt-S&T. Conclusions: The alt-Gen and alt-S&T assays are suitable for detecting submicroscopic infections for distinct epidemiological purposes, such as for use in surveys and reference laboratories, and screening in blood banks, which will contribute to global efforts to eliminate malaria.

Ultrasensitive molecular tests for Plasmodium detection: applicability in control and elimination programs and reference laboratories

[ABSTRACT]. Objective. To evaluate molecular tools to detect low-level parasitemia and the five species of Plasmodium that infect humans for use in control and elimination programs, and in reference laboratories. Methods. We evaluated 145 blood samples from patients who tested positive by nested polymerase chain reaction (nPCR), from asymptomatic individuals and from the WHO Global Malaria Programme/United Kingdom National External Quality Assessment Service. Samples were assayed using the genus-specific RealStar® Malaria PCR Kit 1.0 (alt-Gen; altona Diagnostics) and the RealStar® Malaria Screen & Type PCR Kit (alt-S&T; altona Diagnostics). The results from the molecular tests were compared with those from quantitative PCR (qPCR), nPCR and thick blood smear. Results. The levels of parasitemia ranged from 1 to 518 000 parasites/μL, depending on the species. Compared with nPCR, alt-S&T had a sensitivity of 100%, except for identifying P. falciparum, for which the sensitivity was 93.94%. All samples positive by alt-Gen were also positive by nPCR. When comparing alt-Gen to qPCR, the sensitivity was 100% for P. vivax, P. malariae and P. falciparum. For all Plasmodium species, the correlation between cycle threshold values of alt-S&T and alt-Gen compared with qPCR was significant (P < 0.0001, Spearman’s test), with r = 0.8621 for alt-S&T and r = 0.9371 for alt-Gen. When all Plasmodium species were considered, there was a negative correlation between the level of parasitemia and real-time PCR cycle threshold values (P < 0.0001). In this study, only 2 of 28 samples from asymptomatic individuals were positive by thick blood smear; however, all 28 of these samples were positive by alt-S&T. Conclusions. The alt-Gen and alt-S&T assays are suitable for detecting submicroscopic infections for distinct epidemiological purposes, such as for use in surveys and reference laboratories, and screening in blood banks, which will contribute to global efforts to eliminate malaria.

[RESUMEN]. Objetivo. Evaluar herramientas moleculares para detectar bajos niveles de parasitemia y las cinco especies de Plasmodium que infectan a los seres humanos, a fin de emplearlas en los programas de control y eliminación y en los laboratorios de referencia. Métodos. Se evaluaron 145 muestras de sangre de pacientes positivos por reacción en cadena de la polimerasa anidada (nPCR), de individuos asintomáticos y de muestras del Programa Mundial de Malaria de la Organización Mundial de la Salud/Servicio Nacional de Evaluación Externa de Calidad del Reino Unido. Las muestras se analizaron con el kit de PCR RealStar® Malaria 1.0 (alt-Gen; altona Diagnostics), específico para cada género, y con el kit de PCR RealStar® Malaria Screen & Type (alt-S&T; altona Diagnostics). Se compararon los resultados de las pruebas moleculares con los de la PCR cuantitativa (qPCR), la nPCR y el frotis de gota gruesa. Resultados. Los niveles de parasitemia oscilaron entre 1 y 518 000 parásitos/μl, según la especie. En comparación con la nPCR, la prueba alt-S&T tuvo una sensibilidad del 100%, excepto para la identificación de P. falciparum, para el cual la sensibilidad fue del 93,94%. Todas las muestras positivas por alt-Gen lo fueron también por nPCR. Al comparar alt-Gen con la qPCR, la sensibilidad fue del 100% para P. vivax, P. malariae y P. falciparum. Para todas las especies de Plasmodium, la correlación entre los valores del umbral de ciclo de alt-S&T y alt-Gen en comparación con la qPCR fue significativa (P < 0,0001, prueba de Spearman), con r = 0,8621 para alt-S&T y r = 0,9371 para alt-Gen. Cuando se consideraron todas las especies de Plasmodium hubo una correlación negativa entre el nivel de parasitemia y los valores de umbral de ciclo de PCR en tiempo real (P < 0,0001). En este estudio, solo 2 de las 28 muestras de individuos asintomáticos fueron positivas por frotis de gota gruesa; sin embargo, las 28 muestras fueron positivas por alt-S&T. Conclusiones. Los ensayos alt-Gen y alt-S&T son adecuados para detectar infecciones submicroscópicas con distintos fines epidemiológicos, como su uso en investigaciones y laboratorios de referencia y el cribado en bancos de sangre, lo que contribuirá a los esfuerzos mundiales para eliminar la malaria.

[RESUMO]. Objectivo. Avaliar ferramentas moleculares para detectar parasitemia de baixo nível e as cinco espécies de Plasmodium que infectam humanos, para utilização em programas de controlo e eliminação e em laboratórios de referência. Métodos. Avaliámos 145 amostras de sangue de doentes que testaram positivo por reacção em cadeia da polimerase aninhada (nPCR), de indivíduos assintomáticos, e do Programa Global de Paludismo da Organização Mundial de Saúde/Serviço Nacional de Avaliação da Qualidade Externa do Reino Unido. As amostras foram ensaiadas utilizando o RealStar® Malaria PCR Kit 1.0 (alt-Gen; altona Diagnostics) e o RealStar® Malaria Screen & Type PCR Kit (alt-S&T; altona Diagnostics). Os resultados dos testes moleculares foram comparados com os resultados da PCR quantitativa (qPCR), nPCR e exame da gota espessa. Resultados. Os níveis de parasitemia variaram de 1 a 518 000 parasitas/μL, dependendo da espécie. Em comparação com a nPCR, alt-S&T tinha uma sensibilidade de 100%, excepto na identificação de P. falciparum, para a qual a sensibilidade era de 93,94%. Todas as amostras positivas por alt-Gen foram também positivas por nPCR. Ao comparar alt-Gen com qPCR, a sensibilidade foi de 100% para P. vivax, P. malariae e P. falciparum. Para todas as espécies Plasmodium, a correlação entre os valores limiares de ciclo de alt-S&T e alt-Gen comparados com qPCR foi significativa (P < 0,0001, teste de Spearman), com r = 0,8621 para alt- S&T e r = 0,9371 para alt-Gen. Quando todas as espécies de Plasmodium foram consideradas, houve uma correlação negativa entre o nível de parasitemia e os valores limiares do ciclo de PCR em tempo real (P < 0,0001). Neste estudo, apenas 2 de 28 amostras de indivíduos assintomáticos foram positivas por exame da gota espessa; no entanto, todas estas 28 amostras foram positivas por alt-S&T. Conclusões. Os ensaios alt-Gen e alt-S&T são adequados para a detecção de infecções submicroscópicas para fins epidemiológicos distintos, tais como para utilização em inquéritos e laboratórios de referência e o rastreio em bancos de sangue, o que contribuirá para os esforços globais de eliminação da malária.

Ultrasensitive molecular tests for Plasmodium detection: applicability in control and elimination programs and reference laboratories / Pruebas moleculares ultrasensibles para la detección de Plasmodium: aplicabilidad en los programas de control y eliminación y en los laboratorios de referencia / Testes moleculares ultra-sensíveis para detecção de Plasmodium: aplicabilidade em programas de controlo e eliminação e laboratórios de referencia

ABSTRACT Objective. To evaluate molecular tools to detect low-level parasitemia and the five species of Plasmodium that infect humans for use in control and elimination programs, and in reference laboratories. Methods. We evaluated 145 blood samples from patients who tested positive by nested polymerase chain reaction (nPCR), from asymptomatic individuals and from the WHO Global Malaria Programme/United Kingdom National External Quality Assessment Service. Samples were assayed using the genus-specific RealStar® Malaria PCR Kit 1.0 (alt-Gen; altona Diagnostics) and the RealStar® Malaria Screen & Type PCR Kit (alt-S&T; altona Diagnostics). The results from the molecular tests were compared with those from quantitative PCR (qPCR), nPCR and thick blood smear. Results. The levels of parasitemia ranged from 1 to 518 000 parasites/µL, depending on the species. Compared with nPCR, alt-S&T had a sensitivity of 100%, except for identifying P. falciparum, for which the sensitivity was 93.94%. All samples positive by alt-Gen were also positive by nPCR. When comparing alt-Gen to qPCR, the sensitivity was 100% for P. vivax, P. malariae and P. falciparum. For all Plasmodium species, the correlation between cycle threshold values of alt-S&T and alt-Gen compared with qPCR was significant (P < 0.0001, Spearman's test), with r = 0.8621 for alt-S&T and r = 0.9371 for alt-Gen. When all Plasmodium species were considered, there was a negative correlation between the level of parasitemia and real-time PCR cycle threshold values (P < 0.0001). In this study, only 2 of 28 samples from asymptomatic individuals were positive by thick blood smear; however, all 28 of these samples were positive by alt-S&T. Conclusions. The alt-Gen and alt-S&T assays are suitable for detecting submicroscopic infections for distinct epidemiological purposes, such as for use in surveys and reference laboratories, and screening in blood banks, which will contribute to global efforts to eliminate malaria.

RESUMEN Objetivo. Evaluar herramientas moleculares para detectar bajos niveles de parasitemia y las cinco especies de Plasmodium que infectan a los seres humanos, a fin de emplearlas en los programas de control y eliminación y en los laboratorios de referencia. Métodos. Se evaluaron 145 muestras de sangre de pacientes positivos por reacción en cadena de la polimerasa anidada (nPCR), de individuos asintomáticos y de muestras del Programa Mundial de Malaria de la Organización Mundial de la Salud/Servicio Nacional de Evaluación Externa de Calidad del Reino Unido. Las muestras se analizaron con el kit de PCR RealStar® Malaria 1.0 (alt-Gen; altona Diagnostics), específico para cada género, y con el kit de PCR RealStar® Malaria Screen & Type (alt-S&T; altona Diagnostics). Se compararon los resultados de las pruebas moleculares con los de la PCR cuantitativa (qPCR), la nPCR y el frotis de gota gruesa. Resultados. Los niveles de parasitemia oscilaron entre 1 y 518 000 parásitos/µl, según la especie. En comparación con la nPCR, la prueba alt-S&T tuvo una sensibilidad del 100%, excepto para la identificación de P. falciparum, para el cual la sensibilidad fue del 93,94%. Todas las muestras positivas por alt-Gen lo fueron también por nPCR. Al comparar alt-Gen con la qPCR, la sensibilidad fue del 100% para P. vivax, P. malariae y P. falciparum. Para todas las especies de Plasmodium, la correlación entre los valores del umbral de ciclo de alt-S&T y alt-Gen en comparación con la qPCR fue significativa (P < 0,0001, prueba de Spearman), con r = 0,8621 para alt-S&T y r = 0,9371 para alt-Gen. Cuando se consideraron todas las especies de Plasmodium hubo una correlación negativa entre el nivel de parasitemia y los valores de umbral de ciclo de PCR en tiempo real (P < 0,0001). En este estudio, solo 2 de las 28 muestras de individuos asintomáticos fueron positivas por frotis de gota gruesa; sin embargo, las 28 muestras fueron positivas por alt-S&T. Conclusiones. Los ensayos alt-Gen y alt-S&T son adecuados para detectar infecciones submicroscópicas con distintos fines epidemiológicos, como su uso en investigaciones y laboratorios de referencia y el cribado en bancos de sangre, lo que contribuirá a los esfuerzos mundiales para eliminar la malaria.

RESUMO Objectivo. Avaliar ferramentas moleculares para detectar parasitemia de baixo nível e as cinco espécies de Plasmodium que infectam humanos, para utilização em programas de controlo e eliminação e em laboratórios de referência. Métodos. Avaliámos 145 amostras de sangue de doentes que testaram positivo por reacção em cadeia da polimerase aninhada (nPCR), de indivíduos assintomáticos, e do Programa Global de Paludismo da Organização Mundial de Saúde/Serviço Nacional de Avaliação da Qualidade Externa do Reino Unido. As amostras foram ensaiadas utilizando o RealStar® Malaria PCR Kit 1.0 (alt-Gen; altona Diagnostics) e o RealStar® Malaria Screen & Type PCR Kit (alt-S&T; altona Diagnostics). Os resultados dos testes moleculares foram comparados com os resultados da PCR quantitativa (qPCR), nPCR e exame da gota espessa. Resultados. Os níveis de parasitemia variaram de 1 a 518 000 parasitas/µL, dependendo da espécie. Em comparação com a nPCR, alt-S&T tinha uma sensibilidade de 100%, excepto na identificação de P. falciparum, para a qual a sensibilidade era de 93,94%. Todas as amostras positivas por alt-Gen foram também positivas por nPCR. Ao comparar alt-Gen com qPCR, a sensibilidade foi de 100% para P. vivax, P. malariae e P. falciparum. Para todas as espécies Plasmodium, a correlação entre os valores limiares de ciclo de alt-S&T e alt-Gen comparados com qPCR foi significativa (P < 0,0001, teste de Spearman), com r = 0,8621 para alt-S&T e r = 0,9371 para alt-Gen. Quando todas as espécies de Plasmodium foram consideradas, houve uma correlação negativa entre o nível de parasitemia e os valores limiares do ciclo de PCR em tempo real (P < 0,0001). Neste estudo, apenas 2 de 28 amostras de indivíduos assintomáticos foram positivas por exame da gota espessa; no entanto, todas estas 28 amostras foram positivas por alt-S&T. Conclusões. Os ensaios alt-Gen e alt-S&T são adequados para a detecção de infecções submicroscópicas para fins epidemiológicos distintos, tais como para utilização em inquéritos e laboratórios de referência e o rastreio em bancos de sangue, o que contribuirá para os esforços globais de eliminação da malária.

Practices for the prevention of sexually transmitted infections among university students / Prácticas para la prevención de infecciones de transmisión sexual entre estudiantes universitarios / Práticas de prevenção de infecções sexualmente transmissíveis entre estudantes universitários

ABSTRACT Objective: to analyze the sexual practices adopted by university students for the prevention of Sexually Transmitted Infections Method: a quantitative, descriptive and cross-sectional study, conducted in 2016, at a private university in the municipality of Rio de Janeiro, Brazil. A convenience sample was selected, stratified by gender, of 768 students who answered a self-administered questionnaire, structured with variables of sociodemographic characterization and related to knowledge about sexually transmitted infections, sexual practices, prevention practices and care with sexual health. In the analysis, descriptive statistics, chi-square tests and analysis of variance were used, with a significance level of 5%. Results: most of the university students ‒ 654 (85.16%) ‒ had an active sex life and 480 (62.54%) did not use condoms in all their sexual encounters. Among the participants, 509 (84.83%) said they had sexual intercourse with a steady partner, of which 224 (44.01%) used a condom. In the investigated group, 313 (47.86%) had relationships with casual partners, with 199 (63.58%) reporting having used a condom. Among the participants, 174 (26.61%) had their sexual practices classified as adequate/satisfactory. Conclusion: the findings show that the university students investigated present a risk behavior for Sexually Transmitted Infections due to inadequate/unsatisfactory sexual practices. Health education actions should consider cultural and individual aspects of the group in order to encourage reflection on practices for the prevention of sexually transmitted diseases.

RESUMEN Objetivo: analizar las prácticas sexuales adoptadas por estudiantes universitarios para prevenir Infecciones de Transmisión Sexual. Método: estudio cuantitativo, descriptivo y transversal, realizado en el año 2016 en una universidad privada del municipio de Río de Janeiro, Brasil. Ajustada por conveniencia y estratificada por sexo, la muestra estuvo compuesta por 768 estudiantes que respondieron un cuestionario autoaplicado, estructurado con variables de caracterización sociodemográfica y relacionadas con el conocimiento sobre infecciones de transmisión sexual, prácticas sexuales, prácticas de prevención y cuidados con la salud sexual. En el análisis se empleó la estadística descriptiva, pruebas de Chi-cuadrado y análisis de variancia, con un nivel de significancia del 5%. Resultados: la mayoría de los universitarios ‒ 654 (85,16%) ‒ tenía una vida sexual activa y 480 (62,54%) no usaban preservativo en todos sus encuentros sexuales. Entre los participantes, 509 (84,83%) afirmaron tener relaciones sexuales con una pareja fija y, de ellos, 224 (44,01%) utilizaban preservativo. En el grupo investigado, 313 (47,86%) tuvieron relaciones con parejas casuales, y 199 (63,58%) declararon haber usado preservativo. Entre los participantes, las prácticas sexuales de 174 (26,61%) de ellos se clasificaron como adecuadas/satisfactorias. Conclusión: las conclusiones del estudio evidencian que los universitarios investigados presentan un comportamiento de riesgo para contraer Infecciones de Transmisión Sexual como resultado de prácticas sexuales inadecuadas/insatisfactorias. Las medidas de educación en salud deben considerar aspectos culturales e individuales del grupo para favorecer la reflexión sobre las prácticas de prevención de enfermedades de transmisión sexual.

RESUMO Objetivo: analisar as práticas sexuais adotadas por estudantes universitários para prevenção de Infecções Sexualmente Transmissíveis Método: estudo quantitativo, descritivo e transversal, realizado em 2016, em uma universidade privada no município do Rio de Janeiro, Brasil. Selecionou-se amostra por conveniência, estratificada por sexo, de 768 estudantes que responderam a um questionário autoaplicado, estruturado com variáveis de caracterização sociodemográfica e relacionadas ao conhecimento sobre infecções sexualmente transmissíveis, práticas sexuais, práticas de prevenção e cuidados com a saúde sexual. Na análise, empregou-se a estatística descritiva, testes de quiquadrado e análise de variância, com nível de significância de 5%. Resultados: a maioria dos universitários ‒ 654(85,16%) ‒ tinha vida sexual ativa e 480(62,54%) não fazia uso do preservativo em todos os intercursos sexuais. Entre os participantes, 509(84,83%) afirmaram ter relação sexual com parceiro fixo, dos quais 224(44,01%) utilizaram o preservativo. No grupo investigado, 313(47,86%) tiveram relações com parceiros casuais, sendo que 199 (63,58%) informaram ter usado o preservativo. Entre os participantes, 174(26,61%) tiveram suas práticas sexuais classificadas como adequadas/satisfatórias. Conclusão: os achados evidenciam que os universitários investigados apresentam um comportamento de risco para Infecções Sexualmente Transmissíveis decorrente de práticas sexuais inadequadas/insatisfatórias. Ações de educação em saúde devem considerar aspectos culturais e individuais do grupo para favorecer a reflexão sobre as práticas de prevenção de doenças transmitidas pelo sexo.

Alterações posturais: a caracterização de discentes de Fisioterapia em uma instituição do ensino superior em Fortaleza/CE / Postural changes: the characterization of Physiotherapy students in a higher education institution in Fortaleza/CE

Na postura padrão, a coluna apresenta curvaturas normais e os ossos dos membros inferiores ficam em alinhamento. Quando o componente estrutural é alterado, o corpo humano modifica-se para desempenhar tal situação da melhor forma possível, o que pode ocasionar em um desvio postural. O objetivo desta pesquisa é avaliar a postura e seus comprometimentos em discentes de um curso de Fisioterapia, traçando o perfil clínico socioeconômico, conhecendo os comprometimentos posturais dos participantes da pesquisa. Tratou-se de um estudo transversal, descritivo e quantitativo realizado entre os meses de agosto de 2015 a junho de 2016, no Centro Universitário Estácio do Ceará com 32 discentes do curso de Fisioterapia. Os dados foram coletados através de uma ficha de avaliação postural desenvolvida para o inquérito e analisada no programa Excel® 2013 e confrontados com a literatura vigente sobre o assunto. O gênero feminino foi predominante em 75% da amostra. Em relação ao estado civil dos participantes, 87,5% eram solteiros. A média de idade foi de 24,4 anos. Em relação ao Índice de Massa Corpórea, foi evidenciado que 28 participantes (87,5%) apresentaram um peso normal, de acordo com sua altura e peso. Para verificar a presença de escoliose, foi realizado o Teste de Adams. Evidenciou-se que 72% dos participantes apresentaram gibosidade, 12,50% apresentaram gibosidade lombar, 43,75% apresentaram gibosidade torácica e 15,63% apresentaram gibosidade toracolombar. A maioria dos estudantes apresentou escoliose. No entanto, estudos com uma amostra maior devem ser realizados, a fim de que resultados mais abrangentes possam ser obtidos. (AU)

In the standard posture, the spine has normal curvatures and the bones of the lower limbs are in alignment. When the structural component is changed, the human body changes to perform this situation in the best possible way, which can cause a postural deviation. The objective of this study was to evaluate the posture and its commitments in students of a Physical therapy course, tracing the socioeconomic clinical profile, knowing the postural impairments of the participants. This was a cross-sectional, descriptive and quantitative study carried out between August 2015 and June 2016, at the Estácio do Ceará University Center with 32 students. The data were collected through a postural assessment form developed for the survey and analyzed using the Excel® program and compared with the current literature. The female gender was predominant in 75% of the sample. Regarding the participants' marital status, 87.5% were single. The average age was 24.4 years. Regarding the Body Mass Index, it was evidenced that 28 participants (87.5%) had a normal weight, according to their height. To test for the presence of scoliosis, the Adams Test was performed. 72% of the participants presented gibosity, 12.50% lumbar gibosity, 43.75% thoracic gibosity and 15.63% thoraco-lumbar gibosity. Most of the students had scoliosis. However, studies with a larger sample should be carried out to obtain more accurate results. (AU)

Dengue virus-activated platelets modulate monocyte immunometabolic response through lipid droplet biogenesis and cytokine signaling.

Dengue is characterized as one of the most important arthropod-borne human viral diseases, representing a public health problem. Increased activation of immune cells is involved in the progression of infection to severe forms. Recently, our group demonstrated the contribution of platelet-monocyte interaction to inflammatory responses in dengue, adding to evolving evidence that platelets have inflammatory functions and can regulate different aspects of innate immune responses. Furthermore, stimuli-specific-activated platelets can promote phenotypic changes and metabolic reprogramming in monocytes. Thus, this study aimed to evaluate the roles of dengue virus (DENV)-activated platelets on immunometabolic reprogramming of monocytes in vitro, focusing on lipid droplet (LD) biogenesis. We demonstrated that platelets exposed to DENV in vitro form aggregates with monocytes and signal to LD formation and CXCL8/IL-8, IL-10, CCL2, and PGE2 secretion. Pharmacologic inhibition of LD biogenesis prevents PGE2 secretion, but not CXCL8/IL-8 release, by platelet-monocyte complexes. In exploring the mechanisms involved, we demonstrated that LD formation in monocytes exposed to DENV-activated platelets is partially dependent on platelet-produced MIF. Additionally, LD formation is higher in monocytes, which have platelets adhered on their surface, suggesting that beyond paracrine signaling, platelet adhesion is an important event in platelet-mediated modulation of lipid metabolism in monocytes. Together, our results demonstrate that activated platelets aggregate with monocytes during DENV infection and signal to LD biogenesis and the secretion of inflammatory mediators, which may contribute to dengue immunopathogenesis.

Identification of plasmid IncQ1 and NTEKPC-IId harboring bla KPC-2 in isolates from Klebsiella pneumoniae infections in patients from Recife-PE, Brazil.

INTRODUCTION: This study investigated the genetic environment of bla KPC-2 in Klebsiella pnemoniae multi-drug resistant clinical isolates. METHODS: Four carbapenemase gene isolates resistant to carbapenems, collected from infected patients from two hospitals in Brazil, were investigated using polymerase chain reaction and plasmid DNA sequencing. RESULTS: The bla KPC-2 gene was located between ISKpn6 and a resolvase tnpR in the non-Tn4401 element (NTEKPC-IId). It was detected on a plasmid belonging to the IncQ1 group. CONCLUSIONS: To our knowledge, this is the first report of the presence of the bla KPC-2 gene in the NTEKPC-IId element carried by plasmid IncQ1 from infections in Brazil.

Inflammatory signaling in dengue-infected platelets requires translation and secretion of nonstructural protein 1.

Emerging evidence identifies major contributions of platelets to inflammatory amplification in dengue, but the mechanisms of infection-driven platelet activation are not completely understood. Dengue virus nonstructural protein-1 (DENV NS1) is a viral protein secreted by infected cells with recognized roles in dengue pathogenesis, but it remains unknown whether NS1 contributes to the inflammatory phenotype of infected platelets. This study shows that recombinant DENV NS1 activated platelets toward an inflammatory phenotype that partially reproduced DENV infection. NS1 stimulation induced translocation of α-granules and release of stored factors, but not of newly synthesized interleukin-1ß (IL-1ß). Even though both NS1 and DENV were able to induce pro-IL-1ß synthesis, only DENV infection triggered caspase-1 activation and IL-1ß release by platelets. A more complete thromboinflammatory phenotype was achieved by synergistic activation of NS1 with classic platelet agonists, enhancing α-granule translocation and inducing thromboxane A2 synthesis (thrombin and platelet-activating factor), or activating caspase-1 for IL-1ß processing and secretion (adenosine triphosphate). Also, platelet activation by NS1 partially depended on toll-like receptor-4 (TLR-4), but not TLR-2/6. Finally, the platelets sustained viral genome translation and replication, but did not support the release of viral progeny to the extracellular milieu, characterizing an abortive viral infection. Although DENV infection was not productive, translation of the DENV genome led to NS1 expression and release by platelets, contributing to the activation of infected platelets through an autocrine loop. These data reveal distinct, new mechanisms for platelet activation in dengue, involving DENV genome translation and NS1-induced platelet activation via platelet TLR4.

Is it possible to substitute the monofilament test for the Ipswich Touch Test in screening for peripheral diabetic neuropathy?

BACKGROUND: This study aimed to assess the agreement and efficacy of the Ipswich Touch Test compared to the monofilament test in individuals with type 2 diabetes. MATERIALS AND METHODS: A cross-sectional and analytical study was conducted. The inclusion criteria were patients with type II diabetes (n = 250) who did not present ulcers or amputation in either foot. The exclusion criteria were as follows: patients who presented sequelae of cerebrovascular disease or other neurological pathologies, as well as diagnoses of malignancy, alcohol abuse, liver cirrhosis, hepatitis B, AIDS, hypothyroidism, chronic kidney disease or lupus erythaematosus, as these clinical conditions could influence or bias the results (Won and Park in Endocrinol Metab 31:230-238, 2016). Sensitivity, specificity, predictive values, likelihood ratios, and Kappa index were calculated. Other factors assessed were glycated haemoglobin and body mass index. RESULTS: Most of the participants were female (71.2%), and glycated haemoglobin (HbA1c) was greater than 7% in 54.4% of the patients. The mean age was 59.43 years, and the mean time since diagnosis was 12.38 years. The Kappa index was 0.819 (p < 0.001), and the Ipswich Touch Test had a sensitivity of 83.33%, a specificity of 97.66%, a positive predictive value of 85.71%, a negative predictive value of 97.21%, a positive likelihood ratio of 30.19%, and a negative likelihood ratio of 0.17%. The level of significance was 5% in this study. CONCLUSION: The Ipswich Touch Test resented good agreement and efficacy compared to the gold standard-the 10 g monofilament test.

Identification of plasmid IncQ1 and NTEKPC-IId harboring bla KPC-2 in isolates from Klebsiella pneumoniae infections in patients from Recife-PE, Brazil

Abstract INTRODUCTION: This study investigated the genetic environment of bla KPC-2 in Klebsiella pnemoniae multi-drug resistant clinical isolates. METHODS: Four carbapenemase gene isolates resistant to carbapenems, collected from infected patients from two hospitals in Brazil, were investigated using polymerase chain reaction and plasmid DNA sequencing. RESULTS: The bla KPC-2 gene was located between ISKpn6 and a resolvase tnpR in the non-Tn4401 element (NTEKPC-IId). It was detected on a plasmid belonging to the IncQ1 group. CONCLUSIONS To our knowledge, this is the first report of the presence of the bla KPC-2 gene in the NTEKPC-IId element carried by plasmid IncQ1 from infections in Brazil.

Emergence of the East-Central-South-African genotype of Chikungunya virus in Brazil and the city of Rio de Janeiro may have occurred years before surveillance detection.

Brazil, which is hyperendemic for dengue virus (DENV), has had recent Zika (ZIKV) and (CHIKV) Chikungunya virus outbreaks. Since March 2016, CHIKV is the arbovirus infection most frequently diagnosed in Rio de Janeiro. In the analysis of 1835 syndromic patients, screened by real time RT-PCR, 56.4% of the cases were attributed to CHIKV, 29.6% to ZIKV, and 14.1% to DENV-4. Sequence analyses of CHIKV from sixteen samples revealed that the East-Central-South-African (ECSA) genotype of CHIKV has been circulating in Brazil since 2013 [95% bayesian credible interval (BCI): 03/2012-10/2013], almost a year before it was detected by arbovirus surveillance program. Brazilian cases are related to Central African Republic sequences from 1980's. To the best of our knowledge, given the available sequence published here and elsewhere, the ECSA genotype was likely introduced to Rio de Janeiro early on 2014 (02/2014; BCI: 07/2013-08/2014) through a single event, after primary circulation in the Bahia state at the Northestern Brazil in the previous year. The observation that the ECSA genotype of CHIKV was circulating undetected underscores the need for improvements in molecular methods for viral surveillance.

Capturing sequence diversity in metagenomes with comprehensive and scalable probe design.

Metagenomic sequencing has the potential to transform microbial detection and characterization, but new tools are needed to improve its sensitivity. Here we present CATCH, a computational method to enhance nucleic acid capture for enrichment of diverse microbial taxa. CATCH designs optimal probe sets, with a specified number of oligonucleotides, that achieve full coverage of, and scale well with, known sequence diversity. We focus on applying CATCH to capture viral genomes in complex metagenomic samples. We design, synthesize, and validate multiple probe sets, including one that targets the whole genomes of the 356 viral species known to infect humans. Capture with these probe sets enriches unique viral content on average 18-fold, allowing us to assemble genomes that could not be recovered without enrichment, and accurately preserves within-sample diversity. We also use these probe sets to recover genomes from the 2018 Lassa fever outbreak in Nigeria and to improve detection of uncharacterized viral infections in human and mosquito samples. The results demonstrate that CATCH enables more sensitive and cost-effective metagenomic sequencing.

Yellow fever virus is susceptible to sofosbuvir both in vitro and in vivo.

Yellow fever virus (YFV) is a member of the Flaviviridae family. In Brazil, yellow fever (YF) cases have increased dramatically in sylvatic areas neighboring urban zones in the last few years. Because of the high lethality rates associated with infection and absence of any antiviral treatments, it is essential to identify therapeutic options to respond to YFV outbreaks. Repurposing of clinically approved drugs represents the fastest alternative to discover antivirals for public health emergencies. Other Flaviviruses, such as Zika (ZIKV) and dengue (DENV) viruses, are susceptible to sofosbuvir, a clinically approved drug against hepatitis C virus (HCV). Our data showed that sofosbuvir docks onto YFV RNA polymerase using conserved amino acid residues for nucleotide binding. This drug inhibited the replication of both vaccine and wild-type strains of YFV on human hepatoma cells, with EC50 values around 5 µM. Sofosbuvir protected YFV-infected neonatal Swiss mice and adult type I interferon receptor knockout mice (A129-/-) from mortality and weight loss. Because of its safety profile in humans and significant antiviral effects in vitro and in mice, Sofosbuvir may represent a novel therapeutic option for the treatment of YF. Key-words: Yellow fever virus; Yellow fever, antiviral; sofosbuvir.

Beyond Members of the Flaviviridae Family, Sofosbuvir Also Inhibits Chikungunya Virus Replication.

Chikungunya virus (CHIKV) causes a febrile disease associated with chronic arthralgia, which may progress to neurological impairment. Chikungunya fever (CF) is an ongoing public health problem in tropical and subtropical regions of the world, where control of the CHIKV vector, Aedes mosquitos, has failed. As there is no vaccine or specific treatment for CHIKV, patients receive only palliative care to alleviate pain and arthralgia. Thus, drug repurposing is necessary to identify antivirals against CHIKV. CHIKV RNA polymerase is similar to the orthologue enzyme of other positive-sense RNA viruses, such as members of the Flaviviridae family. Among the Flaviviridae, not only is hepatitis C virus RNA polymerase susceptible to sofosbuvir, a clinically approved nucleotide analogue, but so is dengue, Zika, and yellow fever virus replication. Here, we found that sofosbuvir was three times more selective in inhibiting CHIKV production in human hepatoma cells than ribavirin, a pan-antiviral drug. Although CHIKV replication in human induced pluripotent stem cell-derived astrocytes was less susceptible to sofosbuvir than were hepatoma cells, sofosbuvir nevertheless impaired virus production and cell death in a multiplicity of infection-dependent manner. Sofosbuvir also exhibited antiviral activity in vivo by preventing CHIKV-induced paw edema in adult mice at a dose of 20 mg/kg of body weight/day and prevented mortality in a neonate mouse model at 40- and 80-mg/kg/day doses. Our data demonstrate that a prototypic alphavirus, CHIKV, is also susceptible to sofosbuvir. As sofosbuvir is a clinically approved drug, our findings could pave the way to it becoming a therapeutic option against CF.

Práticas de prevenção de infecções sexualmente transmissíveis de universitários do sexo masculino: estudo comparativo / Sexually Transmitted Infections prevention practices of male college students: a comparative study

O estudo teve como objetivo geral: analisar comparativamente as práticas de prevenção das infecções sexualmente transmissíveis (IST) de jovens universitários do sexo masculino de duas instituições. E como objetivos específicos: identificar as práticas sexuais dos jovens universitários frente às IST; caracterizar as práticas de prevenção de IST adotadas pelos universitários; comparar as práticas sexuais e de prevenção de IST adotadas por eles. Estudo quantitativo, descritivo, transversal, desenvolvido em duas instituições de ensino superior (IES1 e IES2), no Rio de Janeiro, com amostra do tipo intencional e estratificada, de 768 jovens universitários do sexo masculino de 18 a 29 anos. Para captação das informações foi utilizado um questionário estruturado com 60 questões. A análise dos dados empregou a estatística descritiva, em frequências absolutas, relativas e análise bivariada; para a análise inferencial foi utilizado o Software Statistical Package for the Social Sciences (SPSS) e aplicado o teste qui-quadrado de Pearson. Os resultados evidenciaram que os universitários das duas instituições apresentam práticas sexuais e comportamento de risco semelhante para aquisição de IST, havendo poucas diferenças. A maioria é sexualmente ativa, tendo tido o primeiro intercurso sexual e usado o preservativo nessa ocasião. Um quantitativo expressivo de participantes não utiliza preservativo sempre e não costuma negociar o seu uso. No tocante ao emprego do preservativo com parcerias fixas, os universitários da IES1 apresentaram resultados em quantitativo maior que os participantes da IES2. Em relação ao conhecimento sobre as IST, embora os jovens tenham informado que não conhecem o suficiente, reconhecem algum método para a prevenção das infecções. As parcerias sexuais dos participantes não costumam usar o preservativo feminino. O uso de álcool e/ou drogas foi informado pela maioria dos jovens, contudo o seu emprego não precedeu à última relação sexual. Nas duas instituições, grande parte não utiliza o sistema público de assistência à saúde, nunca realizou teste para detecção do vírus da imunodeficiência humana (HIV) e não são circuncidados. Conclui-se que os participantes do estudo assumem um comportamento de risco e ficam vulneráveis às IST e, nesse contexto são oportunas ações educativas que valorizem o autocuidado de jovens e orientações relacionadas à prevenção de agravos à saúde sexual. É preciso que os profissionais envolvidos no atendimento a população jovem sejam capacitados e, estimulem a reflexão sobre a importância dos cuidados com a saúde sexual, com destaque para o público masculino que não busca com frequência os serviços de saúde, adesão às práticas de prevenção de agravos e assunção de um comportamento de risco.

The general objective of the study was to analyze sexually transmitted infections (STI) of young male college students from two institutions. And as specific objectives: to identify the sexual practices of college students against STI; to characterize the practices of prevention of STI adopted by university students; to compare the sexual practices and prevention of STI adopted by them. A quantitative, descriptive, cross-sectional study developed in two higher education institutions (IES1 and IES2), in Rio de Janeiro, with a sample of the intentional and stratified type, of 768 university students, male, aged 18 to 29 years. A questionnaire structured with 60 questions was used to gather information. Data analysis employed descriptive statistics, in absolute, relative frequencies and bivariate analysis; for the inferential analysis we used the Statistical Package for the Social Sciences (SPSS) and applied the chi- square test of Pearson. The results showed that the students of the two institutions presented sexual practices and similar risk behavior for STI acquisition, with few differences. Most are sexually active, having had their first sexual intercourse and using condoms on this occasion. An expressive number of participants does not use condoms at all and does not usually negotiate their use. Regarding the use of the condom with fixed partnerships, the students of IES1 presented results in quantitative higher than the participants of IES2. Regarding knowledge about STI, although young people have reported that they do not know enough, they recognize some method for the prevention of infections. Participant sexual partnerships do not usually use the female condom. Alcohol and / or drug use was reported by most young people, but their employment did not precede their last sexual intercourse. In both institutions, much of it does not use the public health care system, has never tested for human immunodeficiency virus (HIV) and is not circumcised. It is concluded that the participants of the study assume a risk behavior and are vulnerable to STI and, in this context, educational actions that value the self-care of young people and guidelines related to the prevention of sexual health problems are opportune. It is necessary that the professionals involved in the care of the young population be trained and stimulate the reflection on the importance of the sexual health care, with emphasis on the male audience that does not frequently seek health services, adherence to prevention practices and risk behavior.

An observational clinical case of Zika virus-associated neurological disease is associated with primary IgG response and enhanced TNF levels.

Descriptive clinical data help to reveal factors that may provoke Zika virus (ZIKV) neuropathology. The case of a 24-year-old female with a ZIKV-associated severe acute neurological disorder was studied. The levels of ZIKV in the cerebrospinal fluid (CSF) were 50 times higher than the levels in other compartments. An acute anti-flavivirus IgG, together with enhanced TNF-alpha levels, may have contributed to ZIKV invasion in the CSF, whereas the unbiased genome sequencing [obtained by next-generation sequencing (NGS)] of the CSF revealed that no virus mutations were associated with the anatomic compartments (CSF, serum, saliva and urine).