Uncovering the neuroprotective effect of vitamin B12 in pneumococcal meningitis: insights into its pleiotropic mode of action at the transcriptional level.

Background: The interplay between bacterial virulence factors and the host innate immune response in pneumococcal meningitis (PM) can result in uncontrolled neuroinflammation, which is known to induce apoptotic death of progenitor cells and post-mitotic neurons in the hippocampal dentate gyrus, resulting in cognitive impairment. Vitamin B12 attenuates hippocampal damage and reduces the expression of some key inflammatory genes in PM, by acting as an epidrug that promotes DNA methylation, with increased production of S-adenosyl-methionine, the universal donor of methyl. Material and methods: Eleven-day-old rats were infected with S. pneumoniae via intracisternal injection and then administered either vitamin B12 or a placebo. After 24 hours of infection, the animals were euthanized, and apoptosis in the hippocampal dentate gyrus, microglia activation, and the inflammatory infiltrate were quantified in one brain hemisphere. The other hemisphere was used for RNA-Seq and RT-qPCR analysis. Results: In this study, adjuvant therapy with B12 was found to modulate the hippocampal transcriptional signature induced by PM in infant rats, mitigating the effects of the disease in canonical pathways related to the recognition of pathogens by immune cells, signaling via NF-kB, production of pro-inflammatory cytokines, migration of peripheral leukocytes into the central nervous system, and production of reactive species. Phenotypic analysis revealed that B12 effectively inhibited microglia activation in the hippocampus and reduced the inflammatory infiltrate in the central nervous system of the infected animals. These pleiotropic transcriptional effects of B12 that lead to neuroprotection are partly regulated by alterations in histone methylation markings. No adverse effects of B12 were predicted or observed, reinforcing the well-established safety profile of this epidrug. Conclusion: B12 effectively mitigates the impact of PM on pivotal neuroinflammatory pathways. This leads to reduced microglia activation and inflammatory infiltrate within the central nervous system, resulting in the attenuation of hippocampal damage. The anti-inflammatory and neuroprotective effects of B12 involve the modulation of histone markings in hippocampal neural cells.

Neurotoxic effects of hallucinogenic drugs 25H-NBOMe and 25H-NBOH in organotypic hippocampal cultures.

Introduction: NBOMes and NBOHs are psychoactive drugs derived from phenethylamines and have hallucinogenic effects due to their strong agonism to serotonin 5-HT2A receptors. Although cases of toxicity associated with the recreational use of substituted phenethylamines are frequently reported, there is a lack of information on the possible neurotoxic effects of NBOMe and NBOH in the brain hippocampus, a major neurogenesis region. Objectives: This study aimed at assessing the phenotypic and molecular effects of prolonged exposure of the hippocampus to the drugs 25H-NBOMe and 25H-NBOH. Methods: The ex vivo organotypic culture model of hippocampal slices (OHC) was used to investigate, by immunofluorescence and confocal microscopy, and transcriptome analyses, the mechanisms associated with the neurotoxicity of 25H-NBOMe and 25H-NBOH. Results: Reduction in the density of mature neurons in the OHCs occurred after two and seven days of exposure to 25H-NBOMe and 25H-NBOH, respectively. After the withdrawal of 25H-NBOMe, the density of mature neurons in the OHCs stabilized. In contrast, up to seven days after 25H-NBOH removal from the culture medium, progressive neuron loss was still observed in the OHCs. Interestingly, the exposure to 25H-NBOH induced progenitor cell differentiation, increasing the density of post-mitotic neurons in the OHCs. Corroborating these findings, the functional enrichment analysis of differentially expressed genes in the OHCs exposed to 25H-NBOH revealed the activation of WNT/Beta-catenin pathway components associated with neurogenesis. During and after the exposure to 25H-NBOMe or 25H-NBOH, gene expression patterns related to the activation of synaptic transmission and excitability of neurons were identified. Furthermore, activation of signaling pathways and biological processes related to addiction and oxidative stress and inhibition of the inflammatory response were observed after the period of drug exposure. Conclusion: 25H-NBOMe and 25H-NBOH disrupt the balance between neurogenesis and neuronal death in the hippocampus and, although chemically similar, have distinct neurotoxicity mechanisms.

Organotypic hippocampal culture model reveals differential responses to highly similar Zika virus isolates.

INTRODUCTION: Zika virus (ZIKV) caused an outbreak in Brazil, in 2015, being associated to microcephaly. ZIKV has a strong neurotropism leading to death of infected cells in different brain regions, including the hippocampus, a major site for neurogenesis. The neuronal populations of the brain are affected differently by ZIKV from Asian and African ancestral lineages. However, it remains to be investigated whether subtle variations in the ZIKV genome can impact hippocampus infection dynamics and host response. OBJECTIVE: This study evaluated how two Brazilian ZIKV isolates, PE243 and SPH2015, that differ in two specific missense amino acid substitutions, one in the NS1 protein and the other in the NS4A protein, affect the hippocampal phenotype and transcriptome. METHODS: Organotypic hippocampal cultures (OHC) from infant Wistar rats were infected with PE243 or SPH2015 and analyzed in time series using immunofluorescence, confocal microscopy, RNA-Seq and RT-qPCR. RESULTS: Unique patterns of infection and changes in neuronal density in the OHC were observed for PE243 and SPH2015 between 8 and 48 h post infection (p.i.). Phenotypic analysis of microglia indicated that SPH2015 has a greater capacity for immune evasion. Transcriptome analysis of OHC at 16 h p.i. disclosed 32 and 113 differentially expressed genes (DEGs) in response to infection with PE243 and SPH2015, respectively. Functional enrichment analysis suggested that infection with SPH2015 activates mostly astrocytes rather than microglia. PE243 downregulated biological process of proliferation of brain cells and upregulated those associated with neuron death, while SPH2015 downregulated processes related to neuronal development. Both isolates downregulated cognitive and behavioral development processes. Ten genes were similarly regulated by both isolates. They are putative biomarkers of early hippocampus response to ZIKV infection. At 5, 7, and 10 days p.i., neuronal density of infected OHC remained below controls, and mature neurons of infected OHC showed an increase in the epigenetic mark H3K4me3, which is associated to a transcriptionally active state. This feature is more prominent in response to SPH2015. CONCLUSION: Subtle genetic diversity of the ZIKV affects the dynamics of viral dissemination in the hippocampus and host response in the early stages of infection, which may lead to different long-term effects in neuronal population.

Riscos de contaminação por glúten em um restaurante universitário com preparações para indivíduos celíacos: um estudo de caso / Contamination risk for gluten in a restaurant university with preparations for individuals celiacs: a case study

Objetivo: Este estudo teve como objetivo avaliar os riscos de contaminação por glúten em um restaurante universitário com preparações para indivíduos celíacos. Métodos: Trata-se de um estudo observacional descritivo desenvolvido entre os meses de setembro e novembro de 2014 em um restaurante universitário do município de Belo Horizonte-MG, Brasil. Foi elaborado um check list baseado na literatura referente à doença celíaca e na Resolução n.º 275, de 21 de outubro de 2002, para identificar os riscos de contaminação cruzada por glúten de acordo com o fluxo de produção adotado pelo restaurante. A análise dos resultados foi realizada de forma qualitativa. Resultados e Discussão: Foram identificados oito principais pontos críticos que abrangem desde a recepção da matéria-prima à distribuição das refeições. Os dois pontos críticos identificados no check list como susceptíveis à altera- ção encontravam-se no almoxarifado. Os demais itens não apresentam possibilidade de alterações em curto prazo. Conclusões: Apesar dos manipuladores de alimentos do restaurante serem capacitados previamente para realizar preparações isentas de glúten, os riscos de contaminação por esta proteína são grandes. Nesse cenário faz-se necessá- rio um controle rigoroso em relação à linha de produção em um restaurante que distingue preparações com e sem glúten ou optar em realizar as preparações em outro espaço físico para garantir a inocuidade do alimento (AU)

Objective: This study aimed to evaluate the risks of cross-contamination of gluten in a university restaurant with preparations for celiac individuals. Methods: This is an observational descriptive study carried out between the months of September and November 2014 in a university restaurant in the city of Belo HorizonteMG, Brazil. A checklist was elaborated, based on literature related to celiac disease and on Resolution 275, issued on October 2002, to identify the risks of cross contamination by gluten according to the production flow adopted at the restaurant. The analysis was performed in a qualitative manner. Results and Discussion: Eight critical points were identified, ranging from the reception of raw material to the meal distribution at the restaurant’s dining hall. The two critical points identified in the checklist as susceptible to change were in the warehouse. The remaining items have no possibility of changes in the short term. Conclusions: Despite the restaurant food handlers have been trained previously to make preparations gluten-free, the risk of contamination by this protein are great. In this scenario it is necessary strict control over the production line in a restaurant that distinguishes preparations with and without gluten or choose to make preparations in another physical space to ensure the safety of food (AU)

Consumo de frutas e hortaliças e as condições de saúde de homens e mulheres atendidos na atenção primária à saúde / Consumption of fruit and vegetables and the health conditions of men and women attended in the primary healthcare network

Resumo Estudo transversal com o objetivo de analisar as associações entre as condições de saúde e a inadequação do consumo de frutas e hortaliças (FH) de 1.255 homens e mulheres atendidos em Unidades Básicas de Saúde de Belo Horizonte (MG). A coleta de dados contemplou o consumo de FH, a situação socioeconômica e as condições de saúde. Os resultados foram apresentados em razão de prevalência (RP) e intervalo de confiança de 95% (IC 95%). Verificou-se 77,5% (IC 95%: 75,1%-79,8%) de inadequação do consumo de FH (< 5 porções diárias) e esta foi superior entre os homens (83,8%, IC 95%: 79,0%-88,5% vs. mulheres: 76%, IC 95%: 73,4%-78,6%). Para as mulheres, o consumo inadequado de FH foi maior entre aquelas com percepção muito ruim da qualidade de sua saúde (RP: 1,37; IC 95%: 1,19-1,59) e entre as que consideraram a sua alimentação não saudável (RP: 1,15; IC 95%: 1,07-1,24). Para os homens, o consumo inadequado de FH foi superior entre os que referiram de 2 a 4 visitas ao médico no ano anterior à entrevista (RP: 1,21; IC 95%: 1,06-1,37). Concluiu-se que o consumo de FH na amostra encontra-se aquém das recomendações e que as condições de saúde se associaram de maneira distinta entre os sexos.

ABSTRACT This cross-sectional study sought to analyze the associations between health conditions and the inadequate consumption of fruit and vegetables (FV) of 1,255 men and women attended in Primary Healthcare Units in Belo Horizonte (State of Minas Gerais). Data collection included FV consumption, socioeconomic status and health conditions. Results are presented by prevalence ratio (PR) with a confidence interval of 95% (CI 95%). A 77.5% (CI 95%: 75.1%-79.8%) of inadequacy of consumption (< 5 daily servings) was found and it was more prevalent among men (83.8%, CI 95%: 79.0%-88.5% than among women: 76.0%, CI 95%: 73.4%-78.6%). For women, the inadequate consumption of FV was higher among those with poor perception of health quality (PR: 1.37; CI 95%: 1.19-1.59) and among those who consider their dietary habits as being unhealthy (PR: 1.15; CI 95%: 1.07-1.24). For men, the inadequate consumption was higher between individuals that reported 2 to 4 visits to the doctor in the year prior to the interview (PR: 1.21; CI 95%: 1.06-1.21). The conclusions showed that the consumption of FV among the population under study is below the recommendations and the health conditions are associated differently for each gender.

[Consumption of fruit and vegetables and the health conditions of men and women attended in the primary healthcare network]. / Consumo de frutas e hortaliças e as condições de saúde de homens e mulheres atendidos na atenção primária à saúde.

This cross-sectional study sought to analyze the associations between health conditions and the inadequate consumption of fruit and vegetables (FV) of 1,255 men and women attended in Primary Healthcare Units in Belo Horizonte (State of Minas Gerais). Data collection included FV consumption, socioeconomic status and health conditions. Results are presented by prevalence ratio (PR) with a confidence interval of 95% (CI 95%). A 77.5% (CI 95%: 75.1%-79.8%) of inadequacy of consumption (< 5 daily servings) was found and it was more prevalent among men (83.8%, CI 95%: 79.0%-88.5% than among women: 76.0%, CI 95%: 73.4%-78.6%). For women, the inadequate consumption of FV was higher among those with poor perception of health quality (PR: 1.37; CI 95%: 1.19-1.59) and among those who consider their dietary habits as being unhealthy (PR: 1.15; CI 95%: 1.07-1.24). For men, the inadequate consumption was higher between individuals that reported 2 to 4 visits to the doctor in the year prior to the interview (PR: 1.21; CI 95%: 1.06-1.21). The conclusions showed that the consumption of FV among the population under study is below the recommendations and the health conditions are associated differently for each gender.