Iron and vitamin D intake on a diet are able to modify the in vitro immune response to Mycobacterium leprae.

BACKGROUND: The impact of nutrient availability on the survival of Mycobacterium leprae and the development of leprosy remains largely unknown. Iron is essential for the survival and replication of pathogens, while vitamin D has been involved with pathogen elimination and immunoregulation. OBJECTIVES: We evaluated the influence of dietary iron and vitamin D supplementation and restriction on the inflammatory response of mouse immune cells in vitro. METHODS: After 30 days of standard or modified diets, peritoneal cells and splenocytes were stimulated with the alive microorganisms and sonicated antigens of M. leprae, respectively. The production of inflammatory cytokines, reactive oxygen species, and cell proliferation were evaluated. FINDINGS: In peritoneal cells, vitamin D supplementation and iron restriction reduced the production of IL-6 and TNF in response to M. leprae, while splenocytes presented a reduction in TNF production under the same conditions. Lower levels of IFN-γ and TNF were observed in both iron-supplemented and iron-deficient splenocytes. Besides, iron supplementation also reduced the production of IL-6 and IL-10. No changes in the production of reactive oxygen species or in cell proliferation were observed related to different diets. MAIN CONCLUSIONS: Taken together, these data point to an interference of the status of these nutrients on the interaction between the host and M. leprae, with the potential to interfere with the progression of leprosy. Our results highlight the impact of nutritional aspects on this neglected disease, which is significantly associated with unfavourable social conditions.

Iron and vitamin D intake on a diet are able to modify the in vitro immune response to Mycobacterium leprae

BACKGROUND The impact of nutrient availability on the survival of Mycobacterium leprae and the development of leprosy remains largely unknown. Iron is essential for the survival and replication of pathogens, while vitamin D has been involved with pathogen elimination and immunoregulation. OBJECTIVES We evaluated the influence of dietary iron and vitamin D supplementation and restriction on the inflammatory response of mouse immune cells in vitro. METHODS After 30 days of standard or modified diets, peritoneal cells and splenocytes were stimulated with the alive microorganisms and sonicated antigens of M. leprae, respectively. The production of inflammatory cytokines, reactive oxygen species, and cell proliferation were evaluated. FINDINGS In peritoneal cells, vitamin D supplementation and iron restriction reduced the production of IL-6 and TNF in response to M. leprae, while splenocytes presented a reduction in TNF production under the same conditions. Lower levels of IFN-γ and TNF were observed in both iron-supplemented and iron-deficient splenocytes. Besides, iron supplementation also reduced the production of IL-6 and IL-10. No changes in the production of reactive oxygen species or in cell proliferation were observed related to different diets. MAIN CONCLUSIONS Taken together, these data point to an interference of the status of these nutrients on the interaction between the host and M. leprae, with the potential to interfere with the progression of leprosy. Our results highlight the impact of nutritional aspects on this neglected disease, which is significantly associated with unfavourable social conditions.

Time between symptom and testing in relation to familial transmission of severe acute respiratory syndrome coronavirus 2 / Tempo entre o início dos sintomas e a testagem está relacionado à maior transmissão de SARS-CoV-2 entre contatos familiares

Abstract Brazil has a huge number of cases and deaths due to coronavirus disease 2019 (COVID-19); however, few studies have dealt with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection among familial contacts in Brazil. Here, we report our findings on transmission in a family-based study in Bauru, São Paulo, Brazil. The study, conducted from July to November 2020, comprised 974 individuals with 233 index patients and 741 familial contacts. Familial contacts were evaluated using the rapid COVID-19 Ag ECO and reverse transcription-polymerase chain reaction (RT-PCR) tests immediately after the index patient diagnosis. The antigen-based rapid test was validated in 121 individuals using RT-PCR as the gold standard. Additionally, 30 days later, familial contacts were evaluated for IgM and IgG antibodies against SARS-CoV-2. We found 333 cases of COVID-19 among familial contacts (44.9%). A positive correlation was observed between the time elapsed from the onset of symptoms until the index patient's COVID-19 testing and the number of family contacts infected by SARS-CoV-2. Early SARS-CoV-2 testing and familial contact evaluation are relevant strategies to contain transmission.

Resumo O Brasil apresenta um alto número de casos e óbitos por coronavírus (COVID-19), apesar disso, poucos estudos tratavam da infecção pelo coronavirus-2 causador de síndrome respiratória aguda grave (SARS-CoV-2) entre contatos familiares no Brasil. Relatamos aqui nossos achados sobre a transmissão de SARS-CoV-2 em um estudo de base familiar de Bauru, no estado de São Paulo, Brasil. O estudo foi realizado de julho a novembro de 2020 e compreendeu 974 indivíduos, sendo 233 pacientes índice e 741 contatos familiares. Os contatos familiares foram avaliados por meio do teste rápido COVID-19 Ag ECO Test e RT-PCR imediatamente após o diagnóstico do paciente índice. O uso do teste rápido baseado em antígeno foi validado em 121 indivíduos utilizando RT-PCR como padrão ouro. Adicionalmente, 30 dias após a avaliação inicial, os contatos familiares foram avaliados quanto à presença de anticorpos IgM e IgG contra SARS-CoV-2. Encontramos 333 casos de COVID-19 entre contatos familiares (44,9%). Observamos uma correlação positiva entre o tempo decorrido entre o início dos sintomas e o teste para COVID-19 do paciente índice e o número de contatos familiares infectados por SARS-CoV-2. A testagem precoce da infecção por SARS-CoV-2 e a avaliação de contatos familiares são estratégias relevantes para conter a transmissão.

Time between symptom and testing in relation to familial transmission of severe acute respiratory syndrome coronavirus 2.

Brazil has a huge number of cases and deaths due to coronavirus disease 2019 (COVID-19); however, few studies have dealt with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection among familial contacts in Brazil. Here, we report our findings on transmission in a family-based study in Bauru, São Paulo, Brazil. The study, conducted from July to November 2020, comprised 974 individuals with 233 index patients and 741 familial contacts. Familial contacts were evaluated using the rapid COVID-19 Ag ECO and reverse transcription-polymerase chain reaction (RT-PCR) tests immediately after the index patient diagnosis. The antigen-based rapid test was validated in 121 individuals using RT-PCR as the gold standard. Additionally, 30 days later, familial contacts were evaluated for IgM and IgG antibodies against SARS-CoV-2. We found 333 cases of COVID-19 among familial contacts (44.9%). A positive correlation was observed between the time elapsed from the onset of symptoms until the index patient's COVID-19 testing and the number of family contacts infected by SARS-CoV-2. Early SARS-CoV-2 testing and familial contact evaluation are relevant strategies to contain transmission.

Large-Scale Gene Expression Signatures Reveal a Microbicidal Pattern of Activation in Mycobacterium leprae-Infected Monocyte-Derived Macrophages With Low Multiplicity of Infection.

Leprosy is a disease with a clinical spectrum of presentations that is also manifested in diverse histological features. At one pole, lepromatous lesions (L-pole) have phagocytic foamy macrophages heavily parasitized with freely multiplying intracellular Mycobacterium leprae. At the other pole, the presence of epithelioid giant cells and granulomatous formation in tuberculoid lesions (T-pole) lead to the control of M. leprae replication and the containment of its spread. The mechanism that triggers this polarization is unknown, but macrophages are central in this process. Over the past few years, leprosy has been studied using large scale techniques to shed light on the basic pathways that, upon infection, rewire the host cellular metabolism and gene expression. M. leprae is particularly peculiar as it invades Schwann cells in the nerves, reprogramming their gene expression leading to a stem-like cell phenotype. This modulatory behavior exerted by M. leprae is also observed in skin macrophages. Here, we used live M. leprae to infect (10:1 multiplicity of infection) monocyte-derived macrophages (MDMs) for 48 h and analyzed the whole gene expression profile using microarrays. In this model, we observe an intense upregulation of genes consistent with a cellular immune response, with enriched pathways including peptide and protein secretion, leukocyte activation, inflammation, and cellular divalent inorganic cation homeostasis. Among the most differentially expressed genes (DEGs) are CCL5/RANTES and CYP27B1, and several members of the metallothionein and metalloproteinase families. This is consistent with a proinflammatory state that would resemble macrophage rewiring toward granulomatous formation observed at the T-pole. Furthermore, a comparison with a dataset retrieved from the Gene Expression Omnibus of M. leprae-infected Schwann cells (MOI 100:1) showed that the patterns among the DEGs are highly distinct, as the Schwann cells under these conditions had a scavenging and phagocytic gene profile similar to M2-like macrophages, with enriched pathways rearrangements in the cytoskeleton, lipid and cholesterol metabolism and upregulated genes including MVK, MSMO1, and LACC1/FAMIN. In summary, macrophages may have a central role in defining the paradigmatic cellular (T-pole) vs. humoral (L-pole) responses and it is likely that the multiplicity of infection and genetic polymorphisms in key genes are gearing this polarization.

Large-scale gene expression signatures reveal a microbicidal pattern of activation in Mycobacterium leprae-infected monocyte-derived macrophages with low multiplicity of infection

Leprosy is a disease with a clinical spectrum of presentations that is also manifested in diverse histological features. At one pole, lepromatous lesions (L-pole) have phagocytic foamy macrophages heavily parasitized with freely multiplying intracellular Mycobacterium leprae. At the other pole, the presence of epithelioid giant cells and granulomatous formation in tuberculoid lesions (T-pole) lead to the control of M. leprae replication and the containment of its spread. The mechanism that triggers this polarization is unknown, but macrophages are central in this process. Over the past few years, leprosy has been studied using large scale techniques to shed light on the basic pathways that, upon infection, rewire the host cellular metabolism and gene expression. M. leprae is particularly peculiar as it invades Schwann cells in the nerves, reprogramming their gene expression leading to a stem-like cell phenotype. This modulatory behavior exerted by M. leprae is also observed in skin macrophages. Here, we used live M. leprae to infect (10:1 multiplicity of infection) monocyte-derived macrophages (MDMs) for 48 h and analyzed the whole gene expression profile using microarrays. In this model, we observe an intense upregulation of genes consistent with a cellular immune response, with enriched pathways including peptide and protein secretion, leukocyte activation, inflammation, and cellular divalent inorganic cation homeostasis. Among the most differentially expressed genes (DEGs) are CCL5/RANTES and CYP27B1, and several members of the metallothionein and metalloproteinase families. This is consistent with a proinflammatory state that would resemble macrophage rewiring toward granulomatous formation observed at the T-pole. Furthermore, a comparison with a dataset retrieved from the Gene Expression Omnibus of M. leprae-infected Schwann cells (MOI 100:1) showed that the patterns among the DEGs are highly distinct, as the Schwann cells under these conditions had a scavenging and phagocytic gene profile similar to M2-like macrophages, with enriched pathways rearrangements in the cytoskeleton, lipid and cholesterol metabolism and upregulated genes including MVK, MSMO1, and LACC1/FAMIN. In summary, macrophages may have a central role in defining the paradigmatic cellular (T-pole) vs. humoral (L-pole) responses and it is likely that the multiplicity of infection and genetic polymorphisms in key genes are gearing this polarization.

Reuse of wastewater from pulp industry for the optimization of fungal xylanase production / Reutilização de efluentes da indústria de celulose para otimização da produção de xilanase fúngica

The production of enzymes using agro-industrial waste is a low cost alternative for the reuse of byproducts, with the subsequent impact decrease on the environment. Current analysis produced xylanase using fungus Aspergillus niger, with two types of wastewater generated during the pulp chemical bleaching phase as inducers. Xylanase was produced by submerged liquid fermentation and factorial design optimized parameters that influence production (concentration of wastewater and production period). Initial culture conditions (pH, temperature and agitation) were optimized independently. Alkaline wastewater was more effective than acidic wastewater for the induction of xylanase in optimized conditions: 50% of culture medium, 7-day production, 30C, pH 6.0 and agitation at 160 rpm. Despite different results, acidic and alkaline wastewaters induced xylanase production by A. niger when employed in concentrations lower than or equal to 50% of culture medium and in the most optimal conditions described above. Alkaline wastewater is highlighted as the most efficient for such production. 

A produção de enzimas, a partir de resíduos agroindustriais, é uma alternativa para reutilização destes subprodutos como substrato de baixo custo reduzindo seu impacto causado pelo descarte no meio ambiente. Diante disso, o objetivo deste estudo foi a produção de xilanase por Aspergillus niger, utilizando dois efluentes gerados nas fases de branqueamento químico de polpa de celulose como indutores. A produção de xilanase foi realizada por fermentação líquida submersa, e utilizou-se planejamento fatorial para otimização dos parâmetros influentes de produção (concentração de efluentes e período de produção) e as condições iniciais de cultivo (pH, temperatura e agitação) foram otimizadas de forma independente. O efluente alcalino se mostrou mais efetivo do que o efluente com característica ácida, na indução de xilanase em condições otimizadas: 50% em relação ao meio de cultura, sete dias de produção, 30°C, pH 6,0 e agitação de 160 rpm. Conclui-se que ambos os efluentes, ácido e alcalino, apesar de levarem a diferentes resultados, são capazes de induzir a produção de xilanase por A. niger quando empregues em concentrações menores ou iguais a 50% em relação ao meio de cultura e nas condições ótimas descritas acima, destacando-se o efluente alcalino como mais eficiente para produção de tal enzima.(AU)

Reuse of wastewater from pulp industry for the optimization of fungal xylanase production / Reutilização de efluentes da indústria de celulose para otimização da produção de xilanase fúngica

The production of enzymes using agro-industrial waste is a low cost alternative for the reuse of byproducts, with the subsequent impact decrease on the environment. Current analysis produced xylanase using fungus Aspergillus niger, with two types of wastewater generated during the pulp chemical bleaching phase as inducers. Xylanase was produced by submerged liquid fermentation and factorial design optimized parameters that influence production (concentration of wastewater and production period). Initial culture conditions (pH, temperature and agitation) were optimized independently. Alkaline wastewater was more effective than acidic wastewater for the induction of xylanase in optimized conditions: 50% of culture medium, 7-day production, 30°C, pH 6.0 and agitation at 160 rpm. Despite different results, acidic and alkaline wastewaters induced xylanase production by A. niger when employed in concentrations lower than or equal to 50% of culture medium and in the most optimal conditions described above. Alkaline wastewater is highlighted as the most efficient for such production.

A produção de enzimas, a partir de resíduos agroindustriais, é uma alternativa para reutilização destes subprodutos como substrato de baixo custo reduzindo seu impacto causado pelo descarte no meio ambiente. Diante disso, o objetivo deste estudo foi a produção de xilanase por Aspergillus niger, utilizando dois efluentes gerados nas fases de branqueamento químico de polpa de celulose como indutores. A produção de xilanase foi realizada por fermentação líquida submersa, e utilizou-se planejamento fatorial para otimização dos parâmetros influentes de produção (concentração de efluentes e período de produção) e as condições iniciais de cultivo (pH, temperatura e agitação) foram otimizadas de forma independente. O efluente alcalino se mostrou mais efetivo do que o efluente com característica ácida, na indução de xilanase em condições otimizadas: 50% em relação ao meio de cultura, sete dias de produção, 30°C, pH 6,0 e agitação de 160 rpm. Conclui-se que ambos os efluentes, ácido e alcalino, apesar de levarem a diferentes resultados, são capazes de induzir a produção de xilanase por A. niger quando empregues em concentrações menores ou iguais a 50% em relação ao meio de cultura e nas condições ótimas descritas acima, destacando-se o efluente alcalino como mais eficiente para produção de tal enzima.

Prevalência de Streptococcus agalactiae em secreção vaginal de gestantes atendidas em um laboratório de análises clínicas do interior do Estado de São Paulo / Prevalence of Streptococcus agalactiae in vaginal secretion of pregnant women attended at a laboratory of the interior State of São Paulo

Introdução: os agentes responsáveis por infecções graves em neonatais são geralmente oriundos da mãe, tendo como o grupo mais grave e predominante os Streptococcus do grupo B (EGB). Estas bactérias Gram-positivas, normalmente presentes no trato gastrointestinal, podem ser transmitidas da mãe para o feto através de transmissão vertical, gerando graves doenças em neonatos, com taxas de mor talidade de 50% e de colonização de aproximadamente 6 a 8% das gestantes. Objetivos: este estudo avaliou a prevalência de gestantes portadoras de Streptococcus agalactiae, atendidas no Laboratório de Análises Clínicas da Fundação Véritas da Universidade do Sagrado Coração no município de Bauru no período de 2013 a 2015. Material e Métodos: foram analisados laudos de exames de cultura de secreção vaginal de pacientes gestantes disponíveis no sistema Pleres, utilizando um formulário para coleta das informações. Resultados: os resultados demonstraram que no período de 2013 a 2015, das 560 culturas de secreção vaginal realizadas para pacientes gestantes cerca de 4,3% apresentaram-se positivas para S. agalactiae, sendo este percentual composto em sua maioria por atendimentos originários do Sistema Único de Saúde (SUS), além da prevalência de gestantes entre 20 e 29 anos. Dentre os antimicrobianos avaliados nos testes de suscetibilidade observou-se que S. agalactiae foi resistente, especialmente, à clindamicina. Conclusão: foram encontradas taxas de prevalência de S. agalactiae em gestantes menores que as médias observadas em outros estudos nacionais, com perfil de sensibilidade reduzido frente à clindamicina.

Introduction: the agents responsible for the serious infections in newborns are generally coming in the mother, having the Group B Streptococcus (GBS) as the most severe and prevalent group. These agents are Gram-positive bacteria that are normally found in the gastrointestinal tract and can be transmitted from the mother to the unborn baby by vertical transmission, which generates many severe diseases in mothers who have just given birth and newborn babies, with mortality rates at 50%, these bacteria colonize approximately 6 to 8% of pregnant women. Objectives: this epidemiological study aimed to evaluate the prevalence of pregnant women bearing Streptococcus agalactiae (EGB) in their birth canal and who were taken care of at the Fundação Véritas Clinical Analysis Laboratory of Universidade do Sagrado Coração in the city of Bauru, during the period 2013/2015. Material and methods: we analyzed exam reports of vaginal secretion culture tests for GBS of pregnant patients available in the Pleres system, using a form for collecting the information. Results and Discussion: the results showed that of 560 cultures performed during this period, 4.3% had S. agalactiae isolated, being that most of the patients were admitted by the Brazilian Sistema Único de Saúde (SUS) and had ages ranging from 20 to 29 years old. Among the antibiotics evaluated in the sensitivity tests, S. agalactiae proved to be resistant, especially, to clindamycin. Conclusion: it is believed that the decrease in the percentage of prevalence of S. agalactiae in this study in comparison to other researches was due to the changes in public policies and the implementation of screening protocols from 2013 for ANVISA (Agência Nacional de Vigilância Sanitária).