Determining reference ranges for immunological cells of healthy indigenous individuals from a region in Brazil.

Morais et al. conducted a pioneering study with Brazilian indigenous populations to determine reference values for immunologic cells from healthy adult individuals. The main findings included a higher relative median for T lymphocyte subsets in females than males, and T CD3+, T CD4+, and T CD8+ relative values were statistically different when compared with Brazilian populations from other Brazilian regions. The relative medians of CD3+, CD4+, and CD8+ T cells were significantly higher in women than in men in a healthy indigenous population. Demographic and ethnic diversity of the Brazilian population can be associated with quantitative modifications in the immunologic cells of healthy individuals. OBJECTIVE: The establishment of reference values for a subset of leukocytes is common in clinical practice, and ethnic variations are strongly associated with disease development. In Brazil, indigenous people are vulnerable to infections, and few studies have described the health and disease conditions of this population. This study aimed to provide reference values for immunological cell subsets in indigenous Brazilians living in the state of Mato Grosso do Sul. METHODS: Flow cytometry and 4-color combinations of monoclonal antibodies were used to characterize cells. A total of 115 healthy adults, mostly females (72%), were included in the study. The results are presented as mean and median (2.5%-97.5% percentiles) for T and B lymphocytes, CD4+ T cells, CD8+ T cells, Natural Killer cells, monocytes, and dendritic cells, providing an average immunological profile for the population in question. RESULTS: The relative medians of CD3+, CD4+, and CD8+ T cells were significantly higher in women than in men in a healthy indigenous population. CONCLUSION: To our knowledge, cell reference data from indigenous Brazilians are unknown in the literature. The immune cell results presented in this pioneering study will contribute to the clinical and laboratory evaluation of the Brazilian indigenous population, especially given the important differences when compared with other Brazilian ethnic groups.

Systemic Inflammatory Molecules Are Associated with Advanced Fibrosis in Patients from Brazil Infected with Hepatitis Delta Virus Genotype 3 (HDV-3).

BACKGROUND AND AIMS: Hepatitis Delta virus (HDV) genotype 3 is responsible for outbreaks of fulminant hepatitis in Northeastern South America. This study investigates if systemic inflammatory molecules are differentially expressed in patients with advanced fibrosis chronically infected with Hepatitis Delta virusgenotype 3(HDV-3). METHODS: Sixty-one patients from the north of Brazil coinfected with hepatitis B virus (HBV)/HDV-3 were analyzed. HDV quantification and genotyping were performed by semi-nested real-time polymerase chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP) methodologies. Ninety-two systemic inflammatory molecules (SIMs) were measured by Proximity Extension Assay (PEA) technology. The Shapiro-Wilk, Student's t-test, Mann-Whitney tests, and logistic regression analysis were used when appropriate. RESULTS: The median age was 41 years, and all patients were HBeAg negative. Advanced fibrosis or cirrhosis was diagnosed by histological staging in 17 patients, while 44 presented with minimal or no fibrosis. Advanced necroinflammatory activity correlated positively with serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT). Established non-invasive fibrosis scores (APRI, FIB-4, and AST/ALT ratio) revealed low sensitivities and positive predictive values (PPVs) with an AUROC maximum of 0.586. Among the 92 SIMs analyzed, MCP.4, CCL19, EN.RAGE, SCF, and IL18 showed a positive correlation with fibrosis stage. A combined score including CCL19 and MCP.4 revealed a sensitivity of 81% and an odds ratio of 2.202 for advanced fibrosis. CONCLUSIONS: Standard non-invasive fibrosis scores showed poor performance in HDV-3 infection. We here suggest that the determination of CCL19 and MCP.4 may be used to identify patients with advanced fibrosis. Moreover, this study gives novel insights into the immunopathogenesis of HDV-3 infection.

Determining reference ranges for immunological cells of healthy indigenous individuals from a region in Brazil

ABSTRACT Objective The establishment of reference values for a subset of leukocytes is common in clinical practice, and ethnic variations are strongly associated with disease development. In Brazil, indigenous people are vulnerable to infections, and few studies have described the health and disease conditions of this population. This study aimed to provide reference values for immunological cell subsets in indigenous Brazilians living in the state of Mato Grosso do Sul. Methods Flow cytometry and 4-color combinations of monoclonal antibodies were used to characterize cells. A total of 115 healthy adults, mostly females (72%), were included in the study. The results are presented as mean and median (2.5%-97.5% percentiles) for T and B lymphocytes, CD4+ T cells, CD8+ T cells, Natural Killer cells, monocytes, and dendritic cells, providing an average immunological profile for the population in question. Results The relative medians of CD3+, CD4+, and CD8+ T cells were significantly higher in women than in men in a healthy indigenous population. Conclusion To our knowledge, cell reference data from indigenous Brazilians are unknown in the literature. The immune cell results presented in this pioneering study will contribute to the clinical and laboratory evaluation of the Brazilian indigenous population, especially given the important differences when compared with other Brazilian ethnic groups.

The Association of Bacterin and Recombinant Proteins Induces a Humoral Response in Sheep against Caseous Lymphadenitis.

In this study, we investigated the capacity of the recombinant proteins SpaC, NanH, SodC, and PLD of C. pseudotuberculosis to trigger protective humoral and cellular immune responses against experimentally induced C. pseudotuberculosis infection in sheep. The antigens were produced in a heterologous system and were purified by affinity chromatography. Nine sheep were randomly divided into three groups, which were immunized as follows: Group 1 (control)-a mix of adjuvants composed of the inactivated T1 strain of C. pseudotuberculosis and commercial Montanide™ISA 61 VG (T1M); Group 2-rSpaC, rSodC, rPLD, and T1M; Group 3-rNanH, rSodC, rPLD, and T1M. All groups were immunized twice (on days 0 and 30) and challenged on day 90 of the experiment. Humoral and cellular immune responses were evaluated by Enzyme-Linked Immunosorbent Assay (ELISA) to quantify the IgG antibodies and interferon-gamma (IFN-y). Both vaccine formulations with recombinant proteins (groups 2 and 3) could induce a significant humoral IgG immune response in sheep. The proteins rSodC, rPLD, and rNanH were more immunogenic, inducing significant levels of IgG antibodies after the first dose of the vaccine or after the challenge, maintaining constant levels until the end of the experiment. However, it was not possible to differentiate between the cellular responses induced by the vaccines. This lack of effectiveness points toward the need for further studies to improve the efficacy of this subunit-based vaccine approach.

Assessment of rSodC, rPknG, rNanH, and rSpaC as Antigens for Diagnostic Tools Against Caseous Lymphadenitis.

Corynebacterium pseudotuberculosis is a bacillus that causes caseous lymphadenitis in small ruminants, leading to great losses to rural producers; thus, an efficient diagnosis is necessary for using disease control measures. This study aimed to evaluate the antigenic potential of four C. pseudotuberculosis recombinant proteins (rSodC, rPknG, rNanH, and rSpaC) against sera of goat and sheep experimentally infected with one of three different C. pseudotuberculosis strains. Goats were infected with CAP76 or CAP21 strain (n = 10), sheep with VD57 strain (n = 6), and a group of not-infected animals (goats and sheep) were kept as a healthy control (healthy n = 12). Sera were collected at 0, 14, 60, 90, 180, or 190 days after inoculation for antigenicity testing using Western blotting and enzyme-linked immunosorbent assay (ELISA) techniques. Cross-reactivity tests with recombinant proteins were performed in goat serum experimentally vaccinated with Nocardia sp. or Rhodococcus equi bacterin. The rSodC protein showed discriminatory antigenic reactivity with a statistically significant difference against three different C. pseudotuberculosis strains evaluated in goats and sheep samples, while rPknG showed statistical significance only against two C. pseudotuberculosis strains evaluated in goats. rSodC was proved to be a strong candidate as a tool for diagnosis of C. pseudotuberculosis infection, once it was able to recognize antibodies against all strains evaluated in goats and sheep.

Severe Cases of COVID-19 and High Association with Causes of Immune Dysregulation: A Systematic Review.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the virus responsible for coronavirus 2019 (COVID-19), which was declared a pandemic in March 2020 by the World Health Organization due the rapid spread representing a global health crisis. The disease is characterized by a wide clinical spectrum ranging from asymptomatic forms until severe viral pneumonia, which can to evolve to severe acute respiratory syndrome, especially in elderly patients and/or with comorbidities. An efficient assembly of the immunological response of the patients becomes fundamental against SARS-CoV-2 infection and it has been demonstrating a significant relationship between the severity of the disease and expression profile of the immune cells and the levels of pro-inflammatory cytokines. This review aims to presents the main immunological mechanisms developed during the infection by SARS-CoV-2 in the evolution of the severe cases of COVID-19. The immune dysregulation of the Th1 cellular response standard, the instability in the production of neutralizing antibodies by plasma B cells, the difference in tropism of CD8+ T cells against virus proteins in early infection, late infection and reinfections, dynamic of alveolar macrophages and pulmonary innate lymphoid cells (TCR γδ) of the natural imune response and the high level of pro-inflammatory cytokines can determine the main cause of breath tissues damages and, consequently, a greater severity of the disease. Therefore, a complete understanding of the main immunological changes involved in SARS-CoV-2 infection can identify possible biomarkers in the evaluation of early prognosis of the severe cases of COVID-19, making possible better therapeutic success to the patients.

Quantification and Comprehensive Analysis of Mesenchymal Stromal Cells in Bone Marrow Samples from Sickle Cell Disease Patients with Osteonecrosis.

The potential use of bone marrow mesenchymal stromal cells (BM-MSCs) for the treatment of osteonecrosis in sickle cell disease (SCD) patients is increasing. However, convenient BM-MSC quantification and functional property assays are critical factors for cell-based therapies yet to be optimized. This study was designed to quantify the MSC population in bone marrow (BM) samples from SCD patients with osteonecrosis (SCD group) and patients with osteoarticular complications not related to SCD (NS group), using flow cytometry for CD271+CD45-/low cell phenotype and CFU-F assay. We also compared expanded BM-MSC osteogenic differentiation, migration, and cytokine secretion potential between these groups. The mean total cell number, CFU-F count, and CD271+CD45-/low cells in BM mononuclear concentrate were significantly higher in SCD than in NS patients. A significant correlation between CD271+CD45-/low cell number and CFU-F counts was found in SCD (r = 0.7483; p = 0.0070) and NS (r = 0.7167; p = 0.0370) BM concentrates. An age-related quantitative reduction of CFU-F counts and CD271+CD45-/low cell number was noted. Furthermore, no significant differences in the morphology, replicative capacity, expression of surface markers, multidifferentiation potential, and secretion of cytokines were found in expanded BM-MSCs from SCD and NS groups after in vitro culturing. Collectively, this work provides important data for the suitable measurement and expansion of BM-MSC in support to advanced cell-based therapies for SCD patients with osteonecrosis.

Frequency and factors associated with Toxoplasma gondii infection in pregnant women and their pets in Ilhéus, Bahia, Brazil.

INTRODUCTION: Toxoplasmosis is an asymptomatic disease that can lead to systemic disease in the fetus of pregnant women with primary infection. This study aimed to determine the prevalence of toxoplasmosis, associated factors, and correlation between the serology of pregnant women and their pets, in the municipality of Ilhéus, Bahia, Brazil. METHODS: This cross-sectional study was conducted in 196 pregnant women and their cats or dogs (n=89). Semi-structured interviews were conducted and serum samples from the pregnant women were tested to detect IgM and IgG antibodies against Toxoplasma gondii, and avidity tests were performed for IgM-positive samples. The serum collected from pets were tested for IgG antibodies, and IgM antibodies in cats. A non-conditional logistic regression analysis was performed to identify infection-associated factors. RESULTS: IgG and IgM antibodies were detected in 67.9% (133/196) and 1.5% (3/196) samples, respectively, for women with an avidity of over 60%. Age ≥ 25 and the presence of cats in the vicinity were found to be associated with infection, while the level of education and previous orientation toward prevention of toxoplasmosis were protective factors in pregnant women. IgG antibodies were detected in 46.1% (41/89) of the animals, and cats were found to be negative for IgM. For the animals, age ≥ 1 year was a factor associated with infection. There was no correlation between serology of the pregnant women and the animals (p=0.15). CONCLUSIONS: An elevated prevalence of toxoplasmosis was detected in the region. Therefore, the adoption of preventive measures by public healthcare bodies is recommended.

Seroepidemiology of hepatitis B in individuals born between 1945-1985 on a Brazilian regional metropolis / Soroepidemiologia da hepatite B em indivíduos nascidos entre 1945-1985 em uma metrópole regional Brasileira

Background: hepatitis B prevalence can be influenced by social/cultural behavior and the Baby Boomer (BB) generation(1945-1964) may have been more susceptible to this infection. Objectives: We investigated the seroprevalence of markers for HBV infection and vaccination and its association with main risk factors. Methodology: a random sample of individuals aged 30-70 years old in a public clinical laboratory from a metropolitan area of Bahia/Brazil were tested for HBsAg/Total Anti-HBc/Anti-HBs/Anti-HBc-IgM and a socio-demographic questionnaire was applied. Results: of the650 participants, 349 were 51-70 yo (BB) and 301 were non-BB. The prevalences were HBsAg (2.3%), Total Anti-HBc (17.1%) and Anti-HBs (27.4%). Anti-HBcIgM (2.7%) was performed in 112 participants sera who had contact/infection with HBV. The laboratory profiles were characterized as susceptibility (68%), vaccine response (14.8%) and contact/infection with HBV (17.2%). BB participants were more susceptible and less vaccinated than non-BB. The higher frequency of contact/infection status was observed in the BB generation. Statistically significant differences were found for the contact/infection status in males(50,9%) illicit drug use (11,6%), syringe/needle sharing (7,1%), and blood transfusion (10,7%). Non-BB with contact/ infection profile reported more tattoo/piercing and BB reported higher use of glass syringes. Conclusion: the majority of the study population was susceptible to infection but participants older than 50 years showed both, a higher frequency of this profile and also a higher frequency of contact/infection status, thus suggesting the need for greater health care attention for this age group.

Introdução: a prevalência de hepatite B pode ser influenciada pelo comportamento sociocultural e a geração Baby Boomer (BB) (1945-1964) pode ter sido mais suscetível a esta infecção. Objetivos: Investigar a soroprevalência de marcadores para a infecção pelo VHB e resposta vacinal e associação com fatores de risco. Metodologia: soro de indivíduos de 30 a 70 anos randomicamente selecionados em um laboratório publico de análises clínicas na área metropolitana do Brasil foram testados para AgHBs/ Anti-HBc Total /Anti-HBs/Anti-HBc-IgM. Todos responderam questionário sociodemográfico contendo perguntas sobre fatores de risco para hepatite B. Resultados: dos 650 participantes, 349 eram BB (51-70 anos) e 301 eram não-BB (30-50 anos). As prevalências estimadas foram: HBsAg (2,3%), Anti-HBs (27,4%). Entre os Anti-HBc Total (17,1%) apenas 2,7% foram Anti-HBc IgM. Os perfis laboratoriais foram caracterizados como suscetibilidade (68%), resposta vacinal (14,8%) e contato com VHB (17,2%). Na distribuição por idade, os BB foram mais susceptíveis, menos vacinados e apresentaram maior frequência de contato/infecção que os não-BB. Diferenças estatisticamente significantes foram encontradas no status contato/infecção e as seguintes variáveis: sexo masculino, uso de drogas ilícitas, compartilhamento de seringas de vidro/agulhas e transfusão de sangue. Não-BB com status contato/infecção relataram ter mais tatuagem/piercing e BB relataram maior uso de seringas de vidro. Conclusão: a maioria da população estudada era suscetível ao VHB, mas os participantes com mais de 50 anos apresentaram tanto uma maior frequência desse status quanto do status contato/ infecção, sugerindo a necessidade de maior atenção à saúde para indivíduos desta faixa etária.

Frequency and factors associated with Toxoplasma gondii infection in pregnant women and their pets in Ilhéus, Bahia, Brazil

Abstract INTRODUCTION: Toxoplasmosis is an asymptomatic disease that can lead to systemic disease in the fetus of pregnant women with primary infection. This study aimed to determine the prevalence of toxoplasmosis, associated factors, and correlation between the serology of pregnant women and their pets, in the municipality of Ilhéus, Bahia, Brazil. METHODS: This cross-sectional study was conducted in 196 pregnant women and their cats or dogs (n=89). Semi-structured interviews were conducted and serum samples from the pregnant women were tested to detect IgM and IgG antibodies against Toxoplasma gondii, and avidity tests were performed for IgM-positive samples. The serum collected from pets were tested for IgG antibodies, and IgM antibodies in cats. A non-conditional logistic regression analysis was performed to identify infection-associated factors. RESULTS: IgG and IgM antibodies were detected in 67.9% (133/196) and 1.5% (3/196) samples, respectively, for women with an avidity of over 60%. Age ≥ 25 and the presence of cats in the vicinity were found to be associated with infection, while the level of education and previous orientation toward prevention of toxoplasmosis were protective factors in pregnant women. IgG antibodies were detected in 46.1% (41/89) of the animals, and cats were found to be negative for IgM. For the animals, age ≥ 1 year was a factor associated with infection. There was no correlation between serology of the pregnant women and the animals (p=0.15). CONCLUSIONS: An elevated prevalence of toxoplasmosis was detected in the region. Therefore, the adoption of preventive measures by public healthcare bodies is recommended.

Temas de Bioética e de Biossegurança para uso de professores da educação secundária na preparação da autonomia de jovens entre 12 e 17 anos / Bioética y Bioseguridad para uso de profesores de la educación secundaria en la promoción de la autonomía de jóvenes entre 12 y 17 años / Bioethics and biosafety issues for the use of teachers in secondary education in the preparation of the autonomy of young people between 12 and 17 years / Bioètica i bioseguretat per a ús de professors d'educació secundària en la promoció de l'autonomia de joves entre 12 i 17 anys

A educação é crucial para a alocação da liberdade de decisão e autonomia, que possibilita a real dignidade aos indivíduos. A compreensão dos avanços científico-tecnológicos favorece a redução das desigualdades sócio-político-econômicas e culturais na população. Este artigo propõe temas para a abordagem dos professores sobre bioética e biossegurança no ensino secundário. Os professores podem escolher e adaptar temas, de acordo com as indicações institucionais, sua experiência e percepção, considerando o tempo, as idades, as características regionais e a realidade dos alunos

En los últimos años, el consentimiento informado del paciente ha asumido un papel tan importante en la relación médico-paciente, que ha cambiado el sistema de responsabilidad penal del médico en caso de intervención sin consentimiento, incluso en casos con resultados exitosos. También ha habido propuestas legislativas para introducir el delito de intervención médica arbitraria, pareciendo muy difícil encontrar una solución a este conflicto. En la actualidad, el Parlamento italiano examina una propuesta de ley sobre voluntades anticipadas, centradas en el consentimiento del paciente, que pueden no prestarse, o revocarse después de haberse dado, también para tratamientos como la nutrición y la hidratación artificial, eximiendo de toda responsabilidad civil y penal al médico. Es evidente que tal énfasis en el consentimiento informado, que involucra con tal fuerza al paciente en su tratamiento, está tomando un camino peligroso que puede vaciar por completo la alianza terapéutica médico-paciente que se juega en el campo de la atención médica

In recent years, informed consent of the patient has assumed such an important role in the physician-patient relationship that it has changed the system of criminal liability of the physician in case of intervention without consent, even in cases with successful results. There have also been legislative proposals to introduce the crime of arbitrary medical intervention, making it difficult to find a solution to this conflict. At present, the Italian Parliament is examining a draft bill on advanced directives, focusing on patient consent, which may not be given, or may be revoked after they have been given, also for treatments such as artificial nutrition and hydration, exempting the doctor from all civil and criminal liability. It is evident that such an emphasis on informed consent, which involves the patient in his treatment, is taking a dangerous path that can completely empty the doctor-patient therapeutic alliance played out in the field of medical care

L'educació és fonamental per a la llibertat i l'autonomia, que donen dignitat a les persones. La comprensió dels avanços científics i tecnològics afavoreix la reducció de les desigualtats socioculturals i políticoeconòmiques entre els ciutadans d'una població. En aquest article es proposen exemples de temes de bioètica i de bioseguretat per a ús de docents a l'escola. Els mestres poden adaptar els temes transversalment segons les exigències institucionals, la seva experiència i percepció, considerant les edats, temps, característiques regionals i realitats dels seus alumnes

O receptor de quimiocina CXCR3/CD183 na tuberculose / The chemokine receptor CXCR3/CD183 in tuberculosis

Introdução: o receptor CXCR3/CD183 juntamente com seu indutor IFNy e seus ligantes CXCL9, CXCL10 e CXCL11 têm sido descritos como de grande importância na resposta imune do perfil T helper 1 (Th1). Este grupo de quimiocinas é expresso no microambiente e permite a migração de células ao sítio da infecção para combater o patógeno. Objetivo: revisar o atual estado da arte sobre o papel do receptor CXCR3/CD183 na tuberculose. Metodologia: o presente estudo inclui a revisão narrativa de 12 artigos que foram selecionados a partir de 74 artigos encontrados nas bases de dados PubMed e Sciencedirect entre primeiro de agosto e 31 de outubro de 2014. Resultados: diferentes abordagens vêm sendo utilizadas para o estudo desse receptor. A utilização de modelos animais como camundongos, coelhos e macacos é a mais comum. Porém, ensaios in vitro com células humanas do sangue periférico e efusão pleural também já foram utilizados para representar, com maior fidelidade, a resposta ao Mycobacterium tuberculosis (Mtb) pelo sistema imune humano. Esses estudos resultaram em importantes achados sobre o papel do receptor CXCR3 na tuberculose (TB), principalmente quanto à expressão em linfócitos e neutrófilos, assim como o padrão de coexpressão de outros receptores. Conclusão: o CXCR3 é o receptor de uma importante citocina (IP-10) induzida pelo IFN-gama, produzida na resposta Th1, eficaz na resposta à tuberculose. Nesse trabalho, resssalta-se que foram encontrados poucos estudos sobre o tema e isso demonstra a necessidade de realização de novas pesquisas, a fim de melhor investigar o papel desse importante receptor na tuberculose.

Introduction: the CXCR3/CD183 receptor along with its IFNy inducer and its ligands: the chemokines named CXCL9, CXCL10 and CXCL11 are of great importance in the Th1 (T helper 1) immune response. This group of chemokine modulates the migration of cells to the site of infection to defend against the pathogen. Objective: to investigate the current state of the art on the role of the receptor CXCR3/CD183 in tuberculosis. Methodology: the present study includes the narrative review of 12 articles that were selected from 74 articles found in the PubMed and Sciencedirect databases between August 1 and October 31, 2014. Results: different approaches have been used for the study of this receptor. The use of animal models such as mice, rabbits and monkeys is more common. However, in vitro assays with human peripheral blood cells and pleural effusion were also used to represent more faithfully the response to Mycobacterium tuberculosis (Mtb) by the human immune system. These studies resulted in significant findings on the role of the CXCR3 receptor in tuberculosis (TB), especially for expression in lymphocytes and neutrophils, as well as the pattern of co-expression of other receptors. Conclusion: CXCR3 is the receptor for an important cytokine (IP-10) induced by IFN-gamma, produced in the Th1 response, effective in responding to tuberculosis. In this work, it is emphasized that cheeses found few studies on the subject and demonstration, the need for conducting research, in order to better investigate the role of this important receptor in tuberculosis.

Interferon-gamma release assay performance in northeastern Brazil: influence of the IFNG+874 A>T polymorphism

ABSTRACT Introduction Latent tuberculosis infection diagnosis based on the release of interferon-gamma in cultures of peripheral blood cells stimulated with Mycobacterium tuberculosis antigens has replaced the tuberculin skin test in many countries with low tuberculosis prevalence. The IFN-γ production can be influenced by genetic polymorphisms, of which the IFNG + 874 (rs62559044) locus is the most studied. We investigated the possible influence of the IFNG + 874 A/T polymorphism on interferon-gamma test performance. Methods Patients diagnosed with pulmonary tuberculosis (75), volunteers with positive tuberculin skin test (70) and healthy volunteers with negative tuberculin skin test and no history of contact with tuberculosis (57) were evaluated regarding the IFNG + 874 genotype and the IFN-γ levels in whole blood cultures performed using an interferon-gamma commercial kit (QuantiFERON-TB® Gold In-Tube). Results IFN-γ production was not influenced by the IFNG + 874 genotype, regardless of antigen or mitogen-based stimulation, which suggests that other genes may influence IFN-γ production in response to mycobacteria. The IFNG + 874 polymorphism was found to exert no influence over QFT-IT test sensitivity in our study. Conclusions The IFNG + 874 polymorphism was not shown to influence QuantiFERON-TB® Gold In-Tube test performance in an admixed population from northeastern Brazil.

Rapid molecular test for tuberculosis: impact of its routine use at a referral hospital.

OBJECTIVE: To evaluate the impact of the use of the molecular test for Mycobacterium tuberculosis and its resistance to rifampin (Xpert MTB/RIF), under routine conditions, at a referral hospital in the Brazilian state of Bahia. METHODS: This was a descriptive study using the database of the Mycobacteriology Laboratory of the Octávio Mangabeira Specialized Hospital, in the city of Salvador, and georeferencing software. We evaluated 3,877 sputum samples collected from symptomatic respiratory patients, under routine conditions, between June of 2014 and March of 2015. All of the samples were submitted to sputum smear microscopy and the Xpert MTB/RIF test. Patients were stratified by gender, age, and geolocation. RESULTS: Among the 3,877 sputum samples evaluated, the Xpert MTB/RIF test detected M. tuberculosis in 678 (17.5%), of which 60 (8.8%) showed resistance to rifampin. The Xpert MTB/RIF test detected M. tuberculosis in 254 patients who tested negative for sputum smear microscopy, thus increasing the diagnostic power by 59.9%. CONCLUSIONS: The use of the Xpert MTB/RIF test, under routine conditions, significantly increased the detection of cases of tuberculosis among sputum smear-negative patients.

Interferon-gamma release assay performance in northeastern Brazil: influence of the IFNG+874 A>T polymorphism.

INTRODUCTION: Latent tuberculosis infection diagnosis based on the release of interferon-gamma in cultures of peripheral blood cells stimulated with Mycobacterium tuberculosis antigens has replaced the tuberculin skin test in many countries with low tuberculosis prevalence. The IFN-γ production can be influenced by genetic polymorphisms, of which the IFNG+874 (rs62559044) locus is the most studied. We investigated the possible influence of the IFNG+874 A/T polymorphism on interferon-gamma test performance. METHODS: Patients diagnosed with pulmonary tuberculosis (75), volunteers with positive tuberculin skin test (70) and healthy volunteers with negative tuberculin skin test and no history of contact with tuberculosis (57) were evaluated regarding the IFNG+874 genotype and the IFN-γ levels in whole blood cultures performed using an interferon-gamma commercial kit (QuantiFERON-TB® Gold In-Tube). RESULTS: IFN-γ production was not influenced by the IFNG+874 genotype, regardless of antigen or mitogen-based stimulation, which suggests that other genes may influence IFN-γ production in response to mycobacteria. The IFNG+874 polymorphism was found to exert no influence over QFT-IT test sensitivity in our study. CONCLUSIONS: The IFNG+874 polymorphism was not shown to influence QuantiFERON-TB® Gold In-Tube test performance in an admixed population from northeastern Brazil.

In vivo and in vitro expression of five genes involved in Corynebacterium pseudotuberculosis virulence.

Caseous lymphadenitis (LC) is a chronic contagious disease caused by Corynebacterium pseudotuberculosis, which mainly affects goats and sheep. Vaccination is an effective but not yet well-established method, partly due to a lack of knowledge surrounding the most effective immunoprotective components. The present study aimed to quantify and compare the in vivo expression of genes pld (phospholipase D), cpp (CP40), nanH (neuraminidase H), sodC (superoxide dismutase C) and spaC (adhesin) using qRT-PCR, with the respective expression in vitro. Caseous material of abscesses removed from five animals was cultured, with colonies suggestive of C. pseudotuberculosis identified. RNA extraction was performed on these samples, as well as on the respective pellets derived from liquid cultures brain heart infusion. After evaluating RNA integrity, complementary DNA was synthesized, followed by the relative quantification each of the genes of interest. Mean mRNA expression of the five genes found in abscesses and in cultures differed significantly, with respective values of: nanH 811.50 ± 198.27 and 359.35 ± 75.45 (p = 0.009); cpp 856.31 ± 385.11 and 154.54 ± 94.34 (p = 0.0039); plD 922.70 ± 450.73 and 212.41 ± 153.10 (p = 0.016); sodC 1,293.53 ± 564.75 and 223.63 ± 145.58 (p = 0.016); spaC 1,157.10 ± 525.13 and 214.26 ± 125.70 (p = 0,016). Expression was observed to be 6-8 times higher in abscesses than in cultures, Indicative that is a genetic expression of the in vitro bacterium exists, yet in vivo has a greater magnitude corroborating to one of these virulence factors in the pathogenesis of LC.

Specific Cytostatic and Cytotoxic Effect of Dihydrochelerythrine in Glioblastoma Cells: Role of NF-κB/ß-catenin and STAT3/IL-6 Pathways.

BACKGROUND: A glioblastoma is a primary CNS tumor that is more aggressive and lethal than other brain tumors. Its location, rapid proliferation, invasive growth, angiogenesis and immunosuppression are the main factors that limit its treatment, making it a major challenge to neuro-oncology. OBJECTIVE: This study investigated the in vitro effects of the alkaloid dihydrochelerythrine (DHC), which is extracted from Zanthoxylum stelligerum, on the viability, proliferation, cell death and ß-catenin, NFκB, STAT3/pSTAT3 and interleukins roles. METHOD: In vitro experimental models of human (U251 and GL-15) and murine (C6) glioblastoma cells were cultured in the presence of DHC at increasing concentrations for MTT assay and exclusion trypan blue dye to determine EC50. Afterward, C6 and U251 cells were treated with 100 µM DHC or DMSO 0.1% for cell cycle, annexin and expression of ß-catenin/NFκB/STAT3/pSTAT3 by flow cytometry or immunofluorescence. Interleukin quantification was made by Cytometric Bead Array. RESULTS: A significant decrease was observed in C6 and U251 cell viability in a time and dose-dependent manner. GL-15 cell viability decreased only when treated with 200 µM DHC. This maximum concentration affected neither astrocytes nor microglia viability. A cytostatic effect of DHC was observed in C6 and U251 cells after 48 h of 100 µM DHC treatment. After 72 h of DHC treatment, C6 presented 80% of annexin-V+ cells compared to 10% of annexin-V+ U251 cells. C6 cells demonstrated significant high levels of NFκ B and ß-catenin cytoplasmic fraction. Additionally, DHC treatment resulted in higher significant levels of IL-6 than did other interleukins and STAT3 up-regulation in U251 cells. CONCLUSION: These results demonstrate that DHC acts as a chemosensitizing agent selective for glioma cells not affecting non-tumor cells. Considering tumor heterogeneity, DHC demonstrated an anti-cancer potential to activate different cell death pathways. DHC demonstrated could be used for chemotherapy and immunotherapy applications in glioblastomas in the future.

Rapid molecular test for tuberculosis: impact of its routine use at a referral hospital / Teste rápido molecular para tuberculose: avaliação do impacto de seu uso na rotina em um hospital de referência

ABSTRACT Objective: To evaluate the impact of the use of the molecular test for Mycobacterium tuberculosis and its resistance to rifampin (Xpert MTB/RIF), under routine conditions, at a referral hospital in the Brazilian state of Bahia. Methods: This was a descriptive study using the database of the Mycobacteriology Laboratory of the Octávio Mangabeira Specialized Hospital, in the city of Salvador, and georeferencing software. We evaluated 3,877 sputum samples collected from symptomatic respiratory patients, under routine conditions, between June of 2014 and March of 2015. All of the samples were submitted to sputum smear microscopy and the Xpert MTB/RIF test. Patients were stratified by gender, age, and geolocation. Results: Among the 3,877 sputum samples evaluated, the Xpert MTB/RIF test detected M. tuberculosis in 678 (17.5%), of which 60 (8.8%) showed resistance to rifampin. The Xpert MTB/RIF test detected M. tuberculosis in 254 patients who tested negative for sputum smear microscopy, thus increasing the diagnostic power by 59.9%. Conclusions: The use of the Xpert MTB/RIF test, under routine conditions, significantly increased the detection of cases of tuberculosis among sputum smear-negative patients.

RESUMO Objetivo: Avaliar o impacto do teste rápido molecular automatizado Xpert MTB/RIF, utilizado para a detecção de Mycobacterium tuberculosis e sua resistência à rifampicina, em condições de rotina, em um hospital de referência no estado da Bahia. Métodos: Estudo descritivo retrospectivo utilizando o banco de dados do Laboratório de Micobacteriologia do Hospital Especializado Octávio Mangabeira, localizado na cidade de Salvador, e um programa de georreferenciamento. Entre junho de 2014 e março de 2015, foram incluídas no estudo 3.877 amostras de escarro coletadas de pacientes sintomáticos respiratórios em condições de rotina. Todas as amostras coletadas foram submetidas tanto à baciloscopia quanto a Xpert MTB/RIF. Os pacientes foram estratificados por sexo, idade e georreferenciamento. Resultados: Das 3.877 amostras de escarro analisadas, Xpert MTB/RIF detectou a presença de M. tuberculosis em 678 pacientes (17,5%). Desses, 60 (8,8%) apresentaram resistência à rifampicina. O Xpert MTB/RIF detectou 254 pacientes com baciloscopia negativa, representando um acréscimo diagnóstico de 59,9%. Conclusões: A implantação do Xpert MTB/RIF, sob condições de rotina, teve um impacto significativo no aumento da detecção de casos de tuberculose em pacientes com baciloscopia negativa.

Hepatite B na Bahia, Brasil segundo sistema de informação de agravos de notificação, 2013 / Hepatitis B in Bahia, Brazil according to the notifiable diseases surveillance system database, 2013

Objetivo: estudar os aspectos sócio-demográficos e epidemiológicos da hepatite B no estado da Bahia e avaliar a metodologia da notificação. Metodologia: em estudo descritivo foram avaliados os registros dados de pacientes do estado da Bahia notificados com hepatite B no Sistema de Informação de Agravos de Notificação. A amostra foi de 371 notificações, de janeiro a agosto de 2013. Das 52 variáveis que constituem a Ficha de Investigação (SINAN) foram selecionadas 10 variáveis de importância epidemiológica para hepatite B (HB), por relevância neste estudo, para cálculo das frequências absolutas e relativas. Resultados: em relação aos aspectos sócio-demográficos 53,4% foram do sexo masculino e a média de idade foi de 38 anos, mínima de 4 e máxima de 78 anos. As maiores fontes de transmissão foram a sexual 62,2% pessoa/pessoa 6,4% e transfusão de hemoderivados 4,8%. A variável "ignorados" correspondeu a 47,3% das notificações. Sobre a escolaridade dos infectados 22,9% eram do Ensino Médio Completo e 22,5% Fundamental Incompleto. Salvador foi o município que mais notificou casos do vírus da hepatite B (HBV) com 91, seguido por Jequié 35 e Camaçari 24. Não fizeram uso da vacina contra HBV 76,0%. Na classificação etiológica 97,3% foi de monoinfecção pelo HBV e 2,7% co-infecção com HCV. Na classificação clínica 86,9% apresentaram HB crônica e 10,4% HB aguda. Conclusão: encontrou-se pouca associação entre escolaridade e HB na população. A variável de maior fonte de transmissão registrada foi à sexual. O sexo masculino apresentou maior frequência para o HBV. Observou-se que na maioria dos notificados havia itens importantes não preenchidos.

Objective: to study the sociodemographic and epidemiological aspects of hepatitis b in the state of bahia, brazil and to evaluate the methodology involved in the notification procedure. Methodology: this descriptive study evaluated data referring to 371 patients from the state of Bahia registered as cases of hepatitis B in the national notifiable diseases surveillance system (SINAN) database between January and August 2013. Of the 52 variables included in the SINAN investigation form, 10 variables of epidemiological importance were selected and described as absolute and relative frequencies. Results: regarding the sociodemographic aspects, 53.4% of cases were male and the mean age was 38 years (range 4-78 years). Transmission was mainly sexual (62.2%),person-to ­ person (6.4%) or by transfusion of blood products (4.8%). In 47.3%, the mode of transmission was unknown. Regarding education, 22.9% of cases had completed high school, while 22.5% had failed to complete elementary school. With 91 cases, Salvador was the city with most notifications, followed by Jequié (35) and Camaçari (24).Overall, 76.0% had not been vaccinated against the hepatitis B virus (HBV). In the etiological classification, 97.3% of cases consisted of HBV mono-infection, while 2.7% were co-infected with the hepatitis C virus. Clinically, the disease was chronic in 86.9% of cases and acute in 10.4%. Conclusion: education level and hepatitis B were found to be associated in this population. The most common mode of transmission was sexual, with most cases occurring in males. In the majority of cases, important items on the notification form had not been completed.

HCV/HTLV coinfection: Does HTLV-1 interfere in the natural history of HCV-related diseases?

Hepatitis C virus (HCV) and human T-lymphotropic virus type 1 (HTLV-1) coinfection occurs in many regions. However, few studies have focused on the natural history of HCV-induced liver disease in coinfected patients. To describe the clinical, epidemiological, and histopathological aspects of HTLV-1/HCV coinfection in Brazil. A cross-sectional study with 23 patients coinfected with HCV/HTLV. The control groups consisted of 21 patients monoinfected with HCV and 20 patients monoinfected with HTLV-1. The cytokine profiles (Th1 and Th2 cell responses), clinical, laboratory features, and histopathological aspects were examined. The control group for cytokine analysis validation consisted of patients monoinfected with HTLV, and a fourth group consisted of healthy blood donors. No anthropometric differences present between the three infected groups. We observed higher serum concentrations of IFN-γ in patients coinfected with HCV/HTLV-1 than those in HCV monoinfected patients. The HCV/HTLV-1 coinfected group also exhibited a higher degree of liver steatosis than the HCV monoinfected patients. Results suggest that HCV/HTLV-1 coinfection may result in a different pattern of HCV infection due to the immunologic disorders likely associated with HTLV-1, but there is no clear evidence of the HTLV role in the natural history of HCV infection. J. Med. Virol. 88:1967-1972, 2016. © 2016 Wiley Periodicals, Inc.