BACKGROUND The Bacille Calmette-Guérin (BCG) vaccine comprises a family of strains with variable protective efficacy against pulmonary tuberculosis (TB) and leprosy, partly due to genetic differences between strains. OBJECTIVES Previous data highlighting differences between the genomes and proteomic profiles of BCG strains Moreau and Pasteur led us to evaluate their behaviour in the macrophage microenvironment, capable of stimulating molecular responses that can impact the protective effect of the vaccine. METHODS Strain infectivity, viability, co-localisation with acidified vesicles, macrophage secretion of IL-1 and MCP-1 and lipid droplet biogenesis were evaluated after infection. FINDINGS We found that BCG Moreau is internalised more efficiently, with significantly better intracellular survival up to 96 h p.i., whereas more BCG Pasteur bacilli were found co-localised in acidified vesicles up to 6 h p.i. IL-1β and MCP-1 secretion and lipid droplet biogenesis by infected macrophages were more prominent in response to BCG Pasteur. MAIN CONCLUSION Overall, our results show that, compared to Pasteur, BCG Moreau has increased fitness and better endurance in the harsh intracellular environment, also regulating anti-microbial responses (lower IL-1b and MCP-1). These findings contribute to the understanding of the physiology of BCG Moreau and Pasteur in response to the intraphagosomal environment in a THP-1 macrophage model.

BACKGROUND Leprosy, caused by Mycobacterium leprae, is a public health problem in Brazil that affects peripheral nerves, resulting in physical disabilities. During host-pathogen interactions, the immune response determines leprosy outcomes from a localised (paucibacillary) form to a disseminated (multibacillary) form. The recognition of M. leprae involves the DC-SIGN receptor, which is present on the dendritic cells (DCs) and participates in immune activation. OBJECTIVES To evaluate the association of polymorphisms in the promoter region of the gene encoding DC-SIGN (CD209) and the clinical form of leprosy, and to investigate its functional effects. METHODS The study population included 406 leprosy patients from an endemic area in Brazil [310 multibacillary (MB); 96 paucibacillary (PB)]. A functional evaluation based on the effects of the single nucleotide variant (SNV) associated with PB leprosy on the specific immune response was also performed. RESULTS The GA genotype and the presence of the A allele of rs735240 (-939G>A) were associated with PB leprosy [OR: 2.09 (1.18-3.69) and 1.84 (1.07-3.14), respectively]. Carriers of the A allele showed reduced expression of CD209 and TGF-β1 in leprosy lesions in comparison with individuals with GG genotype, in addition to a higher response to the Mitsuda test. CONCLUSION These data suggest that rs735240 influences the immune response against M. leprae and clinical presentation of leprosy.

BACKGROUND Leprosy is curable by multidrug therapy (MDT) treatment regimen ranging from six to 12 months. The variable levels of tolerance and adherence among patients can, however, result in treatment failure and the emergence of drug-resistant strains. OBJECTIVES Describe the impact of MDT over Mycobacterium leprae viability in patient’s oral and nasal mucosa along treatment. METHODS Mycobacterium leprae viability was monitored by quantitative polymerase chain reaction (qPCR) quantification of 16S rRNA in lateral and contralateral scrapings of oral and nasal mucosa of 10 multibacillary patients along the initial five months of treatment. FINDINGS The results demonstrated high heterogenicity of M. leprae viability among patients and between nasal and oral samples. Of six patients who presented good adherence and tolerance to the treatment, only four displayed absence of M. leprae viability in both samples three months after the first MDT dose, while for the other two, the absence of M. leprae viability in the oral and nasal cavities was only detected five months after the first dose. MAIN CONCLUSIONS We concluded that qPCR of 16S rRNA for the determination of M. leprae viability in nasal and oral scraping samples could represent an interesting approach to monitor treatment efficacy.

Abstract INTRODUCTION: Leprosy recurrence is the reappearance of the disease after treatment with current schemes and discharged for cure and may have variable incubation periods. METHODS: This is a descriptive observational study of leprosy recurrence in Espírito Santo diagnosed between January 2018 and January 2020. RESULTS: One hundred and ninety-two cases were available, of which 30 were diagnosed with leprosy recurrence. CONCLUSIONS: In 25 cases, the incubation period was 5-15 years after the first treatment, favoring bacillary persistence. In the remaining 5 cases, the disease had recurred after 15 years, pointing to reinfection as none of them exhibited drug resistance.

BACKGROUND Occult hepatitis B virus (HBV) - characterized by the absence of detectable HBsAg in the presence of HBV DNA - represents a potential threat for blood safety. OBJECTIVES This study was conducted with the aim to investigate the serological and molecular characterization of occult HBV infection (OBI) among blood donors in Mozambique. METHODS 1,502 blood donors were tested for HBsAg. All HBsAg-negative individuals were tested for HBV DNA. Antibodies against HBV core, surface and HBe antigen (anti-HBc, anti-HBs, HBeAg) were measured in HBV DNA positive individuals. FINDINGS 1435 serum samples were HBsAg negative and 16 positive for HBV DNA, 14 confirmed to have OBI, corresponding to a frequency of 0.98%. Of the 14 OBI infections identified, 13/14 (92.8%) were positive for anti-HBc, 4/14 (28.5%) for anti-HBs, and no samples were reactive for HBeAg. Of the 14 OBI cases, nine samples (64.2%) were sequenced for the S/P region. Eight samples (88.9%) belonged to genotype A1 and one (11.1%) to genotype E. One escape mutation (T123A) associated with OBI and various amino acid substitutions for genotype A1 and E were observed. MAIN CONCLUSIONS Our results show the importance of using nucleic acid amplification test to detect occult hepatitis B infection in blood donors in Mozambique.

RESUMO O estado de Mato Grosso, com uma área de 903 357 km², não possui uma metodologia oficial para estimar a acidez potencial do solo (H + Al), e sua determinação pelo método padrão é onerosa e demorada. O objetivo deste estudo foi comparar os valores estimados de (H + Al) determinados pelo método padrão de acetato de cálcio com os obtidos por três métodos tampão, a saber, tampão Shoemaker, McLean and Pratt (SMP), tampão Sikora e tampão Santa Maria (SMB), considerando amostras das principais classes de solos cultivados no estado de Mato Grosso. Para isso, 196 amostras de solo foram coletadas na camada arável (0-20 cm) em diferentes sistemas agropastoris e nativos adjacentes. Os modelos estatísticos obtidos foram comparados com modelos hipoteticamente inadequados utilizados pelos laboratórios, pois não há calibração para solos no estado. Após as análises laboratoriais, os pares de valores determinados de H+Al e o pH de equilíbrio corresponde de cada solução tampão foram submetidos a análise de correlação e regressão não-linear (p < 0,05). Verificou-se que o método SMB, que não gera resíduos de poluentes no meio ambiente, foi melhor que os métodos Sikora e SMP para substituir o método padrão, usando a equação H + Al (cm 3 c dm-3) = 51.189-25.70 ln (pHSMB) (R2 = 0,88, P < 0,0001). Assim, se os laboratórios usarem equações não calibradas para estimar a acidez potencial do solo, isso implica subestimar ou superestimar a recomendação de calcário, o que pode comprometer a produtividade das culturas no Mato Grosso.

ABSTRACT Mato Grosso, with a total area of 903357 km², does not have an official methodology for estimating soil potential acidity (H + Al), and determination of H + Al using the standard method is onerous and time consuming. The objective of this study was to compare estimated values of H + Al determined using the standard calcium acetate method with those obtained using three buffer methods, namely, the Shoemaker, McLean, and Pratt (SMP) buffer, Sikora buffer, and Santa Maria buffer (SMB) methods, with samples of the main classes of cultivated soils in the state of Mato Grosso. A total of 196 soil samples were collected from the arable layer (0-20 cm) in agropastoral and adjacent native systems. Statistical models were obtained and compared with models used by laboratories that are hypothetically inadequate because there is no calibration for soils in the state. After laboratory analyses, the paired H + Al and equilibrium pH values corresponding to the equilibrium of each buffer solution (SMP buffer, SMB, and Sikora buffer) were subjected to nonlinear regression analysis (P < 0.05). The SMB method, which does not release pollutant residues into the environment, was better than the Sikora and SMP methods for replacing the standard method used in state laboratories for soil analysis, that is, H + Al (cm3 c dm-3) = 51.189 -25.70 ln(pHSMB) (R2 = 0.88, P < 0.0001). Thus, if laboratories use uncalibrated equations to estimate soil potential acidity, the recommended limestone correction will be underestimated or overestimated, which may compromise crop productivity in Mato Grosso.

Abstract INTRODUCTION: Chagas disease (CD) is a neglected disease caused by the parasite Trypanosoma cruzi. One-third of infected patients will develop the cardiac form, which may progress to heart failure (HF). However, the factors that determine disease progression remain unclear. Increased angiotensin II activity is a key player in the pathophysiology of HF. A functional polymorphism of the angiotensin-converting enzyme (ACE) gene is associated with plasma enzyme activity. In CD, ACE inhibitors have beneficial effects supporting the use of this treatment in chagasic cardiomyopathy. METHODS: We evaluated the association of ACE I/D polymorphism with HF, performing a case-control study encompassing 343 patients with positive serology for CD staged as non-cardiomyopathy (stage A; 100), mild (stage B1; 144), and severe (stage C; 99) forms of Chagas heart disease. For ACE I/D genotyping by PCR, groups were compared using unconditional logistic regression analysis and adjusted for nongenetic covariates: age, sex, and trypanocidal treatment. RESULTS: A marginal, but not significant (p=0.06) higher prevalence of ACE I/D polymorphism was observed in patients in stage C compared with patients in stage A. Patients in stage C (CD with HF), were compared with patients in stages A and B1 combined into one group (CD without HF); DD genotype/D carriers were prevalent in the HF patients (OR = 2; CI = 1.013.96; p = 0.04). CONCLUSIONS: Our results of this cohort study, comprising a population from the Northeast region of Brazil, suggest that ACE I/D polymorphism is more prevalent in the cardiac form of Chagas disease with HF.

BACKGROUND Dengue is an arthropod-borne viral disease with a majority of asymptomatic individuals and clinical manifestations varying from mild fever to severe and potentially lethal forms. An increasing number of genetic studies have outlined the association between host genetic variations and dengue severity. Genes associated to viral recognition and entry, as well as those encoding mediators of the immune response against infection are strong candidates for association studies. OBJECTIVES The aim of this study was to investigate the association between MBL2, CLEC5A, ITGB3 and CCR5 genes and dengue severity in children. METHODS A matched case-control study was conducted and 19 single nucleotide polymorphisms (SNPs) were investigated. FINDINGS No associations were observed in single SNP analysis. However, when MBL2 SNPs were combined in haplotypes, the allele rs7095891G/rs1800450C/ rs1800451C/rs4935047A/rs930509G/rs2120131G/rs2099902C was significantly associated to risk of severe dengue under α = 0.05 (aOR = 4.02; p = 0.02). A second haplotype carrying rs4935047G and rs7095891G alleles was also associated to risk (aOR = 1.91; p = 0.04). MAIN CONCLUSIONS This is the first study to demonstrate the association between MBL2 haplotypes and dengue severity in Brazilians including adjustment for genetic ancestry. These results reinforce the role of mannose binding lectin in immune response to DENV.

BACKGROUND The severity of chronic chagasic cardiomyopathy (CCC), the most frequent clinical outcome of Chagas disease (CD), has been associated with cytokine-enriched heart tissue inflammation, and high serum levels of transforming growth factor (TGFβ), interferon-gamma (IFNγ), and tumour necrosis factor (TNF). Conversely, increased interleukin (IL)-10 serum concentrations have been associated with asymptomatic CD. Cytokines and cytokine-related gene polymorphisms may control cytokine expression and have been proposed to contribute to CCC outcomes. OBJECTIVES We evaluated the association of 13 cytokine-related genes (TGFB: rs8179181, rs8105161, rs1800469; IL10: rs1800890, rs1800871, rs1800896; IFNG: rs2430561; TNF: rs1800629; BAT1: rs3853601; LTA: rs909253, rs2239704; TNFR1: rs767455; TNFR2: rs1061624) with risk and progression of CCC. FINDINGS Four hundred and six seropositive patients from CD endemic areas in the state of Pernambuco, north-eastern Brazil, were classified as non-cardiopathic (A, 110) or cardiopathic (mild, B1, 163; severe, C, 133). We found no evidence of TGFB, IL10, TNF, or TNFR1/2 gene polymorphisms associated with CCC risk or progression. Only BAT1 rs3853601 −22G carriers (B1 vs. C: OR = 0.5; p-value = 0.03) and IFNG rs2430561 +874AT (A vs. C: OR = 0.7; p-value = 0.03; A vs. B1+C: OR = 0.8; p-value = 0.02) showed a significant association with protection from cardiopathy in a logistic regression analysis with adjustment for gender and ethnicity; however, the association disappeared after performing adjustment for multiple testing. A systematic review of TNF rs1800629 −308G>A publications included five studies for meta-analysis (534 CCC and 472 asymptomatic patients) and showed no consensus in pooled odds ratio (OR) estimates for A allele or A carriers (OR = 1.4 and 1.5; p-values = 0.14 and 0.15, respectively). In CD patients, TNF serum levels were increased, but not affected by the TNF rs1800629 −308A allele. MAIN CONCLUSIONS Our data suggest no significant contribution of the analysed gene variants of cytokine-related molecules to development/severity of Chagas' heart disease, reinforcing the idea that parasite/host interplay is critical to CD outcomes.

RESUMO Avaliou-se, neste trabalho, o efeito de um fertilizante organomineral enriquecido com substâncias húmicas em soja submetida a estresse hídrico. Em casa de vegetação foi realizado um experimento utilizando-se, como substrato, um Latossolo Vermelho com nível de fertilidade adequado no qual foram cultivadas plantas de soja em duas condições, sem e com estresse hídrico. O delineamento foi em blocos casualizados em esquema fatorial 2 x 5 (dois níveis de umidade e cinco doses: 0, 1, 2, 4 e 8 mL dm-3) totalizando 10 tratamentos, com 4 repetições. O fertilizante organomineral foi aplicado ao solo 21 dias após a emergência das plantas estabelecendo-se, uma semana após sua aplicação, os níveis de umidade. O fertilizante organomineral não foi capaz de atenuar os efeitos adversos do estresse hídrico reduzindo a produtividade dos grãos de soja em mais de 50% quando comparado às plantas cultivadas sem estresse hídrico. Houve resposta positiva quanto às doses sobre a nutrição e produtividade de grãos sendo que, na condição de estresse hídrico, a mais eficiente foi 5,4 mL dm-3; foi menor a concentração foliar de nitrogênio, fósforo e potássio nas plantas com estresse e maior concentração de enxofre; as substâncias húmicas favorecem a absorção de micronutrientes.

ABSTRACT This study evaluated the effect of an organomineral fertilizer enriched with humic substances on soybean grown under water stress. The experiment was performed in a greenhouse using a Red Latosol (Oxisol) with adequate fertility as substrate, in which soybean plants were cultivated with and without water stress. The experimental design was randomized blocks, in a 2 x 5 factorial scheme (two moisture levels and five fertilizer doses: 0, 1, 2, 4 and 8 mL dm-3), totaling 10 treatments, with four replicates. The organomineral fertilizer was applied in the soil 21 days after plant emergence and the water regimes were established one week thereafter. The fertilizer was not able to attenuate the effects of water stress, reducing soybean grain yield by more than 50% compared with plants cultivated under no stress. Fertilizer doses caused positive response on soybean nutrition and grain yield and, under water stress condition, the most efficient dose was 5.4 mL dm-3. There were lower leaf concentrations of nitrogen, phosphorus and potassium and higher concentrations of sulfur in plants under stress. Humic substances favor the absorption of micronutrients.

Resumo Fundamentos: O treinamento de força (TF) tem sido recomendado como tratamento não farmacológico para hipertensão arterial moderada. Apesar do papel importante que a intensidade do exercício desempenha sobre a prescrição do treinamento, ainda não há nenhum dado avaliando os efeitos da intensidade do TF sobre a hipertensão arterial grave (HAG). Objetivo: Este estudo analisou os efeitos de dois protocolos do TF(subida em escada vertical), realizados com diferentes sobrecargas do peso máximo carregado (PMC), sobre a pressão arterial (PA) e a força muscular de ratos espontaneamente hipertensos (SHR) com HAG. Métodos: Quinze SHR machos (206 ± 10 mmHg de PA sistólica (PAS)) e cinco ratos Wistar Kyoto (WKY; 119 ± 10 mmHg de PAS) foram divididos em 4grupos:sedentários: (SED-WKY) e SHR (SED-SHR); treinados: TF1-SHR conforme o peso corporal (~40% do PMC); e TF2-SHR conforme o teste de PMC (~70% do PMC). Foram coletadas medidas de PAS e a frequência cardíaca (FC) semanalmente usando o método de pressão arterial caudal. A progressão da força muscular foi determinada a cada 15 dias. O TF consistiu de 3 sessões semanais em dias não consecutivos durante 12 semanas. Resultados: Os dois protocolos de TF preveniram o aumento da PAS(respectivamente, delta - 5 e -7 mmHg; p > 0, 05), enquanto que a PAS do grupo SED-SHR aumentou em 19 mmHg (p < 0, 05). Houve queda na FC apenas para o grupo TF1 (p < 0, 05). Foi observado um aumento mas significativo de força no grupo do protocolo TF2 (140%; p < 0, 05) em comparação com o TF1 (11%; p>0, 05). Conclusões: Nossos dados indicam que ambos os protocolos de TF foram efetivos na prevenção da elevação crônica da PAS na HAG. Além disso, sobrecargas maiores de TF induziram a um maior aumento de força muscular.

Abstract Background: Resistance training (RT) has been recommended as a non-pharmacological treatment for moderate hypertension. In spite of the important role of exercise intensity on training prescription, there is still no data regarding the effects of RT intensity on severe hypertension (SH). Objective: This study examined the effects of two RT protocols (vertical ladder climbing), performed at different overloads of maximal weight carried (MWC), on blood pressure (BP) and muscle strength of spontaneously hypertensive rats (SHR) with SH. Methods: Fifteen male SHR ENT#091;206 ± 10 mmHg of systolic BP (SBP)ENT#093; and five Wistar Kyoto rats (WKY; 119 ± 10 mmHg of SBP) were divided into 4 groups: sedentary (SED-WKY) and SHR (SED-SHR); RT1-SHR training relative to body weight (~40% of MWC); and RT2-SHR training relative to MWC test (~70% of MWC). Systolic BP and heart rate (HR) were measured weekly using the tail-cuff method. The progression of muscle strength was determined once every fifteen days. The RT consisted of 3 weekly sessions on non-consecutive days for 12-weeks. Results: Both RT protocols prevented the increase in SBP (delta - 5 and -7 mmHg, respectively; p > 0.05), whereas SBP of the SED-SHR group increased by 19 mmHg (p < 0.05). There was a decrease in HR only for the RT1 group (p < 0.05). There was a higher increase in strength in the RT2 (140%; p < 0.05) group as compared with RT1 (11%; p > 0.05). Conclusions: Our data indicated that both RT protocols were effective in preventing chronic elevation of SBP in SH. Additionally, a higher RT overload induced a greater increase in muscle strength.

Em comparação a outros micronutrientes, os teores de níquel (Ni) disponíveis no solo e tecido vegetal são bastante baixos, com alto grau de dificuldade para a quantificação deles. O objetivo deste estudo foi apresentar métodos otimizados de determinação da disponibilidade de Ni em solos com extratores e teor total em tecido vegetal para análises de rotina em laboratórios comerciais. Foram processadas e analisadas amostras de solos naturais e agrícolas com extração pelo extrator Mehlich-1 e por DTPA. Para quantificar o Ni no tecido vegetal, as amostras foram digeridas com ácido nítrico em sistema fechado em forno micro-ondas. A determinação foi realizada em espectrômetro de emissão óptica com plasma induzido (ICP-OES). Houve correlação positiva e significativa entre os teores disponíveis de Ni nos solos extraídos em Mehlich-1 e DTPA, enquanto, para o tecido vegetal os teores de Ni recuperados foram altos e similares aos materiais de referência. A disponibilidade de Ni em algumas amostras de solos naturais e tecido vegetal foram inferiores aos limites de quantificação. Os teores desse micronutriente foram maiores em amostras de solo em que houve fornecimento de Ni. A concentração de Ni difere nas partes vegetais analisadas, e os grãos de soja apresentaram os maiores teores. Os grãos, em relação às partes aérea e foliar, têm melhor correlação com os teores disponíveis no solo para ambos extratores. Os métodos descritos nesta pesquisa foram eficientes na quantificação de Ni e podem ser adotados em laboratórios de rotina de solos e tecido vegetal.

In comparison with other micronutrients, the levels of nickel (Ni) available in soils and plant tissues are very low, making quantification very difficult. The objective of this paper is to present optimized determination methods of Ni availability in soils by extractants and total content in plant tissues for routine commercial laboratory analyses. Samples of natural and agricultural soils were processed and analyzed by Mehlich-1 extraction and by DTPA. To quantify Ni in the plant tissues, samples were digested with nitric acid in a closed system in a microwave oven. The measurement was performed by inductively coupled plasma/optical emission spectrometry (ICP-OES). There was a positive and significant correlation between the levels of available Ni in the soils subjected to Mehlich-1 and DTPA extraction, while for plant tissue samples the Ni levels recovered were high and similar to the reference materials. The availability of Ni in some of the natural soil and plant tissue samples were lower than the limits of quantification. Concentrations of this micronutrient were higher in the soil samples in which Ni had been applied. Nickel concentration differed in the plant parts analyzed, with highest levels in the grains of soybean. The grain, in comparison with the shoot and leaf concentrations, were better correlated with the soil available levels for both extractants. The methods described in this article were efficient in quantifying Ni and can be used for routine laboratory analysis of soils and plant tissues.

The diagnosis of single-lesion paucibacillary leprosy remains a challenge. Reviews by expert dermatopathologists and quantitative polymerase chain reaction (qPCR) results obtained from 66 single-plaque biopsy samples were compared. Histological findings were graded as high (HP), medium (MP) or low (LP) probability of leprosy or other dermatopathy (OD). Mycobacterium leprae-specific genes were detected using qPCR. The biopsies of 47 out of 57 clinically diagnosed patients who received multidrug therapy were classified as HP/MP, eight of which were qPCR negative. In the LP/OD (n = 19), two out of eight untreated patients showed positive qPCR results. In the absence of typical histopathological features, qPCR may be utilised to aid in final patient diagnosis, thus reducing overtreatment and delay in diagnosis.