RESUMO
This study aimed to investigate the toxicity of the fungicide ipconazole on oxidative status, cell death and inflammasome complex activation in the hypothalamus, cerebral cortex, striatum and hippocampus of rats. Female albino rats were randomly divided into a control group and four groups treated with ipconazole at doses of 1, 5, 10 and 20 mg/kg b.w., administered for six days. Ipconazole significantly increased MDA and ROS levels in all brain regions studied, while reducing catalase enzyme activity. The molecular expression of cell death-related genes (AKT1, APAF1, BNIP3, CASP3 and BAX) and the inflammasome complex (CASP1, IL1ß, IL6, NLRP3, NFĸB and TNFα) was also assessed, showing increased expression in at least one brain region. The findings demonstrate that ipconazole induces central nervous system toxicity in mammals, highlighting its potential role as a risk factor in the development of neurodegenerative disorders in individuals exposed to this contaminant.
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Ipconazole is an antifungal agrochemical widely used in agriculture against seed diseases of rice, vegetables, and other crops; due to its easy accumulation in the environment, it poses a hazard to human, animal, and environmental health. Therefore, we investigated the cytotoxic effect of ipconazole on SH-SY5Y neuroblastoma cells using cell viability tests (MTT), ROS production, caspase3/7 activity, and molecular assays of the biomarkers of cell death (Bax, Casp3, APAF1, BNIP3, and Bcl2); inflammasome (NLRP3, Casp1, and IL1ß); inflammation (NFκB, TNFα, and IL6); and antioxidants (NRF2, SOD, and GPx). SH-SY5Y cells were exposed to ipconazole (1, 5, 10, 20, 50, and 100 µM) for 24 h. The ipconazole, in a dose-dependent manner, reduced cell viability and produced an IC50 of 32.3 µM; it also produced an increase in ROS production and caspase3/7 enzyme activity in SH-SY5Y cells. In addition, ipconazole at 50 µM induced an overexpression of Bax, Casp3, APAF1, and BNIP3 (cell death genes); NLRP3, Casp1, and IL1B (inflammasome complex genes); and NFκB, TNFα, and IL6 (inflammation genes); it also reduced the expression of NRF2, SOD, and GPx (antioxidant genes). Our results show that ipconazole produces cytotoxic effects by reducing cell viability, generating oxidative stress, and inducing cell death in SH-SY5Y cells, so ipconazole exposure should be considered as a factor in the presentation of neurotoxicity or neurodegeneration.
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The goal of this study was to identify individuals at risk of progression and reactivation among household contacts (HHC) of pulmonary TB cases in Vitoria, Brazil. We first evaluated the predictive performance of six published signatures on the transcriptional dataset obtained from peripheral blood mononuclear cell samples from HHC that either progressed to TB disease or not (non-progressors) during a five-year follow-up. The area under the curve (AUC) values for the six signatures ranged from 0.670 to 0.461, and the PPVs did not reach the WHO published target product profiles (TPPs). We therefore used as training cohort the earliest time-point samples from the African cohort of adolescents (GSE79362) and applied an ensemble feature selection pipeline to derive a novel 29-gene signature (PREDICT29). PREDICT29 was tested on 16 progressors and 21 non-progressors. PREDICT29 performed better in segregating progressors from non-progressors in the Brazil cohort with the area under the curve (AUC) value of 0.911 and PPV of 20%. This proof of concept study demonstrates that PREDICT29 can predict risk of progression/reactivation to clinical TB disease in recently exposed individuals at least 5 years prior to disease development. Upon validation in larger and geographically diverse cohorts, PREDICT29 can be used to risk-stratify recently infected for targeted therapy.
Assuntos
Perfilação da Expressão Gênica , Tuberculose Latente/diagnóstico , Mycobacterium tuberculosis/patogenicidade , Transcriptoma , Tuberculose Pulmonar/diagnóstico , África , Brasil , Estudos de Casos e Controles , Busca de Comunicante , Progressão da Doença , Características da Família , Interações Hospedeiro-Patógeno , Humanos , Tuberculose Latente/genética , Tuberculose Latente/microbiologia , Tuberculose Latente/transmissão , Valor Preditivo dos Testes , Estudo de Prova de Conceito , Estudos Prospectivos , Reinfecção , Reprodutibilidade dos Testes , Medição de Risco , Fatores de Risco , Fatores de Tempo , Tuberculose Pulmonar/genética , Tuberculose Pulmonar/microbiologia , Tuberculose Pulmonar/transmissãoRESUMO
BACKGROUND: Mycobacterium tuberculosis cultures of cough-generated aerosols from patients with pulmonary tuberculosis (TB) are a quantitative method to measure infectiousness and to predict secondary outcomes in exposed contacts. However, their reproducibility has not been established. OBJECTIVE: To evaluate the predictive value of colony-forming units (CFU) of M. tuberculosis in cough aerosols on secondary infection and disease in household contacts in Brazil. METHODS: Adult sputum smear+ and culture+ pulmonary TB cases underwent a standard evaluation and were categorized according to aerosol CFU. We evaluated household contacts for infection at baseline and at 8 weeks with TST and IGRA, and secondary disease. RESULTS: We enrolled 48 index TB cases; 40% had negative aerosols, 27% low aerosols (<10 CFU) and 33% high aerosols (≥10 CFU). Of their 230 contacts, the proportion with a TST ≥10 mm at 8 weeks was 59%, 65% and 75%, respectively (p = 0.34). Contacts of high aerosol cases had greater IGRA readouts (median 4.6 IU/mL, IQR 0.02-10) when compared to those with low (0.8, 0.2-10) or no aerosol (0.1, 0-3.7; p = 0.08). IGRA readouts in TST converters of high aerosol cases (median 20 IU/mL, IQR 10-24) were larger than those from aerosol-negative (0.13, 0.04-3; p = o.o2). 8/9 (89%) culture+ secondary TB cases occurred in contacts of aerosol+ cases. CONCLUSION: Aerosol CFU predicts quantitatively IGRA readouts among household contacts of smear positive TB cases. Our results strengthen the argument of using cough aerosols to guide targeted preventive treatment strategies, a necessary component of current TB elimination projections.
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Tosse/microbiologia , Habitação , Mycobacterium tuberculosis/fisiologia , Tuberculose Pulmonar/microbiologia , Tuberculose Pulmonar/transmissão , Adulto , Aerossóis , Brasil , Técnicas de Cultura , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mycobacterium tuberculosis/crescimento & desenvolvimento , Valor Preditivo dos TestesRESUMO
Household contacts of pulmonary tuberculosis (TB) patients are at increased risk of TB infection and disease. However, their risk in relation to the intensity of exposure remains unknown.We studied smear-positive TB cases and their household contacts in Vitória, Brazil. We collected clinical, demographic and radiographic information from TB cases, and obtained tuberculin skin test (TST) and QuantiFERON-TB Gold (QFT) results from household contacts. We measured intensity of exposure using a proximity score and sleep location in relation to the TB index case and defined infection by TST ≥10â mm or QFT ≥0.35â UI·mL-1 We ascertained secondary TB cases by reviewing local and nationwide case registries.We included 160 TB index cases and 894 household contacts. 464 (65%) had TB infection and 23 (2.6%) developed TB disease. Risk of TB infection and disease increased with more intense exposures. In an adjusted analysis, the proximity score was associated with TB disease (OR 1.61, 95% CI 1.25-2.08; p<0.000); however, its diagnostic performance was only moderate.Intensity of exposure increased risk of TB infection and disease among household contacts; however, its diagnostic performance was still suboptimal. A biomarker to target preventive therapy is urgently needed in this at-risk population.
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Busca de Comunicante/métodos , Tuberculose Pulmonar/transmissão , Adulto , Área Sob a Curva , Biomarcadores/metabolismo , Brasil , Controle de Doenças Transmissíveis , Características da Família , Feminino , Humanos , Infectologia/métodos , Testes de Liberação de Interferon-gama , Masculino , Pessoa de Meia-Idade , Mycobacterium tuberculosis , Curva ROC , Risco , Teste Tuberculínico/métodos , Tuberculose Pulmonar/epidemiologiaRESUMO
BACKGROUND: In household contact investigations of tuberculosis (TB), a second tuberculin skin test (TST) obtained several weeks after a first negative result consistently identifies individuals that undergo TST conversion. It remains unclear whether this delay in M. tuberculosis infection is related to differences in the infectious exposure, TST boosting, partial host resistance, or some other factor. METHODS: We conducted a household contact study Vitória, Brazil. Between 2008 and 2013, we identified culture-positive pulmonary TB patients and evaluated their household contacts with both a TST and interferon gamma release assay (IGRA), and identified TST converters at 8-12 weeks post study enrollment. Contacts were classified as TST-positive (≥10 mm) at baseline, TST converters, or persistently TST-negative. We compared TST converters to TST-positive and to TST-negative contacts separately, using generalized estimating equations. RESULTS: We enrolled 160 index patients and 838 contacts; 523 (62.4%) were TST+, 62 (7.4%) TST converters, and 253 (30.2%) TST-. TST converters were frequently IGRA- at 8-12 weeks. In adjusted analyses, characteristics distinguishing TST converters from TST+ contacts (no contact with another TB patient and residence ownership) were different than those differentiating them from TST- contacts (stronger cough in index patient and contact BCG scar). CONCLUSIONS: The individual risk and timing of M. tuberculosis infection within households is variable and dependent on index patient, contact and environmental factors within the household, and the surrounding community. Our findings suggest a threshold effect in the risk of infection in humans.
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Tuberculose/diagnóstico , Tuberculose/epidemiologia , Adolescente , Adulto , Brasil/epidemiologia , Criança , Tosse/microbiologia , Características da Família , Feminino , Humanos , Testes de Liberação de Interferon-gama , Masculino , Mycobacterium tuberculosis/patogenicidade , Teste Tuberculínico , Tuberculose/transmissão , Tuberculose Pulmonar/diagnóstico , Tuberculose Pulmonar/epidemiologia , Adulto JovemRESUMO
In the United States, multidrug-resistant tuberculosis (MDR-TB) is more commonly seen among foreign-born patients. We report outcomes for 46 patients with MDR-TB who were born in Mexico and treated along the United States-Mexico border. According to our definition, 30 were cured, 3 showed treatment failure, 3 died, and 10 abandoned treatment. Multidrug-resistant tuberculosis can be successfully treated on an ambulatory basis.
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Antituberculosos/uso terapêutico , Americanos Mexicanos , Tuberculose Resistente a Múltiplos Medicamentos/tratamento farmacológico , Tuberculose Resistente a Múltiplos Medicamentos/epidemiologia , Tuberculose Pulmonar/tratamento farmacológico , Tuberculose Pulmonar/epidemiologia , Assistência Ambulatorial , Estudos de Coortes , Esquema de Medicação , Farmacorresistência Bacteriana Múltipla , Feminino , Humanos , Masculino , México/etnologia , Pessoa de Meia-Idade , Texas/epidemiologia , Falha de Tratamento , Tuberculose Resistente a Múltiplos Medicamentos/etnologia , Tuberculose Pulmonar/etnologiaRESUMO
OBJECTIVE: To evaluate the diagnostic performance of two liquid-phase culture media for the diagnosis of pulmonary tuberculosis. PATIENTS AND METHODS: From May to July 2003, sputum samples for culture were obtained from patients with respiratory symptoms attending the Hospital Nacional Cayetano Heredia. These were cultured in Ogawa medium, mycobacteria growth indicator tube (MGIT), and modified Middlebrook 7H9. Results were compared against a composite reference standard. RESULTS: One hundred sputum specimens from 100 patients were included. Of these, 33 had culture-proven tuberculosis. The sensitivity of MGIT was found to be 100%. The modified Middlebrook 7H9 medium was found to have a sensitivity of 72.73%, while the sensitivity of Ogawa medium was found to be 69.70%. The mean growing time for MGIT was 12.18 days (95% confidence interval 10.24 to 14.12; p<0.01 vs. Ogawa and modified Middlebrook 7H9); for modified Middlebrook 7H9 was 16.65 days (95% confidence interval 14.85 to 18.80; p<0.01 vs. Ogawa), and for the Ogawa medium 25.74 days (95% confidence interval 22.22 to 29.6). CONCLUSIONS: The liquid culture medium MGIT was superior to the modified Middlebrook 7H9 and the Ogawa media, both in terms of sensitivity and shorter growing time of colonies of Mycobacterium tuberculosis. The modified Middlebrook 7H9 medium is significantly faster but comparable in diagnostic performance to Ogawa. Costs remain an issue for MGIT.
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Meios de Cultura , Mycobacterium tuberculosis/isolamento & purificação , Tuberculose Pulmonar/diagnóstico , Adolescente , Adulto , Técnicas Bacteriológicas , Hospitais Públicos , Humanos , Peru , Sensibilidade e Especificidade , Escarro/microbiologia , Tuberculose Pulmonar/microbiologia , Adulto JovemRESUMO
This study developed a clinical score based on clinical and radiographic data for the diagnosis of smear-negative pulmonary tuberculosis (SNPT). SNPT was defined as a positive culture in Ogawa in a patient with two negative sputum smears. Data from patients admitted to the emergency ward with respiratory symptoms and negative acid-fast bacilli (AFB) smears was analyzed by means of logistic regression to develop the predictive score.Two hundred and sixty two patients were included. Twenty patients had SNPT. The variables included in the final model were hemoptysis, weight loss, age > 45 years old, productive cough, upper-lobe infiltrate, and miliary infiltrate. With those, a score was constructed. The score values ranged from -2 to 6. The area under the curve for the ROC curve was 0.83 (95% CI 0.74-0.90). A score of value 0 or less was associated with a sensitivity of 93% and a score of more than 4 points was associated with a specificity of 92% for SNPT. Fifty-two point twenty-nine percent of patients had scores of less than one or more than four, what provided strong evidence against and in favor, respectively, for the diagnosis of SNPT. The score developed is a cheap and useful clinical tool for the diagnosis of SNPT and can be used to help therapeutic decisions in patients with suspicion of having SNPT.
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Tuberculose Pulmonar/diagnóstico , Fatores Etários , Tosse/etiologia , Métodos Epidemiológicos , Feminino , Hemoptise/etiologia , Humanos , Masculino , Pessoa de Meia-Idade , Peru , Radiografia , Tuberculose Pulmonar/diagnóstico por imagem , Redução de PesoRESUMO
BACKGROUND: Resistance to commonly used antituberculosis drugs is emerging worldwide. Conventional drug-susceptibility testing (DST) methods are slow and demanding. Alternative, rapid DST methods would permit the early detection of drug resistance and, in turn, arrest tuberculosis transmission. METHODS: A cost-effectiveness analysis of 5 DST methods was performed in the context of a clinical trial that compared rapid with conventional DST methods. The methods under investigation were direct phage-replication assay (FASTPlaque-Response; Biotech), direct amplification and reverse hybridization of the rpoB gene (INNO-LiPA; Innogenetics), indirect colorimetric minimum inhibitory concentration assay (MTT; ICN Biomedicals), and direct proportion method on Löwenstein-Jensen medium. These were compared with the widely used indirect proportion method on Löwenstein-Jensen medium. RESULTS: All alternative DST methods were found to be cost-effective, compared with other health care interventions. DST methods also generate substantial cost savings in settings of high prevalence of multidrug-resistant tuberculosis. Excluding the effects of transmission, the direct proportion method on Löwenstein-Jensen medium was the most cost-effective alternative DST method for patient groups with prevalences of multidrug-resistant tuberculosis of 2%, 5%, 20%, and 50% (cost in US$2004, $94, $36, $8, and $2 per disability-adjusted life year, respectively). CONCLUSION: Alternative, rapid methods for DST are cost-effective and should be considered for use by national tuberculosis programs in middle-income countries.
Assuntos
Antituberculosos/farmacologia , Farmacorresistência Bacteriana Múltipla , Mycobacterium tuberculosis/efeitos dos fármacos , Tuberculose Resistente a Múltiplos Medicamentos/diagnóstico , Proteínas de Bactérias/genética , Colorimetria , Análise Custo-Benefício , RNA Polimerases Dirigidas por DNA , Amplificação de Genes , Humanos , Renda/classificação , Testes de Sensibilidade Microbiana/economia , Testes de Sensibilidade Microbiana/métodos , Micobacteriófagos/fisiologia , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/crescimento & desenvolvimento , Hibridização de Ácido Nucleico , Peru , Anos de Vida Ajustados por Qualidade de Vida , Kit de Reagentes para Diagnóstico , Sensibilidade e Especificidade , Fatores de Tempo , Tuberculose Resistente a Múltiplos Medicamentos/epidemiologia , Tuberculose Resistente a Múltiplos Medicamentos/microbiologia , Tuberculose Pulmonar/diagnóstico , Tuberculose Pulmonar/epidemiologia , Tuberculose Pulmonar/microbiologiaRESUMO
This study developed a clinical score based on clinical and radiographic data for the diagnosis of smear-negative pulmonary tuberculosis (SNPT). SNPT was defined as a positive culture in Ogawa in a patient with two negative sputum smears. Data from patients admitted to the emergency ward with respiratory symptoms and negative acid-fast bacilli (AFB) smears was analyzed by means of logistic regression to develop the predictive score.Two hundred and sixty two patients were included. Twenty patients had SNPT. The variables included in the final model were hemoptysis, weight loss, age > 45 years old, productive cough, upper-lobe infiltrate, and miliary infiltrate. With those, a score was constructed. The score values ranged from -2 to 6. The area under the curve for the ROC curve was 0.83 (95 percent CI 0.74-0.90). A score of value 0 or less was associated with a sensitivity of 93 percent and a score of more than 4 points was associated with a specificity of 92 percent for SNPT. Fifty-two point twenty-nine percent of patients had scores of less than one or more than four, what provided strong evidence against and in favor, respectively, for the diagnosis of SNPT. The score developed is a cheap and useful clinical tool for the diagnosis of SNPT and can be used to help therapeutic decisions in patients with suspicion of having SNPT.