The rheumatic heart disease healthcare paradox: disease persistence in slums despite universal healthcare coverage-a provider perspective qualitative study.

OBJECTIVES: Rheumatic heart disease (RHD) is a preventable disease frequently recognized in urban slums. Disease rates in Brazilian slums are incommensurate with the country's economic status and the existence of its universal healthcare system. Our study aimed to investigate what system issues may allow for disease persistence, focusing on issues surrounding access and utilization of primary and specialized healthcare services. STUDY DESIGN: This was a two-part (formative phase followed by implementation phase) qualitative study based on interviews and focus groups and analyzed via content analysis. METHODS: One focus group and 17 in-depth interviews with community health workers, primary care providers, and cardiologists who serve slum residents in Brazil and six interviews with key informants (community health researchers and cardiologists) were performed. Interviews with community health workers and primary care providers were from a single heath post in the neighborhood of Liberdade, a populous and previously unstudied slum in Salvador. Cardiologists were recruited from tertiary care referral hospitals in Salvador. RESULTS: Our findings revealed six major chronological categories/themes of issues and twenty subthemes that patients must overcome to avoid developing RHD or to have it successfully medically managed. Major themes include the effects of living in a slum (1), barriers to access and utilization of primary healthcare services (2), treatment in primary healthcare services (3), access/utilization of specialized healthcare services (4), treatment in specialized healthcare services (5), and certain systemic issues (6). CONCLUSION: Slums make residents sick in a manner of ways, and various bottlenecks impeding medical access to both primary care and specialty care exist, requiring multifaceted interventions. We detail major themes and finally suggest interventions that can allow for the health system to successfully eliminate RHD as a public health concern for slum residents.

Mycobacterium tuberculosis Mce1 protein complex initiates rapid induction of transcription of genes involved in substrate trafficking.

The mammalian cell entry (Mce)1 protein complex has an important role during the initial phase of a Mycobacterium tuberculosis (M. tuberculosis) infection. Murine macrophages were infected with M. tuberculosis H37Rv or Δ-mce1 H37Rv, and total RNA was isolated from the host cells at 15, 30 and 60 min, and 4 and 10 h post-infection. With the aim of studying the role for the Mce1 protein complex on host gene expression, the RNA was hybridized onto 44 K whole-genome microarrays. Selected genes were verified by reverse-transcriptase quantitative PCR (RT-QPCR). 'Transport' was the most overrepresented biological process during the first hour post H37Rv infection. Five genes (Abca1 (21.0-fold), Slc16a10 (3.1-fold), Slc6a12 (17.9-fold), Slc6a8 (2.3-fold) and Nr1h3, (5.5-fold)) involved in substrate trafficking were verified by RT-QPCR to be upregulated by >2-fold 1 h post H37Rv infection. By 1 h post Δ-mce1 H37Rv infection, only Abca1 and Slc6a12 were upregulated by >2-fold. A number of other genes, which may be directly involved in substrate trafficking or share the same transcription, were found to have expression profiles similar to the genes involved in substrate trafficking. The Mce1 protein complex has a significant role in the transcriptional activation of genes involved in substrate trafficking during the initial phase of an M. tuberculosis infection.

Severe infection in a lung transplant recipient caused by donor-transmitted carbapenem-resistant Acinetobacter baumannii.

We describe a case of proven donor transmission of carbapenem-resistant Acinetobacter baumannii, which resulted in severe infectious complications after lung transplantation. A single bla(OXA-23) positive strain, belonging to a new multilocus sequence type (ST231), was isolated from donor and recipient, who died 65 days after transplantation. This report highlights the current challenges associated with the potential transmission of multidrug-resistant infections through organ transplantation.

A novel antioxidant gene from Mycobacterium tuberculosis.

Among the major antimicrobial products of macrophages are reactive intermediates of the oxidation of nitrogen (RNI) and the reduction of oxygen (ROI). Selection of recombinants in acidified nitrite led to the cloning of a novel gene, noxR1, from a pathogenic clinical isolate of Mycobacterium tuberculosis. Expression of noxR1 conferred upon Escherichia coli and Mycobacterium smegmatis enhanced ability to resist RNI and ROI, whether the bacteria were exposed to exogenous compounds in medium or to endogenous products in macrophages. These studies provide the first identification of an RNI resistance mechanism in mycobacteria, point to a new mechanism for resistance to ROI, and raise the possibility that inhibition of the noxR1 pathway might enhance the ability of macrophages to control tuberculosis.

Cloning of an M. tuberculosis DNA fragment associated with entry and survival inside cells.

Mycobacterium tuberculosis infects one-third of the world's human population. This widespread infection depends on the organism's ability to escape host defenses by gaining entry and surviving inside the macrophage. DNA sequences of M. tuberculosis have been cloned; these confer on a nonpathogenic Escherichia coli strain an ability to invade HeLa cells, augment macrophage phagocytosis, and survive for at least 24 hours inside the human macrophage. This capacity to gain entry into mammalian cells and survive inside the macrophage was localized to two distinct loci on the cloned M. tuberculosis DNA fragment.

Reactive nitrogen intermediate susceptibility of Mycobacterium tuberculosis genotypes in an urban setting.

BACKGROUND: Mycobacterium tuberculosis genotypes resistant to reactive nitrogen intermediates (RNI) predominate in certain urban communities, suggesting that this phenotype influences disease transmission. OBJECTIVE: To compare different M. tuberculosis genotypes for resistance to RNI generated in vitro. DESIGN: We genotyped 420 M. tuberculosis isolates from a neighborhood in Sao Paulo, Brazil, and analyzed them for susceptibility to RNI generated in acidified sodium nitrite (ASN) solution. RESULTS: Seventy-one (43%) of 167 recent-infection strains and 68 (43%) of 158 endogenous infection strains showed moderate- to high-level ASN resistance. CONCLUSION: ASN resistance of M. tuberculosis is not necessarily a determining factor for enhanced transmission.

Risk factors for failure to complete a course of latent tuberculosis infection treatment in Salvador, Brazil.

BACKGROUND: Although treatment of latent tuberculosis infection (LTBI) is an essential component of tuberculosis (TB) control in countries such as the United States, it is not widely practiced in most TB-endemic countries. OBJECTIVE: To examine the practice of and adherence to LTBI treatment in a high-risk population in Brazil. DESIGN: We followed household contacts (HHCs) of patients hospitalized with pulmonary TB in Salvador, Brazil, for 6 months after they initiated LTBI treatment with isoniazid (INH). HHCs were asked to return to the hospital once a month for 6 months for follow-up visits and INH refills. RESULTS: Of 101 HHCs who initiated LTBI treatment, 54 (53.5%) completed the 6-month regimen. The risk of treatment non-completion was significantly higher in HHCs who reported side effects to INH (RR 2.69, 95%CI 1.3-5.8, P = 0.01), and in those who had to take two buses for a one-way trip to the hospital (RR 1.8, 95%CI 1.01-3.3, P = 0.04). Of the 101 HHCs, 29 (28.7%) did not return for any follow-up visits; these HHCs were significantly more likely to have a 2-bus commute to the hospital compared to HHCs who completed treatment (OR 20.69, 95%CI 2.1-208.4, P = 0.01). CONCLUSION: Nearly 50% of HHCs at high risk for developing TB completed a 6-month course of LTBI treatment. Completion of LTBI treatment was most affected by medication intolerance and commuting difficulties for follow-up visits.

Analysis of discordance between the tuberculin skin test and the interferon-gamma release assay.

OBJECTIVE: To analyze factors associated with discordance between tuberculin skin test (TST) and interferon-gamma release assay (IGRA) results among household contacts of pulmonary tuberculosis (PTB) patients. DESIGN: TST (purified protein derivative) and IGRA (QuantiFERON-TB Gold) were performed on household contacts of PTB patients diagnosed between 2006 and 2007 in Salvador, Brazil. Discordant test groups were compared with the TST-/IGRA- group. RESULTS: Of 261 household contacts satisfactorily tested by TST, 145 (55.6%) had positive TST results; of 298 satisfactorily tested by IGRA, 127 (43.1%) had positive results. The test agreement was 0.76 (kappa = 0.53, 95%CI 0.43-0.63). Sixty-one (24%) were discordant: 44 (72%) with TST+/IGRA- and 17 (28%) with TST-/IGRA+ results. Compared to the TST-/IGRA- group, the TST+/IGRA- and TST+/IGRA+ groups were significantly more likely to have a chest X-ray showing old lung scars (OR = 6.8, 95%CI 1.3-35.0; OR = 7.4, 95%CI 2.2-24.4, respectively). The TST-/IGRA+ group was exposed to their index cases for significantly longer than the TST-/IGRA- group (OR = 7.2, 95%CI 1.7-29.3). CONCLUSION: The TST+/IGRA- and TST+/IGRA+ groups shared more similar characteristics with each other than with the TST-/IGRA- group. In a setting endemic for TB, TST results appear to be more suitable in the decision to treat latent TB infection.

A multiplexed, indirect enzyme-linked immunoassay for the detection and differentiation of E. coli from other Enterobacteriaceae and P. aeruginosa from other glucose non-fermenters.

Gram-negative bacteria (GNB) are important causes of community (CA) and hospital (HA)- associated infections. Here we describe the development of an indirect ELISA (I-ELISA), which can be used to detect and differentiate the Enterobacteriaceae Escherichia coli, and glucose non-fermenter Pseudomonas aeruginosa from other GNB species. The I-ELISA utilizes six antibodies for bacterial speciation, which were grouped according to their bacterial targets; Enterobacteriaceae (SL-EntA and CH1810 mAb), Escherichia coli (SL-EcA and 6103-46 mAb), Pseudomonas aeruginosa (SL-PaA and SL-PaB). The six, anti-GNB antibodies were first screened against a panel of well-characterized clinical GNB isolates to optimize assay conditions and to determine individual antibody sensitivity and specificity. When tested against a diverse, blinded panel of 94 GNB clinical isolates, the I-ELISA exhibited the following sensitivity/specificity for each target: Enterobacteriaceae (94.4%/95%), E. coli (82.6%/88.7%), P. aeruginosa (83.3%/96%). An I-ELISA to detect and differentiate the most common GNB pathogens offers advantage in terms of simplicity over diagnostic tests currently used in most clinical settings.

Epidemiology of meningococcal disease in southern Brazil from 1995 to 2003, and molecular characterization of Neisseria meningitidis using multilocus sequence typing.

OBJECTIVE: To describe the epidemiology of meningococcal disease (MD) in southern Brazil. METHODS: Retrospective cohort study among 2215 MD cases reported from 1995 to 2003 in Rio Grande do Sul (RS) State. RESULTS: The overall incidence fell by 50%; the case-fatality rate during this period was 22%. Even so, the incidence of MD remained high after the epidemic period ended in 1999. Together, the age groups of 1-4 years and infants accounted for 54.1% of reported cases with incidences of 11.3/100 000 and 31.3/100 000, respectively; 69.8% of cases were caused by Neisseria meningitidis serogroup B, which increased significantly. There was a significant decrease in serogroup C cases in the whole period. The phenotypes B:4,7:P1.19,15, B:15:P1.7,16 and B:NT:P1.3 caused almost 50% of all serotyped cases. Fifty-six isolates obtained from RS patients during the first non-epidemic year 2000 plus 20 isolates from other southern Brazilian states (Santa Catarina and Paraná), Denmark and France were typed by multilocus sequence typing. Twenty sequence types (STs) were identified, eight of them found only in RS. ST-33 (27%) and ST-259 (18%) were the most frequent; both belong to the ST-32/ET-5 complex. ST-259 cases showed a trend towards higher risk of fatal outcome. ST-259 isolates were not detected among geographic controls or in other studies in Brazil. CONCLUSION: Our data suggest that ST-33 and ST-259 clones and the emergence of the ST-103 isolates contributed to the continued high incidence of MD in RS.

Serum antiphospholipid antibody levels as biomarkers for diagnosis of pulmonary tuberculosis patients.

SETTING: Salvador, Bahia, Brazil. OBJECTIVE: To evaluate the immunoglobulin (Ig)M and total IgG antibody response to cardiolipin (CL), phosphatidylcholine (PTC), phosphatidylethanolamine (PE), phosphatidylinositol (PI) and sulfatide (SL-I) as biosignatures that can be used to diagnose pulmonary tuberculosis (TB) and its applicability for monitoring the efficacy of anti-tuberculosis treatment. DESIGN: Serum samples from 37 adult pulmonary TB patients and 48 controls (16 healthy household contacts, 19 household contacts with latent tuberculous infection [LTBI] and 13 non-TB patients with lung disease) were screened using enzyme-linked immunosorbent assays (ELISAs) for IgM and total IgG against phospholipids. RESULTS: Levels of IgM response to CL, PE and PI, and IgG response to CL, PE, PI and PTC were significantly higher in TB patients than in control groups. Anti-CL IgG had the best performance characteristics, with a sensitivity and specificity of respectively 86.5% and 87.2%. This IgG anti-CL ELISA test detected 86.5% (32/37) of the TB patients, whereas the number detected using sputum smear was only 65.9% (24/37). After anti-tuberculosis treatment, the median value for all anti-phospholipid antibodies decreased significantly compared with baseline values (P < 0.05). CONCLUSION: Our results suggest that the total IgG anti-CL level could be useful to complement conventional bacteriological tests for the rapid diagnosis of adult pulmonary TB.

A case of severe pancreatitis complicated by Raoultella planticola infection.

A 45-year-old-male presented with severe pancreatitis. Two bacterial isolates obtained from peritoneal fluid and abdominal purulent secretion were identified to the species level by 15 biochemical tests and four supplementary tests as Raoultella planticola. Identification was confirmed by rpoB gene sequencing. R. planticola is difficult to identify in the clinical laboratory, and the clinical significance of this isolation remains uncharacterized. This is the first report of pancreatitis with a primary infection by R. planticola.

Diabetes increases the risk of recent-transmission tuberculosis in household contacts in São Paulo, Brazil.

SETTING: A cohort of household contacts of tuberculosis (TB) index cases from four public health clinics in São Paulo, Brazil. OBJECTIVE: To measure the association between diabetes mellitus (DM) among household contacts and recent-transmission TB (RT TB). DESIGN: Index TB cases (n = 263) identified from 2001 to 2002 in São Paulo, whose household contacts (n = 1383) were monitored for active TB until December 2010. RESULTS: From 2001 to 2010, there were 29 cases of RT TB among household contacts (cumulative incidence 2.1%, 95%CI 1.4-2.9). DM in household contacts was associated with RT TB (OR 3.96, 95%CI 1.33-11.79) even after adjustment for human immunodeficiency virus (HIV) status, smoking and alcohol use (adjusted OR [aOR] 3.21, 95%CI 1.01-10.19). HIV infection was also associated with RT TB (OR 6.40, 95%CI 1.40-29.40; aOR 4.81, 95%CI 0.96-24.18). Household contact DM was not associated with non-RT TB (OR 1.27, 95%CI 0.30-5.40). The time to diagnosis of TB was shorter in household contacts with and without DM (P = 0.035) and in household contacts with and without HIV (P = 0.0002). CONCLUSION: Household contact DM was associated with an increased risk of RT TB in a cohort in Brazil, lending support to the active screening of household contacts with DM for TB in Brazil.

Pseudomonas aeruginosa epidemic strain carrying bla(SPM) metallo-beta-lactamase detected in Rio de Janeiro, Brazil.

The present study was designed to characterize beta-lactamase genes and evaluate polymerase chain reaction (PCR) typing for multidrug-resistant Pseudomonas aeruginosa pulsed-field gel electrophoresis (PFGE) genotype A isolates from Rio de Janeiro, Brazil, collected between April 1999 and March 2000 and one additional isolate collected in June 2002. As reported previously, all of the genotype A isolates produced non-characterized metallo-beta-lactamase. These isolates (22) were screened for the bla(SPM) gene by PCR and dot-blotting. Isolates were typed by PCR fingerprinting with primers RAPD-1, 272, 208, 1290, ERIC-1 and ERIC-2. The bla(SPM) gene was detected in 18 (82%) of the 22 isolates. PCR fingerprinting gave results that correlated with PFGE, except with primer 1290. In Rio de Janeiro and other Brazilian states, nearly all SPM-producing P. aeruginosa isolates belong to a single PFGE type accounting for a large proportion of drug-resistant P. aeruginosa hospital infections. RAPD PCR fingerprinting may be a useful technique to screen for an epidemic multidrug-resistant strain in Brazil.

Structure and functions of hepatitis C virus proteins: 15 years after.

Since its discovery in 1988, the hepatitis C virus (HCV) has become a hot topic of research by many groups around the world. This globally spread infectious agent is responsible for a large proportion of chronic viral hepatitides. The clue to halting the hepatitis C pandemic may be the detailed understanding of the virus structure, its replication mechanism, and the exact functions of the various proteins. Such understanding could enable the development of new antivirals targeted against hepatitis C virus and possibly an effective vaccine. This review recaps the current knowledge about the HCV genome 15 years after its discovery. The structure and function of particular viral structural (core, E1, E2) and nonstructural (NS2, NS3, NS4, NS5) proteins and noncoding regions known to date are described. With respect to frequent conflicting reports from different research groups, results reproducibly demonstrated by independent investigators are emphasized. Owing to many obstacles and limitations inherent in doing research on this noteworthy virus, the current knowledge is incomplete and the answers to many important questions are to be expected in the future.

Determinants of cell entry and intracellular survival of Mycobacterium tuberculosis.

The ability of Mycobacterium tuberculosis to sustain a chronic infection and cause disease in a subset of those infected depends on its products--virulence factors--that enable the organism to enter and survive indefinitely inside mononuclear phagocytic cells by subverting cellular antimicrobial mechanisms. Characterizing these factors is essential to understanding the pathogenesis of M. tuberculosis.

A population-based study of drug resistance and transmission of tuberculosis in an urban community.

SETTING: A low-income neighborhood of Sao Paulo, Brazil. OBJECTIVE: To determine the incidence, risk factors and transmission patterns of multidrug-resistant tuberculosis (MDR-TB). DESIGN: Prospective longitudinal study of patients with pulmonary TB (PTB). METHODS: Sputum culture-confirmed patients with PTB were recruited between March 2000 and May 2002. Bivariate and multivariate logistic regression analyses were performed to identify factors associated with MDR-TB. Mycobacterium tuberculosis isolates were tested for drug susceptibility and typed by IS6110-RFLP analysis. RESULTS: Of 420 patients, respectively 71% and 27% were new and previously treated; 15.5% of the patients' M. tuberculosis isolates were resistant to at least one drug; of these, 11% and 27% were found among new and previously treated cases, respectively. Respectively 1% and 16.7% of the new and previously treated cases were MDR-TB. RFLP analysis showed that new transmission of MDR strains was uncommon. By multivariate logistic regression analysis, previous TB and hospitalization in the 24 months before TB diagnosis were identified as independent predictors of MDR-TB. CONCLUSIONS: The results showed an intermediate level of MDR-TB incidence in a neighborhood of Sao Paulo and identified predictors that can be targeted for intervention by national and local TB control programs.

Campylobacter infections in the United States. Results of an 11-state surveillance.

In January 1982, 11 states (Alabama, Arizona, Georgia, Illinois, Minnesota, New Mexico, Oregon, Texas, Vermont, Washington, and Wisconsin) began reporting monthly their isolations of Campylobacter to the Centers for Disease Control in Atlanta. The information reported included the species of Campylobacter organisms, the week of the report, the site from which the organism was isolated, and the age and sex of the infected person. A total of 3,966 isolates were reported in 1982, of which 3,900 were Campylobacter jejuni. Campylobacter isolations exceeded Salmonella in two of the three states (Oregon and Wisconsin) that require reporting. The eight other states with lower rates of isolation had variable reporting practices. Rates of Campylobacter isolations were highest in June through August. Age-specific rates of Campylobacter infections peaked in the 1- to 2-year and 20- to 29-year age groups. Fifty-five percent of all isolates were from male patients. Campylobacter infections seem to be at least as common as Salmonella infections in states in which the reporting practices are comparable.

Directly observed therapy of tuberculosis in Brazil: associated determinants and impact on treatment outcome.

SETTING: All Brazilian states. OBJECTIVES: To assess the determinants of tuberculosis (TB) in patients undergoing directly observed therapy (DOT) and the impact of DOT on treatment outcomes. DESIGN: This is a cross-sectional study among TB patients aged ⩾18 years conducted in 2011. The primary outcome was the status of DOT received, while the secondary was the outcome of anti-tuberculosis treatment. RESULTS: In 2011, 35 775 (38.3%) subjects received DOT. The odds of receiving DOT were higher in patients with the following characteristics: brown/mestizo patients (OR 1.18, 95%CI 1.14-1.22) and those of other ethnic groups (OR 2.01, 95%CI 1.79-2.27) compared to Whites, alcohol users (OR 1.37, 95%CI 1.28-1.47) and those with mental disorders (OR 1.88, 95%CI 1.54-2.29). The odds of receiving DOT were lower in human immunodeficiency virus positive patients (OR 0.64, 95%CI 0.60-0.68). Patients who did not receive DOT were more likely to default from anti-tuberculosis treatment (OR 0.62, 95%CI 0.57-0.66), die due to TB (OR 0.68, 95%CI 0.61-0.77) and to have unknown treatment outcomes (OR 0.71, 95%CI 0.66-0.76). The adjusted preventable fraction of DOT in the reduction of unfavorable outcomes was 25%. CONCLUSION: Sociodemographic and clinical characteristics are determinants of anti-tuberculosis treatment outcomes in patients undergoing DOT; DOT use led to a 25% reduction in unfavorable outcomes.

Energy transfer between fluorescent proteins using a co-expression system in Mycobacterium smegmatis.

The goal of this study was to establish a two-plasmid co-expression system for Mycobacterium smegmatis. Two vectors with compatible origins of replication and a polylinker, which allows modular cloning of promoters and genes, were constructed and used to clone genes encoding a blue fluorescent protein (BFP) and a green fluorescent protein (GFP). A 160-fold variation of GFP expression levels in M. smegmatis was achieved by combining three promoters with different copy numbers of the vectors. An efficient energy transfer between BFP and GFP in M. smegmatis was observed by fluorescence measurements and demonstrated that these genes were simultaneously expressed from both vectors. Thus, these vectors will be valuable for all strategies where co-expression of proteins in M. smegmatis is needed, e.g. for constructing a two-hybrid system or for deleting essential genes.