A retrospective evaluation of a score system adopted by the Ministry of Health, Brazil in the diagnosis of pulmonary tuberculosis in childhood: a case control study

Based on a retrospective case-control study we evaluated the score system adopted by the Ministry of Health of Brazil (Ministério da Saúde - MS), to diagnose pulmonary tuberculosis (PTB) in childhood. This system is independent of bacteriological or histopathological data to define a very likely (> or = 40 points), possible (30-35 points) or unlikely (< or = 25 points) diagnosis of tuberculosis. Records of hospitalized non-infected HIV children at the Instituto de Puericultura e Pediatria Martagão Gesteira of Federal University of Rio de Janeiro (IPPMG-UFRJ), were reviewed. Patients were adjusted for age and divided in two different groups: 45 subjects in the case group (culture-positive) [mean of age = 10.64 mo; SD 9.66]; and 96 in the control group (culture-negative and clinic criteria that dismissed the disease) [mean of age = 11.79 mo.; SD 11.31]. Among the variables analyzed, the radiological status had the greater impact into the diagnosis (OR = 25.39), followed by exposure to adult with tuberculosis (OR = 10.67), tuberculin skin test >10mm (OR = 8.23). The best cut-off point to the diagnosis of PTB was 30 points, where the score system was more accurate, with sensitivity of 88.9% and specificity of 86.5%.

QUANTITATIVE REAL-TIME PCR (Q-PCR) FOR SPUTUM SMEAR DIAGNOSIS OF PULMONARY TUBERCULOSIS AMONG PEOPLE WITH HIV/AIDS

Objective: To assess quantitative real-time polymerase chain reaction (q-PCR) for the sputum smear diagnosis of pulmonary tuberculosis (PTB) in patients living with HIV/AIDS with a clinical suspicion of PTB.Method: This is a prospective study to assess the accuracy of a diagnostic test, conducted on 140 sputum specimens from 140 patients living with HIV/AIDS with a clinical suspicion of PTB, attended at two referral hospitals for people living with HIV/AIDS in the city of Recife, Pernambuco, Brazil. A Löwenstein-Jensen medium culture and 7H9 broth were used as gold standard.Results: Of the 140 sputum samples, 47 (33.6%) were positive with the gold standard. q-PCR was positive in 42 (30%) of the 140 patients. Only one (0.71%) did not correspond to the culture. The sensitivity, specificity and accuracy of the q-PCR were 87.2%, 98.9% and 95% respectively. In 39 (93%) of the 42 q-PCR positive cases, the CT (threshold cycle) was equal to or less than 37.Conclusion: q-PCR performed on sputum smears from patients living with HIV/AIDS demonstrated satisfactory sensitivity, specificity and accuracy, and may therefore be recommended as a method for diagnosing PTB.

PRETERM BIRTH AND FETAL GROWTH RESTRICTION IN HIV-INFECTED BRAZILIAN PREGNANT WOMEN

Introduction: Maternal HIV infection and related co-morbidities may have two outstanding consequences to fetal health: mother-to-child transmission (MTCT) and adverse perinatal outcomes. After Brazilian success in reducing MTCT, the attention must now be diverted to the potentially increased risk for preterm birth (PTB) and intrauterine fetal growth restriction (IUGR). Objective: To determine the prevalence of PTB and IUGR in low income, antiretroviral users, publicly assisted, HIV-infected women and to verify its relation to the HIV infection stage. Patients and Methods: Out of 250 deliveries from HIV-infected mothers that delivered at a tertiary public university hospital in the city of Vitória, state of Espírito Santo, Southeastern Brazil, from November 2001 to May 2012, 74 single pregnancies were selected for study, with ultrasound validated gestational age (GA) and data on birth dimensions: fetal weight (FW), birth length (BL), head and abdominal circumferences (HC, AC). The data were extracted from clinical and pathological records, and the outcomes summarized as proportions of preterm birth (PTB, < 37 weeks), low birth weight (LBW, < 2500g) and small (SGA), adequate (AGA) and large (LGA) for GA, defined as having a value below, between or beyond the ±1.28 z/GA score, the usual clinical cut-off to demarcate the 10th and 90th percentiles. Results: PTB was observed in 17.5%, LBW in 20.2% and SGA FW, BL, HC and AC in 16.2%, 19.1%, 13.8%, and 17.4% respectively. The proportions in HIV-only and AIDS cases were: PTB: 5.9 versus 27.5%, LBW: 14.7% versus 25.0%, SGA BW: 17.6% versus 15.0%, BL: 6.0% versus 30.0%, HC: 9.0% versus 17.9%, and AC: 13.3% versus 21.2%; only SGA BL attained a significant difference. Out of 15 cases of LBW, eight (53.3%) were preterm only, four (26.7%) were SGA only, and three (20.0%) were both PTB and SGA cases. A concomitant presence of, at least, two SGA dimensions in the same fetus was frequent. Conclusions: The proportions of preterm birth and low birth weight were higher than the local and Brazilian prevalence and a trend was observed for higher proportions of SGA fetal dimensions than the expected population distribution in this small casuistry of newborn from the HIV-infected, low income, antiretroviral users, and publicly assisted pregnant women. A trend for higher prevalence of PTB, LBW and SGA fetal dimensions was also observed in infants born to mothers with AIDS compared to HIV-infected mothers without AIDS.

Colorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detection

Direct smear examination using Ziehl-Neelsen staining for pulmonary tuberculosis (PTB) diagnosis is inexpensive and easy to use, but has the major limitation of low sensitivity. Rapid molecular methods are becoming more widely available in centralized laboratories, but they depend on timely reporting of results and strict quality assurance obtainable only from costly commercial kits available in high burden nations. This study describes a pre-commercial colorimetric method, Detect-TB, for detecting Mycobacterium tuberculosis DNA in which an oligonucleotide probe is fixed onto wells of microwell plates and hybridized with biotinylated polymerase chain reaction amplification products derived from clinical samples. The probe is capable of hybridising with the IS6110 insertion element and was used to specifically recognise the M. tuberculosis complex. When combined with an improved silica-based DNA extraction method, the sensitivity of the test was 50 colony-forming units of the M. tuberculosis reference strain H37Rv. The results that were in agreement with reference detection methods were observed in 95.2% (453/476) of samples included in the analysis. Sensitivity and specificity for 301 induced sputum samples and 175 spontaneous sputum samples were 85% and 98%, and 94% and 100%, respectively. This colorimetric method showed similar specificity to that described for commercially available kits and may provide an important contribution for PTB diagnosis.

Immunologic evaluation and validation of methods using synthetic peptides derived from Mycobacterium tuberculosis for the diagnosis of tuberculosis infection

The goal of this study was to demonstrate the usefulness of an enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of pulmonary tuberculosis (PTB) and extrapulmonary TB (EPTB). This assay used 20 amino acid-long, non-overlapped synthetic peptides that spanned the complete Mycobacterium tuberculosis ESAT-6 and Ag85A sequences. The validation cohort consisted of 1,102 individuals who were grouped into the following five diagnostic groups: 455 patients with PTB, 60 patients with EPTB, 40 individuals with non-EPTB, 33 individuals with leprosy and 514 healthy controls. For the PTB group, two ESAT-6 peptides (12033 and 12034) had the highest sensitivity levels of 96.9% and 96.2%, respectively, and an Ag85A-peptide (29878) was the most specific (97.4%) in the PTB groups. For the EPTB group, two Ag85A peptides (11005 and 11006) were observed to have a sensitivity of 98.3% and an Ag85A-peptide (29878) was also the most specific (96.4%). When combinations of peptides were used, such as 12033 and 12034 or 11005 and 11006, 99.5% and 100% sensitivities in the PTB and EPTB groups were observed, respectively. In conclusion, for a cohort that consists entirely of individuals from Venezuela, a multi-antigen immunoassay using highly sensitive ESAT-6 and Ag85A peptides alone and in combination could be used to more rapidly diagnose PTB and EPTB infection.

Correlations between major risk factors and closely related Mycobacterium tuberculosis isolates grouped by three current enotyping procedures: a population-based study in northeast Mexico

The characteristics of tuberculosis (TB) patients related to a chain of recent TB transmissions were investigated. Mycobacterium tuberculosis (MTB) isolates (120) were genotyped using the restriction fragment length polymorphism-IS6110 (R), spacer oligotyping (S) and mycobacterial interspersed repetitive units-variable number of tandem repeats (M) methods. The MTB isolates were clustered and the clusters were grouped according to the similarities of their genotypes. Spearman’s rank correlation coefficients between the groups of MTB isolates with similar genotypes and those patient characteristics indicating a risk for a pulmonary TB (PTB) chain transmission were ana- lysed. The isolates showing similar genotypes were distributed as follows: SMR (5%), SM (12.5%), SR (1.67%), MR (0%), S (46.67%), M (5%) and R (0%). The remaining 35 cases were orphans. SMR exhibited a significant correlation (p < 0.05) with visits to clinics, municipalities and comorbidities (primarily diabetes mellitus). S correlated with drug consumption and M with comorbidities. SMR is needed to identify a social network in metropolitan areas for PTB transmission and S and M are able to detect risk factors as secondary components of a transmission chain of TB.

Distribuição temporal de imaturos de Odonata (Insecta) associados a Eichhornia azurea (Kunth) na Lagoa do Camargo, Rio Paranapanema, São Paulo

Os objetivos deste trabalho foram registrar a abundância e a riqueza de Odonata associada a Eichhornia azurea, durante o período de março de 2004 a março de 2005, na Lagoa do Camargo, lateral ao Rio Paranapanema - São Paulo, após um pulso de inundação extraordinário e também investigar os fatores ambientais determinantes na distribuição da abundância de Odonata. As maiores abundâncias e riquezas ocorreram na estação seca, sendo que Coenagrionidae foi a família mais abundante e com a maior riqueza de gêneros de todo o período estudado. Esta alta abundância possivelmente ocorreu devido a seu comportamento, como postura dos ovos dentro do tecido das macrófitas e hábito escalador. Aeshnidae e Libellulidae apresentaram baixa abundância principalmente na estação seca. Os principais fatores ambientais que afetaram a distribuição da abundância de Odonata foram a temperatura de superfície da água, a pluviosidade e a biomassa de E. azurea.

Teores de cobre, zinco e ferro no fígado de búfalos (Bubalus bubalis) com paratuberculose

Resumo: Com o objetivo de realizar um estudo dos teores de cobre (Cu), zinco (Zn) e ferro (Fe) em búfalas com paratuberculose (PTB) foram utilizadas 13 búfalas, das raças Murrah, Mediterrâneo e seus mestiços acima de três anos de idade, pertencentes a duas propriedades localizadas nos municípios de São Luiz e São Mateus, no Estado do Maranhão. Os animais foram selecionados de acordo com a presença de sinais clínicos sugestivos de paratuberculose, caracterizados por estado nutricional regular a ruim, diarreia crônica líquida a semi-líquida, desidratação, edema submandibular, anestro prolongado, mastites e verminose gastrintestinal. Foi realizada biópsia retal em todos os animais, para detecção de Mycobacterium avium subsp. paratuberculosis (Map) por meio da qPCR, e exames histopatológicos (HE e Ziehl-Neelsen). No Grupo1 sete animais foram positivos para presença de Map, e no Grupo 2 seis foram negativos. Todos os búfalos foram eutanasiados e necropsiados para coleta de diversos tecidos. Parte dos fragmentos foram fixados em formol a 10% para histopatologia e fragmentos de tecido hepático foram congelados para as dosagens dos microminerais (Cu, Zn e Fe). À necropsia todos os animais positivos para PTB apresentavam linfonodos mesentéricos de coloração castanha sugestiva de hemossiderose. Adicionalmente, em um animal foram observados pequenos pontos de cor marrom distribuídos difusamente na mucosa do intestino delgado. Na histopatologia foi observada hemossiderose moderada a acentuada no baço dos animais do Grupo 1. Na dosagem dos microminerais todos os animais com PTB apresentaram níveis abaixo dos valores de referência para Cu e Zn. Observou-se que a média dos teores de Cu dos búfalos com PTB foi 18,0ppm e de Zn 68,6ppm. No Grupo 2 a média dos teores de Cu foi 113,7ppm e de Zn 110,0ppm. Os teores de Fe em ambos os grupos foram elevados (>670ppm). Baseado nos achados clínico-patológicos e nas dosagens de minerais realizadas neste estudo, conclui-se que na região estudada, a PTB agravou o quadro clínico de animais com deficiência de Cu e Zn. Em áreas menos deficientes desses minerais sugere-se que a doença seja capaz de induzir quadros de deficiência mineral secundária.

Fitoplâncton em três lagoas marginais ao Rio Paranapanema e em sua desembocadura no Reservatório de Jurumirim (São Paulo, Brasil) durante um período prolongado de seca

Durante um período de desconexão (de outubro de 1999 a dezembro de 2000) entre lagoas laterais e o Rio Paranapanema, a estrutura da comunidade fitoplanctônica e suas alterações foram examinadas em amostras coletadas trimestralmente nas zonas litoral e pelágica. Um total de 183 táxons de algas foi encontrado, sendo a comunidade predominante constituída por Chlorophyceae e Bacillariophyceae no Rio Paranapanema e em um ambiente lacustre permanentemente isolado, e por Bacillariophyceae e Chlorophyceae em duas outras lagoas. Ao longo do ano, diminuição na riqueza de espécies foi encontrada somente no ambiente lêntico com maior perda de conectividade. Cryptophyceae foi abundante (densidades maior que a densidade média do fitoplâncton) nas três lagoas, zonas e épocas do ano, exceto na zona litoral da lagoa com maior perda de conectividade em novembro de 1999. No Rio Paranapanema, Cryptophyceae predominou em mês chuvoso (fevereiro), sendo os índices de diversidade mais elevados nos demais períodos. A seca severa acarretou redução significativa do volume e área de superfície das lagoas, uma delas, a lagoa isolada, sendo extinta no último período de amostragem. Somente, nesta última, um efeito de concentração na abundância do fitoplâncton foi evidente. O predomínio em densidade de Cryptomonas brasiliensis Castro, C. Bic. & D. Bic., Chroomonas spp. e Chlamydomonas spp. nas três lagoas foi atribuído às condições adversas para o desenvolvimento das demais algas. A baixa profundidade, a grande quantidade de sedimentos suspensos na água após episódios de ventos fortes e, em conseqüência, a extinção acentuada da luz, foram fatores limitantes para as algas não adaptadas a essas condições, a despeito das concentrações favoráveis de nutrientes. No Rio Paranapanema, as alterações temporais na estrutura da comunidade fitoplanctônica parecem estar associadas a flutuações de velocidade e fluxo de água bem como ao nível hidrológico.

Social inequality and perinatal health: comparison of three Brazilian cohorts

The objective of the present study was to estimate and compare social inequality in terms of three indicators, i.e., low birth weight (LBW), preterm birth (PTB) and small for gestational age (SGA) birth, in three birth cohorts. Two cohorts were from the city of Ribeirão Preto, where data were collected for all 6748 live born singletons in 1978/79 and for one third of live born singletons (2846) in 1994. The third cohort consisted of 2443 singletons born in São Luís over a period of one year (1997/98). In Ribeirão Preto, LBW and PTB rates increased in all social strata from 1978/79 to 1994. Social inequalities regarding LBW and PTB disappeared since the increase in these rates was more accelerated in the groups with higher educational level. The percentage of SGA infants increased over the study period. Social inequality regarding SGA birth increased due to a more intense increase in SGA births in the strata with lower schooling. In São Luís, in 1997/98 there was no social inequality in LBW or PTB rates, whereas SGA birth rate was higher in mothers with less schooling. We speculate that the more accelerated increase in medical intervention, especially due to the increase in cesarean sections in the more privileged groups, could be the main factor explaining the unexpected increase in LBW and PTB rates in Ribeirão Preto and the decrease or disappearance of social inequality regarding these perinatal indicators in the two cities.