Diagnosis of hepatitis C virus in Brazilian blood donors using a reverse transcriptase nested polymerase chain reaction: comparison with enzyme immunoassay and recombinant protein immunoblot assay

Screening blood donations for anti-HCV antibodies and alanine aminotransferase (ALT) serum levels generally prevents the transmission of hepatitis C virus (HCV) by transfusion. The aim of the present study was to evaluate the efficiency of the enzyme immunoassay (EIA) screening policy in identifying potentially infectious blood donors capable to transmit hepatitis C through blood transfusion. We have used a reverse transcriptase (RT)-nested polymerase chain reaction (PCR) to investigate the presence of HCV-RNA in blood donors. The prevalence of HCV-RNA positive individuals was compared with the recombinant immunoblot assay (RIBA-2) results in order to assess the usefulness of both tests as confirmatory assays. Both tests results were also compared with the EIA-2 OD/C ratio (optical densities of the samples divided by the cut off value). ALT results were expressed as the ALT quotient (qALT), calculated dividing the ALT value of the samples by the maximum normal value (53UI/l) for the method. Donors (n=178) were divided into five groups according to their EIA anti-HCV status and qALT: group A (EIA > or = 3, ALT<1), group B (EIA > or = 3, ALT>1), group C (1<=EIA<3, ALT<1), group D (1<=EIA<3, ALT>1) and group E (EIA<=0.7). HCV sequences were detected by RT-nested PCR, using primers for the most conserved region of viral genome. RIBA-2 was applied to the same samples. In group A (n=6), all samples were positive by RT-nested PCR and RIBA-2. Among 124 samples in group B, 120 (96.8%) were RIBA-2 positive and 4 (3.2%) were RIBA-2 indeterminate but were seropositive for antigen c22.3. In group B, 109 (87.9%) of the RIBA-2 positive samples were also RT-nested PCR positive, as well as were all RIBA-2 indeterminate samples. In group C, all samples (n=9) were RT-nested PCR negative: 4 (44.4%) were also RIBA-2 negative, 4 (44.4%) were RIBA-2 positive and 1 (11.1%) was RIBA-2 indeterminate. HCV-RNA was detected by RT-nested PCR in 3 (37.5%) out of 8 samples in group D. Only one of them was also RIBA-2 positive, all the others were RIBA-2 indeterminate. All of the group E samples (controls) were RT- nested PCR and RIBA-2 negative. Our study suggests a strong relation between anti-HCV EIA-2 ratio > or = 3 and detectable HCV-RNA by RT-nested PCR. We have also noted that blood donors with RIBA-2 indeterminate presented a high degree of detectable HCV-RNA using RT-nested PCR (75%), especially when the c22.3 band was detected.

Functional and structural characterization of the RNase II-family of enzymes

Dissertation presented to obtain a Doctoral Degree in Biology by Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa

The association between serum potassium at hospital admission and the case-fatality rate of leptospirosis in men

Severe leptospirosis affects predominantly males and presents a high susceptibility to hypokalemic acute renal failure. As hypokalemia and hyperkalemia induce severe complications, it is important to evaluate if the initial serum potassium is an independent risk factor for death in leptospirosis. The medical records of 1016 patients hospitalized with the diagnosis of leptospirosis were reviewed. The analysis was restricted to 442, according to the following criteria: male, 18 years or older, information about death or hospital discharge and recorded values of serum potassium, serum creatinine and duration of symptoms at admission. Potassium values lower than 3.5 mEq/L (hypoK), 3.5-5 mEq/L (normoK) and above 5 mEq/L (hyperK) were detected in 180, 245 and 17 patients, respectively. The death rate increased with serum potassium: 11.1% in the hypoK, 14.7% in the normoK and 47.1% in the hyperK group (p = 0.002). In a logistic regression model (normoK as referent), including age, creatinine and duration of symptoms, hypoK was not associated with increased death rate (odds ratio (OR) = 0.80; p > 0.1). On the other hand, hyperK showed a significant association with increased risk of death (OR = 3.95, p = 0.021). In conclusion, in this sample of men with leptospirosis initial serum potassium was positively and independently correlated with the risk of in-hospital death.

Comparative study of the in-hospital case-fatality rate of leptospirosis between pediatric and adult patients of different age groups

The main objective was to compare the in-hospital case-fatality rate of leptospirosis between pediatric (< 19 years) and adult (>19 years) patients, taking into account gender, renal function, duration of symptoms and jaundice. Medical records of 1016 patients were reviewed. Comparative analysis was restricted to 840 patients (100 pediatric, 740 adults) with recorded information on the variables included in the analysis. Among these patients 81.7% were male and 91.5% were icteric. The case-fatality rate of leptospirosis was 14.4%. The odds of death adjusted for gender, jaundice, duration of symptoms, serum urea and serum creatinine were almost four times higher for the adult than for the pediatric group (odds ratio (OR) = 3.94; 95% confidence interval = 1.19-13.03, p = 0.029). Among adults, increased age was also significantly and independently associated with increased risk of death (p < 0.01). Older patients were also more often treated by dialysis. In conclusion, the data suggest that the in-hospital case fatality rate of leptospirosis is higher for adults than for children and adolescents, even after taking into account the effects of several potential risk factors of death. Among adults, older age was also strongly and independently associated with higher risk of death.

Fascioliasis in relatives of patients with Fasciola hepatica infection in Peru

High prevalence rates of human fascioliasis have been described in several regions of Peru. We surveyed 20 families in an endemic area of Peru in order to determine the proportion of infection with F. hepatica in relatives of diagnosed subjects and in order to identify associated risk factors. The study included feces and blood samples of 93 subjects. Ages ranged from one to 53 (mean = 18.6; SD = 14.2). The overall prevalence of fascioliasis by fecal examinations was 33.3% (n = 83) and by serology, 51.9% (n = 86). The prevalence in age group I (< 19 years old) by coprological and serological tests was 61.4% and 75.9%, respectively; in group II (> 19 years old) 15.4% and 37.5%. The main associated risk factor with fascioliasis was eating salads (OR = 3.29, CI = 1.2 - 9.0, p = 0.02). In conclusion, human fascioliasis is highly prevalent in the relatives of index cases and the most significant risk factor of acquiring fascioliasis in the family is eating salads in endemic areas.

Serological markers and risk factors for hepatitis B and C viruses in patients infected with human immunodeficiency virus

Both hepatitis B and hepatitis C viruses (HBV and HCV) infection are common in HIV-infected individuals as a result of shared risk factors for acquisition. A serological study for HBV and HCV was performed in 251 HIV-positive individuals from Medellín, Colombia. A qualitative RT-PCR for HCV was done in 90 patients with CD4+ T-cell count < 150 per mm³. Serological markers for HBV infection were present in 97 (38.6%) patients. Thirty six of them (37.1%) had isolated anti-HBc. A multivariate analysis indicated that the following risk factors were significantly associated with the presence of these markers: age (OR = 1.05, 95% CI: 1.01-1.08), pediculosis pubis (OR = 1.83, 95% CI: 1.01-3.33), men who have sex with men and women (OR = 3.23, 95% CI: 1.46-7.13) and men who have sex only with men (OR = 3.73, 95% CI: 1.58-8.78). The same analysis restricted to women showed syphilis as the only significant risk factor. Thus, HBV infection was considerably associated with high risk sexual behavior. HCV was present in only two (0.8%) of HIV patients. Both of them were positive by RT-PCR and anti-HCV. This low frequency of HIV/HCV coinfection was probably due to the uncommon intravenous drug abuse in this population. The frequent finding of isolated anti-HBc warrants molecular approaches to rule out the presence of cryptic HBV infection.

Risk Factors for Latex Sensitization in Children with Spina Bifida

Background: Children with spina bifida represent the major risk group for latex sensitization. Purpose: To determine the prevalence of latex sensitization in these children and to identify risk factors. Material and methods: We studied 57 patients with spina bifida. The mean age was 5.6 years and the male/female ratio was 0.8/1. In all patients a questionnaire, skin prick test (SPT) with latex (UCBStallergènes, Lofarma and ALK-Abelló), common aeroallergens and fruits (UCB-Stallergènes) and serum determination of total IgE (AlaSTAT) were performed. Results: The prevalence of latex sensitization was 30 %; only two sensitized children (12 %) had symptoms after exposure. Risk factors for latex sensitization were age 5 years (p = 0.008; OR = 6.0; 95% CI = 1.7-22.1), having at least four previous surgical interventions (p < 0.0001; OR = 18.5; 95% CI = 3.6-94.8), having undergone surgery in the first 3 months of life (p = 0.008; OR = 5.4; 95% CI = 0.7-29.2) and total serum IgE 44 IU/ml (p = 0.03; OR = 3.8; 95 %CI = 1.1-13.1). Multiple logistic regression analysis showed that only a history of four or more surgical interventions (p < 0.0001; OR = 26.3; 95 %CI = 2.9-234.2) and total serum IgE 44 IU/ml (p = 0.02; OR = 8.6; 95% CI = 1.4-53.4) were independently associated with latex sensitization. Sex, family and personal allergic history, hydrocephalus with ventriculoperitoneal shunt, cystourethrograms, intermittent bladder catheterization and atopy were not related to latex sensitization. Conclusions: In children with spina bifida, significant and independent risk factors identified for latex sensitization were multiple interventions and higher levels of total serum IgE. A prospective study will clarify the clinical evolution of assymptomatic children sensitized to latex.

Factores de Risco para Náuseas e Vómitos no Pós-Operatório de Cirurgia Oftalmológica

Fundamentação: As náuseas e vómitos no pós-operatório são frequentes em cirurgia oftalmológica e podem ter repercussões graves. Não estão identificados os factores de risco para a sua ocorrência e desconhece-se qual a estratégia profilática mais eficaz. Objectivo: Identificar os factores de risco para náuseas e vómitos no pós-operatório em cirurgia oftalmológica e comparar a eficácia profilática das associações droperidol-metoclopramida e dexametasona-metoclopramida. Metodologia: Estudo prospectivo unicêntrico de 80 doentes sem história prévia de náuseas e vómitos no pós-operatório, propostos para cirurgia oftalmológica sob anestesia geral. Aplicou-se um questionário para caracterização demográfica, clínica e laboratorial. Todos os doentes foram submetidos a profilaxia com droperidol-metoclopramida (1,25+20mg) ou dexametasona-metoclopramida (5+20mg), segundo distribuição aleatorizada por blocos e assegurando dupla-ocultação. Resultados: Ocorreram náuseas e vómitos no pós-operatório em 17,5% dos doentes. O sistema de estratificação de risco de Apfel não identificou os doentes que desenvolveram náuseas e vómitos no pós-operatório. O risco de náuseas e vómitos no pós-operatório foi significativamente maior entre os adultos nos extremos etários (<40 anos e ≥60 anos), naqueles com menor classe de peso e nos doentes com classe ASA-I (OR: 3,36). As náuseas e vómitos no pós-operatório foram menos frequentes entre os doentes medicados com dexametasona-metoclopramida (12,8%) do que naqueles submetidos a profilaxia com droperidol-metoclopramida (22%), mas a diferença não atingiu significado estatístico. Conclusões: Os sistemas de estratificação do risco de náuseas e vómitos no pós-operatório não são aplicáveis em cirurgia oftalmológica, sendo necessários estudos em maior escala neste âmbito. É possível que a associação dexametasona-metoclopramida seja não apenas tão eficaz quanto a droperidol-metoclopramida, como eventualmente superior.

Physical Activity, Obesity Status, and Blood Pressure in Preschool Children

Objective To examine the combined effects of physical activity and weight status on blood pressure (BP) in preschool-aged children. Study design The sample included 733 preschool-aged children (49% female). Physical activity was objectively assessed on 7 consecutive days by accelerometry. Children were categorized as sufficiently active if they met the recommendation of at least 60 minutes daily of moderate-to-vigorous physical activity (MVPA). Body mass index was used to categorize children as nonoverweight or overweight/obese, according to the International Obesity Task Force benchmarks. BP was measured using an automated BP monitor and categorized as elevated or normal using BP percentile-based cut-points for age, sex, and height. Results The prevalence of elevated systolic BP (SBP) and diastolic BP was 7.7% and 3.0%, respectively. The prevalence of overweight/obese was 32%, and about 15% of children did not accomplish the recommended 60 minutes of daily MVPA. After controlling for age and sex, overweight/obese children who did not meet the daily MVPA recommendation were 3 times more likely (OR 3.8; CI 1.6-8.6) to have elevated SBP than nonoverweight children who met the daily MVPA recommendation. Conclusions Overweight or obese preschool-aged children with insufficient levels of MVPA are at significantly greater risk for elevated SBP than their non overweight and sufficiently active counterparts. (J Pediatr 2015;167:98-102).

The relationship of cardiorespiratory fitness, birth weight and parental BMI on adolescents’ obesity status

The aims of this study were as follows: (1) to analyze differences in cardiorespiratory fitness (CRF), parents' body mass index (BMI) and birth weight (BW) between non-overweight (NOW) and overweight/obese (OV/OB) adolescents, and (2) to investigate the association of those variables with the risk of their biological offspring being OV/OB. This study comprised 788 adolescents (477 girls and 311 boys), aged between 12 and 18 years. CRF was predicted by maximal multistage 20-m shuttle-run test according to the procedures described in FITNESSGRAM. Children's BMI was classified according to the International Obesity Task Force. Adolescents' BW was assessed from each child's pediatric record at birth. Parents' OV/OB status was defined and classified according to the World Health Organization. Socioeconomic status was defined by parental education. The prevalence OV/OB was 21.4 and 5.3%, respectively, and there were no gender differences. The OV/OB adolescents (girls and boys) had significantly (P<or=0.05) lower CRF scores and higher BW (P<or=0.05) than did the NOW pears. Overall, 92.9% of OV/OB girls had one or two parents with OV/OB (P<or=0.05). Boys with low CRF (odds ratio (OR): 3.75; P<or=0.05) and high BW (OR: 1.65; P<or=0.05) were more likely to be classified as OV/OB compared with NOW. Girls with low CRF (OR: 2.66; P<or=0.05), high BW (OR: 2.09; P<or=0.05) and at least one parent (OR: 2.28; P<or=0.05) or two parents with OV/OB (OR: 4.39; P<or=0.05, respectively) were classified as OV/OB. Results from this study highlight the association between the family adolescents' obesity, in girls. Furthermore, our data suggested that low CRF and high BW were strong predictors of OV/OB in adolescence.

Factores de Risco para Sensibilização ao Látex em Crianças com Espinha Bífida

OBJECTIVO: Pretendeu-se com este estudo determinar a prevalência e identificar factores de risco para sensibilização ao látex em crianças com espinha bífida. MÉTODOS: Estudaram-se 57 crianças com espinha bífida, com uma idade média de 5.6 anos(6 meses a 18 anos) e uma relação sexo M/F de 0.8/1. A todas as crianças foram efectuados questionário, bateria de TC incluindo látex (extractos UCBStallergenes, Lofarma e ALK-Abelló), aeroalergenos comuns e frutos (UCB-Stallergenes) e determinação sérica de IgE total (AlaSTAT). RESULTADOS: A prevalência de sensibilização ao látex foi de 30%; apenas duas crianças sensibilizadas (12%) apresentavam sintomatologia relacionada com a exposição (urticária/angioedema e rinite). Foram identificados como factores de risco para sensibilização ao látex: idade ≥ 5 anos (p=0.008; OR=6.0, IC95%=1.7-22.1); existência de 4 ou mais intervenções cirúrgicas (p<0.0001; OR=18.5, IC95%=3.6-94.8); cirurgias nos primeiros 3 meses de vida (p=0.008; OR=5.4, IC95%=0.7-29.2); níveis séricos de IgE total ≥ 44UI/ml (p=0.03; OR=3.8, IC95%=1.1-13.1). Pela realização de regressão logística foram identificados como factores de risco independentes, história de 4 ou mais cirurgias (p<0.0001; OR=26.3, IC95%=2.9-234.2) e níveis de IgE total ≥ 44UI/ml (p=0.02; OR=8.6, IC95%=1.4-53.4). Sexo, antecedentes familiares e pessoais de patologia alérgica, hidrocefalia com derivação ventriculo-peritoneal, cistografias,cateterismo vesical intermitente e atopia não foram identificados como factores de risco. CONCLUSÕES: Identificámos como factores de risco significativos e independentes para sensibilização ao látex em crianças com espinha bífida a existência de número elevado de intervenções cirúrgicas (≥ 4 cirurgias) e níveis séricos mais elevados de IgE total(IgE total ≥ 44UI/ml). Estudo prospectivo esclarecerá a evolução clínica das crianças sensibilizadas assintomáticas.

HISTOPLASMIN SURVEY IN HIV-POSITIVE PATIENTS: RESULTS FROM AN ENDEMIC AREA IN NORTHEASTERN BRAZIL

SUMMARY Background. Disseminated histoplasmosis is common in AIDS patients with advanced immunosuppression in Ceará, Northeastern Brazil. The goal of this study was to determine the prevalence of Histoplasma infection in patients with HIV/AIDS living in Fortaleza, the capital of Ceará. Methods. Intradermal tests with histoplasmin (mycelial phase) were performed in 161 HIV patients with CD4 ≥ 350 cells/mm 3 . Evidence of recent illness was evaluated with immunodiffusion (ID) tests in 76 of these individuals. Results. A total of 11.8% of patients reacted to histoplasmin and 2.63% had ID test positive to Histoplasma. The presence of mango trees (Mangifera indica) in the patient neighborhood (OR = 2.870; 95% CI = 1.081-7.617; p = 0.040) and past activity involving soil (OR = 2.834; 95% CI = 1.045-7.687; p = 0.045) or visits to a farm (OR = 3.869; 95% CI = 1.189-12.591; p = 0.033) were significantly associated with Histoplasma infection. Conclusions. Patients with HIV living in Fortaleza have an expressive prevalence of infection with Histoplasma.

Factores de Risco para Asma Activa em Idade Escolar: Estudo Prospectivo com Oito Anos de Duração

A sibilância recorrente na infância é uma entidade clínica prevalente e heterogénea do pontode vista da história natural e do prognóstico. Efectuou-se um estudo prospectivo com 8 anos de duração, com o objectivo de relacionar a evolução clínica da sibilância recorrente nos primeiros anos de vida, com factores de prognóstico associados com a persistência da sintomatologia. Uma coorte de 308 crianças com sibilância recorrente, com idade ≤6 anos, foi incluída no estudo em 1993. Foi aplicado um questionário clínico, realizados testes cutâneos por prick e efectuado doseamento sérico de IgE total. Em 1996 procedeu-se a uma primeira reavaliação sistemática destas crianças. Em 2001 foi efectuada nova reavaliação sistemática, possível em 81% destas crianças (n=249), com repetição dos testes cutâneos e realização de avaliação funcional respiratória,em período intercrise, com espirometria com prova de broncodilatação (BD). As crianças reavaliadas apresentavam média etária de 11 anos (8-14 anos) e relação sexo M/F de 1.7/1. Permaneciam sintomáticas em 61% dos casos. A prevalência de atopia foi de 48% em 1993, 65% em 1996 e 75% em 2001. Pela realização de um modelo de regressão logística múltiplo foram identificados como factores de risco para asma activa em idade escolar: história pessoal de rinite alérgica (OR=15.8, IC95%=6.1-40.8; p<0.001), asma paterna (OR=7.2, IC95%=1.7-29.7; p=0.007), história pessoal de eczema atópico (OR=5.9, IC95%=2.2-15.7; p<0.001), asma materna (OR=5.4, IC95%=1.7-17.1; p=0.004), evidência de sensibilização alergénica (OR=3.4, IC95%=1.2-10.4;p=0.03) e início dos sintomas ≥2 anos de idade (OR=2.1, IC95%=1.1-4.8; p=0.04); a frequência de infantário antes dos 12 meses de idade foi identificada como factor protector (OR=0.4, IC95%=0.2- 0.9; p=0.04). Desenvolveram sensibilização alergénica de novo (ácaros do pó >80%) 66 das 128 crianças não atópicas em 1993 (52%). Apresentavam obstrução brônquica 36% das crianças: 47% das sintomáticas e 18% das assintomáticas (p<0.001). A prova de BD foi positiva em 35%: 47% nos sintomáticos e 13% nos assintomáticos (p<0.001). Concluindo, foram identificados como factores de mau prognóstico, antecedentes pessoais de doença alérgica, história parental de asma, presença de sensibilização alergénica e início dos sintomas na segunda infância. Os sintomas clínicos podem preceder em anos a sensibilização alergénica, realçando a importância da instituição precoce de medidas de controlo ambiental. Alterações nas provas funcionais respiratórias, mais frequentes nas asmas activas, estavam também presentes em crianças actualmente sem clínica, reforçando a necessidade de valorizar marcadores objectivos nesta cada vez mais prevalente doença respiratória crónica.

HAND-FOOT-AND-MOUTH DISEASE EPIDEMIOLOGICAL STATUS AND RELATIONSHIP WITH METEOROLOGICAL VARIABLES IN GUANGZHOU, SOUTHERN CHINA, 2008-2012

Hand-foot-and-mouth disease (HFMD) is becoming one of the extremely common airborne and contact transmission diseases in Guangzhou, southern China, leading public health authorities to be concerned about its increased incidence. In this study, it was used an ecological study plus the negative binomial regression to identify the epidemic status of HFMD and its relationship with meteorological variables. During 2008-2012, a total of 173,524 HFMD confirmed cases were reported, 12 cases of death, yielding a fatality rate of 0.69 per 10,000. The annual incidence rates from 2008 to 2012 were 60.56, 132.44, 311.40, 402.76, and 468.59 (per 100,000), respectively, showing a rapid increasing trend. Each 1 °C rise in temperature corresponded to an increase of 9.47% (95% CI 9.36% to 9.58%) in the weekly number of HFMD cases, while a one hPa rise in atmospheric pressure corresponded to a decrease in the number of cases by 7.53% (95% CI -7.60% to -7.45%). Similarly, each one percent rise in relative humidity corresponded to an increase of 1.48% or 3.3%, and a one meter per hour rise in wind speed corresponded to an increase of 2.18% or 4.57%, in the weekly number of HFMD cases, depending on the variables considered in the model. These findings revealed that epidemic status of HFMD in Guangzhou is characterized by high morbidity but low fatality. Weather factors had a significant influence on the incidence of HFMD.

CHICKEN COOPS, Triatoma dimidiata INFESTATION AND ITS INFECTION WITH Trypanosoma cruzi IN A RURAL VILLAGE OF YUCATAN, MEXICO

This study longitudinally investigated the association between Triatoma dimidiata infestation, triatomine infection with Trypanosoma cruzi and household/backyard environmental characteristics in 101 homesteads in Molas and Yucatan, Mexico, between November 2009 (rainy season) and May 2010 (dry season). Logistic regression models tested the associations between insect infestation/infection and potential household-level risk factors. A total of 200 T. dimidiata were collected from 35.6% of the homesteads, mostly (73%) from the peridomicile. Of all the insects collected, 48% were infected with T. cruzi. Infected insects were collected in 31.6% of the homesteads (54.1% and 45.9% intra- and peridomiciliary, respectively). Approximately 30% of all triatomines collected were found in chicken coops. The presence of a chicken coop in the backyard of a homestead was significantly associated with both the odds of finding T. dimidiata (OR = 4.10, CI 95% = 1.61-10.43, p = 0.003) and the presence of triatomines infected with T. cruzi (OR = 3.37, CI 95% = 1.36-8.33, p = 0.006). The results of this study emphasize the relevance of chicken coops as a putative source of T. dimidiata populations and a potential risk for T. cruzi transmission.