Fluconazole and amphotericin-B resistance are associated with increased catalase and superoxide dismutase activity in Candida albicans and Candida dubliniensis

Introduction Candida dubliniensis, a new species of Candida that has been recovered from several sites in healthy people, has been associated with recurrent episodes of oral candidiasis in AIDS and HIV-positive patients. This species is closely related to C. albicans. The enzymatic activity of C. dubliniensis in response to oxidative stress is of interest for the development of drugs to combat C. dubliniensis. Methods Fluconazole- and amphotericin B-resistant strains were generated as described by Fekete-Forgács et al. (2000). Superoxide dismutase (SOD) and catalase assays were performed as described by McCord and Fridovich (1969) and Aebi (1984), respectively. Results We demonstrated that superoxide dismutase (SOD) and catalase activities were significantly higher (p<0.05) in the fluconazole- and amphotericin B-resistant strains of C. dubliniensis and C. albicans than in the sensitive strains. The catalase and SOD activities were also significantly (p<0.01) higher in the sensitive and resistant C. albicans strains than in the respective C. dubliniensis strains. Conclusions These data suggest that C. albicans is better protected from oxidative stress than C. dubliniensis and that fluconazole, like amphotericin B, can induce oxidative stress in Candida; oxidative stress induces an adaptive response that results in a coordinated increase in catalase and SOD activities.

Epidemiology of hepatitis B virus infection among recyclable waste collectors in central Brazil

Introduction: The collection of recyclable waste materials is a widespread activity among the urban poor. Today, this occupation attracts an increasingly large number of individuals. Despite its economic and environmental importance, this activity is associated with unsafe and unhealthy working conditions. The aim of this study was to investigate the seroepidemiological profile of hepatitis B virus (HBV) infection in a population of recyclable waste collectors in central Brazil. Methods: Recyclable waste collectors from all 15 recycling cooperatives in Goiânia City were invited to participate in the study. The participants (n = 431) were interviewed and screened for hepatitis B surface antigen (HBsAg) and antibodies against HBsAg (anti-HBs) and hepatitis B core antigen (anti-HBc) by enzyme-linked immunosorbent assay (ELISA). HBsAg- and anti-HBc-positive samples were tested for HBV DNA and genotyped. Results: The overall prevalence of HBV infection (HBsAg- and/or anti-HBc-positive) was 12.8%. An age over 40 years and illicit drug use were associated with HBV infection. HBV DNA was detected in 2/3 HBsAg-positive samples and in 1/52 anti-HBc-positive/HBsAg-negative samples (an occult HBV infection rate of 1.9%), in which the genotypes/subgenotypes A/A1, D/D3 and F/F2 were identified. Only 12.3% of the recyclable waste collectors had serological evidence of previous HBV vaccination. Conclusions: These findings highlight the vulnerability of recyclable waste collectors to HBV infection and reinforce the importance of public health policies that address the health and safety of this socially vulnerable population.

Hepatitis B virus genotypes and mutations in the basal core promoter and pre-core/core in chronically infected patients in southern Brazil: a cross-sectional study of HBV genotypes and mutations in chronic carriers

Introduction In Brazil, little data exist regarding the distribution of genotypes in relation to basal core promoter (BCP) and precore/core mutations among chronic hepatitis B virus (HBV) carriers from different regions of the country. The aim of this study was to identify HBV genotypes and the frequency of mutations at the BCP and precore/core region among the prevalent genotypes in chronic carriers from southern Brazil. Methods Nested-polymerase chain reaction (nested-PCR) products amplified from the S-polymerase gene, BCP and precore/core region from 54 samples were sequenced and analyzed. Results Phylogenetic analysis of the S-polymerase gene sequences showed that 66.7% (36/54) of the patients were infected with genotype D (D1, D2, D3), 25.9% (14/54) with genotype A (A1, A2), 5.6% (3/54) with subgenotype C2, and 2% (1/54) with genotype E. A comparison of virological characteristics showed significant differences between genotypes A, C and D. The comparison between HBeAg status and the G1896A stop codon mutation in patients with genotype D revealed a relationship between HBV G1896A precore mutants and genotype D and hepatitis B e antigen (HBeAg) seroconversion. Genotype D had a higher prevalence of the G1896A mutation and the presence of a thymine at position 1858. Genotype A was associated with a higher prevalence of the G1862T mutation and the presence of a cytosine at position 1858. Conclusions HBV genotype D (D3) is predominant in HBV chronic carriers from southern Brazil. The presence of mutations in the BCP and precore/core region was correlated with the HBV genotype and HBeAg negative status.

Epidemiological study of hepatitis B and C in a municipality with rural characteristics: Cássia dos Coqueiros, State of São Paulo, Brazil

Abstract: INTRODUCTION: Hepatitis B and C viral infections remain an important cause of global morbidity and mortality. Studies have been conducted in population groups of large cities, leaving gaps in the knowledge regarding the situation in small municipalities. We aimed to measure the prevalence of hepatitis B and C markers and presence of infection-associated factors. METHODS: All inhabitants of Cássia dos Coqueiros aged ≥18 years who agreed to participate in the research were included. We collected blood as well as information via a questionnaire between March 2011 and December 2013. Univariate and multivariate analyses were conducted. RESULTS: Among the 1,001 participants, 41 (4.1%) participants had a serological profile of hepatitis B viral exposure, and only one (0.1%) participant was considered a virus carrier. The frequency of isolated antibody to hepatitis B virus surface antigen (anti-HBs) markers was 17.8% for the overall population. In the multivariate analysis, hepatitis B virus (HBV) infection was associated with age, birth outside the State of São Paulo, history of hepatitis, ≥2 sexual partners in the last 6 months, and tattoos. Four (0.4%) participants had a serological profile of hepatitis C viral exposure. However, after confirmation using viral ribonucleic acid (RNA) evaluation, only one (0.1%) individual remained positive. CONCLUSIONS: The positivity rates for hepatitis B and C were low, despite greater sexual freedom and the recent emergence of illicit drugs, as observed by the health personnel working in Cássia dos Coqueiros.

Clinical, demographic, and epidemiologic characteristics of hepatitis B virus-infected patients at a tertiary public hospital in Presidente Prudente, State of São Paulo, Brazil

Abstract: INTRODUCTION: Few studies have addressed the primary characteristics of patients infected with hepatitis B virus (HBV) in the general population, especially those living in small- and medium-sized cities in Brazil. We aimed to determine the clinical, demographic, and epidemiologic characteristics of patients diagnosed with HBV who were followed up at an infectious diseases clinic of a public hospital in State of São Paulo, Brazil. METHODS: Medical records of patients aged >18 years and diagnosed with HBV infection between January 2000 and December 2013 were reviewed. RESULTS: Seventy-five patients were enrolled with male-female main infection-associated risk factors; 9 (12%) were co-infected with human immunodeficiency virus (HIV), 5 (6.7%) with hepatitis C virus (HCV), and 3 (4%) were co-infected with both HIV and HCV. Antiviral HBV therapy was applied in 21 (28%) patients and tenofovir monotherapy was the most prescribed medication. After approximately 2 years of antiviral treatment, the HBV-DNA viral load was undetectable in 12 (92.3%) patients and lower levels of alanine aminotransferase were found in these patients. CONCLUSIONS: Over a 13-year interval, very few individuals infected with HBV were identified, highlighting the barriers for caring for patients with HBV in developing countries. New measures need to be implemented to complement curative practices.

Friedreich's ataxia: clinical and molecular study of 25 Brazilian cases

INTRODUCTION: Friedreich's ataxia is a neurodegenerative disorder whose clinical diagnostic criteria for typical cases basically include: a) early age of onset (< 20 or 25 years), b) autosomal recessive inheritance, c) progressive ataxia of limbs and gait, and d) absence of lower limb tendon reflexes. METHODS: We studied the frequency and the size of expanded GAA and their influence on neurologic findings, age at onset, and disease progression in 25 Brazilian patients with clinical diagnosis of Friedreich's ataxia - 19 typical and 6 atypical - using a long-range PCR test. RESULTS: Abnormalities in cerebellar signs, in electrocardiography, and pes cavus occurred more frequently in typical cases; however, plantar response and speech were more frequently normal in this group when the both typical and atypical cases were compared. Homozygous GAA expansion repeats were detected in 17 cases (68%) - all typical cases. In 8 patients (32%) (6 atypical and 2 typical), no expansion was observed, ruling out the diagnosis of Friedreich's ataxia. In cases with GAA expansions, foot deformity, cardiac abnormalities, and some neurologic findings occurred more frequently; however, abnormalities in cranial nerves and in tomographic findings were detected less frequently than in patients without GAA expansions. DISCUSSION: Molecular analysis was imperative for the diagnosis of Friedreich's ataxia, not only for typical cases but also for atypical ones. There was no genotype-phenotype correlation. Diagnosis based only on clinical findings is limited; however, it aids in better screening for suspected cases that should be tested. Evaluation for vitamin E deficiency is recommended, especially in cases without GAA expansion.

Use of nonradioactive labeling to detect large gene rearrangements in 21-hydroxylase deficiency

PURPOSE: To establish the Southern blotting technique using hybridization with a nonradioactive probe to detect large rearrangements of CYP21A2 in a Brazilian cohort with congenital adrenal hyperplasia due to 21-hydroxylase deficiency (CAH-21OH). METHOD: We studied 42 patients, 2 of them related, comprising 80 non-related alleles. DNA samples were obtained from peripheral blood, digested by restriction enzyme Taq I, submitted to Southern blotting and hybridized with biotin-labeled probes. RESULTS: This method was shown to be reliable with results similar to the radioactive-labeling method. We found CYP21A2 deletion (2.5%), large gene conversion (8.8%), CYP21AP deletion (3.8%), and CYP21A1P duplication (6.3%). These frequencies were similar to those found in our previous study in which a large number of cases were studied. Good hybridization patterns were achieved with a smaller amount of DNA (5 mug), and fragment signs were observed after 5 minutes to 1 hour of exposure. CONCLUSIONS: We established a non-radioactive (biotin) Southern blot/hybridization methodology for CYP21A2 large rearrangements with good results. Despite being more arduous, this technique is faster, requires a smaller amount of DNA, and most importantly, avoids problems with the use of radioactivity.

Observações sobre autocompatibilidade em pupunha (Bactris gasipaes H.B.K., Palmae)

A pupunha aparentemente, possui duas estratégias que asseguram a sua polinização cruzada. Uma delas parece ser um fator genético quantitativo que inibe a auto-polinização. Para estudar este fator, duas series de polinização controlada foram feitas em duas diferentes populações juvenis. A primeira população apresentou uma media, de 19,2% de auto-compatibilidade, medida em percentagem de frutos férteis produzidos em relação a todas as flores. Entretanto, o coeficiente de variação foi extremamente alto. A polinização aberta em algumas plantas produziu 22% de frutos férteis, com um coeficiente de variação menor, sugerindo que, o " fruit set" foi, em geral, baixo nutria população, naquela vez. A segunda população, apresentou em media 37,62% de auto-compatibilidade, com um coeficiente de variação menor. Variação entre as árvores foi também detectada e mostrou ser considerável. Além do mais, a auto-compatibilidade variou de 0 a 81%, em distribuição normal. 0 uso de plantas juvenis pode ser a razão da alta variabilidade entre plantas. Apesar da pouca auto-compatibilidade encontrada nestas populações, esta característica é recomendada como descritor para bancos de germoplasma e deve ser obtida em plantas adultas para evitar a alta variabilidade juvenil.

Açaí (Euterpe oleracea mart.) e camu-camu (Myrciaria dubia (H.B.K.) Mc Vaugh) possuem ação anti anêmica?

Avaliou-se o impacto da utilização do açaí e camu-camu em pré-escolares de uma Unidade Filantrópica de Manaus-AM. Foram selecionadas 85 crianças voluntárias, de dois a seis anos incompletos, de ambos os sexos, distribuídas aleatoriamente em 5 grupos, tendo como fonte de ferro e vitamina C: açaí e açaí + camu-camu, perfazendo um total de 2 mg de ferro e 40 mg de ácido ascórbico, assim como o ferro aminoácido quelato na concentração de 1 e 2 mg de ferro. O ferro foi distribuído diariamente na colação por um período de 120 dias. Na caracterização da anemia considerou-se o ponto de corte de hemoglobina <11g/dL. Os resultados demonstraram que de um universo de 85 crianças 6 (7%) apresentaram um quadro de desnutrição crônica, sendo ao final da intervenção reduzido para 4 (4,7%). O maior impacto da utilização do açaí foi como fonte energética, refletido no ganho de peso significativo das crianças (1,76 kg), mesmo quando adicionado de camu-camu (1,69 kg). Em relação a concentração de hemoglobina, não foi constatada diferença significativa entre as crianças dos diferentes grupos, independente da fonte de ferro: açaí (0,71 g/dL); açaí + camu-camu (0,60 g/dL), Ferro 2 mg (0,88 g/dL); água (0,85 g/ dL) e Ferro 1 mg (0,54 g/dL). Entretanto, a recuperação de crianças anêmicas foi maior no grupo que recebeu ferro aminoácido quelato na concentração de 2 mg de ferro. Conclui-se que o açaí tem um grande potencial como fonte energética e pouca expressividade como fonte de ferro, mesmo adicionado de camu-camu.

Physiological responses of matrinxã (Brycon amazonicus) fed different levels of vitamin C and submitted to air exposure

The role of vitamin C on physiological responses of matrinxã (Brycon amazonicus) submitted to air exposure was analyzed. Nine hundred fish (70.15 g) were distributed in fifteen 500 l boxes (60 fish.box-1) and fed five rations (treatments): Control (no vitamin C); T100 (100 mg); T200 (200 mg); T400 (400 mg) and T800 (800 mg of vitamin C kg.ration-1). Each ration was offered to fish of three boxes during 60 days before the stress challenge that consisted of exposing fish to air for two minutes. Samplings were carried out for 5, 15, 30 and 60 minutes after the air exposure. Blood was collected for glucose, cortisol, total protein, sodium, chloride, hematocrit, hemoglobin determination, and white and red cell count. Liver was removed for hepatosomatic index (HSI) calculation and glycogen determination. Vitamin C did not affect the levels of cortisol, chloride, total protein, hemoglobin, leukocytes, hepatic glycogen or HSI in air exposed fish. Blood glucose levels elevation observed 60 minutes after the challenge did not depend on the levels of vitamin C, nor did the drop in serum sodium levels verified 60 minutes after stressor. In general, hematocrit did not change by effect of vitamin C but it was lower at 15 and 30 minutes after the challenge. The number of erythrocytes decreased in fish after 5 minute sampling in all treatments, especially at 30 and 60 minutes. The air exposure evoked alterations in stress indicators of matrinxã, and the vitamin C did not alter the responses.

Constituintes químicos fixos e voláteis dos talos e frutos de Piper tuberculatum Jacq. e das raízes de P. hispidum H. B. K.

Os óleos essenciais dos frutos e talos finos de Piper tuberculatum e das raízes de P. hispidum, coletados no estado de Rondônia, foram obtidos por hidrodestilação e analisados por GC e GC-MS. Foram identificados como constituintes majoritários, nos óleos dos frutos e talos finos de P. tuberculatum, o óxido de cariofileno (32,1%) e (26,6%) e o (E)-cariofileno (17,7%) e (12,3%), respectivamente. No óleo essencial das raízes de P. hispidum, foram identificados, como constituintes majoritários, o dilapiol (57,5%), a elemicina (24,5%) e o apiol (10,2%). Do extrato etanólico dos frutos de P. tuberculatum, foram isolados os esteróides β-sitosterol e estigmasterol, as amidas piplartina e dihidropiplartina e um derivado do ácido cinâmico, o ácido 3,4,5-trimetoxi-dihidrocinâmico.

Propagação vegetativa por miniestacas de preciosa (Aniba canellila (H. B.K) MEZ)

Este trabalho teve como objetivo o enraizamento de miniestacas de material juvenil da preciosa (Aniba canelilla (H.B.K) Mez), tratadas com diferentes concentrações de ácido indol-3-butirico (AIB) na forma líquida. As miniestacas foram obtidas de mudas de regeneração natural com aproximadamente um ano de idade e modeladas com 5 cm de comprimento e 0,4 - 0,6 mm de diâmetro. O experimento foi conduzido em casa de vegetação, com sistema de nebulização intermitente regulada em 20 segundos para aspersões com intervalos de 20 minutos. Como substrato foi utilizado areia lavada. Semanalmente foram feitas aplicações de fertilizante foliar e fungicida. O experimento foi delineado em cinco blocos inteiramente ao acaso, com cinco tratamentos (0, 300 ppm, 600 ppm, 1200 ppm e 2000 ppm de AIB) com 25 miniestacas/tratamento, totalizando 125 miniestacas no experimento. Após 180 dias do plantio, as miniestacas foram retiradas do substrato e avaliados os seguintes parâmetros: porcentagem de enraizamento, porcentagem de sobrevivência, porcentagem de calo; porcentagem de brotos; número de raízes principais, comprimento médio das raízes; peso da matéria seca dos brotos e peso da matéria seca das raízes. Os resultados indicam que a emissão de raízes das estacas de material juvenil da preciosa independe do uso do AIB. Entretanto, o uso desta auxina na concentração de 2000 ppm estimulou o enraizamento (79,04 %); sobrevivência (89,43%) e brotação (64%) das miniestacas.

The Role of B-Type Natriuretic Peptide in the Diagnosis of Congestive Heart Failure in Patients Presenting to an Emergency Department with Dyspnea

OBJECTIVE: To determine the utility of B-type natriuretic peptide (BNP) in the diagnosis of congestive heart failure (CHF) in patients presenting with dyspnea to an emergency department (ED). METHODS: Seventy patients presenting with dyspnea to an ED from April to July 2001 were included in the study. Mean age was 72±16 years and 33 (47%) were male. BNP was measured in all patients at the moment of admission to the ED. Emergency-care physicians, blinded to BNP values, were required to assign a probable initial diagnosis. A cardiologist retrospectively reviewed the data (blinded to BNP measurements) and assigned a definite diagnosis, which was considered the gold standard for assessing the diagnostic performance of BNP. RESULTS: The mean BNP concentration was higher in patients with CHF (n=36) than in those with other diagnoses (990±550 vs 80±67 pg/mL, p<0.0001). Patients with systolic dysfunction had higher BNP levels than those with preserved systolic function (1,180±641 vs 753±437 pg/mL, p=0.03). At a blood concentration of 200 pg/mL, BNP showed a sensitivity of 100%, specificity of 97.1%, positive predictive value of 97.3%, and negative predictive value of 100%. The application of BNP could have potentially corrected all 16 cases in which the diagnosis was missed by the emergency department physician. CONCLUSION: BNP measurement is a useful tool in the diagnosis of CHF in patients presenting to the ED with dyspnea.

Sensibilidade e especificidade do hormônio natriurético do tipo B para identificar doentes com insuficiência mitral grave sintomáticos e assintomáticos

OBJETIVO: Verificar a sensibilidade e a especificidade do hormônio natriurético do tipo B (BNP) para identificar pacientes ambulatoriais, com insuficiência mitral crônica grave, sintomáticos e assintomáticos. MÉTODOS: Um grupo de pacientes com insuficiência mitral foi examinado e submetido à eletrocardiografia, telerradiografia de tórax, coletas de sangue venoso e ecocardiograma transtorácico. Por meio da análise de variáveis ecocardiográficas, 62 pacientes apresentavam refluxo mitral discreto e moderado (G I) e 34 refluxo mitral grave (G II). A capacidade discriminante do BNP em detectar pacientes com insuficiência mitral grave foi avaliada pela construção de curvas ROC. RESULTADOS: Entre os 96 doentes, 71 (73%) eram mulheres e as idades variaram entre 15 e 63 (média de 31,7) anos. Os valores de BNP variaram de 0,00 pg/ml a 193 pg/ml. Os doentes do G I tiveram um valor médio de BNP de 18,10 ± 0,74 pg/ml e os do G II de 50,54 ± 1,46 pg/ml, (p=0,001). O valor de corte para identificar insuficiência mitral grave foi de 15,40 pg/ml, para o melhor balanço entre a sensibilidade e a especificidade, respectivamente de 0,73 e 0,74. O valor de corte para identificar pacientes sintomáticos e com insuficiência mitral grave foi de 28,40 pg/ml, para o melhor balanço entre a sensibilidade e a especificidade, respectivamente de 0,78 e 0,83. CONCLUSÃO: Os valores de BNP capazes de indentificar doentes com insuficiência mitral grave assintomáticos e sintomáticos são menores do que os 100 pg/ml considerados para o diagnóstico de insuficiência cardíaca.

Avaliação de pacientes assintomáticos com forma crônica da doença de Chagas através da análise do eletrocardiograma dinâmico, ecocardiograma e do peptídeo natriurético tipo B

OBJETIVO: Avaliar pacientes assintomáticos com forma crônica da doença de Chagas em relação a prevalência de arritmias ventriculares, disfunção ventricular esquerda e níveis plasmáticos do peptídeo natriurético tipo B (BNP). MÉTODOS: Avaliação clínica, eletrocardiograma (ECG), índice cardiotorácico (ICT), eletrocardiograma dinâmico, ecocardiograma e dosagem BNP foram realizados em 106 pacientes do Ambulatório de Doença de Chagas, distribuídos em três grupos: GI (50-ECG normal), GIIA (31-ECG com alterações características de doença de Chagas) e GIIB (25-ECG com outras alterações). RESULTADOS: Alterações eletrocardiográficas mais prevalentes no GIIA: bloqueio completo do ramo direito, bloqueio divisional ântero-superior esquerdo (35% cada) e áreas inativas (32%), GIIB: alteração da repolarização inferolateral (28%) e sobrecarga ventricular esquerda (24%). Os valores médios do ICT foram semelhantes (p = 0,383). A prevalência de arritmia ventricular foi maior nos grupos GIIA (77%) e GIIB (75%) do que no GI (46%) (p = 0,002). A disfunção ventricular foi mais prevalente no GIIA (52%) e GIIB (32%) do que no GI (14%) (p = 0,001). A disfunção sistólica foi mais prevalente no GIIA (29%) do que no GIIB (20%) e GI (2%) (p < 0,001). A disfunção diastólica foi mais prevalente no GIIA (42%) e no GIIB (28%) do que no GI (12%) (p = 0,005). Os valores médios do peptídeo natriurético tipo B foram, respectivamente, 30 ± 88 pg/ml no GI, 66 ± 194 no GIIA e 24 ± 82 no GIIB (p = 0,121). CONCLUSÃO: Pacientes assintomáticos com forma crônica da doença de Chagas e ECG alterado têm maior prevalência de arritmias e disfunção ventricular esquerda do que pacientes com ECG normal. Os níveis plasmáticos do BNP foram semelhantes entre os grupos.