Hibridização reversa e sequenciamento na genotipagem do vírus da hepatite C

INTRODUÇÃO: Os métodos de genotipagem do vírus da hepatite C têm sido muito discutidos. O objetivo deste trabalho foi comparar as metodologias de hibridização reversa e sequenciamento direto para a genotipagem do vírus da hepatite C. MÉTODOS: Noventa e uma amostras de plasma de pacientes assistidos na Faculdade de Medicina de Botucatu da Universidade Estadual Paulista foram utilizadas. A genotipagem por hibridização reversa foi realizada utilizando o kit comercial INNO-LiPA® v.1.0. O sequenciamento direto foi efetuado em sequenciador automático utilizando protocolos in house. RESULTADOS: A genotipagem por sequenciamento direto mostrou-se eficiente na resolução dos resultados inconclusivos pelo kit comercial. O kit mostrou resultados errôneos em relação à subtipagem viral. Além disso, a genotipagem por sequenciamento direto revelou um erro do kit com relação à determinação genotípica questionando a eficiência do método também para a identificação do genótipo viral. CONCLUSÕES: A genotipagem realizada por meio de sequenciamento direto permite uma maior acurácia na classificação viral quando comparada à hibridização reversa.

Level of behavior and knowledge concerning human papillomavirus among university students of a nursing college

INTRODUCTION: Human pappilomavirus is one of the most common sexually transmitted diseases, and persistent HPV infection is considered the most important cause of cervical cancer. It is detected in more than 98% of this type of cancer. This study aimed to determine the level of knowledge concerning human papillomavirus among nursing college students of a private educational institution located in the City of Bauru, SP, and correlate their knowledge according to the course year. METHODS: A descriptive study with a quantitative approach, performed with a questionnaire that permitted the quantification of data and opinions, thus guaranteeing the precision of the results without distortions in analysis or interpretation. The survey was applied to randomly selected 1st, 2nd, 3rd, and 4th-year nursing college students. Twenty students from each level were selected during August 2009, totaling 80 students of both genders. RESULTS: Observation revealed that 4th-year students had greater knowledge than 1st-year students, reflecting the greater period of study, the lack of knowledge of 1st-year students was due to the low level of information acquired before entering college. CONCLUSIONS: The need for complementary studies which determine the profile and knowledge of a larger number of teenagers in relation to HPV was established. The need for educational programs that can overcome this lack of information is undeniable, especially those aimed at making adolescents less susceptible to HPV and other STDs.

Hanseníase no Município de Buriticupu, Estado do Maranhão: busca ativa de casos na população adulta

INTRODUÇÃO: O estudo foi desenvolvido com o objetivo de avaliar a situação da hanseníase na população adulta do Município de Buriticupu, MA. MÉTODOS: Foi empregado o método de busca ativa na identificação de casos novos, no período de 2005 a 2007. Baciloscopia de raspado intradérmico foi feita em todos os pacientes com lesões de pele compatíveis com hanseníase, enquanto exame histopatológico foi feito naqueles em que havia dúvidas na definição da forma clínica. RESULTADOS: Participaram do estudo 15.409 indivíduos, tendo o exame clínico definido o diagnóstico de hanseníase em 62 pacientes, o que representa um coeficiente de detecção de 40,23/10.000. A baciloscopia foi positiva em seis pacientes, sendo predominante a forma clínica tuberculóide, em 31 casos, seguida da forma indeterminada em 20, dimorfa em 10 e virchowiana em um. O estudo permitiu, ainda, a identificação de outras doenças da pele, tais como pitiríase versicolor, dermatofitose, escabiose, vitiligo e carcinoma de pele. CONCLUSÕES: O elevado coeficiente de detecção define o município como hiperendêmico para a hanseníase. A busca ativa de casos é um método importante para o controle da doença.

Neurological involvement in visceral leishmaniasis: case report

Visceral leishmaniasis is a severe and potentially fatal vector-borne disease. The most typical symptoms are fever, hepatosplenomegaly, weight loss, bleeding and bacterial infections. Neurological changes are rarely reported. This paper describes a child who presented with neurological signs as the first symptoms of leishmaniasis; tone was diminished and tremors in the extremities were observed. A diagnosis of visceral leishmaniasis was confirmed by parasite detection in the bone marrow. Symptoms were reversed by specific treatment. The nature of a possible mechanism of neurological involvement in visceral leishmaniasis remains unexplained.

Cryptococcus gattii molecular type VGII as agent of meningitis in a healthy child in Rio de Janeiro, Brazil: report of an autochthonous case

Cryptococcus gattii causes meningoencephalitis in immunocompetent hosts, occurring endemically in some tropical and subtropical regions. Recently, this fungus was involved in an outbreak in Vancouver Island and British Columbia (Canada). In this temperate region, the VGII type is predominant. The paper describes an autochthonous case of meningoencephalitis by C. gattii VGII in a previously health child in Rio de Janeiro, considered nonendemic region of Brazil. The fungus was identified by biochemical tests and the molecular type was determined by URA5-RFLP. The present report highlights the need for clinical vigilance for primary cryptococcal meningitis in nonendemic areas.

Molecular characterization of rabies virus isolated from non-haematophagous bats in Brazil

INTRODUCTION: Rabies is an important zoonosis that causes thousands of deaths worldwide each year. Although the terrestrial cycle, mainly transmitted by dogs, is controlled in Brazil, the aerial cycle remains a serious public health issue, besides the economic problem. In the aerial cycle, the haematophagous bat Desmodus rotundus is the main source of infection, where several different species of non-haematophagous bats can be infected and can transmit the virus. METHODS: The aim of this work was to study the epidemiological pattern of rabies using antigenic characterization with monoclonal antibodies and genetic characterization by reverse-transcriptase polymerase chain reaction followed by sequencing and phylogenetic analysis of non-haematophagous bats' and herbivorous animals' central nervous system samples from the western region of the State of São Paulo, Brazil. RESULTS: From 27 samples, 3 antigenic variants were identified: AgV-3, AgV-4, and AgV-6; and from 29 samples, 5 different clusters were identified, all belonging to the rabies virus species. CONCLUSIONS: Although only non-haematophagous bats were evaluated in the studied region, the majority of samples were from antigenic and genetic variants related to haematophagous bats Desmodus rotundus. Samples from the same antigenic variant were segregated in more than one genetic cluster. This study demonstrated the diversity of rabies virus genetic lineages presented and circulating in non-haematophagous bats in the studied region.

Incidence of congenital toxoplasmosis in the city of Belém, state of Pará, northern Brazil, determined by a neonatal screening program: preliminary results

INTRODUCTION: The aim of this study was to determinate the incidence of congenital toxoplasmosis among a group of newborns (NBs) from Belém using neonatal screening. METHODS: Among the 6,000 newborns referred for investigation of genetic and metabolic diseases, 1,000 were selected for screening for congenital toxoplasmosis by determining the amount of IgM in the eluates of blood collected on filter paper. Positive tests were confirmed using paired serology of the NB and his mother. RESULTS: Out of the 1,000 NBs assessed, one had a positive screening result that was confirmed by paired serology. CONCLUSIONS: The incidence of congenital toxoplasmosis in Belém was 10/10,000 live NBs.

High prevalence of HTLV-1 and 2 viruses in pregnant women in São Luis, state of Maranhão, Brazil

INTRODUCTION: Human T cell lymphotropic virus type 1 (HTLV-1) is endemic in the Caribbean, Japan, South America and regions of Africa. HTLV-2 is present in Native American populations and associated with IV drug use in Europe and North America. In Brazil, it is estimated that 1.5 million people are infected with HTLV-1/2. The study objective was to determine HTLV-1/2 prevalence in pregnant women in the prenatal care from three public services in São Luis, State of Maranhão, Brazil, and to counsel seropositive women to reduce viral transmission. METHODS: A cross-sectional study was conducted from February to December 2008; women with age of 18 to 45 years, with low risk for sexually transmitted disease (STD) were invited to participate. Blood samples were collected in filter paper, and HTLV-1/2 immunoenzymatic test (ELISA) was performed as a screening test. Women with reactive results were submitted to peripheral venous blood collection for ELISA repetition, followed by Western blot (WB) and real-time PCR to confirm and discriminate the infection between virus types 1 and 2. RESULTS: Of the 2,044 women tested, seven (0.3%) were ELISA reactive and confirmed positive (four were HTLV-1, and three were HTLV-2). All positive women were oriented not to breastfeed their newborns. CONCLUSIONS: This study showed that the virus is present in high prevalence in that population. Further studies covering other segments of the population are necessary to better characterize the presence of HTLV-1/2 in Maranhão and to elicit measures to prevent its spread.

Plasmodium vivax malaria: related factors to severity in the State of Maranhão, Brazil

INTRODUCTION: Malaria caused by Plasmodium vivax species has shown signs of severity, recorded with increasing frequency in the medical literature. This study aimed to characterize the signs of severe malaria by Plasmodium vivax in the State of Maranhão, Brazil. METHODS: A descriptive cohort study of patients assisted in the field and a historical and concurrent study of a series of cases among hospitalized patients were undertaken to identify the clinical and laboratory signs of severity. RESULTS: A total of 153 patients were included in the study, 13 of whom were hospitalized. Males made up the majority, numbering 103 (67.3%). The age of the patients ranged from 10 to 70 years, 92.2% were natives of the State of Maranhão, and 65% of the patients had had malaria before. The average time elapsed between symptom onset and diagnosis among outpatients was three days, while among hospitalized patients this average reached 15.5 days, a statistically significant difference (p=0.001). The parasitemia ranged from 500 to 10,000 parasites/µl in 92.8% of cases. The clinical and laboratory manifestations of severity were vomiting and diarrhea, jaundice, drowsiness, mental confusion, seizures, loss of consciousness, agitation, bleeding, pale skin, coughing and dyspnea, thrombocytopenia, anemia, elevation of nitrogenous compounds, and elevated transaminases and bilirubin. CONCLUSIONS: The monitoring of malaria patients with Plasmodium vivax showed the possibility of aggravation, the intensity of which varied in different circumstances, especially the interval time between falling ill and diagnostic confirmation.

The first report of infection with Klebsiella pneumoniae carrying the bla kpc gene in State of Mato Grosso do Sul, Brazil

The increased frequency and dissemination of enterobacteria resistant to various antimicrobials is currently worldwide concern. In January 2010, a 94-year-old patient with chronic lymphocytic leukemia was admitted to the University Hospital. This patient died 21 days after hospitalization due to the clinical worsening. Klebsiella pneumoniae producing of extended-spectrum β-lactamases (ESBLs) was isolated of urine culture. This bacterium demonstrated resistance to ceftazidime, ciprofloxacin, levofloxacin, ertapenem and imipenem. Susceptibility to cefoxitin, cefepime, meropenem, colistin and tigecycline. This study reports the first case of infection by Klebsiella pneumoniae carrying the bla kpc gene in the State of Mato Grosso do Sul, Brazil.

Glucocorticoid-induced tumor necrosis factor receptor expression in patients with cervical human papillomavirus infection

Introduction The progression of human papillomavirus (HPV) infection in the anogenital tract has been associated with the involvement of cells with regulatory properties. Evidence has shown that glucocorticoid-induced tumor necrosis factor receptor (GITR) is an important surface molecule for the characterization of these cells and proposes that GITR ligand may constitute a rational treatment for many cancer types. We aimed to detect the presence of GITR and CD25 in cervical stroma cells with and without pathological changes or HPV infection to better understand the immune response in the infected tissue microenvironment. Methods We subjected 49 paraffin-embedded cervical tissue samples to HPV DNA detection and histopathological analysis, and subsequently immunohistochemistry to detect GITR and CD25 in lymphocytes. Results We observed that 76.9% of all samples with high GITR expression were HPV-positive regardless of histopathological findings. High GITR expression (77.8%) was predominant in samples with ≥1,000 RLU/PCB. Of the HPV-positive samples negative for intraepithelial lesion and malignancy, 62.5% had high GITR expression. High GITR expression was observed in both carcinoma and high-grade squamous intraepithelial lesion (HSIL) samples (p = 0.16). CD25 was present in great quantities in all samples. Conclusions The predominance of high GITR expression in samples with high viral load that were classified as HSIL and carcinoma suggests that GITR+ cells can exhibit regulatory properties and may contribute to the progression of HPV-induced cervical neoplasia, emphasizing the importance of GITR as a potential target for immune therapy of cervical cancer and as a disease evolution biomarker.

Schistosomiasis mansoni in urban Northeast Brazil: influence of rainfall regime on the population dynamics of Biomphalaria sp.

Introduction Our objective was to evaluate the influence of rainfall regime on the population dynamics of Biomphalaria in a potential urban focus of schistosomiasis in Aracaju, Brazil, during 2009-2010. Methods Snails were collected monthly and were counted, measured and identified; the level of infection and fecal contamination at the sampling sites was determined; rainfall data were obtained. Results High levels of fecal contamination were observed, and the abundance of Biomphalaria glabrata increased during the rainy and post-rainy seasons. The snails' size was variable, and infected snails were identified independently of rainfall. Conclusions These results provide evidence of anthropogenic and climate interference in an urban focus of schistosomiasis in the Aracaju metropolitan area.

Dengue infection in children and adolescents: Clinical profile in a reference hospital in northeast Brazil

Introduction This study aimed to describe the clinical spectrum of dengue in children and adolescents from a hyperendemic region who were admitted for hospitalization. Methods A retrospective study was conducted on patients diagnosed with dengue infection upon admission to a reference center in Fortaleza, Brazil. Results Of the 84 patients included, 42 underwent confirmatory testing. The main symptoms were fever, abdominal pain and vomiting. The median level of serum aspartate aminotransferase was 143.5±128mg/dL. Conclusions A peculiar clinical profile was evident among children and adolescents with dengue infection in a reference center in northeast Brazil, including gastrointestinal symptoms and liver involvement.

Incidence of canine leptospirosis in the metropolitan area of Curitiba, State of Parana, Southern Brazil

Introduction The incidence of canine leptospirosis in Brazil needs to be assessed Methods The same dogs in southern Brazil were sampled over two years to determine the prevalence, incidence and association of canine leptospirosis with various risk factors. Results In 2009, the prevalence was 33 (14.4%) of 228 dogs, with a predominance of serovar Canicola (33.4%). In 2010, 90 dogs were re-evaluated (the remaining dogs were lost to deaths, address changes and donations), and the prevalence was found to be 35 (38.9%) of 90, with the predominant serovar being Icterohaemorrhagiae (51.4%). Moreover, the incidence was 26 of 90 (28.9%), and the disease was statistically associated with age (2009) and street access (2010). Conclusions Our findings revealed instability in the dog population and age to be relevant risk factors for canine leptospirosis.

Late diagnosis and HIV infection in children attending a service of specialized care for pediatric AIDS in Brazil

Introduction: This study describes the frequency of late diagnosis and HIV among children attending a pediatric AIDS clinic. Methods: Cross-sectional study in children exposed to HIV from 2005-2008. A questionnaire was given that included questions on demographics and clinical information. Results: Two-hundred twenty-one (97.8%) children were exposed to HIV during pregnancy/childbirth. A total of 193 (87.3%) children had late enrolment in the service and late access to HIV serology. The frequency of HIV was 21.3% (95% confidence interval [CI] 15.9%-26.7%). Protective factors were earlier diagnosis [odds ratio (OR)=0.17 (0.08-0.37)] and receiving complete prophylaxis [OR=0.29 (0.09-0.97)]; being born by vaginal delivery was a risk factor [OR=4.45 (1.47-13.47)]. Conclusions: There was a high frequency of late diagnosis in this patient cohort. Earlier diagnosis is an important measure for controlling HIV among children.