A comparison of the immune parameters of dogs infected with visceral leishmaniasis using Western blot and neutralization techniques

The Western blot technique was used to demonstrate the presence of antibodies in the blood of dogs that presented canine visceral leishmaniasis. This technique was used against some specific molecules present in the lysate of the promastigote form of Leshmania chagasi.Through the association of the results of the Western blot technique with the morphological alterations seen as a result of the serum neutralization technique performed in McCoy cells (which mimetizes the macrophage) it was possible to observe the role of some molecules of great relevance in determining the disease in symptomatic dogs as well as that of some other molecules associated with asymptomatic infected dogs that may become transmitters as well as differentiating them as asymptomatic resistant dogs. In the sera analyses carried out during the immunobloting a variation of 9 to 27 immunoreacting bands was observed, which were then compared using Dice's similarity coefficient. In the dendrogram constructed on the basis of the coefficient, 50% similarity was observed among the total number of reagent bands with the promastigote lysate, thus creating five groups. The main difference observed related to the clinical condition of the dogs: symptomatic and asymptomatic dogs were found in separate groups. The asymptomatic group of dogs was distributed in two different places in the dendrogram because they presented two different behavior patterns regarding the cellular morphology in the serum neutralization reaction: the presence or absence of cellular lysis. According to this analysis it is possible to evaluate the immune status and associate it with specific markers observed in the reaction found in the Western blot strips.

Comparison of direct immunofluorescence, conventional cell culture and polymerase chain reaction techniques for detecting respiratory syncytial virus in nasopharyngeal aspirates from infants

A total of 316 samples of nasopharyngeal aspirate from infants up to two years of age with acute respiratory-tract illnesses were processed for detection of respiratory syncytial virus (RSV) using three different techniques: viral isolation, direct immunofluorescence, and PCR. Of the samples, 36 (11.4%) were positive for RSV, considering the three techniques. PCR was the most sensitive technique, providing positive findings in 35/316 (11.1%) of the samples, followed by direct immunofluorescence (25/316, 7.9%) and viral isolation (20/315, 6.3%) (p < 0.001). A sample was positive by immunofluorescence and negative by PCR, and 11 (31.4%) were positive only by RT-PCR. We conclude that RT-PCR is more sensitive than IF and viral isolation to detect RSV in nasopharyngeal aspirate specimens in newborn and infants.

Temporal differences in blood meal detection from the midguts of Triatoma infestans

We used genus/species specific PCRs to determine the temporal persistence of host DNA in Triatoma infestans experimentally fed on blood from six common vertebrate species: humans, domestic dogs, guinea pigs, chickens, mice, and pigs. Twenty third or fourth instar nymphs per animal group were allowed to feed to engorgement, followed by fasting-maintenance in the insectary. At 7, 14, 21, or 28 days post-feeding, the midgut contents from five triatomines per group were tested with the respective PCR assay. DNA from all vertebrate species was detected in at least four of five study nymphs at seven and 14 days post-feeding. DNA of humans, domestic dogs, guinea pigs, pigs, and chickens were more successfully detected (80-100%) through day 21, and less successfully (20-100%) at day 28. Findings demonstrate that species-specific PCRs can consistently identify feeding sources of T. infestans within two weeks, a biologically relevant time interval.

TEMPORAL TRENDS IN THE DETECTION RATES OF HEPATITIS B IN THE SANTA CATARINA STATE, BRAZIL

Hepatitis B is a serious public health problem. The state of Santa Catarina presents areas of high endemicity. The aim of this study was to describe temporal trends in detection rates of hepatitis B in the period from 2002 to 2009 in Santa Catarina and in its regions. A time series study was carried out. Crude rates were calculated and standardized by age using the direct method. Annual variation percentages were estimated by Joinpoint regression. There were two distinct and significant trends in Santa Catarina. From 2002 to 2006 a significant increase of 5.9% per year was observed. From 2006, there was a significant decrease of 6.4% per year. In this same period the southern and far-western regions had significant increases of 15.9% and 4.6% and significant decreases of 7.5% and 4.8%, respectively. Greater Florianópolis and Northeast also showed significant increases until 2006, of 15.4% and 17.4%, respectively. In the following period, non-significant decreases of 5.8% and 9.8% respectively were observed. Foz do Rio Itajaí and Planalto Serrano showed non-significant increases up to half of the studied period of 21.1% and 12.0%, respectively and after, significant decreases of 21.5% and 18.0%, respectively. Vale do Itajaí showed a significant decrease of 9.7%; Planalto Norte showed a non-significant decrease of 0.6% and Midwest a non-significant increase of 2.7% per year, in the period from 2002 to 2009.

IDENTIFICATION OF CANINE VISCERAL LEISHMANIASIS IN A PREVIOUSLY UNAFFECTED AREA BY CONVENTIONAL DIAGNOSTIC TECHNIQUES AND CELL-BLOCK FIXATION

After the report of a second case of canine visceral leishmaniasis (CVL) in São Bento da Lagoa, Itaipuaçu, in the municipality of Maricá, Rio de Janeiro State, an epidemiological survey was carried out, through active search, totaling 145 dogs. Indirect immunofluorescence assay (IFA), enzyme-linked immunosorbent assay (ELISA), and rapid chromatographic immunoassay based on dual-path platform (DPP(r)) were used to perform the serological examinations. The parasitological diagnosis of cutaneous fragments was performed by parasitological culture, histopathology, and immunohistochemistry. In the serological assessment, 21 dogs were seropositive by IFA, 17 by ELISA, and 11 by DPP(r), with sensitivity of 66.7%, 66.7% and 50%, and specificity of 87.2%, 90.2% and 94%, respectively for each technique. The immunohistochemistry of bone marrow using the cell-block technique presented the best results, with six positive dogs found, three of which tested negative by the other parasitological techniques. Leishmania sp. was isolated by parasitological culture in three dogs. The detection of autochthonous Leishmania infantum in Itaipuaçu, and the high prevalence of seropositive dogs confirm the circulation of this parasite in the study area and alert for the risk of expansion in the State of Rio de Janeiro.

Evolução temporal da resistência in vitro do Plasmodium falciparum às drogas antimaláricas em duas áreas da amazônia brasileira com distintas características sócio-econômicas e geográficas

Avaliou-se a evolução temporal da resistência in vitro do Plasmodium falciparum às drogas cloroquina, amodiaquina, quinino e mefloquina em duas áreas com distintas características sócio-econômicas e geográficas: Lourenço, no Estado do Amapá e Paragominas no Estado do Pará. A primeira caracteriza-se por ser uma área de garimpos a céu aberto e a segunda uma área de colonização, pecuária e extrativismo de madeiras. O estudo revela alta prevalência de resistência à cloroquina nas duas áreas (79,8% em Lourenço e 68,4% em Paragominas), enquanto que para amodiaquina e quinino observamos uma certa flutuação nas respostas para essas drogas, dependendo do período em que foi avaliada, fá para mefloquina, não foram obsewadas cepas resistentes, mas uma perda da sensibilidade ao longo do período estudado.

Análise temporal da variabilidade da freqüência cardíaca no estado basal em idosos chagásicos na forma indeterminada em área endêmica

Com o objetivo de avaliar a função autonômica cardíaca em pacientes chagásicos residentes em área endêmica, foram avaliados, por meio da análise computadorizada da variabilidade da freqüência cardíaca, 28 pacientes idosos chagásicos na forma indeterminada, 28 pacientes idosos não-chagásicos e 28 adultos jovens. Todos os pacientes chagásicos realizaram eletrocardiograma, radiografia de tórax, estudo radiológico contrastado do esôfago e cólons e ecodopplercardiograma, sendo que os não-chagásicos deixaram de realizar apenas os exames contrastados. Não houve diferença estatisticamente significativa entre os grupos, quanto às dimensões sistólica e diastólica e função sistólica do ventrículo esquerdo. Não houve diferença estatisticamente significante entre os grupos, quanto à duração média do intervalo RR. Quanto à variância, desvio padrão, coeficiente de variação, e ao pNN50, houve diferença estatisticamente significante entre o grupo jovem e os idosos, mas não entre os grupos idosos. Concluímos que, no estado basal, os grupos idosos chagásicos e não-chagásicos não diferiram quanto à modulação autonômica cardíaca no domínio do tempo.

A epidemia de AIDS no Brasil, 1991-2000: descrição espaço-temporal

A incidência de AIDS foi retratada por um estudo ecológico dos municípios brasileiros, considerando as notificações entre 1991 e 2000. Observou-se aumento da incidência em mulheres e em indivíduos com baixa escolaridade. O Estado de São Paulo, com 46% das notificações registradas no País, apresentou perfil epidemiológico distinto, quando a capital foi comparada com as cidades do interior; o Estado de Pernambuco apresentou um perfil epidemiológico semelhante àquele do início da epidemia no Brasil; o Estado Santa Catarina mostrou o predomínio de casos usuários de drogas injetáveis. O aumento de notificações pela via de transmissão ignorada indica que o viés de classificação foi crescente no sistema de vigilância epidemiológica.

Comparison of the thick smear and Kato-Katz techniques for diagnosis of intestinal helminth infections

This study compared the efficiency of Kato-Katz thick smear and thick smear techniques for the diagnosis of intestinal helminths. The sensitivity of the thick smear technique was higher than that of the Kato-Katz method for the diagnosis of all helminths except Schistosoma mansoni.

Diagnosing human asymptomatic visceral leishmaniasis in an urban area of the State of Minas Gerais, using serological and molecular biology techniques

A population-based cross-sectional study was set up in Sabará country, Southeastern Brazil, to identify asymptomatic human visceral leishmaniasis in an urban area of low disease prevalence. Blood was collected on filter paper (n=1,604 inhabitants) and examined by indirect immunofluorescent test, enzyme-linked immunosorbent assay and immunochromatographic strip test. The prevalence rates of infection ranged from 2.4 to 5.6% depending on the test used. One year later, venous blood was collected in a subset of 226 participants (102 seropositive and 124 seronegative). The tests performed were IFAT, ELISA, rk39-ELISA, polymerase chain reaction and hybridization with Leishmania donovani complex probe. No clinical signs or symptoms of leishmaniasis were observed. Using hybridization as a reference test, the sensitivity and specificity of serology were respectively: 24.8 and 71% (ELISA); 26.3 and 76.3% (rk-39); 30.1 and 63.4% (IFAT). Due to disagreements, different criteria were tested to define the infection and hybridization should be considered in epidemiological studies.

Analysis of molecular biology techniques for the diagnosis of human papillomavirus infection and cervical cancer prevention

The objective of the present study was to evaluate the usefulness of molecular methodologies to access human papillomavirus genome in the genital tract. Samples from 136 women aged 17 to 52 years old obtained from the Dr. Sérgio Franco Laboratories between 2000 and 2001, were analyzed by the hybrid capture assay and amplified by PCR with generic primers MY09/MY11 and specific primers for types 16, 18, 31, 33, 35, 58. Viral genome was detected in 71.3% of the samples by hybrid capture and 75% by amplification. When cytopathology was used as a reference method for screening lesions, hybrid capture (p=0) and amplification (p=0.002) presented positive association. The 3 methods showed absolute agreement when cytopathology confirmed papillomavirus infection and high grade intraepithelial lesion. Disagreements occurred for 10 cases: seven inflammatory cases positive by PCR and negative for hybrid capture and 3 low squamous intraepithelial lesions positive for hybrid capture but negative for amplification. In conclusion, hybrid capture was shown to be sensitive and specific enough for use in clinical routines. Moreover, the evaluation of viral load values obtained by this method were shown to be related to the severity of the lesion and merit further studies to analyze the possible association with risk of progression to malignancy.

Tendência temporal do tétano acidental no período de 1981 a 2004 em Pernambuco com avaliação do impacto da assistência em unidade de terapia intensiva sobre a letalidade

Apesar da redução na incidência de tétano acidental no Brasil, não houve queda significativa na letalidade. Nesta série de casos, comparamos a letalidade antes e após o estabelecimento padrão de manejo em unidade de terapia intensiva do paciente com tétano no Hospital Universitário Oswaldo Cruz, no período de 1981 a 2004. Em 24 anos, foram internados 1.971 pacientes e antes do manejo em Unidade de terapia intensiva a letalidade era de 35%. Durante 1997 foi instituída a unidade de terapia intensiva para assistência dos pacientes com tétano, e de 1998 a 2004, a letalidade caiu para 12,6%, OR= 0,27 (IC95%= 0,18- 0,39); p<0,001. Esta tendência foi evidenciada em todas as faixas etárias e em ambos os sexos. A centralização da assistência a esses pacientes em um único serviço especializado com Unidade de terapia intensiva de forma precoce, portanto, tem sido decisiva na redução da letalidade, por contar com a vasta experiência da equipe de saúde no manejo do tétano e melhor tratamento sintomático, antecipando as graves complicações da doença.

A epidemia de aids no Estado de São Paulo: uma aplicação do modelo espaço-temporal bayesiano completo

O Estado de São Paulo, por compreender aproximadamente 40% dos casos de aids notificados no Brasil, oferece uma situação propícia para análise espaço-temporal, visando melhor compreensão da disseminação do HIV/aids. Utilizando os casos de aids notificados ao Ministério da Saúde nos anos de 1990 a 2004 para pessoas com idade igual ou superior a 15 anos, tendo como fonte de informação o Sistema de Informação de Agravos e Notificação, Ministério da Saúde, foram estimados os riscos relativos de aids segundo sexo para períodos de 3 anos utilizando modelos bayesianos completos. Os modelos utilizados se mostraram adequados para explicar o processo de disseminação da aids no Estado de São Paulo e evidenciam os processos de feminização e interiorização da doença, além de sugerir que os municípios atualmente mais atingidos se encontram em regiões de pólos de crescimento econômico e possuem população inferior a 50.000 habitantes.