Bartonella spp. and Coxiella burnetii Associated with Community-Acquired, Culture-Negative Endocarditis, Brazil.

We evaluated culture-negative, community-acquired endocarditis by using indirect immunofluorescent assays and molecular analyses for Bartonella spp. and Coxiella burnetii and found a prevalence of 19.6% and 7.8%, respectively. Our findings reinforce the need to study these organisms in patients with culture-negative, community-acquired endocarditis, especially B. henselae in cat owners.

Febre amarela silvestre: reemergência de transmissão no estado de São Paulo, Brasil, 2009* / Yellow fever: reemerging in the state of São Paulo, Brazil, 2009

Descrever a investigação do surto de febre amarela silvestre e as principais medidas de controle realizadas no estado de São Paulo. MÉTODOS: Estudo descritivo do surto de febre amarela silvestre na região sudoeste do estado, entre fevereiro e abril de 2009. Foram avaliados casos suspeitos e confirmados em humanos e primatas não humanos. A investigação entomológica, em ambiente silvestre, envolveu captura em solo e copa de árvore para identificação das espécies e detecção de infecção natural. Foram realizadas ações de controle de Aedes aegypti em áreas urbanas. A vacinação foi direcionada para residentes dos municípios com confirmação de circulação viral e nos municípios contíguos, conforme recomendação nacional. RESULTADOS: Foram confirmados 28 casos humanos (letalidade 39,3%) em áreas rurais de Sarutaiá, Piraju, Tejupá, Avaré e Buri. Foram notificadas 56 mortes de primatas não humanos, 91,4% do gênero Alouatta sp. A epizootia foi confirmada laboratorialmente em dois primatas não humanos, sendo um em Buri e outro em Itapetininga. Foram coletados 1.782 mosquitos, entre eles Haemagogus leucocelaenus, Hg. janthinomys/capricornii, Sabethes chloropterus, Sa. purpureus e Sa. undosus. O vírus da febre amarela foi isolado de um lote de Hg. leucocelaenus procedente de Buri. A vacinação foi realizada em 49 municípios, com 1.018.705 doses aplicadas e o registro de nove eventos adversos graves pós-vacinação. CONCLUSÕES: Os casos humanos ocorreram entre fevereiro e abril de 2009 em áreas sem registro de circulação do vírus da febre amarela há mais de 60 anos. A região encontrava-se fora da área com recomendação de vacinação, com alto percentual da população suscetível. A adoção oportuna de medidas de controle permitiu a interrupção da transmissão humana em um mês, assim como a confirmação da circulação viral em humanos, primatas não humanos e mosquitos. Os isolamentos facilitaram a identificação das áreas de circulação viral, mas são importantes novos estudos para esclarecer a dinâmica de transmissão da doença...

Immunopathogenesis of dengue hemorrhagic fever: contribution to the study of human liver lesions.

Dengue infection is an important tropical disease worldwide. The host immune response has been studied in order to better understand lesion mechanisms. It was performed an immunohistochemical study in 14 specimens of liver from patients with dengue hemorrhagic fever (DHF) to characterize cytokines and some factors present in liver lesions and their possible role in the pathogenesis of hepatic injury. Portal tract and hepatic acinus presented high expression of TLR2, TLR3, IL6, and granzyme B. Hepatic acinus also presented iNOS, IL18, and TGF-beta. Cells expressing IL12, IL13, JAk1, STAT1, and NF-κB were rarely visualized. Treg cells foxp3+ were absent. TLR2 and TLR3 seem to participate in cellular activation and cytokine production. Cytotoxic response seems to play a role. Although TGF-beta promotes the activation of Foxp3+ regulatory T cells, IL6 can significantly suppresses their generation. The expression of Treg cells is diminished probably as a result of the high frequency of these cytokines. Both cytokines play a role in the increased vascular permeability and edema observed in dengue liver specimens, with consequent plasma leakage and severity of the disease. It was observed a regular expression of IL-18 in hepatocytes and lymphocytes of the inflammatory infiltrate in portal tract, which reflects the acute inflammatory response that occurs in the liver and contributes to hepatic injury. At least in part, the increased number of cells expressing IL-18 could play a role of "up" regulation of FasL and correlate to the phenomenon of apoptosis, a mechanism of destruction of hepatocytes in DHF.

Febre amarela silvestre: reemergencia de transmissao no estado de Sao Paulo, Brasil, 2009 / Fiebre amarilla silvestre: re emergencia de transmision en el estado de Sao Paulo, Brasil, 2009 / Yellow fever: reemerging in the state of Sao Paulo, Brazil, 2009

OBJETIVO Descrever a investigação do surto de febre amarela silvestre e as principais medidas de controle realizadas no estado de São Paulo. MÉTODOS Estudo descritivo do surto de febre amarela silvestre na região sudoeste do estado, entre fevereiro e abril de 2009. Foram avaliados casos suspeitos e confirmados em humanos e primatas não humanos. A investigação entomológica, em ambiente silvestre, envolveu captura em solo e copa de árvore para identificação das espécies e detecção de infecção natural. Foram realizadas ações de controle de Aedes aegypti em áreas urbanas. A vacinação foi direcionada para residentes dos municípios com confirmação de circulação viral e nos municípios contíguos, conforme recomendação nacional. RESULTADOS Foram confirmados 28 casos humanos (letalidade 39,3%) em áreas rurais de Sarutaiá, Piraju, Tejupá, Avaré e Buri. Foram notificadas 56 mortes de primatas não humanos, 91,4% do gênero Alouatta sp . A epizootia foi confirmada laboratorialmente em dois primatas não humanos, sendo um em Buri e outro em Itapetininga. Foram coletados 1.782 mosquitos, entre eles Haemagogus leucocelaenus , Hg. janthinomys/capricornii , Sabethes chloropterus , Sa. purpureus e Sa. undosus . O vírus da febre amarela foi isolado de um lote de Hg. leucocelaenus procedente de Buri. A vacinação foi realizada em 49 municípios, com 1.018.705 doses aplicadas e o registro de nove eventos adversos graves pós-vacinação. CONCLUSÕES Os casos humanos ocorreram entre fevereiro e abril de 2009 em áreas sem registro de circulação do vírus da febre amarela há mais de 60 anos. A região encontrava-se fora da área com recomendação de vacinação, com alto percentual da p...

OBJETIVO Describir la investigación de brote de fiebre amarilla silvestre y las principales medidas de control realizadas en el estado de Sao Paulo. MÉTODOS Estudio descriptivo del brote de fiebre amarilla silvestre en la región suroeste del Estado, entre febrero y abril de 2009. Se evaluaron casos sospechosos y confirmados en humanos y primates no humanos. La investigación entomológica, en ambiente silvestre, involucró capturo en suelo y copa de árboles para identificación de las especies y detección de infección natural. Se realizaron acciones de control de Aedes aegypti en áreas urbanas. La vacunación fue direccionada a residentes de los municipios con confirmación de circulación viral y en los municipios contiguos, siguiendo recomendación nacional. RESULTADOS Se confirmaron 28 casos en humanos (letalidad 39,3%) en áreas rurales de Sarutaiá, Pirajú, Tejupá, Avaré y Buri. Se notificaron 56 muertes de primates no humanos, 91,4% del género Allouatta sp. La epizootia fue confirmada laboratorialmente en dos primates no humanos siendo uno de Buri y el otro de Itapetininga. Se colectaron 1.782 mosquitos, entre ellos Haemagogus leucocelaenus, Hg. janthinomys/capricornii, y Sabethes chloropterus, Sa. purpureus y Sa. undosus. El virus de la fiebre amarilla fue aislado de un lote de Hg. leucocelaenus procedente de Buri. La vacunación fue realizada en 49 municipios, con 1.018.705 dosis aplicadas y el registro de nueve eventos adversos graves post-vacunación. CONCLUSIONES Los casos humanos ocurrieron entre febrero a abril de 2009 en áreas sin registro de circulación del virus de la fiebre amarilla por más de 60 años. La región se encontraba fuera del área de recomendación de vacunación, con alto porcentaje de población susceptible. La adopción oportuna de medidas de control permitió ...

OBJECTIVE To describe the investigation of a sylvatic yellow fever outbreak in the state of Sao Paulo and the main control measures undertaken. METHODS This is a descriptive study of a sylvatic yellow fever outbreak in the Southwestern region of the state from February to April 2009. Suspected and confirmed cases in humans and in non-human primates were evaluated. Entomological investigation in sylvatic environment involved capture at ground level and in the tree canopy to identify species and detect natural infections. Control measures were performed in urban areas to control Aedes aegypti . Vaccination was directed at residents living in areas with confirmed viral circulation and also at nearby cities according to national recommendation. RESULTS Twenty-eight human cases were confirmed (39.3% case fatality rate) in rural areas of Sarutaiá, Piraju, Tejupá, Avaré and Buri. The deaths of 56 non-human primates were also reported, 91.4% were Allouatta sp. Epizootics was confirmed in two non-human primates in the cities of Itapetininga and Buri. A total of 1,782 mosquitoes were collected, including Haemagogus leucocelaenus , Hg. janthinomys/capricornii , and Sabethes chloropterus, Sa. purpureus and Sa. undosus . Yellow fever virus was isolated from a group of Hg. Leucocelaenus from Buri. Vaccination was carried out in 49 cities, with a total of 1,018,705 doses. Nine serious post-vaccination adverse events were reported. CONCLUSIONS The cases occurred between February and April 2009 in areas with no recorded yellow fever virus circulation in over 60 years. The outbreak region occurred outside the original recommended vaccination area with a high percentage of susceptible population. The fast adoption of control measures interrupted the human transmission within a month and the confirmation of viral circulation in humans, monkeys and mosquitoes. The results allowed the identification of new areas of viral circulation but ...

Yellow fever epizootics in non-human primates, São Paulo state, Brazil, 2008-2009 / Epizootias de febre amarela em primatas não humanos no estado de São Paulo, Brasil, 2008-2009

Since 2000, the expansion of Sylvatic Yellow Fever (YF) has been observed in the southeast of Brazil, being detected in areas considered silent for decades. Epizootics in non-human primates (NHPs) are considered sentinel events for the detection of human cases. It is important to report epizootic events that could have impact on the conservation status of susceptible species. We describe the epizootics in NHPs, notified in state of São Paulo, Brazil, between September 2008 to August 2009. Ninety-one epizootic events, involving 147 animals, were reported in 36 counties. Samples were obtained from 65 animals (44.2%). Most of the epizootics (46.6%) were reported between March and April, the same period during which human cases of YF occurred in the state. Biological samples were collected from animals found dead and were sent to Instituto Adolfo Lutz, in São Paulo. Two samples, collected in two counties without an indication for YF vaccination, were positive for the virus. Another 48 animals were associated with YF by clinical-epidemiological linkage with laboratory confirmed cases. Because the disease in human and NHPs occurred in the same period, the detection of the virus in NHPs did not work as sentinel, but aided in the delineation of new areas of risk.

Desde 2000, vem sendo observada a expansão da febre amarela (FA) no Sudeste do Brasil, sendo detectados casos em áreas consideradas silenciosas por décadas. Epizootias em primatas não humanos (NHPs) são considerados eventos sentinela para a detecção de casos humanos. É importante relatar eventos epizoóticos que podem ter impacto sobre o estado de conservação de espécies sensíveis. Descrevemos as epizootias, notificadas em NHPs no estado de São Paulo, Brasil, entre setembro de 2008 a agosto de 2009. Noventa e um eventos epizoóticos, envolvendo 147 animais, foram notificados em 36 municípios. As amostras foram obtidas a partir de 65 animais (44,2%). A maioria das epizootias (46,6%) foram registradas entre março e abril, no mesmo período no qual YF em que casos humanos ocorreram no estado. As amostras biológicas foram coletadas de animais encontrados mortos e enviadas ao Instituto Adolfo Lutz, em São Paulo. Duas amostras, coletadas em dois municípios, sem indicação para a vacinação de febre amarela, foram positivos para o vírus. Outros 48 animais foram associados com FA por vínculo clínico-epidemiológico com casos confirmados laboratorialmente. Devido a doença em humanos e NHPs terem ocorrido no mesmo período, a detecção do vírus em NHPs não funcionou como sentinela, mas ajudou no processo de delimitação de novas áreas de risco.

Yellow fever epizootics in non-human primates, São Paulo state, Brazil, 2008-2009.

Since 2000, the expansion of Sylvatic Yellow Fever (YF) has been observed in the southeast of Brazil, being detected in areas considered silent for decades. Epizootics in non-human primates (NHPs) are considered sentinel events for the detection of human cases. It is important to report epizootic events that could have impact on the conservation status of susceptible species. We describe the epizootics in NHPs, notified in state of São Paulo, Brazil, between September 2008 to August 2009. Ninety-one epizootic events, involving 147 animals, were reported in 36 counties. Samples were obtained from 65 animals (44.2%). Most of the epizootics (46.6%) were reported between March and April, the same period during which human cases of YF occurred in the state. Biological samples were collected from animals found dead and were sent to Instituto Adolfo Lutz, in São Paulo. Two samples, collected in two counties without an indication for YF vaccination, were positive for the virus. Another 48 animals were associated with YF by clinical-epidemiological linkage with laboratory confirmed cases. Because the disease in human and NHPs occurred in the same period, the detection of the virus in NHPs did not work as sentinel, but aided in the delineation of new areas of risk.

Disseminated amphotericin-resistant fusariosis in acute leukemia patients: report of two cases.

Disseminated fusariosis has emerged as a significant, usually fatal infection in immunocompromised hosts despite antifungal treatment. We describe here two patients with acute leukemia who developed disseminated amphotericin-resistant fusariosis, and review of six studies of cases series in the literature. Two Fusarium solani strains were isolated from blood and skin cultures of one patient, and one strain from the blood culture of the second patient. Both patients died despite antifungal treatment. Strains were identified by sequencing of ITS1 and ITS4 regions. Random amplified polymorphic DNA analysis of the three F. solani isolates showed a low degree of similarity. Screening for Fusarium spp. contaminants within our facility was negative. Using the CLSI M-38-A2 broth dilution method and E tests(®), we found that the MICs were low for voriconazole (0.12 and 0.5 mg/L, respectively), unexpectedly high for amphotericin B (≥8 and ≥32 µg/mL, respectively) and itraconazole (≥16 mg/ml). Patients with leukemia or persistent neutropenia should be assessed for disseminated fungal infections, including biopsy and skin cultures. Antifungal susceptibility tests are important due to the possibility of the strains being amphotericin resistant. Treatments must be aggressive, with high doses of antifungals or combined therapy.

[Yellow fever: reemerging in the state of Sao Paulo, Brazil, 2009]. / Febre amarela silvestre: reemergência de transmissão no estado de São Paulo, Brasil, 2009.

OBJECTIVE: To describe the investigation of a sylvatic yellow fever outbreak in the state of Sao Paulo and the main control measures undertaken. METHODS: This is a descriptive study of a sylvatic yellow fever outbreak in the Southwestern region of the state from February to April 2009. Suspected and confirmed cases in humans and in non-human primates were evaluated. Entomological investigation in sylvatic environment involved capture at ground level and in the tree canopy to identify species and detect natural infections. Control measures were performed in urban areas to control Aedes aegypti . Vaccination was directed at residents living in areas with confirmed viral circulation and also at nearby cities according to national recommendation. RESULTS: Twenty-eight human cases were confirmed (39.3% case fatality rate) in rural areas of Sarutaiá, Piraju, Tejupá, Avaré and Buri. The deaths of 56 non-human primates were also reported, 91.4% were Allouatta sp. Epizootics was confirmed in two non-human primates in the cities of Itapetininga and Buri. A total of 1,782 mosquitoes were collected, including Haemagogus leucocelaenus , Hg. janthinomys/capricornii , and Sabethes chloropterus, Sa. purpureus and Sa. undosus . Yellow fever virus was isolated from a group of Hg. Leucocelaenus from Buri. Vaccination was carried out in 49 cities, with a total of 1,018,705 doses. Nine serious post-vaccination adverse events were reported. CONCLUSIONS: The cases occurred between February and April 2009 in areas with no recorded yellow fever virus circulation in over 60 years. The outbreak region occurred outside the original recommended vaccination area with a high percentage of susceptible population. The fast adoption of control measures interrupted the human transmission within a month and the confirmation of viral circulation in humans, monkeys and mosquitoes. The results allowed the identification of new areas of viral circulation but further studies are required to clarify the dynamics of the spread of this disease.

Yellow fever outbreak affecting Alouatta populations in southern Brazil (Rio Grande do Sul State), 2008-2009.

The natural transmission cycle of Yellow Fever (YF) involves tree hole breeding mosquitoes and a wide array of nonhuman primates (NHP), including monkeys and apes. Some Neotropical monkeys (howler monkeys, genus Alouatta) develop fatal YF virus (YFV) infections similar to those reported in humans, even with minimum exposure to the infection. Epizootics in wild primates may be indicating YFV circulation, and the surveillance of such outbreaks in wildlife is an important tool to help prevent human infection. In 2001, surveillance activities successfully identified YF-related death in a black-and-gold howler monkey (Alouatta caraya), Rio Grande do Sul State (RGS) in southern Brazil, and the YFV was isolated from a species of forest-dwelling mosquito (Haemagogus leucocelaenus). These findings led the State Secretariat of Health to initiate a monitoring program for YF and other 18 arboviral infections in Alouatta monkeys. The monitoring program included monkey captures, reporting of monkey casualties by municipalities, and subsequent investigations. If monkey carcasses were found in forests, samples were collected in a standardized manner and this practice resulted in increased reporting of outbreaks. In October 2008, a single howler monkey in a northwestern RGS municipality was confirmed to have died from YF. From October 2008 to June 2009, 2,013 monkey deaths were reported (830 A. caraya and 1,183 A. guariba clamitans). Viruses isolation in blood, viscera, and/or immunohistochemistry led to the detection of YF in 204 of 297 (69%) (154 A. g. clamitans and 50 A. caraya) dead Alouatta monkeys tested. The number of municipalities with confirmed YFV circulation in howlers increased from 2 to 67 and 21 confirmed human cases occurred. This surveillance system was successful in identifying the largest YF outbreak affecting wild NHP ever recorded.

Reemergence of yellow fever: detection of transmission in the State of São Paulo, Brazil, 2008.

INTRODUCTION: Following yellow fever virus (YFV) isolation in monkeys from the São José do Rio Preto region and two fatal human autochthonous cases from the Ribeirão Preto region, State of São Paulo, Brazil, two expeditions for entomological research and eco-epidemiological evaluation were conducted. METHODS: A total of 577 samples from humans, 108 from monkeys and 3,049 mosquitoes were analyzed by one or more methods: virus isolation, ELISA-IgM, RT-PCR, histopathology and immunohistochemical. RESULTS: Of the 577 human samples, 531 were tested by ELISA-IgM, with 3 positives, and 235 were inoculated into mice and 199 in cell culture, resulting in one virus isolation. One sample was positive by histopathology and immunohistochemical. Using RT-PCR, 25 samples were processed with 4 positive reactions. A total of 108 specimens of monkeys were examined, 108 were inoculated into mice and 45 in cell culture. Four virus strains were isolated from Alouatta caraya. A total of 931 mosquitoes were captured in Sao Jose do Rio Preto and 2,118 in Ribeirão Preto and separated into batches. A single isolation of YFV was derived from a batch of 9 mosquitoes Psorophora ferox, collected in Urupês, Ribeirão Preto region. A serological survey was conducted with 128 samples from the municipalities of São Carlos, Rincão and Ribeirão Preto and 10 samples from contacts of patients from Ribeirão Preto. All samples were negative by ELISA-IgM for YFV. CONCLUSIONS: The results confirm the circulation of yellow fever, even though sporadic, in the Sao Paulo State and reinforce the importance of vaccination against yellow fever in areas considered at risk.

Reemergence of yellow fever: detection of transmission in the State of São Paulo, Brazil, 2008 / Reemergência de febre amarela: detecção de transmissão no Estado de São Paulo, Brasil, 2008

INTRODUCTION: Following yellow fever virus (YFV) isolation in monkeys from the São José do Rio Preto region and two fatal human autochthonous cases from the Ribeirão Preto region, State of São Paulo, Brazil, two expeditions for entomological research and eco-epidemiological evaluation were conducted. METHODS: A total of 577 samples from humans, 108 from monkeys and 3,049 mosquitoes were analyzed by one or more methods: virus isolation, ELISA-IgM, RT-PCR, histopathology and immunohistochemical. RESULTS: Of the 577 human samples, 531 were tested by ELISA-IgM, with 3 positives, and 235 were inoculated into mice and 199 in cell culture, resulting in one virus isolation. One sample was positive by histopathology and immunohistochemical. Using RT-PCR, 25 samples were processed with 4 positive reactions. A total of 108 specimens of monkeys were examined, 108 were inoculated into mice and 45 in cell culture. Four virus strains were isolated from Alouattacaraya. A total of 931 mosquitoes were captured in Sao Jose do Rio Preto and 2,118 in Ribeirão Preto and separated into batches. A single isolation of YFV was derived from a batch of 9 mosquitoes Psorophoraferox, collected in Urupês, Ribeirão Preto region. A serological survey was conducted with 128 samples from the municipalities of São Carlos, Rincão and Ribeirão Preto and 10 samples from contacts of patients from Ribeirão Preto. All samples were negative by ELISA-IgM for YFV. CONCLUSIONS: The results confirm the circulation of yellow fever, even though sporadic, in the Sao Paulo State and reinforce the importance of vaccination against yellow fever in areas considered at risk.

INTRODUÇÃO: A partir do isolamento do vírus febre amarela (VFA), de macacos, da região de São José do Rio Preto e de dois casos humanos autóctones fatais, da região de Ribeirão Preto, Estado de São Paulo, foram realizadas duas expedições para pesquisa entomológica e avaliação ecoepidemiológica. MÉTODOS: Um total de 577 amostras de humanos, 108 de macacos e 3.049 mosquitos foram analisados por um ou mais métodos: isolamento viral, ELISA-IgM, RT-PCR, histopatologia e imunohistoquímica. RESULTADOS: De 577 amostras humanas, 531 foram testadas por ELISA-IgM, sendo 3 positivas, 235 foram inoculadas em camundongos, 199 em cultura de células, obtendo-se 1 isolamento viral. Uma amostra foi positiva por histopatologia e imunohistoquímica. Por RT-PCR foram processadas 25 amostras com 4 reações positivas. Os 108 espécimes de macacos foram inoculados em camundongos, 45 em cultura de células, obtendo-se 4 isolamentos de VFA, de Alouatta caraya. Um total de 931 mosquitos foram capturados em São José do Rio Preto e 2.118 em Ribeirão Preto e separados em lotes. Um único isolamento de VFA foi derivado de um lote de 9 mosquitos Psorophora ferox, coletados em Urupês, região de Ribeirão Preto. Um inquérito sorológico foi realizado com 128 amostras dos municípios de São Carlos, Rincão e Ribeirão Preto e mais 10 amostras de contactantes de pacientes de Ribeirão Preto. Todas as amostras foram negativas por ELISA-IgM para VFA. CONCLUSÕES: Os resultados confirmam a circulação, mesmo que esporádica, do VFA no Estado de São Paulo e reforça a importância da vacinação antiamarílica nas áreas consideradas de risco.

Comparação da evidenciação dos miofibroblastos marcados por imuno-histoquímica em tecido de granulação quando fixado por formol ou por álcool a 70% / Comparison of myofibroblast visualization by immunohistochemical staining in granulation tissue fixed in formaldehyde or 70% alcohol

Introdução: Para o adequado estudo anatomopatológico dos tecidos obtidos dos pacientes é crucial a boa técnica para a melhor evidenciação das células pesquisadas. É imprescindível que não só a técnica ideal de coloração/marcação seja utilizada, mas também a correta técnica de fixação da peça cirúrgica. Em trabalho prévio pesquisando miofibroblastos e contração na pele restaurada de áreas doadoras de enxertos de pele de espessura parcial, verificamos baixa densidade de miofibroblastos, ao contrário dos estudos em tecido de granulação. Investigando as causas disso, e baseando-se em artigo específico sobre a eficácia da evidenciação imuno-histoquímica com diferentes métodos de fixação tecidual, comparamos a eficácia dos diferentes fixadores, formol a 10% e álcool a 70% para a evidenciação de miofibroblastos em tecido de granulação de úlceras crônicas de 21 pacientes. Método: Estudamos 42 amostras por meio de técnica imuno-histoquímica, utilizando-se o anticorpo anti-Smooth Muscle Actin, nas quais contamos os miofibrobastos em 10 campos com aumento de 400 X. Resultados: Demonstraram-se melhores resultados qualitativo e quantitativo no material fixado em álcool, quando comparado ao fixado em formol, com densidades médias de miofibroblastos de 86,5 e 48 células/mm2, respectivamente, o que nos leva a recomendar o álcool como fixador de tecidos para estas análises imuno-histoquímicas.

Background: The use of a good visualization technique is crucial for an adequate pathological anatomy study of tissues obtained from patients. Optimal staining and fixation techniques of the surgical piece are essential. In a previous study investigating myofibroblasts and contraction in split skingraft donor sites, a low density of myofibroblasts was observed, as opposed to studies in granulation studies. To investigate the causes for this finding and based on a specific manuscript on the efficacy of immunohistochemical visualization using different tissue fixation methods, we compared the efficacy of fixation in 10% formaldehyde and 70% alcohol for the visualization of myofibroblasts in granulation tissues of chronic ulcers in 21 patients. Method: Forty-two samples were studied by immunohistochemistry, using the anti-Smooth Muscle Actin antibody, and myofibroblasts were counted in 10 fields with a magnification of 400X. Results: Better qualitative and quantitative results were observed in alcohol-fixed materials, when compared to those fixed in formaldehyde, with mean myofibroblasts densities of 86.5 and 48 cells/mm2, respectively, which encourages us to recommend the use of alcohol for tissue fixation in immunohistochemical studies.

Human visceral leishmaniasis expresses Th1 pattern in situ liver lesions.

The architectural and infiltrate pattern of liver human visceral leishmaniasis (HVL) have been systematically classified as typical, fibrogenic or nodular. Despite this histopathological classification, the immune response based on cytokines and cellular phenotypes have never been performed. The aim of this study was to determine the immunophenotypic pattern and cytokine profile of the nodular involvement of the liver in HVL. We evaluated nine cases of the nodular form of HVL. In situ immune response was studied through cytokine analysis and immunohistochemical study for phenotype markers: IL-1, IL-4, IL-10, TNF-alpha, IFN-gamma, CD4+ T cells, CD8+ T cells, CD20, CD68, CD57 and macrophage activation was determined by evaluation of iNOS activity. HVL seems to be related to a better immune response. Amastigotes were rarely found on liver sections. Leishmania antigen expression was also rare and located in the inflammatory nodules. The lower expression of IL-4 and IL-10, moderate expression of TNF-alpha and IFN-gamma demonstrate a panorama of Th1 phenotype. The increased expression of NK cells could help in sustaining this model of response. This pattern of immune response is probably responsible for improvement in the parasite's clearance from liver tissue and it is a prognostic marker of human visceral leishmaniasis.

Isolated lymphadenitis due to Histoplasma capsulatum diagnosed by fine-needle aspiration biopsy and immunohistochemistry.

In the disseminated form of histoplasmosis, isolation and further identification of Histoplasma capsulatum can be performed by several methods, namely, bone marrow aspiration, blood culture, and liver biopsy. Lymph node disease usually is diagnosed by excisional biopsy. Although fungal stains can identify this fungus, detection of specific antigens by immunohistochemistry shows a higher specificity and sensitivity. This approach can use the cell block method when the material is not sent to fungal cultures or fresh staining.

Infectious diseases in paediatric pathology: experience from a developing country.

Infectious and parasitic diseases have always challenged man. Although many of them are typically seen in some areas of the world and can be adequately managed by just improving socioeconomic status and sanitary conditions, they are still quite prevalent and may sometimes be seen outside their original geographical areas. Human migration due to different reasons, tourism, blood transfusion and solid organ transplantation has created new concerns for health professionals all over the world. If not for diagnostic purposes, at least these tropical and infectious diseases should be largely known because their epidemiology, pathogenesis, host/parasite interaction, inflammatory and reparative responses are quite interesting and teach us about human biology. Curiosity is inherent to pathology practice and so we are compelled to look for things other than tumours or degenerative diseases. This review focuses on infectious and parasitic diseases found in a developing country and brings up-to-date information on diseases caused by viruses (dengue, yellow fever), bacteria (typhoid fever, leprosy), parasites (Chagas' disease, cutaneous and visceral leishmaniasis, amoebiasis, Capillaria hepatica, schistosomiasis, cysticercosis) and caused by fungi (paracoccidioidomycosis, cryptococcosis, histoplasmosis) that may be useful for pathologists when facing somewhat strange cases from developing countries.

Infectious diseases in paediatric pathology: experience from a developing country

Infectious and parasitic diseases have always challenged man. Although many of them are typically seen in some areas of the world and can be adequately managed by just improving socioeconomic status and sanitary conditions, they are still quite prevalent and may sometimes be seen outside their original geographical areas. Human migration due to different reasons, tourism, blood transfusion and solid organ transplantation has created new concerns for health professionals all over the world. If not for diagnostic purposes, at least these tropical and infectious diseases should be largely known because their epidemiology, pathogenesis, host/parasite interaction, inflammatory and reparative responses are quite interesting and teach us about human biology. Curiosity is inherent to pathology practice and so we are compelled to look for things other than tumours or degenerative diseases. This review focuses on infectious and parasitic diseases found in a developing country and brings up-to-date information on diseases caused by viruses (dengue, yellow fever), bacteria (typhoid fever, leprosy), parasites (Chagas' disease, cutaneous and visceral leishmaniasis, amoebiasis, Capillaria hepatica, schistosomiasis, cysticercosis) and caused by fungi (paracoccidioidomycosis, cryptococcosis, histoplasmosis) that may be useful for pathologists when facing somewhat strange cases from developing countries.

Portal CD4+ and CD8+ T lymphocyte correlate to intensity of interface hepatitis in chronic hepatitis C.

BACKGROUND: The pathogenesis of chronic hepatitis C is still a matter of debate. CD4+ and CD8+ T lymphocytes (TL) are typically observed within the portal and periportal spaces of affected livers, but their functional role in hepatitis C progression has not been fully elucidated. METHODS: CD4+ and CD8+ TL were quantified by immunohistochemistry in portal and periportal spaces of 39 liver biopsies from patients with chronic hepatitis C. They were associated to demographic data, histological parameters, laboratory findings of patients and hepatitis C genotypes. RESULTS: There was high numbers of CD4+ and CD8+ TL from which the density of CD4+ T was higher than CD8+ TL in portal and periportal spaces. CD4+ and CD8+ TL were directly correlated to intensity of interface hepatitis. CD8+ TL correlated to serum enzyme levels. CONCLUSION: The high numbers of CD4+ and CD8+ TL in portal and periportal spaces and their correlation to interface hepatitis suggest that hepatitis C evolution depends on the action of intrahepatic T lymphocytes, lending support to the notion of an immune-mediated mechanism in the pathogenesis of chronic hepatitis C.

Immunohistochemical study of the cellular immune response in human Pneumocystis carinii pneumonia

OBJETIVO: Trabalhos experimentais demonstram que as defesas do hospedeiro frente ao Pneumocystis carinii incluem interações complexas entre as células imunes, principalmente linfócitos TCD4+ e macrófagos alveolares. Sendo esse um agente importante associado às imunodeficiências, nosso objetivo foi caracterizar a resposta inflamatória em pulmão de necrópsias de pacientes com AIDS. MÉTODOS: Foram selecionadas 25 necrópsias com diagnóstico de pneumonia por Pneumocystis carinii para pesquisa imuno-histoquímica de linfócitos TCD4+, TCD8+, macrófagos CD68+, células NK CD57+ e células com expressão de TNF-alfa. As células imunomarcadas foram quantificadas e analisadas estatisticamente. RESULTADOS: Todos os espécimes evidenciaram elevado parasitismo na luz dos alvéolos. Observou-se espessamento septal com infiltrado inflamatório constituído predominantemente por linfócitos e macrófagos. Houve diminuição no número de linfócitos TCD4+ e aumento de linfócitos TCD8+, macrófagos e células NK. No septo alveolar freqüentemente observaram-se células expressando TNF-alfa. CONCLUSÕES: A imunossupressão relacionada à AIDS induz a diminuição de linfócitos TCD4+ e favorece a permanência de elevado parasitismo. Os componentes celulares que caracterizam o infiltrado inflamatório contribuem para os danos irreversíveis no pulmão desses pacientes.

Caracterização do fenótipo da resposta inflamatória, da expressão de citocinas e do mecanismo de morte celular no fígado de pacientes com febre hemorrágica da dengue / Caracterization of phenotype of the inflammatory reply, expression of citokines and the mechanism of cellular death in the liver of patients with dengue hemorrhagic fever

Para caracterizar a Febre Hemorrágica da Dengue no fígado através da resposta inflamatória, de citocinas e a morte celular dos hepatócitos foram analisados 17 casos de óbito no Brasil de 1999 a 2001, utilizando-se imuno-histoquímica e microscopia eletrônica. Foi observado que as lesões, o infiltrado inflamatório e a expressão de citocinas na FHD localizam-se em zona acinar 2 e o principal mecanismo de morte celular é a apoptose / To characterize the dengue hemorrhagic fever in the liver it were analyzed the inflammatory response, cytokines and the cellular death of hepatocytes by immunohistochemistry and electronic microscopy in 17 cases of death in Brazil from 1999 to 2001. It was observed that the injuries, inflammatory infiltrated and the expression of cytokines were localized in the midzone and the main mechanism of cellular death...