Immunopathological characterization of human cutaneous leishmaniasis lesions caused by Leishmania (Viannia) spp. in Amazonian Brazil.

American cutaneous leishmaniasis (ACL) is a chronic infectious disease caused by different protozoan species of Leishmania, and it is endemic in both tropical and subtropical countries. Using immunohistochemistry, we investigate the density of CD68+, lysozyme+, CD1a+, factor XIIIa+, CD4+, CD8+, CD56+, interferon (IFN)-γ+, and inducible NO synthase (iNOS+) cells. These cells were analyzed from 22 biopsy samples obtained from the lesions of ACL patients, whose infection was caused by Leishmania (Viannia) spp. Histopathological analysis showed dense mononuclear inflammatory infiltration in the dermis, which was composed of lymphocytes, macrophages, plasma cells, and discrete tissue parasitism. Granulomatous reactions were also present in the majority of cases. The density of the activated macrophages was higher than that of inactivated macrophages in the lesions. The density of Langerhans cells (CD1a+) was lower than that of dermal dendrocytes (factor XIIIa+). The density of CD8+ T lymphocytes was higher than that of CD4+ T lymphocytes. The cellular density of these immunological markers in relation to the species of Leishmania demonstrated that L. (Viannia) sp. lesions had higher IFN-γ expression than that Leishmania (Viania) braziliensis lesions. The evaluation of these markers, according to disease progression, did not reveal any significant differences. L. (Viannia) sp. infection leads to a favorable immune response in the host, as predominantly represented by lysozyme+, factor XIIIa+, CD8+ T cells, and the expression of (IFN)-γ+ at the lesion site.

Severe visceral leishmaniasis in children: the relationship between cytokine patterns and clinical features

Introduction The relationship between severe clinical manifestations of visceral leishmaniasis (VL) and immune response profiles has not yet been clarified, despite numerous studies on the subject. This study aimed to investigate the relationship between cytokine profiles and the presence of immunological markers associated with clinical manifestations and, particularly, signs of severity, as defined in a protocol drafted by the Ministry of Health (Brazil). Methods We conducted a prospective, descriptive study between May 2008 and December 2009. This study was based on an assessment of all pediatric patients with VL who were observed in a reference hospital in Maranhão. Results Among 27 children, 55.5% presented with more than one sign of severity or warning sign. Patients without signs of severity or warning signs and patients with only one warning sign had the highest interferon-gamma (IFN-γ) levels, although their interleukin 10 (IL-10) levels were also elevated. In contrast, patients with the features of severe disease had the lowest IFN-γ levels. Three patients who presented with more than two signs of severe disease died; these patients had undetectable interleukin 2 (IL-2) and IFN-γ levels and low IL-10 levels, which varied between 0 and 36.8pg/mL. Conclusions Our results showed that disease severity was associated with low IFN-γ levels and elevated IL-10 levels. However, further studies with larger samples are needed to better characterize the relationship between disease severity and cytokine levels, with the aim of identifying immunological markers of active-disease severity. .

Molecular markers of mucosa harboring gastric adenomas.

CONTEXT: Gastric adenoma is a precursor lesion of the adenocarcinoma. OBJECTIVE: To characterize gastric adenomas according to the mucin immunoexpression and to evaluate the immunoexpression of p53, p16ink4a, BCL-2, cyclin D, Ki-67, in the adenoma and in the gastric mucosa harboring adenoma. METHODS: Forty gastric specimens from 20 patients were classified as intestinal (MUC2-goblet cell mucin) or foveolar (MUC5AC-gastric-foveolar mucin) adenomas. Immunohistochemistry was performed using streptavidin-biotin-complex method. RESULTS: Twelve (60%) patients were men. The mean age was 67.9±12.9 years-old. Intestinal adenomas were detected in 13 (65%) patients and gastric type in 7 (35%). Low-grade dysplasia was present in 13 (65%) of the adenomas, high-grade in 3 (15%), and adenocarcinoma within the polyp in 4 (20%). Six (30%) patients had synchronous adenocarcinoma. p53 immunoexpression was observed in 6/20 (30%) of adenomas, and in 2/6 (33.3%) of synchronous tumors. There was an association between p53 immunoexpression and intestinal type of adenoma/tumor, P=0.04. There was no association between p16ink4a, Bcl-2, cyclin D and Ki-67 and adenoma clinicopathological characteristics. CONCLUSION: Immunohistochemistry may be useful to classify the adenomas subtypes and may define the pathway of adenoma to carcinoma sequence.

Molecular markess of mucosa harboring gastric adenoass

Context Gastric adenoma is a precursor lesion of the adenocarcinoma. Objective To characterize gastric adenomas according to the mucin immunoexpression and to evaluate the immunoexpression of p53, p16ink4a, BCL-2, cyclin D, Ki-67, in the adenoma and in the gastric mucosa harboring adenoma. Methods Forty gastric specimens from 20 patients were classified as intestinal (MUC2 - goblet cell mucin) or foveolar (MUC5AC - gastric-foveolar mucin) adenomas. Immunohistochemistry was performed using streptavidin-biotin-complex method. Results Twelve (60%) patients were men. The mean age was 67.9 ± 12.9 years-old. Intestinal adenomas were detected in 13 (65%) patients and gastric type in 7 (35%). Low-grade dysplasia was present in 13 (65%) of the adenomas, high-grade in 3 (15%), and adenocarcinoma within the polyp in 4 (20%). Six (30%) patients had synchronous adenocarcinoma. p53 immunoexpression was observed in 6/20 (30%) of adenomas, and in 2/6 (33.3%) of synchronous tumors. There was an association between p53 immunoexpression and intestinal type of adenoma/tumor, P = 0.04. There was no association between p16ink4a, Bcl-2, cyclin D and Ki-67 and adenoma clinicopathological characteristics. Conclusion Immunohistochemistry may be useful to classify the adenomas subtypes and may define the pathway of adenoma to carcinoma sequence. .

Contexto Adenoma gástrico é uma lesão precursora do adenocarcinoma. Objetivo Melhor caracterizar os adenomas de acordo com a imunoexpressão de mucinas e avaliar a imunoexpressão de p53, p16ink4a, BCL-2, cyclin D, Ki-67, nos adenomas e na mucosa gástrica adjacente. Métodos Quarenta espécimes gástricos provenientes de 20 pacientes portadores de adenomas foram classificados como do tipo intestinal (MUC2 – mucina presente nas células caliciformes) ou gástrico (MUC5AC – mucinas de padrão foveolar). Realizou-se imunoistoquímica para p53, p16ink4a, BCL-2, cyclin D e Ki-67 pelo método do complexo da estreptavidina-biotina. Resultados Doze (60%) pacientes eram homens e a média de idade foi de 67,9 ± 12,9 anos. Os adenomas foram classificados como do tipo intestinal em 13 (65%) pacientes e do tipo gástrico em 7 (35%). Displasia (neoplasia intraepitelial) de baixo grau estava presente em 13 (65%), displasia de alto grau em 3 (15%), e adenocarcinoma no pólipo adenomatoso em 4 (20%) pacientes. Observou-se immunoexpressão do p53 em 6/20 (30%) adenomas, e em 2/6 (33,3%) dos tumores sincrônicos. Houve associação entre imunoexpressão do p53 e adenoma/tumor tipo intestinal, P = 0.04. Não houve associação entre imunoexpressão do p16ink4a, Bcl-2, ciclina D e Ki-67 e as características clinicopatológicas dos adenomas. Conclusão Imunoistoquímica pode ser utilizada para caracterizar os subtipos de adenoma e talvez indicar o caminho de carcinogênese. .

Susceptibility of peritoneal macrophage from different species of neotropical primates to ex vivo Leishmania (L.) infantum chagasi-infection.

This study examined the susceptibility of peritoneal macrophage (PM) from the Neotropical primates: Callithrix jacchus, Callithrix penicillata, Saimiri sciureus, Aotus azarae infulatus and Callimico goeldii to ex vivo Leishmania (L.) infantum chagasi-infection, the etiological agent of American visceral leishmaniasis (AVL), as a screening assay for evaluating the potential of these non-human primates as experimental models for studying AVL. The PM-susceptibility to infection was accessed by the PM-infection index (PMI) at 24, 72 h and by the mean of these rates (FPMI), as well as by the TNF-α, IL-12 (Capture ELISA) and Nitric oxide (NO) responses (Griess method). At 24h, the PMI of A. azarae infulatus (128) was higher than those of C. penicillata (83), C. goeldii (78), S. sciureus (77) and C. jacchus (55). At 72h, there was a significant PMI decrease in four monkeys: A. azarae infulatus (128/37), C. penicillata (83/38), S. sciureus (77/38) and C. jacchus (55/12), with exception of C. goeldii (78/54). The FPMI of A. azarae infulatus (82.5) and C. goeldii (66) were higher than C. jacchus (33.5), but not higher than those of C. penicillata (60.5) and S. sciureus (57.5). The TNF-a response was more regular in those four primates which decreased their PMI at 24/72 h: C. jacchus (145/122 pg/mL), C. penicillata (154/130 pg/mL), S. sciureus (164/104 pg/mL) and A. azarae infulatus (154/104 pg/mL), with exception of C. goeldii (38/83 pg/mL). The IL-12 response was mainly prominent in A. infulatus and C. goeldii which presented the highest FPMI and, the NO response was higher in C. goeldii, mainly at 72 h. These findings strongly suggest that these New World primates have developed a resistant innate immune response mechanism capable of controlling the macrophage intracellular growth of L. (L.) i. chagasi-infection, which do not encourage their use as animal model for studying AVL.

Susceptibility of peritoneal macrophage from different species of neotropical primates to Ex vivo Leishmania (L.) infantum chagasi-infection / Susceptibilidade do macrófago peritoneal de diferentes espécies de primatas neotropicais para a infecção ex vivo por Leishmania (L.) infantum chagasi

This study examined the susceptibility of peritoneal macrophage (PM) from the Neotropical primates: Callithrix jacchus, Callithrix penicillata, Saimiri sciureus, Aotus azarae infulatus and Callimico goeldii to ex vivo Leishmania (L.) infantum chagasi-infection, the etiological agent of American visceral leishmaniasis (AVL), as a screening assay for evaluating the potential of these non-human primates as experimental models for studying AVL. The PM-susceptibility to infection was accessed by the PM-infection index (PMI) at 24, 72 h and by the mean of these rates (FPMI), as well as by the TNF-α, IL-12 (Capture ELISA) and Nitric oxide (NO) responses (Griess method). At 24h, the PMI of A. azarae infulatus (128) was higher than those of C. penicillata (83), C. goeldii (78), S. sciureus (77) and C. jacchus (55). At 72h, there was a significant PMI decrease in four monkeys: A. azarae infulatus (128/37), C. penicillata (83/38), S. sciureus (77/38) and C. jacchus (55/12), with exception of C. goeldii (78/54). The FPMI of A. azarae infulatus (82.5) and C. goeldii (66) were higher than C. jacchus (33.5), but not higher than those of C. penicillata (60.5) and S. sciureus (57.5). The TNF-a response was more regular in those four primates which decreased their PMI at 24/72 h: C. jacchus (145/122 pg/mL), C. penicillata (154/130 pg/mL), S. sciureus (164/104 pg/mL) and A. azarae infulatus (154/104 pg/mL), with exception of C. goeldii (38/83 pg/mL). The IL-12 response was mainly prominent in A. infulatus and C. goeldii which presented the highest FPMI and, the NO response was higher in C. goeldii, mainly at 72 h. These findings strongly suggest that these New World primates have developed a resistant innate immune response mechanism capable of controlling the macrophage intracellular growth of L. (L.) i. chagasi-infection, which do not encourage their use as animal model for studying AVL.

Este estudo examinou a susceptibilidade do macrófago peritoneal (PM) dos primatas neotropicais: Callithrix jacchus, Callithrix penicillata, Saimiri sciureus, Aotus azarae infulatus e Callimico goeldii para a infecção ex vivo por Leishmania (L.) infantum chagasi, o agente etiológico da leishmaniose visceral americana (LVA), como método de triagem para avaliar o potencial desses primatas como modelo de estudo da LVA. A susceptibilidade do PM para a infecção foi investigada através do índice de infecção do PM (PMI) a intervalos de 24, 72 horas e, ainda, pela média dessas taxas (FPMI), assim como, pelas respostas do TNF-α, IL-2 (ELISA de captura) e óxido nítrico (NO) (método de Griess). Às 24hs da infecção experimental, o PMI do primata A. azarae infulatus (128) foi maior que aqueles de C. penicillata (83), C. goeldii (78), S. sciureus (77) e C. jacchus (55). Às 72hs, houve uma redução significativa do PMI de quatro primatas: A. azarae infulatus (128/37), C. penicillata (83/38), S. sciureus (77/38) e C. jacchus (55/12), com exceção de C. goeldii (78/54). O FPMI dos primatas A. azarae infulatus (82.5) e C. goeldii (66) foi maior que do primata C. jacchus (33.5), porém, não foi maior que dos primatas C. penicillata (60.5) e S. sciureus (57.5). A resposta do TNF-α foi mais regular nos quatro primatas que reduziram o PMI no intervalo de 24-72hs: C. jacchus (145/122 pg/µL), C. penicillata (154/130 pg/µL), S. sciureus (164/104 pg/µL) e A. azarae infulatus (154/104 pg/µL), com exceção de C. goeldii (38/83 pg/µL). A resposta de IL-12 foi, principalmente, marcante nos primatas A. azarae infulatus e C. goeldii, os quais apresentaram as maiores taxas do FPMI, e a resposta do NO foi maior no primata C. goeldii, em especial no intervalo de 72hs. Estes achados sugerem, fortemente, que estes primatas neotropicais parecem ter desenvolvido mecanismos resistentes de resposta imune inata capaz de controlar o crescimento intracelular da infecção por L. (L.) i. chagasi no macrófago, o que não encoraja o uso destes primatas como modelo de estudo da LVA.

Human cutaneous leishmaniasis: interferon-dependent expression of double-stranded RNA-dependent protein kinase (PKR) via TLR2.

We investigated the type I interferon (IFN-1)/PKR axis in the outcome of the Leishmania (Leishmania) amazonensis infection, along with the underlying mechanisms that trigger and sustain this signaling pathway. Reporter assays of cell extracts from RAW-264.7 macrophages infected with L. (L.) amazonensis or HEK-293T cells cotransfected with TLR2 and PKR promoter constructions were employed. Primary macrophages of TLR2-knockout (KO) or IFNR-KO mice were infected, and the levels of PKR, IFN-1, and superoxide dismutase 1 (SOD1) transcript levels were investigated and compared. Immunohistochemical analysis of human biopsy lesions was evaluated for IFN-1 and PKR-positive cells. Leishmania infection increased the expression of PKR and IFN-ß on induction of PKR-promoter activity. The observed effects required the engagement of TLR2. TLR2-KO macrophages expressed low IFN-ß and PKR levels postinfection with a reduced parasite load. We also revealed the requirement of PKR signaling for Leishmania-induced IFN-1 expression, responsible for sustaining PKR expression and enhancing infection. Moreover, during infection, SOD1 transcripts increased and were also enhanced when IFN-1 was added to the cultures. Remarkably, SOD1 expression was abrogated in infected, dominant-negative PKR-expressing cells. Finally, lesions of patients with anergic diffuse cutaneous leishmaniasis exhibited higher levels of PKR/IFN-1-expressing cells compared to those with single cutaneous leishmaniasis. In summary, we demonstrated the mechanisms and relevance of the IFN-1/PKR axis in the Leishmania infection.

Evaluation of Helicobacter pylory colonization by serologic test (IgG) and dyspepsia in volunteers from the countryside of Monte Negro, in the Brazilian western Amazon region / Avaliação da colonização por Helicobacter pylori através de teste sorológico (IgG) e de dispepsia em voluntários da população rural de Monte Negro (RO), região da Amazônia ocidental

The present study intended to analyze the seroprevalence of Helicobacter pylori, IgG, and its relation to dyspepsia in a population from the western Amazon region. During the "Projeto Bandeira Científica", a University of São Paulo Medical School program, in Monte Negro's rural areas, state of Rondônia, 266 blood samples were collected from volunteers. The material was tested for IgG antibodies anti-Helicobacter pylori by ELISA method and the participants were also interviewed on dyspepsia, hygiene and social aspects. Participants aged between five and 81 years old (34 years on average), 149 (56 percent) were female and 117 (44 percent) male. We found 210 (78.9 percent) positive, 50 (18.8 percent) negative and six (2.3 percent) undetermined samples. Dyspeptic complaints were found in 226 cases (85.2 percent). There was no statistical association between dyspepsia and positive serology for H. pylori. We concluded that the seroprevalence in all age categories is similar to results found in other studies conducted in developing countries, including those from Brazil. On the other hand, the seroprevalence found in Monte Negro was higher than that reported in developed countries. As expected, there was a progressive increase in the positivity for H. pylori in older age groups.

Este trabalho tem por objetivo analisar a soroprevalência do Helicobacter pylori, IgG, em população rural da Amazônia, e sua correlação com queixa dispéptica. No Projeto Bandeira Científica da FMUSP, em Monte Negro - RO, foram coletadas 266 amostras sangüíneas nos assentamentos rurais do município. Foram pesquisados anticorpos da classe IgG dirigidos contra Helicobacter pylori pelo método ELISA e aplicados questionários sobre dispepsia, aspectos sociais e epidemiológicos. Os pacientes tinham idades entre cinco e 81 anos (média de 34 anos); 149 (56 por cento) do sexo feminino e 117 (44 por cento) do sexo masculino. Foram encontradas 210 (78.9 por cento) amostras positivas, 50 negativas (18.8 por cento) e seis indeterminadas (2.3 por cento). A queixa de dispepsia foi encontrada em 226 casos (85.2 por cento). Não houve associação significativa entre os sintomas dispépticos e a soro positividade para H. pylori. Concluímos que a soro prevalência para todas as faixas etárias é comparável com os resultados de outros estudos realizados em países em desenvolvimento, e maior que aquela encontrada nos países desenvolvidos. Houve aumento progressivo da positividade com a idade, como citado na literatura.

Endemic hepatitis b and c virus infection in a brazilian eastern amazon region

CONTEXT: Hepatitis B and hepatitis C infection has been an important cause of morbidity and mortality around the world. However there are few investigations regarding the prevalence and possible risk factors for these diseases in Brazil, particularly in Amazon region, where there are some endemic focus. OBJECTIVES: To determine the prevalence of hepatitis B and hepatitis C in the city of Buriticupu, MA, located in the Brazilian Eastern Amazon region, and try to explore the risk factors for these infections in that area. METHODS: Two hundred forty three subjects (46.5 percent male and 53.5 percent female) were investigated. RESULTS: The prevalence of past or current infection of hepatitis B and C virus was, respectively, 40.74 percent and 5.76 percent. Positivity for HBsAg was found in 2.88 percent of the subjects. The prevalence of current infection or chronic virus carriers found was 2.88 percent (HBsAg). There was a statistically significant relationship between the sera-prevalence of anti-HBc and the distance of the residence from the city center which may reflect an indirect association between the infection and precarious conditions of existence. Individuals with age equal or greater than 60 years were also more likely to be anti-HBc positive which could only reflect that older people have a longer history of exposure to hepatitis B infection. The prevalence of hepatitis C is higher than the worldwide estimate. CONCLUSION: Buriticupu may be considered endemic for hepatitis B and C. Hepatitis B infection could be related to precarious living conditions and old age. Hepatitis C was not associated with the variables investigated in the present investigation.

CONTEXTO: Infecção por hepatites B e C tem sido causa importante de morbimortalidade em todo o mundo. Entretanto, há poucas investigações sobre a prevalência e possíveis fatores de risco relacionados a tais doenças no Brasil, especialmente na região amazônica, onde há algumas regiões endêmicas para tais quadros clínicos. OBJETIVOS: Detectar a prevalência de hepatites B e C na cidade de Buriticupu, MA, localizada na região leste da Amazônia brasileira, e tentar investigar seus fatores de risco nessa área. MÉTODOS: Duzentos e quarenta e três indivíduos (46,5 por cento masculinos e 53,5 por cento femininos) foram investigados. RESULTADOS: A prevalência de hepatite C foi de 5,71 por cento (anti-HCV) e a de hepatite B foi de 40,74 por cento (anti-HBc). A prevalência de indivíduos com infecção atual ou com infecção crônica foi de 2,80 por cento (HBsAg). Houve relação estatisticamente significante entre anti-HBc e a distância da residência dos indivíduos do centro da cidade, o que pode refletir uma associação indireta entre tal quadro infeccioso e condições precárias de existência. Indivíduos com idade igual ou maior a 60 anos também apresentaram maior chance de apresentarem sorologia para anti-HBc, o que pode refletir apenas que pessoas mais velhas apresentam história maior de exposição à infecção. A prevalência de hepatite C é maior do que a mundial estimada. CONCLUSÃO: Buriticupu pode ser considerada endêmica para hepatites B e C. Hepatite B pode estar relacionada com precárias condições de vida e idade avançada. Hepatite C não foi associada com as variáveis investigadas na presente investigação.

Histopathology, humoral and cellular immune response in the murine model of Leishmania (Viannia) shawi.

Leishmania (Viannia) shawi was recently characterized and few studies concerning modifications in cellular and humoral immune responses in experimental leishmaniasis have been conducted. In this work, immunopathological changes induced by L. shawi in chronically infected BALB/c mice were investigated. Infected BALB/c mice developed increased lesion size associated with strong inflammatory infiltrate diffusely distributed in the dermis, with highly infected macrophages. The humoral immune response was predominantly directed toward the IgG1 isotype. The functional activity of CD4(+) and CD8(+) T cells showed significantly increased TNF-alpha mRNA levels associated with reduced IFN-gamma expression by CD4(+) T cells and the double negative (dn) CD4CD8 cell subset. High IL-4 levels expressed by CD8(+) T cells and dnCD4CD8 and TGF-beta by CD4(+) and CD8(+) T cells were detected, while IL-10 was highly expressed by all three cell subpopulations. Taken together, these results show an evident imbalance between TNF-alpha and IFN-gamma that is unfavorable to amastigote replication control. Furthermore, L. shawi seems to regulate different cell populations to express deactivating cytokines to avoid its own destruction. This study indicates BALB/c mice as a potentially good experimental model for further studies on American cutaneous leishmaniosis caused by L. shawi.

Leishmania chagasi Cunha & Chagas, 1937: indigenous or introduced? A brief review

This review focused on the etiology of American visceral leishmaniasis due to a recent polemic regarding the origin of its etiological agent, Leishmania chagasi Cunha & Chagas, 1937. This parasite was described as a new Leishmania species in light of its inability to produce experimentally visceral disease in domestic dogs; this characteristic distinguished it from the other prior known etiologic agent of visceral leishmaniasis in the Mediterranean Basin of Europe, Leishmania infantum Nicolle, 1908. After 50 years of Leishmania chagasi investigation, the genus Leishmania was reviewed and the parasite was reclassified as a member of the subgenus Leishmania, species Leishmania (Leishmania) chagasi. Recently, after molecular analysis using the random amplified polymorphic DNA (RAPD) technique that compared L. (L.) chagasi with L. (L.) infantum, it was concluded that these species were genetically indistinguishable and, therefore, L. (L.) chagasi was regarded as synonymous with L. (L.) infantum. For this reason, this review has evaluated all knowledge concerning the eco-epidemiology of L. (L.) chagasi in the Brazilian Amazon, principally in regard to the sylvatic habits of its phlebotomine sandfly vector, Lutzomyia longipalpis, and its vertebrate reservoir, the wild fox Cerdocyon thous, with the aim of showing that L. (L.) chagasi cannot be neglected from the parasitological investigation of visceral leishmaniasis in the New World; it must be considered, at least, at the subspecific level as Leishmania (L.) infantum chagasi...

Esta revisão aborda a etiologia da leishmaniose visceral americana devido a uma recente polêmica sobre a origem do seu agente etiológico, a Leishmania chagasi Cunha & Chagas, 1937. Conforme é sabido, este parasito foi descrito como uma nova espécie de Leishmania em razão da sua incapacidade de produzir, experimentalmente, a leishmaniose visceral no cão doméstico; este caráter a diferenciou de outro agente etiológico já conhecido da leishmaniose visceral na Bacia do Mediterrâneo na Europa: Leishmania infantum Nicolle, 1908. Após 50 anos da descrição da Leishmania chagasi, o gênero Leishmania sofreu ampla revisão e o parasito foi reclassificado como um membro do subgênero Leishmania, espécie Leishmania (Leishmania) chagasi. Recentemente, em seguida a uma análise molecular usando a técnica da amplificação aleatória polimórfica do DNA (RAPD), que comparou a L. (L.) chagasi com a L. (L.) infantum, foi concluído que ambos os parasitos eram geneticamente indistinguíveis e, portanto, que a L. (L.) chagasi era sinônimo de L. (L.) infantum. Por esse motivo, esta revisão procurou agregar todo o conhecimento sobre a eco-epidemiologia da L. (L.) chagasi na Amazônia brasileira, principalmente acerca dos hábitos silvestres de seu flebotomíneo vetor, Lutzomyia longipalis, e seu reservatório vertebrado, a raposa do campo Cerdocyon thous, com o propósito de demonstrar que a L. (L.) chagasi não pode ser negligenciada do cenário parasitológico da leishmaniose visceral no Novo Mundo; ela deve ser considerada, pelo menos em um nível subespecífico, como Leishmania (L.) infantum chagasi...

American cutaneous leishmaniasis in the Pontal of Paranapanema - SP, Brazil: ecological and entomological aspects / Leishmaniose cutânea americana no Pontal do Paranapanema, SP, Brasil: aspectos ecológicos e entomológicos

American cutaneous leishmaniasis (ACL) occurs in epidemic outbreaks and in sporadic cases with small annual variation in the Pontal of Paranapanema, SP. There is little research on the sandfly fauna of this region. The last outbreaks were related to the Movement of the Landless Workers (MST) and with the ecological tourism in preserved forest of the Parque Estadual do Morro do Diabo (PEMD). AIM: identification of the sandfly fauna within the PEMD, mainly anthropophilic species already incriminated as vectors of ACL, as well as their seasonality, hourly frequency and data of the behavior. M&M: The captures were undertaken with CDC light and Shannon traps from 6:00 pm to 10:00 pm, monthly from May 2000 to December 2001. The temperature and relative humidity data were registered at hourly intervals. RESULTS: The captured species were: Brumptomyia brumpti, Nyssomyia neivai, Nyssomyia whitmani, Pintomyia fischeri and Pintomyia pessoai. The P. pessoai predominated (34.39 percent) and N. neivai was less found (0.74 percent), only being captured in CDC traps. Shannon trap captured more sandflies (63.01 percent) than the CDC traps (36.99 percent). Despite the environmental degradation anthropophilic species, indicates favorable bioecological conditions for persistence of vectors and potential transmission of leishmaniasis.

LCA ocorre em surtos epidêmicos e casos esporádicos com pequena variação anual no Pontal do Paranapanema, SP. Há pouca pesquisa sobre a fauna flebotomínea na região. Os últimos surtos estão relacionados ao MST e turismo ecológico na floresta do Parque Estadual do Morro do Diabo (PEMD). OBJETIVO: identificar a fauna flebotomínea no PEMD, principalmente as espécies antropofílicas já incriminadas como vetores da LCA, também, sua sazonalidade, horários e dados de comportamento. M&M: As capturas foram feitas com armadilhas CDC e Shannon das 18 às 22h, mensalmente de maio de 2000 a dezembro de 2001. Dados de temperatura e umidade relativa foram registrados com intervalo de uma hora. RESULTADOS: As espécies capturadas foram: Brumptomyia brumpti, Nyssomyia neivai, Nyssomyia whitmani, Pintomyia fischeri e Pintomyia pessoai. A P. pessoai predominou (34,39 por cento) e a N. neivai foi a menos encontrada (0,74 por cento), capturada com a CDC. A Shannon capturou mais flebotomíneos (63,01 por cento) que a CDC (36,99 por cento). Apesar da degradação ambiental espécies, antropofílicas foram encontradas, indicando condições bioecológicas para persistência de vetores e potencial transmissão da leishmaniose.

Saliva of laboratory-reared Lutzomyia longipalpis exacerbates Leishmania (Leishmania) amazonensis infection more potently than saliva of wild-caught Lutzomyia longipalpis.

In order to compare the saliva effect from wild-caught and lab-reared L. longipalpis on the development of experimental cutaneous leishmaniasis, C57BL/6 mice were inoculated subcutaneously into the hind footpads with promastigotes of L. (L.) amazonensis plus salivary gland lysate from wild-caught (SGL-W) and lab-colonized (SGL-C) vectors. Lesion sizes were significantly larger in the mice infected with both saliva compared to mice infected with parasites alone; moreover, the lesions caused by parasite+SGL-C were significantly larger than the lesions caused by parasite+SGL-W. Histopathological morphometric studies regarding the acute phase of infections showed lower numbers of polymorphonuclear cells, greater numbers of mononuclear cells and parasites in SGL-C infected mice compared to SGL-W infected mice. In the chronic phase of infection, the number of mononuclear cells was lower and the number of parasites was greater in SGL-C infected mice than SGL-W infected mice. In vitro studies showed increased infection index of macrophages infected with parasites plus saliva compared to infection with parasites alone, with no difference between the saliva infection indices. SDS-PAGE gel for SGL-C and SGL-W showed differences in the composition and quantity of protein bands, determined by densitometry. These results call attention to the experimental saliva model, which shows exacerbation of infection caused by sandfly saliva.

Mucin pattern reflects the origin of the adenocarcinoma in Barrett's esophagus: a retrospective clinical and laboratorial study.

BACKGROUND: Mucin immunoexpression in adenocarcinoma arising in Barrett's esophagus (BE) may indicate the carcinogenesis pathway. The aim of this study was to evaluate resected specimens of adenocarcinoma in BE for the pattern of mucins and to correlate to the histologic classification. METHODS: Specimens were retrospectively collected from thirteen patients who underwent esophageal resection due to adenocarcinoma in BE. Sections were scored for the grade of intestinal metaplasia. The tissues were examined by immunohistochemistry for MUC2 and MUC5AC antibodies. RESULTS: Eleven patients were men. The mean age was 61 years old (varied from 40 to 75 years old). The tumor size had a mean of 4.7 +/- 2.3 cm, and the extension of BE had a mean of 7.7 +/- 1.5 cm. Specialized epithelium with intestinal metaplasia was present in all adjacent mucosas. Immunohistochemistry for MUC2 showed immunoreactivity in goblet cells, while MUC5AC was extensively expressed in the columnar gastric cells, localizing to the surface epithelium and extending to a variable degree into the glandular structures in BE. Tumors were classified according to the mucins in gastric type in 7/13 (MUC5AC positive) and intestinal type in 4/13 (MUC2 positive). Two tumors did not express MUC2 or MUC5AC proteins. The pattern of mucin predominantly expressed in the adjacent epithelium was associated to the mucin expression profile in the tumors, p = 0.047. CONCLUSION: Barrett's esophagus adenocarcinoma shows either gastric or intestinal type pattern of mucin expression. The two types of tumors developed in Barrett's esophagus may reflect the original cell type involved in the malignant transformation.

Prevalência de lombalgia em garimpeiros de Serra Pelada, Pará / Brasil / Prevalence of low back pain among gold diggers in Serra Pelada, Pará / Brazil

O presente estudo avalia a prevalência de lombalgia em população de garimpeiros de Serra Pelada, localizada no Estado do Pará / Brasil. Considerado como um dos maiores garimpos de ouro na década de 80, havendo em torno de 100.000 pessoas envolvidas na extração do ouro. Atualmente a população local é estimada em 6000 habitantes. Através de uma avaliação médica primária populacional feita em 2002 foi realizado um estudo de corte transversal retrospectivo com a análise de dados obtidos durante esse atendimento. Foram 1.169 atendimentos, 605 do sexo masculino (51,7 por cento), idade média de 41,4 ± 21,2 anos. 47 por cento responderam que trabalham ou trabalharam no garimpo, com tempo médio de 10,9 ± 9,7 anos. A prevalência de lombalgia foi de 13,7 por cento em relação ao geral dos diagnósticos. Quando comparada ao grupo de garimpeiros, a prevalência encontrada foi de 18,2 por cento contra 10,0por cento em relação ao grupo de não garimpeiros (p<0,000), porém não mantendo significância estatística após a regressão logística. Estudos mais detalhados seriam necessários para tentar estabelecer as possíveis relações com dor lombar nesta população, sendo que talvez o rastreamento de antigos garimpeiros pudesse investigar melhor uma possível relação direta laboral do garimpo com lombalgia.

Macrophage and TGF-beta immunohistochemical expression in Jorge Lobo's disease.

Jorge Lobo's disease, or lacaziosis, is a chronic deep mycosis that clinically manifests as solid, variable-sized nodular parakeloidal lesions. Few studies have characterized the in situ cellular and humoral immune response, especially the involvement of cytokines with immunosuppressive effects such as TGF-beta. The objective this paper was to analyze the expression of TGF-beta in cutaneous lesions in lacaziosis and investigate its importance in the etiopathogy of the disease. The results indicate that the abundance of collagen bands, together with weak immunolabeling for CD68 seen in macrophages, indicates a concomitant effect of TGF-beta inhibiting macrophages and inducing fibrosis, which is responsible for the keloid aspect frequently acquired by these lesions. Finally, the evolution of the infection supports the hypothesis that TGF-beta plays a fundamental role in the etiopathology of Lacazia loboi infection, either by inhibiting the cellular immune response mainly mediated by macrophages or by inducing fibrosis. Further studies are necessary to better characterize the phenotype of the inflammatory infiltrate as well as the participation of other cytokines and growth factors in the tissue response of the host in Jorge Lobo's disease.

Health evaluation of gold miners living in a mercury-contaminated village in Serra Pelada, Pará, Brazil.

Serra Pelada is a village in the Amazon region of Brazil where most of the inhabitants are former gold miners. Of 235 individuals evaluated, 219 were males (93.19%), 16 were females (6.80%), and the mean age was 52.07 years (standard deviation = 11.57). Most were heavy drinkers (62.44%) and smokers (70.30%), and 85.53% had previously suffered from malaria. Reported symptoms included fatigue (30.60%), irritability (35.62%), excitability (14.16%), insomnia (34.48%), memory loss (61.80%), visual field constriction (4.18%), paresthesia (64.93%), partial hearing loss (16.35%), and gingivitis (18.01%). After an examination of the residents, the authors observed several neurological symptoms: tremors (22.80%), involuntary ocular movement (2.20%), visual field constriction (4.18%), Romberg syndrome (2.33%), involuntary tongue movement (2.19%), dysdiadochokinesia (0.43%), failure of a finger-nose test (10.96%), failure of a knee-heel test (4.84%), inability to complete a tandem march (6.25%), muscular weakness (2.27%), and damage to sensory organs (24.66%). The authors concluded that these neurological changes possibly resulted from mercury toxicity; however, they could not determine a significant correlation with the mercury levels detected in participants' urine.

hMLH1, hMSH2 and cyclooxygenase-2 (cox-2) in sporadic colorectal polyps.

BACKGROUND: Colorectal adenomatous polyps are known as premalignant lesions. Mutations in the mismatch repair (MMR) enzymes hMLH1, hMSH2 and hMSH6 are recognized causes of hereditary non-polyposis colorectal cancer and act by inducing a mutator phenotype characterized by microsatellite instability (MSI). MSI is also detected in sporadic colorectal cancers. Cox-2 is an inducible enzyme that regulates prostaglandin synthesis and it is overexpressed at sites of inflammation, in colorectal adenomatous polyps and cancer. The aim of this study was to evaluate the immunoexpression of hMLH1, hMSH2 and Cox-2 in polyps resected through colonoscopy, and to examine their association with clinicopathological characteristics (age, gender, location, size, histology and grade of dysplasia). PATIENTS AND METHODS: One hundred and sixty-seven colonic polyps, 6 normal colonic mucosa samples, and 23 samples of colorectal adenocarcinoma were used in this study. All patients had no family history of colorectal cancer. The samples were prospectively collected and immunostained for hMLH1, hMSH2 and Cox-2 using the ABC-immunohistochemistry technique with amplification by biotinylated tyramide. The mean age was 60.2+/-13.8 years (range 21-90 years) and 77 (55.8%) were men. RESULTS: Tubular adenomas were present in 81.4%, tubulous-villous in 15.9%, serrated in 1.8%, and villous in 0.9%. The majority of the adenomas were located in the rectosigmoid region (63.5%), followed by ascendant in 14.2%, cecum in 7.5%, descendent in 8.2% and transverse in 6.7%. Low-grade dysplasia was detected in 59.6% of the adenomas. Loss of hMLH1 and hMLH2 immunoexpression was observed in 20% and 15.5% of the adenomas, respectively. Cox-2 expression was found in 9% of the adenomas, and in 40% of the adenocarcinomas. Moreover, Cox-2 immunoexpression was associated with the multiplicity of adenomas in the same patient (p=0.001). There was no association between marker immunoexpression and gender, age, location, size, histology or grade of dysplasia. CONCLUSION: Loss of hMLH1 and hMLH2 immunoexpression in adenomas is relatively frequent in patients without colorectal cancer family history. Cox-2 is overexpressed in colorectal adenomatous polyps and adenocarcinomas, and its positivity in adenomas may indicate a higher risk for multiple lesions.

Microsporidiosis in a Brazilian University Hospital: case report

This is the report on a patient with chronic diarrhea caused by microsporidia. He is married, infected with HIV and has low CD4 cell count. The diagnosis was established through stool parasite search using concentration methods and Gram - chromotrope staining technique. Ileum biopsy was also performed in this case. The etiological diagnosis may be established in a clinical laboratory, by chromotrope staining technique in routine microscopic examination of stool specimens.

Este é o relato de caso de doente com diarréia crônica causada por Microsporidia. O doente era homem, casado, infectado com HIV e tinha baixa taxa de linfócitos CD4+. O diagnóstico foi feito em exame de fezes utilizando métodos de concentração e técnica de coloração de Gram-Chromotrope. Biópsia de íleo também foi realizada neste caso. O diagnóstico etiológico pode ser feito em laboratório clínico, por técnicas de coloração baseada em cromotrope na rotina da observação microscópica direta.

Multilocus genotyping of Cryptosporidium hominis associated with diarrhea outbreak in a day care unit in São Paulo

Mundialmente, diferentes espécies de Cryptosporidium estão relacionadas com doenças diarréicas. No Brasil há poucos dados sobre os genótipos das espécies de Cryptosporidium associadas a infecções. OBJETIVO: No presente estudo, caracterizamos, por métodos moleculares, a espécie e o genótipo de Cryptosporidium sp diagnosticado em surto diarréico ocorrido na creche do Hospital das Clínicas, São Paulo, Brasil. MATERIAL E MÉTODOS: Identificação específica e tipagem dos isolados associados ao surto foram feitos a partir do seqüenciamento de fragmentos de DNA amplificados por PCR dos seguintes loci: a região que codifica o SSUrRNA, o gene que codifica uma proteína do envoltório dos oocistos de Cryptosporidium (COWP), e o locus de microsatélite ML1, representado por seqüências repetitiva de três nucleotídeos GAG contendo substituições que diferem entre os genótipos de Cryptosporidium parvum e Cryptosporidium hominis. RESULTADOS: Um total de 29 amostras positivas para Cryptosporidium associadas ao surto diarréico foi analisado com base nos métodos moleculares acima descritos. O estudo revelou a presença do genótipo ML1 de Cryptosporidium hominis. DISCUSSÃO: A análise molecular reforçou a hipótese de que a transmissão de Cryptosporidium hominis durante o surto diarréico ocorreu de pessoa a pessoa através da rota fecal oral. Esta é a primeira vez que ferramentas moleculares são utilizadas para identificação de espécies e genótipos de isolados acusando a presença do genótipo ML1 em pacientes brasileiros.