RESUMO
In Central America, infection by Leishmania (Leishmania) infantum chagasi causes visceral leishmaniasis and non-ulcerated cutaneous leishmaniasis (NUCL). This work aimed to evaluate the participation of subpopulations of antigen-presenting cells in skin lesions of patients affected by NUCL through double-staining immunohistochemistry using cellular and intracellular markers. Twenty-three skin biopsies from patients affected by NUCL were used. Histological sections stained by HE were used for histopathological study. Immunohistochemical studies were performed using primary antibodies against Langerhans cells, dermal dendritic cells, T lymphocytes, and the cytokines IL-12, IFN-γ, TNF-α, iNOS, and IL-10. The histopathological lesions were characterized by an inflammatory infiltrate, predominantly lymphohistiocytic, of variable intensity, with a diffuse arrangement associated with epithelioid granulomas and discreet parasitism. Double-staining immunohistochemistry showed higher participation of dendritic cells producing the proinflammatory cytokine IL-12 in relation to the other evaluated cytokines. Activation of the cellular immune response was marked by a higher density of CD8 Tc1-lymphocytes followed by CD4 Th1-lymphocytes producing mainly IFN-γ. The data obtained in the present study suggest that antigen-presenting cells play an important role in the in situ immune response through the production of proinflammatory cytokines, directing the cellular immune response preferentially to the Th1 and Tc1 types in NUCL caused by L. (L.) infantum chagasi.
Assuntos
Leishmania infantum , Leishmaniose Cutânea , Leishmaniose Visceral , Humanos , Citocinas , Células Apresentadoras de Antígenos , Interleucina-12RESUMO
Macrophages play important roles in the innate and acquired immune responses against Leishmania parasites. Depending on the subset and activation status, macrophages may eliminate intracellular parasites; however, these host cells also can offer a safe environment for Leishmania replication. In this sense, the fate of the parasite may be influenced by the phenotype of the infected macrophage, linked to the subtype of classically activated (M1) or alternatively activated (M2) macrophages. In the present study, M1 and M2 macrophage subsets were analyzed by double-staining immunohistochemistry in skin biopsies from patients with American cutaneous leishmaniasis (ACL) caused by L. (L.) amazonensis, L. (V.) braziliensis, L. (V.) panamensis ,and L. (L.) infantum chagasi. High number of M1 macrophages was detected in nonulcerated cutaneous leishmaniasis (NUCL) caused by L. (L.) infantum chagasi (M1 = 112 ± 12, M2 = 43 ± 12 cells/mm2). On the other side, high density of M2 macrophages was observed in the skin lesions of patients with anergic diffuse cutaneous leishmaniasis (ADCL) (M1 = 195 ± 25, M2 = 616 ± 114), followed by cases of localized cutaneous leishmaniasis (LCL) caused by L. (L.) amazonensis (M1 = 97 ± 24, M2 = 219 ± 29), L. (V.) panamensis (M1 = 71 ± 14, M2 = 164 ± 14), and L. (V.) braziliensis (M1 = 50 ± 13, M2 = 53 ± 10); however, low density of M2 macrophages was observed in NUCL. The data presented herein show the polarization of macrophages in skin lesions caused by different Leishmania species that may be related with the outcome of the disease.
Assuntos
Leishmania/imunologia , Leishmaniose Cutânea/imunologia , Ativação de Macrófagos , Macrófagos/imunologia , Pele/parasitologia , Biópsia , Humanos , Leishmaniose Cutânea/parasitologia , Leishmaniose Cutânea/patologia , Macrófagos/parasitologia , Pele/imunologia , Pele/patologiaRESUMO
The induced expression of nitric oxide synthase (iNOS) controls the intracellular growth of Leishmania in infected macrophages. Histones deacetylases (HDACs) negatively regulate gene expression through the formation of complexes containing transcription factors such as NF-κB p50/50. Herein, we demonstrated the occupancy of p50/p50_HDAC1 to iNOS promoter associated with reduced levels of H3K9Ac. Remarkably, we found increased levels of HDAC1 in L. amazonensis-infected macrophages. HDAC1 upregulation was not found in L. major-infected macrophages. The parasite intracellular load was reduced in HDAC1 knocked-down macrophages, which presented increased nitric oxide levels. HDAC1 silencing led to the occupancy of CBP/p300 to iNOS promoter and the rise of H3K9Ac modification. Importantly, the immunostaining of skin samples from hiporeactive cutaneous leishmaniasis patients infected with L. amazonensis, revealed high levels of HDAC1. In brief, L. amazonensis induces HDAC1 in infected macrophages, which contribute to parasite survival and is associated to hiporeactive stage found in L. amazonensis infected patients.
Assuntos
Histona Desacetilase 1/metabolismo , Leishmania braziliensis/fisiologia , Leishmaniose Cutânea/imunologia , Macrófagos/imunologia , Óxido Nítrico Sintase Tipo II/metabolismo , Pele/patologia , Adolescente , Adulto , Células Cultivadas , Criança , Extinção Biológica , Feminino , Histona Desacetilase 1/genética , Interações Hospedeiro-Parasita , Humanos , Evasão da Resposta Imune , Leishmaniose Cutânea/genética , Masculino , Pessoa de Meia-Idade , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Carga Parasitária , Regiões Promotoras Genéticas/genética , Ligação Proteica , RNA Interferente Pequeno/genética , Fator de Transcrição Sp1/metabolismo , Adulto JovemRESUMO
BACKGROUND: Mucosal alterations after Roux-en-Y gastric bypass for morbid obesity have not been clearly evaluated. This study aims to analyze the mucosal alterations (proliferative status (Ki-67); apoptosis (caspase-3 and BCL-2); hormonal function (gastrin)) in the excluded stomach. METHODS: Double-balloon enteroscopy was performed in 35 patients who underwent Roux-en-Y gastric bypass longer than 36 months. Multiple biopsies of the proximal pouch and the excluded gastric mucosa were collected. Gastric biopsies from 32 non-operated obese patients were utilized as controls. Endoscopic biopsies were cut from tissue blocks fixed in formalin and embedded in paraffin. Sections 4 µm thick were examined for immunoexpression using the streptavidin-biotin-peroxidase method. RESULTS: The two groups were comparable for age, gender, gastritis, intestinal metaplasia, and Helicobacter pylori. The mean number of positive gastrin cells was 55.5 (standard deviation (SD) = 11.7) in the control group and 29.6 (SD = 7.9) in the cases, p = 0.0003. Ki-67 proliferative index in cases (body = 24.7%, antrum = 24.9%) was significantly higher compared to controls (body = 15.0% and antrum = 17.7%), p = 0.002 and 0.01, respectively. Caspase-3 immunoexpression was higher in the controls compared to the excluded stomach (46 vs. 31%), p = 0.02. There was no statistical difference between CD3, CD8, and Bcl-2 immunoexpressions in the control and cases. CONCLUSIONS: Cell proliferation is increased and apoptosis is downregulated in the excluded gastric mucosa compared to the non-operated obese controls. Alterations in cell turnover and in hormonal secretions in these conditions may be of relevance in long-term follow-up.
Assuntos
Células Epiteliais/patologia , Derivação Gástrica , Mucosa Gástrica/patologia , Coto Gástrico/patologia , Obesidade Mórbida/patologia , Adolescente , Adulto , Idoso , Brasil , Caspase 3/metabolismo , Proliferação de Células , Enteroscopia de Duplo Balão , Regulação para Baixo , Feminino , Mucosa Gástrica/microbiologia , Gastrinas/metabolismo , Gastrite/patologia , Infecções por Helicobacter/patologia , Helicobacter pylori/isolamento & purificação , Humanos , Imuno-Histoquímica , Antígeno Ki-67/metabolismo , Masculino , Pessoa de Meia-Idade , Obesidade Mórbida/microbiologia , Obesidade Mórbida/cirurgiaRESUMO
In Amazonian Brazil, the Cebus apella monkey (Primates: Cebidae) has been associated with the enzootic cycle of Leishmania (V.) shawi, a dermotropic parasite causing American cutaneous leishmaniasis (ACL). It has also been successfully used as animal model for studying cutaneous leishmaniasis. In this work, there has been investigated its susceptibility to experimental Leishmania (L.) infantum chagasi-infection, the etiologic agent of American visceral leishmaniasis (AVL). There were used ten C. apella specimens, eight adult and two young, four males and six females, all born and raised in captivity. Two experimental infection protocols were performed: i) six monkeys were inoculated, intra-dermal via (ID), into the base of the tail with 2 x 10(6) promastigotes forms from the stationary phase culture medium; ii) other four monkeys were inoculated with 3 x 10(7) amastigotes forms from the visceral infection of infected hamsters by two different via: a) two by intravenous via (IV) and, b) other two by intra-peritoneal via (IP). The parameters of infection evaluation included: a) clinical: physical exam of abdomen, weigh and body temperature; b) parasitological: needle aspiration of the bone-marrow for searching of amastigotes (Giemsa-stained smears) and promastigotes forms (culture medium); c) immunological: Indirect fluorescence antibody test (IFAT) and, Delayed-type hypersensitivity (DTH). In the six monkeys ID inoculated (promastigotes forms) all parameters of infection evaluation were negative during the 12 months period of follow-up. Among the four monkeys inoculated with amastigotes forms, two IV inoculated showed the parasite in the bone-marrow from the first toward to the sixth month p.i. and following that they cleared the infection, whereas the other two IP inoculated were totally negative. These four monkeys showed specific IgG-antibody response since the third month p.i. (IP: 1/80 and IV: 1/320 IgG) toward to the 12th month (IP: 1/160 and IV: 1/5120). The DTH-conversion occurred in only one IV inoculated monkey with a strong (30 mm) skin reaction. Considering these results, we do not encourage the use of C. apella monkey as animal model for studying the AVL.
Na Amazônia Brasileira, o macaco Cebus apella (Primata: Cebidae) tem sido associado com o ciclo enzoótico da Leishmania (V.) shawi, um parasito dermotrópico causador da Leishmaniose Tegumentar Americana (LTA). Ele tem sido também empregado com sucesso como modelo experimental para estudo da leishmaniose tegumentar. Neste trabalho, foi investigada sua susceptibilidade à infecção experimental por Leishmania (L.) infantum chagasi, o agente etiológico da Leishmaniose Visceral Americana (LVA). Foram usados dez espécimes de C. apella oito adultos e dois jovens, quatro machos e seis fêmeas, todos nascidos e criados em cativeiro. Dois protocolos de infecção experimental foram feitos: i) seis macacos foram inoculados por via intradérmica (ID), na base da cauda com 2x10(6) formas promastigotas em fase estacionária de crescimento; ii) outros quatro macacos foram inoculados com 3x10(7) formas amastigotas de infecção visceral de hamsteres por duas vias diferentes: a) dois por via intravenosa (IV) e, b) outros dois pela via intraperitoneal (IP). A avaliação da infecção incluiu parâmetros: clínico: exame físico do abdômen, peso e temperatura corporal; b) parasitológico: aspirado de medula óssea por agulha para procura de amastigotas (esfregaço corado por Giemsa) e formas promastigotas (meio de cultura); c) imunológico: Reação de Imunofluorescência Indireta (RIFI) e, resposta de hipersensibilidade tardia (DTH). Nos seis macacos inoculados ID (formas promastigotas) todos os parâmetros de avaliação da infecção foram negativos durante o período de 12 meses. Entre os quatro macacos inoculados com formas amastigotas, dois inoculados IV mostraram parasitos na medula óssea do primeiro ao sexto mês p.i. e em seguida houve a resolução da infecção, no entanto os outros dois inoculados IP foram totalmente negativos. Esses quatro macacos apresentaram resposta específica de anticorpo IgG desde o terceiro mês p.i. (IP: 1/80 e IV: 1/320) até o décimo segundo mês (IP: 1/160 e IV: 1/5120). A conversão DTH ocorreu em apenas um macaco inoculado IV com uma forte reação na pele (30 mm). Considerando esses resultados, nós não recomendamos o uso do macaco C. apella como modelo animal para estudo da LVA.
Assuntos
Animais , Feminino , Masculino , Anticorpos Antiprotozoários/imunologia , Cebus/parasitologia , Imunoglobulina G/imunologia , Leishmania infantum/patogenicidade , Leishmaniose Visceral/parasitologia , Doenças dos Macacos/parasitologia , Cebus/imunologia , Modelos Animais de Doenças , Suscetibilidade a Doenças , Técnica Indireta de Fluorescência para Anticorpo , Leishmaniose Visceral/imunologia , Doenças dos Macacos/imunologiaRESUMO
Canine visceral leishmaniasis (CVL) is recognizable by characteristic signs of disease and is highly lethal. The infection, however, may be quite inapparent in some seropositive dogs, and this has raised the polemic question as to whether or not such animals can be a source of infection for Lutzomyia longipalpis, the vector of American visceral leishmaniasis (AVL). In this study we have examined 51 dogs with acute CVL from an AVL area in Pará State, northern Brazil, and compared the parasite density, amastigotes of Leishmania (L.) infantum chagasi, in the skin, lymph node and viscera of symptomatic with that of nine asymptomatic but seropositive dogs (IFAT-IgG). Post-mortem biopsy fragments of these tissues were processed by immunohistochemistry, using a polyclonal antibody against Leishmania sp. The X² and Mann Whitney tests were used to evaluate the means of infected macrophage density (p < 0.05). There was no difference (p > 0.05) in the skin (10.7/mm² x 15.5/mm²) and lymph node (6.3/mm² x 8.3/mm²), between asymptomatic and symptomatic dogs, respectively. It was higher (p < 0.05), however, in the viscera of symptomatic (5.3/mm²) than it was in asymptomatic (1.4/mm²) dogs. These results strongly suggest that asymptomatic or symptomatic L. (L.) i. chagasi-infected dogs can serve as a source of infection, principally considering the highest (p < 0.05) parasite density from skin (10.7/mm² x 15.5/mm²), the place where the vetor L. longipalpis takes its blood meal, compared with those from lymph node (6.3/mm² x 8.3/mm²) and viscera (1.4/mm²x 5.3/mm²).
Assuntos
Doenças do Cão/parasitologia , Leishmania infantum/isolamento & purificação , Leishmaniose Visceral/veterinária , Linfonodos/parasitologia , Vísceras/parasitologia , Animais , Brasil , Doenças do Cão/transmissão , Cães , Feminino , Imuno-Histoquímica , Insetos Vetores , Leishmaniose Visceral/parasitologia , Leishmaniose Visceral/transmissão , Masculino , Psychodidae , Pele/parasitologiaRESUMO
Canine visceral leishmaniasis (CVL) is recognizable by characteristic signs of disease and is highly lethal. The infection, however, may be quite inapparent in some seropositive dogs, and this has raised the polemic question as to whether or not such animals can be a source of infection for Lutzomyia longipalpis, the vector of American visceral leishmaniasis (AVL). In this study we have examined 51 dogs with acute CVL from an AVL area in Pará State, northern Brazil, and compared the parasite density, amastigotes of Leishmania (L.) infantum chagasi, in the skin, lymph node and viscera of symptomatic with that of nine asymptomatic but seropositive dogs (IFAT-IgG). Post-mortem biopsy fragments of these tissues were processed by immunohistochemistry, using a polyclonal antibody against Leishmania sp. The X² and Mann Whitney tests were used to evaluate the means of infected macrophage density (p < 0.05). There was no difference (p > 0.05) in the skin (10.7/mm² x 15.5/mm²) and lymph node (6.3/mm² x 8.3/mm²), between asymptomatic and symptomatic dogs, respectively. It was higher (p < 0.05), however, in the viscera of symptomatic (5.3/mm²) than it was in asymptomatic (1.4/mm²) dogs. These results strongly suggest that asymptomatic or symptomatic L. (L.) i. chagasi-infected dogs can serve as a source of infection, principally considering the highest (p < 0.05) parasite density from skin (10.7/mm² x 15.5/mm²), the place where the vetor L. longipalpis takes its blood meal, compared with those from lymph node (6.3/mm² x 8.3/mm²) and viscera (1.4/mm²x 5.3/mm²).
A leishmaniose visceral canina (LVC) é reconhecida pelas características clínicas da doença e é altamente letal. A infecção, entretanto, pode ser totalmente assintomática em alguns cães soropositivos, o que tem levantado questão polêmica sobre a possibilidade desses animais, serem ou não uma fonte importante da infecção para o flebotomíneo, Lutzomyia longipalpis, o principal vetor da leishmaniose visceral americana (LVA). Neste estudo foram examinados 51 cães com LVC aguda, provenientes de área endêmica de LVA no Estado do Pará, Brasil, e a carga parasitária, formas amastigotas de Leishmania (L.) infantum chagasi, na pele, linfonodo poplíteo e vísceras (fígado e baço) foi comparada com a de nove cães assintomáticos soropositivos (IFAT-IgG). Fragmentos de biópsia desses tecidos obtidos post-mortem foram processados para análise através de imunohistoquímica, usando um anticorpo policlonal contra Leishmania sp. Os testes do Qui-quadrado (X²) e Mann Whitney foram usados para avaliar as médias da densidade de macrófagos infectados (p < 0,05). Os resultados mostraram que não houve diferença (p > 0,05) na densidade de macrófagos infectados da pele (10,7/mm² x 15,5/mm²) e do linfonodo (6,3/mm² x 8,3/mm²) entre cães assintomáticos e sintomáticos. Entretanto, a densidade de macrófagos infectados da víscera de cães sintomáticos (5,3/mm²) foi maior (p < 0,05) que a de cães assintomáticos (1,4/mm²). Estes resultados sugerem, fortemente, que cães naturalmente infectados por L. (L.) i. chagasi, assintomáticos ou sintomáticos, podem servir como fonte de infecção, principalmente, considerando-se que a densidade de macrófagos infectados da pele (10,7/mm² x 15,5/mm²), local onde o flebotomíneo vetor Lu. longipalpis realiza a hematofagia, foi maior (p < 0,05) que as do linfonodo (6,3/mm² x 8.3/mm²) e vísceras (1,4/mm²x 5,3/mm²).
Assuntos
Animais , Cães , Feminino , Masculino , Doenças do Cão/parasitologia , Leishmania infantum/isolamento & purificação , Leishmaniose Visceral/veterinária , Linfonodos/parasitologia , Vísceras/parasitologia , Brasil , Doenças do Cão/transmissão , Imuno-Histoquímica , Insetos Vetores , Leishmaniose Visceral/parasitologia , Leishmaniose Visceral/transmissão , Psychodidae , Pele/parasitologiaRESUMO
The purpose of this study was to describe the reproductive profile and frequency of genital infections among women living in the Serra Pelada, a former mining village in the Pará state, Brazil. A descriptive study of women living in the mining area of Serra Pelada was performed in 2004 through interviews that gathered demographics and clinical data, and assessed risk behaviors of 209 randomly-selected women. Blood samples were collected for rapid assay for HIV; specimens were taken for Pap smears and Gram stains. Standard descriptive statistical analyses were performed and prevalence was calculated to reflect the relative frequency of each disease. Of the 209 participants, the median age was 38 years, with almost 70% having less than four years of education and 77% having no income or under 1.9 times the minimum wage of Brazil. About 30% did not have access to health care services during the preceding year. Risk behaviors included: alcohol abuse, 24.4%; illicit drug abuse, 4.3%; being a sex worker, 15.8%; and domestic violence, 17.7%. Abnormal Pap smear was found in 8.6%. Prevalence rates of infection were: HIV, 1.9%; trichomoniasis, 2.9%; bacterial vaginosis, 18.7%; candidiasis, 5.7%; Chlamydial-related cytological changes, 3.3%; and HPV-related cytological changes, 3.8%. Women living in this mining area in Brazil are economically and socially vulnerable to health problems. It is important to point out the importance of concomitant broader strategies that include reducing poverty and empowering women to make improvements regarding their health.
Assuntos
Doenças dos Genitais Femininos/epidemiologia , Saúde da Mulher , Adulto , Brasil , Feminino , Humanos , Pessoa de Meia-Idade , Medicina Reprodutiva , Adulto JovemRESUMO
The purpose of this study was to describe the reproductive profile and frequency of genital infections among women living in the Serra Pelada, a former mining village in the Pará state,Brazil. A descriptive study of women living in the mining area of Serra Pelada was performed in 2004 through interviews that gathered demographics and clinical data, and assessed risk behaviors of 209 randomly-selected women. Blood samples were collected for rapid assay for HIV; specimens were taken for Pap smears and Gram stains. Standard descriptive statistical analyseswere performed and prevalence was calculated to reflect the relative frequency of each disease. Of the 209 participants, the median age was 38 years, with almost 70% having less thanfour years of education and 77% having no income or under 1.9 times the minimum wage of Brazil. About 30% did not have access to health care services during the preceding year. Risk behaviors included: alcohol abuse, 24.4%; illicit drug abuse, 4.3%; being a sex worker, 15.8%; and domestic violence, 17.7%. Abnormal Pap smear was found in 8.6%. Prevalence rates of infection were: HIV, 1.9%; trichomoniasis, 2.9%; bacterial vaginosis, 18.7%; candidiasis, 5.7%;Chlamydial-related cytological changes, 3.3%; and HPV-related cytological changes, 3.8%. Women living in this mining area in Brazil are economically and socially vulnerable to health problems. It is important to point out the importance of concomitant broader strategies that include reducing poverty and empowering women to make improvements regarding their health.
El propósito de este estudio fue describir el perfil reproductivo y la frecuencia de infecciones genitales en mujeres que viven en la población minera Serra Pelada en el Estado de Pará, Brasil. Se realizó un estudio descriptivo de las mujeres que vivían en la zona minera de Serra Pelada en 2004 mediante entrevistas en las que se recabarondatos demográficos y clínicos y se examinaron las conductas de riesgo de 209 mujeres seleccionadas aleatoriamente. Se tomaron muestras de sangre para pruebas rápidas de detección de anticuerpos contra el VIH y muestras de tejido para análisis citológico y tinción de Gram. Se calculó la prevalencia y se utilizaron pruebas estadísticas descriptivas estándares para caracterizar la frecuencia relativa de cada enfermedad. La mediana de la edad de las 209 participantes fue de 38 años; 70% tenía menos de cuatro años de escolaridad y 77% no tenía ingresos o estos eran inferiores a 1,9 veces elsalario mínimo en Brasil. Alrededor de 30% no tuvo acceso a servicios de salud durante el año previo. Entre las conductas de riesgo estaban: consumo de bebidas alcohólicas (24,4%) y de drogas ilícitas (4,3%), ser trabajadora sexual (15,8%) y violencia doméstica (17,7%). Se encontraron resultados anormales a la prueba citológica en8,6% de las participantes. Las prevalencias de infección fueron: 1,9% de VIH, 2,9% de tricomoniasis, 18,7% de vaginosis bacteriana y 5,7% de candidiasis; 3,3% presentó alteracionescitológicas asociadas con la infección por clamidia y 3,8% alteraciones citológicas asociadas con el virus de papiloma humano. Las mujeres de esta zona minera de Brasil son económica y socialmente vulnerables a problemas de salud. Es importante señalar la importancia de estrategias concomitantes más amplias que abarquen la reducción de la pobreza y el empoderamiento de las mujeres para lograr mejoras en su salud.
Assuntos
Adulto , Feminino , Humanos , Pessoa de Meia-Idade , Adulto Jovem , Doenças dos Genitais Femininos/epidemiologia , Saúde da Mulher , Brasil , Medicina Reprodutiva , Adulto JovemRESUMO
Molecular characterization of Cryptosporidium spp.oocysts in clinical samples is useful for public health since it allows the study of sources of contamination as well as the transmission in different geographical regions. Although widely used in developed countries, in Brazil it is restricted to academic studies, mostly using commercial kits for the extraction of genomic DNA, or in collaboration with external reference centers, rendering the method expensive and limited. The study proposes the application of the modifications recently introduced in the method improving feasibility with lower cost. This method was efficient for clinical samples preserved at -20 °C for up to six years and the low number of oocysts may be overcomed by repetitions of extraction.
A caracterização molecular de oocistos de Cryptosporidium spp. em amostras clínicas é útil à saúde pública, pois permite estudo das fontes de contaminação e a transmissão em determinadas regiões geográficas. Apesar de largamente utilizada em países desenvolvidos, no Brasil está restrita aos estudos acadêmicos, na maioria utilizando kits comerciais para extração do DNA genômico, ou em colaborações com centros de referência externos, o que torna o método caro e limitado. Este estudo propõe a introdução de modificações nos métodos existentes para melhorar a viabilidade e baixar custos. O método proposto foi eficiente em amostras clínicas preservadas a -20 °C por até seis anos e o baixo número de oocistos pode ser contornado por replicadas extrações de DNA.
Assuntos
Animais , Humanos , Criptosporidiose/diagnóstico , Cryptosporidium/genética , DNA de Protozoário/isolamento & purificação , Fezes/parasitologia , Oocistos , Protocolos Clínicos , Cryptosporidium/isolamento & purificação , Reação em Cadeia da Polimerase , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
The aim of this investigation was to evaluate clinicopathologic and immunohistochemical characteristics of synchronous primary gastric adenocarcinomas. Immunohistochemistry for p53 (suppressor pathway) and for hMLH1, hMSH2, and hMSH6 (mutator pathway) was performed using ABC-technique amplification by biotinylated tyramide. Synchronous primary gastric adenocarcinomas were detected in 19/553 (3.43%) of the patients. The tumors were localized in distal stomach in 22, body in 14, and proximal in five. There was a predominance of intestinal type in the group of synchronic tumors compared to the solitary lesions, 73.2 vs 37.3%, p = 0.001. Synchronous neoplasias were diagnosed in earlier stage than solitary neoplasias, T1-T2 = 60.9% vs T1-T2 = 28.4%, p = 0.0001; and N0 = 68.4% vs N0 = 26.2%, p = 0.001. p53 was detected in 52.6% of the patients with synchronous tumors. Altered hMLH1 immunoexpression occurred in 26.3% of the patients and hMSH6 in 5.3%. hMSH2 immunoreactivity was positive in all tumors. p53 was solely detected in 17 tumors, while hMLH1 was altered in 10/24 negative p53 tumors, p = 0.01. Synchronous gastric adenocarcinomas presented higher frequency of intestinal type and early gastric cancer in comparison to solitary gastric cancer. Two routes of carcinogenesis, mutator, and suppressor appear to be involved independently in the development of synchronous tumors.
Assuntos
Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Neoplasias Primárias Múltiplas/metabolismo , Neoplasias Primárias Múltiplas/patologia , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Proteínas Adaptadoras de Transdução de Sinal/análise , Idoso , Enzimas Reparadoras do DNA/análise , Proteínas de Ligação a DNA/análise , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Proteína 1 Homóloga a MutL , Proteína 2 Homóloga a MutS/análise , Proteínas Nucleares/análise , Proteína Supressora de Tumor p53/análiseRESUMO
UNLABELLED: Three subtypes of enterochromaffin-like cell tumors (carcinoids) have been described: type I, associated with chronic atrophic gastritis; type II, multiple endocrine neoplasia 1 and Zollinger-Ellison syndrome; and type III, sporadic tumors. OBJECTIVES: (i) To investigate the immunoexpression of Ki-67, p53 and Bcl-2 proteins in enterochromaffin-like cell (carcinoid) tumors and (ii) to evaluate the prognostic value of these markers. METHODS: Fifty-four samples from 21 patients with gastric carcinoid tumors were sectioned and immunostained using avidin-biotin peroxidase method. RESULTS: The mean age was 62.2+/-11.4 years (36-83 years-old) and 13 (61.9%) were women. Type I lesions were detected in 61.9% and type III in 38.1%. Tumors were single in 10 (47.6%) and were multiple and/or multicentric in 11 (52.4%). Nuclear p53 immunoreactivity was observed in 6/21 patients (28.6%), and all of them were type III tumors (6/8), compared with no p53 expression in type I (0/13), P=0.0002. p53 expression was also associated with high degree of cell proliferation (Ki-67-positive nuclear cells), P=0.00001. Bcl-2 expression was either unreactive or weakly positive in all tumor lesions. The mean follow-up period was 50.4 months (SD=45.2), varying from 6 to 144 months. Overall survival time of patients with positive p53 expression and high proliferative rate was significantly lower than that of negative patients (14.4 vs 123 months, P=0.0007). CONCLUSIONS: (i) p53 immunoexpression associated with high proliferative rate was useful to distinguish between type I and type III gastric carcinoid tumors and (ii) these markers were able to predict a shorter survival.
Assuntos
Biomarcadores Tumorais/metabolismo , Tumor Carcinoide/diagnóstico , Neoplasias Gástricas/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Tumor Carcinoide/patologia , Proliferação de Células , Feminino , Gastroscopia , Humanos , Técnicas Imunoenzimáticas , Antígeno Ki-67/metabolismo , Masculino , Pessoa de Meia-Idade , Prognóstico , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Estudos Retrospectivos , Neoplasias Gástricas/patologia , Análise de Sobrevida , Proteína Supressora de Tumor p53/metabolismoRESUMO
Experimental animal models have been used for the study of the physiopathogenesis of leishmaniasis, on some occasions with success, while in other situations such as bone alterations that accompany tegumentary leishmaniasis, especially in diffuse cutaneous form (DCL), the mechanisms are still unknown. In the present study, we determined these alterations in an animal model susceptible to Leishmania (L) amazonensis. Amastigotes of L. (L) amazonensis isolated from patients with diffuse cutaneous leishmaniasis (DCL) were inoculated into the hind paws of eight BALB/c mice, macroscopic and histopathological aspects were analyzed. After 90 and 120 days of evolution, histopathological analysis demonstrated a mononuclear cell infiltrate rich in plasma cells and intense parasitism of intra- and extra-medullary macrophages, with areas of bone necrosis and discrete involvement of cartilaginous tissue. The results show that the inflammatory process developed during L. (L) amazonensis infection might cause bone tissue destruction and secondarily affect the joints.
Assuntos
Leishmania/crescimento & desenvolvimento , Leishmaniose Tegumentar Difusa/patologia , Osteomielite/parasitologia , Animais , Modelos Animais de Doenças , Membro Posterior/parasitologia , Membro Posterior/patologia , Histocitoquímica , Humanos , Leishmaniose Tegumentar Difusa/imunologia , Leishmaniose Tegumentar Difusa/parasitologia , Macrófagos/imunologia , Macrófagos/parasitologia , Macrófagos/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Osteomielite/imunologia , Osteomielite/patologiaRESUMO
BACKGROUND: Pneumoperitoneum may be responsible for ultra-structural alterations in the mesothelium during laparoscopy. To characterize the effect of pneumoperitoneum on the mesothelial cells with CO(2) and compressed air; and to compare to laparotomy and control group (anesthesia only). MATERIALS AND METHODS: Forty C-57 mice were divided in four groups of 10 animals each: CO(2), air, laparotomy, and control group. The animals were submitted to pneumoperitoneum at 8 mmHg during 30 min (CO(2) or compressed air). Five animals of each group were sacrificed 2 and 24 h after the procedure. Fragments of parietal peritoneum were collected and processed for scanning electron microscopy. RESULTS: Control group revealed uninterrupted mesothelial cells, without any evidence of cellular limits; close contact between the cells; absence of intercellular clefts and presence of microvilli. In the laparotomy group, similar results to the control group, with decreased microvilli were noted. Air pneumoperitoneum was associated with alterations in the morphology of the mesothelial cells, clear cellular limits, and cells with spherical and fusiforme formats. CO(2) pneumoperitoneum showed mesothelial cells with clear cellular limits, predominantly spherical cellular format, and intercellular clefts that allowed the visualization of the exposed basal membrane. These alterations were more intense after 24 h. There was a statistical significance between CO(2) group (2 and 24 h) compared to the control group and laparotomy for cellular limits, intercellular clefts and microvilli, P < 0.0001. CONCLUSIONS: Pneumoperitoneum causes damage in the mesothelial ultra-structure, which differs from the laparotomy group. CO(2) pneumoperitoneum is more harmful to the mesothelium than the air.
Assuntos
Laparoscopia/efeitos adversos , Peritônio/patologia , Pneumoperitônio/etiologia , Pneumoperitônio/patologia , Ar , Animais , Dióxido de Carbono , Modelos Animais de Doenças , Epitélio/patologia , Epitélio/ultraestrutura , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microscopia Eletrônica de Varredura , Peritônio/ultraestrutura , Pressão , Estresse MecânicoRESUMO
Leishmania (Leishmania) amazonensis has for some time been considered as the causative agent of two distinct forms of American cutaneous leishmaniasis (ACL): localized cutaneous leishmaniasis (LCL), and anergic diffuse cutaneous leishmaniasis (ADCL). Recently, a new intermediate form of disease, borderline disseminated cutaneous leishmaniasis (BDCL), was introduced into the clinical spectrum of ACL caused by this parasite, and in this paper we record the clinical, histopathological, and immunological features of eight more BDCL patients from Brazilian Amazonia, who acquired the disease in the Pará state, North Brazil. Seven of them had infections of one to two years' evolution and presented with primary skin lesions and the occurrence of metastases at periods varying from six to 12 months following appearance of the first lesion. Primary skin lesions ranged from 1-3 in number, and all had the aspect of an erythematous, infiltrated plaque, variously located on the head, arms or legs. There was lymphatic dissemination of infection, with lymph node enlargement in seven of the cases, and the delayed hypersensitivity skin-test (DTH) was negative in all eight patients prior to their treatment. After that, there was a conversion of DTH to positive in five cases re-examined. The major histopathological feature was a dermal mononuclear infiltration, with a predominance of heavily parasitized and vacuolated macrophages, together with lymphocytes and plasma cells. In one case, with similar histopathology, the patient had acquired his infection seven years previously and he presented with the largest number of disseminated cutaneous lesions. BDCL shows clinical and histopathological features which are different from those of both LCL and ADCL, and there is a good prognosis of cure which is generally not so in the case of frank ADCL.
Assuntos
Leishmania mexicana , Leishmaniose Cutânea , Adolescente , Adulto , Animais , Biópsia , Criança , Humanos , Leishmaniose Cutânea/tratamento farmacológico , Leishmaniose Cutânea/imunologia , Leishmaniose Cutânea/patologia , Masculino , Pessoa de Meia-IdadeRESUMO
Leishmania (Leishmania) amazonensis has for some time been considered as the causative agent of two distinct forms of American cutaneous leishmaniasis (ACL): localized cutaneous leishmaniasis (LCL), and anergic diffuse cutaneous leishmaniasis (ADCL). Recently, a new intermediate form of disease, borderline disseminated cutaneous leishmaniasis (BDCL), was introduced into the clinical spectrum of ACL caused by this parasite, and in this paper we record the clinical, histopathological, and immunological features of eight more BDCL patients from Brazilian Amazonia, who acquired the disease in the Pará state, North Brazil. Seven of them had infections of one to two years' evolution and presented with primary skin lesions and the occurrence of metastases at periods varying from six to 12 months following appearance of the first lesion. Primary skin lesions ranged from 1-3 in number, and all had the aspect of an erythematous, infiltrated plaque, variously located on the head, arms or legs. There was lymphatic dissemination of infection, with lymph node enlargement in seven of the cases, and the delayed hypersensitivity skin-test (DTH) was negative in all eight patients prior to their treatment. After that, there was a conversion of DTH to positive in five cases re-examined. The major histopathological feature was a dermal mononuclear infiltration, with a predominance of heavily parasitized and vacuolated macrophages, together with lymphocytes and plasma cells. In one case, with similar histopathology, the patient had acquired his infection seven years previously and he presented with the largest number of disseminated cutaneous lesions. BDCL shows clinical and histopathological features which are different from those of both LCL and ADCL, and there is a good prognosis of cure which is generally not so in the case of frank ADCL.
Assuntos
Humanos , Animais , Masculino , Criança , Adolescente , Adulto , Pessoa de Meia-Idade , Leishmania mexicana , Leishmaniose Cutânea , Biópsia , Leishmaniose Cutânea/diagnóstico , Leishmaniose Cutânea/tratamento farmacológico , Leishmaniose Cutânea/imunologia , Leishmaniose Cutânea/patologiaRESUMO
The wide variety of Leishmania species responsible for human American cutaneous leishmaniasis combined with the immune mechanisms of the host results in a large spectrum of clinical, histopathological, and immunopathological manifestations. At the middle of this spectrum are the most frequent cases of localized cutaneous leishmaniasis (LCL) caused by members of the subgenera Leishmania and Viannia, which respond well to conventional therapy. The two pathogenicity extremes of the spectrum generally recognized are represented at the hypersensitivity pole by mucocutaneous leishmaniasis (MCL) and at the hyposensitivity pole by anergic diffuse cutaneous leishmaniasis (ADCL). Following the present study on the clinical, histopathological and immunopathological features of cutaneous leishmaniasis in Amazonian Brazil, we propose the use of the term "borderline disseminated cutaneous leishmaniasis" for the disseminated form of the disease, due to parasites of the subgenera Leishmania and Viannia, which might be regarded as intermediate between LCL and the extreme pathogenicity poles MCL and ADCL.