Lysosomal Acid Lipase Deficiency: Report of Five Cases across the Age Spectrum.

Lysosomal acid lipase (LAL) deficiency is an autosomal recessive lysosomal storage disorder caused by mutations in the LIPA gene that leads to premature organ damage and mortality. We present retrospective data from medical records of 5 Brazilian patients, showing the broad clinical spectrum of the disease.

Caracterização histopatológica da esofagite eosinofílica em crianças e adolescentes / Histological features of eosinophilic esophagitis in children and adolescents

ABSTRACT BACKGROUND: Eosinophilic esophagitis is an emerging disease featured by eosinophilic esophageal infiltrate not responsive to proton pump inhibitors. OBJECTIVE: To characterize histological features of children and adolescents with eosinophilic esophagitis. METHODS: Cross-sectional study in a tertiary hospital. Biopsies from each esophageal third from 14 patients (median age 7 years) with eosinophilic esophagitis were evaluated. Histological features evaluated included morphometry of esophageal epithelium, esophageal density (per high power field), extracellular eosinophilic granules, eosinophilic microabscesses, surface disposition of eosinophils, epithelial desquamation, peripapillary eosinophilia, basal layer hyperplasia and papillary elongation. RESULTS: Several patients presented a normal esophageal macroscopy in the upper digestive endoscopy (6, 42.8%), and the most common abnormality were vertical lines (7, 50%) and whitish spots over esophageal mucosa (7, 50%). Basal layer hyperplasia was observed in 88.8%, 100% e 80% of biopsies from proximal, middle and lower esophagus, respectively (P=0.22). Esophageal density ranges from 0 to more than 50 per hpf. Extracellular eosinophilic granules (70%-100%), surface disposition of eosinophils (60%-93%), epithelial desquamation (60%-100%), peripapillary eosinophilia (70%-80%) were common, but evenly distributed among each esophageal third. Just one patient did not present eosinophils in the lower third, four in the middle third and four in the upper esophageal third. CONCLUSION: In the absence of hypereosinophilia, other histological features are present in eosinophilic esophagitis and may contribute to diagnosis. Eosinophilic infiltrate is focal, therefore multiple biopsies are needed for diagnosis.

RESUMO CONTEXTO: Esofagite eosinofílica é uma doença emergente caracterizada por infiltrado eosinofílico esofágico não responsivo a inibidores de bomba de prótons. OBJETIVO: Caracterizar os achados histopatológicos de uma coorte de crianças e adolescentes com diagnóstico de esofagite eosinofílica. MÉTODOS: Estudo transversal conduzido em hospital terciário. Biópsias de terços proximal, médio e distal de 14 pacientes (idade mediana 7 anos) com diagnóstico de esofagite eosinofílica. Estudo morfométrico e variáveis histológicas analisadas em fragmentos de biópsias nos terços esofágicos: contagem de eosinófilos/CGA, grânulos eosinofílicos extracelulares, microabscessos eosinofílicos, disposição superficial de eosinófilos, descamação epitelial, eosinofilia peripapilar, hiperplasia da camada basal e alongamento de papilas. RESULTADOS: Vários pacientes apresentaram aspecto macroscópico normal da mucosa esofágica à endoscopia (6, 42.8%), e a anormalidade mais comumente observada foi linhas verticais (7, 50%) e exsudato branco (7, 50%). Hiperplasia da camada basal foi observada em 88,8%, 100% e 80% das biópsias do terço proximal, médio e distal respectivamente (P=0,22); contagem de eosinófilos nos terços variou de 0 a ≥50/CGA, grânulos eosinofílicos extracelulares (70%-100%), disposição superficial de eosinófilos (60%-93%), descamação epitelial (60%-100%), eosinofilia peripapilar (70%-80%), sem diferença estatística entre os terços esofágicos. Ausência de eosinofilia ocorreu raramente em terço distal (uma do distal, quatro do proximal, quatro do médio). CONCLUSÃO: Na ausência de hipereosinofilia, outros achados histopatológicos de inflamação eosinofílica estão presentes. A infiltração eosinofílica apresentou caráter focal, sugerindo-se a realização de múltiplas biópsias de diversos segmentos.

HISTOLOGICAL FEATURES OF EOSINOPHILIC ESOPHAGITIS IN CHILDREN AND ADOLESCENTS.

BACKGROUND: Eosinophilic esophagitis is an emerging disease featured by eosinophilic esophageal infiltrate not responsive to proton pump inhibitors. OBJECTIVE: To characterize histological features of children and adolescents with eosinophilic esophagitis. METHODS: Cross-sectional study in a tertiary hospital. Biopsies from each esophageal third from 14 patients (median age 7 years) with eosinophilic esophagitis were evaluated. Histological features evaluated included morphometry of esophageal epithelium, esophageal density (per high power field), extracellular eosinophilic granules, eosinophilic microabscesses, surface disposition of eosinophils, epithelial desquamation, peripapillary eosinophilia, basal layer hyperplasia and papillary elongation. RESULTS: Several patients presented a normal esophageal macroscopy in the upper digestive endoscopy (6, 42.8%), and the most common abnormality were vertical lines (7, 50%) and whitish spots over esophageal mucosa (7, 50%). Basal layer hyperplasia was observed in 88.8%, 100% e 80% of biopsies from proximal, middle and lower esophagus, respectively (P=0.22). Esophageal density ranges from 0 to more than 50 per hpf. Extracellular eosinophilic granules (70%-100%), surface disposition of eosinophils (60%-93%), epithelial desquamation (60%-100%), peripapillary eosinophilia (70%-80%) were common, but evenly distributed among each esophageal third. Just one patient did not present eosinophils in the lower third, four in the middle third and four in the upper esophageal third. CONCLUSION: In the absence of hypereosinophilia, other histological features are present in eosinophilic esophagitis and may contribute to diagnosis. Eosinophilic infiltrate is focal, therefore multiple biopsies are needed for diagnosis.

Diagnosis of Helicobacter pylori infection by means of reduced-dose ¹³C-urea breath test and early sampling of exhaled breath.

OBJECTIVE: The aim of this study was to evaluate the accuracy of reduced-dose ¹³C-urea breath test (¹³C-UBT) and early sampling of exhaled breath for the detection of Helicobacter pylori infection in children and adolescents. METHODS: Patients up to 20 years old that underwent upper gastrointestinal endoscopy with gastric biopsies were included. The ¹³C-UBT was performed after a 4-hour fasting period with 4 points of collection: baseline (T0), and at 10, 20, and 30 minutes (T10, T20, and T30) after ingestion of 25 mg ¹³C-urea diluted in 100 mL of apple juice. The infection status was defined through 3 invasive methods, and a patient was considered infected with a positive culture or concomitant positive histology and rapid urease test. The absence of infection was defined by all negative histology, rapid urease test, and culture. Analysis of exhaled breath samples was performed with an isotope-selective infrared spectrometer. A receiver-operating characteristic curve analysis was done to define cutoff delta over baseline (DOB) values. RESULTS: A total of 129 patients between the ages of 2.1 and 19 years (median 11.6 years; mean age ± standard deviation 11.5 ± 3.8 years; F:M 85:44) were included. The prevalence of infection was 41.1%. The sensitivity (S) and specificity (Sp) were at T10 (cutoff DOB 2.55‰), S 94.7% (95% confidence interval [CI] 90.9-98.5) and Sp 96.8% (95% CI 93.4-100); at T20 (DOB 2.5‰), S 96.2% (95% CI 92.9-99.5) and Sp 96.1% (95% CI 93.7-99.8); and at T30 (DOB 1.6‰), S 96.2% (95% CI 92.9-99.5) and Sp 94.7% (95% CI 90.8-98.6). CONCLUSIONS: Low-dose ¹³C-UBT with early sampling is accurate for diagnosing H pylori infection in children and adolescents.

Morphometrical and immunohistochemical study of intrahepatic bile ducts in biliary atresia.

BACKGROUND: Extrahepatic biliary atresia results from a progressive destruction of the bile ducts by an inflammatory fibrosing process which leads ultimately to cirrhosis of biliary type. The etiology of the disorder remains unknown. The histological features include cholestasis, ductular proliferation, eventual loss of intrahepatic bile ducts, and ducts with primitive embryonic shape (ductal plate malformation). PURPOSE: To examine the morphological changes of the biliary intrahepatic ducts, we aimed at investigating the cell proliferation and the diameter of the interlobular bile ducts in extrahepatic biliary atresia, and in normal liver children. METHODS: Liver samples from 35 patients with biliary atresia and 10 from control normal children were used. Immunoexpression of cytokeratin 19 was evaluated and a double-staining procedure was performed with cytokeratin 8/proliferating cell nuclear antigen. The stereological measurements of the intrahepatic bile ducts diameter were evaluated by a computerized system of image analysis. RESULTS: The patterns of intrahepatic cholangiopathy in biliary atresia were obstructive features (42.86%), paucity of intrahepatic bile ducts (20%), ductal plate malformation (28.57%), and ductal plate malformation associated with paucity of intrahepatic bile ducts (8.57%). The average external diameter of interlobular bile ducts in biliary atresia was smaller than that of the control infant livers. Among the four patterns of biliary atresia cholangiopathies, those associated with ductopenia showed the smallest bile duct diameter. There was a negative correlation between the bile duct to portal space ratio and the age of the child at the time of Kasai portoenterostomy. Only in biliary atresia are the bile duct cells stained with proliferating cell nuclear antigen. CONCLUSION: (i) In biliary atresia, both ductular metaplasia and ductular proliferation were observed; (ii) biliary atresia associated with ductopenia showed narrowing of interlobular ducts, probably as a consequence of degeneration with atrophy and fibrosis.

Síndrome megabexiga microcólon hipoperistaltismo intestinal: sobrevida até a adolescência / Megacystis-microcolon-intestinal hypoperistalsis syndrome: survival to adolescence

A síndrome megabexiga microcólon hipoperistaltismo intestinal (SMMHI) é uma doença rara, de provável herança genética, com prognóstico reservado, já no primeiro ano de vida. Relata-se um caso de SMMHI em criança do sexo feminino, atualmente com 16 anos de idade, com o objetivo de ressaltar o prognóstico favorável, não esperado, com sobrevida até a adolescência, de uma síndrome rara. Chama-se a atenção para a possibilidade de estabelecer o diagnóstico no período perinatal e para a importância de realizar o estudo urodinâmico.

Intraepithelial lymphocytes in duodenum from Brazilian adolescents with type 1 diabetes. Influence of Helicobacter pylori.

BACKGROUND: An increased number of intraepithelial lymphocytes (IELs) can be the only histological feature in early stages of celiac disease (CD). This is also presented in duodenum of patients with Helicobacter pylori-associated gastritis and in autoimmune diseases. Because CD is frequently associated with type 1 diabetes mellitus, we analyzed the density of IELs in the distal duodenum of non-celiac diabetic patients associated or not with H.pylori infection. METHODS: IEL density and the presence of H.pylori were determined in biopsies of the distal duodenum and gastric antrum and body obtained from Brazilian diabetic adolescents who were negative for anti-human tissue transglutaminase and anti-endomysial. The results were compared with the histological findings of gastric and duodenal biopsies obtained from non-diabetic older children and adolescents. RESULTS: H.pylori was detected in 33.3% of diabetic patients and in 56.7% of the control group. No association was observed between the presence of H.pylori and an increased lymphocyte density in the distal duodenum in either group. Diabetic patients presented a duodenal IEL density similar to that of the control group. Lymphocytic gastritis was not identified in any of the biopsies analyzed. CONCLUSIONS: The density of IELs in the distal duodenum of diabetic adolescents did not differ from that observed in older children and adolescents without this autoimmune disease. H.pylori infection, which is frequent among adolescents from developing countries, did not modify lymphocyte density in the distal duodenum in the absence of lymphocytic gastritis.

Osteossarcomas humanos de alto grau: imunoexpressão de p53, erb-2 e P-glicoproteína, e correlação com o parâmetro anaplasia / P-glycoprotein, erb2 and p53 expression in high-grade human osteosarcomas and their correlation with anaplasia

INTRODUÇÃO: Osteossarcoma (OS), o mais freqüente tumor primário maligno do osso, tem comportamento local agressivo e alto índice de disseminação sistêmica. Os eventos que permitem o crescimento e a disseminação tumoral ainda permanecem controversos. Os estudos sobre a carcinogênese e a progressão dessa neoplasia, com base na imunoexpressão de c-erb-B2, P-glicoproteína (P-gp) e p53, apresentam resultados conflitantes acerca do real valor prognóstico e suas correlações com parâmetros histológicos. A anaplasia, em neoplasias na infância, constitui parâmetro histológico de agressividade tumoral e quimiorresistência. Nos OS primários ou metastáticos, seu significado permanece controverso. Por outro lado, em outras neoplasias humanas, a expressão do c-erb-B2 relaciona-se com p53, grau nuclear e outros parâmetros de agressividade. OBJETIVO: Avaliar a imunoexpressão de p53, c-erb-B2 e P-gp em OS, correlacionando os parâmetros entre si e com a presença de anaplasia. MÉTODO: O estudo incluiu 96 biópsias pré-quimioterapia de pacientes com OS de alto grau, diagnosticados entre 1991 e 2000. A pesquisa imuno-histoquímica de p53, P-gp e c-erb-B2 foi feita pela técnica da estreptoavidina-biotina-peroxidase. Foram considerados positivos os casos onde havia imunoexpressão em 10 por cento ou mais das células neoplásicas. Somente colorações membranosa (para cerb-B2 e P-gp) e nuclear (para p53) foram consideradas positivas. Anaplasia foi definida como no tumor de Wilms, sendo considerada presente ou ausente. RESULTADOS: Anaplasia pôde ser avaliada em 82/96 casos, estando presente em 29 (35,36 por cento). Imunoexpressão de p53 foi detectada em 25 dos 60 casos (36,23 por cento); de P-gp, em 30 dos 73 casos (41,1 por cento); e de c-erb-B2, em 22 dos 55 casos (40 por cento). Os resultados demonstraram associação entre as imunoexpressões de c-erb-B2 e p53 (p = 0,042), p53 e o parâmetro anaplasia (p = 0,015), anaplasia e Pg (p = 0.034) CONCLUSÕES: A imunoexpressão...

BACKGROUND: Osteosarcomas (OS), the most frequent primary malignant bone tumors, have aggressive local behavior and high rate of metastatization. The events that allow tumor growth and dissemination are still controversial. The studies about carcinogenesis and tumor progression in this neoplasia, which are based on c-erb-B2, P-glycoprotein (P-gp) and p53 immunoexpression, show conflicting results as to the real prognostic value and its correlations with histological parameters. Anaplasia in childhood neoplasias is a histological parameter of tumor aggressiveness and chemoresistance. In primary or metastatic OS, its meaning remains controversial. On the other hand, in other human neoplasias, c-erb-B2 expression is associated with p53, nuclear grade and other aggressiveness parameters. OBJECTIVE: The aim of the present study was to evaluate p53, c-erb-B2 and P-gp immunoexpression in OS, correlating the parameters with the presence of anaplasia. METHODS: This study included 96 pre-chemotherapy biopsies in patients with high-grade OS diagnosed between 1991 and 2000. The immunohistochemical evaluation of p53 and c-erb-B2 was carried out with the streptavidin-biotin-peroxidase technique. Cases were considered positive when there was immunoexpression in 10 percent or more neoplastic cells. Only membrane staining (for c-erb-B2 and P-gp), and nuclear staining (for p53) were considered positive. Anaplasia was defined as Wilms' tumor, and considered present or absent. RESULTS: Anaplasia was present in 29 out of 82 cases (35.36 percent); p53 immunoexpression was detected in 25 out of 60 cases (36.23 percent); P-gp, in 30 out of 73 cases (41.1 percent); and c-erb-B2, in 22 out of 55 cases (40 percent). The results demonstrated an association between c-erb-B2 and p53 immunoexpression (p = 0.042), p53 and parameter of anaplasia (p = 0.015), anaplasia and P-gp (p = 0.034). CONCLUSIONS: The p53, c-erb-B2 and P-gp immunoexpression is relatively frequent in high-grade, metastic...

Gastric emptying of solids is slower in functional dyspepsia unrelated to Helicobacter pylori infection in female children and teenagers.

AIM: To evaluate gastric emptying of solids in children and adolescents with functional dyspepsia with and without Helicobacter pylori infection. PATIENTS AND METHODS: The study included 27 female patients (mean age 13.38 +/- 2.81 y) with functional dyspepsia according to Rome II criteria who were selected after upper gastrointestinal endoscopy found no major mucosal abnormality. Fragments were collected from the esophagus, gastric antrum, and gastric body for histological examination and rapid urease test. H. pylori infection was diagnosed according to the rapid urease test and histological appearance. The histological appearance of the gastric mucosa was evaluated according to modified Sydney criteria. A C-octanoic breath test was performed after a test meal (2 slices of toasted bread, 10 g margarine, and 1 egg with 100 microL of the tracer dipped in the yolk) with 13 points of air collection in 4 hours. RESULTS: Infection with H. pylori was observed in 12 of 27 patients (44.4%). The gastric emptying half-time was shorter in infected patients than in uninfected patients (mean +/- SD 153.4 +/- 20.0 min vs 179.2 +/- 32.2 min; P = 0.019), as was the lag phase (106.3 +/- 22.6 vs 126.6 +/- 22.7 min; P = 0.038). There was no relationship between gastric emptying (half-time and lag phase) and degree of histological abnormality. Vomiting and nausea were associated with slower gastric emptying in patients without H. pylori gastritis more often than in infected patients. CONCLUSIONS: The study suggests that delayed gastric emptying can play an important role in a subgroup of dyspeptic patients, particularly those without H. pylori infection who have nausea and vomiting.

[Analysis of clinicopathological, tumor cell proliferation and apoptosis parameters in adenocarcinoma of the gastric cardia]. / Estudo clínico-patológico, da proliferação celular e da apoptose no adenocarcinoma gástrico da cárdia.

BACKGROUND/AIMS: In view of the increased incidence of carcinoma of the cardia over recent years, this work had the aim of studying the clinicopathological aspects, cell proliferative and tumor apoptotic indices of this neoplasm, their interrelations and possible influences on the prognosis. MATERIAL AND METHODS: Forty cases of adenocarcinoma of the cardia were studied between 1988 and 2001, with a minimum clinical follow-up of 3 years. Patients were excluded if they had previous chemotherapy or radiotherapy treatment, presented early neoplasia, or died during the operations or for other reasons unrelated to cancer. Gender; age, Laurén and Ming histological type, staging, and the presence or absence of intestinal metaplasia, epithelial dysplasia and Helicobacter pylori in the adjacent mucosa were analyzed. The apoptotic index was evaluated via hematoxylin-eosin in the primary tumor. To analyze the cell proliferation tumor, PCNA was utilized. The immunohistochemical technique utilized was streptavidin-biotin-peroxidase. For the survival analysis, cases with distant metastasis upon diagnosis were excluded. For the statistical analysis, the Student t and Mann-Whitney tests, Kaplan-Meier curves and Cox regression model were utilized. RESULTS: The mean age was 61 years (median: 63). There was predominance of the male gender (72.5%), diffuse histological type (55%) and infiltrative histological type (72.5%), and the more advanced stages (III and IV: 67.5%). There was no association with intestinal metaplasia and/or H. pylori. No epithelial dysplasia was detected in adjacent mucosa in any of the cases. The mean apoptotic index was 7.05 in 10 high power fields and 11.40 in 500 cells (2.28%). The mean positivity to PCNA was 275.05 cells in ten high power fields and 409.33 in 500 cells (81.9%). There was a positive correlation between the cell proliferative and apoptotic indices. There was a positive correlation for intestinal histological type with PCNA and apoptotic indices, in 10 high power fields. The mean survival was 28.41 months. Age over 63 and apoptotic index over 7.05 showed a negative correlation with survival, in multivariate analysis. CONCLUSIONS: Adenocarcinoma of the cardia predominated in male adults of mean age 61 years, and the predominant type was diffuse in more advanced stages. There was a correlation between apoptosis and tumoral cell proliferation. Survival in cases of adenocarcinoma of the cardia is still low. Both age and apoptosis were independent prognostic factors in cancer of the cardia.

Aspectos funcionais, microbiológicos e morfológicos intestinais em crianças infectadas pelo vírus da imunodeficiência humana / Functional, microbiological and morphological intestinal findings among human immunodeficiency virus infected children

RACIONAL: O trato gastrointestinal é freqüentemente acometido nas crianças infectadas pelo vírus da imunodeficiência humana, com importantes repercussões no seu estado nutricional e sobrevida. A maioria dos estudos relacionados a esse tema foi desenvolvida com adultos, sendo menos investigado o problema nas crianças OBJETIVOS: Estudar aspectos digestivo-absortivos, microbiológicos e morfológicos intestinais em crianças infectadas pelo vírus da imunodeficiência humana MATERIAL E MÉTODOS: Onze crianças infectadas pelo vírus da imunodeficiência humana, menores de 13 anos, pertencentes às categorias clínicas A, B ou C, divididas em dois grupos: cinco pacientes com relato atual ou recente de diarréia e seis pacientes sem diarréia nos 30 dias que antecederam à inclusão no estudo. Investigação proposta: biopsia de intestino delgado e reto para análise morfológica e microbiológica, coprocultura, protoparasitológico de fezes, pesquisa de rotavírus, micobactérias e Cryptosporidium; teste da D-xilose RESULTADOS: Todos os pacientes testados (9/11) apresentavam má absorção da D-xilose (8,4-24,4 mg/dL). Os achados histopatológicos de intestino delgado foram inespecíficos, representados em sua maioria, por enteropatia grau I a II (6/10). Em todos os casos foi constatado aumento do infiltrado celular do córion. As alterações histopatológicas do reto também foram inespecíficas, com presença de aumento do infiltrado celular do córion. A pesquisa de microorganismos enteropatogênicos só foi positiva em dois casos, sendo identificado Mycobacterium avium intracellulare e Cryptosporidium nas fezes CONCLUSÕES: Demonstrou-se alta prevalência (100 por cento) de má absorção intestinal em crianças infectadas pelo vírus da imunodeficiência humana, com ou sem diarréia. Não foi possível estabelecer correlações quanto à presença de agentes enteropatogênicos, má absorção intestinal, alterações morfológicas intestinais e ocorrência ou não de diarréia. Não houve correlação...

BACKGROUD: Gastrointestinal tract disorders are frequent among human immunodeficiency virus infected children, with important repercussions on nutrition and survival. Most studies related to this subject were restricted to adults, being less investigated the problem in the children. AIMS: To study intestinal digestion, absorption, microbiological and morphological findings among human immunodeficiency virus infected children. MATERIAL AND METHODS: Eleven human immunodeficiency virus infected children under 13 years old, belonging to clinical categories A, B or C, separated in two groups: five patients with current or recent episode of diarrhea and six patients without diarrhea in the last 30 days preceding entering in study. Investigation proposed: microbiological and morphological analysis of small intestine and rectum biopsy; stool exams for bacterium, parasite, rotavirus, Mycobacterium species and Cryptosporidium; D-xylose test RESULTS: All tested subjects (9/11) had low D-xylose absorption (8,4 _ 24,4 mg d/L). Small intestinal mucosa histology findings were nonspecific, represented, in majority, of grade I/II enteropathy (6/10). Increased cellular infiltration of the chorion was observed in all specimens. Rectum histology alterations were also nonspecific, with chorion increased cellular infiltration. Mycobacterim avium intracellulare and Cryptosporidium were the solely microorganisms founded, both in stool CONCLUSIONS: Our study demonstrated high prevalence (100 percent) of intestinal malabsorption among human immunodeficiency virus infected children, despite the occurrence or not of diarrhea. It was not possible to establish relationships between the presence of microorganisms, intestinal malabsorption, intestinal morphologic findings and the occurrence or not of diarrhea. There was no correlation between D-xylose and intensity of villous atrophy.

Estudo clínico-patológico, da proliferação celular e da apoptose no adenocarcinoma gástrico da cárdia / Analysis of clinicopathological, tumor cell proliferation and apoptosis parameters in adenocarcinoma of the gastric cardia / Analysis of clinicopathological, tumor cell proliferation and apoptosis parameters in adenocarcinoma of the gastric cardia

RACIONAL/OBJETIVO: Em vista do aumento na incidência do carcinoma da cárdia nos últimos anos, este trabalho visa estudar os aspectos clínico-patológicos, da proliferação celular e da apoptose tumorais, suas correlações e eventuais influências no seu prognóstico. MATERIAL E MÉTODO: Estudaram-se 40 doentes submetidos a gastrectomia total por carcinoma da cárdia, no período de 1988 a 2001, com seguimento clínico de no mínimo 3 anos. Excluíram-se aqueles com tratamento químio ou radioterápico prévio, neoplasia precoce, óbito no intra-operatório ou por outras causas não relacionadas ao câncer. Analisou-se sexo, idade, tipo histológico de Laurén, padrão de crescimento tumoral de Ming, estádio e presença ou ausência de metaplasia intestinal e/ou Helicobacter pylori na mucosa adjacente. O índice apoptótico foi avaliado por cortes histológicos corados pela hematoxilina-eosina. O índice de proliferação celular foi avaliado por meio da imunoexpressão ao PCNA, sendo a técnica imunoistoquímica utilizada a da estreptoavidina-biotina-peroxidase. Para análise da sobrevida, excluíram-se os casos com metástase à distância ao diagnóstico. Foram utilizados testes t de Student, de Mann-Whitney, curvas de Kaplan-Meier e modelo de regressão de Cox. O nível de significância adotado foi menor de 0,05. RESULTADOS: A idade média foi de 61 anos (mediana: 63); houve predomínio do sexo masculino (72,5 por cento), tipo histológico difuso (55 por cento) e infiltrativo (72,5 por cento) e de estádios mais avançados (III e IV: 67,5 por cento). Não houve associação com metaplasia intestinal e/ou H. pylori. O índice apoptótico médio foi de 7,05 em 10 campos consecutivos de grande aumento e de 11,40 em 500 células (2,28 por cento). A positividade média ao PCNA foi de 275,05 células em 10 campos consecutivos de grande aumento e de 409,33 em 500 células (81,9 por cento). Houve correlação positiva do tipo histológico intestinal com PCNA e com a apoptose em 10 campos consecutivos de grande aumento e de ...

BACKGROUND/AIMS: In view of the increased incidence of carcinoma of the cardia over recent years, this work had the aim of studying the clinicopathological aspects, cell proliferative and tumor apoptotic indices of this neoplasm, their interrelations and possible influences on the prognosis. MATERIAL AND METHODS: Forty cases of adenocarcinoma of the cardia were studied between 1988 and 2001, with a minimum clinical follow-up of 3 years. Patients were excluded if they had previous chemotherapy or radiotherapy treatment, presented early neoplasia, or died during the operations or for other reasons unrelated to cancer. Gender; age, Laurén and Ming histological type, staging, and the presence or absence of intestinal metaplasia, epithelial dysplasia and Helicobacter pylori in the adjacent mucosa were analyzed. The apoptotic index was evaluated via hematoxylin-eosin in the primary tumor. To analyze the cell proliferation tumor, PCNA was utilized. The immunohistochemical technique utilized was streptavidin-biotin-peroxidase. For the survival analysis, cases with distant metastasis upon diagnosis were excluded. For the statistical analysis, the Student t and Mann-Whitney tests, Kaplan-Meier curves and Cox regression model were utilized. RESULTS: The mean age was 61 years (median: 63). There was predominance of the male gender (72.5 percent), diffuse histological type (55 percent) and infiltrative histological type (72.5 percent), and the more advanced stages (III and IV: 67.5 percent). There was no association with intestinal metaplasia and/or H. pylori. No epithelial dysplasia was detected in adjacent mucosa in any of the cases. The mean apoptotic index was 7.05 in 10 high power fields and 11.40 in 500 cells (2.28 percent). The mean positivity to PCNA was 275.05 cells in ten high power fields and 409.33 in 500 cells (81.9 percent). There was a positive correlation between the cell proliferative and apoptotic indices. There was a positive correlation for intestinal ...

Glycine reduces tissue lipid peroxidation in hypoxia-reoxygenation-induced necrotizing enterocolitis in rats.

PURPOSE: To assess the protective effect of glycine in an experimental model of Neonatal Necrotizing Enterocolitis (NEC). METHODS: Fifty (50) neonatal Wistar rats, from a litter of six female rats and weighing 4 to 6 grams, were used. Five animals were cannibalized and the 45 remaining were distributed into three groups: the G1 normal control group (n=12); the G2 Group (n=16), of animals that underwent hypoxia-reoxygenation (HR); the G3 Group of animals (n=17) that underwent HR following a 5% intraperitoneal glycine infusion. The animals underwent hypoxia in a CO2 chamber receiving an air flow of 100% CO2 for 5 minutes and reoxygenation receiving an O2 flow at 100% for 5 minutes. One centimeter long small bowel and colon segments were prepared for histological analysis. The rest of the bowel was removed in a block and frozen at minus 80 degrees C for homogenization and determination of tissue malondialdehyde (MDA). Tissue lesions were classified as Grade 0 to Grade 5, according to the level of damaged mucosa. RESULTS: The animals in Group G1 had levels of small bowel and colon lesion significantly smaller as compared to the animals in Groups G2 and G3. The G2 group had mean MDA values significantly higher than the animals in the G1 (p = .015) and G3 (p=0.021) groups. MDA values did not differ significantly (p = 0.992) for the animals in groups G1 and G3. CONCLUSION: Glycine reduces tissue MDA levels (a measurement of lipid peroxidation) following HR in neonatal rats.

Glycine reduces tissue lipid peroxidation in hypoxia-reoxygenation-induced necrotizing enterocolitis in rats

OBJETIVO: Avaliar o efeito protetor da glicina, num modelo experimental de ECN. MÉTODOS: Foram utilizados 50 ratos Wistar recém-nascidos, com peso variando de 4 a 6 gramas, provenientes da ninhada de seis ratas. Cinco animais foram canibalizados e, os 45 restantes, foram distribuídos em três grupos: controle G1(n=12); G2(n=16), animais que foram submetidos à hipóxia-reoxigenação; G3(n=17), animais submetidos à hipóxia-reoxigenação após uma infusão intraperitoneal de glicina 5%. Os animais foram submetidos à hipóxia em uma câmara de CO2 recebendo um fluxo de ar contendo 100% de CO2, durante 5 minutos e à reoxigenação recebendo um fluxo de O2 a 100% por 5 minutos. Segmentos de intestino delgado e cólon de 1 cm de extensão foram preparados para análise histológica. O restante do intestino foi removido em bloco e congelado a menos 80°C para homogeneização e dosagem de malondialdeído tecidual (MDA). Classificou-se as lesões teciduais de Grau 0 a Grau 5, de acordo com a extensão da lesão mucosa. RESULTADOS: Os animais do Grupo G1 apresentaram graus de lesão de intestino delgado e cólon significantemente menores do que os animais dos Grupos G2 e G3. O grupo G2 apresentou valores médios de MDA significantemente maiores do que os animais do grupo G1 (p = .015) e G3 (p=0.021). Os animais dos grupos G1 e G3 apresentaram valores de MDA que não diferiram de forma significante (p = 0.992). CONCLUSÃO: A glicina diminuiu os níveis de MDA intestinais (um marcador da peroxidação lipídica) em ratos neonatais submetidos à hipóxia-reoxigenação.

[Evaluation of an experimental model of necrotizing enterocolitis in rats]. / Avaliação de um modelo experimental de enterocolite necrosante neonatal em ratos.

OBJECTIVE: To evaluate an experimental model of necrotizing enterocolitis in rats proposed by OKUR e col. in 1995. METHODS: On their first day of life, 28 EPM-Wistar rats weighing between 4 and 6 grams were submitted to hypoxia (H) by placing them in a CO2 gas chamber for rodents' sacrifice, where they received a 100% CO2 air flow for 5 minutes. After the hypoxia the animals were reanimated (R) with a 100% O2 air flow, also for 5 minutes. The animals were allocated in two groups: G1: control (n=12): rats not submitted to H-R; G2: (n=16): rats submitted to H-R. Segments of the small intestine and colon were prepared for histological analysis. The remaining intestine was used to measure tissular malondialdehyde. RESULTS: Mean malondialdehyde dosages were 1.05 (0.44-2.03) and 2.60 (0.59- 6.4) nmol MDA/mg protein for G1 and G2, respectively. G2's mean value was significantly higher than in the control group (p<0.002). Significant statistical difference between the studied groups was found in relation the level of injury, with G1 presenting significantly lower levels than G2. CONCLUSIONS: The model showed that neonatal hypoxia may cause intestinal wall injury in rats. Despite the discreet histological injuries found, the method is suitable for evaluation of tissular free radicals.

Avaliação de um modelo experimental de enterocolite necrosante neonatal em ratos / Evaluation of an experimental model of necrotizing enterocolitis in rats

OBJETIVO: Avaliar um modelo experimental de enterocolite necrosante em ratos proposto por Okur e colaboradores em 1995. MÉTODOS: Utilizou-se 28 ratos da raça EPM-Wistar no primeiro dia de vida, com peso entre 4 a 6 gramas. Os animais foram submetidos a hipóxia (H) colocando os filhotes em uma câmara de gás CO2 para sacrifício de roedores onde receberam um fluxo de ar contendo 100 por cento de CO2 durante 5 minutos. Após a hipóxia os animais foram reanimados (R) com fluxo de ar contendo O2 a 100 por cento, também durante 5 minutos. Os animais divididos em dois grupos: G1: controle (n=12): ratos não submetidos a H-R; G2: (n=16): ratos submetidos a H-R. Segmentos de intestino delgado e cólon foram preparados para análise histológica. O restante do intestino foi utilizado para dosagem de malondialdeído tecidual. RESULTADOS: Dosagem de malondialdeído do G1 foi em média 1,05 (0,44-2,03) e do G2 foi em média 2,60 (0,59- 6,4) nmol MDA/mg proteína. O G2 teve média significativamente maior do que a do grupo controle (p<0,002). Foi encontrada diferença estatisticamente significante entre os grupos de estudo quanto à distribuição do grau de lesão onde o grupo G1 apresentou graus significantemente menores do que o grupo G2. CONCLUSÕES: O modelo mostrou que a hipóxia neonatal em ratos provoca lesões na parede intestinal.. Apesar das lesões histológicas discretas é um bom método para avaliação da liberação de radicais livres teciduais.

[Functional, microbiological and morphological intestinal findings among human immunodeficiency virus infected children]. / Aspectos funcionais, microbiológicos e morfológicos intestinais em crianças infectadas pelo vírus da imunodeficiência humana.

BACKGROUND: [corrected] Gastrointestinal tract disorders are frequent among human immunodeficiency virus infected children, with important repercussions on nutrition and survival. Most studies related to this subject were restricted to adults, being less investigated the problem in the children. AIMS: To study intestinal digestion, absorption, microbiological and morphological findings among human immunodeficiency virus infected children. MATERIAL AND METHODS: Eleven human immunodeficiency virus infected children under 13 years old, belonging to clinical categories A, B or C, separated in two groups: five patients with current or recent episode of diarrhea and six patients without diarrhea in the last 30 days preceding entering in study. Investigation proposed: microbiological and morphological analysis of small intestine and rectum biopsy; stool exams for bacterium, parasite, rotavirus, Mycobacterium species and Cryptosporidium; D-xylose test RESULTS: All tested subjects (9/11) had low D-xylose absorption (8,4 _ 24,4 mg d/L). Small intestinal mucosa histology findings were nonspecific, represented, in majority, of grade I/II enteropathy (6/10). Increased cellular infiltration of the chorion was observed in all specimens. Rectum histology alterations were also nonspecific, with chorion increased cellular infiltration. Mycobacterim avium intracellulare and Cryptosporidium were the solely microorganisms founded, both in stool CONCLUSIONS: Our study demonstrated high prevalence (100%) of intestinal malabsorption among human immunodeficiency virus infected children, despite the occurrence or not of diarrhea. It was not possible to establish relationships between the presence of microorganisms, intestinal malabsorption, intestinal morphologic findings and the occurrence or not of diarrhea. There was no correlation between D-xylose and intensity of villous atrophy.

Clinicopathologic study and Ki-67 proliferative marker evaluation in human osteosarcomas

BACKGROUND: Osteosarcoma is a relatively uncommon malignant neoplasm and little information has been reported on its cell proliferation using Ki-67. OBJECTIVES: Evaluate histological, clinical and immunohistochemical parameters using Ki-67 labeling index (LI), correlate one to another and with follow-up. MATERIAL AND METHOD: Fifty-seven patients with available clinicopathological data submitted to the study of cell proliferation as determined by Ki-67 expression measured by immunohistochemistry (IHC) staining using formalin-fixed paraffin embedded sections. In each sample, positive cells were quantified on at least a thousand nuclei and expressed as Ki-67 LI according to median value. RESULTS AND DISCUSSION: Non-significant correlations were observed in metastatic and non-metastatic cases when variables as surgery, tumor size, death and relapse were compared with Ki-67 LI values (cut-off of 45 percent). In the group of non-metastatic cases there was a direct correlation between higher values of Ki-67 index and better overall survival. Metastatic patients overall survival curve and LI high and low Ki-67 did not show significant differences. CONCLUSION: Based on our results the Ki-67 LI could be useful as a prognostic marker in patients without metastasis at diagnosis.

INTRODUÇÃO: Osteossarcoma é uma neoplasia rara do tecido ósseo, tendo sido publicados poucos trabalhos na literatura que avaliaram o índice de proliferação celular Ki-67 (IPC-Ki-67) nesse tipo de sarcoma. OBJETIVO: Avaliação e comparação dos parâmetros clínicos, histológicos e de imunoexpressão do Ki-67 com a evolução clínica dos pacientes. MATERIAL E MÉTODO: Blocos de parafina de 57 casos de osteossarcomas, cujos pacientes apresentavam dados clinicopatológicos fidedignos, foram submetidos a estudo da proliferação celular através da técnica de imuno-histoquímica. Em cada amostra o IPC foi determinado pela contagem de núcleos marcados em pelo menos mil células neoplásicas. Em cada grupo de pacientes o IPC-Ki-67 foi considerado alto ou baixo, tendo como referência a mediana dos valores do Ki-67. RESULTADOS E DISCUSSÃO: Não houve correlação estatística significativa entre os valores do IPC-Ki-67 (valor de corte = 45 por cento) dos osteossarcomas dos pacientes com ou sem metástases ao diagnóstico, em relação às variáveis óbito, recaída tumoral, tamanho do tumor e tipo de cirurgia. No grupo de pacientes sem metástases houve correlação direta entre o tempo de sobrevida e o índice de proliferação celular (isto é, o IPC-Ki-67 > 45 por cento correlacionou-se com maior tempo de sobrevida). Nos casos de pacientes com metástases ao diagnóstico, os valores do IPC-Ki-67 não apresentaram correlação significativa com a sobrevida. CONCLUSÃO: Com base nos nossos resultados, o IPC-Ki-67 pode ser utilizado para avaliação prognóstica em pacientes com osteossarcoma sem metástases ao diagnóstico.

Abordagem citogenética e molecular em material de abortos espontâneos / Cytogenetic and molecular evaluation of spontaneous abortion samples

OBJETIVOS: avaliar o desempenho da citogenética e das técnicas de hibridização in situ fluorescente (FISH) e reação em cadeia da polimerase (PCR) no estudo das aneuploidias cromossômicas numéricas e na determinação do sexo fetal em amostras de abortos espontâneos. MÉTODOS: duzentos e dezenove amostras de produtos de abortos espontâneos foram submetidas a estudo citogenético. Deste total, 40 amostras foram também submetidas à técnica de PCR-nested para a determinação do sexo fetal: 32 foram selecionadas devido à falha de crescimento no estudo citogenético e oito foram escolhidas ao acaso. Vinte amostras foram selecionadas para detecção de aneuploidias cromossômicas pela técnica de FISH, utilizando-se sondas para os cromossomos 13, 18, 21, X e Y: 13 casos foram submetidos a FISH devido à falha de crescimento no estudo citogenético e sete foram escolhidos ao acaso. Foi calculada a taxa de sucesso (obtenção de cariótipo) de cada técnica. Para comparação das taxas de sucesso foi utilizado o teste de chi2, sendo considerados significantes resultados com p<0,05. Foi avaliado o índice de acerto entre os resultados das amostras submetidas a mais de um exame, tomando-se como padrão-ouro o resultado do estudo citogenético. RESULTADOS: houve crescimento celular em 84,9 por cento das amostras submetidas a análise citogenética. Em 51,1 por cento dos casos foram encontradas alterações cromossômicas: 65,2 por cento trissomias, 17,9 por cento triploidias, 9,4 por cento tetraploidias, 4,2 por cento monossomia do cromossomo X e 1,1 por cento trissomia dupla, tetrassomia e alteração estrutural. A trissomia mais freqüente foi a do cromossomo 16 (39 por cento). FISH e PCR tiveram taxa de sucesso de 90 por cento, não diferindo significativamente do exame citogenético. Em todos os casos submetidos a mais de um exame os resultados foram concordantes. Nas amostras com falha de crescimento celular no exame citogenético e submetidas a outra técnica, a PCR obteve sucesso em 87,5 por cento e a FISH em 84,6 por cento. CONCLUSAO: o estudo citogenético de restos ovulares de abortamentos espontâneos teve elevada taxa de sucesso e evidenciou anomalias cromossômicas em mais da metade dos casos. As técnicas de biologia molecular (PCR-nested e FISH) complementaram o estudo citogenético e permitiram a obtenção de resultados seguros na detecção de alterações cromossômicas numéricas e na determinação do sexo fetal

13C-urea breath test to diagnose Helicobacter pylori infection in children aged up to 6 years.

BACKGROUND: 13C-urea breath test (13C-UBT) is an accurate noninvasive tool for diagnosis of Helicobacter pylori infection. It is considered the best method for epidemiological studies, but there are few studies to evaluate the 13C-UBT in infants and toddlers. AIM: To evaluate the 13C-UBT performed with infrared spectroscopy in children aged up to 6 years. PATIENTS: Sixty-eight patients (6 months. to 5 years 11 months.) were evaluated prospectively and consecutively. METHODS: Helicobacter pylori infection was detected by positive culture, or rapid urease test and histological examination, both positive. 13C-UBT was performed with 50 mg of 13C-urea diluted in 100 ml of commercial orange juice. Two expired air samples were collected: before and 30 minutes after tracer ingestion. Cutoff of delta over baseline (DOB) was 4.0 per thousand and urea hydrolysis rate 10 microg/minute. RESULTS: Fifteen of 68 (22.1%) patients were H. pylori infected. Sensitivity was 93.3% (95% CI; 86.8%-99.7%) and specificity was 96.2% (95% CI; 93.6%-98.8%), and these values were equal for DOB and urea hydrolysis rate. Negative DOB values in noninfected patients ranged from -1.5 per thousand to 2.6 per thousand and positive DOB values ranged from 10.8 per thousand to 105.5 per thousand. There was no relationship between DOB values and age. Conclusion. 13C-UBT performed with infrared spectroscopy proved to be a reliable and accurate noninvasive diagnostic tool for H. pylori infection detection in children aged up to 6 years. Results far from cutoff value can clearly distinguish positive from negative 13C-UBT results in children up to 6 years old.