Application of fluorescence in situ hybridization (FISH) as a tool to aid cytogenetics in 1,409 fetal samples.

AIM: To evaluate the technical application of fluorescence in situ hybridization (FISH) as a support to classical cytogenetic in numerical chromosomal aneuploidies studies in samples of amniotic fluid, chorionic villus, and fetal loss. MATERIALS AND METHODS: The authois performed cytogenetic analyses in 1,409 patients (678 amniocentesis, 512 chorionic villus samples, and 219 spontaneous abortions) during one year. FISH molecular study aided traditional cytogenetic in 90 cases. These cases were indicated based on the diagnostic hypothesis of each patient or when no cellular growth was obtained. The authors standardized the FISH in discoloured slides. RESULTS: They had 85% positive FISH in amniotic fluid, 70% in chorionic villus, and 90% in abortion material using 13, 18, 21 X and Y centromeric probes. It showed 12% of altered FISH in amniotic fluid (100% trisomies), 10% in chorionic'villus (50% trisomy and 50% X - monosomy), and 22% in abortion material (50% trisomy, 25% X-monosomy, and 25% triploidy). FISH and cytogenetic analysis confirmed the results. CONCLUSION: This technique revolutionized clinical and research applications of cytogenetics. In this particular paper, FISH was a valuable and reliable technique to promptly identify rapid detection of aneuploidies in interphase cells, metaphase spread and paraffin-embedded samples. It is hoped that, in the future, the economic viability of array CGH and FISH, with the decreasing cost of testing and their genomics advantages can be incorporated as routine and customized in the approach of prenatal diagnosis.

The regular arrangement of collecting venules pattern evaluated by standard endoscope and the absence of antrum nodularity are highly indicative of Helicobacter pylori uninfected gastric mucosa.

AIM: To evaluate the accuracy of antrum nodularity and the regular arrangement of collecting venules for diagnosing Helicobacter pylori gastritis. METHODS: Ninety-nine consecutive children and adolescents (1.07 years-17.69 years, mean+/-S.D.=9.71+/-3.80 F:M 54:45) undergoing upper digestive endoscopy were assessed for the presence of antrum nodularity and regular arrangement of collecting venules pattern to determine the status of H. pylori infection. Antrum nodularity was observed by a tangential view of the greater curvature of the gastric antrum. Regular arrangement of collecting venules was visualized as being the regular pattern of red points evaluated with a standard endoscope. Two biopsies from the antrum were collected for histology and rapid urease test. The accuracy of diagnosis based on antrum nodularity and regular arrangement of collecting venules was evaluated considering the sensitivity, specificity and likelihood ratio. RESULTS: H. pylori was detected in 32/99 patients (32.3%). Antrum nodularity provided 59.4% sensitivity (95% confidence interval 50.7-68.1), 98.5% specificity (95% confidence interval: 97-100), likelihood ratio+ 39.78, and likelihood ratio- 0.41. A regular arrangement of collecting venules pattern provided 96.9% sensitivity (95% confidence interval: 93.8-100), 88.1% specificity (95% confidence interval: 84.1-92), likelihood ratio+ 8.11, and likelihood ratio- 0.04. CONCLUSION: Antrum nodularity is a specific finding, although its sensitivity is low. A regular arrangement of collecting venules pattern and the absence of antrum nodularity are highly indicative of normal gastric mucosa that is negative for Helicobacter pylori.

Nitric oxide levels in the intestines of mice submitted to ischemia and reperfusion: L-arginine effects.

OBJECTIVE: Usually an experimental necrotizing enterocolitis experimental model, we Investigated nitric oxide levels in intestinal tissues of newborn mice with or without l-arginine therapy during sessions of ischemia and reoxygenation. METHODS: Twenty-six newborn mice from the Wistar EPM-1 lineage, weighing from 4.5 to 6.2 g, were randomly assigned to three groups: G-I/R, hypoxia and reoxygenation; G-Arg, l-arginine treatment I/R; and G-CTL, controls. G-I/R and G-Arg mice underwent twice a day during their first 3 days of life exposure to gas chambers with 100% CO(2) for 5 minutes at 22 degrees C before reoxygenation with 100% O(2) for another 5 minutes. After 12 hours, all animals were sedated, laparotomized, and had samples of ileum and colon taken and- either formalin fixed histopathologic examinations or frozen to -80 degrees C for estimation of tissue nitric oxide levels. Intestinal injuries were classified according to the criteria of Chiu et al. RESULTS: The G-I/R and G-Arg groups showed injuries characteristic of necrotizing enterocolitis (NEC) with an improved structural preservation rate in G-Arg. The concentration of nitric oxide in the Ileum was much higher with G-Arg (16.5 +/- 4.9; P = 0.0019) G-I/R (7.3 +/- 2.0). This effect was not observed in the colon: G-I/R = 10.7 +/- 4.6 versus G-Arg = 15.5 +/- 8.7 (P = .2480). CONCLUSION: Supply of L-arginine increased tissue levels of nitricoxide and reduced morphologic intestinal injury among mice undergoing I/R.

Carditis is related to Helicobacter pylori infection in dyspeptic children and adolescents.

BACKGROUND: Etiology of gastric cardia inflammation is still controversial. AIMS: To evaluate the association between carditis and Helicobacter pylori infection and the correlation among inflammatory changes observed in biopsies taken from cardia, corpus, and antrum in a well-defined group of patients. PATIENTS: The mean age of 45 dyspeptic patients was 10.4 years (range 5.1-17.0 years); gender F/M rate: 1.6/1. METHODS: A total of 450 specimens from esophagus (2), cardia (2), corpus (3), and antrum (4) were collected for biopsy. The presence of H. pylori was assessed by histology and a rapid urease test. The types of glandular epithelium of cardia found in specimens were identified and both inflammatory changes and H. pylori density were graded. RESULTS: Carditis was present in specimens of 30/45 (66.7%) of the patients. Presence of H. pylori in specimens was detected in the antrum (26/45; 57.8%), in the corpus (19/45; 42.2%), and in the cardia (14/45; 31.1%). There was a strong association between carditis and presence of H. pylori infection (OR=27.08) by multivariate analysis. The scores for inflammation and activity in the cardia, corpus and antrum have shown a relationship except for both cardia and antrum H. pylori density and corpus and cardia activity. The intensity of gastritis and degree of colonization with H. pylori were significantly higher in the antrum than in both the corpus and the cardia. Pangastritis was highly associated to H. pylori infection in 22/25 (88%) of the patients. CONCLUSIONS: 1. Carditis is associated to H. pylori infection in children with symptoms of dyspepsia; 2. The degrees of gastritis found at the cardia were correlated to those at the antrum and body except for both cardia and antrum H. pylori density and corpus and cardia activity.

Repercussions of extensive small bowel resections in growing rats.

OBJECTIVE: To study the effects of extensive intestinal resection on growing rats, with regard to animal weight and histomorphometry of the remaining small intestine. METHODS: Forty growing rats were allocated according to the extent of small intestine resection: 60%, 70%, 80%, or 90%. The animals were weighed every week and observed for 30 days. Following sacrifice the remaining small intestine was resected, fixed in 10% formol for 24 hours, embedded in paraffin, and stained using hematoxylin and eosin. The histological changes in the remaining small intestine were assessed for the length and thickness of villi, the thickness of the muscle layer, and the number of intestinal glands. RESULTS: All growing rats showed a fall in body weight, although it was more significant with the largest intestinal resection (80% and 90%). Villus length and muscle thickness increased after 30 postoperative days in all rats, but the number of intestinal glands remained unaltered. CONCLUSION: Growing rats with greatest resection of small intestine (80% and 90%) had better intestinal adaptation and slower recovery of body weight.

Morfometria do intestino delgado em crianças com diarréia por Escherichia coli enteropatogênica clássica e enteropatia ambiental assintomática / Morphometrics of the small bowel in children with diarrhea due to classical enteropathogenic Escherichia coli and to environmental asymptomatic enteropathy

Infecçäo entérica por Escherichia coli enteropatogênica clássica (EPEC) pode causar diferentes graus de alteraçöes das vilosidades do intestino delgado.OBJETIVOS. Este estudo teve por objetivo: 1) avaliar as alteraçöes morfológicas da mucosa intestinal na diarréia aguda por EPEC, por meio da morfometria linear, e compará-la a um grupo controle; 2) comparar o número de LIE encontrado na diarréia aguda e/ou persistente por EPEC com aqueles encontrados no grupo controle; 3) pesquisar a presença de E. coli aderida à mucosa do intestinodelgado naquelas crianças que apresentaram diarréia com coprocultura positiva para EPEC. PACIENTES E MÉTODOS. Foram analisados 30 biópsias da mucosa do intestino delgado de crianças com diarréia aguda e/ou persistente, com coprocultura positiva para EPEC e 16 biópsias obtidas da mucosa do intestino delgado de crianças portadoras de enteropatia ambiental assintomática, que constituíram nosso grupo controle. Foram realizadas as seguintes análises morfométricas: RESULTADOS: Espessura total mucosa µ EPEC=279,6 EAA=445,1 p<0,001; Altura vilosidade µ EPEC 134,3 EAA 248,0 p<0,001; Extensäo da zona críptica µ EPEC 145,2 EAA 197,1 p<0,02; linfócitos intra-epiteliais EPEC 11,6 EAA 15,5 p<0,005. CONCLUSÄO. Presença de bactéria gram-negativa, portanto, provavelmente, EPEC, foi constatada em três dos 30 pacientes com diarréia, apresentando coprocultura positiva para EPEC. As alteraçöes morfométricas ocorrem, principalmente, às custas da diminuiçäo das visolidades intestinais.

Doença celíaca e enteropatia ambiental: diagnósticos diferencial em crianças de baixo nível sócio-econômico / Celiac diseases and environmental bowel diseases: differential diagnosis in children of low socioeconomic status

A alta prevalência de Enteropatia Ambiental em nosso meio e em determinados casos, sua semelhança clínica, funcional e histológica com Doença Celíca, näo permite o diagnóstico diferencial entre essas duas patologias. Foram estudadas 12 crianças portadoras de Enteropatia Ambiental, com diarréia crônica, alteraçäo da prova de absorçäo da D-Xilose e atrofia vilositária subtotal da mucosa do intestino delgado. O acompanhamento das crianças com dieta isenta de glúten só permitiu o diagnóstico diferencial em metade dos pacientes. Conclui-se ser necessário o desenvolvimento de técnicas auxiliares para o diagnóstico de Doença Celíaca

Symptomatic environmental enteropathy in infancy: bacterial proliferation, funcional and morphological alterations of the small bowel

Asymptomatic Environmental enteropathy may occur in the low social economic stratum of the population temporarily, thus contributing to marginal malnutrition. Bacterial proliferation in the upper portions of the small bowel may be responsible for alterations in the digestive-absorptive function leading to nutritional dwarfism. Forty infants of the Cidade Leonor slum who were under one year of age and presented with asymptomatic environmental enteropathy were investigated for digestive-absorptive function, bacterial proliferation in the small bowel lumen, and jejunal morphology. The mean value of the D-xylose absorption test was 21.0 ñ 10.0 mg per cent, significantly lower than controls (48.0 ñ 13.8 mg per cent) (p < 0.001). Bacterial proliferation in the small bowel of the colonic type was identified in 25 (62,5 per cent) of the infants and jejunal mucosa abnormalities were found in 29(72.5 per cent) of the specimens. Grade II villous atrophy was the most frequent alteration found. The inflammatory infiltration in the lamina propria increased in the total group, varying from light to severe. Alterations in the microecology of the small intestine can occur even in the absence of diarrhea. Therefore, the nonexistence of symptoms does not necessarily imply a condition of weillbeing. If true measures are proposed to improve the quality of life in these poor communities the whole infant population must be taken into account and not only the group with overt symptoms