Ex vivo model of rabbit intestinal epithelium applied to the study of colonization by enteroaggregative escherichia coli / Modelo ex vivo de epitélio intestinal de coelho aplicado ao estudo de colonização por Escherichia coli enteroagregativa

ABSTRACT BACKGROUND The diarrheal syndrome is considered a serious public health problem all over the world and is considered a major cause of morbidity and mortality in developing countries. The high incidence of enteroaggregative Escherichia coli in diarrheal syndromes classified as an emerging pathogen of gastrointestinal infections. After decades of study, your pathogenesis remains uncertain and has been investigated mainly using in vitro models of adhesion in cellular lines. OBJECTIVE The present study investigated the interaction of enteroaggregative Escherichia coli strains isolated from childhood diarrhea with rabbit ileal and colonic mucosa ex vivo, using the in vitro organ culture model. METHODS The in vitro adhesion assays using cultured tissue were performed with the strains co-incubated with intestinal fragments of ileum and colon over a period of 6 hours. Each strain was tested with three intestinal fragments for each region. The fragments were analysed by scanning electron microscopy. RESULTS Through scanning electron microscopy we observed that all strains adhered to rabbit ileal and colonic mucosa, with the typical aggregative adherence pattern of “stacked bricks” on the epithelium. However, the highest degree of adherence was observed on colonic mucosa. Threadlike structures were found in greater numbers in the ileum compared to the colon. CONCLUSION These data showed that enteroaggregative Escherichia coli may have a high tropism for the human colon, which was ratified by the higher degree of adherence on the rabbit colonic mucosa. Finally, data indicated that in vitro organ culture of intestinal mucosa from rabbit may be used to elucidate the enteroaggregative Escherichia coli pathogenesis.

RESUMO CONTEXTO A síndrome diarréica é considerada um grave problema de saúde pública em todo o mundo e é considerada uma das principais causas de morbidade e mortalidade nos países em desenvolvimento. A elevada incidência de Escherichia coli enteroagregativa nas síndromes diarreicas a classificou como um patógeno emergente de infecções gastrintestinais. Depois de décadas de estudo, sua patogênese ainda é incerta e tem sido investigada usando principalmente modelos in vitro de adesão em linhagens celulares. OBJETIVO O presente estudo investigou a interação de cepas de Escherichia coli enteroagregativa isoladas de diarreia infantil com mucosa ileal e colônica de coelho ex vivo, utilizando o modelo de cultura de órgão in vitro. MÉTODOS Os ensaios de adesão in vitro utilizando tecido cultivado foram realizados com as cepas co-incubadas com fragmentos intestinais de íleo e de cólon durante um período de 6 horas. Cada cepa foi testada em três fragmentos intestinais para cada região. Os fragmentos foram analisados por microscopia eletrônica de varredura. RESULTADOS Através da microscopia eletrônica de varredura observamos que todas as cepas aderiram a mucosa ileal e colônica de coelho, com o padrão de aderência agregativo típico de “tijolos empilhados” no epitélio. Entretanto, o maior grau de adesão foi observado na mucosa do cólon. Estruturas filiformes foram encontradas em maior número no íleo em comparação com o cólon. CONCLUSÃO Esses dados mostraram que Escherichia coli enteroagregativa pode ter um maior tropismo para o cólon humano, o que foi ratificado pelo maior grau de aderência na mucosa do cólon de coelho. Finalmente, os dados indicaram que a cultura de órgão in vitro da mucosa intestinal de coelho pode ser utilizado para elucidar a patogênese de Escherichia coli enteroagregativa.

Modulation of small intestinal homeostasis along with its microflora during acclimatization at simulated hypobaric hypoxia.

At high altitude (HA) hypobaric hypoxic environment manifested several pathophysiological consequences of which gastrointestinal (GI) disorder are very common phenomena. To explore the most possible clue behind this disorder intestinal flora, the major player of the GI functions, were subjected following simulated hypobaric hypoxic treatment in model animal. For this, male albino rats were exposed to 55 kPa (~ 4872.9 m) air pressure consecutively for 30 days for 8 h/day and its small intestinal microflora, their secreted digestive enzymes and stress induced marker protein were investigated of the luminal epithelia. It was observed that population density of total aerobes significantly decreased, but the quantity of total anaerobes and Escherichia coli increased significantly after 30 days of hypoxic stress. The population density of strict anaerobes like Bifidobacterium sp., Bacteroides sp. and Lactobacillus sp. and obligate anaerobes like Clostridium perfringens and Peptostreptococcus sp. were expanded along with their positive growth direction index (GDI). In relation to the huge multiplication of anaerobes the amount of gas formation as well as content of IgA and IgG increased in duration dependent manner. The activity of some luminal enzymes from microbial origin like α-amylase, gluco-amylase, proteinase, alkaline phosphatase and β-glucuronidase were also elevated in hypoxic condition. Besides, hypoxia induced in formation of malondialdehyde along with significant attenuation of catalase, glutathione peroxidase, superoxide dismutase activity and lowered GSH/GSSG pool in the intestinal epithelia. Histological study revealed disruption of intestinal epithelial barrier with higher infiltration of lymphocytes in lamina propia and atrophic structure. It can be concluded that hypoxia at HA modified GI microbial imprint and subsequently causes epithelial barrier dysfunction which may relate to the small intestinal dysfunction at HA.

C6, a new monoclonal antibody, reacts with the follicle-associated epithelium of calf ileal Peyer's patches

The follicle-associated epithelium (FAE) of Peyer's patches (PPs) contains M cells that are important for reducing mucosal immune responses by transporting antigens into the underlying lymphoid tissue. We generated a monoclonal antibody (C6) that reacted with the FAE of calf ileal PPs, and analyzed the characteristics of C6 using immunohistochemistry and Western blotting. FAE of the ileal PP was stained with C6 during both late fetal developmental and postnatal stages. Neither the villous epithelial cell nor intestinal crypt basal cells were stained at any developmental stage. During the prenatal stages, FAE of the jejunal PP was C6-negative. However, a few C6-positive cells were distributed diffusely in some FAE of the jejunal PPs during the postnatal stages. The protein molecular weight of the antigen recognized by C6 was approximately 45 kDa. These data show that C6 is useful for identifying the FAE in ileal PPs and further suggest that differentiation of the FAE in these areas is independent of external antigens.

C6, a new monoclonal antibody, reacts with the follicle-associated epithelium of calf ileal Peyer's patches

The follicle-associated epithelium (FAE) of Peyer's patches (PPs) contains M cells that are important for reducing mucosal immune responses by transporting antigens into the underlying lymphoid tissue. We generated a monoclonal antibody (C6) that reacted with the FAE of calf ileal PPs, and analyzed the characteristics of C6 using immunohistochemistry and Western blotting. FAE of the ileal PP was stained with C6 during both late fetal developmental and postnatal stages. Neither the villous epithelial cell nor intestinal crypt basal cells were stained at any developmental stage. During the prenatal stages, FAE of the jejunal PP was C6-negative. However, a few C6-positive cells were distributed diffusely in some FAE of the jejunal PPs during the postnatal stages. The protein molecular weight of the antigen recognized by C6 was approximately 45 kDa. These data show that C6 is useful for identifying the FAE in ileal PPs and further suggest that differentiation of the FAE in these areas is independent of external antigens.

Distribution of serotonin-immunoreactive enterochromaffin cells in the gastrointestinal tract of the least shrew (Cryptotis parva) / Distribución de las células enterocromafines serotonina-inmunorreactiva del tracto gastrointestinal de la musaraña enana (Cryptotis parva)

Serotonin is an important neurotransmitter in the central (CNS) and peripheral (PNS) nervous systems. It is involved in a variety of physiological processes both in the gut and in the CNS. The present study examined the distribution of serotonin containing enterochromaffin cells in the gastrointestinal tract (GIT) of a vomit competent species, the least shrew. These cells were easily recognized by their globular granules stained with the H&E and serotonin immune-positive stain. The immunoreactive enterochromaffin cells (IERCs) were mainly confined to the basal portion of the glandular epithelium and were distributed throughout the shrew stomach, small and large intestine. None was found to be associated with the mucosal epithelial lining. Moreover, their distribution and count varied in different regions of the GIT suggesting specific functions for these regions. The highest concentration of IERCs was found in the colon followed by the Jejunum. Appreciable numbers of IERCs were found in the stomach especially at the esophageo-gastric junction. The gastric location of the IERCs was mainly in the basal portion of the gland. However, some IERCs were associated with the parietal cells of the stomach. Two types of IERCs were observed: One with globular secretory granules in their apical portion of the cytoplasm which were located within the glandular epithelial cells facing the glandular lumen which release their secretions into the lumen; and the second were basally located, facing the lamina propria of the mucosa. Their secretory granules were not distinct in shape, and are most probably paracrine in their mode of secretions...

La serotonina es un importante neurotransmisor del sistema nervioso central (SNC) y periférico (SNP). Está implicado en una variedad de procesos fisiológicos, tanto en el intestino y el SNC. El presente estudio examinó la distribución de la serotonina contenida en las células enterocromafines del tracto gastrointestinal (TGI) de una especie competente al vómito, la musaraña enana. Estas células se reconocen fácilmente por sus gránulos globulares teñidas con H-E y la inmuno-tinción positiva para serotonina. Las células enterocromafines inmunorreactivas (CEI) se limitan principalmente a la parte basal del epitelio glandular y se distribuyeron por todo el estómago, intestino delgado e intestino grueso de la musaraña. Ninguna célula se encontró asociada al revestimiento epitelial mucoso. Además, su distribución y el recuento varió en diferentes regiones del TGI sugiriendo funciones específicas de estas regiones. La mayor concentración de CEI se encuentran en el colon seguido por el yeyuno. Números apreciables de CEI se encontraron en el estómago, especialmente en la unión esofago-gástrica. La ubicación de las CEI gástricas fue principalmente en la porción basal de la glándula. Sin embargo, algunas CEI se asociaron con las células parietales del estómago. Dos tipos de CEI se observaron, una con gránulos secretores globulares en su porción apical del citoplasma que se encuentra dentro de las células epiteliales glandulares que enfrenta el lumen glandular que liberan sus secreciones en el lumen, y el segundo se encuentra basalmente, frente a la lámina propia de la mucosa. Sus gránulos secretores no fueron diferentes en forma, y probablemente son más paracrinas en su modo de secreción...

Adherence of enteroaggregative Escherichia coli to the ileal and colonic mucosa: an in vitro study utilizing the scanning electron microscopy / Aderência da Escherichia coli enteroagregativa a mucosas ileal e colônica: estudo in vitro utilizando microscopia eletrônica de varredura

CONTEXT: Enteroaggregative Escherichia coli strains have been associated with persistent diarrhea in several developing countries. In vivo procedures with animal models, in vitro assays with cellular lines and in vitro organ culture with intestinal fragments have been utilized to study these bacteria and their pathogenicity. OBJECTIVE: The present experimental research assessed the pathogenic interactions of three enteroaggregative Escherichia coli strains, using the in vitro organ culture, in order to show the adherence to different regions of both, the ileal and the colonic mucosa and demonstrate possible mechanisms that could have the participation in the prolongation of diarrheiogenic process. METHODS: This study used intestinal fragments from terminal ileum and colon that were excised from pediatric patients undergoing intestinal surgeries and from adult patients that underwent to colonoscopic procedures. Each strain was tested with three intestinal fragments for each region. Tissue was fixed for scanning electron microscopic analysis. RESULTS: These bacteria colonized ileal and colonic mucosa in the typical stacked-brick configuration in the ileum and colon. In both regions, the strains were seen over a great amount of mucus and sometimes over the intact epithelium. In some regions, there is a probable evidence of effacement of the microvilli. It was possible to see adhered to the intestinal surface, bacteria fimbrial structures that could be responsible for the adherence process. CONCLUSION: In order to cause diarrhea, enteroaggregative Escherichia coli strains adhere to the intestinal mucosa, create a mucoid biofilm on the small bowel surface that could justify the digestive-absorptive abnormalities and consequently, prolonging the diarrhea.

CONTEXTO: Cepas de Escherichia coli enteroagregativa têm sido associadas à diarreia persistente em vários países em desenvolvimento. Procedimentos in vivo com modelos animais, cultura de órgão in vitro com fragmentos intestinais e ensaios in vitro com linhas celulares têm sido utilizados para estudar essas bactérias e a sua patogenicidade. OBJETIVO: A presente investigação experimental avaliou as interações patogênicas de três cepas de Escherichia coli enteroagregativa, usando cultura de órgão in vitro, para mostrar a aderência a diferentes regiões do intestino: íleo e cólons e demonstrar possíveis mecanismos que poderiam ter participação na perpetuação do processo diarréico. MÉTODOS: Este estudo usou fragmentos de íleo terminal e cólon que foram retirados de pacientes pediátricos submetidos a cirurgias intestinais e de pacientes adultos que foram submetidos a colonoscopias. Cada cepa foi testada com três fragmentos intestinais para cada região. O tecido foi fixado para análise sob microscopia eletrônica de varredura. RESULTADOS: Estas bactérias colonizaram mucosa ileal e colônica na configuração típica de pilhas de tijolos. Em ambas as regiões, as bactérias foram vistas sobre grande quantidade de muco e, às vezes, sobre o epitélio intacto. Em algumas áreas, há evidência de provável achatamento de vilosidades. Foi possível ver sobre a superfície intestinal, estruturas fimbriais bacterianas que poderiam estar relacionadas ao processo de adesão. CONCLUSÕES: Para causar diarreia, cepas de Escherichia coli enteroagregativa aderem à mucosa intestinal e criam um biofilme de muco sobre a superfície do intestino delgado, o que poderia justificar as anormalidades digestivo-absortivas e, por conseguinte, prolongar a diarreia.

Morphoquantitative study of the submucous plexus (of Meissner) of the jejunum-ileum of young and old guinea pigs / Estudo morfoquantitativo do plexo submucoso (de Meissner) do jejuno-íleo de cobaias jovens e velhas

OBJECTIVE: To study the aging of submucous plexus of the small intestine (jejunum-ileum) of the guinea pigs from the quantitative, structural and ultrastructural perspective. METHOD: Chemical preparations of membrane of the jejunum-ileum of old and young animals with the use of light and electronic microscope. RESULTS: The ganglia of young animals presented between 1 and 56 neurons and the old animals presented from 1 to 30 neurons. The mean density of the ganglia by cm² in the young jejunum-ileum was of 551±36.89 and in the old one 413±11.86. The density of the neurons was 5011±291.11 neurons/cm² average in young animals and 2918±120.70 neurons/cm² in the old ones. The size of the neurons varied in both age groups. The collagen fibers in the ganglia of old animals they were condensed. Degenerated mitochondrias in the interior of the cell were frequent in the old animals. CONCLUSION: In submucous plexus of the jejunum-ileum there is a loss of 38 percent of the neurons with aging.

OBJETIVO: Estudar o envelhecimento do plexo submucoso do intestino delgado (jejuno-íleo) das cobaias do ponto de vista quantitativo, estrutural e ultra-estrutural. MÉTODO: Preparados de membrana do jejuno-íleo de animais jovens e velhos com a utilização de microscopia de luz e eletrônica. RESULTADOS: Os gânglios de animais jovens apresentaram entre 1 e 56 neurônios e os animais velhos apresentaram de 1 a 30 neurônios. A densidade média dos gânglios por cm² no jejuno-íleo jovem foi de 551±36,89 e no velho foi de 413±11,86. A densidade dos neurônios foi de 5011±291,11 neurônios/cm² em média nos animais jovens e 2918±120,70 neurônios/cm² nos velhos. O tamanho dos neurônios variou em ambos os grupos etários. As fibras colágenas nos gânglios de animais velhos estavam mais condensadas. Mitocôndrias degeneradas no interior da célula foram freqüentes nos animais velhos. CONCLUSÃO: No plexo submucoso do jejuno-íleo há uma perda de 38 por cento dos neurônios com o envelhecimento.

Pathophysiology of enteroaggregative Escherichia coli infection: an experimental model utilizing transmission electron microscopy / Fisiopatologia da infecção pela Escherichia coli enteroagregativa: um modelo experimental utilizando microscopia eletrônica de transmissão

CONTEXT: Enteroaggregative Escherichia coli strains have been associated with persistent diarrhea in several developing countries. In vivo procedures with animal models as rat, rabbit and gnotobiotic piglets intestinal loops, in vitro assays with cellular lines like T84, Caco 2, HT29, HeLa e HEp-2 and in vitro organ culture with intestinal fragments have been applied to study these bacteria and their pathogenicity. OBJECTIVES: The present experimental research assessed the pathogenic interactions of three enteroaggregative Escherichia coli strains, using the in vitro organ culture, in order to observe and compare alterations in different regions of both, the ileal and the colonic mucosa. METHODS: This study applied intestinal fragments from terminal ileum and colon that were excised from pediatric and adult patients that underwent colonoscopic procedures. Tissue was fixed for transmission electron microscopic study. Each bacterium was tested with three intestinal fragments for each region. RESULTS: Enteroaggregative Escherichia coli strains colonized and provoked citotoxic effects in the ileal and colonic mucosa. Total or partial villi destruction, vacuolization of basal cytoplasm of the enterocytes, epithelium detachment, derangement of the structure and epithelial cell extrusion in ileal mucosa could explain the perpetuation of the diarrhea. Bacterial aggregates were seen in intestinal lumen associated with mucus and cellular debris and in the intercellular spaces of the destroyed epithelium, suggesting bacterial invasion that seemed to be secondary to the destruction of the tissue. CONCLUSIONS: Pathogenesis of persistent diarrhea should include alterations in the small bowel structures where the digestive-absorptive functions take place. In the colonic mucosa the inflammatory lesions could explain the occurrence of colitis.

CONTEXTO: A Escherichia coli enteroagregativa está associada à diarréia persistente em vários países em desenvolvimento. Procedimentos in vivo empregando modelos animais como ratos, coelhos e alças intestinais de suínos gnotobióticos, e modelos in vitro com linhas celulares, tais como: T84, Caco 2, HT29, HeLa e HEp-2 e cultura de órgão in vitro são empregados no estudo desta bactéria e de sua patogenicidade. OBJETIVOS: Neste trabalho foram avaliadas as interações de três cepas de Escherichia coli enteroagregativa usando cultura de órgão in vitro, com o objetivo de observar e comparar as alterações em diferentes regiões do intestino: mucosa ileal e mucosa colônica. MÉTODOS: Este estudo empregou fragmentos de íleo terminal e cólon extraídos de pacientes submetidos a colonoscopia. Os fragmentos intestinais infectados in vitro foram fixados para avaliação em microscopia eletrônica de transmissão. Cada cepa bacteriana foi testada com três fragmentos intestinais de cada região. RESULTADOS: As cepas estudadas colonizaram e provocaram efeitos citotóxicos no íleo e no cólon. Alterações na mucosa ileal, tais como: destruição parcial ou total das vilosidades, vacuolização do citoplasma basal dos enterócitos, destacamento do epitélio e desarranjo da estrutura com extrusão de células epiteliais poderiam explicar a perpetuação do processo diarréico. Agregados bacterianos foram vistos no lúmen intestinal associados a muco e restos celulares e nos espaços intercelulares do epitélio destruído sugerindo invasão bacteriana que pareceu ser secundária à destruição do tecido. CONCLUSÃO: A patogênese da diarréia persistente deve incluir alterações no intestino delgado aonde ocorrem as funções digestivo-absortivas. Na mucosa colônica as lesões inflamatórias observadas justificariam a ocorrência de colite.

Histochemical and ultrastructural effects of phytic acid on the mucosa of rat ileum treated with meloxicam

Non-steroidal anti-inflammatory drugs [NSAIDs] cause gastrointestinal damage both in the upper and lower gastrointestinal tract. New anti-inflammatory drugs have been developed in an attempt to improve their gastrointestinal side-effect profile. In spite of the numerous therapeutic advantages of meloxicam, it has, however, damaging effects on the gastrointestinal tract. Inositol hexaphosphate [IP6], also known as phytic acid, is a naturally occurring polyphosphorylated carbohydrate that is particularly abundant in cereals and legumes. It has antioxidant, antimitotic, hypocholesterolemic and hypolipidemic effects. The present study was carried out to evaluate the effect of phytic acid administration on the injurious effects of meloxicam on the mucosa of rat ileum. Thirty albino rats were divided into 3 groups; the first group [control] was put on a normal diet for 4 weeks; the second group received a diet containing 200 mg/kg meloxicam per day for 4 weeks; the third group received a diet containing 200 mg/kg meloxicam per day followed by orally intubated IP6 three times/week for 4 weeks, at a total dose of 40 mg/kg. Slices of ileum of all groups were taken and subjected to histological, histochemical and ultrastructural investigation. The results confirmed the damaging effects of meloxicam on the ileum mucosa and revealed the ameliorative effects of phytic acid against these lesions

A comparative study of the ileum in rat (Rattus norvegicus), Bat (Eidolon helvum) and Pangolin (Manis tricuspis) as investigated using histological method / Estudio comparativo del íleon en rata (Rattus norvegicus), murciélago (Eidolon helvum) y pangolín (Manis tricuspis) utilizando como investigación el método histológico

The histological and morphometric differences in some parts of the gastrointestinal tracts of rat, bat and pangolin have been well established. This investigation aims at comparatively elucidating any adaptational changes the ileum of the three mammals must have adopted to meet its dietary requirements and also cope with their morphological differences. The investigation was carried out using ten rats, ten bats and ten pangolins of both sexes. The animals were slightly anaesthetized under chloroform inhalation. The intestines were harvested and the ileum excised and fixed in 10 percent formol saline. The tissues were processed for light microscopic study. The following stains were employed: Haematoxylin and eosin (H & E), Van Gieson and Verhoeff's haematoxylin elastic tissue stain. Stained slides were also analysed morphometrically. The results revealed microstructural modifications which characterized the mucosa as well as the pattern of distribution of the conjuntive tissue fibers in the ileum of the three mammals. These modifications are means of successfully coping with their respective diets and morphological differences.

Las diferencias histológicas y morfométricas en algunas partes del tubo digestivo de ratas, murciélagos y pangolines han sido bien establecidos. Esta investigación tiene por objeto aclarar comparativamente cualquier cambio de adaptación del íleon de los tres mamíferos que hayan adoptado para satisfacer sus necesidades dietéticas y también hacer frente a sus diferencias morfológicas. La investigación se llevó a cabo usando 10 ratas, 10 murciélagos y 10 pangolines, de ambos sexos. Los animales fueron ligeramente anestesiados bajo inhalación de cloroformo. Los intestinos fueron expuestos, el íleon extirpado y fijado en formol salino al 10 por ciento. Los tejidos fueron procesados para su estudio por microscopía de luz. Los siguientes tinciones fueron empleadas: Hematoxilina y eosina (H & E), Van Gieson y tinción de hematoxilina para tejidos elásticos de Verhoeff. Los cortes teñidos también se analizaron morfométricamente. Los resultados revelaron modificaciones microestructurales características de la mucosa, así como el patrón de distribución de las fibras de tejido conjuntivo en el íleon de los tres mamíferos. Estas modificaciones son el medio para entender sus diferencias en la dieta y morfología respectivamente.

Experimental colitis is associated with ultrastructural changes in inflamed and uninflamed regions of the gastrointestinal tract

The objective of this study was to examine theultrastructural changes in cell organelles such as mitochondria, endoplasmic reticulum [ER] and Golgi apparatus in inflamed colon and uninflamed ileum in colitic rats. Colitis was induced in rats by intracolonic administration of trinitrobenzenesulfonic acid [TNBS]. The animals were sacrificed on day 5 after TNBS administration and colonic and ileal samples were used for estimation of myeloperoxidase [MPO] activity, malondialdehyde [MDA] concentration, histologic examination and transmission electron microscopy. TNBS caused a significant reduction in body weight and an increase in MPO activity in colonic, but not in the ileal samples in animals with colitis. MDA levels were increased both in inflamed colon and the uninflamed ileal segments in colitis. Electron microscopy revealed swelling of mitochondria with broken cristae and disruption of the inner membrane. Colitis also caused fragmentation of the ER with loss of ribosomes and swelling of the Golgi apparatus with distended vesicles in both smooth muscle and epithelial cells in the ileal and colonic segments. These changes were absent in the control rats without colitis. These findings demonstrate ultrastructural deformities in both the mucosa and smooth muscle in inflamed and uninflamed regions of the gastrointestinal tract in experimental colitis. The structural changes in mitochondria are responsible for reduced ATP, while abnormalities in the ER and the Golgi apparatus may explain a generalized effect on protein synthesis, trafficking and targeting mechanisms, and may account for physiological changes seen in experimental colitis

light and scanning electron microscope study of the albino rat ileum after its partial obstruction

Induction of some sort of obstruction could be resorted to as a definitive line of management of some cases of short- bowel syndrome [SBS]. The goal of this study was to clarify the histological and morphometric alterations in the albino rat ileum after its surgically induced partial obstruction. Thirty adult male albino rats [240-250 gm] were used in this investigation. They were divided into two equal groups: control and experimental. Small pieces of the ileum of the control and experimental animals were processed for histological and scanning electron microscope study. The ileum of the experimental animals proximal to the site of obstruction showed an apparent enlargement in the Peyer's patches and an increase in the thickness of both the mucosa and muscle layers. The villi showed significant elongation and thickening. Both widening and deepening of the crypts were detected. There were an apparent increment in the goblet cell number and in the lymphocytic infiltration in both the corium and submucosa. Scanning electron microscopic examination, the microvilli showed scattered areas of shortening and irregular orientation. The surface was more frequently broken by goblet cell orifices. It could be concluded that partial ileal obstruction resulted in hypertrophy of the ileal wall with considerable structural alterations oral to the obstruction site. Thus, a procedure apparently increased the absorptive surface area together with slowing the intestinal transit. These effects could support putting forward this technique into consideration as one of the suggested lines of treatment of some cases of short bowel syndrome to eliminate the patients need for the parenteral nutrition and all its associated complications

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.

Disruption of the intestinal barrier and bacterial translocation in an experimental model of intestinal obstruction

Clinical evidence and the use of experimental models in laboratory animals indicate that the intestine is a reservoir of microorganisms that can cause systemic infection in the human. The purpose of this work was to study the possible effect of intestinal obstruction (IO) on the mechanical and chemical barriers that bring protection against microorganisms crossing from the intestinal lumen towards the systemic tissues. We demonstrated that 24 hours after IO, histological and ultrastructural alterations do occur, seriously compromising the structure of the intestinal barrier in 100 per cent of the studies animals. Likewise, it was observed that during the same period, microorganisms translocation from intestine to the peritoneal cavity and liver (100 and 80 per cent respectively) occurred. The lungs were spared. Changes observed in the intestinal epithelium are related to a process similar to that produced by intestinal ischemia: mitochondrial destruction, with subsequent decrease of its capacity to supply eb

Colectomia total e ileopapilorretoanastomose término-terminal ou por telescopagem / Total colectomy and ileorrectal anastomosis terminal

No estudo experimental aqui apresentado, submetemos 20 cäes à ligadura do ramo terminal da artéria ileal. Os animais foram divididos em dois grupos: A (10 cäes) e B (10 cäes), que foram observados e sacrificados, após 48 e 96 horas, respectivamente. Todo o segmento ileocecal foi retirado para estudo macro e microscópico do íleo terminal e da papila ileocecal. Pudemos verificar que a ligadura arterial näo comprometeu a viabilidade do íleo terminal e da papila ileocecal. O aspecto microscópico do íleo terminal após 96 horas, mostrou-se normal em 70 por cento. O aspecto microscópico da papila ileocecal após 96 horas, mostrou-se normal em 90 por cento. No plano clínico estudamos 12 pacientes submetidos à colectomia total e anastomose ileorretal preservando-se a papila ileocecal (ileopapilorretoanastomose). Os pacientes foram divididos em dois grupos: A (seis pacientes) foi realizada uma telescopagem ileorretal e no grupo B (seis pacientes) uma anastomose ileopapilorretal término-terminal. No grupo A optou-se pela mucosectomia parcial em dois pacientes e total em um; no grupo B optou-se pela mucosectomia parcial em um paciente. Os pacientes foram observados após sete, 15, 40, 60 e 365 dias quando procuramos avaliar: número de evacuaçöes diárias; aspecto macroscópico do períneo; aspectos macro e microscópico das mucosas do reto, íleo terminal e da papila ileocecal e aspectos radiológicos da anastomose ileorretal...

Histochemical, ultrastructural and stereological study of the mucosa of ileal neobladder

This study was undertaken to determine the effect of chronic urinary diversion on the morphology of intestine. Biopsies from control ileum and ileal neobladder taken after 6 and 24 months postoperatively were examined by light and transmission electron microscopy with employing the recent stereological methods. The histochemical changes observed were diminution of alkaline phosphatase and succinic dehydrogenase activity in the mucosa of the ileal neobladder compared with control ileum. Moreover, the light microscopical examination of the ileal neobladder showed mucosal atrophy evidenced by shortening of villi and increasing the depth of crypts. Quantitative findings confirmed the previous qualitative atrophic changes where the villous to crypt ratio changed from 3.9 in control ileum to 2.2 and 1.2 in the ileal neobladder after 6 and 24 months, respectively. The ultrastructural changes in the ileal neobladder consisted of a reduction in the number and height of the microvilli, increased lysosomal features and the activity of muciparous cells and a loss of the normal polarized deposition of the cytoplasmic organelles. Both quantitative and qualitative findings confirmed that the older the neobladder, the more severe the changes found

Estudo morfológico da mucosa de um segmento de colo distal interposto entre cotos de intestino delgado após ressecçäo de oitenta por cento de seu comprimento: estudo experimental, com microscopia eletrônica de varredura, em ratas / Morphological study of the mucosa of a segment of the distal colon interposed between stumps of the small intestine after resection of 80 percent of its length: experimental study, with scanning electron microscopy, in female rats

O objetivo deste trabalho foi verificar em estudo com microscopia eletrônica de varredura, quais as eventuais alteraçöes morfológicas ocorridas na mucosa do intestino, seguindo o modelo experimental sobre a adaptaçäo do colo distal interposto entre cotos de intestino delgado após ressecçäo de oitenta por cento de seu comprimento, em ratos com noventa dias de observaçäo. Foram selecionados segmentos de jejuno, íleo, colo normal de cinco animais de controle e segmentos de jejuno a montante e íleo a jusante do colo interposto e do próprio colo interposto em cinco animais operados. Os resultados mostraram que ocorre um aumento de celularidade em todos os segmentos do grupo operado, com aumento das dimensöes das vilosidades no delgado. No segmento de colo interposto o aumento de celularidade leva alteraçöes do padräo morfológico em relaçäo ao normal, com modificaçöes dos óstios das criptas intestinais, modificaçöes nas características da camada de muco do colo e principalmente surgem formaçöes verrucosas na camada epitelial como projeçöes celulares ou brotos celulares. Embora a camada epitelial do colo interposto sofra modificaçöes, elas porém näo propiciam dados suficientes para denominá-las como "delgadizaçäo"

Estudo de ilcostomia em cäes: aspecto funcional e microscopia óptica e eletrônica de varredura / Ileostomy in dogs: functional aspects and optic and scanning electronic microscopy

A ileostomia continente tem sido aspiraçäo permanente dos pesquisadores. Diversas técnicas têm sido propostas. Bem poucas preencheram alguns requisitos, porém nenhum atingiu o objetivo desejado e o problema continua por solucionar. Os autores com base em estudo experimental em dois grupos de cäes submetidos a ileostomia, com e sem a preservaçäo do piloro ileal, observaram no pós-operatório os seguintes parâmetros: peso dos cäes, avaliaçäo clínica dos animais (diarréia, infecçäo da ferida cirúrgica e desnutriçäo), volume das excreçöes ileais, eletrólitos (Na+, Cl- e K+), o pH das excreçöes ileais e óbitos. Nos pós-sacrifício dos animais analisaram ao microscópio eletrônico de varredura a presença de muco, nitidez da superfície da mucosa, aspecto geral da mucosa do íleo terminal; ao microscópio óptico a infiltraçäo neutrofílica, aumento do número de células caliciformes, hipotrofia e determinaçäo da altura das vilosidades ileais. Essa técnica de feitura de ileostomia com a preservaçäo do piloro ileal, apresenta grandes vantagens sobre outros procedimentos, principalmente no que tange ao piloro ileal, que atenua nitidamente a perda das excreçöes, como ficou demostrado pela análise estatística significativa dos resultados das variáveis do experimento. A conservaçäo do piloro ileal, portanto, tem uma açäo efetiva no controle do esvaziamento ileal, protegendo até sua mucosa de alteraçöes histopatológicas graves