Characterization of immune and enteric systems of broilers after imunosupression with dexamethasone

Background: Bursa of Fabricius (BF) and the thymus are primary lymphoid organs of poultry and play a major role in avian immunity. Enteric system is also involved in immunity. Several pathologic conditions directly impact BF and thymus size, and also affect intestinal parameters. Besides, there are several immune system depressor agents which affect birds. The selection of glucocorticoid as inducer of immunosuppression is applied in many experiments; however there are few studies that are applied to the reality in the field. In this context, the aim of this study was to evaluate the effects of dexamethasone as an inducer of immunosuppression on lymphoid organs and microscopic structures of the jejunum.Materials, Methods & Results: One-day-old chicks were used as a control group (n = 8) and the treated group (n = 25) received intramuscular dexamethasone on 21, 23, 24 and 26 day-old. Control birds and treated birds were euthanized 8, 16, 24, 32 and 40 h after inoculation; four control birds and six treated birds were euthanized on the eighth day after the last inoculation. Thymus, BF and jejunum were collected during the necropsy. The selected organs were processed, stained with hematoxylin and eosin and photographed. The BF and thymus cuts were evaluated by three histopathologists to determine the depletion score. Ten villi of each jejunum were evaluated for width and length of villi, depth crypt, microvillus length, enterocyte length of each villus, and wall thickness. Treated birds presented a mean weight lower than control group during all the experiment. The mean weight and the relative weight of the BF and thymus of control birds were significantly higher than treated ones. The lymphocyte depletion in BF and thymus scores differed significantly between groups, being higher in the group challenged with dexamethasone. There were no significant differences between groups for depth of crypt, height of core and height of microvilli.[...]

Characterization of immune and enteric systems of broilers after imunosupression with dexamethasone

Background: Bursa of Fabricius (BF) and the thymus are primary lymphoid organs of poultry and play a major role in avian immunity. Enteric system is also involved in immunity. Several pathologic conditions directly impact BF and thymus size, and also affect intestinal parameters. Besides, there are several immune system depressor agents which affect birds. The selection of glucocorticoid as inducer of immunosuppression is applied in many experiments; however there are few studies that are applied to the reality in the field. In this context, the aim of this study was to evaluate the effects of dexamethasone as an inducer of immunosuppression on lymphoid organs and microscopic structures of the jejunum.Materials, Methods & Results: One-day-old chicks were used as a control group (n = 8) and the treated group (n = 25) received intramuscular dexamethasone on 21, 23, 24 and 26 day-old. Control birds and treated birds were euthanized 8, 16, 24, 32 and 40 h after inoculation; four control birds and six treated birds were euthanized on the eighth day after the last inoculation. Thymus, BF and jejunum were collected during the necropsy. The selected organs were processed, stained with hematoxylin and eosin and photographed. The BF and thymus cuts were evaluated by three histopathologists to determine the depletion score. Ten villi of each jejunum were evaluated for width and length of villi, depth crypt, microvillus length, enterocyte length of each villus, and wall thickness. Treated birds presented a mean weight lower than control group during all the experiment. The mean weight and the relative weight of the BF and thymus of control birds were significantly higher than treated ones. The lymphocyte depletion in BF and thymus scores differed significantly between groups, being higher in the group challenged with dexamethasone. There were no significant differences between groups for depth of crypt, height of core and height of microvilli.[...](AU)

Lactobacillus plantarum culture supernatants improve intestinal tissue exposed to deoxynivalenol.

In the present study, histological, morphometrical and ultrastructural analysis were performed to investigate intestinal mucosa changes in piglets exposed to deoxynivalenol alone or associated with two strains of Lactobacillus plantarum and the respective culture supernatants. Jejunal explants were incubated for 4h in culture medium with a) only culture medium (DMEM, control group), b) deoxynivalenol (DON, 10µM), c) heat-inactivated Lactobacillus plantarum strain1 - LP1 (1.1×108 CFU/ml) plus DON, d) heat-inactivated Lactobacillus plantarum strain2-LP2 (2.0×109 CFU/ml) plus DON, e) heat-inactivated Lactobacillus plantarum strain1 culture supernatant (CS1) plus DON, and f) heat-inactivated Lactobacillus plantarum strain1 culture supernatant (CS1) plus DON. Explants exposed to DON and DON plus LP1 and LP2 showed a significant increase in histological changes (mainly villi atrophy and apical necrosis) and a significant decrease in villi height when compared to unexposed explants. However, explants treated with CS1+DON and CS2+DON remained similar to the control group both in histological and morphometrical aspects. DON also induced a significant decrease in goblet cell density compared to control whereas CS1+DON treatment induced an increase in the number of goblet cells in comparison to DON explants. In addition, ultrastructural assessment showed control, CS1+DON and CS2+DON explants with well delineated finger shape villi, meanwhile DON-treated, LP1+DON and LP2+DON explants showed a severe villi atrophy with leukocytes exudation on the intestinal surface. Taken together, our results indicate that the culture supernatant treatment reduced the toxic effects induced by DON on intestinal tissue and may contribute as an alternative strategy to reduce mycotoxin toxicity.

Dynamic regulation of extracellular ATP in Escherichia coli.

We studied the kinetics of extracellular ATP (ATPe) in Escherichia coli and their outer membrane vesicles (OMVs) stimulated with amphipatic peptides melittin (MEL) and mastoparan 7 (MST7). Real-time luminometry was used to measure ATPe kinetics, ATP release, and ATPase activity. The latter was also determined by following [32P]Pi released from [γ-32P]ATP. E. coli was studied alone, co-incubated with Caco-2 cells, or in rat jejunum segments. In E. coli, the addition of [γ-32P]ATP led to the uptake and subsequent hydrolysis of ATPe. Exposure to peptides caused an acute 3-fold (MST7) and 7-fold (MEL) increase in [ATPe]. In OMVs, ATPase activity increased linearly with [ATPe] (0.1-1 µM). Exposure to MST7 and MEL enhanced ATP release by 3-7 fold, with similar kinetics to that of bacteria. In Caco-2 cells, the addition of ATP to the apical domain led to a steep [ATPe] increase to a maximum, with subsequent ATPase activity. The addition of bacterial suspensions led to a 6-7 fold increase in [ATPe], followed by an acute decrease. In perfused jejunum segments, exposure to E. coli increased luminal ATP 2 fold. ATPe regulation of E. coli depends on the balance between ATPase activity and ATP release. This balance can be altered by OMVs, which display their own capacity to regulate ATPe. E. coli can activate ATP release from Caco-2 cells and intestinal segments, a response which in vivo might lead to intestinal release of ATP from the gut lumen.

Comparative Analysis of the Morphology, Ultrastructure, and Glycosylation Pattern of the Jejunum and Ileum of the Wild Rodent Lagostomus maximus.

Morphological and histochemical analyses were performed to characterize the histology, ultrastructure, and glycosylation pattern of the jejunum and ileum of the wild rodent Lagostomus maximus. Enterocytes, goblet cells, Paneth cells, and enteroendocrine cells were identified in both intestinal epithelia. Two morphological types of enterocytes were identified only in the ileum based on their cytoplasm electron density. Although the histological and ultrastructural examination showed that the epithelia of both anatomical regions were morphologically similar, a certain specialization in their secretory products was evident. The glycosylation pattern of the jejunum and ileum was characterized in situ by histochemical and lectin histochemical methods. Histochemical results revealed the presence of carboxylated and sulfated gycoconjugates in both regions, although sulfomucins were clearly prevalent in the ileum. Sialic acid was highly O-acetylated and particularly abundant in the jejunum. The KOH/PA*/Bh/PAS technique evidenced a more intense histochemical reaction in the jejunal than in the ileum goblet cells, demonstrating a reduction of neutral mucin secretion in the distal small intestine. Further specific differences were revealed by lectin histochemistry. These data evidenced that the nature of mucus varies at different anatomical regions, probably adapted to physiological requirements.

Deoxynivanelol and fumonisin, alone or in combination, induce changes on intestinal junction complexes and in E-cadherin expression.

Fusariotoxins such as fumonisin B1 (FB1) and deoxynivalenol (DON) cause deleterious effects on the intestine of pigs. The aim of this study was to evaluate the effect of these mycotoxins, alone and in combination, on jejunal explants from piglets, using histological, immunohistochemical and ultrastructural assays. Five 24-day old pigs were used for sampling the explants. Forty-eight explants were sampled from each animal. Explants were incubated for 4 hours in culture medium and medium containing FB1 (100 µM), DON (10 µM) and both mycotoxins (100 µM FB1 plus 10 µM DON). Exposure to all treatments induced a significant decrease in the normal intestinal morphology and in the number of goblet cells, which were more severe in explants exposed to DON and both mycotoxins. A significant reduction in villus height occurred in groups treated with DON and with co-contamination. Expression of E-cadherin was significantly reduced in explants exposed to FB1 (40%), DON (93%) and FB1 plus DON (100%). The ultrastructural assay showed increased intercellular spaces and no junction complexes on enterocytes exposed to mycotoxins. The present data indicate that FB1 and DON induce changes in cell junction complexes that could contribute to increase paracellular permeability. The ex vivo model was adequate for assessing intestinal toxicity induced by exposure of isolated or associated concentrations of 100 µM of FB1 and 10 µM of DON.

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...

Ultrastructural development of the small intestinal mucosa in the embryo and turkey poult: A light and electron microscopy study.

The potential for growth and feed efficiency in turkey poults directly correlates with the early development of the intestinal epithelium. Although the metabolic aspects of enteric maturation have been studied, little is known about the ultrastructural development of the enteric epithelium in the turkey embryo and poult. Hence, the objective of this study was to document the morphological and ultrastructural development of the jejunum mucosa in turkeys, from 15 d of incubation (embryonic day; E) to 12 d posthatch. Intestinal samples from 4 embryos or poults were collected and analyzed by light and electron microscopy (transmission and scanning). In addition, amniotic fluid volume was determined in 6 eggs from E15 to E25. Longitudinal previllus ridges at E15 gradually formed zigzag patterns that led to the formation of 2 parallel lines of mature villi by E25. The volume of amniotic fluid was rapidly depleted as the embryo swallowed it between E19 and E25. During this period, a major increase occurs in villus height, the apical end of epithelial cells is gradually tightened by the junctional complex, and mature goblet cells are visible at the apical end of villi. Villus height steadily increases until reaching a plateau at 8 d. Villi morphology shifts gradually from finger-like projections before hatch to leaf-like projections by 12 d. At this age, the enteric epithelium is in intimate association with microbes such as segmented filamentous bacteria. The profound morphological adaptations of the turkey gut epithelium in response to amniotic fluid swallowing before hatch, and dietary factors and bacteria after hatch, demonstrate the plasticity of the enteric epithelium at this time. Hence, the supplementation of enteric modulators before hatch (in ovo feeding) and after hatch has the potential to shape gut maturation and enhance the growth performance of turkey poults.

Morphoquantitative study of the submucous plexus (of Meissner) of the jejunum-ileum of young and old guinea pigs.

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 cm(2) 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/cm(2) average in young animals and 2918±120.70 neurons/cm(2) 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% of the neurons with aging.

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.

B cells are involved in the modulation of pathogenic gut immune response in food-allergic enteropathy.

Food enteropathies involve uncontrolled or hypersensitivity reactions to ingested nutrients and may result in IgE and T-helper type 2 (Th2) responses as in food allergy. However, the precise role of B cells in the development of food enteropathies remains uncertain. In this work, we used B cell-deficient mice (B KO) and a model of peanut sensitization to examine the involvement of B lymphocytes in the pathogenesis of food allergy. Results showed that priming of wild-type (WT) mice with peanut proteins induced specific IgG1 and IgE responses in serum, with edema, tissue destruction, epithelial exulceration and inflammatory infiltrate in the gut of sensitized and challenged (S + Peanut) WT animals. In contrast, there was no sera immunoglobulin detection and absence of tissue destruction in the gut of B KO mice, which presented moderate inflammatory infiltrate and villous enlargement after peanut challenge. These animals presented marked decrease in IL-4 and TNF-alpha and high levels of IL-10, TGF-beta, IL-12p40 and IFN-gamma mRNA in the gut. Moreover, the expression of CCL5, CCL11 and CXCL1 was reduced in the gut of B KO mice, in contrast to elevated messages of CCL2 or similar detection of Th1-related chemokines in S + Peanut WT mice. Finally, we provided evidence that B cells are necessary to the development of food-related enteropathies and induction of gut inflammation during allergic reactions to food.

Alterations of neurokinin receptors and interstitial cells of Cajal during and after jejunal inflammation induced by Nippostrongylus brasiliensis in the rat.

Substance P (SP) and its receptors NK1 and NK2 are widely expressed in the intestinal wall by neurones, interstitial cells of Cajal (ICC) and smooth muscle cells. Changes in SP and/or its NK receptors have been documented during experimental inflammation in animals or inflammatory bowel diseases in humans, but the data concern the acute phase of the inflammatory process. We determined immunohistochemically whether NK receptors and SP were altered in the muscle coat during jejunal inflammation induced by the nematode Nippostrongylus brasiliensis and whether these alterations persisted when inflammation had spontaneously resolved 30 days postinfection. An ultrastructural analysis was also conducted on ICC, nerves and muscle. At day 14, when inflammation peaked, there was a reduction in NK1 receptors in myenteric neurones and in SP-immunoreactive nerve endings. There were also ultrastructural anomalies in synaptic vesicles and NK2 receptor loss in the circular muscle layer. The SP decrease persisted at day 30, whereas neurones and circular muscle cells re-expressed NK1 and NK2 receptors, respectively. The ICC at the deep muscular plexus, located near to the inflammatory site, underwent alterations leading to their complete loss at day 30. These morphological changes are probably associated with impairment in tachykinergic control of jejunal functions leading to the alterations of motility and sensitivity to distension already described in these animals.

Chemokine and cytokine expression in murine intestinal epithelium following Nippostrongylus brasiliensis infection.

Infection of mice with the nematode parasite Nippostrongylus brasiliensis results in a well characterized intestinal mastocytosis with intraepithelial migration of mucosal mast cells (MMC). The molecules mediating this response are unknown. We examined expression of several putative mast cell chemoattractants in intestinal epithelium following N. brasiliensis infection. Expression of the chemokines monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-1 alpha (MIP-1alpha), RANTES (regulated on activation normal T-cell expressed and secreted), fractalkine, and thymocyte expressed chemokine (TECK); and the cytokines stem cell factor (SCF) and transforming growth factor beta1 (TGFbeta1), was constitutive and no alteration was detected following infection. MCP-1 expression was also constitutive but at much lower levels and increased expression was detected on days 7 and 14 postinfection. Expression of MCP-1 in whole jejunum was at much higher levels than in epithelium. Constitutive expression of MCP-1, MIP-1alpha and TGFbeta1 was also detected in cultured bone marrow-derived homologues of MMC. In an intestinal epithelial cell line (CMT-93), there was constitutive expression of SCF, TGFalpha1, fractalkine and MCP-1. The results show that, in vivo, epithelium is a potentially important source of mast cell chemoattractants.

Scanning electron microscopy of the jejunum in children with protein-energy malnutrition.

Samples of jejunum from seven children dying from severe oedematous malnutrition were fixed with intralumenal glutaraldehyde in combination with external immersion, using a rapid autopsy protocol. Selected areas were post-fixed in osmium tetroxide, dehydrated with an ethanol series and critical point dried. After mounting on aluminium stubs, specimens were sputter-coated with a thin layer of gold and palladium. Scanning electron microscopy showed patterns ranging from subtotal villous atrophy, through partial villous atrophy with low ridges, to longer, taller ridges with cerebriform convolutions. In one case, villous height had been interpreted as normal on light microscopy. The scanning electron microscope revealed that the mucosa in this case was composed of ridges and leaf forms. Processing for scanning electron microscopy is simple. The specimens can be viewed and photographed in a relatively short time. In addition, the three-dimensional record provides clarification of ambiguities which often arise from routine histological sections.

Ultrastructural study of enteropathogenic Escherichia coli Olllab: H2 infection in an infant with acute diarrhea

Escherichia coli enteropatogênica clássica é o mais importante agente etiológico como causa de diarréia aguda nos países subdesenvolvidos, especialmente nos lactentes menores de uma ano de idade. As cepas de Escherichia coli enteropatogênica sao capazes de provocar profundas alteraçoes no citoesqueleto do enterócito, as quais sao denominadas aderência e apagamento das microvilosidades do enterócito, as quais sao denominadas aderência e apagamento das microvilosidades, associadas à formaçao de pedestais. Descreve-se uma cepa de Escherichia coli enteropatogênica 0111ab:H2 isolada das fezes de um lactente com diarréia aguda, no 11§ dia de doença, a qual provocou lesoes de aderência e apagamento das microvilosidades do enterócito e inclusive penetrou no interior do citoplasma do mesmo, assim como invadiu células HeLa em cultura de tecido in vitro e da mesma forma no teste da alça ileal ligada de coelho in vivo, todas as lesoes detectadas em estudo ultra-estrutural. Esta observaçao demonstra que lesoes graves da mucosa do intestino delgado causadas por uma cepa de Escherichia coli enteropatogênica 0111ab:H2 podem ocorrer ainda em um estágio precoce da infecçao entérica.

Casein, hydrolyzed casein, and amino acids that simulate casein produce the same extent of mucosal adaptation to massive bowel resection in adult rats.

We determined the effect of the extent of protein polymerization on the intestinal hyperplastic adaptation of adult male Wistar rats after 80% resection of the jejunal-ileal segment. Rats received one of four chemically defined solid diets prepared by using casein, two casein hydrolysates of different peptide size distributions, or free amino acids simulating casein and identical in all other components for 12 d, starting 3 d after surgery. Semipaired feeding was used to ensure that the same quantity of food was ingested by each group and as a consequence, nitrogen and energy intakes were reduced to 63% of that obtained with ad libitum feeding of the casein diet to intact rats. No significant differences were demonstrable in food ingestion, weight gain, nitrogen balance, or morphometric data for the remaining jejunal and ileal segments (number of cells/villus, number of cells/crypt, and crypt cell mitosis rate). These data demonstrate that the extent of polymerization of the protein nitrogen source did not affect the hyperplastic adaptative process of the rat. Additional studies in humans are necessary to determine whether intact protein diets can be used first as a nitrogen source in nutritional support of patients with a nonspecific hyperplastic response to surgical resection before the use of expensive hydrolysates and the more expensive amino acid mixtures.

Ultrastructural study of enteropathogenic Escherichia coli O111ab:H2 infection in an infant with acute diarrhea.

Enteropathogenic Escherichia coli is the most important cause of acute diarrhea in developing countries, specially in infants under one year of age. Enteropathogenic Escherichia coli strains are able to induce profound cytoskeletal alterations in the enterocyte known as attaching and effacing lesions, associated with the formation of cuplike pedestals. We report an Enteropathogenic Escherichia coli O11ab:H2 strain isolated from an infant with acute diarrhea, on the eleventh day of disease, that caused attaching and effacing lesion and penetrated the enterocyte, as well as invaded the HeLa cell tissue culture in vitro and the rabbit ileal loop assay in vivo, in the ultrastructural study. This observation indicates that the severe lesions of the small bowel caused by an enteropathogenic Escherichia coli O111ab:H2 strain can occur even in the early stages of the infection.

Jejunal absorption of trypsin in rat and guinea pig.

Gut absorption is one of the first requirements for the study of the mechanism of a possible anti-inflammatory action of proteases, such as orally administered trypsin. Porcine trypsin absorption was studied in isolated jejunal loops of rats (female Holtzman and male Wistar) and guinea pigs (males) by open-loop perfusion. Trypsin was dissolved in Tyrode solution and the solution perfused at a rate of 0.5 ml/min, at 37 degrees C. Trypsin activity, total protein, and sodium and potassium concentrations were assayed in the jejunal effluent; the values were unchanged throughout the experiments, which lasted 45 to 120 min. Using a high sensitivity ELISA (i.e. pg/ml), trypsin absorption could be demonstrated by determination of the enzyme in the mesenteric venous blood (samples of 0.5 ml); the enzyme concentration increased with time of perfusion. The linear range-specificity for intact trypsin varied from 1 to 500 ng/well. In this assay polyclonal antibodies prepared against trypsin-TLCK were utilized. Whereas trypsin concentration in the perfused lumen was practically constant at 0.12 mg/ml, the concentration of absorbed trypsin in mesenteric vein blood increased from about 100 ng/ml at time zero to 1.8 micrograms/ml, after 45 min of perfusion. Histological and ultrastructural examination of the jejunal mucosa before and after perfusion revealed that the brush-border, basal membrane, and junctional complexes were fully preserved, thus eliminating the possibility that trypsin might have destroyed the structures, thereby reaching the blood circulation. The present data indicate that micrograms quantities of trypsin were absorbed by the isolated jejunal loop of the rat.

Jejunal absorption of trypsin in rat and guinea pig

Gut absorption is one of the first requirements for the study of the mechanism of a possible anti-inflammatory action of proteases, such as orally administered trypsin. Porcine trypsin absorption was studied in isolated jejunal loops of rats (female Holtzman and male Wistar) and guinea pig (males) by open-loop perfusion. Trypsin was dissolved in Tyrode solution and the solution perfused at a rate of 0.5 ml/min, at 37§C. Trypsin activity, total protein, and sodium and potassium concentrations were assayed in the jejunal effluent; the values were unchanged throughout the experiments, which lasted 45 to 120 min. Using a high sensitivity ELISA (i.e. pg/ml), trypsin absorption could be demonstrated by determination of the enzyme in the mesenteric venous blood (samples of 0.5 ml); the enzyme concentration increased with time of perfusion. The linear range-specificity for intact trypsin varied from 1 to 500 ng/well. In this assay polyclonal antibodies prepared against trypsin-TLCK were utilized. Whereas trypsin concentration in the perfused lumen was practically constant at 0.12 mg/ml, the concentration of absorbed trypsin in mesenteric vein blood increased from about 100 ng/ml at time zero to 1.8 µg/ml, after 45 min of perfusion. Histological and ultrastructural examination of the jejunal mucosa before and after perfusion revealed that the brush-border, basal membrane, and junctional complexes were fully preserved, thus eliminating the possibility that trypsin might have destroyed the structures, thereby reaching the blood circulation. The present data indicate that µg quantities of trypsin were absorbed by the isolated jejunal loop of the rat

Familial enteropathy with villous edema and immunoglobulin G2 subclass deficiency.

We describe a familial form of recurrent acute, life-threatening secretory diarrhea associated with distinctive jejunal histologic changes and IgG2 subclass deficiency. Symptoms begin abruptly with anorexia and vomiting, and progress within hours to massive secretory diarrhea and shock with profound neutropenia and hypoproteinemia, including hypoalbuminemia and hypogammaglobulinemia. Affected survivors recover quickly and thereafter grow and develop normally. Biopsy specimens obtained during remission from 3 adults and 11 children show club-shaped jejunal villi broadened by edema and histiocytes with imbibed fluid; the overlying intestinal epithelium and brush border appear normal, but the basement membrane is interrupted in some areas. No characteristic microorganisms have been identified in association with the syndrome. Clinical manifestations cease in the second decade, but the abnormal jejunal histologic pattern persists into adult life. Female and male patients are equally affected, although all fatal cases have been in female subjects. Inheritance appears dominant with variable penetrance: one family member without a history of diarrhea has characteristic biopsy findings and another appears to be an obligate carrier with normal biopsy findings. Affected individuals have a reduced serum concentration of IgG2. We believe that this familial enteropathy is a unique entity, not previously described.