Diphenyleneiodonium Inhibits Apoptotic Cell Death of Gastric Epithelial Cells Infected with Helicobacter pylori in a Korean Isolate

NADPH oxidase produces a large amount of reactive oxygen species (ROS) in Helicobacter pylori (H. pylori)-induced gastric epithelial cells. Even though ROS mediate apoptotic cell death, direct involvement of NADPH oxidase on H. pylori-induced apoptosis remains unclear. Besides, H. pylori isolates show a high degree of genetic variability. The predominant genotype of H. pylori in Korea has been reported as cagA+, vacA s1b, m2, iceA genotype. Present study aims to investigate whether NADPH oxidase-generated ROS mediate apoptosis in human gastric epithelial AGS cells infected with H. pylori in a Korean isolate. AGS cells were pretreated with or without an NADPH oxidase inhibitor diphenyleneiodonium (DPI) and cultured in the presence of H. pylori at a bacterium/cell ratio of 300:1. Cell viability, hydrogen peroxide level, DNA fragmentation, and protein levels of p53, Bcl-2, and Bax were determined. Results showed that H. pylori inhibited cell viability with the density of H. pylori added to the cells. Inhibition of NADPH oxidase by DPI suppressed H. pylori-induced cell death, increased hydrogen peroxide, DNA fragmentation, and the ratio of Bax/Bcl-2, and p53 induction in AGS cells dose-dependently. The results suggest that targeting NADPH oxidase may prevent the development of gastric inflammation associated with H. pylori infection by suppressing abnormal apoptotic cell death of gastric epithelial cells.

The brainstem localization of gastric preganglionic parasympathetic neurons in the Agouti (Dasyprocta leporina) / Localización del tronco encefálico de las neuronas parasimpáticas preganglionares gástricas en el agutí (Dasyprocta leporina)

This study was designed to determine qualitatively, the source of gastric vagal nerve fibres in the Agouti. A total of 18 male and female adult agoutis were used for the present investigation. Following anaesthesia, laparotomy was performed and the stomach exteriorized. Multiple intramuscular injections of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) were then made into different areas of the stomach in the experimental animals. The control animals were divided into four groups of two animals each. The first group had intraperitoneal injection of the tracer, the second had intramuscular injection of normal saline, the third group had injection of tracer into the hepatic portal vein and the last group had injection of the tracer into the gastric walls followed immediately by bilateral vagotomy. Following a survival period offive to seven days, the animals were sacrificed by transcardial perfusion, first with normal saline followed by fixative and finally with 20% buffered sucrose. Following perfusion, the brainstem was extracted from the brain, immersed in 20% buffered sucrose and kept refrigerated overnight for cryoprotection. The brainstems were subsequently sectioned serially, processed for WGA-HRP neurohistochemistry and then analysed under light and dark-field illuminations. The analysis of the sections taken from the experimental animals revealed bilateral presence of WGA-HRP labelled neurons in the dorsal motor nucleus of the vagus nerve (DMNV) and the nucleus ambiguus (nA) of the medulla oblongata. No labelled neurons were seen in any of the sections taken from the control animals. The implications of the findings are discussed.

Este estudio fue diseñado para determinar cualitativamente el origen de las fibras gástricas del nervio vago en el agutí. Un total de 18 agutíes adultos masculinos y femeninos fueron utilizados para la presente investigación. Después de la anestesia, se realizó una laparotomía y se sacó el estómago al exterior. Luego se hicieron múltiples inyecciones intramusculares de aglutinina de germen de trigo con peroxidasa de rábano (WGA-HRP) en diferentes áreas del estómago de los animales experimentales. Los animales del control fueron divididos en cuatro grupos de dos animales cada uno. Al primer grupo se le puso una inyección intraperitoneal del marcador; al segundo se le administró una inyección intramuscular de solución salina normal; al tercer grupo se le inyectó el marcador en la vena porta hepática; y al último grupo se le puso la inyección del marcador en las paredes gástricas, seguida inmediatamente por una vagotomía bilateral. Tras un periodo de supervivencia de cinco a siete días, los animales fueron sacrificados por perfusión transcardíaca, primero con solución salina normal, seguida de fijador, y finalmente con sacarosa tamponada al 20%. Después de la perfusión, el tronco encefálico fue extraído del cerebro, inmerso en sacarosa tamponada al 20%, y mantenido en refrigeración durante la noche para su crioprotección. Los tronos encefálicos fueron luego seccionados en serie, procesados para para el análisis neuro-histoquímico mediante aglutinina de germen de trigo con peroxidasa de rábano, y analizados entonces bajo iluminaciones de campo de luz y campo oscuro. El análisis de las secciones tomadas de animales experimentales reveló la presencia bilateral de neuronas etiquetadas WGA-HRP en el núcleo motor dorsal del nervio vago (DMNV) y en el núcleo ambiguo (nA) de la médula oblonga. No se observaron neuronas etiquetadas en ninguna de las secciones tomadas de los animales de control. Se discuten las implicaciones de los hallazgos.

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...

In silico prediction of exposure amino acid sequences of outer inflammatory protein A of Helicobacter pylori for surface display on Eschierchia coli.

BACKGROUND: Outer inflammatory protein A (OipA) is an outer membrane protein of Helicobacter pylori that is involved in inducing IL-8 and intracellular signaling. In this study, we have predicted exposure amino acid sequences of OipA for insertion in permissive sites of CstH subunit of Eschierchia coli CS3 pilli for bacterial surface display. MATERIALS AND METHODS: Databases: National Center for Biotechnology Institute and Protein Data Bank. Servers: PHD, SABLE, GOR 4, SignalP3.0, TBBpred, PRODIV-TMHMM, TMRPres2D, CPH Models, PHYRE, GETAREA, VADAR, Pep state and pep window. Software: Swiss PDB viewer and Discovery studio. RESULTS: In silico prediction of exposure amino acid sequences of OipA led to detection of six sequences of amino acid, 76-87, 106-112, 170-182, 222-230, 242-258, and 278-290. These sequences inserted between amino acid sequences 66-67, 100-101, and 109-110 of CstH that were predicted by Eskandari et al. as permissive sites of CstH. CONCLUSION: OipA has the ability to induce IL-8 from gastric epithelial cells and some papers are mentioned that this outer membrane protein involve to attachment and intracellular signaling. Receptor of OipA and adhesion motifs on this protein is unknown. Detection of exposure motifs aids to recognition of adhesion motifs and receptor of OipA on gastric epithelial cells. In this study, we have predicted exposure amino acid sequences for insert to subunit CstH of CS3 pilli E. coli for surface display.

DNA Methylation and Expression Patterns of Key Tissue-specific Genes in Adult Stem Cells and Stomach Tissues

CpG-island margins and non-island-CpG sites round the transcription start sites of CpG-island-positive and -negative genes are methylated to various degrees in a tissue-specific manner. These methylation-variable CpG sites were analyzed to delineate a relationship between the methylation and transcription of the tissue-specific genes. The level of tissue-specific transcription was estimated by counting the number of the total transcripts in the SAGE (serial analysis of gene expression) database. The methylation status of 12 CpG-island margins and 21 non-island CpG sites near the key tissue-specific genes was examined in pluripotent stromal cells obtained from fat and bone marrow samples as well as in lineage-committed cells from marrow bulk, stomach, colon, breast, and thyroid samples. Of the 33 CpG sites examined, 10 non-island-CpG sites, but none of the CpG-island margins were undermethylated concurrent with tissue-specific expression of their nearby genes. The net methylation of the 33 CpG sites and the net amount of non-island-CpG gene transcripts were high in stomach tissues and low in stromal cells. The present findings suggest that the methylation of the non-island-CpG sites is inversely associated with the expression of the nearby genes, and the concert effect of transitional-CpG methylation is linearly associated with the stomach-specific genes lacking CpG-islands.

A Comparative Histomorphometric Study of the Stomach of Rat (Rattus norvegicus), Bat (Eidolon helvum) and Pangolin (Manis tricuspis) in Relation to Diet / Estudio Comparativo Histomorfométrico del Estómago de Rata (Rattus norvegicus), Murciélago (Eidolon helvum) y Pangolin (Manis tricuspis) en Relación con la Dieta

This study verified the comparative histomorphometric adaptations in the stomach of rat, bat and pangolin in relation to diet. Ten rats, ten bats and ten pangolins of both sexes were used for this investigation. The animals were sacrificed after slight anesthesia under chloroform inhalation. The stomach were excised, fixed in 10 percent formol saline and processed for light microscopic study. Stained slides were also subjected to morphometric analysis at a magnification of 400x. The results revealed that the cellular diameter/ density of parietal and zymogenic cells are significantly different in the three mammals (p<0.05) with the exception of the diameter of the zymogenic cells in pangolin which was not statistically significant (p>0.05) when compared with that of rat. Also, histological analysis revealed slight differences in the pattern of organization and distribution of connective tissue fibers. All these observations were reflections of the different pattern the stomachs of the three mammals have adopted to cope with their respective diets.

En este estudio se verificaron las adaptaciones histomorfométricas comparativas en el estómago de ratas, murciélagos y pangolines en relación a la dieta. Se utilizaron para esta investigación 10 ejemplares de cada especie, de ambos sexos. Los animales fueron sacrificados después de anestesia bajo inhalación de cloroformo. Los estómagos fueron extirpados, fijados en formol al 10 por ciento de solución salina y procesados para su estudio microscópico de luz. Los cortes teñidos fueron también objeto de análisis morfométrico con un aumento de X 400. Los resultados revelaron que el diámetro/densidad celular de parietal y las células cimógenas son significativamente diferentes en los tres mamíferos (p <0,05), con la excepción del diámetro de la células cimógenas de pangolines que no era estadísticamente significativa (p> 0,05) en comparación con la de rata. Por otra parte, el análisis histológico reveló ligeras diferencias en las características de organización y distribución de las fibras del tejido conjuntivo. Todas estas observaciones son un reflejo del patrón de los diferentes estómagos de los tres mamíferos, que han adoptado para hacer frente a sus respectivas dietas.

Microarchitectural adaptations in the stomach of African tree pangolin (Manis tricuspis) / Adaptaciones microarquitecturales en el estómago del pangolin africano (Manis tricuspis)

The microarchitecture of the pangolin's stomach favouring the high chitinous diet has been less waived into, despite extensive morphological investigations. Histological analysis of the microanatomy will provide powerful tools for interpretation to yield reliable insights. We investigated this by fixing the tissues in 10 percent formol saline for histological analysis. Serial sections at 5 micron m thickness were subjected to general staining methods for light microscopic study (Haematoxylin and eosin, Van Gieson's and Verhoeff's). The results revealed basic structural arrangements in their coats, with a modification of the epithelial lining of cardia and fundus into stratified squamous keratinized epithelium. These modifications were also reflected in the distribution of collagen and elastic fibers in the various layers (coats) of the stomach. The present study has shown that there was an adaptation of the stomach of African tree pangolin to its diet as reflected in the microarchitectural configuration.

La micro arquitectura del estómago de los pangolines que favorece la alta dieta de chitinous sido poco tomada en cuenta, a pesar de las amplias investigaciones morfológicas. El análisis histológico de la microanatomía proporcionará herramientas de gran importancia para la interpretación, junto con dar una información confiable. Se investigó mediante la fijación de los tejidos en solución salina de formol al 10 por ciento para análisis histológico. Las serie de secciones fueron sometidos a métodos de tinción estándar para el estudio con microscopía de luz (hematoxilina y eosina, Van Gieson y Verhoeff s). Los resultados revelaron adaptaciones estructurales básicas en sus capas, con una modificación del revestimiento epitelial del cardias y fundus en epitelio escamoso estratificado (queratinizado). Estas modificaciones también se reflejan en la distribución de colágeno y fibras elásticas en las diversas capas del estómago. El presente estudio ha demostrado que es una adaptación del estómago a la dieta como se refleja en la configuración de la microarquitectura.

Histological description of the midgut and the pyloric valve of Tropidacris collaris (Stoll, 1813) (Orthopetera: Romaleidae)

The present research describes the histology of the midgut, gastric caeca, and pyloric valve of Tropidacris collaris (Stoll, 1813), (Orthopetera: Romaleidae). We used light microscopy, staining (Gomori's trichrome and periodic acid-Schiff (PAS)), and a routine histological analysis method (hematoxilin-eosin). The insects were obtained from, and also bred in, the Laboratory of Entomology, Department of Biology, of the Rural Federal University of Pernambuco (UFRPE). The collected material was fixed in alcoholic Boüin and embedded in paraplast. The results demonstrated that the midgut wall is composed of an inner epithelial layer and two outer layers of striate muscles: one internal (circular) and the other external (longitudinal), with connective tissue between the muscle fibers. The epithelium is single-layered, with two cell types: regenerative and elongated columnar. The gastric caeca presents muscle layers similar to those of the midgut. Simple columnar epithelium lines the gastric caeca, which presents villi and projects towards the lumen. The pyloric valve is of striate muscle tissue, covered by a single epithelial-cell layer.

A presente pesquisa teve como objetivo descrever a histologia do mesêntero, cecos gástricos e válvula pilórica de Tropidacris collaris (Stoll, 1813) (Orthoptera: Romaleidae), por meio da microscopia de luz, utilizando-se colorações especiais (Tricrômico de Gomori e P.A.S. - Acido periódico de Schiff) e de rotina (Hematoxilina-Eosina). Os insetos foram obtidos da criação existente no Laboratório de Entomologia, do Departamento de Biologia, da UFRPE. O material coletado foi fixado em Boüin alcoólico e processado para inclusão em "paralast". Os resultados mostraram que a parede do mesêntero é constituída por duas camadas de músculo estriado: uma interna (circular) e outra externa (longitudinal), com a presença de tecido conjuntivo entre as fibras musculares. O epitélio é do tipo simples, constituído por dois tipos de células: células regenerativas e células colunares altas. Os cecos gástricos também apresentam as camadas de músculo semelhantes às observadas no mesêntero. O tecido epitelial é do tipo simples colunar projetando-se para a luz do órgão, formando vilosidades. A válvula pilórica é constituída por tecido muscular estriado, recoberto por tecido epitelial.

Tumores del estroma gastrointestinal (GIST): revisión de la literatura / Gastrointestinal stromal tumors (gist): literature review

Los tumores del estroma gastrointestinal representan un tipo infrecuente de tumor de origen mesenquimático, con origen en las células intersticiales de Cajal. Se caracterizan por expresar un receptor de membrana mutante con actividad tirosina quinasa anormal (c-kit CD117) que condiciona su activación permanente y una proliferación celular no controlada. Pueden encontrarse a cualquier nivel del tracto gastrointestinal, pero son más frecuentes en estómago e intestino delgado. Frecuentemente cursan de forma asintomática, constituyendo un hallazgo endoscópico o radiológico. La enfermedad localizada habitualmente es de buen pronóstico tras su resección quirúrgica, mientras que la sobrevida es baja en etapas avanzadas, con escasa respuesta a la quimioterapia convencional. El surgimiento del imatinib, un inhibidor de la tirosina quinasa, ha representado un verdadero hito, dado su eficacia en el control de la enfermedad irresecable o metastásica, permitiendo un importante aumento en la sobrevida de estos pacientes.

Effect of Acupuncture in the Treatment of Young Pigs with Induced Escherichia coli Diarrhea

The effect of acupuncture in the treatment of young pigs with induced enteropathogenic Escherichia coli diarrhea was histopathologically evaluated by routine hematoxylin and eosin stain. Thirty two pigs weighed 4-5kg and aged 21days old were used in this study. The animals with diarrhea were treated with traditional acupuncture, or enrofloxacin. In the group treated with traditional acupuncture, acupoint GV1 (Jiaochao) was used and in the group treated with antibiotics, enrofloxacin was injected intramuscularly. Ten pigs were inoculated with E. coli, but were not treated and served as nontreated control group. At postinoculation day 6, all pigs of the acupuncture and antibiotic treated groups recovered from diarrhea. In the ascending and descending colons of the nontreated control group, severe infiltration of inflammatory cells in the lamina propria was observed and in the fundic stomach, destruction of the fundic gland architecture and necrotic lesions were observed, however, in the same sites of the acupuncture and antibiotics treated groups, the mucosae of the colon and stomach were relatively similar to those of the normal group. These results indicate that acupuncture treatment is effective in controlling induced E. coli diarrhea in pigs at its early stage.

Effects of somatostatin, LHRH and TGF-(alpha) on epithelial cell proliferation in fetal stomach maintained in organ culture

An organ culture technique was used to examine the effects of somatostatin, luteinizing hormone releasing hormone (LHRH) and transforming growth factor (TGF-alpha) on epithelial cell proliferation in rat fetal stomach. The explants were obtained from 20-day rat fetuses and were maintained in organ culture for 24 h. Half of the culture dishes were supplemented with 10 percent fetal bovine serum (FBS). Cell proliferation was assessed using the metaphasic index. Light and electron microscopy showed that the explants could be mainteined in good condition, independent of the FBS or hormone treatment. Re-epithelialization occurred at the edges of the fragments. The addition of 10 percent FBS was not advantageous for evaluation cell proliferation in this organ culture system. The low metaphasic index showed that somatostatin and LHRH significantly inhibited cell proliferation after 24 h of treatment. In contrast, TGF-alpha had a mitogenic effect on fetal gastric mucosa and prevented glandular degeneration. These results corroborate our previous studies in vivo and provide direct evidence of the influence of hormonal and growth factors on gastric mucosa during fetal development.

Tumor del estroma gastrointestinal: informe de un paciente / Gastrointestinal stromal tumor: case report

Se informan los hallazgos sonográficos de un tumor del estroma gastrointestinal, asintomático y que coincidió con vómitos de origen central, en un paciente de 76 años de edad. En este caso, el ultrasonido fue la herramienta diagnóstica más útil, comparándola con otros estudios como la Tomografía Computada (TAC) y la serie Gastroduodenal (SGD). La endoscopia fue un método complementario valioso.

Evaluación de una prueba rápida de ureasa (Hp-INS): Comparación con CU-Test e Histología / Clinical evaluatrion of a new Helicobacter rapid Test (Hp-INS). comparison with CU-Test and Histology

Antecedentes: Las pruebas rápidas de ureasa (PRU) en biopsias gástricas para la detección de Helicobacter pylori (Hp) son prácticas y de fácil aplicación, pero su costo es elevado. Objetivo: Comparar una nueva prueba de ureasa rápida (HP-INS) con una prueba comercial (CU-TEST) e histología en el diagnóstico de Hp. Método: Se obtuvieron cuatro biopsias de antro y dos de cuerpo gástrico en 100 pacientes atendidos consecutivamente en el Servicio de endoscopia entre mayo y julio de 1998. Se destinó una biopsia de antro para cada PRU y las restantes se enviaron a patología. La lectura de las PRU se realizaron simultáneamente a 1, 4 y 24 horas. El patólogo desconocía el resultado de las PRU. Se realizó una encuesta para datos demográficos e historia de fármacos usados en el mes previo al estudio. Resultados: Se analizaron 98 pacientes (58 hombres y 40 mujeres), dos se descartaron. La edad promedio fue de 53.6 años (17-87 años). La endoscopia mostró gastropatía nodular/eritematosa en 75 casos (76 por ciento), úlcera gástrica en nueve (9 por ciento) y duodenal en 9 casos (9 por ciento). El 73 por ciento de los pacientes habían recibido inhibidores de la secreción gástrica y/o antibióticos en las cuatro semanas previas y 42 (43 por ciento) los recibían al momento del estudio. La histología fue positiva en 55 pacientes (56 por ciento, el CU-TEST y el Hp-INS) en 46 casos (47 por ciento) y 40 casos (41 por ciento) respectivamente. La sensibilidad y especificidad para el CU-TEST fue de 71 y 84 por ciento respectivamente y para el Hp-INS de 62 y 80 por ciento respectivamente. Conclusiones: El Hp-INS) es una prueba comparable en igualdad de condiciones al CU-TEST para detectar Hp. Es necesario realizar más estudios excluyendo las variables de confusión para establecer la exactitud diagnóstica de la prueba.

Histology revisited: technical improvement promoted by the use of hydrophilic resin embedding

The use of isotonic 4 per cent paraformaldeyde fixation and glycolmethacrylate embedding permits to obtain undistorted light microscope images with an increased resolution to some extent similar to low power images of electron microscopy. This allows an improved analysis of cell and tissue biology with light microscopy.

Gastric carcinoma in Iraq: evaluation of clinical and diagnostic procedures

Gastric carcinoma is the 8th common and the commonest G.I. cancer in Iraq. 418 patients with biopsy proved cancer were studied clinically and by various diagnostic procedures. Non-specific dyspepsia was the presenting symptom in 2/3 patients and peptic ulcer like pain in 19,7% of patients. Tumor mass in the antrum was the commonest endoscopic finding and an ulcer was found in 23% of patients, but neither size nor site of the ulcer was helpful in differentiating benign from malignant ulcers. Barium series was helpful in 62,% of patients whereas brush cytology was diagnostic in 82% of patients. Histopathology was reported as adenocarcinoma in all but the disease was well advanced in all patients and curative measures were either palliative or not feasible