RESUMO
The purpose of the present study was to examine stimulation of gastrin release and the synthesis of gastrin directly by measurement of incorporation of [(3)H]tryptophan into gastrin in rat antral mucosal explants maintained in organ culture. Gastrin synthesis and secretion were assessed simultaneously at intervals over the 24-h duration of explant culture. Antral mucosal explants from fed female Wistar rats (4-5 wk, 100-150 g) were cultured at 37 degrees C (95% O(2)/5% CO(2)) in medium containing 70% Trowell-T8 and 10% NCTC-135 without unlabeled tryptophan, 10% dialyzed fetal calf serum and [(3)H]tryptophan (100 muCi/ml). Antral tissue was harvested at regular intervals during 24-h culture periods. Incorporation of [(3)H]tryptophan into immunoreactive gastrin was determined by techniques utilizing double-antibody immunoprecipitation. Antral tissue protein synthesis was assessed by measurements of incorporation of [(3)H]tryptophan into tissue protein of cultured antral explants. In paired experiments, gastrin synthesis and secretion in the presence of dibutyryl cAMP (DBCAMP) were compared to those observed under control conditions. Gastrin and protein specific activity progressively increased with time. Gastrin specific activity at 30 min increased from 3.3+/-0.5 (SEM) to 55.2+/-10.6 fmol [(3)H]tryptophan/pmol gastrin (or from 1.57+/-0.48 to 26.28+/-5.05 pmol [(3)H]tryptophan/mug gastrin) at 24 h: specific activity of antral tissue protein at 30 min increased from 33.6+/-8.4 to 1,660+/-236 fmol [(3)H]tryptophan/mug protein at 16 h. Culturing of explants for 4 h in the presence of cycloheximide (100 mug/ml) inhibited both gastrin synthesis and protein synthesis by greater than 90 and 95%, respectively. DBCAMP (10 mM) significantly increased both the synthesis and secretion of antral gastrin when compared with control cultured explants. Results of these experiments provide direct demonstration of gastrin synthesis by rat antral mucosal explants in organ culture, indicate that both gastrin and total antral protein synthesis are inhibited by cycloheximide, and demonstrate DBCAMP-induced stimulation of both gastrin synthesis and secretion, suggesting the potentially important role of cyclic AMP in gastrin cell function.
Assuntos
Bucladesina/farmacologia , Cicloeximida/farmacologia , Gastrinas/fisiologia , Antro Pilórico/fisiologia , Animais , Feminino , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/fisiologia , Gastrinas/biossíntese , Gastrinas/metabolismo , Humanos , Técnicas de Cultura de Órgãos , Biossíntese de Proteínas , Antro Pilórico/efeitos dos fármacos , Antro Pilórico/metabolismo , Ratos , Triptofano/metabolismoRESUMO
Sclerosing peritonitis developed in a 43-year-old man with angina pectoris who had been receiving the beta-adrenergic receptor antagonist, propranolol. The patient had abdominal and back pain, weight loss, a midabdominal fullness, ascites, and evidence of partial small bowel obstruction. At surgery, the small bowel was distended and encased by dense fibrous tissue. Infectious and neoplastic causes of fibrosing peritoneal inflammation were excluded. The patient described in this report illustrates several features commonly experienced by individuals who developed sclerosing peritonitis associated with beta-adrenergic receptor blockade therapy. To my knowledge, the development of ascites and considerable ascitic fluid leukocytosis have not been described previously with this disorder.
Assuntos
Toxidermias/etiologia , Peritonite/induzido quimicamente , Propranolol/efeitos adversos , Adulto , Conjuntivite/induzido quimicamente , Humanos , Masculino , Peritonite/diagnóstico , EscleroseRESUMO
To identify the factors regulating the proliferation of intestinal epithelium, we examined the effects of various growth factors on [3H] thymidine incorporation into the DNA of IEC-6 cells, an intestinal epithelial cell line derived from rat jejunal crypts. Insulin-like growth factor-I (IGF-I), IGF-II, and insulin stimulated the DNA and protein synthesis of IEC-6 cells in serum-free medium supplemented with transferrin, dexamethasone, and BSA (basal medium). Concentration-response experiments demonstrated that IGF-I is approximately 10 times more potent than IGF-II or insulin in producing 2- to 3-fold stimulations of DNA and protein synthesis by IEC-6 cells. In addition, IEC-6 cells proliferated slowly in the basal medium without any added growth factors. Analysis of medium conditioned by IEC-6 cells by gel filtration chromatography, RIA, HPLC, and N-terminal sequencing revealed that IEC-6 cells synthesize and secrete mature, 7,500 mo wt (M(r)) IGF-II as well as high M(r) forms of IGF-II. In addition, ligand blot, immunoblot, and N-terminal sequence analyses showed that IEC-6 cells produce the 34,000 M(r) IGF-binding protein-2 (IGFBP-2). To determine if IGFBP-2 modulates IGF responses in IEC-6 cells, the IGF-I analogs, Des-(1-3)-IGF-I and [Gln3,Ala4,Tyr15,Leu16]IGF-I, both of which have a reduced affinity for IGFBPs, were tested for their effects on IEC-6 cell proliferation. Both analogs exhibited 10-fold greater potency than IGF-I, presumably because endogenously secreted IGFBPs depress IGF-I binding to cell surface receptors. Finally, purified IGFBP-2 attenuated the DNA synthesis of IEC-6 cells in a dose-dependent manner. We conclude that IGFBP-2 secreted by intestinal epithelial cells is capable of limiting the mitogenic activity of both exogenous and endogenous IGFs by blocking the association of the growth factors with cell surface binding sites. These results further suggest that the growth of IEC-6 cells is modulated by autocrine mechanisms involving IGF-II and IGFBP-2.
Assuntos
Proteínas de Transporte/metabolismo , Substâncias de Crescimento/farmacologia , Fator de Crescimento Insulin-Like II/metabolismo , Sequência de Aminoácidos , Análise de Variância , Animais , Proteínas de Transporte/farmacologia , Divisão Celular/efeitos dos fármacos , Linhagem Celular , Meios de Cultura Livres de Soro , DNA/biossíntese , Replicação do DNA/efeitos dos fármacos , Epitélio , Insulina/farmacologia , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina , Fator de Crescimento Insulin-Like I/análogos & derivados , Fator de Crescimento Insulin-Like I/farmacologia , Jejuno , Cinética , Dados de Sequência Molecular , Biossíntese de Proteínas , Ratos , Proteínas Recombinantes/farmacologia , Fator de Crescimento Transformador beta/farmacologiaRESUMO
Hypergastrinemia and gastric acid hypersecretion are the principal laboratory features of Zollinger-Ellison syndrome. Decision and cost-effectiveness analyses were employed in the present study to compare and contrast the diagnostic strategies of initial gastric analysis followed by secretin infusion test versus secretin infusion test alone in the evaluation of hypergastrinemia in patients suspected of having gastrinoma. The results of this study showed that 59 percent of patients with elevated serum gastrin values were either hypochlorhydric or achlorhydric. Application of decision analysis to either diagnostic strategy demonstrated that gastric analysis followed by secretin infusion test, if indicated, was superior in expected value than secretin infusion test alone. Likewise, in this group of patients, performance of gastric analysis in the outpatient setting prior to secretin infusion testing was financially more advantageous than performance of secretin infusion testing alone. These results also demonstrate the importance of performing gastric analysis prior to anticipated hospitalization for evaluation of suspected gastrinoma. Such testing would obviate unnecessary hospitalization and medical costs.
Assuntos
Ácido Gástrico/metabolismo , Síndrome de Zollinger-Ellison/diagnóstico , Adolescente , Adulto , Idoso , Análise Custo-Benefício , Tomada de Decisões , Feminino , Determinação da Acidez Gástrica , Gastrinas/sangue , Humanos , Masculino , Pessoa de Meia-Idade , Radioimunoensaio , SecretinaRESUMO
Sequences encoding the UL1 gene of equine herpesvirus type 1 (EHV-1) are conserved in the genome of EHV-1 defective interfering particles (DIPs) that mediate oncogenic transformation and persistent infection. The UL1 protein was identified by in vitro transcription/translation and hybrid-arrest translation analyses which employed a UL1/pGEM-3Z construct designated pGEML1. SDS-PAGE analyses of in vitro translation products synthesized from UL1-specific RNA revealed that the UL1 ORF encodes a 30 kDa protein which corresponds in size to the 258 amino acid protein predicted by DNA sequence analyses. This result was confirmed by arresting translation of the in vitro transcribed UL1 RNA with an oligodeoxynucleotide complementary to UL1 coding sequences. The UL1 protein is a homolog of the predicted protein encoded by the ORF2 gene of varicella-zoster virus, but UL1 has no homolog in herpes simplex virus type 1. The UL1 protein contains a region conforming to a 'PEST' (Proline, Glutamic acid, Serine, and Threonine) sequence, which is commonly found in proteins with half-lives of less than two hours.
Assuntos
Herpesvirus Equídeo 1/genética , Proteínas Virais/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA Viral/genética , Expressão Gênica , Genes Virais , Dados de Sequência Molecular , Fases de Leitura Aberta , Biossíntese de Proteínas , RNA Viral/genética , Transcrição GênicaRESUMO
The effects of alterations in availability and access of extracellular media calcium on antral gastrin release were examined in the basal state and in response to cholinergic stimulation in rat antral organ culture experiments. In the presence of either divalent cationic chelator (EGTA) or calcium channel blocker (verapamil, nifedipine), carbachol-stimulated gastrin release was inhibited completely to values that were not significantly different from non-stimulated control. In the absence of added calcium chloride, carbachol stimulated gastrin release during the initial 30 min of culture but not at 69 and 120 min of culture. Inhibition by EGTA and verapamil of carbachol-stimulated gastrin release during the initial 30 min of culture suggests, but does not prove, that these agents may also effect intracellular availability and movement of calcium. Cholinergic stimulation of gastrin release demonstrated a concentration-dependent relationship with extracellular calcium: optimal culture media calcium concentration was 1 mM. In conclusion, these studies indicate that cholinergic stimulation of the gastrin cell requires availability of extracellular calcium.
Assuntos
Cálcio/fisiologia , Carbacol/farmacologia , Gastrinas/metabolismo , Animais , Ácido Egtázico/farmacologia , Técnicas In Vitro , Masculino , Nifedipino/farmacologia , Ratos , Ratos Endogâmicos , Triptofano/metabolismo , Verapamil/farmacologiaRESUMO
Growth of neomucosa has been investigated as a means to increase intestinal surface area in the short-bowel syndrome. Functional neomucosa grows over patched intestinal defects, but the effect of the patching procedure on absorption is unknown. The purpose of this study was to determine morphologic and nutritional responses to intestinal patching after resection. Fifteen dogs (13 to 19 kg) underwent either 75% resection of the small intestine (control group, n = 5), simultaneous resection and patching of the intestinal remnant with colon serosa (simultaneous group, n = 5), or resection with patching 12 weeks later (delayed group, n = 5). Caloric intake was standard in the three groups. Animals were killed 40 weeks after resection or patching. At that time, defects were 95% covered with neomucosa in both patched groups. Intestinal remnant length increased significantly in controls (139 +/- 20% initial length) compared to the simultaneous group (99 +/- 6%, p less than 0.05) but not to the delayed group (119 +/- 11%). Villous height of intestinal mucosa was greater in the control and delayed groups than in the simultaneous group (714 +/- 36 and 624 +/- 111 versus 535 +/- 54 micron, p less than 0.05). Fasting gastrin levels were significantly greater in patched animals than after resection alone (p less than 0.05). Intestinal transit by barium meal was significantly longer in patched animals (18 +/- 7 minutes versus 11 +/- 6, p less than 0.05). Body weight and serum albumin level were significantly lower in patched animals at death. Fecal weight, moisture, and fat excretion were significantly increased in the simultaneous group. Although intestinal patching results in the growth of neomucosa and prolonged transit time, it has a deleterious effect on absorption and nutritional status. In part, this may be related to inhibition of intestinal adaptation and gastric hypersecretion in patched animals.
Assuntos
Mucosa Intestinal/citologia , Intestinos/cirurgia , Estado Nutricional , Animais , Peso Corporal , Colesterol/sangue , Colo/cirurgia , Cães , Fezes/análise , Gastrinas/sangue , Trânsito Gastrointestinal , Íleo/cirurgia , Absorção Intestinal , Mucosa Intestinal/crescimento & desenvolvimento , Intestinos/citologia , Albumina Sérica/análiseRESUMO
Somatostatin analogue octreotide inhibits intestinal absorption and motility but its effect on epithelial cell migration and proliferation remains unclear. Our aim was to determine the effect of octreotide on parameters of intestinal regeneration, including epidermal growth factor (EGF)-induced changes. Thirty rabbits had full-thickness ileal defects patched with cecal serosa surface. Group 1 were controls. Groups 2 and 3 received 100 micrograms and 1000 micrograms, respectively, of subcutaneous octreotide daily. Group 4 received EGF at 1.5 micrograms/kg per hour via subcutaneous miniosmotic pump, and group 5 received both octreotide (1000 micrograms/d) and EGF (1.5 micrograms/kg per hour). Octreotide at 100 micrograms/d did not inhibit epithelial cell migration or proliferation at 7 days. Octreotide at 1000 micrograms/d inhibited normal but not EGF-stimulated cell migration. Octreotide decreased EGF-stimulated but not normal proliferation. Octreotide impairs epithelial cell migration in a dose-dependent manner. Octreotide inhibits EGF-stimulated proliferative activity but not EGF-stimulated migration. Prolonged administration of octreotide may adversely affect normal and adaptive intestinal regeneration by both direct and indirect effects.
Assuntos
Intestinos/fisiologia , Octreotida/farmacologia , Regeneração/efeitos dos fármacos , Animais , Ceco/transplante , Movimento Celular/efeitos dos fármacos , Dissacaridases/metabolismo , Fator de Crescimento Epidérmico/farmacologia , Epitélio/efeitos dos fármacos , Epitélio/fisiologia , Gastrinas/sangue , Íleo/fisiologia , Íleo/cirurgia , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/enzimologia , Intestinos/efeitos dos fármacos , Masculino , Coelhos , Proteínas Recombinantes , Sacarase/metabolismoRESUMO
Rat antral mucosal fragments were maintained in short-term culture to examine the relative potencies and receptor specificity of the cholinergic agonist, carbachol, and adrenergic agents, norepinephrine, isoproterenol, clonidine and phenylephrine in stimulating gastrin release. Results of these studies indicate that norepinephrine and carbachol evoke pharmacologically and temporally distinctive patterns of antral gastrin release. Dose-response experiments indicate that norepinephrine is approximately 10,000 times more potent on a molar basis than carbachol in stimulating antral gastrin release. Adrenergic (norepinephrine, isoproterenol) stimulation of antral gastrin release was prevented by propranolol, and cholinergic- (carbachol) mediated peptide release was blocked by both atropine and pirenzepine. Phenylephrine and clonidine did not alter basal gastrin release. The pattern of peptide release as a function of time was quite different for each agent: norepinephrine exerted its stimulatory effect acutely during the initial 30 minutes of incubation, while carbachol exhibited a sustained stimulatory action throughout the 2-hour culture period. In conclusion, data from these studies suggests that there are marked differences between norepinephrine and carbachol in their pharmacological potency and time-dependent activation of the G cell.
Assuntos
Carbacol/farmacologia , Mucosa Gástrica/metabolismo , Gastrinas/metabolismo , Norepinefrina/farmacologia , Antro Pilórico/metabolismo , Receptores Adrenérgicos/fisiologia , Receptores Colinérgicos/fisiologia , Animais , Atropina/farmacologia , Clonidina/farmacologia , Mucosa Gástrica/efeitos dos fármacos , Masculino , Técnicas de Cultura de Órgãos , Fenilefrina/farmacologia , Pirenzepina/farmacologia , Antro Pilórico/efeitos dos fármacos , Ratos , Ratos Endogâmicos , Ioimbina/farmacologiaRESUMO
The effects of the microtubule stabilizing agent, deuterium oxide, on in vitro rat antral gastrin release were examined under basal conditions and during stimulation with isobutyl methylxanthine and bombesin plus isobutyl methylxanthine. Basal gastrin release from antral mucosal fragments was unaffected by increasing media concentration of deuterium oxide (12.5 to 75% v/v) during 1 h incubations. Gastrin release stimulated by isobutyl methylxanthine (0.1 mM), a potent inhibitor of phosphodiesterase activity, was inhibited completely by 12.5% deuterium oxide. Bombesin (1 X 10(-8) M) in the presence of IBMX (0.1 mM) stimulated gastrin release (29.7 +/- 1.9% of total gastrin). This was significantly greater than gastrin released under control conditions and with IBMX alone: 12.0 +/- 1.1 (P less than 0.001) and 20.2 +/- 2.6% of total gastrin (P less than 0.02), respectively. Partial inhibition of bombesin-IBMX stimulated gastrin release was achieved with 12.5% and 25% deuterium oxide and stimulation of gastrin release was inhibited completely by 50% deuterium oxide. In contrast to these results, gastrin release stimulated by the calcium ionophore A23187 was not inhibited by 50% deuterium oxide. Additional studies were performed to assess reversibility of the effects of deuterium oxide on stimulated gastrin release. Antral tissue exposed to initial culture medium containing deuterium oxide (50%) and bombesin-IBMX for 60 min was exchanged for medium without deuterium oxide. Restimulation of antral tissue during the second hour of culture resulted in gastrin release that was comparable to that observed in cultures not exposed to deuterium oxide during the first hour of culture. Reversibility of the effects of deuterium oxide suggest that a functional alteration in microtubular function is restored by removal of heavy water from the culture medium. Results of these experiments indicate that deuterium oxide is capable of inhibiting gastrin release stimulated by the peptide hormone bombesin and by the phosphodiesterase inhibitor isobutyl methylxanthine. Furthermore, these results suggest that increased levels of intracellular calcium achieved by the action of ionophore A23187 prevent microtubular stabilization by deuterium oxide.
Assuntos
Deutério/farmacologia , Mucosa Gástrica/metabolismo , Gastrinas/metabolismo , Antro Pilórico/metabolismo , Água/farmacologia , 1-Metil-3-Isobutilxantina/farmacologia , Animais , Bombesina/farmacologia , Calcimicina/farmacologia , Óxido de Deutério , Relação Dose-Resposta a Droga , Mucosa Gástrica/efeitos dos fármacos , Técnicas In Vitro , Cinética , Masculino , Antro Pilórico/efeitos dos fármacos , Ratos , Ratos EndogâmicosRESUMO
Anorexia nervosa (AN) is a syndrome of unknown cause characterized by voluntary starvation. Cholecystokinin has been implicated as a neuroendocrine regulatory factor in control of satiety. Relatively little information is known about gastrointestinal hormone responses to feeding in subjects with anorexia nervosa. In the present studies, we examine fasting and postprandial levels of cholecystokinin (CCK), vasoactive intestinal peptide (VIP) and peptide histidine methionine (PHM) in anorexia nervosa subjects and in control individuals. Results of these studies indicate that plasma CCK response to a liquid meal (Ensure Plus) in untreated AN subjects was distinctly different from that observed in healthy controls, both in terms of temporal pattern of peptide released and the amount of CCK secreted into the circulation. Peak levels of CCK release occurred at 30 min following meal ingestion in AN patients and at 60 min in control subjects. Integrated CCK release in untreated AN patients was approximately twice that measured in control individuals. Renutrition therapy was associated with reversion of the pattern of CCK release to that observed in control subjects. Plasma VIP levels were unchanged following meal ingestion in both control and anorexic subjects. In contrast, PHM levels in AN subjects were significantly greater than that observed in control individuals. The pattern of PHM release following liquid meal ingestion was similar to that observed with plasma CCK; namely, peak release of peptide was observed at 30 min which was significantly greater than corresponding control values (P less than 0.05). In conclusion, these results demonstrate distinctive differences in plasma CCK and PHM levels in response to feeding in AN subjects when compared to control individuals. These findings suggest that the earlier and greater rise in plasma CCK levels in AN subjects following meal ingestion may contribute to the abnormal sensation of satiety in this condition.
Assuntos
Anorexia Nervosa/etiologia , Dietoterapia , Ingestão de Alimentos/fisiologia , Hormônios Gastrointestinais/metabolismo , Peptídeo PHI/metabolismo , Anorexia Nervosa/terapia , Colecistocinina/metabolismo , Feminino , Alimentos Formulados , Humanos , Plasma/química , Peptídeo Intestinal Vasoativo/metabolismoRESUMO
The ability of exogenous calcitonin gene-related peptide (CGRP) to regulate gastric somatostatin and gastrin messenger RNA was studied in vitro in rat antral mucosal/submucosal tissues. Somatostatin and gastrin mRNA were quantified by Northern and dot blot hybridization and regulatory peptides were measured by radioimmunoassay. Incubation of antral tissues in the presence of CGRP [1 x 10(-7) M] for 60 min resulted in a reciprocal increase in somatostatin and a decrease in gastrin release: 214.7+/-28.5 vs. control of 81.7+/-5.9 pg somatostatin per gram of tissue and 2.2+/-0.3 vs. control of 5.5+/-0.7 ng gastrin per gram of tissue (P < 0.001). CGRP caused parallel changes in somatostatin and gastrin mRNA levels: somatostatin mRNA increased by 212% from 0.40+/-0.02 to 1.25+/-0.09 absorbance units (AU) (P < 0.001) and gastrin mRNA decreased by 73% from 0.55+/-0.08 to 0.15+/-0.02 AU (P < 0.001). Somatostatin monoclonal antibody prevented CGRP-mediated inhibition of both gastrin release and gastrin mRNA levels. In conclusion, CGRP is capable of modulating both the secretion and gene expression of regulatory peptides from antral G and D cells. Somatostatin immunoneutralization studies suggest that the actions of CGRP on gastrin release and gene expression are indirect and mediated through the paracrine influences of somatostatin.
Assuntos
Peptídeo Relacionado com Gene de Calcitonina/farmacologia , Gastrinas/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Antro Pilórico/efeitos dos fármacos , Somatostatina/genética , Animais , Northern Blotting , Western Blotting , Cinética , Masculino , Antro Pilórico/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
Mechanisms of acid-evoked CGRP release from gastric afferent nerves were investigated in rat antral mucosal/submucosal tissues. Low pH (pH 4.0, 5.0 and 6.0) stimulated antral CGRP release significantly and dose-dependently from rat antral fragments. Removal of extracellular calcium from the incubation medium resulted in significant inhibition (59%, P < 0.001) of acid (pH 4.0)-stimulated CGRP release. Conotoxin (1 x 10(-7) M), the selective blocker of N-type calcium channels, also significantly inhibited proton (pH 4.0)-induced CGRP release to values that were 74% below net stimulated levels. Neither nifedipine (1 x 10(-6) M), the L-type Ca(2+)-channel antagonist, nor indomethacin (1 x 10(-5) M), inhibitor of prostaglandin synthesis, altered acid-induced CGRP release. In contrast, ruthenium red (1 x 10(-5) M), capsaicin antagonist, almost completely prevented acid (pH 4.0)-stimulated CGRP release. Capsazepine (1 x 10(-4) M), a specific capsaicin receptor antagonist, also completely abolished acid-induced CGRP release. In conclusion, the results of these studies indicate that hydrogen ions are capable of evoking CGRP release from peripheral sensory neurons in rat antral mucosal/submucosal tissues. Proton-evoked CGRP release requires extracellular calcium and involves N-type calcium channels. Furthermore, acid appears to exert a capsaicin-like effect to evoke sensory neuropeptide release that is sensitive to capsazepine and ruthenium red. These data suggest that proton-induced antral CGRP release represents a direct action of hydrogen ions on mucosal/submucosal sensory dendritic nerve endings to effect local release of neuropeptide.
Assuntos
Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Concentração de Íons de Hidrogênio , Antro Pilórico/metabolismo , Animais , Toxina da Cólera/farmacologia , Indometacina/farmacologia , Masculino , Nifedipino/farmacologia , Antro Pilórico/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Tetrodotoxina/farmacologia , Fatores de TempoRESUMO
Outer membrane protein F of Pseudomonas aeruginosa has vaccine efficacy against infection by P. aeruginosa as demonstrated in a variety of animal models. Through the use of synthetic peptides, three surface-exposed epitopes have been identified. These are called peptides 9 (aa 261-274 in the mature F protein, TDAYNQKLSERRAN), 10 (aa 305-318, NATAEGRAINRRVE), and 18 (aa 282-295, NEYGVEGGRVNAVG). Both the peptide 9 and 10 epitopes are protective when administered as a vaccine. In order to develop a vaccine that is suitable for use in humans, including infants with cystic fibrosis, the use of viral vector systems to present the protective epitopes has been investigated. An 11-amino acid portion of epitope 10 (AEGRAINRRVE) was successfully inserted into the antigenic B site of the hemagglutinin on the surface of influenza virus. This chimeric influenza virus protects against challenge with P. aeruginosa in the mouse model of chronic pulmonary infection. Attempts to derive a chimeric influenza virus carrying epitope 9 have been unsuccessful. A chimeric plant virus, cowpea mosaic virus (CPMV), with epitopes 18 and 10 expressed in tandem on the large coat protein subunit (CPMV-PAE5) was found to elicit antibodies that reacted exclusively with the 10 epitope and not with epitope 18. Use of this chimeric virus as a vaccine afforded protection against challenge with P. aeruginosa in the mouse model of chronic pulmonary infection. Chimeric CPMVs with a single peptide containing epitopes 9 and 18 expressed on either of the coat proteins are in the process of being evaluated. Epitope 9 was successfully expressed on the coat protein of tobacco mosaic virus (TMV), and this chimeric virus is protective when used as a vaccine in the mouse model of chronic pulmonary infection. However, initial attempts to express epitope 10 on the coat protein of TMV have been unsuccessful. Efforts are continuing to construct chimeric viruses that express both the 9 and 10 epitopes in the same virus vector system. Ideally, the use of a vaccine containing two epitopes of protein F is desirable in order to greatly reduce the likelihood of selecting a variant of P. aeruginosa that escapes protective antibodies in immunized humans via a mutation in a single epitope within protein F. When the chimeric influenza virus containing epitope 10 and the chimeric TMV containing epitope 9 were given together as a combined vaccine, the immunized mice produced antibodies directed toward both epitopes 9 and 10. The combined vaccine afforded protection against challenge with P. aeruginosa in the chronic pulmonary infection model at approximately the same level of efficacy as provided by the individual chimeric virus vaccines. These results prove in principle that a combined chimeric viral vaccine presenting both epitopes 9 and 10 of protein F has vaccine potential warranting continued development into a vaccine for use in humans.
Assuntos
Vacinas Bacterianas/imunologia , Vírus da Influenza A/genética , Pneumopatias/prevenção & controle , Vírus de Plantas/genética , Porinas/imunologia , Infecções por Pseudomonas/prevenção & controle , Pseudomonas aeruginosa/imunologia , Animais , Anticorpos Antibacterianos/sangue , Vacinas Bacterianas/administração & dosagem , Comovirus/genética , Comovirus/metabolismo , Ensaio de Imunoadsorção Enzimática , Epitopos/genética , Epitopos/imunologia , Epitopos/metabolismo , Vírus da Influenza A/metabolismo , Pulmão/microbiologia , Pneumopatias/microbiologia , Camundongos , Vírus de Plantas/metabolismo , Porinas/química , Porinas/metabolismo , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/metabolismo , Vírus do Mosaico do Tabaco/genética , Vírus do Mosaico do Tabaco/metabolismo , Vacinação , Vacinas Combinadas/administração & dosagem , Vacinas Combinadas/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/imunologiaRESUMO
Immune disorders of the gastrointestinal tract and hepatobiliary systems comprise a diverse group of illnesses which share in common certain overlapping and yet distinctive expressions of cellular and humoral immunity. As is evident from material contained in this article, controversy and disparate results frequently characterize the study of immune mechanisms in a given disease process. Nonetheless, advances in quantitation of specific immunocyte function and phenotypic expression have greatly facilitated the depth of understanding of the immune process related to these disorders. Challenges for future clinical investigation of these disorders are to characterize cell-specific target antigens to which immunologic attack is directed and to unravel the immunogenetic mechanisms that trigger and direct immune-mediated injury to host tissues. It is anticipated that continued investigation of immune disorders of the gastrointestinal tract and liver will clarify pathogenetic mechanisms and thus permit formulation of rational and effective therapies.
Assuntos
Gastroenteropatias/imunologia , Hepatopatias/imunologia , Anemia Perniciosa/etiologia , Anemia Perniciosa/imunologia , Doenças Autoimunes/imunologia , Doença Celíaca/imunologia , Doença Crônica , Doenças Funcionais do Colo/imunologia , Gastrite/classificação , Gastrite/etiologia , Gastrite/imunologia , Hepatite B/imunologia , Hepatite B/patologia , Hepatite Crônica/imunologia , Humanos , Mucosa Intestinal/citologia , Mucosa Intestinal/imunologia , Fígado/patologia , Cirrose Hepática Biliar/imunologia , Cirrose Hepática Biliar/patologia , Hepatopatias/patologia , Testes SorológicosRESUMO
In the present study we developed an experimental model for direct assessment of antral endocrine cell and cholinergic neural responses to luminal stimulation. A sleeve of antral mucosal/submucosal tissue was prepared from rat antrum, mounted in perfusion chamber, and perfused in both luminal and submucosal compartments. Morphological and functional integrity of the antral sleeve were confirmed by histological examination and measurement of protein synthesis. Antral gastrin release was assessed in response to luminal stimulation with acid, peptone and distension. Luminal acid (pH3) inhibited basal gastrin release by -70.4% and luminal peptone stimulated gastrin release to 210% above control (p < 0.02). Distention of the antral sleeve by hydrostatic pressure (3-25cm H2O) caused stepwise and significant increase in gastrin release that was reversible. 3H-acetylcholine was stimulated significantly by KCl (56mM) to values twice control. In summary, these results establish the integrity and responsiveness of the antral sleeve to pharmacological and luminal stimulation. The antral sleeve may be a useful model in assessing antral function in response to luminal stimulation.
Assuntos
Acetilcolina/metabolismo , Gastrinas/metabolismo , Antro Pilórico/metabolismo , Animais , Carbacol/farmacologia , Ácido Gástrico/fisiologia , Masculino , Peptonas/farmacologia , Estimulação Física , Cloreto de Potássio/farmacologia , Pressão , Antro Pilórico/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Estimulação QuímicaRESUMO
A model was designed to permit sampling of portal venous blood in nonanesthetized dogs. Five mongrel dogs were prepared with indwelling portal venous catheters and gastric fistulas. After a minimum recovery period of 7 days, a 10 percent peptone meal was infused into the stomach. Portal and peripheral venous sera were obtained in the basal state and at various time points after intragastric peptone infusion. Serum gastrin concentrations were later measured by radioimmunoassay. Mean basal gastrin from portal and peripheral venous sera were 17 +/- 5 and 13 +/- 2 pg/ml, respectively. After peptone infusion, a biphasic gastrin response was observed. The first peak in portal venous serum occurred at 2 minutes, increasing to 49 +/- 20 pg/ml, while in the peripheral venous serum, the initial peak occurred at 4 minutes to 33 +/- 4 pg/ml. The second peak component occurred at 30 minutes in both circulations, increasing to 35 +/- 14 pg/ml in portal venous serum and to 29 +/- 8 pg/ml in peripheral venous serum. The results of this study (1) demonstrate that the release of gastrin after a peptone meal in the dog is prompt and biphasic, and (2) provide an effective method for obtaining simultaneous portal and peripheral venous blood for measurement of gastrointestinal peptides in alert, nonanesthesized dogs.
Assuntos
Gastrinas/sangue , Peptonas , Veia Porta , Animais , Cateteres de Demora , Sistema Digestório/irrigação sanguínea , Cães , Mucosa Gástrica/metabolismo , Gastrinas/metabolismo , Modelos Biológicos , RadioimunoensaioRESUMO
It is now evident that hypersecretion of gastric hydrochloric acid is an important pathogenetic element among a variety of heterogeneous factors responsible for the production of common duodenal ulcer. Hypersecretion of gastric acid due to usually strikingly increased circulating levels of gastrin released from gastrinoma tissue is characteristics of patients with to Zollinger-Ellison Syndrome. In contrast, fasting serum gastrin levels are normal in patients with common duodenal ulcer. The polypeptide hormone, gastrin does, however, appear to play subtle and multiple roles in enhancement of gastric acid secretion in duodenal ulcer. Recent evidence suggests that abnormalities in gastrin release and action may be influenced by participation of somatostatin. The hypothesis is proposed for consideration and for further investigation that the multiple subtle abnormalities in gastrin release and parietal cell sensitivity to gastrin may be due to disturbances in the actions or concentrations of locally acting polypeptides, substances which are capable of suppressing gastrin release and its effects (somatostatin), or alternatively, are capable of stimulating release of gastrin into the circulation (bombesin).
Assuntos
Úlcera Duodenal/etiologia , Ácido Gástrico/metabolismo , Gastrinas/biossíntese , Bombesina/biossíntese , Úlcera Duodenal/metabolismo , Humanos , Somatostatina/biossíntese , Síndrome de Zollinger-Ellison/etiologia , Síndrome de Zollinger-Ellison/metabolismoRESUMO
The results of single contrast barium enema were retrospectively correlated with colonoscopically diagnosed colorectal disease in 54 patients (75 lesions). Altogether 66 lesions (88%) were correctly diagnosed. The sensitivity of barium enema for polyps was 81% (26/32). There were three perceptive errors and three polyps 5 mm or less in size were not demonstrated by barium enema. Twenty-nine cases of inflammatory disorders were all correctly diagnosed. One of 12 malignancies was missed by perceptive error. In two cases with vascular malformations the barium enema was normal. 4/9 (44%) of missed lesions were perceptive errors and could have been probably avoided by a second independent reading of films.
Assuntos
Sulfato de Bário , Doenças do Colo/diagnóstico , Colonoscopia , Enema , Doenças Retais/diagnóstico , Doenças do Colo/diagnóstico por imagem , Doenças do Colo/patologia , Humanos , Radiografia , Doenças Retais/diagnóstico por imagem , Doenças Retais/patologiaRESUMO
Massive intestinal resection is associated with transient hypergastrinemia and gastric hypersecretion. Gastric hypersecretion impairs intestinal absorption, but gastrin may be trophic during intestinal adaptation. Our aim was to determine if postresection hypergastrinemia correlates with malabsorption or adaptation. Ten dogs (13 to 19 kg) underwent 75% proximal intestinal resection. Intestinal remnant length and villus height was assessed at 12 weeks (n = 5) and 40 weeks (n = 5). Body weight and serum albumin, as well as stool fat, moisture, and weight, were measured preoperatively and at 4-week intervals for 40 weeks. Fasting serum gastrin values were measured by radioimmunoassay at similar intervals. Significant hypergastrinemia occurred between 4 and 28 weeks postresection. Hypergastrinemia did not correlate with increased intestinal remnant length (r = -.486, p = .407) or villus height (r = -.410, p = .584). Duration of hypergastrinemia (> 100 pg/ml) correlated with percentage of fecal fat at 12 weeks (r = .807, p = .015) and stool weight at 40 weeks (r = .881, p = .046). Thus, postresection hypergastrinemia correlates with early fat malabsorption and increased stool weight, but there is no correlation between hypergastrinemia and adaptation. These findings suggest that gastric hypersecretion, not hypergastrinemia, may be the more important pathophysiologic event after intestinal resection.