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Biomedical subjects

C M Townsend

Publications and source records attributed to C M Townsend.

At least 55 records · Page 3Linked to original sources
PubMed1996

Glycine-extended gastrin potentiates gastrin-stimulated gastric acid secretion in rats.

The purpose of this study was to examine whether an intermediate form of amidated gastrin, glycine-extended gastrin (Gly-G), can stimulate gastric acid secretion in conscious rats prepared with gastric fistulas. Intravenous administration of Gly-G (20 nmol.kg-1.h-1) alone for 2 h did not stimulate gastric acid secretion; however, administration of Gly-G (20 nmol.kg-1.h-1) in combination with a bolus administration of gastrin (9.5 nmol/kg) potentiated acid secretion significantly. Gastric acid secretion in response to gastrin alone and gastrin plus Gly-G (2 nmol.kg-1.h-1) was 109.1 +/- 21.6 and 170.1 +/- 27.7 mueq.kg-1.h-1, respectively (P < 0.05). Gastric acid secretion in response to gastrin alone and gastrin plus Gly-G (20 nmol.kg-1.h-1) was 84.8 +/- 17.5 and 164.1 +/- 29.3 mueq.kg-1.h-1, respectively (P < 0.05). Intravenous administration of Gly-G (20 nmol.kg-1.h-1) failed to increase histamine (1 mg/kg)-stimulated acid output. These results demonstrate that Gly-G can selectively potentiate the stimulatory effect of gastrin on acid secretion in rats and that the unprocessed form of gastrin, Gly-G, can exert a biological effect in the stomach.

Animals↗
PubMed1996

Regulation of peptide YY homeostasis by gastric acid and gastrin.

Peptide YY (PYY) is a gut hormone localized primarily in the distal bowel. Because circulating PYY inhibits gastric acid secretion, we investigated the effects of gastric acid secretion and gastrin on gene expression and secretion of PYY. In conscious dogs, PYY release in response to oral food was inhibited (P < 0.05) by pharmacologic inhibition of gastric acid secretion (omeprazole, famotidine). In rats, omeprazole treatment resulted in a significant elevation in serum gastrin concentrations and a simultaneous decrease in PYY messenger RNA (mRNA) and peptide levels in the colon; administration of a gastrin receptor antagonist (L365, 260) prevented the inhibitory actions of omeprazole on colonic PYY mRNA levels. In athymic-nude mice, implantation of a human gastrinoma resulted in an elevation of serum gastrin concentrations and a concomitant depression of colonic PYY mRNA levels. We conclude that endogenous gastric acid secretion up-regulates PYY release and PYY mRNA expression. Circulating gastrin acts to down-regulate PYY release and PYY mRNA expression. This study provides evidence that foregut functions (i.e., gastric acid secretion and gastrin release) exert control over an antiacid signal (e.g. PYY release) emanating from the hindgut.

Animals↗
PubMed1996

Human monoclonal antibody against colon cancer.

The purpose of the study was to produce human monoclonal antibodies (hMcAb) against human colon cancer for use in radioimmunoimaging. Human-mouse heterohybridomas were developed by fusing SHM-D33 mouse-human hybrid heteromyeloma cells with human lymphocytes from colon cancer tumor-draining lymph nodes. The hybridomas capable of secreting human monoclonal antibodies were screened by using human colon cancer cell lines and pathological biopsies with ELISA and immunohistochemical methods. hMcAb clone H11 was selected for a large-scale antibody production, which was purified from mouse ascites. Biodistribution study demonstrated that specific uptake of 125I-hMcAb H11 by human colon cancer xenografts was significantly higher than by normal tissues. Radioimmunoimaging of human colon cancer xenografts exhibited distinct tumor visualization during the period of 72-96 hr after intraperitoneal injection of 125I-hMcAb H11. The development of human monoclonal antibodies such as hMcAb H11 may be useful for radioimmunodetection and therapy of colon cancer.

Animals↗
PubMed1995

Downregulation of prohormone convertase-1 by a phorbol ester.

Acute TPA treatment (1h, 100nM) of a human pancreatic carcinoid cell line (BON) depletes cell contents of chromogranin A (CGA) and pancreastatin (PST), a peptide derived posttranslationally from CGA. Despite removal of TPA, BON cells continue to release CGA in an unregulated fashion whereas PST secretion is reduced substantially. TPA treatment also reduced prohormone convertase-1 (PC-1) protein and increased PC-1 mRNA levels. Together, these findings indicate that the TPA-induced switch from a regulated to unregulated pattern of CGA secretion is accompanied by a decrease in the processing of CGA to PST and a decrease in the active form of a processing enzyme potentially involved in processing CGA to a smaller peptide, PST.

Aspartic Acid Endopeptidases↗
PubMed1995

Growth of mouse hepatocytes is stimulated by gastrin.

Hepatocyte growth is regulated by various growth factors, including epidermal growth factor (EGF) and insulin. Recently, several additional peptide hormones have been shown to stimulate growth of hepatocyte only in the presence of EGF or insulin and are thus termed secondary mitogens. Gastrin regulates growth of normal and neoplastic gastrointestinal tissues, but the effect on growth of hepatocyte is unknown. We examined the effect of gastrin on growth of a normal mouse hepatocyte (NMH) line established in our laboratory. Effect of gastrin-17 (G-17) (10(-8) to 10(-6) M) on growth of NHM cells was examined in either the presence or absence of EGF in the culture medium. Growth of NMH cells was evaluated by incorporation of either bromodeoxyuridine (BrdU) or 3H-thymidine and by counting cells. Presence of a cell-surface receptor for G-17 was determined by Scatchard analysis using 125I-G-17. In the presence of EGF, gastrin stimulated growth of NMH cells; in the absence of EGF, gastrin did not affect growth. The stimulatory effect of gastrin on NMH cells was blocked by JMV 320, a CCK-B type receptor antagonist. NMH cells possess a single, high affinity binding site for gastrin (Kd = 1.2 nM); EGF increased the gastrin binding capacity compared to non-treated cells (3.5 +/- 0.4 vs. 2.2 +/- 0.6 fmol/10(6) cells). G-17 stimulated growth of NMH cells through a single high affinity receptor for G-17 which pharmcologically appears to be the CCK-B type only in the presence of EGF and thus can be considered a secondary mitogen.

Animals↗
PubMed1995

Secretin potentiates cholecystokinin-stimulated amylase release by AR4-2J cells via a stimulation of phospholipase C.

Alterations in the activity of phospholipase C (PLC) are thought to be the primary intracellular events leading to pancreatic acinar cell exocytosis of zymogen granules. When multiple hormones, each of which may stimulate different signal transduction pathways, bind to cell surface receptors, the cell must integrate these signals into a common response through communication (cross-talk) among intracellular second messengers. We show that cholecystokinin (CCK) induces amylase secretion from AR4-2J pancreatic acinar cells via stimulation of PLC activity. Secretin indirectly stimulated the PLC pathway through cross-talk of the activated cAMP pathway to potentiate the CCK-stimulated amylase secretion. Therefore, secretin potentiated the acinar cell secretory response to CCK by cAMP-mediated cross-talk with the PLC signal transduction pathway.

Amylases↗
PubMed1995

Heat shock proteins are differentially expressed in human gastrointestinal cancers.

The heat shock proteins (Hsp) are stress-responsive genes present in all species; increases of Hsp can confer chemotherapeutic resistance to certain cancers. The purpose of this study was to determine Hsp expression in human gastric, pancreatic and colon cancers. Gastric (n = 3), pancreatic (n = 6) and colon (n = 8) cancers were extracted for RNA and protein, and Northern and Western blots performed. We found that hsp70 and hsp27 mRNA levels were differentially expressed in the gastrointestinal cancers; mRNA expression closely correlated with protein levels suggesting regulation at the level of transcription. In addition, Hsp90 and BiP proteins were constitutively expressed in the gastrointestinal cancers. We conclude that the Hsp are differentially expressed in human gastric, pancreatic and colon cancers; these increases in Hsp occur constitutively and are not the result of physiological or environmental stresses. Increases of Hsp expression in cancer cells may enhance resistance and account for the altered sensitivity of certain gastrointestinal cancers to chemotherapeutic agents.

Gastrointestinal Neoplasms↗
PubMed1995

Nerve growth factor as a mitogen for a pancreatic carcinoid cell line.

Carcinoid tumors are a group of neuroendocrine neoplasms distributed widely throughout the body but most commonly occurring in the gut. These tumors retain many characteristics of their neural crest origin, including secretion of neuroactive peptides and responsiveness to neurotrophic substances. Nerve growth factor (NGF), a neurotrophic protein involved in maintenance and differentiation of peripheral sympathetic and sensory neurons, regulates growth of several neural tumor cells by inducing a differentiated phenotype and subsequent inhibition of cell growth rate. We examined the actions of NGF in a functioning human pancreatic carcinoid cell line (termed BON). NGF has no effect on the cytoarchitecture or constitutive secretion of bioamines in this carcinoid cell line. NGF, however, stimulates the in vitro cellular proliferation of BON cells. BON cells possess mRNA for the NGF receptors (p75LNGFR and p140trkA) and membrane-associated tyrosine kinase activity is increased in response to NGF. Both the mitogenic activity of NGF, as well as the receptor-linked tyrosine kinase activity, can be abrogated in BON cells by the trkA inhibitor K-252a and specific anti-NGF antibody. Our studies demonstrate that NGF is a mitogen for this carcinoid cell line without effect on cellular phenotype or cytoarchitecture. NGF may play a role in the development and progression of human carcinoid tumors.

Carbazoles↗
PubMed1995

Effect of ileo-jejunal transposition on the growth of the GI tract and pancreas in young and aged rats.

BACKGROUND: Ileo-jejunal transposition (IJT; transposition of the distal quarter of the small intestine into the proximal jejunum) is known to stimulate mucosal growth of the transposed ileum, but the effects on other parts of the small intestine are controversial. The effect of aging on the trophic action of IJT is not known. METHODS: We examined the trophic effect of IJT (3 weeks post-operation) on the gastrointestinal tract and pancreas, and on plasma levels of neurotensin and gastrin in three different aged groups of Fischer 344 rats (4, 12, and 24 months old). RESULTS: Three weeks after IJT, the mucosal mass, villus height, and crypt depth increased significantly in the transposed ileum as well as in the remainder of the small intestine. The weights of the colon and pancreas increased significantly after IJT. These responses were not affected by aging. In each of the three age groups, IJT did not affect plasma gastrin level, but significantly increased plasma level of neurotensin. CONCLUSIONS: The distal ileum appears to play an important role in the regulation of growth in the intestine and pancreas; this role is preserved in aged rats. Neurotensin may play an important role in this mechanism.

Aging↗
PubMed1995

Trophic response of gut and pancreas after ileojejunal transposition.

OBJECTIVE: The authors determined whether ileojejunal transposition (IJT) stimulates the growth of the pancreas or the nontransposed segment of small intestine, and ascertained whether this trophic effect is altered by the location of transposed gut segment. SUMMARY BACKGROUND DATA: Transposition of the ileum to the proximal small intestine stimulates a marked mucosal growth of the transposed ileal segment; the cellular mechanisms responsible for this adaptive hyperplasia are not known. METHODS: The distal quarter of the small intestine (distal ileum) was transposed into the proximal (Type I), middle (Type II), or distal (Type III) portions of the remaining small intestine. On postoperative day 28, the pancreas and scraped mucosa from the segments of transposed ileum, proximal ileum, and duodenum were obtained, weighed, and examined for DNA and protein content. RESULTS: All types of IJT increased mucosal weight and DNA content of the transposed ileum. Types I and II IJT produced a significant proliferation of the pancreas and mucosa of the duodenum and proximal ileum. The magnitude of proliferative increases was greatest in Type I IJT. CONCLUSIONS: Ileojejunal transposition appears to be an excellent model to examine the mechanisms by which intestinal epithelial cells proliferate in response to luminal nutrients or humoral factors.

Adaptation, Physiological↗
PubMed1995

Bombesin stimulates mucosal growth in jejunal and ileal Thiry-Vella fistulas.

OBJECTIVE: The authors determined whether the trophic effects of bombesin (BBS) on the small bowel mucosa are mediated by either nonluminal factors or endogenous luminal secretion. SUMMARY BACKGROUND DATA: The gut hormone bombesin stimulates growth of small bowel mucosa. The mechanisms responsible for this trophic effect are not known. METHODS: Rats underwent construction of a Thiry-Vella fistula (TVF) of either the jejunum or ileum. On postoperative day 10, the two groups were subdivided to receive either saline (control) or bombesin (10 micrograms/kg, subcutaneously, three times a day). After 14 days, rats were killed and the TVF was removed. The mucosa was scraped and weighed, and DNA and protein content was determined. RESULTS: Bombesin significantly increased mucosal weight and DNA and protein content of both the jejunal and ileal TVF compared with the control rats. CONCLUSIONS: Bombesin-mediated stimulation of small bowel mucosal growth is mediated by factors that are independent of luminal contents and pancreaticobiliary secretion. Bombesin may prove to be an important enterotrophic factor for gut mucosal proliferation.

Animals↗
PubMed1995

Increases in nup475 and c-jun are early molecular events that precede the adaptive hyperplastic response after small bowel resection.

OBJECTIVE: The authors determined whether increases of nup475 and c-jun gene expression occur after small bowel resection and whether these changes are specific to the gut. SUMMARY BACKGROUND DATA: Massive small bowel resection (SBR) is characterized by adaptive proliferation of the remaining gut mucosa; the molecular signals responsible for this adaptive hyperplasia are unknown. Increases in the "immediate-early genes" nup475 and c-jun are noted in some proliferating tissues; however, alterations in the expression of these genes have not been described in the gut after SBR. METHODS: Rats underwent either a 70% proximal SBR or intestinal transection with reanastomosis (SHAM) and were then killed over a time course (0.5, 2, and 24 hours). The ileum, duodenum, colon, and kidneys were removed and RNA was extracted for Northern hybridization. RESULTS: The authors found that steady-state mRNA levels of both nup475 and c-jun were increased 81% and 62%, respectively, in the ileal remnant at 2 hours in rats after SBR compared with the SHAM group. In addition, nup475 was increased 101% in the duodenum at 24 hours and 31% in the colon at 0.5 hours in rats after SBR. In contrast, neither gene was increased in the kidney. CONCLUSIONS: Increases in steady-state levels of nup475 and c-jun are limited to the gut after SBR, and the timing and magnitude of these changes differ, depending on the gut segment. Finally, the rapid and nutrient-independent increases of nup475 and c-jun suggest an important role for these genes as early molecular signals that participate in the adaptive hyperplasia occurring in the gut remnant after SBR.

Adaptation, Physiological↗
PubMed1995

Role of bombesin on gut mucosal growth.

OBJECTIVE: The authors examined the effects of exogenous bombesin (BBS) on gut mucosal growth in chow-fed rats and the mucosal regeneration after gut atrophy brought about by feeding an elemental diet and after intestinal injury produced by methotrexate (MTX). SUMMARY BACKGROUND DATA: Bombesin is one of many gastrointestinal peptides implicated in the regulation of gut mucosal growth. Although BBS is known to stimulate growth of normal pancreatic tissue, the trophic effect of BBS on gut mucosa is less clear and its exact role in gut mucosal regeneration and repair is not known. METHODS: Rats were fed a regular chow diet (control) or an elemental diet plus either saline or BBS (10 micrograms/kg). In another experiment, rats fed a chow diet and treated with saline or BBS were given MTX (20 micrograms/kg) or a single intraperitoneal injection. In all experiments, small and large bowel mucosa and pancreas were removed and analyzed for BBS-mediated proliferation. RESULTS: Bombesin produced significant mucosal proliferation of the small bowel at day 14, but not at day 7, in rats fed regular chow. In contrast, BBS treatment for 7 days produced significant proliferation in both the atrophic and injured gut mucosa of rats given elemental diet or MTX. CONCLUSIONS: Bombesin may be an important enterotrophic factor for normal mucosal proliferation and may be clinically beneficial as an agent to restore or maintain gut mucosa during periods of atrophy or injury.

Animals↗
PubMed1995

Characterization of promoter elements required for cell-specific expression of the neurotensin/neuromedin N gene in a human endocrine cell line.

Expression of the gene encoding neurotensin/neuromedin N (NT/N) is mostly limited to the brain and specialized enteroendocrine cells (N cells) of the distal small intestine. We have analyzed the NT/N DNA sequences upstream of the RNA start site that direct cell-specific expression using a novel human endocrine cell line, BON, that resembles intestinal N cells in several important aspects, including NT/N precursor protein processing, ratios of different NT/N mRNA forms, and high levels of constitutive expression of the NT/N gene. Transient transfection assays with plasmids with progressive 5' deletions of the rat NT/N promoter identified the proximal 216 bp of 5' flanking sequences as essential for high-level constitutive NT/N expression in BON cells. In addition, a detailed mutational analysis defined multiple regions within the proximal 216 bp that contribute to cell-specific NT/N expression. These elements include a proximal cyclic AMP response element (CRE)/AP-1-like motif (TGACATCA) that binds c-Jun, JunD, CRE-binding (CREB), and ATF proteins, a near-consensus glucocorticoid response element, and a distal consensus AP-1 site that binds c-Fos, Fra-1, and JunD. In addition, elements contained within two 21-bp imperfect direct repeats play an important role in NT/N expression in BON cells and may bind novel factors that act as positive regulators of NT/N expression. DNase I footprinting and gel shift analyses demonstrate that the sites identified by mutational analysis, and at least one additional site, specifically bind BON cell nuclear proteins in vitro. We speculate that a complex pattern of regulation requiring interaction between a proximal CRE/AP-1-like motif and other upstream control elements play an important role in the high-level constitutive expression of NT/N in the human endocrine cell line BON. In addition, the BON cell line provides a unique model to further characterize the factors regulating cell-specific NT/N expression and to better understand the mechanisms responsible for the terminal differentiation of the N-cell lineage in the gut.

Activating Transcription Factor 1↗
PubMed1995

Differential regulation by TGF-beta 1 and insulin of insulin-like growth factor binding protein-2 in IEC-6 cells.

The purposes of this study were to determine the regulation of insulin-like growth factor binding protein-2 (IGFBP-2) in IEC-6 cells by transforming growth factor-beta 1 (TGF-beta 1) and insulin and to determine whether IGFBP-2 mediated the growth-inhibitory action on the cells. Utilizing Western ligand blot analysis, we found that TGF-beta 1 at concentrations of 0.5, 1.0, and 2 ng/ml significantly increased levels of 32-kDa IGFBP in the conditioned medium (CM) of IEC-6 cells in a dose-dependent fashion and that low doses of insulin (1.0 and 5.0 microgram/ml) also increased IGFBP levels in the CM of IEC-6 cells, but a high dose of insulin (10 micrograms/ml) depressed IGFBP release in the CM. Immunoblotting has shown that the IGFBP of 32 kDa was IGFBP-2 and further confirmed the above results. IGFBP-2 mRNA levels were stimulated by TGF-beta 1 (2.0 ng/ml) and suppressed by insulin (5.0 micrograms/ml). In addition, des (1-3) IGF-I (50 ng/ml) and insulin stimulated the proliferation of IEC-6 cells. Anti-IGFBP-2 antibodies partially blocked the inhibitory role in IEC-6 cell growth evoked by des (1-3) IGF-I. These findings suggest that the upregulation of IGFBP-2 by TGF-beta 1 occurs, at least in part, at the level of mRNA, whereas the regulation by insulin appears to be at a posttranslational level, and that the TGF-beta 1-stimulated production of IGFBP may contribute to the growth-inhibitory action in intestinal epithelial cells.

Animals↗
PubMed1995

Intestinal peptide YY: ontogeny of gene expression in rat bowel and trophic actions on rat and mouse bowel.

The purpose of this study was twofold: 1) to characterize the profile of colonic peptide YY (PYY) gene expression in rats and 2) to examine for potential trophic effects of PYY on the intestine in rats and mice. Expression of PYY mRNA (analyzed by Northern blotting and in situ hybridization) and PYY (analyzed by high-performance liquid chromatography and radioimmunoassay) was detected initially at day 17 of gestation in colonic extracts of Sprague-Dawley and Fischer rats. Expression of colonic PYY mRNA increased until 7 days of age and remained at its highest level (approximately twofold greater than the adult level) through the end of the nursing period. After weaning (21 days of age), PYY mRNA levels declined quickly to adult levels. Colonic PYY concentrations followed, in a coordinated manner, with some temporal delay after birth, the increase and decrease of its mRNA. Administration of PYY increased the weight and DNA content of the duodenum significantly in nursing rats and adult mice. In mice, PYY treatment also increased weight and DNA content of the ileum and colon. The trophic effects of PYY were dose related, peptide specific, and independent of species and sex. From these findings, we hypothesize that PYY plays an important role in intestinal development and dietary adaptation.

Aging↗
PubMed1995

Phorbol ester-induced alteration in the pattern of secretion and storage of chromogranin A and neurotensin in a human pancreatic carcinoid cell line.

Brief phorbol ester treatment of BON cells results in a persistent release and cellular depletion of immunoreactive chromogranin A (CGA-IR) and neurotensin (NT-IR) cell contents. The purpose of the present study was to characterize the effects of 12-O-tetradecanoyl phorbol-13-acetate (TPA) on the secretion, biosynthesis, and steady-state messenger RNA (mRNA) levels of chromogranin A (CGA) and of a coresident peptide, neurotensin, by a novel human pancreatic carcinoid cell line, called BON. Acute TPA treatment (100 nM, 1 h) of BON cells resulted in 20- and 40-fold elevations in release of CGA-IR and NT-IR, respectively; and a 70-90% depletion of CGA-IR and NT-IR cell contents. TPA treatment also increased the biosynthetic rate of CGA-IR. Steady-state mRNA levels of CGA and NT/N (neurotensin/neuromedin N) were unchanged. Cell contents of CGA-IR and NT-IR were not replenished for a period of up to 6 days; secretion of CGA-IR and NT-IR persisted. In addition, BON cells failed to release CGA in response to stimulation by ionomycin and A23187 several days after acute TPA treatment. Our data indicate that the lack of replenishment of cell contents of CGA-IR and NT-IR is not due to decreases in steady-state CGA-IR and NT-IR mRNA levels, nor is it due to a decrease in biosynthesis of CGA-IR, but it is the result of a loss in the ability of TPA-treated BON cells to store and secrete CGA-IR and NT-IR in a regulated manner. These effects of TPA are mediated through the PKC pathway.

Analysis of Variance↗