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Renewal of parietal cells, chief cells, and smooth muscle cells in mouse stomach
April 1998 contraction ( -+ SEM) when integrated over time was low in response to both sucrose polyester (-150 +-214 mL * 120 min) and saline (-89 -+ 123 mL * 120min). In contrast, there was a marked contraction in response to digestible fat (1069--.253 mL * 120 min). Sucrose polyester did not affect plasma cholecystokinin concentrations (-9.3- 15.0 pmol/L * 120 min) whereas digestible fat resulted in a significant increase (89.5 -+44.8 pmol/L * 120 min, p=0.014) compared to saline (-3.0 +- 13.8 pmol/L * 120min). Conclusion: Ingestion of sucrose polyester, in contrast to digestible fat, falls to stimulate gallbladder contraction or to stimulate release of cholecystokinin. G3661 TRANSFORMING GROWTH FACTOR ALPHA KNOCKOUT MICE HAVE SMALLER SMALL INTESTINES, LARGER LARGE INTESTINES BUT NO INCREASED SENSITIVITY TO NSAIDINDUCED SMALL INTESTINAL INJURY. CE Macdonald. RJ Playford*, M Khatri*, RA Goodlad**. Department and University Division of Gastroenterology*, Leicester General Hospital NHS Trust, Gwendolen Rd., Leicester and Imperial Cancer Research Fund, 44 Lincon's Inn Fields, Holbum, London.**, UK. Introduction: Transforming Growth Factor Alpha (TGF~) is a mitogenic peptide produced in the mucosa throughout the gastrointestinal tract. TGF~ knockout mice have been reported to have a GI tract which appears phenotypically normal (Cell 1993 73,249-261). A recent study, however, suggests their colons have increased sensitivity to DSS induced colitis (Gastroenterology 1997 113,825-832). We therefore performed detailed analysis of gut morphology and morphometry under basal circumstances and examined the sensitivity of the small intestine to injury induced by the NSAID, indomethacin. Methods: TGF~ knockout, heterozygous and wild type mice (all CBA strain) (n=14) were all born within five days of each other and assessed at 12 weeks of age. Animals received indomethacin 85mg/kg so. or saline 24 hours before killing. In addition, proliferation was assessed in all animals using vincristine induced metaphase arrest (lmg/kg ip.). Wet weights of internal organs and length of small and large intestine were assessed followed by microdissection, metaphase counting and villus height assessment. Analyses used ANOVA. Results: Across all groups males were heavier with heavier intestinal tracts. Knockout mice displayed phenotypic abnormalities of wavy hair and corneal inflammation previously described. Compared to controls, the total body weight of knockout mice was 7% lighter with a 10-20% reduction in stomach weight, small intestinal length and small intestinal weight when expressed as absolute weight or as percentage of total body weight (all p<0.01 vs. control). In contrast, knockout mice had about 10% increase in colonic length and weight. (p<0.01 vs. control). Heterozygotes showed intermediate results in all the parameters, lndomethacin caused a 14% reduction in villus heights across all the groups. Conclusions: TGFcx plays a role in the growth of the gastrointestinal tract. TGFct does not increase the sensitivity of the small intestine to NSAID induced damage, other factors may therefore be more important in maintaining small intestinal integrity. G3662 RENEWAL OF PARIETAL CELLS, CHIEF CELLS, AND SMOOTH MUSCLE CELLS IN MOUSE STOMACH. Y. Magami, S. Kokuno, M. Furukawa, Y. Tsukioka, D. Nakayama, T. Azuma, H. Inokuchi, K. Kawai, T. Hattori, T. Saitoh, Dept. of Internal Medicine, Tokyo Medical College Hospital, Tokyo, Dept. of Preventive Medicine, Kyoto Prefectural University of Medicine, Kyoto, Dept. of Pathology, Shiga University of Medical Science, Shiga, Japan.
Since [3H]thymidine autoradiography was first applied to the study of cell kinetics, the epithelial cells of gastrointestinal mucosa have been extensively examined. Renewal rate of most types of glandular cells in the gastric mucosa have been estimated. However, renewal rate of parietal cells and chief cells bad not been estimated precisely, because that of their cells are very slow. In the past we reported on cell renewal of these cells in mouse gastric mucosa in this meeting. The half population of parietal cells and chief cells renew for about 100 days and 200 days, respectively. It was reported that smooth muscle cells of the alimentary tract was classified as "expanding cell population". Renewal rate of smooth muscle cells have not yet been estimated precisely. The present study was designed to analyze the renewal of smooth muscle cells by [3H]thymidine autoradiography. In order to estimate the turnover rate, we used the pulse labeling method after the long continuous labeling with [3H]thymidine. The animals received 112 repeated injections of [3H]thymidine at 6hr. intervals for 28 days after birth and were killed on lst., 60th, 120th, 200th, 300th day after the last injection. Immediately after killing the animals, the fundic portion of the stomach was fixed and embedded in paraffin. The sections were dipped in nuclear emulsion and developed after 5 weeks exposure and finally stained with hematoxylin and eosin. After 112 repeated continuous labeling, most of smooth muscle cells in stomach were labeled. This in dicates that most of smooth muscle cells arise in postnatal day. We scored grain counts on nuclei of smooth muscle ceils. By the pulse labeling, it was found that some of labeled cells were replaced by unlabeled cells and weakly labeled cells, but many heavily labeled cells remained. Mean grain counts decreased to 70% for 300 day. This finding indicates that smooth muscle cells renew very slowly. Ithe period in which mean grain counts decrease to 50%, is estimated as 600 days. The renewal rate of smooth muscle cells is much more slow than that of parietal cells or chief cells.
Growth, Development, Nutrition A893
• G3663 LONGITUDINAL CHANGES IN LIVER ENZYMES AND GROWTH STATUS IN CHILDREN WITH CYSTIC FIBROSIS. MR Mascarenhas BS Zemel, ES Mallet, TF Scanlin, VA Stallings, Department of Pediatrics, Univ. of. Pennsylvania, Philadelphia, PA. The incidence of hepatobiliary disease in CF is variable and increases with age, but few studies have examined longitudinal changes in liver function tests (LFTs) and their association with other measures of health outcome, such as growth. We evaluated LFTs (ALl', AST, GGT, bilirubin) and growth in a group of 24 children (5-10 yrs old) enrolled in a 4 yr longitudinal study. The sample was subsequently followed for several yrs to determine long term patterns of growth and liver function. The average total number of annual measurements was 7.6 (range 3-9). LFTs were defined as elevated if they were above the normal range for age and gender and abnormal if they were twice the upper limit of the normal range. Persistence of elevated or abnormal values was defined as _>50% of observations for that individual. Height (Ht) and Weight (Wt) were measured and Z-scores were generated from National Center for Health Statistics reference values. Nineteen children had elevated ALT, including 10 with persistently elevated ALT levels. Six children had abnormal ALT, including 2 with persistently abnormal ALT. Nineteen had elevated AST levels, including 7 with persistently elevated levels, but none were abnormal. Eight children had elevated GGT, including 5 with persistently elevated GGT. Four children had abnormal GGT, including 2 with persistently abnormal levels. Two had elevated bilimbin, 1 with persistently elevated levels, but neither were in the abnormal range. Two of the children with persistently abnormal GGT developed cirrhosis with portal hypertension and splenomegaly. For the entire group, Wt and Ht Z-scores declined significantly (p<0.0005) over time (Ht-Z::-005 + 0.01; Wt-Z: -0.06-+ 0.01 units per yr). On average, the group with persistently elevated GGT levels had poorer growth status (Ht-Z: -1.0-+ 1.1 vs -0.3 ± 0.7; Wt-Z: -1.0 -+0.9 vs -0.3 + 0.7), but these differences were not statistically significant. These findings show frequently elevated ALl', AST and GGT levels in some children with CF. GGT may be the best longitudinal predictor of chronic liver disease. Growth status declined for the group overall, but longitudinally, there were no associations between LFT results and growth status. Research was supported by the CF Foundation, the GCRC (M01RR00240) and the Nutrition Center of the Children's Hospital of Philadelphia. G3664 NEONATAL RENAL INSUFFICIENCY AND RENAL ACIDOSIS ARE MANIFESTATIONS OF ALAGILLE SYNDROME IN INFANCY. KE McBride, DA Piceoli. The Children's Hospital of Philadelphia, University of Pennsylvania, Philadelphia, PA.
AGS has been defined as bile duct paucity associated with the syndromic features of cholestasis, heart murmur, vertebral anomalies, posterior embryotoxon and peculiar facies. Renal disease, both structural and functional, including adult onset renal failure has been reported. Rarely, progressive neonatal renal failure has lead to early mortality. Other infants with AGS have glomerular lesions (mesangiolipidosis) without clinical renal disease. Neonatal renal insufficiency with resolving renal acidosis is not described. Methods: The records of 92 patients with AGS were reviewed. Results: Of these, 5 neonates (4M, 1F) had evidence of renal insufficiency. Mean birth weight 2.2 kg was at a mean gestational age of 35.7 weeks, and 3 of 5 had intrauterine growth retardation (IUGR). All five infants had elevated creatinine (mean 1.8 mg/dl, range 1.4 to 2.7). In four this was identified in the first week of life. Metabolic acidosis was present in each infant, with low serum bicarbonate levels (mean 13 mg/dl, range 8 to 17). In-utero ultrasounds in two infants with IUGR demonstrated "polycystic kidney disease" in one and hydronephrosis with oligohydramnios in one. A third infant with in-utero oligohydramnios was IUGR. Proteinuria was observed in two patients on urinalysis. In all patients the diagnosis of AGS was made subsequent to the renal presentation (mean age of diagnosis 16 months, range 2 months to 6 years old). Four patients had significant difficulty with poor weight gain. All patients responded to supplimental sodium bicarbonate or citrate with normalization of the acidosis and improved weight gain. 3 patients were weaned from the base supplimentation by 2 months old. The average creatinine at I week of age was 1.7 (1.6 to 2.7) and by 4 to 6 months was 0.5 (0.2 to 0.9). All patients had improved renal function. Renal ultrasonography demonstrated bilateral small and echogenic kidneys in three cases, bilateral UPJ in one case and one atrophic kidney in another case. The renal biopsy of the female patient revealed findings consistent with nephronophthisis. Autopsy findings on the patient who died at 2 years old of unclear causes (but not renal failure) revealed multiple cystic spaces lined by euboidal epithelium. Glomerulolipidosis was noted and a cartilagenous mass (hamartoma) is also noted in the kidney. Conclusions: In addition to the renal disease in children and older patients, renal insufficiency and renal acidosis occur in neonates with AGS. These manifestations may precede the diagnosis of AGS, and contribute to IUGR and poor post-natal growth. Therapy is effective, and functional abnormalities may be transient. The variety of structural and functional abnormalities in AGS renal disease is as diverse as the range of cardiac malformations. Renal disease should be considered as a syndromic feature of AGS.
Since [3H]thymidine autoradiography was first applied to the study of cell kinetics, the epithelial cells of gastrointestinal mucosa have been extensively examined. Renewal rate of most types of glandular cells in the gastric mucosa have been estimated. However, renewal rate of parietal cells and chief cells bad not been estimated precisely, because that of their cells are very slow. In the past we reported on cell renewal of these cells in mouse gastric mucosa in this meeting. The half population of parietal cells and chief cells renew for about 100 days and 200 days, respectively. It was reported that smooth muscle cells of the alimentary tract was classified as "expanding cell population". Renewal rate of smooth muscle cells have not yet been estimated precisely. The present study was designed to analyze the renewal of smooth muscle cells by [3H]thymidine autoradiography. In order to estimate the turnover rate, we used the pulse labeling method after the long continuous labeling with [3H]thymidine. The animals received 112 repeated injections of [3H]thymidine at 6hr. intervals for 28 days after birth and were killed on lst., 60th, 120th, 200th, 300th day after the last injection. Immediately after killing the animals, the fundic portion of the stomach was fixed and embedded in paraffin. The sections were dipped in nuclear emulsion and developed after 5 weeks exposure and finally stained with hematoxylin and eosin. After 112 repeated continuous labeling, most of smooth muscle cells in stomach were labeled. This in dicates that most of smooth muscle cells arise in postnatal day. We scored grain counts on nuclei of smooth muscle ceils. By the pulse labeling, it was found that some of labeled cells were replaced by unlabeled cells and weakly labeled cells, but many heavily labeled cells remained. Mean grain counts decreased to 70% for 300 day. This finding indicates that smooth muscle cells renew very slowly. Ithe period in which mean grain counts decrease to 50%, is estimated as 600 days. The renewal rate of smooth muscle cells is much more slow than that of parietal cells or chief cells.
Growth, Development, Nutrition A893
• G3663 LONGITUDINAL CHANGES IN LIVER ENZYMES AND GROWTH STATUS IN CHILDREN WITH CYSTIC FIBROSIS. MR Mascarenhas BS Zemel, ES Mallet, TF Scanlin, VA Stallings, Department of Pediatrics, Univ. of. Pennsylvania, Philadelphia, PA. The incidence of hepatobiliary disease in CF is variable and increases with age, but few studies have examined longitudinal changes in liver function tests (LFTs) and their association with other measures of health outcome, such as growth. We evaluated LFTs (ALl', AST, GGT, bilirubin) and growth in a group of 24 children (5-10 yrs old) enrolled in a 4 yr longitudinal study. The sample was subsequently followed for several yrs to determine long term patterns of growth and liver function. The average total number of annual measurements was 7.6 (range 3-9). LFTs were defined as elevated if they were above the normal range for age and gender and abnormal if they were twice the upper limit of the normal range. Persistence of elevated or abnormal values was defined as _>50% of observations for that individual. Height (Ht) and Weight (Wt) were measured and Z-scores were generated from National Center for Health Statistics reference values. Nineteen children had elevated ALT, including 10 with persistently elevated ALT levels. Six children had abnormal ALT, including 2 with persistently abnormal ALT. Nineteen had elevated AST levels, including 7 with persistently elevated levels, but none were abnormal. Eight children had elevated GGT, including 5 with persistently elevated GGT. Four children had abnormal GGT, including 2 with persistently abnormal levels. Two had elevated bilimbin, 1 with persistently elevated levels, but neither were in the abnormal range. Two of the children with persistently abnormal GGT developed cirrhosis with portal hypertension and splenomegaly. For the entire group, Wt and Ht Z-scores declined significantly (p<0.0005) over time (Ht-Z::-005 + 0.01; Wt-Z: -0.06-+ 0.01 units per yr). On average, the group with persistently elevated GGT levels had poorer growth status (Ht-Z: -1.0-+ 1.1 vs -0.3 ± 0.7; Wt-Z: -1.0 -+0.9 vs -0.3 + 0.7), but these differences were not statistically significant. These findings show frequently elevated ALl', AST and GGT levels in some children with CF. GGT may be the best longitudinal predictor of chronic liver disease. Growth status declined for the group overall, but longitudinally, there were no associations between LFT results and growth status. Research was supported by the CF Foundation, the GCRC (M01RR00240) and the Nutrition Center of the Children's Hospital of Philadelphia. G3664 NEONATAL RENAL INSUFFICIENCY AND RENAL ACIDOSIS ARE MANIFESTATIONS OF ALAGILLE SYNDROME IN INFANCY. KE McBride, DA Piceoli. The Children's Hospital of Philadelphia, University of Pennsylvania, Philadelphia, PA.
AGS has been defined as bile duct paucity associated with the syndromic features of cholestasis, heart murmur, vertebral anomalies, posterior embryotoxon and peculiar facies. Renal disease, both structural and functional, including adult onset renal failure has been reported. Rarely, progressive neonatal renal failure has lead to early mortality. Other infants with AGS have glomerular lesions (mesangiolipidosis) without clinical renal disease. Neonatal renal insufficiency with resolving renal acidosis is not described. Methods: The records of 92 patients with AGS were reviewed. Results: Of these, 5 neonates (4M, 1F) had evidence of renal insufficiency. Mean birth weight 2.2 kg was at a mean gestational age of 35.7 weeks, and 3 of 5 had intrauterine growth retardation (IUGR). All five infants had elevated creatinine (mean 1.8 mg/dl, range 1.4 to 2.7). In four this was identified in the first week of life. Metabolic acidosis was present in each infant, with low serum bicarbonate levels (mean 13 mg/dl, range 8 to 17). In-utero ultrasounds in two infants with IUGR demonstrated "polycystic kidney disease" in one and hydronephrosis with oligohydramnios in one. A third infant with in-utero oligohydramnios was IUGR. Proteinuria was observed in two patients on urinalysis. In all patients the diagnosis of AGS was made subsequent to the renal presentation (mean age of diagnosis 16 months, range 2 months to 6 years old). Four patients had significant difficulty with poor weight gain. All patients responded to supplimental sodium bicarbonate or citrate with normalization of the acidosis and improved weight gain. 3 patients were weaned from the base supplimentation by 2 months old. The average creatinine at I week of age was 1.7 (1.6 to 2.7) and by 4 to 6 months was 0.5 (0.2 to 0.9). All patients had improved renal function. Renal ultrasonography demonstrated bilateral small and echogenic kidneys in three cases, bilateral UPJ in one case and one atrophic kidney in another case. The renal biopsy of the female patient revealed findings consistent with nephronophthisis. Autopsy findings on the patient who died at 2 years old of unclear causes (but not renal failure) revealed multiple cystic spaces lined by euboidal epithelium. Glomerulolipidosis was noted and a cartilagenous mass (hamartoma) is also noted in the kidney. Conclusions: In addition to the renal disease in children and older patients, renal insufficiency and renal acidosis occur in neonates with AGS. These manifestations may precede the diagnosis of AGS, and contribute to IUGR and poor post-natal growth. Therapy is effective, and functional abnormalities may be transient. The variety of structural and functional abnormalities in AGS renal disease is as diverse as the range of cardiac malformations. Renal disease should be considered as a syndromic feature of AGS.