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Ghrelin, a nature ligand for the growth hormone secretagogue receptor (GHS-R), stimulates a release of growth hormone, prolactin and adrenocorticotropic hormone. Also, ghrelin increases food intake in adult rats and humans and exhibits gastroprotective effect against experimental ulcers induced by ethanol or stress. The aim of present study was to examine the influence of ghrelin administration on gastric and duodenal growth and expression of pepsin and enterokinase in young mature rats with intact or removed pituitary. Methods: Two week after sham operation or hypophysectomy, eight week old Wistar male rats were treated with saline (control) or ghrelin (4, 8 or 16 nmol/kg/dose) i.p. twice a day for 4 days. Expression of pepsin in the stomach and enterokinase in the duodenum was evaluated by real-time PCR. Results: In animals with intact pituitary, treatment with ghrelin increased food intake, body weight gain and serum level of growth hormone and insulin-like growth factor-1 (IGF-1). These effects were accompanied with stimulation of gastric and duodenal growth. It was recognized as the significant increase in gastric and duodenal weight and mucosal DNA synthesis. In both organs, ghrelin administered at the dose of 8 nmol/kg caused maximal growth-promoting effect. In contrast to these growth-promoting effects, administration of ghrelin reduced expression of mRNA for pepsin in the stomach and was without effect on expression of mRNA for enterokinase in the duodenum. Hypophysectomy alone lowered serum concentration of growth hormone under the detection limit and reduced serum level of IGF-1 by 90%. These effects were associated with reduction in daily food intake, body weight gain and gastroduodenal growth. In hypophysectomized rats, administration of ghrelin was without significant effect on food intake, body weight gain or growth of gastroduodenal mucosa. Also, serum concentration of growth hormone or IGF-1 was not affected by ghrelin administration in rats with removed pituitary. Conclusion: Administration of ghrelin stimulates gastric and duodenal growth in young mature rats with intact pituitary, but inhibits expression of mRNA for pepsin in the stomach. Growth hormone and insulin-like growth factor-1 play an essential role in growth-promoting effects of ghrelin in the stomach and duodenum.
Background. Yeast extract powder (YEP) is a rich source of several B vitamins and is used mainly in bacterial culture. In the present study, the effect of dietary supplementation of YEP on growth, feed conversion, nutrient ADC, body composition, and digestive enzyme profile of rohu, Labeo rohita (Hamilton, 1822) fingerlings was evaluated. Materials and Methods. Rohu fingerlings (mean individual weight 1.76 ± 0.15 g) were fed, twice daily, fish meal-based 35% protein diets supplemented with YEP (0.1%, 0.2%, 0.3%, 0.4%, and 0.5% on dry matter basis) for 75 days at 3% of body weight, in triplicate. Fish performance in terms of growth, apparent nutrient digestibility, and whole body composition was studied. The intestinal protease and amylase activities were also determined in rohu fingerlings fed YEP-supplemented diets. Microbial cultures of the intestinal mucosa of the fish in all dietary treatments were carried out after the feeding trial in soybean-casein digest agar (TSA), gelatin-peptone (GP), starch (ST) and carboxymethylcellulose (CMC) plates, separately, to determine the protease- amylase-, and cellulase-producing capacities (qualitative) of the isolated organisms. Results. The fish fed YEP-supplemented diets exhibited better growth than the control up to 0.2% level. YEP at 0.1% level resulted in the best performance of the fish in terms of per cent weight gain, SGR, FCR, and PER, followed by the 0.2% level. YEP led to higher deposition of protein and lower deposition of lipid in the carcass. The intestinal protease activity was highest in the fish fed 0.1% yeast, followed by 0.2%. Protease-producing capacity of the intestinal bacterial flora in the in vitro cultures followed the same trend. Conclusion. It appears therefore that YEP in the diets might have some effect on the gut microflora by supplying additional digestive enzymes leading to better nutrient utilization.
Rab3D is a low molecular weight GTP-binding protein that associates with secretory granules in exocrine cells. AR42J cells are derived from rat pancreatic exocrine tumor cells and develop an acinar cell-like phenotype when treated with dexamethasone (Dex). In the present study, we examined the role of Rab3D in Dex-treated AR42J cells. Rab3D expression and localization were analyzed by subcellular fractionation and immunoblotting. The role of Rab3D was examined by overexpressing myc-labeled wild-type-Rab3D and a constitutively active form of Rab3D (Rab3D-Q81L) in AR42J cells. We found that Rab3D is predominantly membrane-associated in AR42J cells and co-localizes with zymogen granules (ZG). Following CCK-8-induced exocytosis, amylase-positive ZGs appeared to move towards the periphery of the cell and co-localization between Rab3D and amylase was less complete when compared to basal conditions. Overexpression of WT, but not mutant Rab3D, resulted in an increase in cellular amylase levels. Overexpression of mutant and WT Rab3D did not affect granule morphology, CCK-8-induced secretion, long-term (48 hr) basal amylase release or granule density. We conclude that Rab3D is not involved in agonist-induced exocytosis in AR42J cells. Instead, Rab3D may regulate amylase content in these cells.
Probiotic strains of L. casei, L. acidophilus, L. helveticus, B. animalis, B. lactis and two strains of B. bifidum were examined for their survival at pH 3.0 and 3% bile salts. Additionally, the viability of L. casei cells was tested in presence of pepsin, trypsin, chymotrypsin, alpha-amylase and lipase, and after heating at 90ºC for 30 min. It was found that all pre-treatments caused significant decrease in bacteria survival. The most sensitive strain for stress factors was B. bifidum 1. Using Caco-2 monolayer culture as the in vitro adhesion model, the significant reduction of adherence after thermal treatment, digestion with pepsin, trypsin and chymotrypsin, and bile salts were observed. The inhibition of bacteria adhesion after thermal denaturation and proteolysis proved an hypothesis that adhesion factors are of proteinaceous origin.
Protein sparing by lipid has been demonstrated in certain cultivable species of fish. This study was carried out using four low protein isonitrogenous diets (24% crude protein) formulated by supplementing varied levels of fish oil (0, 3, 6, and 9%). The diets were fed for 120 days at 5% body weight to triplicate groups of common carp (av. wt. 2.13-2.21 g) stocked at 1 per m2 in mud bottomed cement tanks (18 m2), fertilized with poultry manure. The growth of fish was the highest (P < 0.05) with the diet containing 6% fish oil, followed by 3, 9, and 0%. Food conversion ratio and protein efficiency ratio improved with increasing dietary lipid level. Dietary lipid had a positive impact on carcass lipid level (P < 0.05). Moisture and crude protein did not vary (P > 0.05) from that of the control. Survival ranged from 96.29 (T1) to 100% (T0) without any significant (P > 0.05) difference among the treatments. While there was a general increase in amylase activity in the treated fish, protease activity showed a reduction with increase in oil supplementation. No difference (P > 0.05) in lipase activity was observed between the different treatments. The results indicate the beneficial effects of incorporating fish oil in the diet of common carp.
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