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Into the unknown - the death pathways in the neonatal gut epithelium

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Apoptosis is a fundamental process in the development of the fast growing intestinal mucosa. Apoptotic cells are present along the whole length of the villi and in the crypts. The mechanisms involved in the induction of apoptosis in the gut mucosa are still unknown. Cytokines are believed to play a role in auto- and paracrine models because the cells are dying in so-called "packets" containing neighboring cells. In the rapidly developing gut of neonates, the apoptosis rate is transiently reduced in the first days of life, enhancing the growth of mucosa. Afterwards, apoptosis plays a role in the exchange of the enterocyte population, facilitating maturation of the mucosa. The presence of autophagic cells has been confirmed for the first time in the developing gut. Deprivation of growth factors during feeding artificial milk formula led to an increased apoptosis rate. Supplementation with leptin reduced cell apoptosis and increased the mitosis-to-apoptosis ratio. Autophagy was also diminished. The key to healthy gut mucosa growth in early life, especially in fast-growing animals, is colostrum, which supplies nutritional and defensive components together with supplementary growth factors, cytokines and hormones essential for growth and maturation of gut mucosa.
Studies were carried out to elucidate the anti-oxidative effect(s) of putrescine (10 mg/kg b.w./day) in rats treated per os with either sodium nitrite (10 mg/kg b.w./day) or normal saline (control) for 14 days. The putrescine was given to rats for 7 days only (days 7-14) and it was introduced 3-4 hrs after nitrite or saline dosage. Sodium nitrite increased thiobarbituric acid reactive substances (TBARS) in rat small intestinal mucosa and liver, and the agent did not have any effect on the total anti-oxidant status (TAS) and lipid peroxidation of rat blood. Nitrite did not also change the activity of superoxide dismutase (SOD) in the small intestinal mucosa, liver and blood, as well. Pretreatment of nitrite-treated rats with putrescine decreased TBARS and increased TAS in animals. Putrescine decreased SOD activity in the blood and liver of nitrite- and/or saline-treated rats, however, the agent did not affect the SOD enzyme in the small intestinal mucosa. Results suggest that putrescine dosed to nitrite-treated rats possesses some anti-oxidative properties.
The experiment was performed on piglets, divided into control and experimental groups The experimental group received orally, from the birth to the 35th d of life, 0.4 g/kg b.w. /d of L-alanyl-L-glutamine dipeptide (Ala-Gln). One week after weaning the piglets were killed and the small intestine and bones were sampled for histological analyses. Measurements of bone physical and geometric properties were performed according to Ferreti method. The mineral density was analysed by the DEXA method. Ala-Gin treated piglets had a higher body weight at the 35 d of age compared to that of the control piglets. Mucosa thickness, villus length, and crypt depth in the jejunum of the piglets showed higher values compared to controls. In Ala-Gin treated piglets bone physical and geometric parameters and mineral density were significantly higher, and the bone structure revealed a shift in its organisation and mineralization process. In conclusion, oral administration of Ala-Gln protects the piglets from body weight loss and intestinal hypotrophy correlated with weaning and preserved the normal development of the femora during the post-weaning period.
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Melatonin-induced protein synthesis in the rat parotid gland

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Melatonin occurs in large amounts in the intestinal mucosa and is released during a meal. Recent studies of ours reveal that exogenous melatonin evokes the in vivo secretion of protein and amylase from the rat parotid gland. The aim of the present study was to investigate the effect of melatonin on the protein synthesis of the parotid gland of pentobarbitone-anaesthetised rats as estimated by the rate of incorporation of [3H]leucine into trichloroacetic acid-insoluble material of the gland. Compared with the parotid protein synthesis (set at 100%) of those rats exposed to an intravenous infusion of melatonin (25 mg/kg during 1 hour), under muscarinic and - and ß-adrenoceptor blockade, the synthesis in the corresponding glands of saline-treated control rats was less (by 25%). The synthesis was also less when the melatonin administration was combined with the melatonin 2-preferring receptor antagonist luzindole (24%), the non-selective nitric oxide synthase inhibitor L-NAME (18%) and the neuronal nitric oxide synthase inhibitor N-PLA (21%). Almost all the melatonin receptor-mediated effect was due to nitric oxide generation via the activity of neuronal type nitric oxide synthase. The present findings lend further weight to the idea that salivary glandular activity associated with food intake is hormonally influenced and they also suggest clinical implications for melatonin in the treatment of xerostomia. Since melatonin is known to exert anti-inflammatory actions in the oral cavity, the stimulatory effect of melatonin may include the synthesis of proteins of importance for the oral defence.
The use of soybean in human and animal nutrition is limited because of high content of bioactive compounds: enzyme inhibitors, polyphenols, goitrogens, phytates, saponins, sugars, and agglutinins. The damage of intestinal mucosa structure was previously observed in animals fed soybean supplemented diets. Hence, the objectives of the presented study were to compare intensity of epithelium remodeling processes in different intestinal segments, and to evaluate the influence of the 1% of soybean dietary supplementation on the processes in intestinal mucosa. The experiment was performed on 30 Wistar rats fed AIN-93 based diets. Animals were divided randomly into three groups: control (CTRL), with 1% of raw soybean (RS) and with 1% of soaked and boiled soybean (BS). The samples of: duodenum (DUO), proximal jejunum (PROX), mid-jejunum (MID), distal-jejunum (DIST) and ileum (ILE) were collected. The following processes in these samples were evaluated: mitosis (Ki-67), apoptosis (Cpp32), autophagy (MAP I LC3) and DNA damage (p53). Present data show that modification of soybean by soaking and subsequent boiling markedly influences the enterocyte turnover in the small intestine mucosa. Increased mitotic ratio in the intestine of rats fed with boiled soybean masks the negative effects of soybean on the small intestine structure.
An interdisciplinary research group granted by the German Research Foundation (FOR 438) tested various hypotheses and tired to develop a model for the mode of action of probiotics in pigs. The study included the fields of animal nutrition/digestion physiology, anatomy and histology of the intestinal mucosa, transport and secretory properties of the mucosa, microbiology of the intestinal tract, immune system (classes of intraepithelial lymphocytes, humoral responses), gene expression of the mucosa and finally the in vitro and in vivo resistance against infection with Salmonella. Five trials with ten sows per treatment each and their piglets and two probiotic strains were included in this study. The studied bacterial strains were Enterococcus faecium NCIMB 10415 and Bacillus cereus var. toyoi NCIMB 40112. Concluding from our studies and the published data of others, the effects of probiotics on performance are rarely significant. However, with one exception the incidence of post-weaning diarrhoea under the effect of both probiotics was significantly reduced in the trials of the research group. Furthermore, the identification frequency of various E. coli sero-pathovars relevant in post weaning diarrhoea was reduced in these animals. On the other hand, no significant modifications were found for the morphology and histology of the intestinal mucosa and also not on transport properties of this tissue. A further important finding was that the mode of action for probiotics is not unique but species or even strain specific. Most probably the studied probiotics act directly and/ or via modifications of the intestinal microbiota on the immune system (intraepithelial lymphocyte population).
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