Immunodetection of selected pancreatic hormones under intragastric administration of apelin-13, a novel endogenous ligand for an angiotensin-like orphan G-protein coupled receptor, in unweaned rats.

Introduction: This study investigated the effects of intragastric administration of apelin-13 on the secretion of critical pancreatic hormones in a cohort of three-week-old Wistar rats. The research aimed to uncover apelin's modulatory roles in endocrine interactions dictating metabolic homeostasis during early life. Material and Methods: Rats were randomly assigned to control or experimental groups, receiving apelin-13 or saline for 14 days. The study population consisted of three-week-old Wistar rats of both sexes, weighing between 20 and 25 grams. Histological examination, analysis of variance and t-tests were employed to assess significant differences. Results: Distinctive alterations in large islet morphology were observed, indicating a notable reduction in size. Additionally, an increase in alpha- and beta-cell density within specific islet sizes was noted, suggesting significant changes in cell populations. The study found a substantial increase in mitotic activity and a decrease in apoptosis in small and medium-sized islets post apelin-13 administration, indicating its potential role in regulating cell survival and proliferation. Conclusion: The notable reduction in large islet size coupled with increased alpha and beta cell density implies a targeted impact of apelin-13 on pancreatic cell dynamics. Also, the observed increase in mitotic activity and decrease in apoptosis in small and medium-sized islets suggest its potential regulatory role in cell survival and proliferation within the pancreatic microenvironment.

Maternal acrylamide exposure changes intestinal epithelium, immunolocalization of leptin and ghrelin and their receptors, and gut barrier in weaned offspring.

Acrylamide (ACR) is an amide formed as a byproduct in many heat-processed starchy-rich foods. In utero ACR exposure has been associated with restricted fetal growth, but its effects of postnatal functional development of small intestine is completely unknown. The current study investigated the time- and segment-dependent effects of prenatal ACR exposure on morphological and functional development of small intestine in weaned rat offspring. Four groups of pregnant female Wistar rats were exposed to ACR (3 mg/kg b.w./day) for 0, 5, 10 and 15 days during pregnancy. Basal intestinal morphology, immunolocalization of gut hormones responsible for food intake and proteins of intestinal barrier, activity of the intestinal brush border disaccharidases, apoptosis and proliferation in intestinal mucosa were analyzed in offspring at weaning (postnatal day 21). The results showed that in utero ACR exposure disturbs offspring gut structural and functional postnatal development in a time- and segment-depended manner and even a short prenatal exposure to ACR resulted in changes in intestinal morphology, immunolocalization of leptin and ghrelin and their receptors, barrier function, activity of gut enzymes and upregulation of apoptosis and proliferation. In conclusion, prenatal ACR exposure disturbed the proper postnatal development of small intestine.

Effect of Adropin on Pancreas Exocrine Function in a Rat Model: A Preliminary Study.

The aim was to investigate the potential effect of adropin (ADR) on pancreatic−biliary juice (PBJ) secretion (volume, protein content, trypsin activity) in a rat model. The animals were divided into control and five experimental groups: adropin, CCK-8 (CCK-8 stimulation), capsaicin (capsaicin deactivation of afferents), vagotomy (vagotomy procedure), and vagal stimulation (vagal nerve stimulation). The experiment consisted of four phases, during which vehicle (0.9% NaCl) and three ADR boluses (5, 10, and 20 µg/kg BW) were administered i.v. every 30 min. PBJ samples were collected from each rat at 15 min intervals after boluses. Exogenous ADR failed to affect the pancreatic responses after vagotomy and the capsaicin pretreatment and reduced the PBJ volume, protein outputs, and trypsin activity in the adropin, CCK-8, and vagal stimulation groups in a dose-dependent manner. In all these groups, volume of PBJ was reduced only by the highest dose of ADR (p < 0.001 for adropin group and p < 0.01 for CCK-8 and vagal stimulation groups), and the protein outputs were reduced by the administration of ADR 10 µg/kg BW (adropin and CCK-8 groups, p < 0.01 in both cases) and 20 µg/kg BW (p < 0.001 for adropin and CCK-8 groups, p < 0.01 for vagal stimulation group). The 10 µg/kg BW dose of ADR reduced the trypsin output in the CCK-8 group (p < 0.01), and the highest ADR dose reduced the trypsin output in the CCK-8 (p < 0.001) and vagal stimulation (p < 0.01) groups. In conclusion, adropin in the analyzed doses exhibits the negative feedback pathway. This mechanism seems to participate in the regulation of pancreatic juice secretion via an indirect vagal mechanism.

Alterations in Small Intestine and Liver Morphology, Immunolocalization of Leptin, Ghrelin and Nesfatin-1 as Well as Immunoexpression of Tight Junction Proteins in Intestinal Mucosa after Gastrectomy in Rat Model.

The stomach is responsible for the processing of nutrients as well as for the secretion of various hormones which are involved in many activities throughout the gastrointestinal tract. Experimental adult male Wistar rats (n = 6) underwent a modified gastrectomy, while control rats (n = 6) were sham-operated. After six weeks, changes in small intestine (including histomorphometrical parameters of the enteric nervous plexuses) and liver morphology, immunolocalization of leptin, ghrelin and nesfatin-1 as well as proteins forming adherens and tight junctions (E-cadherin, zonula occludens-1, occludin, marvelD3) in intestinal mucosa were evaluated. A number of effects on small intestine morphology, enteric nervous system ganglia, hormones and proteins expression were found, showing intestinal enteroplasticity and neuroplasticity associated with changes in gastrointestinal tract condition. The functional changes in intestinal mucosa and the enteric nervous system could be responsible for the altered intestinal barrier and hormonal responses following gastrectomy. The results suggest that more complicated regulatory mechanisms than that of compensatory mucosal hypertrophy alone are involved.

Influence of Phytase Supplementation at Increasing Doses from 0 to 1500 FTU/kg on Growth Performance, Nutrient Digestibility, and Bone Status in Grower-Finisher Pigs Fed Phosphorus-Deficient Diets.

The objective of the current study is to assess the effects of the inclusion of 6-n phytase to a phosphorous-deficient diet on the growth performance (feed intake, average daily gain, and feed conversion ratio), apparent digestibility of calcium and phosphorus, and bone characteristics of grower-finisher pigs. The experimental diets included a phosphorus-deficient diet containing 0 (negative control), 250, 500, 1000, or 1500 FTU/kg of 6-phytase, and a diet formulated to meet the phosphorus nutrient requirements of pigs (positive control). Pigs were fed the experimental diets from the time they were ~35 kg body weight until they reached slaughter weight of ~110 kg. Bone status of the metacarpal (ash, mineral content) and femur (mineralization, geometry, and mechanical strength) bones were assessed. There was no effect of dietary treatment on feed intake. Feed conversion ratio was improved following inclusion of phytase at a dose of 500 FTU/kg or higher. Phytase inclusion at a dose of 1000 FTU/kg increased the average daily weight gain of grower-finisher pigs. Phytase inclusion at a dose of 500 FTU/kg was sufficient to increase metacarpal phosphorus content. Femur mid-diaphysis ash percentage was significantly increased even after the inclusion of the lowest dose of phytase. Analysis of structural parameters of femur mechanical strength (Young's modulus, yield stress, yield strain, ultimate stress, ultimate strain) showed that the inclusion of a phytase dose of 500 FTU/kg in growing/finishing diets was sufficient to significantly improve bone status of grower-finisher pigs at slaughter.

Effects of Yeast (Saccharomyces Cerevisiae) Probiotics Supplementation on Bone Quality Characteristics in Young Japanese Quail (Coturnix Japonica): The Role of Sex on the Action of the Gut-Bone Axis.

The aim of the study was to investigate the changes in bone geometry, histological structure, and selected mechanical characteristics in young male and female Japanese quails supplemented with Saccharomyces cerevisiae. Quails were fed a basal diet containing no yeast or a basal diet supplemented with 1.5% (15 g per 1 kg of diet) of inactive S. cerevisiae, for a period of 42 days. S. cerevisiae inclusion had no effect on bone weight, length, and density, diaphysis geometry (cross-sectional area, wall thickness, moment of inertia) or on the mechanical strength (yield load, ultimate load, stiffness, Young's modulus, yield stress, ultimate stress). Yeast supplementation improved the morphology of the articular cartilage both in male and female quails, as the total thickness of the articular cartilage was significantly increased. In trabecular bone, an increase in real bone volume and trabecular thickness was observed in females supplemented with S. cerevisiae, while in males the increase in trabecular number was accompanied by a reduction in trabecular thickness. The results of the present study demonstrate that S. cerevisiae, through a sex-dependent action on the gut-bone axis, improved the structure of articular cartilage and microarchitecture of trabecular bone. The positive effects of S. cerevisiae supplementation were more evident in female quails.

The effect of feed supplementation with Zakarpacki zeolite (clinoptilolite) on percentages of T and B lymphocytes and cytokine concentrations in poultry.

The available literature lacks information on the effect of Zakarpacki zeolite (clinoptilolite) on the immune system of poultry. Therefore, the aim of this study was to determine the effect of this zeolite on selected indicators of the immune response in poultry by evaluating the expression of cluster of differentiation (CD) surface molecules on T and B lymphocytes and the concentration of IL-2 and IL-10 in the blood. Ninety one-day-old Ross 308 broiler chicks were used in the study. The birds were divided into 3 groups of 30 each. The same basic diet was used in all groups, but groups II and III received a feed additive in the form of 2% and 3% zeolite. Blood samples were collected from all birds on the 40th day of observations. Weight gain in the birds in both experimental groups was significantly higher, and no clinical symptoms of disease were observed. The percentage of CD4+CD25+ T and B lymphocytes was higher in both groups receiving zeolite, but the percentage of CD8+CD25+ T lymphocytes was higher only in the group receiving 3% zeolite. There were no differences between the groups in the percentage of cells with CD3+ and MHC Class II expression. Higher serum concentrations of IL-2 and IL-10 were noted only in group III. The use of zeolites enhances antigen presentation and leads to increased Th1 and Th2 response. Excessive supply of zeolite in the feed leads to a local inflammatory response, which may cause damage to the intestinal barrier.

Apelin's effects on young rat gastrointestinal tract maturation.

Apelin is considered an important gut regulatory peptide with potential physiological roles in gastrointestinal cytoprotection and regulation of food intake and drinking behavior. The aim of this study was to determine the effects of intraperitoneal or intragastric apelin administration on gastric and intestinal epithelial apoptosis, mitosis and DNA repair enzyme 8-oxoguanine (OGG 1/2) expression in young Wistar rats (50±5 g b.wt.). Apelin-13 was intraperitoneally or intragastrically administered twice a day for 10 days (100 nmol/kg b.wt./2×day), and control groups received physiological saline as a placebo. The rats were sacrificed after treatment, and the gastric fundus, duodenum, middle jejunum and colon tissue samples were harvested for immunofluorescence studies. Intragastric administration of apelin-13 increased the apoptotic index in the stomach and colon tissues (P≤0.001) but decreased apoptosis in the duodenum and jejunum (P<0.001); this approach reduced the number of mitotic cells in the jejunum and colon but increased mitoses (P<0.001) in the duodenum. Finally, intragastric apelin-13 increased (P<0.001) OGG 1/2 enzyme expression in the stomach and jejunum and decreased its expression in the colon (P<0.01). However, intraperitoneal apelin-13 injection caused the opposite effect in the same regions of the gastrointestinal tract. In conclusion, apelin inhibits gastrointestinal tissue maturation in young rats, regardless of the administration route. However, further studies are required to clarify the mechanism of apelin action on gastrointestinal tract maturation in young rats.

Effect of apelin on mitosis, apoptosis and DNA repair enzyme OGG 1/2 expression in intestinal cell lines IEC-6 and Caco-2.

Apelin is a regulatory peptide, identified as an endogenous ligand of the Apelin receptor (APJ). Both the apelin and the APJ were detected in brain, lung, heart, mammary gland, kidney, placenta, adipose tissues and the gastrointestinal tract. Apelin is considered an important regulatory gut peptide with a potential physiological role in gastrointestinal cytoprotection, regulation of food intake and drinking behaviour. The aim of the present study was to assess the effect of the apelin on mitosis, apoptosis and the expression of DNA repair enzyme (OGG 1/2), and APJ receptor in intestinal cell lines: rat crypt (IEC-6) and human enterocyte model (Caco-2). The cell cultures were incubated with the apelin-12 (10-8 M) for 4, 6, 12, 24 and 48 h and the apoptosis (caspase 3), mitosis (Ki-67) and DNA repair enzyme (OGG1/2) markers were studied by Real-Time qRT-PCR and immunofluorescent methods. The results of Real-Time qRT-PCR and immunocytochemical analysis showed that the levels of mRNAs were inversely related to the expression level of corresponding proteins. Immunofluorescent studies revealed inhibitory effect of apelin-12 on apoptosis, mitosis and the expression of OGG1/2 in the intestinal crypt cell line IEC-6. However, in the enterocyte model Caco-2 cells apelin stimulated apoptosis and mitosis, and reduced OGG1/2 expression. These findings suggest that apelin may be involved in the control of epithelial cell turnover in the gastrointestinal tract.