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The objectives of our study were to enhance and evaluate an improved saponification method for the quantification of fatty acids (FA), with a specific focus on highly-unsaturated long-chain polyunsaturated FA (LPUFA) that are easily peroxidised in pig erythrocytes. These erythrocytes serve as a valuable large animal model for human studies. We implemented a modified saponification procedure, involving a shorter initial saponification at 95 °C for 2 min, followed by overnight saponification at 22--25 °C, and subsequent gentle base and acid-catalysed methylation of FA. These analytical procedures proved to be suitable for gas-chromatographic analysis of highly-unsaturated LPUFA, which are prone to peroxidation in processed piglet erythrocytes. The modified initial saponification at 95 °C, followed by overnight saponification at 22-25 °C, and subsequent methylation, were used to assess the suitability of erythrocytes for the determination of FA, tocopherol and malondialdehyde (MDA) concentrations in monogastric mammalian tissues. The content of docosahexaenoic acid (DHA) in erythrocytes can serve as a marker for DHA bioaccumulation in animal tissues and dietary contents of n-3LPUFA (especially DHA). The concentrations of δ-, γ-, α-tocopherols, total cholesterol and MDA increased in piglet erythrocytes during 16 days of exposure to dietary fat. The levels of FA and tocopherols can therefore be used in erythrocytes as indicators of the bioaccumulation yield of these compounds in animal tissues. Additionally, we propose that FA and tocopherol profiles in erythrocytes may be used as indicators of the concentrations of these components in diets and the effectiveness of their bioaccumulation in animal tissues. [ABSTRACT FROM AUTHOR]

The objective of the present study was to elucidate the effect of subclinical doses of Escherichia coli heat-labile enterotoxin (LT) on the antro-duodenal myoelectric activities of weaned pigs. Twelve weaned pigs were surgically implanted with three pairs of electrodes on the antrum 3 cm before the pylorus, 5 and 20 cm after the pylorus on the duodenum, respectively. An infusion cannula was inserted into the duodenum between duodenal electrodes. Using a wireless telemetry recording system, an electromyography (EMG) tracing lasting at least 24 h was recorded as the control, then another 24-h EMG recording was performed with a bolus intraduodenal infusion of LT (0.1 and 0.5 μg/kg b.w.). After a 1- to 2-day break, a 5-fold higher dose of LT was administered using the same protocol. In the antrum, LT administration barely modified the EMG signal. However, in the duodenum it prolonged the duration of phase II and the migrating myoelectric complex (MMC) cycle when compared with the control. The number of duodenal MMC cycles was also significantly diminished. Moreover, the migrating velocity of phase III was increased. The migrating action potential complex (MAPC) was present both without and with LT, but occurred more frequently following LT administration. In conclusion, LT caused a dose-dependent, lagged alteration in the duodenal MMC in weaned pigs, involving a reduction of the MMC number by lengthening phase II, increased phase III migration velocity, and increased MAPC frequency. The disturbances did not, however, result in diarrhoea and may reflect the induction of a local protection mechanism of the gut to expel unwanted foreign content from the lumen of the upper gut. [ABSTRACT FROM AUTHOR]

Protein maldigestion and malabsorption lead to malnutrition and are a feature of exocrine pancreatic insufficiency (EPI). Although it is the current standard, measurement of nitrogen in stool to assess protease activity is indirect. Up to 80% of hydrolysed proteins appear in blood in the form of peptides, so we developed a method to measure peptide-derived amino acids in plasma as a relevant measure of proteolysis, verified its accuracy, precision, and linearity, and validated it in a porcine model. We modified a ninhydrin method. Large proteins were eliminated from plasma with 10 kDa-cut-off centrifugal filters. Free and total amino acids were measured in permeate before and after its hydrolysis. Peptide-derived amino acids were quantified by subtracting free amino acids from total amino acids. We verified the method in vitro and by comparing results in healthy and EPI pigs. The accuracy of the analysis was close to 100%, with excellent precision (mean relative standard deviation for low, medium, and high amino acid levels = 0.88%) and with stringent linearity ( r 2  = 0.986, %RE = 5.23). The high-throughput ninhydrin method detected levels of peptide-derived amino acids in vivo with maximal changes seen approximately 2 hours postprandially in young pigs. The AUC and Cmax were significantly higher in healthy compared to EPI pigs ( P  = .0026 and P  = .0037, respectively). The high-throughput ninhydrin method is a sensitive, reliable, and practical method for the estimation of dietary peptide-derived amino acids. This assay endpoint could serve as a direct biomarker of protein digestion and absorption., (© The Author(s) 2024. Published by Oxford University Press.)

Preterm infants are most at risk of iron deficiency. However, our knowledge of the regulation of iron homeostasis in preterm infants is poor. The main goal of our research was to develop and validate an animal model of human prematurity to assess iron status in preterm infants. We performed a cesarean section on sows on the 109th day of pregnancy, which corresponds to the last trimester of human pregnancy. Preterm piglets showed decreased body weight, red blood cell indices, plasma iron level and transferrin saturation. Interestingly, higher hepatic and splenic non-heme iron content and plasma and hepatic ferritin levels were found in premature piglets compared with term ones. In addition, premature piglets showed higher mRNA levels of iron-regulatory hormone hepcidin in the liver than term animals, which have not been reflected in higher plasma hepcidin-25 levels. We also showed changes in hepcidin regulators, including hepatic bone morphogenetic protein 6, plasma erythroferrone and growth differentiation factor 15 in preterm piglets. Consequently, no difference was observed in iron-exporter ferroportin levels in the spleen and liver. Overall, it seems that premature piglets show a pattern of iron metabolism characteristic of functional iron deficiency and iron accumulation in the tissue.

Background: The possible existence of an acini-islet-acinar (AIA) reflex, involving mutual amylase and insulin interactions, was investigated in the current acute experiment on pigs., Aim: To confirm the existence of an AIA reflex and justify the placement of the exocrine and endocrine pancreatic components within the same organ., Methods: The study was performed on six pigs under general anesthesia. An intravenous glucose tolerance test was performed, with a bolus infusion of 50% glucose to the jugular vein, while amylase (5000 U/kg) or vehicle intrapancreatic infusions were administered via the pancreaticoduodenalis cranialis artery during 30 min with a 1 mL/min flow rate., Results: The amylase infusion to pancreatic arterial circulation inhibited and delayed the insulin release peak which is usually associated with the highest value of blood glucose and is typically observed at 15 min after glucose infusion, for > 1 h. The intrapancreatic infusion of the vehicle (saline) did not have any effect on the time frame of insulin release. Infusion of 1% bovine serum albumin changed the insulin release curve dramatically and prolonged the high range of insulin secretion, far beyond the glucose peak., Conclusion: Intrapancreatic arterial infusion of amylase interrupted the integrated glucose-insulin interactions. This confirms an AIA reflex and justifies placement of the exocrine and endocrine pancreatic components within the same organ., Competing Interests: Conflict-of-interest statement: The authors of this manuscript have the following competing interests: Stefan Pierzynowski and Kateryna Pierzynowska are the owners of Anara AB, Sweden. Piotr Wychowański is the owner of MED Sp. z o.o., Poland. Stefan Pierzynowski is the owner of Vitanano Sp. z o.o., Poland. Kamil Zaworski received funding from National Science Centre, Poland (Preludium, 2021/41/N/NZ9/01722). All other authors declare no competing interests., (©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.)

The purpose of this review was to analyze the scientific literature on exocrine pancreatic insufficiency (EPI) in dogs and cats and our own research on porcine model to compare animal- and microbial-derived enzymes in the treatment of animals with this disease. Clinical signs of EPI occur when more than 85% of the pancreatic parenchyma is non-functional. EPI can be a consequence of various diseases. The insufficient activity or deficiency of pancreatic enzymes leads to impaired digestion and absorption, and consequently, to malnutrition. The primary treatment for enzyme insufficiency is pancreatic enzyme replacement therapy (PERT). PERT in animals with EPI is a lifetime therapy. Most commercially available products are of animal origin (processed pancreata obtained from a slaughter house) and contain lipases, alpha-amylase, and proteases. Enzymes of microbial and plant origin seem to be a promising alternative to animal-derived enzymes, but to date there are no registered preparations containing all enzymes simultaneously for use in clinical practice to treat EPI. Results from some previous studies have highlighted the "extra-digestive" functions of pancreatic enzymes, as well as the actions of pancreatic-like microbial enzymes. For example, trypsin activates protease-activated receptor and provokes maturation of enterocytes and enterostatin inhibits fat absorption. It has been postulated that intrapancreatic amylase is the main component of the acini-islet-acinar axis-the reflex which down regulates insulin release, while gut and blood amylase exhibit anti-incretin actions "per se." Additionally, high but still physiological blood amylase activity coincide with physiological glucose homeostasis and a lack of obesity., (© 2024 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.)

The acini-islet-acinar (AIA) axis concept justifies the anatomical placement of the Langerhans islets within the exocrine pancreatic parenchyma and explains the existence of the pancreas as a single organ. Amylase has been suggested to play a key role as an anti-incretin factor. Oral glucose tolerance tests (OGTT) were performed on 18 piglets in both a healthy (prior to pancreatic duct ligation (PDL) surgery, study Day 10) and an exocrine pancreatic insufficient (EPI) state (30 days after PDL, study Day 48)). Amylase (4000 units/feeding) or Creon ® (100,000 units/feeding) was administered to pigs with the morning and evening meals, according to study design randomization, for 37 days following the first OGTT. Blood glucose levels, as well as plasma levels of insulin, GLP-1, and GIP, were measured, and the HOMA-IR index was calculated. EPI status did not affect the area under the curve (AUC) of insulin release, fasting insulin levels, or the HOMA-IR index, while amylase supplementation led to a significant ( p < 0.05) decrease in the above-mentioned parameters. At the same time, EPI led to a significant ( p < 0.05) increase in GLP-1 levels, and neither amylase nor Creon ® supplementation had any effects on this EPI-related increase. Fasting plasma levels of GIP were not affected by EPI; however, the GIP response in EPI and Amylase-treated EPI animals was significantly lower ( p < 0.05) when compared to that of the intact, healthy pigs. Orally administered amylase induces gut anti-incretin action, normalizing glucose homeostasis and reducing HOMA-IR as a long-term outcome, thus lowering the risk of diabetes type II development. Amylase has long-lasting anti-incretin effects, and one could consider the existence of a long-lasting gut memory for amylase, which decreases hyperinsulinemia and hyperglycemia for up to 16 h after the last exposure of the gut to amylase.

The aim of the study is to present the preliminary results of the in vivo application of Komagataeibacter xylinum E25 bacterial cellulose (BC) as a replacement material for produced defects during operations. Three pigs (sus scrofa domestica) had the same defects in the ear cartilage (4 × 4 cm) and in the rectus abdominis muscle (6 × 10 cm) with BC membranes implanted into them. The time of observation of the condition of the animals was 3 months. Implantation sites did not show clinical signs of complications in the form of inflammation or necrosis. Histologically, a normal scar was produced as a result of the material healing into the host's body. In one case, no residual implant material was found at the site of implantation, and the remodeled scar confirmed healing. No systemic inflammatory reaction was observed in any of the animals. The host organism's reaction to the bacterial cellulose allows us to believe that it meets the expectations as a material that can be widely used in reconstructive surgery. Nevertheless, this requires further research on a larger group and also using other foreign bodies. The next step would be an experiment on a group consisting of people.

Modern dental therapy makes use of prosthetic implant reconstructions, which are supported or retained on dental implants. The most frequent, long-term complications associated with these prosthetic implants include mucositis and peri-implantitis. Since mucositis is the initial inflammation of tissues supporting the dental implant, the management of this condition is thus crucial. The aim of the present study was to assess the effects of the placement of bioactive healing abutment for 48 h, in patients diagnosed with peri-implant mucositis. Moreover, the quantitative and qualitative shift in the bacterial profile of the biofilm present in the peri-implant pockets, was assessed by means of RT-PCR genotyping. Each patient was examined using a commercially available PET test protocol: the first sample was taken upon diagnosis (after which the bioactive healing abutment, with clindamycin at a dose of 30 mg, was used for 48 h and replaced with the prosthetic superstructure used so far by a patient); the second sample was taken two weeks after removal of the bioactive healing abutment. The effects of the intervention were clinically assessed using the PET test after the two weeks. A significant reduction in mucositis was observed following treatment, as measured by periodontal indices: modified Sulcus Bleeding Index—mBI (p < 0.001), modified Plaque Index—PLI (r = 0.69, Z= −4.43; p < 0.001) and probing depth—PD (Z = −4.61; p < 0.001). Significant differences in the occurrence of periopathogenic bacteria were also observed: Aggregatibacter actinomycetemcomitans (p < 0.014; Z = −2.45; r = 0.38), Treponema denticola (p < 0.005; Z = −2.83; r = 0.44), Tannerella forsythia (p < 0.001; Z = −4.47; r = 0.69) and Porphyromonas gingivalis (p < 0.132; Z = −1.51).

Butyrate, a by-product of gut bacteria fermentation as well as the digestion of fat in mother's milk, exerts a wide spectrum of beneficial effects in the gastrointestinal tissues. The present study aimed to determine the effects of sodium butyrate on small intestine contractility in neonatal piglets. Piglets were fed milk formula alone (group C) or milk formula supplemented with sodium butyrate (group B). After a 7-day treatment period, isometric recordings of whole-thickness segments of the duodenum and middle jejunum were obtained by electric field stimulation under the influence of increasing doses of Ach (acetylocholine) in the presence of TTX (tetrodotoxin) and atropine. Moreover, structural properties of the intestinal wall were assessed, together with the expression of cholinergic and muscarinic receptors (M1 and M2). In both intestinal segments (duodenum and middle jejunum), EFS (electric field stimulation) impulses resulted in increased contractility and amplitude of contractions in group B compared to group C. Additionally, exposure to dietary butyrate led to a significant increase in tunica muscularis thickness in the duodenum, while mitotic and apoptotic indices were increased in the middle jejunum. The expression of M1 and M2 receptors in the middle jejunum was significantly higher after butyrate treatment. The results indicate increased cholinergic signaling and small intestinal growth and renewal in response to feeding with milk formula enriched with sodium butyrate in neonatal piglets.

Background: The impact of concomitant obesity and sleep disorders on neuropeptides related to energy balance is poorly understood. The aim of this study was to assess the nocturnal profile of total ghrelin, obestatin, and leptin in patients with elevated BMI and to investigate the impact of breathing-related sleep disorders on these hormone levels., Methods: The study involved 58 patients with suspicion of obstructive sleep apnea (OSA). Patients underwent anthropometric and sleep examination and measurements of night ghrelin, leptin, and obestatin levels., Results: In patients with OSA ( n = 46), recognized on the basis of sleep examination outcomes, the correlation of anthropometric measurements with parameters of sleep disorders and ghrelin levels was observed, contrary to the control group ( n = 12). In the OSA group, levels of ghrelin were significantly lower than in the control group at 5:00 and 7:00. Levels of leptin in the OSA group were also lower than those in the control groups (not statistically significant). Profiles of obestatin in both groups were similar., Conclusions: Our results confirm the relationship between obesity and sleep-disordered breathing. Both these disorders affect ghrelin levels-parameters of obesity negatively correlate with hormone concentration, and OSA seems to lower ghrelin values in the second half of the night.