Pancreatic Enzyme Activities Research Articles

8179 Complicated Pancreatitis Unveiled: A Case Of Untreated Primary Hyperparathyroidism

Abstract Disclosure: A. Rehan: None. H. Al Jumaili: None. J. Lim: None. Introduction: Acute pancreatitis (AP) triggered by hypercalcemia stemming from primary hyperparathyroidism (PHPT) is an infrequent occurrence, with a prevalence of about 1.5-7%. The proposed mechanism involves deposition of calcium in the pancreatic ducts and activation of pancreatic enzymes. While a causal connection between AP and PHPT has been described, it remains controversial. Calcium levels are thought to play a major role in determining the severity of the pancreatitis. Additionally, it is thought that genetic mutations may contribute to the development of AP in PHPT. Here, we report a case of necrotizing pancreatitis in a patient with untreated PHPT. Clinical Case: Patient is a 59-year-old male with a medical history of hypertension, nephrolithiasis and PHPT secondary to parathyroid hyperplasia (scheduled for parathyroidectomy) who presented with abdominal pain, nausea, vomiting and was found to have AP. He reported no history of alcohol use, trauma or insect bites. His workup was unrevealing for cholecystitis, choledocholithiasis or hypertriglyceridemia. He represented with calcium level of 16 mg/dL (NR 8.7-10.1) due to PHPT. PHPT was deemed the likely cause of AP. Course was further complicated by necrotizing pancreatitis and pseudocyst. A few months later, patient started having coffee ground emesis, worsening epigastric pain and weight loss of 60 lbs due to poor oral intake. CT abdomen/pelvis showed walled off pancreatic necrosis with peripancreatic stranding and concern for gastric outlet obstruction. EGD with PEG-J placement was done with plans for future endoscopic drainage. Few weeks after, he had another bout of worsening abdominal pain. Labs at the time showed corrected calcium between 11-13 mg/dL, PTH 147 pg/ml (NR 5-44), phosphorus 1.6 mg/dL (NR 2.5-4.5), vitamin D 25 hydroxy 19.5 ng/ml (NR 22-110). His condition became more complex with a hemorrhagic walled off pancreatic necrosis and splenic artery pseudoaneurysm requiring ICU care. The repeated escalation of symptoms resulted in multiple delays for the scheduled parathyroidectomy. Patient was started on treatment with Cinacalcet and vitamin D while in the hospital. Due to constraints in resources, he was unable to continue Cinacalcet outpatient. He subsequently was able to undergo parathyroidectomy with pathology showing benign adenoma. Conclusion: Hypercalcemic hyperparathyroidism causing AP, although rare, is often severe and poses a greater risk of complications making it important to be aware of this association, especially when more common causes have been excluded. Imaging techniques like cervical ultrasound, CT, and scintigraphy using 99mTc-Sestambi should be utilized to confirm suspicion. Surgical resection is the mainstay treatment, and having a multidisciplinary team with gastroenterologists, endocrinologists, and surgeons helps improve outcomes. Presentation: 6/2/2024

Prediction model establishment and validation for enteral nutrition aspiration during hospitalization in patients with acute pancreatitis.

Acute pancreatitis (AP) is a disease caused by abnormal activation of pancreatic enzymes and can lead to self-digestion of pancreatic tissues and dysfunction of other organs. Enteral nutrition plays a vital role in the treatment of AP because it can meet the nutritional needs of patients, promote the recovery of intestinal function, and maintain the barrier and immune functions of the intestine. However, the risk of aspiration during enteral nutrition is high; once aspiration occurs, it may cause serious complications, such as aspiration pneumonia, and suffocation, posing a threat to the patient's life. This study aims to establish and validate a prediction model for enteral nutrition aspiration during hospitalization in patients with AP. To establish and validate a predictive model for enteral nutrition aspiration during hospitalization in patients with AP. A retrospective review was conducted on 200 patients with AP admitted to Chengdu Shangjin Nanfu Hospital, West China Hospital of Sichuan University from January 2020 to February 2024. Clinical data were collected from the electronic medical record system. Patients were randomly divided into a validation group (n = 40) and a modeling group (n = 160) in a 1:4 ratio, matched with 200 patients from the same time period. The modeling group was further categorized into an aspiration group (n = 25) and a non-aspiration group (n = 175) based on the occurrence of enteral nutrition aspiration during hospitalization. Univariate and multivariate logistic regression analyses were performed to identify factors influencing enteral nutrition aspiration in patients with AP during hospitalization. A prediction model for enteral nutrition aspiration during hospitalization was constructed, and calibration curves were used for validation. Receiver operating characteristic curve analysis was conducted to evaluate the predictive value of the model. There was no statistically significant difference in general data between the validation and modeling groups (P > 0.05). The comparison of age, gender, body mass index, smoking history, hypertension history, and diabetes history showed no statistically significant difference between the two groups (P > 0.05). However, patient position, consciousness status, nutritional risk, Acute Physiology and Chronic Health Evaluation (APACHE-II) score, and length of nasogastric tube placement showed statistically significant differences (P < 0.05) between the two groups. Multivariate logistic regression analysis showed that patient position, consciousness status, nutritional risk, APACHE-II score, and length of nasogastric tube placement were independent factors influencing enteral nutrition aspiration in patients with AP during hospitalization (P < 0.05). These factors were incorporated into the prediction model, which showed good consistency between the predicted and actual risks, as indicated by calibration curves with slopes close to 1 in the training and validation sets. Receiver operating characteristic analysis revealed an area under the curve (AUC) of 0.926 (95%CI: 0.8889-0.9675) in the training set. The optimal cutoff value is 0.73, with a sensitivity of 88.4 and specificity of 85.2. In the validation set, the AUC of the model for predicting enteral nutrition aspiration in patients with AP patients during hospitalization was 0.902, with a standard error of 0.040 (95%CI: 0.8284-0.9858), and the best cutoff value was 0.73, with a sensitivity of 91.9 and specificity of 81.8. A prediction model for enteral nutrition aspiration during hospitalization in patients with AP was established and demonstrated high predictive value. Further clinical application of the model is warranted.

Incidence of Pancreatic Leakage in Isolated Biliary Limb Roux-en-Y Reconstruction in Whipple Procedure

Abstract Background Whipple is currently the procedure of choice in the treatment of localized pancreatic cancer. Isolated Roux-en-Y loop pancreaticojejunostomy was proposed as a method of reconstruction following pancreaticoduodenectomy. The idea is to isolate the bile from pancreatic secretions to minimize pancreatic enzyme activation and ultimately pancreatic fistula formation. It was also proposed that if a pancreatic fistula develops, it will be a pure fistula and thereby more benign as opposed to a mixed pancreatic fistula containing bile. Objective To estimate the incidence of pancreatic fistula in patients undergoing isolated Roux-en- Y hepaticojejunostomy reconstruction following pancreaticoduodenectomy. Patients and Methods In this prospective observational study, our aim is to calculate the incidence of post-operative pancreatic fistula in 20 patients that underwent Roux-en-Y isolated biliary limb reconstruction following pancreaticoduodenectomy for the treatment of pancreatic cancer. The study includes patients that underwent this procedure at Ain Shams University Hospitals in the period between January 2021 and July 2022 with a post-operative follow up period of 3 weeks. Patients were diagnosed with pancreatic cancer using MRI or CT scans with some of them also undergoing endoscopic US with biopsy. They were then admitted to one of the two main Ain Shams University Hospitals for pre-operative preparation. Results Our study did no estimate post-operative complications such as delayed gastric emptying and wound infection. Pancreaticoduodenectomy is a major surgery that carries a significant burden in terms of morbidity and mortality. Post-operative pancreatic fistula can be fatal as it can lead to vascular pseudoaneurysms that may fatally rupture. The idea behind using isolated biliary limb roux-en-y reconstruction is to alleviate the morbidity caused by post-operative pancreatic fistula. Conclusion Further comparative studies with a higher number of patients are recommended to assess and compare the value of isolated biliary limb reconstruction when compared to conventional reconstruction.

In vitro evaluation of algae and their effect as dietary ingredient on growth, chemical composition and intestinal functionality in juvenile turbot (Scophthalmus maximus)

The objectives of this study were to evaluate the in vitro protein hydrolysis of five algae (Arthrospira sp., Chlorella sp., Tisochrysis sp., Microchloropsis sp. and Ulva rigida) by the digestive proteases of Scophthalmus maximus, and to assess the effect of feeding turbot juveniles on diets supplemented with these algae in a 60-day feeding trial. The in vitro results revealed that algal protein was hydrolysed by turbot digestive proteases. Besides, the quantification of total amino acids released during the in vitro hydrolysis evidenced noticeable amino acid bioavailability, mainly in the case of Arthrospira sp. with up to 42.7% of the amino acids released after 120 min of in vitro hydrolysis. However, the structural characteristics of cell wall of each species influenced the protein accessibility as evidenced by the lower values obtained in Microchloropsis sp. For the in vivo feeding trial, 255 specimens of turbot, with an average initial weight of 26.4 ± 7.6 g, were used, and were fed with diets including 5% and 10% of a blend of the same microalgae tested in vitro (M5 and M10) or U. rigida (U5 and U10), compared to a control feed without algal biomass (CT). At the end of the feeding trial, growth performance and nutrient utilization did not show differences among the experimental groups. In addition, the fish fed with the diets supplemented with algae showed lower lipid oxidation in the muscle, and an improvement in digestive functionality, as the activity of pancreatic and intestinal enzymes increased. The ultrastructural study of the intestinal mucosa revealed an increase in the length of the microvilli and the absorptive surface in fish fed with the algae-supplemented diets, as well as decreased thickness of the lamina propria and the number of goblet cells. These results confirmed that the supplementation of turbot juvenile diets with low percentage of microalgae or U. rigida is appropriate for improving digestive functionality, as well as reducing the oxidation of lipids in the muscle.

Disrupted one-carbon metabolism in heifers negatively affects their health and physiology.

The objective of this study was to determine the dose-dependent response of one-carbon metabolite (OCM: methionine, choline, folate, and vitamin B12) supplementation on heifer dry matter intake on fixed gain, organ mass, hematology, cytokine concentration, pancreatic and jejunal enzyme activity, and muscle hydrogen peroxide production. Angus heifers (n = 30; body weight [BW] = 392.6 ± 12.6kg) were individually fed and assigned to one of five treatments: 0XNEG: total mixed ration (TMR) and saline injections at days 0 and 7 of the estrous cycle, 0XPOS: TMR, rumen-protected methionine (MET) fed at 0.08% of the diet dry matter, rumen-protected choline (CHOL) fed at 60g/d, and saline injections at days 0 and 7, 0.5X: TMR, MET, CHOL, 5-mg B12, and 80-mg folate injections at days 0 and 7, 1X: TMR, MET CHOL, 10-mg vitamin B12, and 160-mg folate at days 0 and 7, and 2X: TMR, MET, CHOL, 20-mg vitamin B12, and 320-mg folate at days 0 and 7. All heifers were estrus synchronized but not bred, and blood samples were collected on days 0, 7, and at slaughter (day 14) during which tissues were collected. By design, heifer ADG did not differ (P = 0.96). Spleen weight and uterine weight were affected cubically (P = 0.03) decreasing from 0XPOS to 0.5X. Ovarian weight decreased linearly (P < 0.01) with increasing folate and B12 injection. Hemoglobin and hematocrit percentage were decreased (P < 0.01) in the 0.5X treatment compared with all other treatments. Plasma glucose, histotroph protein, and pancreatic α-amylase were decreased (P ≤ 0.04) in the 0.5X treatment. Heifers on the 2X treatment had greater pancreatic α-amylase compared with 0XNEG and 0.5X treatment. Interleukin-6 in plasma tended (P = 0.08) to be greater in the 0XPOS heifers compared with all other treatments. Lastly, 0XPOS-treated heifers had reduced (P ≤ 0.07) hydrogen peroxide production in muscle compared with 0XNEG heifers. These data imply that while certain doses of OCM do not improve whole animal physiology, OCM supplementation doses that disrupt one-carbon metabolism, such as that of the 0.5X treatment, can induce a negative systemic response that results in negative effects in both the dam and the conceptus during early gestation. Therefore, it is necessary to simultaneously establish an optimal OCM dose that increases circulating concentrations for use by the dam and the conceptus, while avoiding potential negative side effects of a disruptive OCM, to evaluate the long-term impacts of OCM supplementation of offspring programming.

Structure-function relationships in (poly)phenol-enzyme binding: Direct inhibition of human salivary and pancreatic α-amylases

(Poly)phenols inhibit α-amylase by directly binding to the enzyme and/or by forming starch-polyphenol complexes. Conventional methods using starch as the substrate measure inhibition from both mechanisms, whereas the use of shorter oligosaccharides as substrates exclusively measures the direct interaction of (poly)phenols with the enzyme. In this study, using a chromatography-based method and a short oligosaccharide as the substrate, we investigated the detailed structural prerequisites for the direct inhibition of human salivary and pancreatic α-amylases by over 50 (poly)phenols from the (poly)phenol groups: flavonols, flavones, flavanones, flavan-3-ols, polymethoxyflavones, isoflavones, anthocyanidins and phenolic acids. Despite being structurally very similar (97% sequence homology), human salivary and pancreatic α-amylases were inhibited to different extents by the tested (poly)phenols. The most potent human salivary α-amylase inhibitors were luteolin and pelargonidin, while the methoxylated anthocyanidins, peonidin and petunidin, significantly blocked pancreatic enzyme activity. B-ring methoxylation of anthocyanidins increased inhibition against both human α-amylases while hydroxyl groups at C3 and B3′ acted antagonistically in human salivary inhibition. C4 carbonyl reduction, or the positive charge on the flavonoid structure, was the key structural feature for human pancreatic inhibition. B-ring glycosylation did not affect salivary enzyme inhibition, but increased pancreatic enzyme inhibition when compared to its corresponding aglycone. Overall, our findings indicate that the efficacy of interaction with human α-amylase is mainly influenced by the type and placement of functional groups rather than the number of hydroxyl groups and molecular weight.

A review: Pancreatic enzymes in the treatment of chronic pancreatic insufficiency in companion animals.

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.

Changes in Digestive Enzyme Activities during Larval Development of Spotted Seatrout (Cynoscion nebulosus)

The spotted seatrout (Cynoscion nebulosus)—an important commercial species—has a high potential for aquaculture in the Gulf of Mexico. To optimize its feeding during larval rearing, this study aims to evaluate the primary gastric (pepsin), intestinal (leucine aminopeptidase and alkaline phosphatase), and pancreatic (alkaline protease, trypsin, chymotrypsin, amylase, and lipase) enzyme activities from hatching to day 30. A multivariate analysis identified three digestive enzyme development stages during the spotted seatrout larval transformation. The first stage occurred between 1 (mean ± standard error (SE) = 1.73 ± 0.14 millimeter (mm) standard length (SL)) and 3 (2.14 ± 0.07 mm SL) days after hatching (DAH); a period of digestive stability showed the highest activity in amylase and bile salt-dependent lipase. The second stage (from 4 (2.53 ± 0.09 mm SL) to 20 (10.92 ± 0.51 mm SL) DAH) was a period of digestive transition, during which leucine aminopeptidase, chymotrypsin, and alkaline proteases were identified as the predominant enzymes from 4 to 5 DAH. In the third stage—a period of digestive stability—pepsin was the major enzyme that occurred between 25 (16.51 ± 0.81 mm SL) and 30 (25.91 ± 0.82 mm SL) DAH. These results indicate that the spotted seatrout larvae have a digestive system adapted to lipids and carbohydrates at the onset of feeding, with an immediate transition to protein digestion when exogenous feeding begins. Additionally, the digestive system of the spotted seatrout may be considered mature at 25 DAH. Further research is needed to elucidate the mechanisms of digestive tract development in the spotted seatrout larvae.

From Nature to Laboratory: The Impact of Leilem Leaves’ Ethanol Extract on Pancreatic Lipase Enzyme Activity

For many years, there have been theories on the possibility of preventing or delaying type 2 Diabetes Mellitus by altering some of its risk factors. Drugs that combat obesity have recently been researched concerning the prevention of type 2 Diabetes Mellitus. Inhibition of the digestive enzyme pancreatic lipase is a potential therapeutic strategy in treating and managing chronic diseases such as diabetes and obesity. Plants containing bioactive compounds are identified as potential sources of pancreatic lipase enzyme inhibitors. The use of natural compounds in inhibiting pancreatic lipase enzyme activity is considered to have more potential due to low toxicity and side effects. This study aims to determine the potential and activity effect of ethanol extract of leilem leaves on inhibiting pancreatic lipase enzyme. This study is a laboratory experimental study, the method of measuring lipase inhibition potential was performed using porcine lipase and PNPB with several modifications and using Orlistat as a positive control. Readings were taken using an ELISA reader at a wavelength of 405 nm. The data were then processed to obtain the IC50 value and relative potency. The results of in vitro studies have shown the potential of leilem leaf extract to inhibit pancreatic lipase enzyme activity. Qualitatively, the results showed that leilem leaf extract contains secondary metabolite compounds such as Alkaloids, Flavonoids, Tannins, Saponins, Steroids, and Terpenoids. Quantitatively, the results showed that the ethanol extract of leilem leaves had an absorbance value at the lowest concentration of 1.346 ± 0.53 and 0.709 ± 0.29 for the highest concentration. The IC50 result obtained was 137.89 μg/mL while the IC50 value of the positive control Orlistat was 77.022 μg/mL. Ethanol extract of leilem leaves (Clerodendrum minahassae Teijsm. &amp; Binn) has a potential value of 0.558.

Acidic solution irrigation as a novel approach for treating infected walled-off pancreatic necrosis

Walled-off pancreatic necrosis (WOPN) is a severe complication of acute pancreatitis arising from pancreatic or peripancreatic necrosis complicated by infection, leading to elevated mortality rates. Extravasation of pancreatic juice creates a weakly alkaline microenvironment favorable for the survivial of translocated intestinal bacteria and activated pancreatic enzyme-mediated autodigestion of pancreatic and peripancreatic tissues. Conventional treatments for WOPN involve invasive procedures with irrigation to mitigate postoperative infections. However, there remains a lack of a unified standard for lavage fluid. Herein, we raise the hypothesis that the utilization of acidic solution irrigation may serve as a promising strategy for preventing and treating infected WOPN. Acidic solutions create an unfavorable environment for translocated intestinal bacteria in WOPN cases. Additionally, the acidic microenvironment may alleviate self-digestion of pancreatic and peripancreatic tissues by inhibiting pancreatic enzyme activity. This innovative approach holds potential for enhancing the management and outcomes of infected WOPN.

The influence of obesity on the parameters of the kallikrein-kinin system and proteolysis in chronic biliary pancreatitis

Background. Clinical practice and science have accumulated data that obese patients suffer from severe forms of acute and chronic pancreatitis, which is explained by the accumulation of fat around the pancreas, a decreased activity of pancreatic enzymes. The purpose of the study is to describe the features of the kallikrein-kinin system and proteolysis in chronic biliary pancreatitis (CBP), depending on the presence of comorbid obesity. Materials and methods. One hundred and thirty-seven patients with chronic biliary pancreatitis were examined and divided into two groups depending on the presence of comorbid obesity: group I consisted of 22 patients with CBP and group II — of 115 patients with CBP and obesity. Results. The obtained results proved that an increase in body weight of patients with CBP lead to a more severe kallikrein-kinin system imbalance, with enhanced activation of inflammation and a decrease in the protective effect of the proteolysis. An increase in the degree of obesity in patients with CBP was accompanied by a more severe kallikrein-kinin system and proteolytic imbalance with an increase in the proteolytic enzymes level that have a damaging effect on the hepatic and pancreatic tissues and have a pro-inflammatory activity, as well as by a decrease in the content of the proteolytic enzyme inhibitors, which reliably weakened the protective effect of kallikrein-kinin system and proteolysis. Conclusions. 1) It was proved that there is a higher activity of the kallikrein-kinin system (accor­ding to proteolytic activation and kallikrein levels) and a decrease in the activity of proteolytic enzyme inhibitors (α2-macroglobulin and kininase II) in patients with chronic biliary pancreatitis and comorbid obesity compared to those without obesity (p &lt; 0.05). 2) An increase in the degree of obesity lead to an increase of proteolytic activity and a decrease in the content of the proteolytic enzyme inhibitors in patients with chronic biliary pancreatitis. It was proved the aggravating effect of obesity on the kallikrein-kinin system and proteolytic imbalance, which must be taken into account while forming a comprehensive treatment of such patients.

Antiobesity and antidiabetes effects of Cyperus rotundus rhizomes presenting protein tyrosine phosphatase, dipeptidyl peptidase 4, metabolic enzymes, stress oxidant and inflammation inhibitory potential

Diabetes is a significant global health concern that increases the vulnerability to various chronic illnesses. In view of this issue, the current research aimed to examine the effects of administering an extract derived from the tubers of Cyperus rotundus L (CrE) on obesity, type 1 diabetes, and liver-kidney toxicity. Through the utilization of HPLC-DAD analysis, it was discovered that the extract contained several components, including quercetin (47.8%), luteolin glucoside (17%), luteolin (7.56%), apigenin-7-glucoside (6.29%), naringinin (4.52%), and seven others. In vitro experiments they have demonstrated that CrE effectively inhibited key digestive enzymes associated with obesity and type 2 diabetes, such as DPP-4, PTP1B, lipase, and α-amylase, as evidenced by their respective IC50 values are about 23, 51,83, and 67 μg/ml respectively. Furthermore, when diabetic rats were administered CrE, the activity of pancreatic enzymes linked to inflammation, namely 5-lipoxygenase (5-LO), hyaluronidase (HAase), and myeloperoxidase (MPO), was significantly suppressed by 48, 41, 75, and 47%, respectively. Moreover, CrE exhibited protective effects on pancreatic β-cells by inhibiting the formation of thiobarbituric acid reactive substances (TBARS) by 65% and the induction of superoxide Dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) activities by 62, 108, and 112% respectively as compared to diabetic untreated rat. Additionally, CrE significantly inhibited the activities of intestinal, pancreatic, and serum lipase and α-amylase activities. In diabetic rats, CrE administration suppressed glycogen phosphorylase (GP) stimulated glycogen synthase (GS) activities by 45 and 30%; and this increased liver glycogen content by 45%. Furthermore, CrE modulated key hepatic enzymes involved in carbohydrate metabolism, including hexokinase (HK), glucose-6-phosphate dehydrogenase (G6PD), glucose-6-phosphatase (G6P), and fructose-1,6-bisphosphatase (FBP). Notably, the average food and water intake (AFI and AWI) of diabetic rats treated with CrE was reduced by 15 and 16% respectively as compared to those without any treatment. Therefore, this study demonstrated the effectiveness of Cyperus rotundus tubers in preventing and treating obesity and diabetes.

Influence of vasoactive intestinal polypeptide on growth performance, nutrient digestibility, nitrogen balance, and digestive enzyme activity in lambs.

This study evaluated if vasoactive intestinal polypeptide (VIP) influences growth performance, nutrient digestibility, nitrogen balance, and digestive enzyme activity. Sixteen wether lambs (69.6 ± 1.9kg) were housed in individual pens, adapted to a corn grain-based diet, and randomly assigned to 2 treatment groups. Lambs were injected intraperitoneally every other day for 28 d with saline (0.9% NaCl) containing no VIP (n = 8; control) or containing VIP (n = 8; 1.3 nmol/kg body weight [BW]). All lambs were transferred to individual metabolic crates for the final 7 d of the experiment to measure nitrogen balance and nutrient digestibility. At the end of the treatment period, lambs were slaughtered, and pancreatic tissue, small intestinal tissue, and rumen fluid were collected for protein, digestive enzymes, ruminal pH, and volatile fatty acid (VFA) analyses. Lambs treated with VIP had greater final BW, average daily gain, and gain:feed (P = 0.01, 0.05, 0.03, respectively). No differences between treatment groups were observed (P ≥ 0.25) for nutrient intake, digestibility, nitrogen retention, ruminal pH, and VFA concentrations. Moreover, VIP treatment did not influence (P ≥ 0.19) plasma glucose, urea N, and insulin concentrations. Treatment with VIP increased (P = 0.03) relative cecum weight (g/kg BW) and decreased (P = 0.05) relative brain weight. Pancreatic and intestinal digestive enzyme activities, except for duodenal maltase (P = 0.02), were not influenced (P ≥ 0.09) by VIP treatment. These data suggest that the administration of VIP may have potential to improve average daily gain and gain:feed in lambs fed grain-based diets.

MiR-605-3p may affect caerulein-induced ductal cell injury and pyroptosis in acute pancreatitis by targeting the DUOX2/NLRP3/NF-κB pathway.

Acute pancreatitis (AP) is a sudden-onset disease of the digestive system caused by abnormal activation of pancreatic enzymes. Dual oxidase 2 (DUOX2) has been found to be elevated in the progression of a variety of inflammatory diseases. Therefore, we analyzed the specific roles of DUOX2 in AP development. Blood samples were collected from of AP patients and healthy people, and the caerulein- stimulated human pancreatic duct cells (H6C7) were utilized to establish an AP cell model. Cell growth and apoptosis were measured using an MTT assay and TUNEL staining. Additionally, RT-qPCR and western blot assays were conducted to assess the RNA and protein expressions of the cells. ELISA kits were used to determine TNF-α, IL-6, IL-8, and IL-1β levels. The interaction between DUOX2 and miR-605-3p was predicted using the Targetscan database and confirmed by dual-luciferase report assay. We found that DUOX2 increased while miR-605-3p decreased in the blood of AP patients and caerulein-stimulated H6C7 cells. DUOX2 was targeted by miR-605-3p. Furthermore, DUOX2 knockdown or miR-605-3p overexpression promoted cell viability, decreased the TNF-α, IL-6, IL-8, and IL-1β levels, and inhibited apoptosis rate in caerulein-stimulated H6C7 cells. DUOX2 knockdown or miR-605-3p overexpression also increased the Bcl-2 protein levels and down-regulated Bax, cleaved-caspase-1, NLRP3 and p-p65. Interestingly, DUOX2 overexpression reversed the miR-605-3p mimic function in the caerulein-treated H6C7 cells. In conclusion, our research demonstrated that DUOX2 knockdown relieved the injury and inflammation in caerulein-stimulated H6C7 cells.

Thymol, a bioactive component of ocimum gratissimum linn. leaf ameliorated diabetes-induced exocrine pancreas insufficiency in male wistar rats

Background and aims: Exocrine pancreas insufficiency characterized by decreased pancreatic enzyme activity is one of the conditions associated with prolonged diabetes mellitus. Anti-diabetic activity of Ocimum gratissimum linn. leaf in experimental animals is well documented and using gas chromatography mass spectroscopy analysis, thymol has been identified as one of its bioactive components. This study investigated the effect of thymol on pancreatic alpha amylase activities and oxidative stress biomarkers in streptozotocin-induced diabetic rats. Methods: Twenty-four male Wistar rats grouped into four as Control, THY, DIA, and DIA+THY were used. Animals in DIA and DIA+THY were diabetic orally administered distilled water and thymol (20 mg/kg), respectively, while control and THY were normal rats treated similarly for 14 days. Fasting blood glucose (FBG), liver function, lipid profile, and oxidative stress biomarkers were determined in the blood while the pancreas was obtained for amylase activities. Results: Thymol treatment in the DIA+THY decreased FBG (119.75 ± 20.10 mg/dl), cholesterol (129.22 ± 8.34 mg/dl), triglycerides (116.41 ± 8.34 mg/dl), and low-density lipoprotein (101.89 ± 8.28 mg/dl) when compared with the DIA (209.25 ± 46.88 mg/dl, 156.31 ± 16.21 mg/dl, 143.13 ± 14.81 mg/dl, and 131.23 ± 13.38 mg/dl, respectively). Increased alanine transaminase and alkaline phosphatase observed in DIA were reversed in the Dia+THY. Pancreatic amylase activity was reduced in DIA (3.01 ± 0.50 Activity/mg.protein) compared with control (6.4 ± 0.42 Activity/mg.protein) and this was increased in DIA+THY (6.73 ± 0.39 Activity/mg.protein) relative to DIA. Increased plasma malondialdehyde and decreased catalase activity were observed in the DIA group and these were reversed in the DIA+THY group Conclusions: Thymol has hypoglycemic properties and stimulates pancreatic alpha-amylase activity through a mechanism that may be linked to its antioxidant properties.

Investigation of lipid digestion by fungal and porcine lipase supplements for severe pancreatic insufficiency using the in vitro dynamic TIM model

Exocrine pancreaticinsufficiency (EPI) is caused by insufficient secretion or limited activity ofpancreatic enzymes, especially lipase. Chronic pancreatitis (CP) is the mostcommon EPI disease. CP patients suffer from gastrointestinal symptoms such asdiarrhea, abdominal pain, and decreased uptake of nutrients as a consequence ofinadequate food digestion. To improve their nutritional status, pancreaticenzyme supplements can be orally administered, known as pancreatic enzymereplacement therapy (PERT). PERT is commonly based on oral porcinepreparations, despite their pH sensitivity which results in a limited timeframefor optimal activity. Instead, fungal enzymes could be used to prolong enzymeactivity duration, since they are active over a wide pH range. The performanceof fungal enzymes was compared to porcine enzymes by measuring total fattyacids under severe pancreatic insufficiency conditions in the tiny-TIMsgdynamic in vitro gastrointestinal model. Administration of 14.000 fungallipase units (Nortase) led to similar lipid digestion compared to 20.000porcine lipase units (Kreon and Pangrol). Additionally, opening the Nortasecapsules had no negative effect on fungal lipase activity. This workdemonstrates that fungal lipases, administered as capsules or powder, can beused as an alternative treatment for EPI patients.

CHARACTERISTICS OF PANCREATIC EXOCRINE FUNCTION IN PREMATURE INFANTS WITH PERINATAL PATHOLOGY

Introduction. According to the World Health Organization (WHO), the incidence of preterm birth is 10-15%, demonstrating with a consistent upward trajectory. Preterm infants exhibit immature and limited exocrine function of the pancreas during the first weeks of life that impedes enteral feeding and leads to the development of malnutrition and malabsorption.&#x0D; The aim of this study is to investigate the indicators of pancreatic exocrine function in premature infants with perinatal pathology. Materials and methods. The study included 91 premature infants with clinical manifestations of moderate and severe perinatal pathology with signs of disorders of the functional state of the digestive system (group I, gestational age 29 (0/7) - 36 (6/7) weeks) and 57 generally healthy newborns (group II, gestational age 35 (0/7) - 36 (6/7) weeks). The list of laboratory parameters characterizing the functional state of the pancreas in newborns includes: trypsin, amylase and lipase activity in blood serum. Statistical processing of data was performed using the software “STATISTICA” (StatSoft Inc., USA, version 10), program MedCalc (https://www.medcalc. org/index.php). Comparison of quantitative indicators with normal distribution was performed using Student’s t-test. The difference in parameters was considered statistically significant at p&lt;0.05. Results. Newborns of group I had clinical signs of food intolerance including weakened or absence of sucking reflex, regurgitation, stasis, intestinal paresis with delayed meconium and transitional stools, flatulence. Indicators of exocrine pancreatic function in newborns of the experimental group showed a decrease in the activity of trypsin, lipase, and amylase in blood serum compared with those in children of the control group. Conclusions. Clinical signs of dysfunction of digestive system in newborns are characterized by weakened or absence of sucking reflex, regurgitation, intestinal stasis and paresis, delayed passage of meconium and transitional stools, flatulence. Low activity of pancreatic enzymes in blood serum in the presence of gastrointestinal disorders confirms the primary exocrine pancreatic insufficiency, which impairs the assimilation of food ingredients in prematurely infants.

In vitro studies of magnesium oxide nano-particles of Saccharum officinarum root extract: Evaluation of the antidiabetic effect using alpha-glucosidase enzyme

Objective: Magnesium oxide nanoparticle green synthesis is said to have a wide range of biomedical uses. Inhibiting the activity of pancreatic enzymes is one method for treating diabetes. The goal of this study was to show in-vitro evidence that Saccharum officinarum magnesium oxide nanoparticles may inhibit the activity of the enzyme α-glucosidase. This study had a therapeutic focus as this could be emerging candidates of new drugs for controlling hyperglycemia. Materials and Procedures: Saccharum officinarum root extract was used to create magnesium oxide nanoparticles, which were then examined for bioactive components and tested for α-glucosidase inhibition. When the enzyme substrate solution was incubated with various doses of the grass-root Magnesium oxide nanoparticles, the activity of the enzyme was monitored to identify the kind of inhibition. Result: Magnesium oxide nanoparticles synthesized from Saccharum officinarum root extract showed concentration and time-dependent inhibition of α-glucosidase. An 85% inhibition was observed with 0.6 mg/ml of the synthesized nanoparticles. Conclusion: The results showed that the ethanolic extract of MgO nanoparticles synthesized by Saccharum officinarum exhibited effective antidiabetic activity, supporting its use in the treatment and control of diabetes.

Interleukin‑22 alleviates arginine‑induced pancreatic acinar cell injury via the regulation of intracellular vesicle transport system: Evidence from proteomic analysis.

Acute pancreatitis (AP) is a severe inflammatory condition characterized by the activation of pancreatic enzymes within acinar cells, leading to tissue damage and inflammation. Interleukin (IL)-22 is a potential therapeutic agent for AP owing to its anti-inflammatory properties and ability to promote tissue repair. The present study evaluated the differentially expressed proteins in arginine-induced pancreatic acinar cell injury following treatment with IL-22, and the possible mechanisms involved in IL-22-mediated alleviation of AP. AR42J cells were stimulated using L-arginine to establish an acinar cell injury model in vitro and the damaged cells were subsequently treated with IL-22. The characteristics of the model and the potential therapeutic effects of IL-22 were examined by CCK-8 assay, flow cytometry, TUNEL assay, transmission electron microscopy and ELISA. Differentially expressed proteins in cells induced by arginine and treated with IL-22 were assessed using liquid chromatography-mass spectrometry. The identified proteins were further subjected to Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis to elucidate their functional roles. The present study demonstrated that arginine-stimulated cells showed significant pathological changes resembling those in AP, which were alleviated after IL-22 treatment. Proteomic analysis then demonstrated that in IL-22-treated cells, proteins related to the formation and fusion of autophagosomes with lysosomes were significantly downregulated, whereas endocytosis related proteins were enriched in the upregulated proteins. After IL-22 treatment, western blotting demonstrated reduced expression of autophagy-associated proteins. In conclusion, by inhibiting the formation and fusion of autophagosomes with lysosomes, IL-22 may have mitigated premature trypsinogen activation, subsequently minimizing acinar cell injury induced by L-arginine. This was accompanied by concurrent upregulation of endocytosis, which serves a pivotal role in sustaining regular cellular material transport and signal propagation. This research underscored the potential of IL-22 in mitigating arginine-induced AR42J injury, which could be valuable in refining treatment strategies for AP.

Phytochemical analysis, in vitro and in silico effects from Alstonia boonei De Wild stem bark on selected digestive enzymes and adipogenesis in 3T3-L1 preadipocytes

BackgroundObesity is a global health issue arising from the unhealthy accumulation of fat. Medicinal plants such as Alstonia boonei stem bark has been reported to possess body weight reducing effect in obese rats. Thus, this study sought to investigate the in vitro and in silico effects of fractions from Alstonia boonei stem bark on selected obesity-related digestive enzymes and adipogenesis in 3T3-L1 preadipocytes.MethodTwo fractions were prepared from A. boonei: crude alkaloid fraction (CAF) and crude saponin fraction (CSF), and their phytochemical compounds were profiled using Liquid chromatography with tandem mass spectrometry (LCMS/MS). The fractions were assayed for inhibitory activity against lipase, α-amylase and α-glucosidase, likewise their antiadipogenic effect in 3T3-L1 adipocytes. The binding properties with the 3 enzymes were also assessed using in silico tools.ResultsEleven alkaloids and six saponin phytochemical compounds were identified in the CAF and CSF using LCMS/MS. The CAF and CSF revealed good inhibitory activity against pancreatic lipase enzyme, but weak and good activity against amylase respectively while only CSF had inhibitory activity against α-glucosidase. Both fractions showed antiadipogenic effect in the clearance of adipocytes and reduction of lipid content in 3T3-L1 adipocytes. The LCMS/MS identified compounds (41) from both fractions demonstrated good binding properties with the 3 enzymes, with at least the top ten compounds having higher binding energies than the reference inhibitors (acarbose and orlistat). The best two docked compounds to the three enzymes were firmly anchored in the substrate binding pockets of the enzymes. In a similar binding pattern as the reference acarbose, Estradiol-17-phenylpropionate (-11.0 kcal/mol) and 3α-O-trans-Feruloyl-2 α -hydroxy-12-ursen-28-oic acid (-10.0 kcal/mol) interacted with Asp197 a catalytic nucleophile of pancreatic amylase. Estradiol-17-phenylpropionate (-10.8 kcal/mol) and 10-Hydroxyyohimbine (-10.4 kcal/mol) interacted with the catalytic triad (Ser152-Asp176-His263) of pancreatic lipase while Estradiol-17-phenylpropionate (-10.1 kcal/mol) and 10-Hydroxyyohimbine (-9.9 kcal/mol) interacted with Asp616 and Asp518 the acid/base and nucleophilic residues of modelled α-glucosidase.ConclusionThe antiobesity effect of A. boonei was displayed by both the alkaloid and saponin fractions of the plant via inhibition of pancreatic lipase and adipogenesis.