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Biophysical Journal Feb 2003Earlier studies using atomic force microscopy (AFM) demonstrated the presence of fusion pores at the cell plasma membrane in a number of live secretory cells, revealing...
Earlier studies using atomic force microscopy (AFM) demonstrated the presence of fusion pores at the cell plasma membrane in a number of live secretory cells, revealing their morphology and dynamics at nm resolution and in real time. Fusion pores were stable structures at the cell plasma membrane where secretory vesicles dock and fuse to release vesicular contents. In the present study, transmission electron microscopy confirms the presence of fusion pores and reveals their detailed structure and association with membrane-bound secretory vesicles in pancreatic acinar cells. Immunochemical studies demonstrated that t-SNAREs, NSF, actin, vimentin, alpha-fodrin and the calcium channels alpha1c and beta3 are associated with the fusion complex. The localization and possible arrangement of SNAREs at the fusion pore are further demonstrated from combined AFM, immunoAFM, and electrophysiological measurements. These studies reveal the fusion pore or porosome to be a cup-shaped lipoprotein structure, the base of which has t-SNAREs and allows for docking and release of secretory products from membrane-bound vesicles.
Topics: Actins; Animals; Calcium Channels; Carrier Proteins; Cell Membrane; Cells, Cultured; Coated Pits, Cell-Membrane; Exocytosis; Male; Membrane Fusion; Membrane Proteins; Microfilament Proteins; Microscopy, Atomic Force; Microscopy, Electron; N-Ethylmaleimide-Sensitive Proteins; Pancrelipase; Qa-SNARE Proteins; Rats; Rats, Sprague-Dawley; Secretory Vesicles; Structure-Activity Relationship; Vesicular Transport Proteins; Vimentin
PubMed: 12547814
DOI: 10.1016/S0006-3495(03)74949-2 -
Surgical Case Reports Oct 2019Pancreatic exocrine insufficiency (PEI) is known to occur after total gastrectomy. We experienced a case of PEI occurring 18 years after surgery, leading to a...
BACKGROUND
Pancreatic exocrine insufficiency (PEI) is known to occur after total gastrectomy. We experienced a case of PEI occurring 18 years after surgery, leading to a potentially fatal condition of capillary leak syndrome (CLS).
CASE PRESENTATION
The case is a 58-year-old man on a healthy diet who underwent total gastrectomy 18 years before. He was admitted for a 3-month history of anasarca, steatorrhea, and hypoalbuminemia. An episode of fever occurred during workup, followed by pulmonary edema and shock. The patient was transferred to the intensive care unit and was started on fluid management with albumin infusion. A multidisciplinary team meeting was held, and a clinical diagnosis of PEI resulted in CLS was made and we started administration of oral pancrelipase to show clinical improvement. The patient was discharged, and he remained asymptomatic for 13 months.
CONCLUSION
In a post-gastrectomy patient with malnutrition, PEI should be suspected regardless of the period since surgery. When recognized, immediate replenishment of albumin and pancreatic enzymes should be initiated to prevent clinical deterioration.
PubMed: 31650279
DOI: 10.1186/s40792-019-0721-7 -
Internal Medicine (Tokyo, Japan) Jul 2022An 89-year-old woman underwent examinations for leg edema. Blood tests indicated low nutrition and low pancreatic enzymes, and a stool examination indicated fatty stool....
An 89-year-old woman underwent examinations for leg edema. Blood tests indicated low nutrition and low pancreatic enzymes, and a stool examination indicated fatty stool. Computed tomography showed pleural effusion, ascites, and cystic lesions in the pancreatic head and mural nodules within the cysts. Pancreatic juice cytology revealed adenocarcinoma. The diagnosis was pancreatic exocrine insufficiency caused by intraductal papillary mucinous carcinoma. The patient did not wish to undergo surgery. Therefore, diuretics, component nutrients, and pancreatic exocrine replacement therapy using pancrelipase were initiated. After starting treatment, her leg edema, pleural effusion, and ascites disappeared, and her activities of daily living improved markedly.
Topics: Activities of Daily Living; Adenocarcinoma, Mucinous; Adenocarcinoma, Papillary; Aged, 80 and over; Ascites; Carcinoma, Pancreatic Ductal; Edema; Exocrine Pancreatic Insufficiency; Female; Humans; Leg; Pancreatic Neoplasms; Pleural Effusion
PubMed: 34840231
DOI: 10.2169/internalmedicine.8611-21 -
International Journal of Molecular... Nov 2023The acini-islet-acinar (AIA) axis concept justifies the anatomical placement of the Langerhans islets within the exocrine pancreatic parenchyma and explains the...
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 ( < 0.05) decrease in the above-mentioned parameters. At the same time, EPI led to a significant ( < 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 ( < 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.
Topics: Animals; Swine; Incretins; Blood Glucose; alpha-Amylases; Pancrelipase; Insulin; Glucagon-Like Peptide 1; Amylases; Dietary Supplements; Gastric Inhibitory Polypeptide
PubMed: 38003366
DOI: 10.3390/ijms242216177 -
Caco-2 Cell Conditions Enabling Studies of Drug Absorption from Digestible Lipid-Based Formulations.Pharmaceutical Research Feb 2018To identify conditions allowing the use of cell-based models for studies of drug absorption during in vitro lipolysis of lipid-based formulations (LBFs).
PURPOSE
To identify conditions allowing the use of cell-based models for studies of drug absorption during in vitro lipolysis of lipid-based formulations (LBFs).
METHODS
Caco-2 was selected as the cell-based model system. Monolayer integrity was evaluated by measuring mannitol permeability after incubating Caco-2 cells in the presence of components available during lipolysis. Pure excipients and formulations representing the lipid formulation classification system (LFCS) were evaluated before and after digestion. Porcine mucin was evaluated for its capacity to protect the cell monolayer.
RESULTS
Most undigested formulations were compatible with the cells (II-LC, IIIB-LC, and IV) although some needed mucin to protect against damaging effects (II-MC, IIIB-MC, I-LC, and IIIA-LC). The pancreatic extract commonly used in digestion studies was incompatible with the cells but the Caco-2 monolayers could withstand immobilized recombinant lipase. Upon digestion, long chain formulations caused more damage to Caco-2 cells than their undigested counterparts whereas medium chain formulations showed better tolerability after digestion.
CONCLUSIONS
Most LBFs and components thereof (undigested and digested) are compatible with Caco-2 cells. Pancreatic enzyme is not tolerated by the cells but immobilized lipase can be used in combination with the cell monolayer. Mucin is beneficial for critical formulations and digestion products.
Topics: Administration, Oral; Caco-2 Cells; Drug Evaluation, Preclinical; Drug Liberation; Enzymes, Immobilized; Excipients; Fungal Proteins; Gastrointestinal Absorption; Humans; Lipase; Lipids; Lipolysis; Mucins; Pancrelipase; Pharmaceutical Preparations; Recombinant Proteins
PubMed: 29484506
DOI: 10.1007/s11095-017-2327-8 -
Journal of Cystic Fibrosis : Official... Dec 2009EUR-1008 (Zenpep [pancrelipase]) is a new, enteric-coated, porcine-derived pancreatic enzyme product (PEP) developed for the treatment of cystic fibrosis (CF) patients... (Clinical Trial)
Clinical Trial Randomized Controlled Trial
BACKGROUND
EUR-1008 (Zenpep [pancrelipase]) is a new, enteric-coated, porcine-derived pancreatic enzyme product (PEP) developed for the treatment of cystic fibrosis (CF) patients with malabsorption associated with exocrine pancreatic insufficiency (EPI). Unlike currently marketed PEPs, EUR-1008 contains the label-claimed lipase content. Safety and efficacy were assessed in younger (<7 years) and older (> or =7 years) CF patients with EPI.
METHODS
Two multicenter studies were conducted: a randomized, double-blind, placebo-controlled, crossover trial in patients > or =7 years of age (N=34) and a supplemental, open-label study in children <7 years of age (N=19). Use of any medications altering gastric pH/motility was prohibited during the studies. Outcome measures in the randomized trial included changes in the coefficient of fat absorption (CFA), coefficient of nitrogen absorption (CNA), and signs/symptoms of malabsorption for EUR-1008 vs. placebo. Outcome measures in the supplemental study included safety and response (defined as no steatorrhea and no overt signs/symptoms of malabsorption) to EUR-1008 vs. previous enzyme treatment.
RESULTS
In the randomized trial, EUR-1008 treatment compared to placebo resulted in a significantly higher mean CFA (88.3% vs. 62.8%, respectively) and CNA (87.2% vs. 65.7%, respectively) (both p<0.001) and reduced the incidence of malabsorption signs and symptoms in 32 evaluable patients. In the supplemental study, 11 of 19 patients met the criteria for responder with EUR-1008 at the end of the study vs. 10 of 19 patients at screening (previous PEP), and improvements in clinical symptoms were reported with EUR-1008 treatment. EUR-1008 was safe and well tolerated, and no serious drug-related AEs were reported in either study.
CONCLUSIONS
EUR-1008 was safe, well tolerated, and effective in CF patients of all ages with EPI-associated malabsorption in two clinical trials. Treatment led to clinically and statistically significant improvements in CFA and CNA in the randomized study, and control of malabsorption and clinical symptoms in both studies.
Topics: Adolescent; Child; Cholesterol; Cross-Over Studies; Cystic Fibrosis; Exocrine Pancreatic Insufficiency; Female; Humans; Malabsorption Syndromes; Male; Pancrelipase; Tablets, Enteric-Coated; Treatment Outcome; Vitamins; Young Adult
PubMed: 19683970
DOI: 10.1016/j.jcf.2009.07.006 -
Medical Oncology (Northwood, London,... Apr 2022To evaluate omega-3 fatty acid-rich enteral nutrient effects in patients with unresectable or recurrent biliary tract or pancreatic cancers during chemotherapy. Enteric...
Effects of an enteral nutrient-rich therapy with omega-3 fatty acids in patients with unresectable or recurrent biliary tract cancer or pancreatic cancer during chemotherapy: a case-control study.
To evaluate omega-3 fatty acid-rich enteral nutrient effects in patients with unresectable or recurrent biliary tract or pancreatic cancers during chemotherapy. Enteric nutritional supplements containing omega-3 fatty acids (Racol) was administered to aforementioned patients with cancers during chemotherapy. The skeletal muscle mass and blood test data were obtained pre-administration and 28 and 56 days after. Patients with pancreatic cancer were administered the digestive enzyme supplement pancrelipase (LipaCreon) 28 days after the start of Racol administration. The number of chemotherapies skipped due to neutropenia was recorded for 2 months before and after enteral nutrient initiation. In all 39 patients, the skeletal muscle mass increased on day 56 versus baseline (median 17.3 kg vs. 14.8 kg, p < 0.01), number of chemotherapies skipped decreased (mean: 0.65 times/month vs. 1.3 times/month, p = 0.03), and retinol-binding protein (mean: 2.56 mg/dL vs. 2.42 mg/dL, p = 0.05) increased. Patients with pancreatic cancer showed increased blood eicosapentaenoic acid concentration on day 56 versus baseline (median: 48.1 μg/mL vs. 37.0 μg/mL, p = 0.04) and increased skeletal muscle mass (median 16.8 kg vs. 14.4 kg, p = 0.006). Baseline median neutrophil count increased significantly from 2200/μL at baseline to 2500/μL (p = 0.04). Patients with biliary tract cancer during chemotherapy also exhibited increased skeletal muscle mass following omega-3 supplementation (median 17.3 kg vs. 15.8 kg, p = 0.01). In patients undergoing chemotherapy for unresectable or post-recurrence pancreatic and biliary tract cancers, high-omega-3 fatty acid nutrition therapy use improved skeletal muscle maintenance and chemotherapy dosing intensity.
Topics: Biliary Tract Neoplasms; Case-Control Studies; Fatty Acids, Omega-3; Gastrointestinal Neoplasms; Humans; Nutrients; Pancreatic Neoplasms
PubMed: 35478069
DOI: 10.1007/s12032-021-01625-4 -
Transplant International : Official... Sep 2021Static cold storage (SCS) is the standard method for pancreas preservation prior to transplantation; however, it does not permit organ assessment. Normothermic...
Static cold storage (SCS) is the standard method for pancreas preservation prior to transplantation; however, it does not permit organ assessment. Normothermic reperfusion (NR) is utilized clinically for other organs to assess viability. Our aim was to develop NR using normothermic machine perfusion technique to simulate reperfusion at the time of transplantation, enabling evaluation of oxygenated hypothermic machine perfusion (HMPO2) as a newer strategy to optimize pancreas preservation. 13 porcine pancreases procured after circulatory death were divided into 3 groups: 4 pancreases preserved using SCS, and 2 groups preserved by HMPO2 (n = 4 and n = 5, differing by type of preservation solution). Duration of perfusion or cold storage was 6 hours before the 1-hour assessment using NR. Outcome measures were perfusion characteristics, biochemistry and change in tissue water mass as oedema assessment. During NR, the HMPO2 groups demonstrated better perfusion characteristics, normal macroscopic appearances, decreased water mass and one HMPO2 group demonstrated a response to glucose stimulation. Conversely, the SCS group showed an increased water mass and developed early macroscopic appearances of oedema, interstitial haemorrhage and minimal portal outflow. This study suggests that ex situ assessment of pancreases by NR is promising, and that HMPO2 may be better than SCS.
Topics: Animals; Organ Preservation; Pancreas; Pancrelipase; Perfusion; Reperfusion; Swine
PubMed: 34448276
DOI: 10.1111/tri.13990 -
Antimicrobial Agents and Chemotherapy Jul 2017Tenofovir disoproxil fumarate (TDF), a prodrug of tenofovir, has oral bioavailability (25%) limited by intestinal transport (P-glycoprotein), and intestinal degradation...
Tenofovir disoproxil fumarate (TDF), a prodrug of tenofovir, has oral bioavailability (25%) limited by intestinal transport (P-glycoprotein), and intestinal degradation (carboxylesterase). However, the influence of luminal pancreatic enzymes is not fully understood. Physiologically based pharmacokinetic (PBPK) modeling has utility for estimating drug exposure from data. This study aimed to develop a PBPK model that included luminal enzyme activity to inform dose reduction strategies. TDF and tenofovir stability in porcine pancrelipase concentrations was assessed (0, 0.48, 4.8, 48, and 480 U/ml of lipase; 1 mM TDF; 37°C; 0 to 30 min). Samples were analyzed using mass spectrometry. TDF stability and permeation data allowed calculation of absorption rates within a human PBPK model to predict plasma exposure following 6 days of once-daily dosing with 300 mg of TDF. Regional absorption of drug was simulated across gut segments. TDF was degraded by pancrelipase (half-lives of 0.07 and 0.62 h using 480 and 48 U/ml, respectively). Previously reported maximum concentration (; 335 ng/ml), time to (; 2.4 h), area under the concentration-time curve from 0 to 24 h (AUC; 3,045 ng · h/ml), and concentration at 24 h (; 48.3 ng/ml) were all within a 0.5-fold difference from the simulated (238 ng/ml), (3 h), AUC (3,036 ng · h/ml), and (42.7 ng/ml). Simulated TDF absorption was higher in duodenum and jejunum than in ileum (p<0.05). These data support that TDF absorption is limited by the action of intestinal lipases. Our results suggest that bioavailability may be improved by protection of drug from intestinal transporters and enzymes, for example, by coadministration of enzyme-inhibiting agents or nanoformulation strategies.
Topics: ATP Binding Cassette Transporter, Subfamily B, Member 1; Adolescent; Adult; Anti-HIV Agents; Carboxylesterase; HIV Infections; Humans; Lipase; Male; Middle Aged; Pancrelipase; Tenofovir; Young Adult
PubMed: 28416547
DOI: 10.1128/AAC.00105-17 -
The Journal of Pediatric Pharmacology... Apr 2007OBJECTIVE Pancreatic enzyme products were available before the 1938 passage of the Federal Food, Drug, and Cosmetic Act and have to date been marketed without required...
In Vitro Comparison of Physical Parameters, Enzyme Activity, Acid Resistance, and pH Dissolution Characteristics of Enteric-Coated Pancreatic Enzyme Preparations: Implications for Clinical Variability and Pharmacy Substitution.
OBJECTIVE Pancreatic enzyme products were available before the 1938 passage of the Federal Food, Drug, and Cosmetic Act and have to date been marketed without required safety and efficacy testing. Despite a lack of demonstrated bioequivalence, they are often substituted for each other without physician or patient consent or monitoring. We investigated the in vitro variability of key performance parameters among a representative group of currently available pancreatic enzyme formulations.MATERIALS AND METHODS Three "branded" preparations (Creon 20 Minimicrospheres, Pancrease MT 20, Ultrase MT 20) and 3 "generic" formulations (Pangestyme CN-20, Pancrelipase 20,000 URL, and Lipram CR 20) were evaluated in vitro for physical parameters of the capsules, actual vs. labeled enzyme activity, resistance of the enteric coating to simulated gastric acid, and kinetics of simulated duodenal lipase release. All products were labeled as providing 20,000 units of lipase activity per capsule.RESULTS All products varied considerably in the percentage relationship between actual and labeled lipase activity. Actual lipase activity exceeded 165% of the label claim in 4 batches of the Pangestyme product and 1 batch of the Lipram product. All batches of the Creon, Lipram, Ultrase, and Pancrease products were found to have residual lipase activity above 80% of their baseline measurements after testing in simulated gastric acid; residual lipase activity varied significantly among batches of the Pangestyme product and was only 1% for the Pancrelipase product. The Creon and Lipram products demonstrated effective protection by the enteric coating at pH <6.0 and rapid release of enzymatic activity at pH ≥6.0. The Pangestyme and Pancrelipase products showed substantial activity of released enzymes already at pH 5.0. Release kinetics were inconsistent between batches for the Ultrase and Pancrease products.CONCLUSION This study confirms the existence of "branded"-to-"generic," product-to-product, and batch-to-batch variability among representative pancreatic enzyme formulations with pharmaceutically equivalent labels. The results confirm current cautions regarding pharmacy substitution of pancreatic enzyme products and support the announcement by the US Food and Drug Administration, made subsequent to this study, that as of April 2008 approved new drug applications will be required in order to ensure the quality, potency, and stability of these products.
PubMed: 23055848
DOI: 10.5863/1551-6776-12.2.115