Histopathological Examination Of Pancreatic Tissues Research Articles

Evaluation of antidiabetic and antioxidant activity of methanolic extract of Limonia acidissima leaves in alloxan-induced diabetic wister albino rats: a randomized controlled experimental study

Objectives: The present study was carried out to find graded-doses antidiabetic and antioxidant activities of methanolic extract of Limonia acidissima (MELA) leaves in rats. Materials and Methods: Male Wistar rats were randomly divided into 6 groups (n=8): Normal control, Diabetic control (Alloxan-induced), and test groups where varying doses of MELA (100mg/kg, 200mg/kg, and 400mg/kg) or Metformin (100mg/kg) administered post-alloxan induction. Random blood sugar (RBS) levels were estimated at specific intervals over a 28-day period. Moreover, antioxidant markers (malondialdehyde, superoxide dismutase, and reduced glutathione) were assessed on day 28. Histopathological examination of pancreatic tissue was conducted. Results: Overall, mean RBS levels decreased at all intervals. There was a significant decrease in median malondialdehyde, an increase in median superoxide dismutase, and reduced glutathione levels (all p-values<0.0001). The change in RBS (p-values<0.0001) and antioxidants (p-value>0.05) levels was greater in Group VI than in Group V. On histopathology, Groups VI and V demonstrated regeneration of necrotic β-cells in the islets, and this was more pronounced than Group IV. Conclusion: Results of our study suggested that MELA possesses dose-dependent significant antidiabetic and antioxidant activity.

Bone marrow stem cell-derived β-cells: New issue for diabetes cell therapy

This investigation aimed to establish the promising role of insulin-producing cells (IPCs) growing from bone marrow-mesenchymal stem cells (BM-MSCs) in relieving hyperglycemia induced in rats. BM-MSCs were differentiated into IPCs using three different protocols. The efficiency of BM-MSCs differentiation into IPCs in vitro was confirmed by detecting IPCs specific gene expression (Foxa-2, PDX-1 and Ngn-3) and insulin release assay. The in vivo study design included 3 groups of male Wistar rats; negative control group, diabetic group and IPCs-transfused group (5 ×106 cells of the most functional IPCs/rat). One month after IPCs infusion, serum glucose, insulin, c-peptide and visfatin levels as well as pancreatic glucagon level were quantified. Gene expression analysis of pancreatic Foxa-2 and Sox-17, IGF-1 and FGF-10 was done. Additionally, histological investigation of pancreatic tissue sections was performed. Our data clarified that, the most functional IPCs are those generated from BM-MSCs using differentiation protocol 3 as indicated by the significant up-regulation of Foxa-2, PDX-1 and Ngn-3 gene expression levels. These findings were further emphasized by releasing of a significant amount of insulin in response to glucose load. The transplantation of the IPCs in diabetic rats elicited significant decline in serum glucose, visfatin and pancreatic glucagon levels along with significant rise in serum insulin and c-peptide levels. Moreover, it triggered significant up-regulation in the expression levels of pancreatic Foxa-2, Sox-17, IGF-1 and FGF-10 genes versus the untreated diabetic counterpart. The histopathological examination of pancreatic tissue almost assisted the biochemical and molecular genetic analyses. These results disclose that the cell therapy holds potential to develop a new cure for DM based on the capability of BM-MSCs to generate β-cell phenotype using specific protocol.

Pancreatic Pathological Changes in Murine Toxoplasmosis and Possible Association with Diabetes Mellitus.

Previous studies have reported involvement of Toxoplasma gondii (T. gondii) infections in the pathogenesis of some autoimmune diseases, such as polymyositis, rheumatoid arthritis, autoimmune thyroiditis, and Crohn's disease. However, data on the association between T. gondii infections and Type 1 diabetes mellitus (T1DM) are still controversial. Therefore, in the present study, we aimed to investigate the pancreatic pathological changes in mouse models with acute and chronic toxoplasmosis and their association with T1DM. Three groups (10 mice each) of male Swiss Albino mice were used. One group of mice was left uninfected, whereas the second and third groups were infected with the acute virulent T. gondii RH strain and the chronic less virulent Me49 T. gondii strain, respectively. T. gondii-induced pancreatic pathological changes were evaluated by histopathological examination of pancreatic tissues. Moreover, the expression of insulin, levels of caspase-3, and the pancreatic infiltration of CD8+ T cells were evaluated using immunohistochemical staining. Pancreatic tissues of T. gondii-infected animals showed significant pathological alterations and variable degrees of insulitis. Mice with acute toxoplasmosis exhibited marked enlargement and reduced numbers of islets of Langerhans. However, mice with chronic toxoplasmosis showed considerable reduction in size and number of islets of Langerhans. Moreover, insulin staining revealed significant reduction in β cell numbers, whereas caspase-3 staining showed induced apoptosis in islets of Langerhans of acute toxoplasmosis and chronic toxoplasmosis mice compared to uninfected mice. We detected infiltration of CD8+ T cells only in islets of Langerhans of mice with chronic toxoplasmosis. Acute and chronic toxoplasmosis mice displayed marked pancreatic pathological changes with reduced numbers of islets of Langerhans and insulin-producing-β cells. Since damage of β cells of islets of Langerhans is associated with the development of T1DM, our findings may support a link between T. gondii infections and the development of T1DM.

Coriander Oil Reverses Dexamethasone-Induced Insulin Resistance in Rats.

In the present study, we aimed to investigate the effect of coriander oil on dexamethasone-induced insulin resistance in rats and characterize its chemical composition using gas chromatography-mass spectrometry (GC-MS). Rats were divided into five groups (n = 6): Normal control, insulin resistance (IR) control, IR + metformin (50 mg/kg/day, PO, Per Oral), IR + coriander oil low dose (0.5 mL/kg, PO), and IR + coriander oil high dose (1 mL/kg, PO). IR groups were injected with a dose of 10 mg/kg dexamethasone subcutaneously for four consecutive days. All groups received either vehicle or drugs daily for four days. Animal weights and pancreatic weights were measured, and oral glucose tolerance test was performed at the end of study. Fasting glucose, triglycerides (TG), total cholesterol (TC), HDL and insulin levels in serum, MDA, and GSH levels in pancreatic tissue were measured and HOMA-IR was calculated. Immunoexpression of apoptosis markers BAX, and BCL2 was measured in pancreatic tissues and BAX/BCL2 ratio was calculated. Histopathological examination of pancreatic tissues was also performed. Pancreatic weight, serum HDL, pancreatic GSH, and BCL2 were decreased while serum glucose, insulin, TG, TC levels, AUC of OGGT, HOMA-IR, pancreatic MDA, BAX, and BAX/BCL2 ratio were increased in IR rats. Histopathological examination showed congestion, vacuolation and hemorrhage in pancreatic islets. These changes were reversed by metformin and the high dose of coriander oil treatments. The obtained activities could be attributed to the presence of 21 volatile compounds, identified by GC-MS. Our study indicates that coriander oil can be used as an adjuvant antihyperglycemic agent in type 2 diabetes. Further experiments are needed to determine the therapeutic dose and the treatment time.

Ethyl Acetate Fraction of Total Phenols from Zanthoxylum zanthoxyloides Leaves Treated Alloxan-Induced Diabetic Albino Rats

Aims: Ethyl acetate fraction of total phenols from Zanthoxylum zanthoxyloides leaves was evaluated for its effects in alloxan-induced diabetic male albino rats.
 Methodology: Diabetes was induced in rats by administrating alloxan monohydrate at a dose of 130 mg/kg body weight. The ethyl acetate fraction was administered to rats at 100, 200 and 400 mg/kg b.w. daily for 15 days. Blood glucose, lipid profile and pancreatic histopathology were monitored using standard methods.
 Results: Results indicated that two weeks after treatment with Z. zanthoxyloides, significant (p<0.05) reductions in hyperglycemic blood glucose, total cholesterol and low-density lipoproteins (LDL) were recorded; with lowest values recorded in the group treated with the highest concentration of the extract. Histopathological examination of pancreatic tissues supported the protective effect of Z. zanthoxyloides phenol extract against diabetic damage.
 Conclusion: In conclusion, total phenols of Z. zanthoxyloides exerted antidiabetic effects in alloxan-induced diabetic rats and protected pancreatic tissues from diabetic damage.

Anti- diabetic activity of new Synthesized Flavonoid compound

Background: Diabetes mellitus (DM) is associated with long-term damage, dysfunction, of various organs. Objectives: Study aims to assess the role of new Synthesized Flavonoid compound on experimentally induced diabetes. Methods: 50 adult male albino rats divided into 5 groups. Group 1 (control group, rats were orally administered with 1 ml saline daily). Group 2 (DMSO group, rats were orally administered with 0.2 % DMSO for 60 day orally). Group 3 (positive control, animals were injected intraperitoneally with 60 mg/kg b.wt streptozotocin followed by intraperitoneal injection with 120 mg/kg b.wt of Nicotinamide after 15 minute).Group 4 (standard group, diabetic animals treated with 100 mg/kg b.wt of metformin for 60 day orally). Group 5 (therapeutic group, diabetic rats treated with 50 mg /kg b.wt of Ethyl 2-amino-4-phenyl-4H-benzo(h)chromene-3-carboxylate for 60 day orally). At the end of experimental period blood serum & plasma samples, liver, kidney and pancreatic tissues were collected. Results: diabetic rats showed significant increase in plasma glucose, serum urea, creatinine, cholesterol and triglyceride . Also significant increase in mean level of Fetuin A and Netrin-1 in serum and different organs (Liver, kidney, Pancreas) in compared to control group. Oral administration of Ethyl 2-amino-4-phenyl-4H-benzo(h)chromene-3-carboxylate cause decrease in elevated biochemical parameters. Also, decrease Fetuin A and Netrin-1 levels when compared with diabetic rats. Molecular docking studies confirmed binding of compound with Fetuin A and Netrin-1 proteins in terms of energy and revealed of the existence of hydrogen bond ,hydrophobic interaction,Our results were confirmed by histopathological examination of pancreatic tissue. Conclusion: this study suggests that Ethyl 2-amino-4-phenyl-4H-benzo(h)chromene-3-carboxylate exihibits antihyperglycemic activity in streptozotocin- induced diabetic rats.

Antidiabetic activity of thin film containing astaxanthin-loaded nanoemulsion using carboxymethylcellulose sodium polymer on alloxan-induced diabetic rabbit.

The present study was conducted to evaluate the potency of thin film containing astaxanthin-loaded nanoemulsion (FDT-As-NE) in lowering blood glucose levels on alloxan-induced diabetic rabbit (ADR). Astaxanthin nanoemulsion (As-NE) was prepared using self-nanoemulsifying method, followed by incorporated into the carboxymethylcellulose sodium matrix polymer using a solvent casting method to form a thin film. The evaluation of FDT-As-NE was performed by chemical, physical, and mechanical characterizations. The administration of thin film was done by an intraoral route. New Zealand albino rabbits were induced with alloxan to get experimental diabetic animals. The antidiabetic activity was carried out in three groups of treatment. Group I was ADR treated by FDT-As-NE, Group II was ADR treated by pure astaxanthin, while Group III was normal control. The measurement of fasting means blood glucose levels was carried out in 0 days (before treatment) and after 14 days of treatment. The histopathological analysis of the pancreas was also examined. Data were statistically evaluated using Kruskal–Wallis statistical test. P < 0.05 was considered statistically significant. FDT-As-NE had good physical and mechanical characteristics that suitable for intraoral administration. Group I reduced elevated blood glucose levels compared to Group II (P < 0.01). Histopathological examination of pancreatic tissue for a Group I showed the normal condition of pancreatic β-cell, suggesting the absence of any pathological lesions. These results suggest that thin film containing astaxanthin-loaded nanoemulsion administered by an intraoral route potentially useful for reducing glucose levels.

Eugenia jambolana extract reduces the systemic exposure of Sitagliptin and improves conditions associated with diabetes: A pharmacokinetic and a pharmacodynamic herb-drug interaction study

Eugenia jambolana (EJ) is an Indian traditional herb widely used for the treatment of diabetes mellitus. This herb is globally marketed as single or multi herb formulations. Many diabetes patients consume EJ extract oral hypoglycemic drugs together. This calls for a need to assess risks versus benefit of this co-administration.In present investigation, pharmacodynamic and pharmacokinetic interactions of aqueous extract of EJ seeds at the dose of 400 mg/kg are studied with 10 mg/kg of oral hypoglycaemic drug sitagliptin (SITA) by co-administrating them for 28 days in streptozotocin (STZ) induced diabetic rats. The pharmacokinetic parameters of SITA were determined using HPLC-ESI-MS/MS and it was found that the combination treatment reduces the systemic exposure of SITA by showing 38.70% reduction in concentration maximum (Cmax) and 22.40% reduction in area under curve (AUC). Despite low levels of SITA, the combination demonstrated a significant reduction in blood glucose level when compared with individual drug and individual extract administered groups during pharmacodynamic study. In addition, the liver function, the kidney function and the lipid parameters were found to be significantly improved and beneficial effects were found with respect to food intake and water intake and urine output in case of combination treatment groups when compared with individual treatment groups. Histopathological examination of pancreatic tissue suggests its significant recovery of having normal acinus with better cell protection in combination treatment. In conclusion, the combination treatment demonstrated reduced systemic exposure of SITA without compromising on its antihyperglycemic activity and improvement in conditions associated with diabetes.

Effects of alpha-tocopherol on acute pancreatitis in rat.

Acute pancreatitis is a disease with high morbidity and mortality, despite all the advances in technology. The overall mortality rate of acute pancreatitis is 10%, whereas the mortality rate in infected necrotizing pancreatitis is approximately 35%. In this study, we aimed to establish acute pancreatitis in rats in order to try out the alpha-tocopherol treatment protocol and to reveal the results biochemically and histopathologically. Twenty-four male male Sprague-Dawley rats weighing between 300 and 350 g were used in the study. In Group 1, 80 µg/kg of normal saline was subcutaneously injected into eight rats; in Group 2, 80 µg/kg of cerulein was subcutaneously injected into eight rats; and in Group 3, 80 µg/kg of cerulein was subcutaneously injected into eight rats. In addition, 30 mg/kg of alpha-tocopherol was intraperitoneally injected into eight rats. The mean Schoenberg score, serum amylase, and lipase and Neutrophil Gelatinase-Associated Lipocalin (NGAL) levels were statistically significantly higher in Group 2 than in Group 1. The mean Schoenberg score and serum amylase and lipase levels were statistically significantly lower in Group 3 than in Group 2. In this experimental study rat model of cerulein-induced acute pancreatitis, 30 mg/kg of alpha-tocopherol was injected intraperitoneally to examine its effect on pancreatitis. The improvement was observed in the histopathological examination of pancreatic tissues. We think that alpha-tocopherol may have a therapeutic effect on pancreatic tissue.

Quercetin improves endothelial function in diabetic rats through inhibition of endoplasmic reticulum stress-mediated oxidative stress

Endoplasmic reticulum (ER) stress attributes a crucial role in diabetes-induced endothelial dysfunction. The present study investigated the effects of quercetin, a potent antioxidant on the attenuation of ER stress-modulated endothelial dysfunction in streptozotocin (STZ)-induced diabetic rats. Oral administration of quercetin for six weeks to diabetic rats dose-dependently reduced the blood glucose levels and improved insulin secretion. Histopathological examination of pancreatic tissues in diabetic rats showed pathological changes such as shrunken islets, reduction in islet area and distorted β-cells, which were found to be restored by quercetin treatment. In addition, quercetin reduced the pancreatic ER stress-induced endothelial dysfunction as assessed by immunohistochemical analysis of C/ERB homologous protein (CHOP) and endothelin-1 (ET-1). Moreover, quercetin administration progressively increased the expression of vascular endothelial growth factor (VEGF) and its receptor, VEGFR2 in diabetes rats. Quercetin-mediated decrease in the nitric oxide (NO∙) and cyclic 3′,5′- guanosine monophosphate (cGMP) levels were also observed in the diabetic rats. Quercetin treatment reduced the lipid peroxidation in the diabetic rats, meanwhile increased the total antioxidant capacity in the pancreas from diabetic rats. Altogether, these results demonstrated the vasoprotective effect of quercetin against STZ-induced ER stress in the pancreas of diabetic rats.

Effects of Erdosteine on Experimental Acute Pancreatitis Model.

To create acute pancreatitis condition experimentally in rats using cerulein, and to reveal histopathological effects in pancreatic tissue with erdosteine. An experimental study. Department of General Surgery, Duzce University, Turkey, from June to October 2014. Thirty male Wistar albino rats were divided into three groups. No procedures were applied to Group 1. The rats in Group 2 and Group 3 were injected cerulein, to establish an experimental pancreatitis model and the blood amylase and lipase values were examined. The rats in Group 3 were given 10 mg/kg erdosteine. This treatment was continued for another 2 days and the rats were sacrificed. The pancreatic tissues were examined histopathologically for edema, inflammation, acinar necrosis, fat necrosis, and vacuolization. The lipase and amylase values and the histopathological examination of pancreatic tissues evidenced that the experimental acute pancreatitis model was established and edema, inflammation, acinar necrosis, fat necrosis, and vacuolization were observed in the pancreatic tissues. The statistical results suggest that erdosteine can decrease the edema, inflammation, acinar necrosis, fat necrosis and vacuolization scores in the tissues. The severity of acute pancreatitis, induced by cerulein in rats, is reduced with the use of erdosteine.

Ameliorative Effect of Adjunct Therapy of Metformin with Atorvastatin on Streptozotocin-induced Diabetes Mellitus in Rats.

Metformin has been used for the treatment of diabetes, whereas atorvastatin reduces the incidence of atherosclerosis and ischemic heart disease. Therefore, combined treatment with meformin plus atorvastatin may be beneficial in diabetic patients associated with cardiac disease. The present study was designed to evaluate the combination therapy of metformin and atorvastatin on streptozotocin-induced diabetes mellitus in rats. Blood pressure, serum insulin, glucose, lipid profiles and antioxidant enzymes in pancreatic tissues were measured. Histopathological examination of pancreatic tissues was performed. Streptozotocin treated rats showed significant decrease in body weight and body mass index. Streptozotocin-treated rats showed a significant increase in the levels of blood pressure, serum glucose, triglycerides, total cholesterol and thiobarbituric acid reactive substance as well as a significant decrease in the levels of serum insulin, high density lipoprotein and reduced glutathione in pancreatic tissues. Administration of metformin plus atorvastatin for a period of 14 days significantly improved these biochemical parameters near to normal. The protective effect of metformin plus atorvaststin against streptozotocin-induced diabetes was further confirmed by histopathological examination. The results of present study suggest that metformin plus atorvastatin possess antioxidant activity and has a significant protective effect against streptozotocin-induced diabetes mellitus.

Overexpression of spermidine/spermine N 1-acetyltransferase or treatment with N 1-N 11-diethylnorspermine attenuates the severity of zinc-induced pancreatitis in mouse

Depletion of pancreatic intracellular polyamine pools has been observed in acute pancreatitis both in the animal models and in humans. In this study, the wild-type mice, polyamine catabolic enzyme spermidine/spermine N(1)-acetyltransferase overexpressing (SSAT mice) and SSAT-deficient mice were used to characterize the new zinc-induced acute pancreatitis mouse model and study the role of polyamines and polyamine catabolism in this model. Intraperitoneal zinc injection induced acute necrotizing pancreatitis in wild-type mice as well as in SSAT-overexpressing and SSAT-deficient mice. Serum α-amylase activity was significantly increased in all zinc-treated mice compared with the untreated controls. However, the α-amylase activities in SSAT mice were constantly lower than those in the other groups. Histopathological examination of pancreatic tissue revealed edema, acinar cell necrosis and necrotizing inflammation, typical for acute pancreatitis. Compared with the other zinc-treated mice less damage according to the histopathological analysis was observed in the pancreatic tissue of SSAT mice. Levels of intracellular spermidine, and occasionally spermine, were significantly decreased in pancreases of all zinc-treated animals and SSAT enzyme activity was enhanced both in wild-type and SSAT mice. Interestingly, a spermine analog, N(1), N(11)-diethylnorspermine (DENSpm), enhanced the proliferation of pancreatic cells and reduced the severity of zinc-induced pancreatitis in wild-type mice. The results show that in mice a single intraperitoneal zinc injection causes acute necrotizing pancreatitis accompanied by decrease of intracellular polyamine pools. The study supports the important role of polyamines for the integrity and function of the pancreas. In addition, the study suggests that whole body overexpression of SSAT obtained in SSAT mice reduces inflammatory pancreatic cell injury.

Antidiabetogenic action of Morus rubra L. leaf extract in streptozotocin-induced diabetic rats.

Researchers all over the world are exploring herbal supplements to control diabetes and its complications. This study evaluated the antidiabetic action of Morus rubra L. aqueous leaf extract through its effect on hyperglycaemia, dyslipidaemia and oxidative stress in streptozotocin-induced diabetic rats. The extract was orally administered to diabetic rats (100, 200 and 400 mg/kg body weight) daily for 21 days. Fasting blood glucose was measured on days 0, 7, 14 and 21. At the end of the experiment, blood samples were drawn to measure glucose tolerance, glycosylated haemoglobin, insulin, C-peptide and lipid parameters. Antioxidant enzymes (superoxide dismutase and catalase), reduced glutathione and lipid peroxides were determined in blood and liver tissue. Histopathological examination of pancreatic tissue was also performed. The extract showed a dose-dependent fall in fasting blood glucose. Treatment with 400 mg/kg extract produced a significant reduction in glycosylated haemoglobin with a concomitant elevation in plasma insulin and C-peptide levels. The altered serum lipids in diabetic rats were significantly restored following treatment with the extract. In erythrocytes, as well as liver, the activity of antioxidant enzymes and content of reduced glutathione were found to be significantly enhanced, while levels of serum and hepatic lipid peroxides were suppressed in extract-fed diabetic rats. Histopathological examination of pancreatic tissue revealed an increased number of islets and beta-cells in extract-treated diabetic rats. M. rubra aqueous leaf extract leads to control over hyperglycaemia and dyslipidaemia. The study also demonstrates its antioxidant nature, and hence it may be protective against diabetic complications.

Induction of pancreatic acinar pathology via inhalation of nicotine.

This study was conducted to determine the effects of nicotine inhalation on the onset, progression, and sequential development of pancreatic lesions. Male Sprague-Dawley rats in groups of five were exposed to saline or nicotine aerosol twice daily for 15, 30, 45, and 60 min for 21 days. After sacrifice, blood samples were analyzed for plasma levels of nicotine, glucose, gastrin, and cholecystokinin. Pancreatic tissues were examined for pathological lesions. While there were no significant differences in plasma levels of glucose, gastrin, and cholecystokinin in all groups, there was a steady increase in plasma levels of nicotine with increased exposures to nicotine. Histopathological examination of pancreatic tissue revealed definitive pancreatic injuries that also appeared to be directly correlated with increased duration of nicotine exposure. The pathological changes of the pancreas were confined only to acinar cells of the exocrine pancreas. Two main types of cellular changes were observed: cellular swelling/vacuolation and nuclear condensation/cellular pyknosis. Both of these changes indicated tissue injuries in the pancreas. Transformation of the glandular acini to solid masses of epithelial cells was also observed. The results from our present study strongly suggest that the exocrine pancreas is very sensitive and susceptible to nicotine toxicity. Our data further indicate that early morphological changes in the pancreas induced by nicotine may occur without functional or metabolic alterations; however, such changes could occur at a later stage, when tissue and cellular changes become more extensive.

Hyperplasia and tumors of the islets of Langerhans in mice bearing an elastase I-SV40) T-antigen fusion gene

Transgenic mice bearing a fusion gene consisting of rat elastase I 5' flanking DNA fused to the early-coding (T-antigen) region of the SV40 genome (ELSV mice) develop carcinomas of the acinar cells of the exocrine pancreas. Histopathological examination of pancreatic tissue from 79 such animals revealed that, in addition to acinar cell neoplasms, ELSV mice develop two distinct lesions of the islets of Langerhans. By the age of 26 weeks, 36% of the mice had developed insulinomas. Starting at 8 weeks of age, the mice also developed D (somatostatin)-cell hyperplasia, which began at the periphery of the islets but which in advanced cases resulted in nearly complete replacement of other islet cell types by D-cells. By 26 weeks of age, 85% of the mice examined demonstrated the D-cell abnormality. Both the insulinomas and D-cell lesions were negative by immunohistochemistry for T-antigen, which was, however, demonstrated in acinar cell neoplasms using a monoclonal antibody against a C-terminal T-antigen epitope. Insulinomas have been described in other SV40 transgenic mice, particularly when the SV40 enhancer is not included in the transgene, suggesting that the presence of native SV40 enhancer may ordinarily suppress the expression of T-antigen in pancreatic beta-cells. The somatostatin-cell lesion is unique to the ELSV model; it may be neoplastic or represent a response to the growth and neoplastic changes occurring simultaneously in the exocrine pancreas.