Pieces Of Pancreas Research Articles

IDENTIFICATION OF EURYTREMA SPP. IN BRAHMAN CROSS CATTLE SLAUGHTERED IN BANDA ACEH SLAUGHTERHOUSE

Eurytrema spp. is a trematode worm that generally attacks the pancreas of livestock and humans which can cause chronic interstitial pancreatitis, weight loss that leads to decreased livestock productivity and ends in death. The purpose of this study is to identify the parasitic worm Eurytrema spp. and also to find out the prevalence rate in Brahman Cross cattle slaughtered in Banda Aceh slaughterhouse. This study used a sample of 20 pieces of Brahman Cross cattle pancreas from the Banda Aceh slaughterhouse. The worms were collected by taking the pancreas of Brahman Cross cattle that had been cut and stained with the Semicon Carmine staining method. The parameters of this study were the morphological differences in size of the body, oral sucker, ventral sucker and reproductive organs in each species of Eurytrema spp. The results showed that out of the 10 worms found, there was only 1 species, namely E. pancreaticum, which has the characteristics of a wide body with a length of 8-16 mm, the oral sucker is larger than the ventral sucker and lobed testes and ovaries. The prevalence rate of infection with E. pancreaticum is 15%. It was concluded that the worm that infects the Brahman Cross cattle slaughtered in Banda Aceh slaughterhouse is E. pancreaticum.

Abstract IA11: Three-dimensional genomic mapping of human pancreatic tissue reveals striking multifocality and genetic heterogeneity in precancerous lesions

Abstract Pancreatic intraepithelial neoplasia (PanIN) is a precursor to pancreatic cancer and represents a critical opportunity for cancer interception. However, the number, size, shape, and connectivity of PanINs in human pancreatic tissue samples are largely unknown. In this study, we quantitatively assessed human PanINs using CODA, a novel machine-learning pipeline for 3D image analysis that generates quantifiable models of large pieces of human pancreas with single-cell resolution. Using a cohort of 38 large slabs of grossly normal human pancreas from surgical resection specimens, we identified striking multifocality of PanINs, with a mean burden of 13 spatially separate PanINs per cm3 of sampled tissue. Extrapolating this burden to the entire pancreas suggested a median of approximately 1000 PanINs in an entire pancreas. In order to better understand the clonal relationships within and between PanINs, we developed a pipeline for CODA-guided multi-region genomic analysis of PanINs, including targeted and whole exome sequencing. Multi-region assessment of 37 PanINs from eight additional human pancreatic tissue slabs revealed that almost all PanINs contained hotspot mutations in the oncogene KRAS, but no gene other than KRAS was altered in more than 20% of the analyzed PanINs. PanINs contained a mean of 13 somatic mutations per region when analyzed by whole exome sequencing. The majority of analyzed PanINs originated from independent clonal events, with distinct somatic mutation profiles between PanINs in the same tissue slab. A subset of the analyzed PanINs contained multiple KRAS mutations, suggesting a polyclonal origin even in PanINs that are contiguous by rigorous 3D assessment. This study leverages a novel 3D genomic mapping approach to describe, for the first time, the spatial and genetic multifocality of human PanINs, providing important insights into the initiation and progression of pancreatic neoplasia. Citation Format: Laura D. Wood. Three-dimensional genomic mapping of human pancreatic tissue reveals striking multifocality and genetic heterogeneity in precancerous lesions [abstract]. In: Proceedings of the AACR Special Conference: Targeting RAS; 2023 Mar 5-8; Philadelphia, PA. Philadelphia (PA): AACR; Mol Cancer Res 2023;21(5_Suppl):Abstract nr IA11.

OR22-4 Homing of Antigen-specific Engineered Regulatory T Cells To Human Pancreatic Islets

Abstract The therapeutic application of regulatory T cells (Tregs) for the treatment of autoimmune disorders, although efficacious, has been limited by the scarcity of antigen-specific Tregs. If antigen-specific Tregs, capable of reaching the desired autoimmune target, could be produced on demand, antigen-specific immune suppression of autoimmune diseases would be achievable. One approach to endow T cells with a desired antigen-specificity uses chimeric T cell antigen-receptors (CAR) with antibody-type specificity. Adoptive cell transfer therapies with CAR-re-directed cytotoxic T cells, have shown impressive efficacy in the treatment of hematologic malignancies. Accordingly, employing such technology to re-direct Tregs to sites of autoimmune attack may be a useful therapeutic approach to alleviate a broad spectrum of diseases in which uncontrolled auto and alloimmune responses play a major role. We recently developed pancreatic beta-cell, antigen-specific, CAR Tregs (1) and explored their therapeutic potential against T1D in our humanized mouse model (2). Ours is the first successful, antigen-specific CAR-Treg treatment of T1D in a humanized mouse model that closely resembles the human disease. Based on our mice data, we believe treatment with pancreatic beta-cell, antigen-specific CAR-Tregs will allow for recovery and reconstitution of beta cells in human T1D patients as well. The purpose of this study was to determine if antigen-specific human CAR Tregs could also identify target and home to human pancreatic islets in culture as they do in mice. The study involved drawing 10 cc of blood 1-2 weeks prior to pancreas surgery; followed by collection of a small piece of pancreas (5 cc wedge) once the pancreas was removed for a clinical indication (cancer, pancreatitis). We first isolated Tregs from peripheral blood of these human donors and expanded them in vitro. Treg cells were genetically modified to express either a beta-cell antigen-specific (GAD65) CAR or an irrelevant (EPCAM) CAR construct together with GFP marker. CAR Tregs were selectively expanded in the presence of rhGAD65 antigen. Once pancreas tissue became available, it was processed for islet separation using the collagenase method. Pancreatic islets were then co-cultured with syngeneic CAR Tregs for 7 days. IncuCyte S3 Live-cell System (Sartorius) is a real-time system that automatically acquires and analyzes HD, phase and fluorescent images of cell cultures, around the clock, for 7 days, while cells remain undisturbed. Live immunofluorescence microscopy demonstrated the distinct homing of GAD65 CAR Tregs to islets as compared to control EPCAM CAR Tregs as early as 24 hours of co-culture. Importantly, proliferation of GAD65 CAR Tregs was clearly demonstrated within 72 hours. GAD65 CAR Treg cytokine profile from co-culture supernatant was characteristic of activated Tregs. 1 PATENT #WO2020097546 2 PATENT #US20200037586 Presentation: Monday, June 13, 2022 11:45 a.m. - 12:00 p.m.

Abdominal pain, diabetes mellitus and an absent piece of pancreas

Abdominal pain, diabetes mellitus and an absent piece of pancreas

Islet Cell Transplantation for Patients with Chronic Pancreatitis.

The pancreas is a dual organ. The bulk (98 %) consists of the exocrine acinar tissue and the islets of Langerhans, responsible for the endocrine component constituting only about 2 % of the organ. Nevertheless, loss of this endocrine component results in severe type 3c diabetes. The blood sugar control in these patients is very labile, perhaps because of the absence of both insulin and glucagon from loss of both beta and alpha cells of the islet of Langerhans. Chronic pancreatitis typically affects the exocrine component of the gland, and the islet structure and function is preserved unless the disease is in late advanced stages. Therefore, if total or partial pancreatectomy is indicated for pain control in chronic pancreatitis, islets can beseparated from the resectedpancreas, and thentransplanted back into the patient, to prevent development of diabetes [1]. Paul Langerhans described the eponymous islets in the pancreas in 1869, before their function was even known. Minkowsky in 1892 first reported the development of diabetes, in dogs, after removal of the pancreas [2]. He also described that the subcutaneous placement of a portion of the pancreas prevented the mortality associatedwith the total pancreatectomy. Watson Williams in 1893 [2] attempted to treat diabetes, in a 15-year-old boy, by subcutaneous placement of pieces of sheep pancreas. The patient survived for a few days only, but perhaps can be considered a first attempt at a pancreatic xenograft. With the discovery of insulin from purified

Improved Protocol For Laser Microdissection Of Human Pancreatic Islets From Surgical Specimens

Laser microdissection (LMD) is a technique that allows the recovery of selected cells and tissues from minute amounts of parenchyma 1,2. The dissected cells can be used for a variety of investigations, such as transcriptomic or proteomic studies, DNA assessment or chromosomal analysis 2,3. An especially challenging application of LMD is transcriptome analysis, which, due to the lability of RNA 4, can be particularly prominent when cells are dissected from tissues that are rich of RNases, such as the pancreas. A microdissection protocol that enables fast identification and collection of target cells is essential in this setting in order to shorten the tissue handling time and, consequently, to ensure RNA preservation. Here we describe a protocol for acquiring human pancreatic beta cells from surgical specimens to be used for transcriptomic studies 5. Small pieces of pancreas of about 0.5-1 cm3 were cut from the healthy appearing margins of resected pancreas specimens, embedded in Tissue-Tek O.C.T. Compound, immediately frozen in chilled 2-Methylbutane, and stored at -80 °C until sectioning. Forty serial sections of 10 μm thickness were cut on a cryostat under a -20 °C setting, transferred individually to glass slides, dried inside the cryostat for 1-2 min, and stored at -80 °C. Immediately before the laser microdissection procedure, sections were fixed in ice cold, freshly prepared 70% ethanol for 30 sec, washed by 5-6 dips in ice cold DEPC-treated water, and dehydrated by two one-minute incubations in ice cold 100% ethanol followed by xylene (which is used for tissue dehydration) for 4 min; tissue sections were then air-dried afterwards for 3-5 min. Importantly, all steps, except the incubation in xylene, were performed using ice-cold reagents - a modification over a previously described protocol 6. utilization of ice cold reagents resulted in a pronounced increase of the intrinsic autofluorescence of beta cells, and facilitated their recognition. For microdissection, four sections were dehydrated each time: two were placed into a foil-wrapped 50 ml tube, to protect the tissue from moisture and bleaching; the remaining two were immediately microdissected. This procedure was performed using a PALM MicroBeam instrument (Zeiss) employing the Auto Laser Pressure Catapulting (AutoLPC) mode. The completion of beta cell/islet dissection from four cryosections required no longer than 40-60 min. Cells were collected into one AdhesiveCap and lysed with 10 μl lysis buffer. Each single RNA specimen for transcriptomic analysis was obtained by combining 10 cell microdissected samples, followed by RNA extraction using the Pico Pure RNA Isolation Kit (Arcturus). This protocol improves the intrinsic autofluorescence of human beta cells, thus facilitating their rapid and accurate recognition and collection. Further improvement of this procedure could enable the dissection of phenotypically different beta cells, with possible implications for better understanding the changes associated with type 2 diabetes.

Improved Protocol For Laser Microdissection Of Human Pancreatic Islets From Surgical Specimens

Laser microdissection (LMD) is a technique that allows the recovery of selected cells and tissues from minute amounts of parenchyma. The dissected cells can be used for a variety of investigations, such as transcriptomic or proteomic studies, DNA assessment or chromosomal analysis. An especially challenging application of LMD is transcriptome analysis, which, due to the lability of RNA, can be particularly prominent when cells are dissected from tissues that are rich of RNases, such as the pancreas. A microdissection protocol that enables fast identification and collection of target cells is essential in this setting in order to shorten the tissue handling time and, consequently, to ensure RNA preservation. Here we describe a protocol for acquiring human pancreatic beta cells from surgical specimens to be used for transcriptomic studies. Small pieces of pancreas of about 0.5-1 cm(3) were cut from the healthy appearing margins of resected pancreas specimens, embedded in Tissue-Tek O.C.T. Compound, immediately frozen in chilled 2-Methylbutane, and stored at -80 °C until sectioning. Forty serial sections of 10 μm thickness were cut on a cryostat under a -20 °C setting, transferred individually to glass slides, dried inside the cryostat for 1-2 min, and stored at -80 °C. Immediately before the laser microdissection procedure, sections were fixed in ice cold, freshly prepared 70% ethanol for 30 sec, washed by 5-6 dips in ice cold DEPC-treated water, and dehydrated by two one-minute incubations in ice cold 100% ethanol followed by xylene (which is used for tissue dehydration) for 4 min; tissue sections were then air-dried afterwards for 3-5 min. Importantly, all steps, except the incubation in xylene, were performed using ice-cold reagents - a modification over a previously described protocol. utilization of ice cold reagents resulted in a pronounced increase of the intrinsic autofluorescence of beta cells, and facilitated their recognition. For microdissection, four sections were dehydrated each time: two were placed into a foil-wrapped 50 ml tube, to protect the tissue from moisture and bleaching; the remaining two were immediately microdissected. This procedure was performed using a PALM MicroBeam instrument (Zeiss) employing the Auto Laser Pressure Catapulting (AutoLPC) mode. The completion of beta cell/islet dissection from four cryosections required no longer than 40-60 min. Cells were collected into one AdhesiveCap and lysed with 10 μl lysis buffer. Each single RNA specimen for transcriptomic analysis was obtained by combining 10 cell microdissected samples, followed by RNA extraction using the Pico Pure RNA Isolation Kit (Arcturus). This protocol improves the intrinsic autofluorescence of human beta cells, thus facilitating their rapid and accurate recognition and collection. Further improvement of this procedure could enable the dissection of phenotypically different beta cells, with possible implications for better understanding the changes associated with type 2 diabetes.

A missing piece of pancreas

A 35-year-old man came for a second opinion because of a hepatic lesion noted by ultrasound at a local clinic. The physical examination was unremarkable. Abdominal ultrasound showed a 1.2 cm subcapsular hypoechoic area in segment VI of the liver on a background of increased echogenicity, which was confirmed to be focal fatty sparing on subsequent computed tomography (CT) study of the abdomen. CT imaging disclosed that the tail and a portion of the body of the pancreas were absent (Figure 1), while …

Pancreatic Injury Response is Different Depending on the Method of Resecting the Parenchyma

The present study was performed to compare the pancreatic injury response on the parenchymal resection either with ultrasonic scissors, electrocautery, or surgical scalpel. A 1 x 0.5 cm piece of rat pancreas was resected from side of the pancreas either with ultrasonic scissors (Harmonic Scalpel; UltraCision, Ethicon Endosurgery Inc., Cincinnati, OH) or electrocautery (Force FX; Valleylab, Tyco Healthcare Group LP, Boulder, CO) at two power levels, 1 and 3; 8W and 25W, respectively, or with surgical scalpel. Hemostasis was provided after surgical scalpel either with cellulose patch (Interceed; Johnson and Johnson Medical, Inc., New Brunswick, NJ), three stitches of 6-0 polydioxanone at tightness of 0.6N or fibrin glue (Tisseel Duo Quick; Baxter AG, Wien, Austria). Blood sample and pancreas specimens, both at the resection site and far away, were taken 1, 7, and 21 days postoperatively from exposed animals, sham operated animals (n = 18 in each) and from unexposed baseline animals (n = 5). Necrosis, edema, leukocyte infiltration, hemorrhage, vacuolization, and fibrosis were histologically assessed separately. Each resection and sham operation induced similar increase in the amylase activity on day 1 with normalization by day 7. Resection with ultrasonic scissors and electrocautery induced more tissue injury to the pancreas than resection with surgical scalpel independent of the method for hemostasis. The injury, although somewhat milder in intensity, was also observed in parts of the pancreas located far away from the site of resection. Of the compared methods, surgical scalpel resection plus cellulose patch or fibrin glue hemostasis induced the least histological changes in the pancreatic parenchyma. This injury response spread over the pancreas.

A modified automated method for isolation of viable pancreatic islets in laboratory animals

To achieve successful islet transplantation, a high viability is required. For this reason an automated method including two chambers: one for islets isolation and one for recirculation and collection was developed. Recently, we produced a modified version of this work by building a similar system of glass where marbles were not used for disaggregation, and the pancreatic tissue had to be disrupted mechanically before the digestion phase. By using the reconfigured system, we obtained 260 ± 20 islets from each Wistar albino rat (weighing 220 to 240 g) pancreas. Islets were observed at 35 minutes after the start of perfusion (closed circuit) and the optimum time to stop the isolation determined to be 40 minutes based upon islets viability. Although the present system is configured for islet isolation from small laboratory animals (rat, mouse), we have also obtained thousands of islets at 25 minutes after treatment of a 0.5-g piece of pig pancreas. Compared to the time-consuming manual method usually used for islet isolation from small laboratory animals, the new technique is economic, easy to use, and does not reduce islets viability.

Oncogene expression on the syngeneic β-cells of long-term surviving pancreatic grafts and better effects of interleukin-1 receptor (IL-1R) and IL-2Rβ on the grafted β-cells in LEW/Sea strain rats

Thirty-two normal LEW/Sea rats were transplanted a piece of syngeneic pancreas between the peritoneum and abdominal muscle. Among them, 17 (68%) of the 25 rats that received pancreatic transplantation at 41–50 days of age had a surviving β-cell mass at 5.5–7.1 months after transplantation. Among the 25 rats, 12 rats injected with interleukin-1 receptor (IL-1R) and IL-2Rβ peptides at post-transplantation showed better surviving grafts at 5.5 months’ observation. Only 2 (25%) of the other 7 young rats that received a pancreatic graft at 20 days of age had a small mass at 21 days post-transplantation. Flow cytometer (FCM) analyses showed that thymus OX40 + (CD134 +) T-cells were increased up to 37±4% at the graft rejection in the 13 old rats without the IL-R peptide injections. The 7 young rats had 99% of thymus OX40 + T-cells. However, the 12 old rats injected with the IL-R peptides showed suppressed numbers of thymus OX40 + T-cells (8–13±3%). The long-term surviving, but apoptotic, grafted β-cells were stained positively both with anti-insulin monoclonal antibody (mAb) and with anti-c-erbB-2/human epidermal growth factor receptor (HER)-2/neu mAb. Expression of a c-erb family oncogene was shown on the pancreatic graft surviving for 7.1 months. Electron microscopic analysis of the grafted β-cells showed abnormally large β granules and loss of functioning mitochondria in the cytoplasm. In 18 (56%) of the 32 rats, the 220-bp and 380-bp specific products of insulin-degrading enzyme (IDE) gene were amplified using the polymerase chain reaction (PCR) of the liver DNA. Among the 18 rats, 6 rats expressed 2 extra hands of 280-bp and 700-bp in a correlation with the high levels of the transforming growth factor-alpha (TGF-α) cDNA of 120-bp which was amplified in the quantitative reverse-transcriptase (RT)-PCR of the liver cDNA. Among the selected 11 rats, 5 rats showed large amounts of the 120-bp TGF-α cDNA. Host pancreatic RT-PCR showed 235-bp or 250-bp bcl-2 and 181-bp bcl-xS gene products. The bcl-2 cDNA of the host pancreas was amplified actively when the pancreatic graft was being rejected. Exceptionally, the one female injected with the IL-R peptides showed a low level of the liver TGF-α cDNA together with the pancreatic expressions of Bax (140-bp), bcl-2 and like interleukin converting enzyme (LICE) (318-bp) cDNA. Insulin secretion from the grafted β-cells and IL-1β-induced Fas-mediated apoptosis of the β-cells were suspected to be present at the same time in the female with the best graft survival.

Pancreatic lymphatic system in rodents.

The lymphatic network of the pancreas has been little investigated and recent studies have provided contrasting data. This research is aimed to supply the morphologic basis to outline the involvement of the lymphatic system in pancreatic pathology. Guinea pigs, rats, and mice were anesthetized with ether and sacrificed with the same anesthetic. Pieces of pancreas were processed for transmission electron microscopy. Semithin sections were observed by light microscopy and, after positive identification by transmission electron microscopy, lymphatics were followed with long series of consecutive sections to define their distribution. Lymphatics were detected in the pancreas of all the animals both in the inter and the intralobular sites. Closer relations with the exocrine parenchyma (ducts and acini) were observed in guinea pig pancreas. Remarkably, interesting relationships between lymphatics and endocrine tissue were observed in all the animals. Overall, however, the lymphatic network of rat pancreas was less develop and preferentially associated with blood vessels. The distribution of the pancreatic lymphatic network appears consistent with an active role in pancreatic pathology.

Pancreatic uptake of [2-14C]alloxan

The uptake of [2-(14)C]alloxan by the pancreatic gland was investigated in control and streptozotocin-induced diabetic (STZ) rats, using both in vitro and in vivo techniques. Whether after 10 to 60 min incubation of pieces of pancreas in the presence of [2-(14)C]alloxan or 60 min to 24 h after intravenous injection of [2-(14)C]alloxan to control and insulin-treated STZ rats, the radioactive content of the pancreas (dpm/mg wet weight) only represented, in the STZ rats, about two thirds of the reference value found in control animals. These findings indicate that insulin-producing islet B-cells participate to a sizeable extent to the overall uptake of [2-(14)C]- alloxan by the whole pancreatic gland, despite the fact that they account for no more than about one percent of the total pancreas mass. Hence, it should be possible to preferentially label the endocrine moiety of the pancreas, in the perspective of its imaging and quantification by a non-invasive procedure, by use of a suitable radiolabelled molecule selectively taken up by islet, as distinct from acinar, pancreatic cells.

Pancreatic fate of 14C-labelled hexoses.

In order to assess the respective contribution of the exocrine and endocrine moieties of the pancreas to the overall net uptake of selected monosaccharides by the pancreatic gland, the apparent distribution space of L-[1-14C]glucose, 3-O-[14C-methyl]-D-glucose, D-[U-14C]glucose, D-[U-14C]mannose and D-[U-14C]fructose was measured in pieces of pancreas obtained from either control rats or animals injected with streptozotocin. Although the time course for the uptake of 3-O-[14C-methyl]-D-glucose, D-[U-14C]glucose, D-[U-14C]mannose and D-[U-14C]fructose was much slower in the pieces of pancreas than that previously documented in isolated pancreatic islets, no significant difference could, as a rule, be detected between the results obtained in pancreatic pieces of control and streptozotocin rats. A comparable situation prevailed in the pancreas of animals examined 3 min after the intravenous injection of 3-O-[14C-methyl]-D-glucose. D-Glucose inhibited the uptake of 3-O-[14C-methyl]-D-glucose and that of D-[U-14C]fructose. Likewise, 3-O-methyl-D-glucose inhibited the uptake of D-[U-14C]glucose. Cytochalasin B (20 microm) also inhibited the uptake of 3-O-[14C-methyl]-D-glucose and D-[U-14C]glucose, but not that of D-[U-14C]fructose. D-Mannoheptulose hexaacetate, but not the unesterified heptose, inhibited the metabolism of tritiated and 14C-labelled D-glucose, as well as the net uptake of D-[U-14C]glucose and D-[U-14C]mannose and, to a lesser extent, that of D-[U-14C]fructose. These findings indicate that despite marked differences between endocrine and exocrine pancreatic cells in terms of both the time course for the uptake of several hexoses and the inhibition of their phosphorylation by D-mannoheptulose, little or no preferential labelling of the endocrine moiety of the pancreas by the 14C-labelled hexoses is observed, at least when judged from their distribution space in pancreatic pieces or the whole pancreatic gland. Nevertheless, the findings made with D-mannoheptulose and its hexaacetate ester raise the view that this heptose could conceivably be used to achieve a sizeable preferential labelling of the endocrine pancreas under the present experimental conditions.

Pancreatic fate of14C-labelled hexoses

In order to assess the respective contribution of the exocrine and endocrine moieties of the pancreas to the overall net uptake of selected monosaccharides by the pancreatic gland, the apparent distribution space of L-[1-14C]glucose, 3-O-[14C-methyl]-D-glucose, D-[U-14C]glucose, D-[U-14C]mannose and D-[U-14C]fructose was measured in pieces of pancreas obtained from either control rats or animals injected with streptozotocin. Although the time course for the uptake of 3-O-[14C-methyl]-D-glucose, D-[U-14C]glucose, D-[U-14C]mannose and D-[U-14C]fructose was much slower in the pieces of pancreas than that previously documented in isolated pancreatic islets, no significant difference could, as a rule, be detected between the results obtained in pancreatic pieces of control and streptozotocin rats. A comparable situation prevailed in the pancreas of animals examined 3 min after the intravenous injection of 3-O-[14C-methyl]-D-glucose. D-Glucose inhibited the uptake of 3-O-[14C-methyl]-D-glucose and that of D-[U-14C]fructose. Likewise, 3-O-methyl-D-glucose inhibited the uptake of D-[U-14C]glucose. Cytochalasin B (20 microm) also inhibited the uptake of 3-O-[14C-methyl]-D-glucose and D-[U-14C]glucose, but not that of D-[U-14C]fructose. D-Mannoheptulose hexaacetate, but not the unesterified heptose, inhibited the metabolism of tritiated and 14C-labelled D-glucose, as well as the net uptake of D-[U-14C]glucose and D-[U-14C]mannose and, to a lesser extent, that of D-[U-14C]fructose. These findings indicate that despite marked differences between endocrine and exocrine pancreatic cells in terms of both the time course for the uptake of several hexoses and the inhibition of their phosphorylation by D-mannoheptulose, little or no preferential labelling of the endocrine moiety of the pancreas by the 14C-labelled hexoses is observed, at least when judged from their distribution space in pancreatic pieces or the whole pancreatic gland. Nevertheless, the findings made with D-mannoheptulose and its hexaacetate ester raise the view that this heptose could conceivably be used to achieve a sizeable preferential labelling of the endocrine pancreas under the present experimental conditions.

D-mannoheptulose uptake and its metabolic and secretory effects in human pancreatic islets.

D-mannoheptulose was recently proposed to be transported into cells at the intervention of GLUT2. Since GLUT1, rather than GLUT2, represents the major carrier system for the transport of monosaccharides across the islet B-cell plasma membrane in human subjects, the uptake of D-mannoheptulose and its metabolic and secretory effects were investigated in human islets. The uptake of D-glucose reached much more rapidly a close-to-equilibrium value in isolated islets than in pieces of pancreas obtained from the same donor. The distribution space of D-[3H]mannoheptulose in the human islets largely exceeded that of [U-14C]sucrose, considered as an extracellular marker, and did not differ significantly from that of 3HOH. In the human islets, the heptose (10.0 mM) inhibited both D-[5-3H]glucose utilization and D-[U-14C] glucose oxidation, and decreased glucose-stimulated insulin release to the same extent as D-mannoheptulose hexaacetate. These findings indicate that a suitable radioactive analog of D-mannoheptulose could be used, in human like in rat islets, for preferential labelling of the endocrine moiety of the pancreatic gland.

Possible Relationship between the B-Cell Threshold for Glucose-Induced Insulin Secretion and Blood Glucose Concentrations in the Normal Toad

The aim of this work was to gain information on the possible relationship between basal glycemia in the toad and the B-cell threshold for glucose-induced insulin release. Hence, pieces of pancreas from Bufo arenarum were incubated with 2 to 20 mM glucose or preincubated with 2 mM glucose plus the hexokinase and glucokinase inhibitors (50 mM of 2-deoxyglucose and mannoheptulose, respectively) followed by an incubation with different glucose concentrations. The maximal rate of insulin release occurred at 8 mM glucose, while 50% of the release (Ks50) was observed at 7 mM glucose. Regardless of the glucose concentration employed, pancreas pieces preincubated with 2-deoxyglucose released less insulin than the corresponding controls. On the other hand, mannoheptulose significantly inhibited the release of insulin at high glucose concentrations, having no effect at low glucose concentrations. The blocking effect of these two inhibitors is the first indirect evidence of the existence of the hexokinase/glucokinase enzymic system in the toad pancreas. Since the activity ratio of this system determines the glucose sensitivity of the insulin secretory mechanism, it is concluded that the possible existence of a higher ratio of these enzymes in toad B cells could explain the particular characteristics of glucose sensitivity in this animal, which in turn may explain its low blood glucose concentration.

Uptake of Tritiated Glibenclamide by Endocrine and Exocrine Pancreas

Tritiated glibenclamide binds to specific receptors and is internalized in pancreatic insulin-producing B-cells. We investigated, therefore, whether tritiated glibenclamide could be used to preferentially label the endocrine, as distinct from exocrine, pancreas. In isolated rat pancreatic islets, the net uptake of 3H-glibenclamide reached within 30 min of incubation a near-equilibrium value, corresponding to an apparent distribution space close to three to four times the islet volume. In pieces of pancreas exposed up to 1 h to 3H-glibenclamide, however, its apparent distribution space progressively increased and, even at the min 60 of incubation, did not exceed a third of the wet weight of the pieces. Yet, no significant difference could be detected between the time course for 3H-glibenclamide uptake by pancreatic pieces from either control animals or rats injected with streptozotocin a few days before the experiments. Likewise, no significant difference in the paired ratio between the radioactive content of the pancreas and plasma could be found between the control and diabetic rats when examined 1, 5, or 24 h after the IV administration of 3H-glibenclamide. These findings indicate that the sulfonylurea does not represent a suitable tool for preferential labeling of the endocrine pancreas in the perspective of its imaging by a noninvasive procedure.

Uptake of tritiated glibenclamide by endocrine and exocrine pancreas.

Tritiated glibenclamide binds to specific receptors and is internalized in pancreatic insulin-producing B-cells. We investigated, therefore, whether tritiated glibenclamide could be used to preferentially label the endocrine, as distinct from exocrine, pancreas. In isolated rat pancreatic islets, the net uptake of 3H-glibenclamide reached within 30 min of incubation a near-equilibrium value, corresponding to an apparent distribution space close to three to four times the islet volume. In pieces of pancreas exposed up to 1 h to 3H-glibenclamide, however, its apparent distribution space progressively increased and, even at the min 60 of incubation, did not exceed a third of the wet weight of the pieces. Yet, no significant difference could be detected between the time course for 3H-glibenclamide uptake by pancreatic pieces from either control animals or rats injected with streptozotocin a few days before the experiments. Likewise, no significant difference in the paired ratio between the radioactive content of the pancreas and plasma could be found between the control and diabetic rats when examined 1, 5, or 24 h after the IV administration of 3H-glibenclamide. These findings indicate that the sulfonylurea does not represent a suitable tool for preferential labeling of the endocrine pancreas in the perspective of its imaging by a noninvasive procedure.

Islet separation and islet cell culture in vitro from human embryo pancreas.

Diabetes mellitus is the most common disease and its death rate ranks the 8th in the world. Up to now, its incidence has a tendency to increase[1]. Since disorder of sugar metabolism might result in microvascular degeneration and injury of important human organs, it will endanger human health seriously. In 1969, Younszai first reported the method that could decrease diabetes symptoms by islet tissue transplantation. In 1981, our country began to treat diabetes type I by transplanting cultured islet tissues. In our study, we digested the pieces of pancreas with collagenase and made morphological observation on islet cells cultured for 3 d, 5 d, 7 d, 9 d and 13 d respectively, and the contents of insulin and C-peptide in the supernatant were detected by radioimmunoassay. The experimental model of rabbit diabetes was established, and certain curative effect was achieved in the treatment of experimental diabetes in rats.