Pancreatic Extracts Research Articles

Influence of Harvesting Stage on Phytochemical Composition, Antioxidant, and Antidiabetic Activity of Immature Ceratonia siliqua L. Pulp from Béni Mellal-Khénifra Region, Morocco: In Silico, In Vitro, and In Vivo Approaches.

Ceratonia siliqua L. is a medicinal plant that has long been used in traditional Moroccan medicine to treat many diseases. This study aimed to assess the impact of the stages of the immature phase of carob pulp (M1, M2, M3, M4, and M5) on phytochemical composition, antioxidant activity, and antidiabetic activity of Ceratonia siliqua L. The identification of the phenolic profile by HPLC-UV/MS-MS and the study of the antidiabetic effect by in silico, in vitro, and in vivo studies were carried out for extracts with high contents of phenolic compounds from immature wild carob pulp from the communes of Timoulit (TM), Bin Elouidane (AW), and Ouaouizerth (TG) in the province of Azilal in the Béni Mellal-Khénifra region. The results revealed a gradual increase in total sugar content over the pulp's ripening period, reaching a value of 2134 ± 56.23 mg GE/100 g fresh weight (FW) for TG. The three locations showed peak values for total polyphenol content (TPC), total flavonoid content (TFC), and total condensed tannin (TCT) at the M2 stage. AW had the highest concentrations of TPC (3819 ± 226.4 mg GAE/100 g FM), TFC (1034 ± 57.08 mg QE/100 g FM), and TCT (1472 ± 28.46 mg CE/100 g FM). The DPPH assay (7892 ± 296.1 mg TE/100 g FM) and the FRAP assay (278.2 ± 7.85 mg TE/100 g FM) both demonstrated that the TG zone is a highly potent antioxidant zone. In contrast, the AW site exhibited a markedly elevated value of 725.4 ± 103.6 mg TE/100 g FM in the ABTS assay. HPLC-UV-MS/MS analysis showed that the methanolic extracts of immature carob pulp (MEICP) from the three areas contained several different chemical compounds. The most prevalent were 3-O-p-coumaroyl-5-O-caffeoylquinic acid, quercetin 3-methyl ether, gallic acid, and galloylquinic acid. Immature carob pulp extract (ICPE) from AW showed the strongest in vitro inhibition of pancreatic α-amylase (IC50 = 0.405 µg/mL) and TG extracts were most potent against intestinal α-glucosidase (IC50 = 0.063 µg/mL). In vivo, AW, TG, and TM extracts significantly reduced postprandial glycemia in rats, with AW having the greatest effect. These results highlight the antidiabetic potential of ICPE. The 3-O-p-Coumaroyl-5-O-caffeoylquinic acid showed better affinity for α-amylase compared to acarbose and interacted significantly with several amino acid residues of the enzyme. Similarly, this molecule and 3,4-Dicaffeoylquinic acid demonstrated a strong affinity for α-glucosidase, suggesting their potential as natural inhibitors of enzymes involved in carbohydrate metabolism. Most of the compounds are not substrates of P-glycoprotein and exhibited high intestinal absorption. Furthermore, the majority of these compounds did not act as inhibitors or substrates of CYP450 enzymes, reinforcing their suitability for development as oral medications. These results underscore the potential of immature carob pulp as a promising antidiabetic agent.

The impact of using predigested feed on growth and molting frequency in blue swimming crab (Portunus pelagicus) enlargement

Abstract Blue swimming crab is one of the leading fishery export commodities which occupies the 3rd largest position after tuna and shrimp. Crab maintenance so far uses rasbora fish feed which has several drawbacks, including inappropriate nutrition, depending on the season, and is difficult to handle. Therefore, simple artificial feed is needed to support crab growth. The purpose of this study was to determine the effect of artificial feeding supplemented with pancreatic extract on crab growth. The research design used in this study was completely randomized design (CRD) with different concentrations of pancreatic extract (0%, 3%, 6%, 9%). Parameters observed included water quality (temperature, dissolved oxygen, pH, salinity, and ammonia), feed proximate, crab biology. The results of this study indicate that the water quality in this study supports the life of blue swimming crabs. The feed proximate results show that the proximate value is in accordance with the crab’s needs. The results of swimming crab biology showed that the highest growth of crab was owned by the artificial feed + 9% pancreas and the lowest was in the trash fish treatment. Based on the results it can be seen that the best treatment to support the life of the blue swimming crab is artificial feed + 9% pancreas. Predigestion feed with the help of pancreatic extract concentrations can increase crab growth compared to ordinary feed or rasbora fish feed.

Spatially resolved distribution of pancreatic hormones proteoforms by MALDI-imaging mass spectrometry

Zinc plays a crucial role both in the immune system and endocrine processes. Zinc restriction in the diet has been shown to lead to degeneration of the endocrine pancreas, resulting in hormonal imbalance within the β-cells. Proteostasismay vary depending on the stage of a pathophysiological process, which underscores the need for tools aimed at directly analyzing biological status. Among proteomics methods, MALDI-ToF-MS can serve as a rapid peptidomics tool for analyzing extracts or by histological imaging. Here we report the optimization of MALDI imaging mass spectrometry analysis of histological thin sections from mouse pancreas. This optimization enables the identification of the major islet peptide hormones as well as the major accumulated precursors and/or proteolytic products of peptide hormones. Cross-validation of the identified peptide hormones was performed by LC-ESI-MS from pancreatic islet extracts. Mice subjected to a zinc-restricted diet exhibited a relatively lower amount of peptide intermediates compared to the control group. These findings provide evidence for a complex modulation of proteostasis by micronutrients imbalance, a phenomenon directly accessed by MALDI-MSI.

Single-Cell Untargeted Lipidomics Using Liquid Chromatography and Data-Dependent Acquisition after Live Cell Selection.

We report the development and validation of an untargeted single-cell lipidomics method based on microflow chromatography coupled to a data-dependent mass spectrometry method for fragmentation-based identification of lipids. Given the absence of single-cell lipid standards, we show how the methodology should be optimized and validated using a dilute cell extract. The methodology is applied to dilute pancreatic cancer and macrophage cell extracts and standards to demonstrate the sensitivity requirements for confident assignment of lipids and classification of the cell type at the single-cell level. The method is then coupled to a system that can provide automated sampling of live, single cells into capillaries under microscope observation. This workflow retains the spatial information and morphology of cells during sampling and highlights the heterogeneity in lipid profiles observed at the single-cell level. The workflow is applied to show changes in single-cell lipid profiles as a response to oxidative stress, coinciding with expanded lipid droplets. This demonstrates that the workflow is sufficiently sensitive to observing changes in lipid profiles in response to a biological stimulus. Understanding how lipids vary in single cells will inform future research into a multitude of biological processes as lipids play important roles in structural, biophysical, energy storage, and signaling functions.

Obesity Model in White Male Wistar Rats: The Impact of Moringa (Moringa Oleifera) Flower Extract on Pancreatic Function as Deduced from Serum Amylase and Lipase Levels and the Histopathological Image of the Pancreas

Diabetes and obesity go hand in hand. Antidiabetic drugs are effective in treating diabetes. However, currently available antidiabetic drugs are too expensive, do not work, and cause major side effects. A significant therapeutic option for Diabetes Mellitus and obesity is using bioactive chemicals produced by plants. This compound is efficacious, easily accessible, safe, and affordable. The main aim of this study was to determine whether pancreatic extract from Moringa oleifera flowers affected blood amylase and lipase levels, as well as the histological appearance, of obese male Wistar white rats (Rattus norvegicus). This is a True experimental study, with the research design used as a test control Group Design. The research data was then analyzed with the help of SPSS using ANOVA and Post Hoc Test with LSD technique. This researcher used 24 male rats. Treatment with Moringa oleifera flower extract improves pancreatic function in obese Wistar white rats by reducing serum lipase and amylase levels. The results of research on serum lipase and amylase levels showed that a dose of 600mg/Kg BW improved pancreatic function in white rats of the Wistar strain that were obese.

In vitro gastrointestinal digestion of marine oil emulsions and liposomal solutions: fate of LC-PUFAs upon lipolysis.

The bioaccessibility and bioavailability of dietary fatty acids depend on the lipid to which they are esterified, the organisation of theses lipids in water and their recognition by lipolytic enzymes. In this work, we studied the release of marine long-chain polyunsaturated fatty acids (LC-PUFA), depending on their presentation either in the form of phospholipids (PL) or triacylglycerol (TAG). Two formulations based on marine PL or TAG extracted from salmon heads (Salmo salar) were prepared. Lipolysis was first tested in vitro by using individual gastrointestinal lipases and phospholipases to identify the enzymes involved in the digestion. Second, the lipolysis of the prepared formulations by a combination of enzymes was tested under in vitro conditions mimicking the physiological conditions found in the GI tract, both in the stomach and in the upper small intestine, in order to evaluate digestibility of TAG and LC-PUFA-containing liposomes. The in vitro results showed that TAG emulsion was hydrolyzed by porcine pancreatic extracts (PPE) and pure pancreatic lipase (PPL) with its cofactor, colipase, and to a lesser extent by pancreatic-lipase-related protein 2 (PLRP2) and a gastric extract (RGE) containing gastric lipase while no hydrolysis was observed with purified pancreatic phospholipase A2 (PLA2) and carboxyl ester hydrolase (CEH). The PL substrate was found to be hydrolysed by PLA2, PPE and PLRP2. Their phospholipase activities on liposomes formulation was dependent on the presence of bile salts. Using a two-step in vitro digestion model, we measured the kinetics of fatty acid release from TAG and PL during the gastric and intestinal phases of digestion. The highest overall lipolysis level was obtained with liposomes (around 75%) during the intestinal phase while they were preserved during the gastric phase. The overall lipolysis level of TAG emulsion was lower (around 33%), while it started already in the gastric phase. In conclusion, liposomes appear as a better delivery system for intestinal absorption of LC-PUFA than TAG. In addition, their resistance to lipolysis under gastric condition can protect LC-PUFA and provide a gastric stable delivery system for other molecules.

Lydia Marie DeWitt and Mary Butler Kirkbride: Prototypical Circa-Early-1900s Women of Pathology and an Analysis of Their Contributions to the Discovery of Insulin.

The year 2023 marks the centenary of the Nobel Prize honoring the discovery of insulin. Little-known experimental pathologists Lydia DeWitt, MD, at the University of Michigan and Mary Kirkbride, DSc [Hon], at Columbia University, both just beginning their academic careers, made independent contributions to the discovery that have never been critically examined. This happened at a time when it was exceedingly rare for women to work in pathology. To explore the facilitative roles of DeWitt and Kirkbride in the discovery of insulin and to examine their trail-breaking careers in academic pathology. Available primary and secondary historical resources were reviewed. DeWitt made and tested pancreatic extracts from duct-ligated atrophic pancreas (ie, Frederick Banting's great idea to prevent digestion of its hypothetical internal secretion) 15 years before Banting; Banting was unaware of her work. His idea came from reading a paper by pathologist Moses Barron. Prior duct-ligation studies had sometimes been viewed with skepticism because histologic identification of islets in atrophic duct-ligated pancreata was imperfect; Kirkbride addressed this with histochemical staining, convincing Barron and, therefore, indirectly influencing and motivating Banting. The lives and convoluted careers of these 2 early-20th-century women are explored and compared with those of other contemporary women in pathology. A unifying pattern becomes clear: careers in experimental pathology and bacteriology were accepted but performing clinical work in anatomic pathology was not. Both DeWitt and Kirkbride are prototypical early-20th-century women in academic pathology whose careers were constrained by gender. However, Kirkbride made a unique and unrecognized contribution to the discovery of insulin.

On the occasion of the centennial of the Nobel Prize in Physiology or Medicine, 1923: Nicolae C. Paulescu-between scientific creativity and political fanatism.

On the occasion of the centennial of the Nobel Prize in Physiology or Medicine, 1923: Nicolae C. Paulescu-between scientific creativity and political fanatism.

The Nobel Prize of Physiology or Medicine, 1923: controversies on the discovery of the antidiabetic hormone

AimsTo analyze the main contributions to the discovery of the antidiabetic hormone in the period between 1889, the year in which Oskar Minkowski demonstrated that complete pancreatectomy in dogs caused diabetes, and the year 1923, the date in which the clinical use of insulin was consolidated. A main objective has been to review the controversies that followed the Nobel Prize and to outline the role of the priority rule in Science.MethodsWe have considered the priority rule defined by Robert Merton in 1957, which takes into account the date of acceptance of the report of a discovery in an accredited scientific journal and/or the granting of a patent, complemented by the criteria set out by Ronald Vale and Anthony Hyman (2016) regarding the transfer of information to the scientific community and its validation by it. The awarding of the Nobel Prize in Physiology or Medicine in October 1923 has represented a frame of reference. The claims and disputes regarding the prioritization of the contributions of the main researchers in the organotherapy of diabetes have been analyzed through the study of their scientific production and the debate generated in academic institutions.Main results and conclusions(1) According to the criteria of Merton, Vale and Hyman, the priority of the discovery of the antidiabetic hormone corresponds to the investigations developed in Europe by E. Gley (1900), GL Zülzer (1908) and NC Paulescu (1920). (2) The active principle of the pancreatic extracts developed by Zülzer (acomatol), Paulescu (pancreina) and Banting and Best (insulin) was the same. (3) JB Collip succeeded in isolating the active ingredient from the pancreatic extract in January 1922, eliminating impurities to the point of enabling its use in the clinic. (4) In 1972, the Nobel Foundation modified the purpose of the 1923 Physiology or Medicine award to Banting and Macleod by introducing a new wording: "the credit for having produced the pancreatic hormone in a practical available form" (instead of “for the discovery of insulin”).

Glucagon and Insulin Overview: An Odd Couple's History and Physiology.

Glucagon is a peptide hormone that is produced primarily by the alpha cells in the islet of Langerhans in the pancreas, but also in intestinal enteroendocrine cells and in some neurons. Approximately 100 years ago several research groups discovered that pancreatic extracts would cause a brief rise blood glucose before they observed the decrease in glucose attributed to insulin. An overall description of the regulation of glucagon secretion requires inclusion of its sibling insulin because they both are made primarily by the islet and they both regulate each other in different ways. For example, glucagon stimulates insulin secretion, whereas insulin suppresses glucagon secretion. The mechanism of action of glucagon on insulin secretion has been identified as a trimeric guanine nucleotide-binding protein (G-protein)-mediated event. The manner in which insulin suppresses glucagon release from the alpha cell is thought to be highly dependent on the peri-portal circulation of the islet through which blood flows downstream from beta cells to alpha cells. In this scenario, it is via the circulation that insulin is thought to suppresses the release of glucagon. However, high levels of glucose also have been shown to suppress glucagon secretion. Consequently, the glucose lowering effect of insulin may be additive to the direct effects of insulin to suppress alpha cell function, so that in vivo both the discontinuation of the insulin signal and the condition of low glucose jointly are responsible for induction of glucagon secretion.

Safety and efficacy of a feed additive consisting of pancreatin from porcine pancreas (Pan-zoot) for dogs (Almapharm GmbH + Co KG).

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of a pancreatic extract (Pan-zoot) as a zootechnical additive for dogs. The EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) could not conclude on the safety of Pan-Zoot as a feed additive for dogs under the proposed conditions of use. The FEEDAP Panel could not conclude on the skin/eye irritancy potential of the additive or on the dermal sensitisation potential. Owing to its proteinaceous nature, the additive is considered a respiratory sensitiser. The additive may induce allergic reactions to the exposed users. The Panel concluded that there is no need for an environmental risk assessment. The FEEDAP Panel could not conclude on the efficacy of the product as a feed additive at the recommended conditions of use.

On the occasion of the centennial of insulin therapy (1922–2022), II-Organotherapy of diabetes mellitus (1906–1923): Acomatol. Pancreina. Insulin.

AimsThe general objective has been the historiographical investigation of the organotherapy of diabetes mellitus between 1906 and 1923 in its scientific, social and political dimensions, with special emphasis on the most relevant contributions of researchers and institutions and on the controversies generated on the priority of the "discovery" of antidiabetic hormone.MethodsWe have analyzed the experimental procedures and determination of biological parameters used by researchers during the investigated period (1906–1923): pancreatic ablation techniques, induction of acinar atrophy with preservation of pancreatic islets, preparation of pancreatic extracts (PE) with antidiabetic activity, clinical chemistry procedures (glycemia, glycosuria, ketonemia, ketonuria, etc.). The field investigation has included on-site and online visits to cities, towns, buildings, laboratories, universities, museums and research centers where the reported events took place, obtaining documents, photographic images, audiovisual recordings, as well as personal interviews complementary to the documentation consulted (primary sources, critical bibliography, reference works). The documentary archival sources have been classified according to theme, including those consulted in situ with those extracted online and digitized copies received mainly by email. Among the many archives contacted, those listed below have been most useful and have been consulted on site and on repeated visits: National Library of Medicine-Historical Archives (Bethesda, MD, USA); Archives, University of Toronto and Thomas Fisher Rare Books Library (Toronto, Ontario, Canada); Francis A. County Library of Medicine, Harvard University (Boston, Mass, USA); Zentralbibliothek der Humboldt-Universität (Berlin, DE), Geheimarchiv des Preuβischen Staates (Berlin, DE); Landesamt für Bürger—und Ordnungsangelegenheiten (LABO) (Berlin, DE); Arhivele Academiei Române şi Universitǎții Carol Davila (Bucharest, RO).Main results and conclusionsA) The European researchers Zülzer (Z Exp Path Ther 23:307–318, 1908) and Paulescu (CR Seances Soc Biol Fil 85:558, 1921) meet the requirements of the priority rule in the discovery of the antidiabetic hormone. B) Factors of socioeconomic and political nature related with the First World War and the inter-war period delayed the process of purification of the antidiabetic hormone in Europe. C) The Canadian scientist J. Collip, University of Alberta, temporarily assimilated to the University of Toronto, and the American chemist and researcher G. Walden, with the expert collaboration of Eli Lilly & Co., were the main authors of the purification process of the antidiabetic hormone. D) The scientific evidence, reflected in the heuristics of this research, allows to assert that the basic investigation carried out by the Department of Physiology of the University of Toronto, directed by the Scottish J. Macleod, in conjunction with the clinical research undertaken by the Department of Medicine of the University of Toronto (W. Campbell, A. Fletcher, D. Graham) made it possible in record time the successful treatment of patients with what was until then a deadly disease.

CFTR pharmacological modulators: A great advance in cystic fibrosis management

Cystic fibrosis is a severe monogenic disease that affects around 7400 patients in France. More than 2100 mutations in the cystic fibrosis conductance transmembrane regulator (CFTR), the gene encoding for an epithelial ion channel that normally transports chloride and bicarbonate, lead to mucus dehydration and impaired bronchial clearance. Systematic neonatal screening in France since 2002 has enabled early diagnosis of cystic fibrosis. Although highly demanding, supportive treatments including daily chest physiotherapy, inhaled aerosol therapy, frequent antibiotic courses, nutritional and pancreatic extracts have improved the prognosis. Median age at death is now beyond 30 years. Ivacaftor was the first CFTR modulator found to both reduce sweat chloride concentration and improve pulmonary function in the rare CFTR gating mutations. Combinations of modulators such as lumacaftor + ivacaftor or tezacaftor + ivacaftor were found to improve pulmonary function both in patients homozygous for the F508del mutation characterized by the lack of CFTR protein and those heterozygous for F508del with minimal CFTR activity. The triple combination of ivacaftor + tezacaftor + elexacaftor was recently shown to significantly improve pulmonary function and quality of life, to normalize sweat chloride concentration, and to reduce the need for antibiotic therapy in patients with at least one F508del mutation (83% in France). These impressive data, however, need to be confirmed in the long term. Nevertheless, it is encouraging to hear treated patients testify about their markedly improved quality of life and to observe that the number of lung transplants for cystic fibrosis decreased dramatically in France after 2020, despite the COVID pandemic, with no increase in deaths without lung transplant.

Pancreas Protective Effects of Telang Flower Extract (Clitoria ternatea L.) Against induced Male White Rats (Rattus norvegicus) Aloksan

Studies have been conducted on the protective effects of pancreatic telang flower extract (Clitoria ternatea L.) against white rats (Rattus norvegicus) induced by alloxane-induced by laboratory experiential methods. Thegoal is to determine the protective effect of pancreatic telang flower extract (Clitoria ternatea L.) against white rats (Rattus norvegicus) induced by alloxane which has the potential to be anti-diabetic. The mice used were 15 heads, the mice were divided into 5 groups. Each group consisted of 3 mice. Three groups were given a suspension of Telang Flower Extract at a dose of 150 mg/kgBB, 300 mg/kgBB, 450 mg/kgBB, and 1 group as a negative control was given a 1% Na-CMC suspension w/v. The administration of telang flower extract was carried out for 5 days and induced by aloksan of 150 mg / KgBB for 2 days. Blood draws are carried out 3 times, namely on the first day, day 6, and day 8 and then measurements of glucose and albumin levels are carried out. Based on the research that has been carried out, it can be concluded that the administration of telang flower extract (Clitoria ternatea L.) at a dose of 150 mg/KgBB,300 mg/KgBB, and 450 mg / KgBB have a protective effect not significantly different.

I-European research, the cradle of the discovery of the antidiabetic hormone: the pioneer roles and the relevance of Oskar Minkowski and Eugène Gley

AimsThe introduction of hormonal treatment in severe diabetes in 1922 represented a clinical and social impact similar to that of antibiotic therapy. In October 1923, the Assembly of the Karolinska Institute decided to award the Nobel Prize in Physiology or Medicine to the Canadian Frederick Grant Banting and the Scottish John James Rickard Macleod, researchers at the University of Toronto (UT), for "the discovery of insulin a year before". A few weeks later, European and American researchers protested the decision. The controversy remains to this day.MethodsWe have conducted a comprehensive review of primary and critical sources focused on theorganotherapy of animal and human diabetes mellitus since 1889, when Oskar Minkowski demonstrated the induction of experimental diabetes by total pancreatectomy in the dog, until the spring of 1923, when the Nobel Foundation had already received all the nominations for the award in Physiology or Medicine.ResultsThe in-depth analysis of all these sources revealed that Europe was the cradle of the discovery of the antidiabetic hormone. The discovery involved multiple research steps headed by a long list of key investigators, mainly European.ConclusionMarcel Eugène Émile Gley was the first to demonstrate the presence of the “antidiabetic principle” in extracts from “sclerosed” pancreas. The French physiologist pioneered the successful reduction of glycosuria and diabetic symptoms by the parenteral administration of pancreatic extracts to depancreatized dogs in experiments developed between 1890 and 1905, antedating insulin in two decades.

“It Works! Now What?” Insulin Development, Production, and Distribution at Connaught Laboratories, University of Toronto, 1922–24

The first successful human use of a pancreatic extract on Leonard Thompson, a 13-year-old severe diabetic at Toronto General Hospital, on 23 January 1922, is generally seen as the climax of the discovery-of-insulin story. However, this seminal event in the history of medicine begs the question: “It works! Now what?” The story may well have ended abruptly if not for the foresight and leadership of Dr. John G. FitzGerald, director of the University of Toronto’s Connaught Antitoxin Laboratories, based in the basement of the same Medical Building where F.G. Banting and C.H. Best, under the direction of J.J.R. Macleod, conducted the research that led to the first tests on Thompson. Significantly, it was the addition to the team of biochemist J.B. Collip and his purification of the extract that enabled the successful 23 January test. Within a day, however, tensions within the group, especially between Banting and Collip, and Collip’s threat to leave and take out his own patent, prompted FitzGerald to act. On 25 January, FitzGerald finalized a seminal agreement between Connaught and Banting, Best, Collip, and Macleod that secured the further development of methods to prepare the extract on a larger scale to enable further clinical testing. From this point, Connaught would play a fundamental role in the insulin story, although its historical significance has been minimized and often misunderstood, overshadowed by the weight of the primary discovery, as well as by the critical contributions of Eli Lilly and Co. This article focuses on Connaught’s intimate involvement in the history of insulin from January 1922 through the summer of 1924, tracing the challenges and innovations of developing larger-scale production methods, the establishment and expansion of Canadian insulin production capacity, and the key role the labs played in spearheading the global distribution of insulin.

Historical note: Horace Dobell's report of cystic fibrosis in 1872

In 1872, Horace Dobell, a respected Victorian physician, described the key features of cystic fibrosis, with pancreatic insufficiency, pulmonary disease, and familial recurrence, plus effective treatment of steatorrhea with a pancreatic extract, and was aware of the combination of digestive and pulmonary disease in the pediatric population! This antedated modern descriptions of the full disorder by over 60 years.

Live enteroviruses, but not other viruses, detected in human pancreas at the onset of type 1 diabetes in the DiViD study

Aims/hypothesisEnterovirus (EV) infection of pancreatic islet cells is one possible factor contributing to type 1 diabetes development. We have reported the presence of EV genome by PCR and of EV proteins by immunohistochemistry in pancreatic sections. Here we explore multiple human virus species in the Diabetes Virus Detection (DiViD) study cases using innovative methods, including virus passage in cell cultures.MethodsSix recent-onset type 1 diabetes patients (age 24–35) were included in the DiViD study. Minimal pancreatic tail resection was performed under sterile conditions. Eleven live cases (age 43–83) of pancreatic carcinoma without diabetes served as control cases. In the present study, we used EV detection methods that combine virus growth in cell culture, gene amplification and detection of virus-coded proteins by immunofluorescence. Pancreas homogenates in cell culture medium were incubated with EV-susceptible cell lines for 3 days. Two to three blind passages were performed. DNA and RNA were extracted from both pancreas tissue and cell cultures. Real-time PCR was used for detecting 20 different viral agents other than EVs (six herpesviruses, human polyomavirus [BK virus and JC virus], parvovirus B19, hepatitis B virus, hepatitis C virus, hepatitis A virus, mumps, rubella, influenza A/B, parainfluenza 1–4, respiratory syncytial virus, astrovirus, norovirus, rotavirus). EV genomes were detected by endpoint PCR using five primer pairs targeting the partially conserved 5′ untranslated region genome region of the A, B, C and D species. Amplicons were sequenced. The expression of EV capsid proteins was evaluated in cultured cells using a panel of EV antibodies.ResultsSamples from six of six individuals with type 1 diabetes (cases) and two of 11 individuals without diabetes (control cases) contained EV genomes (p<0.05). In contrast, genomes of 20 human viruses other than EVs could be detected only once in an individual with diabetes (Epstein–Barr virus) and once in an individual without diabetes (parvovirus B19). EV detection was confirmed by immunofluorescence of cultured cells incubated with pancreatic extracts: viral antigens were expressed in the cytoplasm of approximately 1% of cells. Notably, infection could be transmitted from EV-positive cell cultures to uninfected cell cultures using supernatants filtered through 100 nm membranes, indicating that infectious agents of less than 100 nm were present in pancreases. Due to the slow progression of infection in EV-carrying cell cultures, cytopathic effects were not observed by standard microscopy but were recognised by measuring cell viability. Sequences of 5′ untranslated region amplicons were compatible with EVs of the B, A and C species. Compared with control cell cultures exposed to EV-negative pancreatic extracts, EV-carrying cell cultures produced significantly higher levels of IL-6, IL-8 and monocyte chemoattractant protein-1 (MCP1).Conclusions/interpretationSensitive assays confirm that the pancreases of all DiViD cases contain EVs but no other viruses. Analogous EV strains have been found in pancreases of two of 11 individuals without diabetes. The detected EV strains can be passaged in series from one cell culture to another in the form of poorly replicating live viruses encoding antigenic proteins recognised by multiple EV-specific antibodies. Thus, the early phase of type 1 diabetes is associated with a low-grade infection by EVs, but not by other viral agents. Graphical abstract

WS04.03 Pulmonary impacts of the long-term use of proton pump inhibitors to potentialise the effect of pancreatic extracts in children with cystic fibrosis

WS04.03 Pulmonary impacts of the long-term use of proton pump inhibitors to potentialise the effect of pancreatic extracts in children with cystic fibrosis

La découverte de l’insuline

The first therapeutic benefits of insulin were recorded after the injection of pancreatic extract, given on January 23, 1922 in Toronto to a 14-year-old teenager. Until then, type I diabetes was always fatal, within weeks or months; the fatal outcome being delayed only at the cost of a drastic low-calorie diet. In previous decades, the importance of the pancreas in the development of diabetes had been pointed out, but all attempts to use a pancreatic extract had failed. It is with the objective of "neutralizing" the destructive effects of pancreatic juice (proteolytic) that the isolation of insulin was carried out by a research team which was totally improbable since it was headed by an orthopedic surgeon, Frederick Banting and a 22-year-old stagiaire, Charles Best. Their work was carried out in the university physiology laboratory of John Macleod and their outcome was made possible thanks to the skills of James Collip who purified the insulin preparation. Scientific reality invites us to emphasize that, Banting works, based on a wrong hypothesis, drew towards an historical discovery. Very quickly recognized as of major importance for medicine, the discovery was greeted by the attribution of the Nobel Prize in 1923. For a hundred years, insulin has not ceased to be an essential drug for tens of millions of patients in the world, but it has been a motor for scientific research: innovation in galenic pharmacy and biopharmacy, in fundamental chemistry as a subject for the study of the structure, analysis and synthesis of proteins, and finally, as a motor for the development of biotechnologies, since insulin was the first drug prepared by DNA-recombinant technology, and marketed in 1982.