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Gangadhara"],"software":["Not available"],"tissue":["Pancreatic Islet","Islet Of Langerhans"],"sample_protocol":["Induction of MetS Healthy male C57BL/6J mice, were maintained under controlled conditions (25 ± 2°C, 12-h light/dark cycle), as per the guidelines of Committee for Control and Supervision of Experiments on Animals India. The experiments were approved by the Institutional Animal Ethics Committee of St. Aloysius College (Autonomous) Mangalore, India (Sanction No: SAC/IAEC/05/2019, 28 January 2019). One-month-old healthy male C57BL/6J mice weighing 18–20 g was randomly divided into control (n=10) and HFSC (n=10) groups. The control group was given a regular diet, whereas the HFSC group was fed a specially formulated HFSC diet. Both groups were given 5 g/d of their respective diets, per Indian National Science Academy regulations (2000), with ad libitum access to water. These conditions were maintained for 150 d. The control and HFSC diet were formulated as per D Souza et al., 2016 . Preparation of pancreatic islets At the end of 150 d, both HFSC and control diet-fed mice were euthanized by cervical dislocation . The pancreas was dissected, and the splenic region was separated and washed with ice-cold Hank's balanced salt solution (HBSS) under aseptic conditions. The pancreas was maintained in a bicarbonate-buffered medium at physiological pH , with excess fat tissues manually removed. Collagenase type IV (Sigma-Aldrich, St. Louis, MO, USA) was injected at a concentration of 1 mg/mL in HBSS to the dissected pancreas repeatedly to disrupt the connective tissue. The finely chopped pancreatic tissues were further incubated with collagenase in a sterile glass tube at 37 ± 2℃ with vigorous shaking for 15 min. After digestion, the suspension was filtered to separate cell debris and fat, followed by washing with buffer and centrifugation at 500 × g for 60 s at room temperature. The islet suspension was then transferred to a sterile Petri dish. Pancreatic islets appearing as yellow-sphere-like structures, were collected using sterile Pasteur pipettes under aseptic conditions while being observed through a dissection microscope. .  Isolation of proteins from pancreatic islets Pancreatic islets were subjected to two rounds of washing with ice-cold PBS, followed by centrifugation at 1200 rpm for 3 min. The resulting pellet was carefully transferred to a 1.5 mL safe-lock tube. After a final wash, the pellet was vigorously spun down in a microcentrifuge and aspirated to remove the remaining liquid. Subsequently, 50 L of Rapigest extraction buffer (Waters, Manchester, UK)  supplemented with protease inhibitors was added to the pellet, and the mixture was vigorously shaken. The reaction proceeded at room temperature for 10 min. Freeze-thaw cycles were then performed in liquid nitrogen, repeated twice. The resulting cell suspension was subjected to centrifugation at 15,000 rpm for 30 min at 4°C to obtain the supernatant containing protein, which was collected in a fresh tube. The isolated proteins underwent desalting through buffer exchange using Amicon Ultra 0.5 mL centrifugal filters  (Merck Millipore, Billerica, MA, USA)with a 3 kDa molecular weight cutoff (MWCO), employing a 50 mM ammonium bicarbonate buffer. Following buffer exchange, the protein samples were quantified using the Pierce bicinchoninic acid kit (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s protocol and subsequently diluted to a uniform concentration of 1 mg/mL. The protein samples from both HFSC and control groups were reduced using 100 mM 1,4-Dithiothreitol (Sigma-Aldrich), followed by alkylation with 200 mM iodoacetamide (Sigma-Aldrich). Subsequently, overnight digestion was carried out using MS-grade trypsin (Sigma-Aldrich) at a ratio of 1:25."],"repository":["Pride"],"labhead":["Asha Abraham"],"instrument_platform":[""],"labhead_affiliation":["Department of Biotechnology  St Aloysius (Deemed to be University)  Mangalore – 575 003, Karnataka, India."],"submission_type":["COMPLETE"],"publication":["10.6019/PXD057058"],"submitter_affiliation":["St Aloysius (Deemed to be University), Mangaluru"],"labhead_mail":["drashaabraham@staloysius.edu.in"],"technology_type":["Mass Spectrometry","Data-independent acquisition"],"disease":["Type 2 Diabetes Mellitus"],"submitter_keywords":["Lc-msms","Mouse","Pancreatic islet"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD057058"],"modification":[""],"quantification_method":[""],"data_protocol":["Protein Profiling using ESI-nanoLC-MS/MS Liquid chromatography analyses were carried out using a nanoACQUITY UPLC® system (Waters) coupled with MassLynx4.1 SCN781 software for data acquisition. A two-solvent system consisting of solvent A (0.1% formic acid in water) and solvent B (0.1% formic acid in acetonitrile) was applied. For trapping purposes, a Symmetry® C18 column (180 μm × 20 mm, 5 μm, Waters) was employed, followed by a 75 μm × 200 mm HSS T3 C18 column (1.8 μm, Waters) for separation. Each sample was injected in duplicate, with an injection volume of 0.5 µl. The flow rate was maintained at 300 nL/min, with a column temperature of 35°C. Analyses were conducted in reverse phase mode, with an autosampler temperature of 4°C. The overall liquid chromatography gradient run time was 90 minutes. Mass spectrometry analysis was conducted using ion mobility enabled separation on a Synapt G2 High Definition MS™ System (HDMSE System) by Waters. Calibration was performed using sodium iodide, and the acquisition mode was set to electrospray ionization (ESI) in positive polarity. Online mass correction utilized Leucine encephalin in positive ion mode with an m/z of 556.2766. Nano electrospray ionization (ESI) capillary voltage was set at 3.4 KV, with sample cone and extraction cone voltages set to 40 V and 4 V, respectively. Nitrogen was used as the IMS gas with a flow rate of 90 mL/min. IMS T-Wave™ pulse height and velocity were set to 40 V and 800 m/s, respectively, with IMS voltage operated at 8 V and 20 V. The mode of operation was resolution mode, and data were acquired in continuum format. Collision energy ranged from 20 eV to 45 eV.  Analysis of Proteomics Data Progenesis QI for Proteomics software (Non-linear Dynamics, Newcastle upon Tyne, UK) was utilized for the analysis of proteomics data. The Mus musculus protein database UniProt entries were used for peptide annotation. The default protocol provided on the Progenesis website was followed for data analysis. The peak picking of data was performed using an automatic sensitivity method based on a noise estimation algorithm, with a maximum ion charge of 20. A false discovery rate of 4% was used for the search, and the identification of at least one unique peptide was required for protein identification. Fixed and variable modifications were set for cysteine carbamidomethylation and methionine oxidation, respectively. Trypsin digestion parameter was set to one missed cleavage for specificity during database search. Normalized intensity values obtained from three biological replicates were used for label-free quantification analysis."],"omics_type":["Proteomics"],"species":["Mus Musculus (mouse)"],"submitter_mail":["vijayalakshmi@staloysius.edu.in"],"submitter_country":["India"],"doi":["10.6019/PXD057058"],"additional_accession":[]},"is_claimable":false,"name":"Proteomic analysis of mice pancreatic islet fed a high fat simple carbohydrate diet","description":"This study aims to elucidate the molecular mechanisms linking dietary stress from a high-fat simple carbohydrate (HFSC) diet to an elevated risk of pancreatic cancer. Using label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS), we performed a comprehensive proteomic analysis of collagenase-digested pancreatic islets from HFSC-fed mice.  The study identified a significant upregulation and downregulation of 33 and 34 proteins respectively in HFSC group compared to the control group. The results revealed substantial alterations in the expression of proteins involved in key biological processes such as metabolic stress, protein synthesis, and endoplasmic reticulum (ER) stress, all of which are crucial for maintaining cellular homeostasis under dietary stress. Notably, the study identified several potential biomarkers, including MANF, VCP, HYOU1, and PRDX1, which are intricately linked to pancreatic function and the progression of cancer. These biomarkers play pivotal roles in managing cellular stress responses, protein folding, and metabolic regulation, offering insights into how a HFSC diet contributes to pancreatic carcinogenesis. This research highlights the significant impact of dietary factors on pancreatic cancer development and underscores the potential of these biomarkers as therapeutic targets for early detection and treatment, ultimately aiming to improve patient outcomes in PDAC.","dates":{"publication":"2026-04-22","submission":"2024-10-22"},"accession":"PXD057058","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:1131","NEWT:35795","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NEWT:1334565","NEWT:625351","NEWT:258594","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:247480","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:475174","NEWT:1599","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:12215","NEWT:1590","NEWT:10034","NEWT:33114","NEWT:1596","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:294671","NEWT:376619","NEWT:1748418","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:3767","NEWT:1589","NEWT:135622","NEWT:4615","NEWT:672127","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656064","NEWT:1582","NEWT:3760","NEWT:1584","NEWT:2086595","NEWT:36630","NEWT:1585","NEWT:383379","NEWT:381198","NEWT:418106","NCBITaxon:12721","NEWT:166340","NEWT:10029","NEWT:913645","NEWT:641809","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:3357","NEWT:7719","NEWT:29073","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:1562698","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:108061","NEWT:60711","NEWT:2020713","NEWT:224308","NEWT:3347","NEWT:49928","NEWT:3349","NEWT:160621","NEWT:2502409","NEWT:212790","NEWT:110368","NEWT:1916073","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:410658","NEWT:5516","NEWT:44688","NEWT:44689","NEWT:370038","NEWT:498211","NEWT:345076","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:1310173","NEWT:200795","NEWT:2579791","NEWT:44685","NEWT:161934","NEWT:156587","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:260499","NEWT:410661","NEWT:329854","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:2649329","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:150808","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:3714","NCBITaxon:37162","NEWT:270643","NEWT:3715","NEWT:2065263","NEWT:199225","NEWT:177437","NEWT:717966","NEWT:10891","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:28174","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:244366","NEWT:1246791","NEWT:176597","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:52299","NEWT:8165","NEWT:96794","NEWT:10886","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:926571","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:1955249","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:2714896","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:1608994","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:56636","NEWT:2499525","NEWT:1122981","NEWT:2853422","NEWT:1122984","NEWT:487","NEWT:1538553","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:45972","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:1502","NEWT:128017","NEWT:2836","NEWT:376686","NEWT:237610","NEWT:95486","EFO:0001352","NEWT:119562","NEWT:1883446","NEWT:31345","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:1509","NEWT:491","NEWT:460523","NEWT:45954","NEWT:235443","NEWT:1579","NEWT:1757312","NEWT:85694","NEWT:109786","NEWT:1922344","NEWT:108458","NEWT:5936","NEWT:85692","NEWT:321967","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:279010","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:1116234","NEWT:415978","NEWT:235455","NEWT:1255228","NEWT:410289","NEWT:191701","NEWT:373153","NEWT:1640278","NEWT:9054","NEWT:472","NEWT:940825","NEWT:29459","NEWT:529507","NEWT:1071661","NEWT:28128","NEWT:470","NEWT:41122","NEWT:5911","NEWT:1328300","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:11723","NEWT:445","NEWT:153009","NEWT:261756","NEWT:166314","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:1871049","NEWT:215402","NEWT:1547","NEWT:1290454","NEWT:3726","NEWT:906914","NEWT:81231006","NEWT:30455","NEWT:28141","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:3728","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:548","NEWT:150475","NEWT:267872","NEWT:381666","NEWT:172269","NEWT:9534","NEWT:2126348","NEWT:5180","NEWT:256737","NEWT:299467","NEWT:9541","NEWT:51350","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:542","NEWT:2903","NEWT:185579","NEWT:872227","NEWT:13076","NEWT:2587410","NEWT:207340","NEWT:1006581","NEWT:8673","NEWT:33940","NEWT:2126337","NEWT:550","NEWT:253237","NEWT:10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