{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002237","ANALYSIS_ID":"AN003650","VERSION":"1","CREATED_ON":"August 2, 2022, 8:19 am"},

"PROJECT":{"PROJECT_TITLE":"Metabolomic analysis of brain cortex from neuronal specific Depdc5 knockout in fed and fasting states","PROJECT_TYPE":"LCMS based comprehensive hydrophilic metabolites profiling","PROJECT_SUMMARY":"We performed a metabolomic analysis of the brain cortex from neuronal-specific Depdc5 knockout in fed and fasting states.","INSTITUTE":"Northwestern University Feinberg School of Medicine","DEPARTMENT":"Medicine","LABORATORY":"Chandel Lab","LAST_NAME":"Chandel","FIRST_NAME":"Navdeep","ADDRESS":"303 E Superior St, Chicago, Illinois, 60611, USA","EMAIL":"nav@northwestern.edu","PHONE":"3125032549","FUNDING_SOURCE":"NIA","PUBLICATIONS":"DEPDC5-dependent mTORC1 signaling mechanisms are critical for the anti-seizure effects of acute fasting","CONTRIBUTORS":"Christopher J. Yuskaitis, Jinita B. Modasia, Sandra Schroetter, Leigh-Ana Rossitto, Karenna Groff, Christopher Morici, Divakar S. Mithal, Ram P. Chakrabarty, Navdeep S. Chandel, Brendan D. Manning, Mustafa Sahin"},

"STUDY":{"STUDY_TITLE":"Metabolomic analysis of brain cortex from neuronal specific Depdc5 knockout in fed and fasting state","STUDY_TYPE":"Depdc5 KO fed vs. fasted comparison","STUDY_SUMMARY":"Caloric restriction and acute fasting are known to reduce seizures but through unclear mechanisms. In this study, we demonstrate that mTORC1 signaling is reduced after acute fasting of mice. In neurons, mTORC1 is most sensitive to withdrawal of leucine, arginine, and glutamine, which is dependent on DEPDC5. We performed metabolomic analysis of brain cortex from neuronal specific Depdc5 knockout in fed and fasting state. The Depdc5 neuronal specific knockout mice are resistant to sensing significant fluctuations in brain amino acid levels after fasting. These results establish that acute fasting reduces seizure susceptibility in a DEPDC5-dependent manner.","INSTITUTE":"Northwestern University Feinberg School of Medicine","DEPARTMENT":"Medicine","LABORATORY":"Chandel Lab","LAST_NAME":"Chandel","FIRST_NAME":"Navdeep","ADDRESS":"303 E Superior St, Chicago, Illinois, 60611, USA","EMAIL":"nav@northwestern.edu","PHONE":"3125032549","NUM_GROUPS":"4","TOTAL_SUBJECTS":"40","NUM_MALES":"20","NUM_FEMALES":"20","PUBLICATIONS":"DEPDC5-dependent mTORC1 signaling mechanisms are critical for the anti-seizure effects of acute fasting"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"166-3",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Control"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-10.raw"}
},
{
"Subject ID":"-",
"Sample ID":"166-4",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Control"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-11.raw"}
},
{
"Subject ID":"-",
"Sample ID":"166-5",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Fasted"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-12.raw"}
},
{
"Subject ID":"-",
"Sample ID":"166-6",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Fasted"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-13.raw"}
},
{
"Subject ID":"-",
"Sample ID":"167-7",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Control"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-14.raw"}
},
{
"Subject ID":"-",
"Sample ID":"169-2",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Control"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-15.raw"}
},
{
"Subject ID":"-",
"Sample ID":"169-3",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Fasted"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-16.raw"}
},
{
"Subject ID":"-",
"Sample ID":"169-5",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Control"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-17.raw"}
},
{
"Subject ID":"-",
"Sample ID":"189-2",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Control"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-18.raw"}
},
{
"Subject ID":"-",
"Sample ID":"189-3",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Control"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-19.raw"}
},
{
"Subject ID":"-",
"Sample ID":"190-4",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Fasted"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-20.raw"}
},
{
"Subject ID":"-",
"Sample ID":"190-5",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Fasted"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-21.raw"}
},
{
"Subject ID":"-",
"Sample ID":"191-1",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Control"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-22.raw"}
},
{
"Subject ID":"-",
"Sample ID":"191-3",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Control"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-23.raw"}
},
{
"Subject ID":"-",
"Sample ID":"192-1",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Fasted"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-24.raw"}
},
{
"Subject ID":"-",
"Sample ID":"192-2",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Control"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-25.raw"}
},
{
"Subject ID":"-",
"Sample ID":"192-4",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Control"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-26.raw"}
},
{
"Subject ID":"-",
"Sample ID":"192-5",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Control"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-27.raw"}
},
{
"Subject ID":"-",
"Sample ID":"192-6",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Fasted"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-28.raw"}
},
{
"Subject ID":"-",
"Sample ID":"248-1",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Fasted"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-29.raw"}
},
{
"Subject ID":"-",
"Sample ID":"248-6",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Fasted"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-30.raw"}
},
{
"Subject ID":"-",
"Sample ID":"248-7",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Control"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-31.raw"}
},
{
"Subject ID":"-",
"Sample ID":"600-1",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Control"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-32.raw"}
},
{
"Subject ID":"-",
"Sample ID":"600-7",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Fasted"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-33.raw"}
},
{
"Subject ID":"-",
"Sample ID":"600-8",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Fasted"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-34.raw"}
},
{
"Subject ID":"-",
"Sample ID":"601-1",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Control"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-35.raw"}
},
{
"Subject ID":"-",
"Sample ID":"601-2",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Fasted"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-36.raw"}
},
{
"Subject ID":"-",
"Sample ID":"601-4",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Fasted"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-37.raw"}
},
{
"Subject ID":"-",
"Sample ID":"601-5",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Fasted"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-38.raw"}
},
{
"Subject ID":"-",
"Sample ID":"609-6",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Control"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-39.raw"}
},
{
"Subject ID":"-",
"Sample ID":"611-5",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Fasted"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-40.raw"}
},
{
"Subject ID":"-",
"Sample ID":"625-1",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Control"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-44.raw"}
},
{
"Subject ID":"-",
"Sample ID":"627-6",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Fasted"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-45.raw"}
},
{
"Subject ID":"-",
"Sample ID":"627-7",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Fasted"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-46.raw"}
},
{
"Subject ID":"-",
"Sample ID":"986-3",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Fasted"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-65.raw"}
},
{
"Subject ID":"-",
"Sample ID":"986-5",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Fasted"},
"Additional sample data":{"Sex":"F","RAW_FILE_NAME":"Nav-Ram-20200224-66.raw"}
},
{
"Subject ID":"-",
"Sample ID":"993-1",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Control"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-77.raw"}
},
{
"Subject ID":"-",
"Sample ID":"993-10",
"Factors":{"Genotype":"Depdc5 KO","Treatment":"Control"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-78.raw"}
},
{
"Subject ID":"-",
"Sample ID":"993-4",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Control"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-79.raw"}
},
{
"Subject ID":"-",
"Sample ID":"993-5",
"Factors":{"Genotype":"Depdc5 WT","Treatment":"Fasted"},
"Additional sample data":{"Sex":"M","RAW_FILE_NAME":"Nav-Ram-20200224-80.raw"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Mouse cerebral cortex was rapidly dissected and flash frozen.","SAMPLE_TYPE":"Brain","STORAGE_CONDITIONS":"-80℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"Group housed mice, 6-9 weeks old Depdc5cc+ mice and their littermate controls, were randomized to fed or fasted conditions. Mice were weighed, followed by food removal at 1 pm for 24hr with ad lib water access. Twenty-four hours post-fasting, mice were weighed, followed by experimentation."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Mouse cortical lysate samples were rapidly isolated, and flash frozen in liquid nitrogen. Soluble metabolites were extracted directly from tissue using cold methanol/water (80/20, v/v) at approximately 1μL per 50μg of tissue, followed by ultrasonication (Branson Sonifier 250) for 15 s. Homogenized samples were incubated at −80 °C to precipitate proteins and subsequently centrifuged at 18,000xg for 20 min at 4 °C to pellet the debris. The supernatants containing soluble metabolites were collected in new tubes and evaporated to dryness using a SpeedVac concentrator (Thermo Savant). Next, the metabolites were reconstituted in acetonitrile/water (60/40, v/v), vortex-mixed, and centrifuged at 18,000xg for 30 min at 4°C to remove debris."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Samples were analyzed by High-Performance Liquid Chromatography and High-Resolution Mass Spectrometry and Tandem Mass Spectrometry (HPLC-MS/MS). Specifically, system consisted of a Thermo Q-Exactive in line with an electrospray source and an Ultimate3000 (Thermo) series HPLC consisting of a binary pump, degasser, and auto-sampler outfitted with a Xbridge Amide column (Waters; dimensions of 3.0 mm × 100 mm and a 3.5 µm particle size). The mobile phase A contained 95% (vol/vol) water, 5% (vol/vol) acetonitrile, 10 mM ammonium hydroxide, 10 mM ammonium acetate, pH = 9.0; B was 100% Acetonitrile. The gradient was as following: 0 min, 15% A; 2.5 min, 30% A; 7 min, 43% A; 16 min, 62% A; 16.1-18 min, 75% A; 18-25 min, 15% A with a flow rate of 150 μL/min. The capillary of the ESI source was set to 275 °C, with sheath gas at 35 arbitrary units, auxiliary gas at 5 arbitrary units and the spray voltage at 4.0 kV. In positive/negative polarity switching mode, an m/z scan range from 60 to 900 was chosen and MS1 data was collected at a resolution of 70,000. The automatic gain control (AGC) target was set at 1 × 106 and the maximum injection time was 200 ms. The top 5 precursor ions were subsequently fragmented, in a data-dependent manner, using the higher energy collisional dissociation (HCD) cell set to 30% normalized collision energy in MS2 at a resolution power of 17,500. Besides matching m/z, metabolites are identified by matching either retention time with analytical standards and/or MS2 fragmentation pattern. Data acquisition and analysis were carried out by Xcalibur 4.1 software and Tracefinder 4.1 software, respectively (both from Thermo Fisher Scientific).","CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Thermo Dionex Ultimate 3000","COLUMN_NAME":"Waters Xbridge Amide (100 x 3mm, 3.5um)"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"UNSPECIFIED","MS_COMMENTS":"The capillary of the ESI source was set to 275 °C, with sheath gas at 35 arbitrary units, auxiliary gas at 5 arbitrary units and the spray voltage at 4.0 kV. In positive/negative polarity switching mode, an m/z scan range from 60 to 900 was chosen and MS1 data was collected at a resolution of 70,000. The automatic gain control (AGC) target was set at 1 × 106 and the maximum injection time was 200 ms. The top 5 precursor ions were subsequently fragmented, in a data-dependent manner, using the higher energy collisional dissociation (HCD) cell set to 30% normalized collision energy in MS2 at a resolution power of 17,500. Besides matching m/z, metabolites are identified by matching either retention time with analytical standards and/or MS2 fragmentation pattern. Data acquisition and analysis were carried out by Xcalibur 4.1 software and Tracefinder 4.1 software, respectively (both from Thermo Fisher Scientific)."},

"MS_METABOLITE_DATA":{
"Units":"Peak area",

"Data":[{"Metabolite":"(S)-4-hydroxymandelonitrile +","166-3":"0","166-4":"0","166-5":"0","166-6":"0","167-7":"0","169-2":"0","169-3":"0","169-5":"0","189-2":"0","189-3":"0","190-4":"0","190-5":"0","191-1":"0","191-3":"0","192-1":"0","192-2":"0","192-4":"0","192-5":"0","192-6":"0","248-1":"0","248-6":"319156","248-7":"0","600-1":"0","600-7":"0","600-8":"0","601-1":"0","601-2":"0","601-4":"0","601-5":"0","609-6":"0","611-5":"0","625-1":"0","627-6":"849101","627-7":"0","986-3":"0","986-5":"0","993-1":"0","993-10":"0","993-4":"724268","993-5":"0"},{"Metabolite":"1,3-bisphospho-D-glycerate","166-3":"0","166-4":"0","166-5":"0","166-6":"0","167-7":"0","169-2":"0","169-3":"0","169-5":"0","189-2":"0","189-3":"0","190-4":"0","190-5":"0","191-1":"0","191-3":"0","192-1":"0","192-2":"0","192-4":"0","192-5":"0","192-6":"0","248-1":"0","248-6":"0","248-7":"0","600-1":"0","600-7":"0","600-8":"0","601-1":"0","601-2":"0","601-4":"0","601-5":"0","609-6":"0","611-5":"0","625-1":"0","627-6":"0","627-7":"0","986-3":"0","986-5":"0","993-1":"0","993-10":"0","993-4":"0","993-5":"0"},{"Metabolite":"1-methylnicotinamide+","166-3":"8908557","166-4":"4946816","166-5":"11303747","166-6":"6749548","167-7":"12762234","169-2":"6324035","169-3":"29390747","169-5":"7732310","189-2":"13675285","189-3":"11281729","190-4":"14718309","190-5":"14191565","191-1":"16007905","191-3":"10151729","192-1":"19741024","192-2":"11582490","192-4":"23608923","192-5":"17962193","192-6":"17761206","248-1":"10561553","248-6":"24494697","248-7":"19465787","600-1":"12636866","600-7":"11653651","600-8":"16402958","601-1":"12693018","601-2":"12473923","601-4":"10704353","601-5":"19416339","609-6":"9419760","611-5":"19375886","625-1":"13756395","627-6":"19624527","627-7":"14022054","986-3":"17698375","986-5":"15685109","993-1":"7305138","993-10":"12341320","993-4":"4968770","993-5":"9247407"},{"Metabolite":"2-(Formamido)-N1-(5-phospho-D-ribosyl)acetamidine","166-3":"230731","166-4":"193467","166-5":"452407","166-6":"118743","167-7":"36335","169-2":"165583","169-3":"127815","169-5":"394462","189-2":"198922","189-3":"346323","190-4":"361968","190-5":"333167","191-1":"404106","191-3":"339013","192-1":"167291","192-2":"259599","192-4":"199981","192-5":"159437","192-6":"145418","248-1":"168021","248-6":"60158","248-7":"139077","600-1":"142953","600-7":"116089","600-8":"284446","601-1":"230180","601-2":"421825","601-4":"137850","601-5":"96152","609-6":"0","611-5":"119777","625-1":"0","627-6":"84352","627-7":"148395","986-3":"178498","986-5":"147671","993-1":"175996","993-10":"293734","993-4":"564315","993-5":"159046"},{"Metabolite":"2,3-bisphospho-D-glycerate","166-3":"95111","166-4":"0","166-5":"334521","166-6":"0","167-7":"0","169-2":"109030","169-3":"0","169-5":"135494","189-2":"72825","189-3":"309281","190-4":"448049","190-5":"281233","191-1":"165818","191-3":"303337","192-1":"42795","192-2":"63778","192-4":"0","192-5":"338371","192-6":"0","248-1":"24761","248-6":"173611","248-7":"214699","600-1":"38219","600-7":"0","600-8":"231240","601-1":"397041","601-2":"283479","601-4":"236899","601-5":"295129","609-6":"261276","611-5":"37728","625-1":"98247","627-6":"376774","627-7":"106453","986-3":"29039","986-5":"134427","993-1":"525874","993-10":"1391295","993-4":"1733466","993-5":"650639"},{"Metabolite":"2/3-phosphoglycerate","166-3":"13769053","166-4":"5583247","166-5":"17003653","166-6":"4308986","167-7":"9277423","169-2":"14197341","169-3":"7937463","169-5":"11148967","189-2":"18746845","189-3":"16819260","190-4":"21504800","190-5":"13023760","191-1":"14781684","191-3":"16200171","192-1":"10272003","192-2":"15677418","192-4":"14509478","192-5":"20665162","192-6":"10554203","248-1":"9856833","248-6":"13045492","248-7":"18007156","600-1":"15355324","600-7":"11782308","600-8":"18660547","601-1":"20340262","601-2":"18438833","601-4":"22826163","601-5":"14022526","609-6":"15785189","611-5":"12752357","625-1":"13573398","627-6":"15379028","627-7":"10986853","986-3":"13007319","986-5":"16830380","993-1":"19558855","993-10":"44266252","993-4":"14237148","993-5":"30762599"},{"Metabolite":"2-aminobutanoate","166-3":"1602570","166-4":"762441","166-5":"9412331","166-6":"11304617","167-7":"637062","169-2":"3498970","169-3":"17444152","169-5":"1477135","189-2":"4868683","189-3":"1470102","190-4":"7573258","190-5":"5773124","191-1":"1931467","191-3":"3138867","192-1":"6040834","192-2":"1254327","192-4":"2267403","192-5":"2139812","192-6":"6541257","248-1":"7758883","248-6":"14474926","248-7":"1403497","600-1":"2030213","600-7":"7258168","600-8":"14799553","601-1":"2029577","601-2":"6369479","601-4":"7111040","601-5":"18002458","609-6":"963782","611-5":"12351253","625-1":"4339770","627-6":"7938268","627-7":"12932597","986-3":"13512899","986-5":"7822871","993-1":"6665165","993-10":"2356955","993-4":"4449515","993-5":"8479675"},{"Metabolite":"2-aminoisobutanoate","166-3":"94346","166-4":"521272","166-5":"424157","166-6":"341919","167-7":"1017567","169-2":"574231","169-3":"526325","169-5":"376630","189-2":"105787","189-3":"221926","190-4"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}

}