{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001407","ANALYSIS_ID":"AN002351","VERSION":"1","CREATED_ON":"June 22, 2020, 12:08 pm"},

"PROJECT":{"PROJECT_TITLE":"Environmental chemical burden in metabolic tissues and systemic biological pathways in adolescent bariatric surgery patients: A pilot untargeted metabolomic approach","PROJECT_TYPE":"Pilot Study","PROJECT_SUMMARY":"Background: Advances in untargeted metabolomic technologies have great potential for insight into adverse metabolic effects underlying exposure to environmental chemicals. However, important challenges need to be addressed, including how biological response corresponds to the environmental chemical burden in different target tissues. Aim: We performed a pilot study using state-of-the-art ultra-high-resolution mass spectrometry (UHRMS) to characterize the burden of lipophilic persistent organic pollutants (POPs) in metabolic tissues and associated alterations in the plasma metabolome. Methods: We studied 11 adolescents with severe obesity at the time of bariatric surgery. We measured 18 POPs that can act as endocrine and metabolic disruptors (i.e. 2 dioxins, 11 organochlorine compounds [OCs] and 5 polybrominated diphenyl ethers [PBDEs]) in visceral and subcutaneous abdominal adipose tissue (vAT and sAT), and liver samples using gas chromatography with UHRMS. Biological pathways were evaluated by measuring the plasma metabolome using high-resolution metabolomics. Network and pathway enrichment analysis assessed correlations between the tissue-specific burden of three frequently detected POPs (i.e. p,p’-dichlorodiphenyldichloroethene [DDE], hexachlorobenzene [HCB] and PBDE-47) and plasma metabolic pathways. Results: Concentrations of 4 OCs and 3 PBDEs were quantifiable in at least one metabolic tissue for >80% of participants. All POPs had the highest median concentrations in adipose tissue, especially sAT, except for PBDE-154, which had comparable average concentrations across all tissues. Pathway analysis showed high correlations between tissue-specific POPs and metabolic alterations in pathways of amino acid metabolism, lipid and fatty acid metabolism, and carbohydrate metabolism. Conclusions: Most of the measured POPs appear to accumulate preferentially in adipose tissue compared to liver. Findings of plasma metabolic pathways potentially associated with tissue-specific POPs concentrations merit further investigation in larger populations. Keywords: persistent organic pollutants, adipose tissue, liver, bariatric surgery, exposome, high-resolution metabolomics","INSTITUTE":"Icahn School of Medicine at Mount Sinai","DEPARTMENT":"Environmental Medicine and Public Health","LABORATORY":"High Resolution Exposomics Research Group","LAST_NAME":"Walker","FIRST_NAME":"Douglas","ADDRESS":"One Gustave L. Levy Place, Box 1057, New York, NY 10029","EMAIL":"douglas.walker@mssm.edu","PHONE":"212-241-9891","FUNDING_SOURCE":"NIEHS: R21ES028903, R21ES029328, R21ES029681, R01ES029944, R01ES030364, U2CES026561, U2CES030163, P30ES023515, P30 ES019776, P30ES007048, P01ES022845, R01ES024946; EPA: RD-83544101","PROJECT_COMMENTS":"This upload is 2 of 2 and includes untargeted plasma metabolomics results. Part 1 included targeted levels of organic pollutants","PUBLICATIONS":"Valvi D, Walker DI, Inge T, Bartell SM, Jenkins T, Helmrath M, Ziegler TR, La Merrill MA, Eckel SP, Conti D, Liang Y, Jones DP, McConnell R, Chatzi L. (2020). Environmental chemical burden in metabolic tissues and systemic biological pathways in adolescent bariatric surgery patients: A pilot untargeted metabolomic approach. Environment International. In Press.","CONTRIBUTORS":"Valvi D, Walker DI, Inge T, Bartell SM, Jenkins T, Helmrath M, Ziegler TR, La Merrill MA, Eckel SP, Conti D, Liang Y, Jones DP, McConnell R, Chatzi L"},

"STUDY":{"STUDY_TITLE":"Environmental chemical burden in metabolic tissues and systemic biological pathways in adolescent bariatric surgery patients: A pilot untargeted metabolomic approach","STUDY_TYPE":"Subcutaneous adipose tissue (AT); Visceral AT; Liver Tissue; Plasma","STUDY_SUMMARY":"Background: Advances in untargeted metabolomic technologies have great potential for insight into adverse metabolic effects underlying exposure to environmental chemicals. However, important challenges need to be addressed, including how biological response corresponds to the environmental chemical burden in different target tissues. Aim: We performed a pilot study using state-of-the-art ultra-high-resolution mass spectrometry (UHRMS) to characterize the burden of lipophilic persistent organic pollutants (POPs) in metabolic tissues and associated alterations in the plasma metabolome. Methods: We studied 11 adolescents with severe obesity at the time of bariatric surgery. We measured 18 POPs that can act as endocrine and metabolic disruptors (i.e. 2 dioxins, 11 organochlorine compounds [OCs] and 5 polybrominated diphenyl ethers [PBDEs]) in visceral and subcutaneous abdominal adipose tissue (vAT and sAT), and liver samples using gas chromatography with UHRMS. Biological pathways were evaluated by measuring the plasma metabolome using high-resolution metabolomics. Network and pathway enrichment analysis assessed correlations between the tissue-specific burden of three frequently detected POPs (i.e. p,p’-dichlorodiphenyldichloroethene [DDE], hexachlorobenzene [HCB] and PBDE-47) and plasma metabolic pathways. Results: Concentrations of 4 OCs and 3 PBDEs were quantifiable in at least one metabolic tissue for >80% of participants. All POPs had the highest median concentrations in adipose tissue, especially sAT, except for PBDE-154, which had comparable average concentrations across all tissues. Pathway analysis showed high correlations between tissue-specific POPs and metabolic alterations in pathways of amino acid metabolism, lipid and fatty acid metabolism, and carbohydrate metabolism. Conclusions: Most of the measured POPs appear to accumulate preferentially in adipose tissue compared to liver. Findings of plasma metabolic pathways potentially associated with tissue-specific POPs concentrations merit further investigation in larger populations.","INSTITUTE":"Icahn School of Medicine at Mount Sinai","DEPARTMENT":"Environmental Medicine and Public Health","LABORATORY":"High Resolution Exposomics Research Group","LAST_NAME":"Walker","FIRST_NAME":"Doug","ADDRESS":"One Gustave L. Levy Place, Box 1057, New York, NY 10029","EMAIL":"douglas.walker@mssm.edu","PHONE":"212-241-9891","NUM_GROUPS":"1","TOTAL_SUBJECTS":"11","NUM_MALES":"1","NUM_FEMALES":"10","STUDY_COMMENTS":"Upload #1: Visceral and subcutaneous abdominal adipose tissue, liver tissue. Plasma metabolomics are in upload #2","PUBLICATIONS":"Valvi D, Walker DI, Inge T, Bartell SM, Jenkins T, Helmrath M, Ziegler TR, La Merrill MA, Eckel SP, Conti D, Liang Y, Jones DP, McConnell R, Chatzi L. (2020). Environmental chemical burden in metabolic tissues and systemic biological pathways in adolescent bariatric surgery patients: A pilot untargeted metabolomic approach. Environment International. In Press."},

"SUBJECT":{"SUBJECT_TYPE":"Human","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606","AGE_OR_AGE_RANGE":"11-20 years","GENDER":"Male and female"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"QC",
"Sample ID":"NIST_1958_1",
"Factors":{"Description":"NIST 1958"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_001","RAW_FILE_NAME":"VT_170706_003","RAW_FILE_NAME":"VT_170706_005","RAW_FILE_NAME":"VT_170706_002","RAW_FILE_NAME":"VT_170706_004","RAW_FILE_NAME":"VT_170706_006"}
},
{
"Subject ID":"QC",
"Sample ID":"q3June2014_1a",
"Factors":{"Description":"Q-Standard plasma pool"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_007","RAW_FILE_NAME":"VT_170706_009","RAW_FILE_NAME":"VT_170706_011","RAW_FILE_NAME":"VT_170706_008","RAW_FILE_NAME":"VT_170706_010","RAW_FILE_NAME":"VT_170706_012"}
},
{
"Subject ID":"QC",
"Sample ID":"q3June2014_1b",
"Factors":{"Description":"Q-Standard plasma pool"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_013","RAW_FILE_NAME":"VT_170706_015","RAW_FILE_NAME":"VT_170706_017","RAW_FILE_NAME":"VT_170706_014","RAW_FILE_NAME":"VT_170706_016","RAW_FILE_NAME":"VT_170706_013"}
},
{
"Subject ID":"QC",
"Sample ID":"chearplasma_1a",
"Factors":{"Description":"CHEAR plasma pool"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_019","RAW_FILE_NAME":"VT_170706_021","RAW_FILE_NAME":"VT_170706_023","RAW_FILE_NAME":"VT_170706_020","RAW_FILE_NAME":"VT_170706_022","RAW_FILE_NAME":"VT_170706_014"}
},
{
"Subject ID":"QC",
"Sample ID":"chearplasma_1b",
"Factors":{"Description":"CHEAR plasma pool"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_025","RAW_FILE_NAME":"VT_170706_027","RAW_FILE_NAME":"VT_170706_029","RAW_FILE_NAME":"VT_170706_026","RAW_FILE_NAME":"VT_170706_028","RAW_FILE_NAME":"VT_170706_015"}
},
{
"Subject ID":"Study_Sample",
"Sample ID":"POTR_02_Plasma",
"Factors":{"Description":"POTR_02"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_037","RAW_FILE_NAME":"VT_170706_039","RAW_FILE_NAME":"VT_170706_041","RAW_FILE_NAME":"VT_170706_038","RAW_FILE_NAME":"VT_170706_040","RAW_FILE_NAME":"VT_170706_017"}
},
{
"Subject ID":"Study_Sample",
"Sample ID":"POTR_03_Plasma",
"Factors":{"Description":"POTR_03"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_043","RAW_FILE_NAME":"VT_170706_045","RAW_FILE_NAME":"VT_170706_047","RAW_FILE_NAME":"VT_170706_044","RAW_FILE_NAME":"VT_170706_046","RAW_FILE_NAME":"VT_170706_018"}
},
{
"Subject ID":"Study_Sample",
"Sample ID":"POTR_04_Plasma",
"Factors":{"Description":"POTR_04"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_049","RAW_FILE_NAME":"VT_170706_051","RAW_FILE_NAME":"VT_170706_053","RAW_FILE_NAME":"VT_170706_050","RAW_FILE_NAME":"VT_170706_052","RAW_FILE_NAME":"VT_170706_019"}
},
{
"Subject ID":"Study_Sample",
"Sample ID":"POTR_05_Plasma",
"Factors":{"Description":"POTR_05"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_055","RAW_FILE_NAME":"VT_170706_057","RAW_FILE_NAME":"VT_170706_059","RAW_FILE_NAME":"VT_170706_056","RAW_FILE_NAME":"VT_170706_058","RAW_FILE_NAME":"VT_170706_020"}
},
{
"Subject ID":"Study_Sample",
"Sample ID":"POTR_06_Plasma",
"Factors":{"Description":"POTR_06"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_061","RAW_FILE_NAME":"VT_170706_063","RAW_FILE_NAME":"VT_170706_065","RAW_FILE_NAME":"VT_170706_062","RAW_FILE_NAME":"VT_170706_064","RAW_FILE_NAME":"VT_170706_021"}
},
{
"Subject ID":"Study_Sample",
"Sample ID":"POTR_07_Plasma",
"Factors":{"Description":"POTR_07"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_067","RAW_FILE_NAME":"VT_170706_069","RAW_FILE_NAME":"VT_170706_071","RAW_FILE_NAME":"VT_170706_068","RAW_FILE_NAME":"VT_170706_070","RAW_FILE_NAME":"VT_170706_022"}
},
{
"Subject ID":"Study_Sample",
"Sample ID":"POTR_08_Plasma",
"Factors":{"Description":"POTR_08"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_073","RAW_FILE_NAME":"VT_170706_075","RAW_FILE_NAME":"VT_170706_077","RAW_FILE_NAME":"VT_170706_074","RAW_FILE_NAME":"VT_170706_076","RAW_FILE_NAME":"VT_170706_023"}
},
{
"Subject ID":"Study_Sample",
"Sample ID":"POTR_09_Plasma",
"Factors":{"Description":"POTR_09"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_079","RAW_FILE_NAME":"VT_170706_081","RAW_FILE_NAME":"VT_170706_083","RAW_FILE_NAME":"VT_170706_080","RAW_FILE_NAME":"VT_170706_082","RAW_FILE_NAME":"VT_170706_024"}
},
{
"Subject ID":"Study_Sample",
"Sample ID":"POTR_10_Plasma",
"Factors":{"Description":"POTR_10"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_085","RAW_FILE_NAME":"VT_170706_087","RAW_FILE_NAME":"VT_170706_089","RAW_FILE_NAME":"VT_170706_086","RAW_FILE_NAME":"VT_170706_088","RAW_FILE_NAME":"VT_170706_025"}
},
{
"Subject ID":"Study_Sample",
"Sample ID":"POTR_11_Plasma",
"Factors":{"Description":"POTR_11"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_091","RAW_FILE_NAME":"VT_170706_093","RAW_FILE_NAME":"VT_170706_095","RAW_FILE_NAME":"VT_170706_092","RAW_FILE_NAME":"VT_170706_094","RAW_FILE_NAME":"VT_170706_026"}
},
{
"Subject ID":"Study_Sample",
"Sample ID":"POTR_12_Plasma",
"Factors":{"Description":"POTR_12"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_097","RAW_FILE_NAME":"VT_170706_099","RAW_FILE_NAME":"VT_170706_101","RAW_FILE_NAME":"VT_170706_098","RAW_FILE_NAME":"VT_170706_100","RAW_FILE_NAME":"VT_170706_027"}
},
{
"Subject ID":"QC",
"Sample ID":"q3June2014_1c",
"Factors":{"Description":"Q-Standard plasma pool"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_103","RAW_FILE_NAME":"VT_170706_105","RAW_FILE_NAME":"VT_170706_107","RAW_FILE_NAME":"VT_170706_104","RAW_FILE_NAME":"VT_170706_106","RAW_FILE_NAME":"VT_170706_028"}
},
{
"Subject ID":"QC",
"Sample ID":"q3June2014_1d",
"Factors":{"Description":"Q-Standard plasma pool"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_109","RAW_FILE_NAME":"VT_170706_111","RAW_FILE_NAME":"VT_170706_113","RAW_FILE_NAME":"VT_170706_110","RAW_FILE_NAME":"VT_170706_112","RAW_FILE_NAME":"VT_170706_029"}
},
{
"Subject ID":"QC",
"Sample ID":"chearplasma_1c",
"Factors":{"Description":"CHEAR plasma pool"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_115","RAW_FILE_NAME":"VT_170706_117","RAW_FILE_NAME":"VT_170706_119","RAW_FILE_NAME":"VT_170706_116","RAW_FILE_NAME":"VT_170706_118","RAW_FILE_NAME":"VT_170706_030"}
},
{
"Subject ID":"QC",
"Sample ID":"chearplasma_1d",
"Factors":{"Description":"CHEAR plasma pool"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_121","RAW_FILE_NAME":"VT_170706_123","RAW_FILE_NAME":"VT_170706_125","RAW_FILE_NAME":"VT_170706_122","RAW_FILE_NAME":"VT_170706_124","RAW_FILE_NAME":"VT_170706_031"}
},
{
"Subject ID":"QC",
"Sample ID":"NIST_1958_2",
"Factors":{"Description":"NIST 1958"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"VT_170706_127","RAW_FILE_NAME":"VT_170706_129","RAW_FILE_NAME":"VT_170706_131","RAW_FILE_NAME":"VT_170706_128","RAW_FILE_NAME":"VT_170706_130","RAW_FILE_NAME":"VT_170706_032"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Eleven adolescents 12–20 years of age undergoing bariatric surgery at Cincinnati Children’s Hospital between 2006 and 2012 were offered enrollment in a prospective biospecimen repository protocol (Pediatric Obesity Tissue Repository [POTR]). Sample recruitment and other POTR features have been reported previously (Davidson et al. 2017). Intraoperatively, visceral adipose tissue (vAT) samples from the omentum, abdominal subcutaneous AT (sAT), and liver samples were obtained by the surgeon and processed immediately in an area adjacent to the operating room. All samples were snap-frozen in liquid nitrogen, then stored at −80°C. Plasma was collected pre-operatively after overnight fasting and stored at -80°C. Written informed consent was obtained from participants equal to or above 18 years old or from the parent or guardian if participants were less than 18 years old. The study was approved by the Institutional Review Board at Cincinnati Children’s Hospital.","SAMPLE_TYPE":"Blood (plasma)","STORAGE_CONDITIONS":"-80℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"The objective of the observational study was to evaluate the relationship between adipose and liver tissue POPs and the plasma metabolome. All participants underwent bariatric surgery at the time of tissue collection. No other treatment or intervention was evaluated."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Samples are prepared for metabolomics analysis using established methods (Johnson et al. (2010). Analyst; Go et al. (2015). Tox Sci). Prior to analysis, plasma aliquots were removed from storage at -80°C and thawed on ice. Each cryotube is then vortexed briefly to ensure homogeneity, and 50 μL transferred to a clean microfuge tube. Immediately after, the plasma is treated with 100 μL of ice-cold LC-MS grade acetonitrile (Sigma Aldrich) containing 2.5 μL of internal standard solution with eight stable isotopic chemicals selected to cover a range of chemical properties. Following addition of acetonitrile, plasma is then equilibrated for 30 min on ice, upon which precipitated proteins are removed by centrifuge (16.1 ×g at 4°C for 10 min). The resulting supernatant (100 μL) is removed, added to a low volume autosampler vial and maintained at 4°C until analysis (<22 h).","SAMPLEPREP_PROTOCOL_ID":"EmoryUniversity_HRM_SP_082016_01.pdf","SAMPLEPREP_PROTOCOL_FILENAME":"EmoryUniversity_HRM_SP_082016_01.pdf","PROCESSING_STORAGE_CONDITIONS":"Room temperature"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"The C18 column is operated parallel to the HILIC column for simultaneous analytical separation and column flushing through the use of a dual head HPLC pump equipped with 10-port and 6- port switching valves. During operation of the C18 method, the MS is operated in negative ion mode and 10 μL of sample is injected onto the C18 column while the HILIC column is flushing with wash solution. Flow rate is maintained at 0.4 mL/min until 1.5 min, increased to 0.5 mL/min at 2 min and held for 3 min. Solvent A is 100% LC-MS grade water, solvent B is 100% LC-MS grade acetonitrile and solvent C is 10mM ammonium acetate in LC-MS grade water. Initial mobile phase conditions are 60% A, 35% B, 5% C hold for 0.5 min, with linear gradient to 0% A, 95% B, 5% C at 1.5 min, hold for 3.5 min, resulting in a total analytical run time of 5 min. During the flushing phase (HILIC analytical separation), the C18 column is equilibrated with a wash solution of 0% A, 95% B, 5% C until 2.5 min, followed by an equilibration solution of 60% A, 35% B, 5% C for 2.5 min.","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Dionex Ultimate 3000","COLUMN_NAME":"Higgins endcapped C18 stainless steel (2.1mm x 50mm x 3µm)","FLOW_GRADIENT":"Flow rate is maintained at 0.4 mL/min until 1.5 min, increased to 0.5 mL/min at 2 min and held for 3 min","FLOW_RATE":"0.4- 0.5mL/min","COLUMN_TEMPERATURE":"60C","INTERNAL_STANDARD":"[13C6]-D-glucose, [15N,13C5]- L-methionine, [13C5]-L-glutamic acid, [15N]-L-tyrosine, [3,3-13C2]-cystine, [trimethyl- 13C3]-caffeine, [U-13C5, U-15N2]-L-glutamine","SAMPLE_INJECTION":"10 uL","ANALYTICAL_TIME":"5 min"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","LABORATORY_NAME":"Clinical Biomarkers Laboratory","OPERATOR_NAME":"Vilinh Tran","ACQUISITION_DATE":"July 2017","DATA_FORMAT":".RAW"},

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive HF hybrid Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"The high-resolution mass spectrometer was operated at 120,000 resolution and mass-to-charge ratio (m/z) range 85-1275. Probe temperature, capillary temperature, sweep gas and S-Lens RF levels were maintained at 200°C, 300°C, 1 arbitrary units (AU), and 45, respectively. Additional source settings were optimized for sensitivity using a standard mixture, tune settings for sheath gas, auxiliary gas, sweep gas and spray voltage setting were 45 AU, 25 AU and 3.5 kV, respectively. Maximum C-trap injection times were set at 100 milliseconds and automatic gain control target 1 × 106. During untargeted data acquisition, no exclusion or inclusion masses were selected, and data was acquired in MS1 mode only. Raw data files were then extracted using apLCMS (Yu et al. 2009) at five different peak detection settings that have been separately optimized for detection of a wide range of peak intensities and abundances. Peaks detected during each injection were aligned using a mass tolerance of 5 ppm (parts-per-million) and retention grouping was accomplished using non-parametric density estimation grouping, with a maximum retention time deviation of 30 seconds. The resulting feature tables were merged using xMSanalyzer, which identifies overlapping or unique features detected across the different peak detection parameters, and retains the peak with the lowest replicate CV and non-detects for inclusion in the final feature table (Uppal et al. 2013). All R-scripts for data extraction with apLCMS and data merging with xMSanalyzer are provided in the supplementary material. Uniquely detected ions consisted of m/z, retention time and ion abundance, referred to as m/z features. Prior to data analysis, triplicate m/z features averaged and filtered to remove those with triplicate coefficient of variation (CV) ≥ 100% and non-detected values greater than 10%.","CAPILLARY_TEMPERATURE":"300C","ION_SOURCE_TEMPERATURE":"200C","ION_SPRAY_VOLTAGE":"3.5kV","IONIZATION":"NEgative","MASS_ACCURACY":"5ppm","SOURCE_TEMPERATURE":"200C","MS_RESULTS_FILE":"ST001407_AN002351_Results.txt UNITS:Peak intensity Has m/z:Yes Has RT:Yes RT units:Seconds"}

}