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Total circulating levels of adiponectin vs transcript abundance in omental fat following the MIQE guidelines in a group of morbidly obese patients
Author(s) -
Mobula Ambie Zanga,
Alley Joshua,
Samy Sanjay,
Fitzgerald Laura,
Ariza Magnolia
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.1153.9
Subject(s) - adiponectin , medicine , adipose tissue , obesity , endocrinology , biology , insulin resistance
Obesity has a major role to play in multiple metabolic disorders including high blood pressure, type 2 diabetes and even cancer. Adiponectin is a peptide hormone expressed in omental adipose tissues. The goal of this study is to measure how adiponectin transcript abundance in omental fat compares to total circulating levels in circulation. The assays are performed in a group of patients presented with extreme excess weight that correspond to IRB approved clinical trial GHS # 1207–27. The expression of adiponectin is assessed following the minimum information for publication of quantitative real‐time PCR experiments (MIQE) guidelines which outline the minimum information that is essential in the evaluation of qPCR assays, increasing reliability, transparency and integrity of the experiments. Adiponectin transcript abundance is measured in RNA extracted from omental fat, followed by first strand cDNA synthesis using both oligodT and random primers. Target adiponectin and reference beta‐hemoglobin and/or GAPDH transcripts are amplified using SYBR Green. Total serum adiponectin is quantified using enzyme‐linked immunosorbent commercial assay. Both expressed and circulating levels of adiponectin are compared to metabolic clinical parameters. Funding Cornell University Center on the Microenvironment and Metastasis (CMM) outreach pilot grant and Guthrie Foundation Investigator‐Initiated Research Grant.