This study is a secondary analysis of a randomized, parallel-controlled, double-blinded, 12-wk diet intervention study that was designed to examine the effects of milk fat, with a high or low MC-SFA content, and of milk protein, whey or casein, on post-prandial lipemia. Design and methods of the original study were described in detail previously (Bohl et al. 2015, doi: 10.3945/ajcn.114.097923). Briefly, 52 participants were randomized in a 2 × 2 factorial design to one of 4 diets: high MC-SFA + casein, low MC-SFA + casein, high MC-SFA + whey, and low MC-SFA + whey. In this paper, we examine the effects of MC-SFAs on whole genome gene expression in subcutaneous adipose tissue in a subpopulation of 12 subjects, 6 in the high MC-SFA + casein group and 6 in the low MC-SFA + casein groups.
Growth protocol
Participants needed to be ≥18 y of age, have an abdominal circumference of ≥80 cm for women or ≥94 cm for men, and be weight stabile for at least 3 months prior to inclusion. Furthermore, they could not be diabetic, pregnant, lactating, or suffer from severe cardiovascular, renal, endocrine, or psychiatric disease. All participants gave written informed consent to participate in the study and the study protocol was approved by the Central Denmark Region Committees on Health Research Ethics. The study was registered at clinicaltrials.gov as NCT01472666.
Extracted molecule
total RNA
Extraction protocol
Total RNA was isolated using RNeasy micro kit from Qiagen. RNA integrity was checked on chip analysis (Agilent 2100 bioanalyzer, Agilent Technologies, Amsterdam, the Netherlands) according to the manufacturer's instructions. RNA was judged as suitable for array hybridization only if samples exhibited intact bands corresponding to the 18S and 28S ribosomal RNA subunits, and displayed no chromosomal peaks or RNA degradation products (RNA Integrity Number > 7.0).
Label
biotin
Label protocol
One hundred nanogram of RNA was used for whole transcript cDNA synthesis with the Ambion WT expression kit [catalog number 4411974] (Applied Biosystems/Life Technologies, Nieuwekerk a/d IJssel, The Netherlands).
Hybridization protocol
Hybridization and washing of the Affymetrix GeneChip Human Gene 1.1 ST arrays were performed on a GeneTitan Instrument (Affymetrix, Santa Clara, CA) according to the manufacturer's recommendations.
Scan protocol
Arrays were scanned on an Affymetrix GeneTitan instrument (Affymetrix, Santa Clara, CA).
Description
G144_A07_22_m2t0_mn3
Data processing
Expression estimates were calculated applying the RMA algorithm in the Bioconductor library 'Oligo' (v1.34.2).
Dietary medium-chain saturated fatty acids induce gene expression of energy metabolism-related pathways in adipose tissue of abdominally obese subjects
