SARAS KIDS children India
PMMST children The Gambia
Weight, standing and sitting height; mid-upper-arm circumference; head, chest and abdominal circumferences, skinfolds (triceps, biceps, sub-scapular and supra-iliac) using standardised protocols.
After 5 min seated at rest. Mean of 3 readings of systolic and diastolic pressure from left upper arm. Instrument: OMRON HEM7080.
Plasma glucose concentrations after an overnight fast of ≥8 h and 30 and 120 min after a 1.75 g/kg oral anhydrous glucose load. Measured by standard enzymic methods on an autoanalyzer (India: Hitachi 902, Roche Diagnostics, Mannheim, Germany; The Gambia: Cobas Integra 400 Plus Biochemistry Analyzer, Roche Diagnostics).
Plasma insulin fasting and 30 mins after the glucose load Measured by a Mercodia ELISA assay on a Victor 2 analyzer, Turku, Finland, in India and by an SM-chemiluminescence method on an Architect i1000 Plus analyzer, Abbott in The Gambia.
Plasma fasting total, LDL- and HDL-cholesterol and triglycerides by standard enzymic methods (India: Hitachi 902; The Gambia: Cobas Integra 400 Plus).
Total and regional (arms, legs, trunk, android and gynoid) fat mass, lean mass and body fat % using dual-energy x-ray absorptiometry (DXA, Lunar Prodigy in India and Lunar iDXA in The Gambia, GE Medical Systems, GE Lunar Corporation, Madison USA).
Bone area (BA), bone mineral content (BMC), and bone mineral density (BMD) measured using dual-energy x-ray absorptiometry (DXA; Lunar Prodigy in India and Lunar iDXA in The Gambia).
Tibial total and trabecular volumetric bone mineral density (vBMD), and BA; and diaphysial BA, cortical area, thickness, BMC, cortical vBMD and strength (cross-sectional moment of inertia) measured using peripheral quantitative computed tomography (pQCT; Stratec XCT 2000, Stratec Ltd., Pforzheim, Germany).
Three core tests from the Kaufman Assessment Battery for children, 2nd edition, 2004 (KABC II) – Atlantis (learning ability, long-term storage and retrieval, associative memory); Word order (memory span, short-term memory, working memory); Pattern reasoning (reasoning ability, induction, deduction, fluid reasoning) .
Additional tests from the Wechsler Intelligence Scale for Children (WISC): Kohs block design (visuo-spatial problem-solving, visual perception and organisation); Coding-Wisc III (visual-motor processing speed and co-ordination, short-term memory, visual perception, visual scanning, cognitive flexibility, attention); Verbal fluency (a) animals, (b) names (broad retrieval ability, speed and flexibility of verbal thought processes, neuropsychological test of language production) [39,40,41].
DNA and RNA
Whole blood collected into EDTA tubes and DNA isolated using Qiagen DNA Blood Midi Kit. DNA methylation measured in a single laboratory (CSIR-Centre for Cellular and Molecular Biology, Hyderabad, India) using (genome wide) Illumina Infinium MethylationEPIC arrays and (locus-specific) bisulfite sequencing on Pyromark96 (see main text for more details). High-resolution genotype data generated using Illumina Global Screening Array. Buccal DNA obtained using Isohelix buccal swabs in India and Mawi iSwab kits in The Gambia. Whole blood samples collected into Paxgene tubes for later RNA isolation.
Full blood count
Hemoglobin, red cell count and indices, differential white blood cell count (India: Pentra XL Retic analyzer, Horiba Medical, Montpellier, France; The Gambia: Medonic hematology analyzer, Spanga, Sweden).