Outcomes
anthro, FM
WEST, CLOT
The primary outcomes were height (cm) and knee-heel length (mm), and the primary effect estimate was difference in changes in height and in knee-heel length between the LNS interventions from baseline to week 12. Secondary outcomes included the following: weight (g), body composition by calibrated bioelectrical impedance analysis [FM, FFM, fat mass index (FMI), and fat-free mass index (FFMI) (kg/mHeight (or length for children below 2 years) was measured 3 times using a wooden board (Weigh and Measure, Maryland, United States of America), ensuring 5 points of contact with repositioning between measurements. Knee-heel length was measured using a digital caliper with a 0.01 mm resolution (Mitutoyo, Neuss, Germany) mounted with knee and heel caps cast in hard plastic. The distance between the knee (from the lateral condyle) and the heel (calcaneus) was measured 5 consecutive times on the left leg preferably as the child sat with both legs hanging on the edge of a table or on the caregiver’s lap.Participant and maternal weight were measured using an electronic double weighing scale (SECA 876, Hamburg, Germany) while maternal height was taken using a fixed wall stadiometer (SECA 206, Hamburg, Germany). MUAC and skinfolds were measured using a non-elastic MUAC tape (UNICEF SD, Copenhagen, Denmark) and a Harpenden skinfold caliper (Baty International, West Sussex, England), respectively. Skinfolds thickness were taken on the left side according to the caliper manufacturer’s instructions. For site referral and study inclusion, z-scores were calculated using the World Health Organization (WHO) field growth charts while the WHO anthro program (igrowup macro) was used to calculate z-scores in Stata 14 (Stata, College Station, Texas, USA). Body composition was measured in duplicate by bioelectrical impedance using Bodystat 500 machines at 50 kilohertz (KHz) (Bodystat, Isle of Man, United Kingdom). The mean impedance was converted to FFM using an equation developed for this specific population of 1- to 5-year-old Ugandan children with stunting [Sex was coded as female = 0 and male = 1, age was in months, height was in cm, and ZBlood samples were collected by venipuncture from the forearm in vacuum tubes. These were stored at ambient temperature, transported to a local lab, left to clot for 30 min, and centrifuged for 10 min at 3,500 rpm (2,300 g). Serum was then transferred to cryotubes and frozen at −20°C. Every week, samples were transported on ice to a biorepository and stored at −80°C (Integrated Biorepository of H3Africa Uganda, Makerere University). Samples were shipped on dry ice to UCPH for IGF-1 analysis and VitminLab (Willstaett, Germany) for α1-acid glycoprotein (AGP) analysis. Serum IGF-1 was analyzed on an Immulite 2000 Analyzer (Siemens Healthcare, GmbH) with intra- and inter-assay CVs of 1.9% to 4.2% and 4.2% to 7.2%, respectively. AGP was analyzed by sandwich enzyme-linked immunosorbent assay with assay variability between 5% and 14%.