Association of Serum Fibroblast Growth Factor 21 (FGF-21) Levels With Resting Metabolic Rate (RMR), in Children and Adolescents With Hashimoto's Thyroiditis.
Conditions: Hashimoto Disease
Sponsor: Aristotle University Of Thessaloniki

Official Title: Association of Serum Fibroblast Growth Factor 21 (FGF-21) 
Levels With Resting Metabolic Rate (RMR), in Children and 
Adolescents With Hashimoto's Thyroiditis 
 
NCT number: 02725879 
 
 
Date: 01-10-2015 
 
 
Study protocol: 
 
Participants 
Children and adolescents, aged 5–18 years, will be screened for Hashimoto’s 
thyroiditis/ chronic autoimmune thyroiditis (AIT) at the Pediatric Endocrinology Outpatient 
Clinic of Papageorgiou General Hospital and AHEPA University Hospital of Thessaloniki, 
Greece. Diagnosis of AIT will be based on the presence of anti-thyroid autoantibodies 
(Anti-TPOAb and/or Anti-TgAb) and one or more of the following: clinical symptoms of 
thyroid dysfunction, goiter, or diffuse/irregular hypoechogenicity of the thyroid gland 
during ultrasound examination [1]. The first 30 patients with subclinical hypothyroidism 
who will receive standard levothyroxine (LT4) treatment [2], will comprise the “AIT 
subclinical hypothyroid group”. Another 30 participants with AIT and euthyroidism at the 
time of enrolment will comprise the “AIT euthyroid group”, whereas 30 age- and sex-
matched healthy subjects will be also enrolled as “Control group” in the study. 
All participants will be present with a normal body mass index (BMI) for their age and 
sex, were drug-naive for at least 3 months, followed no special diet, and did not present any 
chronic and/or acute disease or menstrual disorder. The the “AIT subclinical hypothyroid 
group” will be re-assessed six months after starting LT4 treatment. 
Clinical and Biochemical Data 
Height will be measured to the nearest millimeter with a wall-mounted stadiometer 
(Harpenden Stadiometer, Holtain Limited, Crosswell Wales, U.K.). Waist, hip, and mid-
upper arm circumference (MUAC) will be measured with a Seca 201 measuring tape 
(Hamburg, Germany), and body weight will be assessed with a Seca 711 scale (Hamburg, 
Germany). Body fat (BF) will be assessed by the same experienced investigator using a 
skinfold caliper (Harpenden Skinfold Caliper, Baty International, West Sussex, U.K.) and 
the equations proposed by Slaughter et al. [3]. Fat mass (FM), fat-free mass (FFM), FM 
index (FMI), and FFM index (FFMI) will be calculated [4,5]. Body mass index (BMI) will 
be calculated, and standard deviation scores (SDS) for BMI, height and skinfolds will be 
determined from the WHO growth charts using the LMS growth software. All subjects will 
undergo a complete physical examination, including posterior palpation of the thyroid 
gland, and will be classified according to their puberty, applying Marshall and Tanner 
criteria. 
Resting Metabolic Rate (RMR) will be measured after a 12 h fast with a portable 
indirect calorimeter (FitMateTM, Cosmed, Rome, Italy) [6], using a pediatric face mask 
following the protocols proposed by the study conducted by Fullmer et al. [7]. 
Blood samples will be collected after overnight fasting, and serum levels of 
biochemical parameters were measured using standard methods and an ARCHITECTc 
16000 clinical chemistry system (Abbott, Abbott Park, IL, USA). Concentrations of insulin, 

thyroid-stimulating hormone (TSH), free triiodothyronine (FT3), free thyroxin (FT4), as 
well as anti-thyroid peroxidase antibody (Anti-TPOAb), and thyroglobulin antibody (Anti-
TgAb) titers were measured with an ADVIA Centaur XPT Immunoassay System (Siemens 
Healthcare GmbH, Erlangen, Germany). Laboratory’s reference range for TSH, FT4, and 
FT3 levels are 0.80–3.99 μIU/L, 10.55–20.72 pmol/L, and 4.21–7.57 pmol/L, respectively. 
The positive cut-off value of Anti-TPOAb and Anti-TgAb titers is>60 IU/mL. A thyroid 
gland ultrasound will be performed by the same radiologist at the beginning of the study. 
Serum FGF-21 levels will be measured in patients with subclinical hypothyroidism and 
the control group. FGF-21 levels were determined in pg/mL using the Solid Phase 
Sandwich ELISA method according to the manufacturer’s protocol (Quantikine® Elisa, 
Human FGF-21 immunoassay DF 2100, R&D Systems Europe Ltd., Abingdon Science 
Park, Abingdon, U.K.) with a sensitivity of 8.69 pg/mL, intra-assay CV < 4%, inter-assay 
CV < 5% and an assay range of 31.3–2,000 pg/mL. 
In order we have indirect information regarding the dietary state of participants, all 
participants, with the help of their parents and/or caregivers, will complete the KIDMED 
questionnaire at their first visit. KIDMED questionnaire consists of 16 diet-related 
questions. A total score of 0–3 reflects a poor adherence to the Mediterranean diet, 4–7 an 
average compliance, and a score of 8–12 a suitable adherence [8,9]. 
 
[1] 
Caturegli P, De Remigis A, Rose NR. Hashimoto thyroiditis: Clinical and 
diagnostic criteria. Autoimmun Rev 2014;13:391–7. 
https://doi.org/10.1016/j.autrev.2014.01.007. 
[2] 
Vigone MC, Capalbo D, Weber G, Salerno M. Mild Hypothyroidism in 
Childhood: Who, When, and How Should Be Treated? J Endocr Soc 
2018;2:1024–39. https://doi.org/10.1210/js.2017-00471. 
[3] 
Slaughter MH, Lohman TG, Boileau RA, Horswill CA, Stillman RJ, van Loan 
MD  et al. Skinfold equations for estimation of body fatness in children and 
youths. Hum Biol 1988;60:709–23. 
[4] 
Al Shammari E, Bano R, Suneetha E, Alshammri ARH. FFM Index, FM Index 
and PBF in Subjects with Normal, Overweight, and Obese BMI in Saudi 
Arabia Female Population. J Food Res 2015;5:40. 
https://doi.org/10.5539/jfr.v5n1p40. 
[5] 
Wendel D, Weber D, Leonard MB, Magge SN, Kelly A, Stallings VA, et al. 
Body composition estimation using skinfolds in children with and without 
health conditions affecting growth and body composition. Ann Hum Biol 
2017;44:108–20. https://doi.org/10.3109/03014460.2016.1168867. 
[6] 
Nieman DC, Austin MD, Benezra L, Pearce S, McInnis T, Unick J, et al. 
Validation of cosmed’s FitMateTM in measuring oxygen consumption and 
estimating resting metabolic rate. Res Sport Med 2006;14:89–96. 
https://doi.org/10.1080/15438620600651512. 
[7] 
Fullmer S, Benson-Davies S, Earthman CP, Frankenfield DC, Gradwell E, Lee 
PSP, et al. Evidence Analysis Library Review of Best Practices for Performing 
Indirect Calorimetry in Healthy and Non-Critically Ill Individuals. J Acad Nutr 
Diet 2015;115:1417-1446.e2. https://doi.org/10.1016/j.jand.2015.04.003. 
[8] 
Serra-Majem L, Ribas L, Ngo J, Ortega RM, García A, Pérez-Rodrigo C, et al. 

Food, youth and the Mediterranean diet in Spain. Development of KIDMED, 
Mediterranean Diet Quality Index in children and adolescents. Public Health 
Nutr 2004;7:931–5. https://doi.org/10.1079/phn2004556. 
[9] 
Štefan L, Prosoli R, Juranko D, Čule M, Milinović I, Novak D, et al. The 
reliability of the mediterranean diet quality index (KIDMED) questionnaire. 
Nutrients 2017;9. https://doi.org/10.3390/nu9040419. 
 
 
 

Official Title: Association of Serum Fibroblast Growth Factor 21 (FGF-21) 
Levels With Resting Metabolic Rate (RMR), in Children and 
Adolescents With Hashimoto's Thyroiditis 
 
NCT number: 02725879 
 
 
Date: 01-10-2015 
 
 
Statistical Analysis 
 
Statistical analysis will be performed using IBM SPSS Statistics (SPSS Inc., Chicago, 
IL, USA). Continuous variables will be tested for normal distribution by the Kolmogorov–
Smirnov or Shapiro–Wilk test. Data will be presented as mean ± standard deviation (SD) or 
medians with lower or upper quartiles. The differences between the examined groups of 
individuals will be investigated using the Kruskal–Wallis test and ANOVA within the GLM 
function with Box-Cox transformation of the response variable to improve normality. The 
differences between the two groups of individuals will be assessed using the Tukey post-
hoc and Fisher least significant difference tests. When data are paired (e.g., before vs. after), 
we will account for subject ID in our model. The correlations between two continuous 
variables will be investigated using the Spearman rank correlation test. The level of 
statistical significance is set at p < 0.05. 
 
 
 
 
 
 
 
 
 
 
 
 
 

Official Title: Association of Serum Fibroblast Growth Factor 21 (FGF-21) 
Levels With Resting Metabolic Rate (RMR), in Children and 
Adolescents With Hashimoto's Thyroiditis 
 
NCT number: 02725879 
 
Date: 01-10-2015