101.2 ± 1.9 years and 112 women, mean age 100.8 ±
1.7 years). The group of semi-supercentenarians (i.e. persons who reach the age of 105 years, 105+)
consisted of 70 subjects (16 men, mean age 105.6 ± 0.9 years and 54 women, mean age 106.3 ± 1.6
years). The group of centenarian offspring (CENTOFF) consisted of 308 subjects (125 men, mean
age 70.2 ± 6.5 years and 183 women, mean age 70.8 ± 7.1 years), having a centenarian or semisupercentenarian parent born between 1899 and 1909. The group of controls (CTRL) consisted of
150 age-matched subjects (80 men, mean age 70.5 ± 6.7 years and 70 women, mean age 70.3 ± 6.5
years) with no long-lived parents. The lists of centenarians, semi-supercentenarians and their
offspring were obtained by the Register Office, while the controls were identified by checking the
birth and death dates of their parents in paper population records from Registers Office. All
participants signed the informed consent before undergoing the questionnaires, measurements and
blood sampling. The study protocol was approved by the Ethical Committee of Sant’OrsolaMalpighi University Hospital (Bologna, Italy).
Measurement tools. Functional status was assessed by ADL-Activities of Daily Living scale
(scores ranging from 0 [all functions lost] to 5 [all functions preserved]) (20).
Physical performance was assessed by Handgrip strength test measuring the maximum
isometric strength of the hand and forearm muscles. Handgrip strength was measured using a handheld dynamometer (SMEDLYS’ dynamometer, Scandidact, Kvistgaard, Denmark) for two
performances with each hand. The best performance was selected for the analysis. Cognitive status
was assessed by SMMSE-Standardized Mini-Mental State Examination test (21). Depression status
was investigated by Geriatric Depression Scale short form (GDS, 15 items) (22).
History of past and current diseases was accurately collected by checking the participants'
medical documentation and addressing the major age-related pathologies. Current use of medication
(including inspection of the drugs by the interviewer) was recorded. Thyroid dysfunctions
suggestive of clinical hypothyroidism and hyperthyroidism, subclinical hypothyroidism and
by Bukkyo University Library user
on 31 August 2018
Accepted Manuscript
6
hyperthyroidism, central hypothyroidism and NTIS were identified based on serum FT4, FT3 and
TSH concentrations, as indicated in Supplemental Table 1. The criteria for the classification of the
above-mentioned conditions were defined with respect to population-based reference ranges of
different study centers.
Laboratory measurements. Overnight fasting blood samples were obtained in the morning.
Serum was obtained after clotting and centrifugation at 760 g for 20 min at 4°C, rapidly frozen and
stored at -80°C. Plasma was obtained within 2 hours from venipuncture by centrifugation at 2000 g
for 20 min at 4°C, rapidly frozen and stored at -80°C. Serum total and HDL cholesterol,
triglycerides, C-Reactive Protein (CRP) and glycaemia were measured by standard biochemical
assays. Thyroid hormones (FT3, FT4) and TSH were measured according to standard procedures in
three study centers (Bologna: ElectroChemiLuminescence ImmunoAssay (ECLIA), Elecsys Roche
Diagnostics S.p.A.; Milan: ElectroChemiLuminescence ImmunoAssay (ECLIA), Cobas Roche
Diagnostics GmbH; Cosenza: ChemiLuminescence ImmunoAssay, ADVIA Centaur®, Siemens
Healthcare Diagnostics Inc.). Each study center used its appropriate population-based reference
ranges to define euthyroidism and thyroid dysfunctions/pathologies as shown in Supplemental
Table 1. Serum insulin was measured by a chemiluminescent immunoassay (LIAISON® Insulin
assay, DiaSorin, Saluggia, Italy) and analyzed by the LIAISON® Analyzer. Insulin resistance status
was assessed as homeostasis model assessment of insulin resistance (HOMA-IR) according to the
following formula [67]: insulin (IU/mL) x glucose (mmol/L) /22.5 (23).
Hormones Mesurement