Immulite 2000 xpi

Venous blood samples were drawn five times from the antecubital vein after an overnight fast between 06:30 and 07:30 to use immunoassay system for analyzing insulin‐like growth factor‐1 (IGF‐1), interleukin 6 (IL‐6) (Siemens Immulite 2000 XPI, Siemens Healthcare Diagnostics Products Ltd., Gwynedd, UK), tumor necrosis factor alpha (TNF‐α) (Siemens Immulite 1000, Siemens Healthcare Diagnostics Products Ltd., Gwynedd, UK), leptin (ELISA‐kit, BioVendor, Brno, Czech Republic/Dynex DS 2, Dynex Technologies, Chantilly), and photometric system for analyzing creatine kinase (CK) levels (Konelab 20 XTi). The sensitivity and interassay variance for these assays were 2.65 nmol/L and 7.6% for IGF‐1, 0.11 pg/mL and 17.6% for IL‐6, 0.19 pg/mL and 9.6% for TNF‐α and 3.2 U/L and 5.8% for CK. The samples were centrifuged (Megafire 1.0 R Heraeus, DJB Lab Care, Germany) at 2000 g for 10 min and frozen and transported to laboratories at the University of Jyväskylä for later analysis.

Plasma was precipitated with 125 µL of 10% sulfosalicylic acid (SSA), centrifuged, and the supernatant was used to determine EAA concentrations using the internal standard technique [28 (link)]. Phenylalanine and tyrosine enrichments were measured using the tert-butyldimethylsilyl derivative and gas chromatography-mass spectrometry. Ions of mass-to-charge ratio of 234, 235, and 239 for phenylalanine and of 466, 467, 468, and 470 for tyrosine were monitored with electron impact ionization and selective ion monitoring. Serum insulin concentrations were measured using a Siemens Immulite 2000 XPi (Siemens Medical Solutions USA, Inc., Malvern, PA, USA).
Muscle samples were weighed, and tissue proteins were precipitated with 0.5 mL of 4% SSA. Samples were then homogenized, centrifuged, and the muscle pellet (bound protein) was washed, dried, and hydrolyzed in 0.5 mL of 6 N HCl at 105 °C for 24-h. Mixed muscle-bound protein enrichments were determined as described above for plasma enrichments.

BTMs and sclerostin were measured among a subgroup of participants with T1D (n = 61) and controls (n = 57) who had their blood sampled in the morning before 10:00 am after a minimum 10-h fast. Serum samples were frozen at −80 °C until batch analysis at the study end. Serum CTX [CV <4%], P1NP [CV <2%], and N-MID osteocalcin [CV <2%] were measured by automated electrochemiluminescence immunoassay (Cobas e411, Roche Diagnostics). These measurements were performed at the Centre Hospitalier de l’Université de Montréal, a clinical laboratory with extensive experience in the measurement of BTMs. Serum sclerostin [CV 5%] was measured by ELISA using TECO kit, as per manufacturer’s protocol (Sclerostin TECO High Sensitive; TECOmedical Group).
All participants had PTH (Siemens Immulite 1000, Siemens Healthineers), 25OHD (Siemens Advia Centaur XPT, Siemens Healthineers; interassay CV 8%), and IGF-1 (Siemens Immulite 2000 xPi, Siemens Healthineers) measured by automated chemiluminescent immunoassay. Other biochemical tests were performed using standard automated techniques: HbA_1C, hemoglobin, creatinine, lipid profile, thyrotropin (TSH), serum total calcium, albumin, phosphate, anti-transglutaminase antibodies and immunoglobulin A, and spot urine albumin-to-creatinine ratio. eGFR was calculated using the Chronic Kidney Disease Epidemiologic Collaboration equation.²⁹ (link)

Venous blood samples were drawn three times into Vacutainer® gel tube from the antecubital vein after an overnight fast between 06:30 and 07:30 to analyze serum concentrations of testosterone (TES), cortisol (COR), sex hormone binding globulin (SHBG), IGF‐1, and insulin‐like growth factor binding protein‐3 (IGFBP‐3) (Siemens Immulite 2000 XPI, Siemens Healthcare, USA). The sensitivity and interassay coefficients of variance for these assays were 0.5 nmol/l and 8.2% for TES, 5.5 nmol/l and 7.9% for COR, 0.02 nmol/l and 5.2% for SHBG, 2.6 nmol/l and 7.1% for IGF‐1, and 0.3 nmol/l and 8.4% for IGFBP‐3. The samples were centrifuged (Megafire 1.0 R Heraeus, DJB Lab Care, Germany) after 30 min at 2000 g for 10 min, frozen, and transported to the laboratory for later analysis.

After the venous blood samples were obtained from the antecubital vein, the samples were centrifuged at 2000 g for 20 min at room temperature for serum separation. Serum samples were then frozen and stored at −20°C for later analysis. The measurements of serum 25(OH)D were performed using electrochemiluminescence immunoassays (ECLIA). Levels of TES and SHBG were measured by electrochemiluminescence immunoassay (Immulite 100; Siemens Healthcare Diagnostics Products Ltd., Gwynedd, United Kingdom). IGF-1 was analyzed by electrochemiluminescence immunoassay (Siemens Immulite 2000 XPI, Siemens Healthcare Diagnostics Products Ltd., Gwynedd, United Kingdom). The inter-assay coefficients of variance (CV) for assays of TES, SHBG, and IGF-I were 7.0%–7.2%, 4.5%–6.2%, and 3.7%–7.4%, and that of sensitivity 0.5 nmol/L, 0.02 nmol/L, and 2.6 nmol/L, respectively.

Venous blood samples were drawn from the antecubital vein after an overnight fast 1 day prior to the field training. Serum concentrations of testosterone (TES), cortisol (COR), sex hormone binding globulin (SHBG), insulin-like growth factor-1 (IGF-1) and dehydroepiandrosterone (DHEA) and C-reactive protein (CRP) were analyzed (Siemens Immulite 2000 XPI, Siemens Healthcare, Malvern, PA, USA). The sensitivity and interassay coefficients of variance for these assays were 0.5 nmol/L and 7.8% for TES, 5.5 nmol/L and 6.5% for COR, 0.02 nmol/L and 5.7% for SHBG, 2.6 nmol/L and 7.8% for IGF-1, 0.08 µmol/L and 7.6% for DHEA and 0.1 mg/L and 7.3% for CRP. The samples were centrifuged (Megafire 1.0 R Heraeus, DJB Labcare Ltd., Buckinghamshire, UK) after 30 min at 2000× g for 10 min, frozen, and transported to the laboratory for later analysis.