Serum gel monovette

Venous blood sampling was performed using ethylenediaminetetraacetic acid (EDTA) and serum-gel monovettes (Sarstedt, Germany). Subsequently, the following parameters were analyzed in the central laboratory of the BG University Hospital Bergmannsheil Bochum immediately after sampling: glycated hemoglobin (HbA_1c), total cholesterol, HDL-cholesterol, LDL-cholesterol, and thyroid-stimulating hormone (TSH). Conversely, Hcy was analyzed at the Immunological Laboratory of the University Hospital of Pediatrics and Adolescent Medicine, St. Josef-Hospital, Bochum, where the procedure has been established. Thus, within a time lag of 1 week to the first appointment, fasting venous blood samples were collected in EDTA monovettes (Sarstedt, Germany) and were promptly put on ice. Plasma was obtained by centrifugation (4000x g, 10 min, 4 °C) and stored at − 80 °C until further analysis of Hcy. Upon completion of the entire sample collection, an Enzyme-linked Immunosorbent Assay (ELISA) Kit (EKX-UAP3O8–96, Nordic BioSite, Sweden) was used to measure homocysteine levels in plasma in a total of three batches.

Fifteen male healthy participants, age = 27.5 ± 3.1 years, weight = 78 ± 7.7 kg, height = 1.81 ± 0.05 m, BMI = 23.9 ± 1.9 kg/m² were recruited by German Sport University Cologne, Cologne, Germany to study the effect of exercise on the serum asprosin level (Supplementary Table S7). Inclusion criteria were age from 20 to 35 years and physical fitness. Exclusion criteria were musculoskeletal disorder, acute and chronic injuries, trauma or surgery of lower extremities’ joints. Subjects were asked not to exercise 24 h prior to the study and to come to the institute either by car or public transportation. The subjects remained seated for 30 min before the first blood sampling (t0). Further blood samples were drawn immediately afterwards (t1), and then 30 min (t2), 60 min (t3), and 120 min (t4) after running. Blood (5.0–7.5 ml) was collected by venipuncture using serum-gel monovettes1 (Sarstedt, Nümbrecht, Germany). After blood coagulation (30 min, RT), serum was isolated by centrifugation (10 min, 3000 rpm) and stored at − 80 °C.