A 7 ml blood sample was drawn from participants seated and rested by medical personnel using serum separation vacutainers according to the schedule above. Serum was allowed to clot for a minimum of 30 minutes but no longer than 60 minutes. After centrifugation serum was stored in duplicate plastic 1.5-ml Eppendorf tubes at -80 0 C. SHBG (nmol/L), LH (IU/L), FSH (IU/L) and TSH (mIU/L) were analysed on the Roche Modular System (Roche Diagnostics, Lewes, UK) by two-site sandwich immunoassay using electro chemiluminesence technology. Testosterone ( nmol/L), free T3 (FT3, pmol/L) and free thyroxine (FT4, pmol/L) were also analysed on the Roche Modular System but using competition immunoassay with albumin (g/L) measured using an endpoint colourimetric method (bromcreosol green). Testosterone and SHBG were used to calculate free testosterone (pmol/L) by the equation: {[(SHBG -T + 23)2 + (92 x T)] -(SHBG -T + 23)}/0.046 (Vermeulen et al. 1999 ). The somatotroph axis was assessed by assay of insulin-like growth factor-1 (IGF-1, nmol/L). Assay of IGF-1 was based on chemiluminescence technology using the IDS Isys instrumentation (calibrated to the new WHO international standard).
Modular system
Manufactured by Roche
Sourced in Switzerland, Germany, United Kingdom
The Modular system is a versatile laboratory equipment solution offered by Roche. Its core function is to provide a customizable and adaptable platform for various laboratory applications. The modular design allows for the integration of different components and modules to meet the specific needs of each laboratory.
Automatically generated - may contain errors
Lab products found in correlation
Delta plus xp, by Thermo Fisher Scientific (1 mentions)
Insulin, by Mercodia (1 mentions)
Nonesterified fatty acids, by ZenBio (1 mentions)
Cortisol, by R&D Systems (1 mentions)
Enzychrome triglyceride assay kit, by BioAssay Systems (1 mentions)
Rat insulin elisa kit, by Mercodia (1 mentions)
Rat adiponectin elisa kit, by Assaypro (1 mentions)
Rat leptin elisa kit, by BioVendor (1 mentions)
Solid phase 125i radioimmunoassay, by Siemens (1 mentions)
Elecsys probnp, by Roche (1 mentions)
Enzymatic kit, by Roche (1 mentions)
Modular analyser, by Roche (1 mentions)
Cobas e411 analyser, by Roche (1 mentions)
A0021, by Agilent Technologies (1 mentions)
Integra 800 system, by Roche (1 mentions)
Cobas 8000, by Roche (1 mentions)
917 analyzer, by Hitachi (1 mentions)
Dimension vista 1500 system, by Siemens (1 mentions)
Afinion as100, by Axis-Shield (1 mentions)
33 protocols using modular system
1
Serum Biomarker Profiling Protocol
2
Multimodal Steroid Profiling and Analysis
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Serum concentrations of adrenal and gonadal steroids were measured using liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, employing a validated multisteroid profiling method (24 ). In brief, serum steroids were extracted via liquid/liquid tert-butyl-methyl-ether (MTBE), evaporated, reconstituted, and analyzed by LC-MS/MS for cortisol and cortisone. Serum androgens and androgen precursors (DHEA, androstenedione, testosterone) were measured following oxime derivatization (25 (link), 26 (link)). Serum DHEA sulfate (DHEAS) was measured following protein precipitation (27 (link), 28 (link)). Steroid metabolite excretion analysis in 24-hour urine samples was carried out by gas chromatography-mass spectrometry (GC-MS) in selected-ion-monitoring (SIM) mode, as previously described (29 (link)).
Serum concentrations of sex hormone-binding globulin (SHBG) and luteinizing hormone (LH) were analyzed on the Roche Modular System (Roche Diagnostics, Lewes, UK) by 2-site sandwich immunoassay using electrochemiluminesence technology.
Serum concentrations of sex hormone-binding globulin (SHBG) and luteinizing hormone (LH) were analyzed on the Roche Modular System (Roche Diagnostics, Lewes, UK) by 2-site sandwich immunoassay using electrochemiluminesence technology.
3
Serum sFRP2 and Metabolic Markers
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Human serum sFRP2 levels were measured using a commercially-available ELISA (USCNK Life Science Inc, B2 Scientific Limited, UK). Intrassay and interassay coefficients of variation (CV) for this assay were < 10% and < 12% respectively. Insulin was measured using a commercially-available colorimetric ELISA (Mercodia, Uppsala, Sweden) with an in-house CV of < 5%. Glucose, triglyceride and HbA1c were analysed on a standard automated platform (Roche Modular system, Roche, Lewes, UK). Fasting glucose and insulin levels were used in the homeostasis model assessment computer model (HOMA2-IR) to generate estimates of insulin sensitivity. HOMA%B is considered to be a measure of pancreatic beta-cell function and is expressed as a percentage [20 (link)].
4
Metabolic Cage Urinalysis in Rats
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Rats were placed in metabolic cages for 14 hours on days 6 and day 13. Hematuria and leukocyturia were assessed by semiquantitative analysis of urinalysis reagent strips (Mutistix 8SG; Bayer, Pymble, Australia). Creatinine, protein, and albumin in serum were measured using an autoanalyzer (Roche Modular System; Hoffman-La Roche Ltd, Basel, Switzerland) at the Institute of Clinical Pathology and Medical Research, Westmead Hospital.
5
Metabolic Biomarker Assessment in Patients
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Serum specimens collected from the patient and control groups were stored at -80 °C until analysis. Routine biochemical tests were analyzed in the Central Biochemistry Laboratory, Cerrahpaşa Faculty of Medicine, İstanbul. Fasting blood glucose, total cholesterol, high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol and triglyceride concentrations were measured by enzymatic colorimetric methods, while aspartate aminotransferase and alanine aminotransferase (ALT) were assayed by kinetic ultraviolet methods on a Roche Modular System (Cobas, Roche GmbH, Germany). Insulin concentration was measured by solid phase sandwich enzyme-linked immunosorbent assay (DRG instruments GmbH, Germany). Insulin resistance was assessed using HOMA-IR, which was calculated using the following standard formula: glucose (mg/dL) x insulin (µIU/mL)/405 (18 ).
6
Standardized Biochemical Analysis Methods
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All biochemical parameters were determined by standard methods on a Roche-Modular System (Basel, Switzerland). In the multicenter study, all biochemical analyses were performed in a single center.25 (link)
7
Comprehensive Metabolic Profiling in Humans
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Liver biochemistry, electrolytes, urea, creatinine, cholesterol, triglycerides, and full blood counts were measured using standard laboratory methods (Roche modular system; Roche Ltd). Cortisol (R&D Systems), insulin (Mercodia), and nonesterified fatty acids (Zen-Bio) were measured using commercially available assays according to the manufacturers' instructions.
Microdialysate samples were collected in microvials and analyzed using a mobile photometric, enzyme-kinetic analyzer (CMA Iscus Flex) for glucose, pyruvate, lactate. and glycerol.
The enrichment of U-13C-glucose and D5-glycerol (1,1,2,3,3-D5) in plasma was determined by gas chromatography-mass spectrometry (model 5973; Agilent Technologies). Deuterium enrichment of the total body water pool was measured using the Gasbench II coupled online to a ThermoFinnigan Deltaplus XP isotope ratio mass spectrometer and the palmitate fraction of total plasma triglycerides using an automated gas chromatography high temperature conversion isotope ratio mass spectrometer ThermoFinnigan Delta Pus XP (ThermoFinnigan) (19 (link)).
Microdialysate samples were collected in microvials and analyzed using a mobile photometric, enzyme-kinetic analyzer (CMA Iscus Flex) for glucose, pyruvate, lactate. and glycerol.
The enrichment of U-13C-glucose and D5-glycerol (1,1,2,3,3-D5) in plasma was determined by gas chromatography-mass spectrometry (model 5973; Agilent Technologies). Deuterium enrichment of the total body water pool was measured using the Gasbench II coupled online to a ThermoFinnigan Deltaplus XP isotope ratio mass spectrometer and the palmitate fraction of total plasma triglycerides using an automated gas chromatography high temperature conversion isotope ratio mass spectrometer ThermoFinnigan Delta Pus XP (ThermoFinnigan) (19 (link)).
8
Thyroid Hormone Serum Analysis
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Blood samples were drawn from all study participants and serum was separated, aliquoted, and stored at −80 °C until analyses. Serum concentrations of TSH and free T4 were analyzed on the Roche Modular system (Roche Diagnostics Scandinavia AB, Solna, Sweden) as described previously [24 (link)]. The total CV of the TSH assay was ≤ 10% and the detection limit was 0.005 mIU/L. The total CV of the free T4 assay was ≤ 10% and the detection limit was 1.5 pmol/L.
9
Serum Enzyme and Lipid Analysis
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Serum AST and ALT activities were determined on a Roche Modular System (Roche Diagnostics) with a commercially available automated colorimetric system at the Institute of Clinical Chemistry, University Hospital Zurich, using a Hitachi P-Modul (Roche). Triglyceride concentrations in mouse liver homogenates were measured using EnzyChrome triglyceride assay kit (BioAssay Systems) as previously described by us20 (link).
10
Biochemical Profiling of Fasted Rats
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Biochemical parameters, insulin, leptin and adiponectin were analyzed at the termination of study. The 16-h-fasted rats were etherized by ethyl ether and sacrificed by cardiac exsanguination. Blood samples were placed in centrifuge tube for 30 min and then centrifuged at 3500g for 30 min at 4 °C to obtain serum. After laparotomy, internal organs (liver, kidney and pancreas), epididymal fat and perinephric fat were isolated and weighed. All isolated tissues and serum samples were stored at −80 °C before analysis. Biochemical parameters, such as fasted blood sugar (FBG), aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine, bilirubin, triglyceride (TG), total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) in serum, were measured using a Roche Modular system and all the kits were purchased from Cobas (Roche, Mannheim, Germany). The rat insulin ELISA kit (Mercodia AB), rat leptin ELISA kit (BioVendor) and rat adiponectin ELISA kit (Assaypro) were employed to measure the insulin, leptin and adiponectin, respectively.
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