Blood samples were taken in the morning, in a fasting state. Blood was collected into siliconized vacuum tubes containing either K2 EDTA (Becton-Dickinson, Oxford, UK) for blood count analysis (Advia 2120, Siemens, Germany) or lithium heparin for clinical chemistry testing (Modular Analytics, Roche, CH). Clinical chemistry and immunochemistry tests were performed on Cobas® 6000 c501 and e601 module (Roche Diagnostics GmbH, Penzberg, Germany), according to the manufacturer's specifications and using proprietary reagents. The panel of tests included the following: fasting glucose concentration (GLU), total cholesterol (CHOL), HDL cholesterol (HDL), LDL cholesterol (LDL), triglycerides (TRY). In addition, also c-reactive protein (CRP) and haemoglobyn (Hb) were obtained by standard methods. The instrument was calibrated against appropriate proprietary reference standard materials and verified with the use of proprietary quality controls. Our evaluation of the within-run precision by internal quality control on the Cobas® 6000 c501 and e601module (Roche Diagnostics GmbH) showed low coefficients of variation.
E601 module
Manufactured by Roche
Sourced in Germany, United Kingdom
The E601 module is a component of Roche's laboratory equipment. It serves as a core processing unit, responsible for performing various analytical tasks. The module's primary function is to facilitate data processing and management within the larger laboratory system.
Automatically generated - may contain errors
Lab products found in correlation
Cobas 6000, by Roche (5 mentions)
Electrochemical luminescence analyzer, by Roche (2 mentions)
Cobas 6000 c501, by Roche (1 mentions)
K2edta, by BD (1 mentions)
Advia 2120, by Siemens (1 mentions)
Cobas integra 400 plus, by Roche (1 mentions)
Kx 21n, by Sysmex (1 mentions)
D10 set, by Bio-Rad (1 mentions)
Elecsys amh immunoassay, by Roche (1 mentions)
Cobas 6000 system, by Roche (1 mentions)
Elecsys estradiol 3, by Roche (1 mentions)
9 protocols using e601 module
1
Fasting Blood Sampling and Analysis
2
Hormonal Response to Exercise
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Blood samples were collected 07:00–09:00 h, 48–72 h following the last exercise session as previously described (21 (link)). Serum concentrations of TT, SHBG and cortisol were measured by electrochemiluminescent immunoassay on the E601 module of the Roche Cobas 6000 (Burgess Hill, West Sussex, UK). Inter-assay coefficients of variation (CV) over a six-month period were 4.5, 2.4 and 4.2% for TT, SHBG and cortisol, respectively. Free-T was calculated using the Vermueulen equation (22 (link)).
3
Assessing Hormonal Changes in Exercise
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Blood samples from each participant were collected at phase A, B and C, at 07:00–09:00 h, 48–72 h following the last exercise session as previously described (23 (link), 24 (link), 25 (link), 26 (link)). Serum concentrations of TT, sex hormone-binding globulin (SHBG) and cortisol were determined by electrochemiluminescent immunoassay on the E601 module of the Roche Cobas 6000. Inter-assay CVs over a six-month period were 4.5, 2.4 and 4.2% for TT, SHBG and cortisol respectively. Analyses were carried out in a clinical pathology laboratory (Royal Glamorgan Hospital, Wales, UK). Free-T was calculated using the Vermeulen formula (27 (link)), which has been validated against equilibrium dialysis (28 (link)). The testosterone:cortisol ratio (T:C) was calculated by the following equation: T:C = 100·(TT ÷ cortisol).
4
Comprehensive Blood Biomarker Assessment
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Analysis: a complete blood count (CBC) was performed by a full automated hematology analyzer (Sysmex, KX21N, Kobe, Japan). Using a chemistry autoanalyzer device (Cobas Integra 400 Plus, Roche Diagnostics, Germany), following the manufacturer’s instructions, routine biochemical analysis of serum albumin, aspartate aminotransferase (AST), alanine aminotransferase (ALT), bilirubin (total and direct), alkaline phosphatase (ALP), gamma GT (GGT), total cholesterol (TC), triacylglycerol (TAG), and high-density lipoprotein-cholesterol (HDL-C) was performed. To measure plasma AFP, electro-chemiluminescence immunoassay (ECLIA) using a Cobas 6000, e601 module (Roche Diagnostics, Germany) was used. Finally, blood insulin was measured using the enzyme immunoassay (Hyperion Inc, Miami, FL).
5
Quantification of AFP-L3 Biomarker
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Blood samples were collected when diagnosed and before any interventions, then were frozen at -80°C before use. The levels of AFP and AFP-L3 were detected using AFP testing reagents supplied by Roche (Germany) through an electrochemical luminescence analyzer (e601 module, Roche, Germany). Prior to AFP-L3 detection, the blood samples need to be pre-processed. Every AFP-L3 portion supernatant was split from the blood samples via a Hotgen Biotech glycosyl capture spin-column. The primary mechanism of the spin-column was that the spin-column first pre-loaded Lens culinaris agglutinin to have the fucosylated stuff thoroughly captured, followed by a simple sugar added to compete with the aforementioned captured fucosylated stuff to bind to Lens culinaris agglutinin. Hence, the fucosylated stuff in the sera, the AFP-L3 included, was consequently eluted from the spin-column. After cleaning, free AFP was removed, and AFP-L3 was obtained by elution and centrifugation. Then the Roche electrochemical luminescence analyzer was applied to detecting the contents of total AFP and AFP-L3 respectively. the AFP-L3 percentages were calculated accordingly, and the maximum sensitivity for this assay of AFP-L3 was set to 0.605ng/ml.
6
Comprehensive Metabolic and Microbiome Assessment
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Physical examinations were performed including body weight, body height, waist and hip circumference. Body mass index (BMI) was calculated as weight divided by the square of height (kg/m2). VAT was measured by a multi-frequency bioelectrical impedance analyzer (Inbody 770, Biospace Co. Ltd.). Venous blood samples were collected in the morning following an overnight fast. An auto-biochemical analyzer (Roche/Hitachi Cobas C501, Roche Diagnostic Corp, Indianapolis) was employed to determine plasma concentrations of triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL). Fasting blood glucose (FBG) and fasting insulin (FINS) were examined using electrochemiluminescent immunoassay on the E601 module of the Roche Cobas 6000 (Burgess Hill, West Sussex, U.K.). HOMA-IR was calculated using the following equation: HOMA-IR = insulin × glucose/22.5. Glycosylated hemoglobin (HbA1c) concentrations were quantified using high-performance liquid chromatography (HPLC) with a D10 set (Bio-RAD, Hercules, California). Fecal samples were collected and stored frozen at −80°C until further analysis and then sent to Beijing Genomics Institute to extract DNA and identify the zygosity.
7
Serum AMH and E2 Measurement
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One sample of 2 ml of blood was collected from each rat. The blood samples were centrifuged, and the serum was separated into two parts: one part was used for AMH analysis, and one part was used for E2 analysis. The serum levels of AMH and E2 at the indicated time points (1, 2, 3, and 5 weeks after single- or multiple-injection hPD-MSC therapy) were measured using the Elecsys® AMH immunoassay (Roche Diagnostics GmbH, Mannheim, Germany) and Elecsys® Estradiol III (Roche Diagnostics GmbH), respectively. All serum markers (AMH and E2) were determined in a single measurement on the e601 module of the fully automated Cobas 6000 system (Roche Diagnostics GmbH). Assays were performed according to the manufacturer’s instructions.
8
Quantification of AFP-L3 Biomarker
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Blood samples were collected when diagnosed and before any interventions, then were frozen at -80°C before use. The levels of AFP and AFP-L3 were detected using AFP testing reagents supplied by Roche (Germany) through an electrochemical luminescence analyzer (e601 module, Roche, Germany). Prior to AFP-L3 detection, the blood samples need to be pre-processed. Every AFP-L3 portion supernatant was split from the blood samples via a Hotgen Biotech glycosyl capture spin-column. The primary mechanism of the spin-column was that the spin-column first pre-loaded Lens culinaris agglutinin to have the fucosylated stuff thoroughly captured, followed by a simple sugar added to compete with the aforementioned captured fucosylated stuff to bind to Lens culinaris agglutinin. Hence, the fucosylated stuff in the sera, the AFP-L3 included, was consequently eluted from the spin-column. After cleaning, free AFP was removed, and AFP-L3 was obtained by elution and centrifugation. Then the Roche electrochemical luminescence analyzer was applied to detecting the contents of total AFP and AFP-L3 respectively. the AFP-L3 percentages were calculated accordingly, and the maximum sensitivity for this assay of AFP-L3 was set to 0.605ng/ml.
9
Hormonal Responses to Exercise Training
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Blood samples from each participant were collected at phase A, B and C, at 07:00-09:00 h, 48-72 h following the last exercise session as previously described (23, (link)24, (link)25, 26) (link). Serum concentrations of TT, sex hormonebinding globulin (SHBG) and cortisol were determined by electrochemiluminescent immunoassay on the E601 module of the Roche Cobas 6000. Inter-assay CVs over a six-month period were 4.5, 2.4 and 4.2% for TT, SHBG and cortisol respectively. Analyses were carried out in a clinical pathology laboratory (Royal Glamorgan Hospital, Wales, UK). Free-T was calculated using the Vermeulen formula (27) (link), which has been validated against equilibrium dialysis (28) (link). The testosterone:cortisol ratio (T:C) was calculated by the following equation: T:C = 100•(TT ÷ cortisol).
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