Fasting blood samples of all participants were collected from the cubital vein between 7:00 a.m. and 9:00 a.m. Ethylenediaminetetraacetic acid plasma samples for the measurement of serum ACTH were stored at 2°C to 4°C before the test. Serum ACTH and DHEAS levels were measured by chemiluminescence on an Immulite 2000 analyzer (Siemens Healthcare Diagnostics Inc., Munich, Germany). The normal reference values of ACTH and DHEAS at 8:00 a.m. were <10.12 pmol/L and 350.0–4300.0 µg/L, respectively. Serum cortisol, T, LH, and FSH levels were measured by chemiluminescence on an ADVIA Centaur XP Analyzer (Siemens Healthcare Diagnostics, Inc., Co Dublin, Ireland); and normal reference values were as follows: cortisol 198.7–797.5 nmol/L, T 8.4–28.7 nmol/L, LH 1.5–9.3 mU/ml, and FSH 1.4–18.1 U/L. Other laboratory examinations included the tests for alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (GGT), fasting blood glucose (FBG), total cholesterol (TC), total triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and creatinine (Cr) levels. All the processes of drawing blood were successful without any stress. All assays were performed according to the manufacturers' recommendations. All biochemical determinations were conducted in the same laboratory with standard methods.
Immulite 2000 analyzer
Manufactured by Siemens
Sourced in United States, Germany, United Kingdom
The Immulite 2000 analyzer is an automated immunoassay system designed for in vitro diagnostic testing. It provides automated sample handling, incubation, and measurement of various analytes from patient samples.
Automatically generated - may contain errors
Lab products found in correlation
Advia centaur analyzer, by Siemens (7 mentions)
Advia centaur xp analyzer, by Siemens (2 mentions)
Immulite 2000, by Siemens (2 mentions)
Insulin, by R&D Systems (2 mentions)
Leptin, by Thermo Fisher Scientific (2 mentions)
Bs 200 chemistry analyzer, by Mindray (2 mentions)
Nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (2 mentions)
Dimension vista analyzer, by Siemens (1 mentions)
Coated tube ria, by DiaSorin (1 mentions)
Cobas e, by Roche (1 mentions)
Symbia t6, by Siemens (1 mentions)
Biotek synergy lx, by Agilent Technologies (1 mentions)
Agilent bioanalyzer 2100 system, by Agilent Technologies (1 mentions)
Agilent rna 6000 pico kit, by Agilent Technologies (1 mentions)
Cobas 8000 platform, by Roche (1 mentions)
Dimension vista 1500 analyzer, by Siemens (1 mentions)
Bn 2 equipment, by Siemens (1 mentions)
Multiscan fc plate reader, by Thermo Fisher Scientific (1 mentions)
Elisa kit, by USCN (1 mentions)
Dimension rxl equipment, by Siemens (1 mentions)
40 protocols using immulite 2000 analyzer
1
Comprehensive Hormonal and Metabolic Profiling
2
Hormonal Changes in rTMS for Depression
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The HAM-D and Montgomery-Åsberg Depression Rating Scale (MADRS) were used to assess depressive symptoms. A therapeutic response was defined as a HAM-D reduction or MADRS reduction $50% relative to the pretreatment baseline.
The first and second blood samples were collected between 9:00 a.m. and 10:00 a.m. We took these blood samples within 15 minutes before and after the first and the 10th rTMS sessions, respectively. The third blood sample was taken 4 weeks later at 9:00 a.m. (Figure 1). Blood samples were centrifuged, and the serum was frozen at -20°for future analysis. Thyroid-stimulating hormone (TSH), free triiodothyronine (fT3), and free thyroxine (fT4) were quantified on a Dimension Vista analyzer with dedicated reagents (Siemens, Saint Denis, France). Cortisol, prolactin, growth hormone (GH), and estrogen were quantified on an Immulite 2000 analyzer with dedicated reagents (Siemens). Testosterone was quantified using a radioimmunoassay.
The first and second blood samples were collected between 9:00 a.m. and 10:00 a.m. We took these blood samples within 15 minutes before and after the first and the 10th rTMS sessions, respectively. The third blood sample was taken 4 weeks later at 9:00 a.m. (Figure 1). Blood samples were centrifuged, and the serum was frozen at -20°for future analysis. Thyroid-stimulating hormone (TSH), free triiodothyronine (fT3), and free thyroxine (fT4) were quantified on a Dimension Vista analyzer with dedicated reagents (Siemens, Saint Denis, France). Cortisol, prolactin, growth hormone (GH), and estrogen were quantified on an Immulite 2000 analyzer with dedicated reagents (Siemens). Testosterone was quantified using a radioimmunoassay.
3
Glucose, Insulin, and Incretin Measurement
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Glucose was measured bedside using the glucose oxidase technique (YSI model 2300 STAT Plus, Yellow Spring Instruments, Yellow Spring, OH). Serum insulin and C-peptide concentrations were determined using AutoDELFIA fluoroimmunoassay, Wallac OY, Turku, Finland6 (link), or the Immulite 2000 analyzer, Siemens Healthcare Diagnostics, Tarrytown, NY9 (link),17 (link)) analyzed in one batch per cohort. Similarly, incretins were measured in one batch (e.g. including re-analysis of preoperative and 1-year samples) in Cohort 1 only, due to lack of plasma to reanalyze in cohort 2. For the same reasons, incretins were only analyzed in the fasting state and at one timepoint postprandially (T = 45 min for GLP-1 and T = 60 min for GIP). Total GLP-1 and GIP was analyzed as previously described18 (link), using a radioimmunoassay (RIA, antiserum no. 89390) specific for the C-terminal of the GLP-1 molecule and reacting equally with intact GLP-1 and the primary (N-terminally truncated) metabolite; Total GIP was assayed using the C-terminally directed antiserum #867, which equally recognizes both intact GIP (1–42) and the primary metabolite, GIP (3–42)19 (link).
4
Plasma Glucose and Gut Hormone Analysis
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Blood was collected into pre-chilled Ethylenediamine tetraacetic acid (EDTA)-coated tubes for plasma glucose analysis, centrifuged at room temperature for 45 s and analyzed bedside using the glucose oxidase method (YSI 2300 STAT Plus; YSI, Yellow Springs, OH, United States). Samples for serum C-peptide and paracetamol analyses were collected in clot activator tubes and left to coagulate at room temperature before centrifugation at 4°C for 10 min. Further blood samples were collected into pre-chilled EDTA tubes containing a specific dipeptidyl peptidase 4 (valine-pyrrolidide, final concentration 0.01 mM, a gift from Novo Nordisk, Bagsværd, Denmark) for the analysis of GLP-1 and GIP. EDTA tubes were centrifuged immediately at 4°C for 10 min. Serum was stored at −80°C and plasma at −20°C until batch analysis.
Serum C-peptide concentrations were determined by Immulite 2000 analyzer (Siemens Healthcare Diagnostics, Tarrytown, NY, United States). Paracetamol concentrations, from the liquid test day only, were analyzed using reagents from SEKISUI® Diagnostics (Abbott, Denmark) (21 (link)). Total GLP-1 and total GIP were measured by radioimmunoassays as previous described (22 (link)–24 (link)). Total ghrelin was measured using a Millipore ELISA kit (cat. no. EZGRT-89K, Billerica, MA, United States) (25 (link)).
Serum C-peptide concentrations were determined by Immulite 2000 analyzer (Siemens Healthcare Diagnostics, Tarrytown, NY, United States). Paracetamol concentrations, from the liquid test day only, were analyzed using reagents from SEKISUI® Diagnostics (Abbott, Denmark) (21 (link)). Total GLP-1 and total GIP were measured by radioimmunoassays as previous described (22 (link)–24 (link)). Total ghrelin was measured using a Millipore ELISA kit (cat. no. EZGRT-89K, Billerica, MA, United States) (25 (link)).
5
Testosterone and SHBG Measurement in Hypogonadism
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Testosterone was measured using coated tube RIA (DiaSorin S. p. A., Salluggia, Italy and Diagnostic Products Corporation, LA) and SHBG with chemiluminescent immunoassay method (Immulite 2000 Analyzer, Siemens Healthcare, Erlangen, Germany). Low levels of total testosterone were defined as testosterone serum levels of less than 11.0 nmol/L [26 (link)]. Hypogonadism was defined as testosterone deficiency with clinical signs of hypogonadism.
6
Evaluating Latex Allergy in Patients
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Peripheral venous blood from 8 NRL-allergic patients (1–8) was collected. All patients had positive skin prick tests (≥3 mm above saline negative control) to latex extract (Stallergenes, Paris, France) and increased levels of IgE specific to an extract of NRL (Immulite 2000 analyzer; Siemens Healthcare, Germany). All patients were no atopic or sensitized to other allergens unrelated to latex. Three of eight tested patients (NRL-1, NRL-5, NRL-7) were positive for specific IgE against recombinant Hev b 5 protein, determined by ELISA. The demographic and clinical characteristics of the patients are reported in Table 1 .
7
Blood Collection and Biochemical Analysis
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Blood collection was performed in a private laboratory and for the biochemical
analyzes the volunteers respected a 12-hour fast. The collection was performed
in a vacuum tube with separator gel without anticoagulant; after collection, the
blood was centrifuged for 10 minutes at 3000 rpm to separate the serum from the
other blood components, which was used for the analyzes.
For the determination of glycemia, triglycerides, and the HDL-cholesterol
fraction, a colourimetric enzyme kit was used in an Autohumalyzer A517
device17 (link) (HUMAN et
al., 2004). CRP was measured using an enzyme ELISA kit: Immulite 2000 analyzer
(Siemens Healthcare Diagnostics).20
analyzes the volunteers respected a 12-hour fast. The collection was performed
in a vacuum tube with separator gel without anticoagulant; after collection, the
blood was centrifuged for 10 minutes at 3000 rpm to separate the serum from the
other blood components, which was used for the analyzes.
For the determination of glycemia, triglycerides, and the HDL-cholesterol
fraction, a colourimetric enzyme kit was used in an Autohumalyzer A517
device17 (link) (HUMAN et
al., 2004). CRP was measured using an enzyme ELISA kit: Immulite 2000 analyzer
(Siemens Healthcare Diagnostics).20
8
Comprehensive Metabolic Biomarker Evaluation
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All biochemical data were measured in our laboratory using standard methods. Glycemia was measured via a hexokinase enzymatic reaction by Cobas 501/502 (Roche Diagnostics, Indianapolis, IN, USA). Insulinemia was measured using an electrochemiluminescence immunoassay by Elecsys immunoanalizer and Cobas e (Roche Diagnostics, Indianapolis, IN, USA). HbA1c was assessed using turbidimetric inhibition immunoassay by Cobas Integra 400 Tina-quant Hemoglobin A1c Gen.2 (Roche Diagnostics, Indianapolis, IN, USA). Serum GH was assessed with a two-site chemiluminescent immunometric assay on the IMMULITE 2000 analyzer (Siemens Healthcare Diagnostics, United Kingdom, UK) with a sensitivity of 0.01 ng/mL. Serum total IGF-I was assayed using a solid-phase, enzyme-labeled chemiluminescent immunometric assay by IMMULITE 2000 (Siemens Healthcare Diagnostics, United Kingdom, UK) with a sensitivity of 20 ng/mL.
9
Biochemical and Hormonal Analysis in Rats
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For the determination of biochemical and hormonal parameters, the rats were food deprived for 8 h before testing. Serum glucose levels were measured using a Mindray BS-200 Chemistry Analyzer (Shenzhen Mindray Bio-Medical Electronics Co., Shenzhen, China). The serum concentrations of insulin and leptin were determined using commercial ELISA kits (insulin: R&D Systems, Minneapolis, MN, USA; leptin: Thermo Fisher Scientific, USA; leptin: Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s instructions. Serum 17-β estradiol level was measured with a chemiluminescent immunometric assay on an IMMULITE 2000 analyzer (Siemens Healthcare Diagnostics, Eschborn, Germany). For the insulin assay, the intra-assay coefficient of variation (%CV) was <10%, and the sensitivity was 5 μlU/mL. For leptin, the %CV was 4.3%, and the sensitivity of the assay was 22 pg/mL. For the 17-β estradiol assay, the %CV was respectively 4.5% and the sensitivity was 2 pg/mL.
10
Thyroid Function Assessments Protocol
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FT3, FT4 and TSH assays were conducted on a fully automated ADVIA Centaur analyzer (Siemens Healthcare Diagnostics, USA). Tg and TgAb were assessed on a fully automated IMMULITE 2000 analyzer (Siemens Healthcare Diagnostics, USA). These assays were based on chemiluminescent reaction principle.
The calibration references for the above indices were: FT3, 3.50–6.50 pmol/L; FT4, 11.50–23.50 pmol/L; TSH, 0.30–5.00 μIU/mL; Tg, 0–55.00 ng/mL; TgAb, 0-40.00 IU/mL. In the current study, TgAb > 40 IU/mL was defined as positive, otherwise negative.
The calibration references for the above indices were: FT3, 3.50–6.50 pmol/L; FT4, 11.50–23.50 pmol/L; TSH, 0.30–5.00 μIU/mL; Tg, 0–55.00 ng/mL; TgAb, 0-40.00 IU/mL. In the current study, TgAb > 40 IU/mL was defined as positive, otherwise negative.
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