Individuals had been fasting (12 hours) before their visit. Blood samples were retrieved and processed at a single center and stored at –80°C until use. Interleukin (IL)-6, IL-10 (Bender Medsystems, Burlingame, CA, USA), resistin, and soluble(s)CD14 (R&D Systems, Minneapolis, MN, USA) levels were analyzed using an enzyme-linked immunosorbent assay. Serum ultrasensitive C-reactive protein (us-CRP) was measured by immononephelometry on an IMMAGE analyzer (Beckman-Coulter, Miami, FL, USA). Plasma D-dimer was measured by enzyme-linked fluorescent assay on a VIDAS analyzer (Biomérieux, Marcy-l’Etoile, France). Inducible protein (IP)–10, fractalkine, Monokine induced by gamma interferon (MIG), monocyte chemoattractant protein (MCP)–1, E-selectin, and tumor necrosis factor (TNF)–α were quantified from plasma using the BD Cytometric Bead Array system (BD, Franklin Lakes, NJ, USA).
Immage analyzer
Manufactured by Beckman Coulter
Sourced in United States, France
The IMMAGE analyzer is a laboratory instrument used for the quantitative measurement of specific proteins in biological samples. It employs immunoassay technology to analyze a variety of analytes, including proteins, hormones, and other biomolecules. The IMMAGE analyzer provides accurate and reliable results to support clinical decision-making in various healthcare settings.
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Lab products found in correlation
Il 10, by Thermo Fisher Scientific (2 mentions)
Vidas analyzer, by bioMérieux (2 mentions)
Cytometric bead array system, by BD (2 mentions)
Resistin, by R&D Systems (2 mentions)
Quantikine, by R&D Systems (2 mentions)
Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions)
Soluble s cd14, by R&D Systems (1 mentions)
Architect ci8200 analyzer, by Abbott (1 mentions)
Cobas integra 400 analyzer, by Roche (1 mentions)
Cobas ampliprep cobastaqman hiv 1 assay, by Roche (1 mentions)
Il 18, by Thermo Fisher Scientific (1 mentions)
Cobas c311, by Roche (1 mentions)
6 protocols using immage analyzer
1
Biomarker Analysis in Fasting Participants
2
Metabolic Markers in HIV Treatment
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Clinical and laboratory data were collected before ART initiation and during treatment. Cryopreserved serum and plasma were used for centralized measurements at Tenon Hospital Biochemistry Department (Paris, France). Plasma glucose (hexokinase) and insulin (chemiluminescence immunoassay) were measured on the Architect® Ci8200 analyzer (Abbott). High-sensitivity (hs) CRP was measured by immunonephelometry on an IMMAGE analyzer (Beckman-Coulter). High-sensitivity (hs) IL-6, sCD14, sCD163, sTNFRI and II were measured with enzyme-linked immunosorbent assays (ELISA) (Quantikine®, R&D Systems).
Impaired fasting glucose metabolism was defined, in the absence of diabetes, by at least one fasting glycemia value between 5.6 and 6.9 mmol/l. Diabetes was recorded if the patient was receiving antidiabetic treatment or if the fasting glucose was ≥ 7 mmol/l. ART regimens and treatments for comorbidities were recorded every 6 months. To better take into account actual ART drugs exposure during the 3-year study period, we distinguished never-exposed patients, patients who stopped ART drugs for more than 6 months before the 3-year endpoint, and patients still taking ART.
Impaired fasting glucose metabolism was defined, in the absence of diabetes, by at least one fasting glycemia value between 5.6 and 6.9 mmol/l. Diabetes was recorded if the patient was receiving antidiabetic treatment or if the fasting glucose was ≥ 7 mmol/l. ART regimens and treatments for comorbidities were recorded every 6 months. To better take into account actual ART drugs exposure during the 3-year study period, we distinguished never-exposed patients, patients who stopped ART drugs for more than 6 months before the 3-year endpoint, and patients still taking ART.
3
Body Composition and Metabolic Biomarkers
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Analyses of these variables have been previously described [10 (link)]. Briefly, body weight measurement was conducted using a calibrated balance (Balance Industrielle Montréal, Québec, Canada) while other body composition variables, including fat mass and lean mass, were measured by Dual-energy X-ray absorptiometry (DXA) scans using the LUNAR Prodigy system (software version 6.10.019; General Electric Lunar Corporation, Madison, WI, USA). Height was measured using a wall stadiometer (Perspective Enterprises, Portage, MI, USA) while BMI was calculated as body weight (kg)/height(m)2. Visceral fat and thigh muscle attenuation were measured by computed tomography scan (General Electric Medical Systems, Milwaukee, WI, USA). Adjusting variables were measured using the COBAS INTEGRA 400 analyzer (Roche Diagnostic, Montréal Canada) for high-density lipoprotein cholesterol (HDL-C), triglycerides and glucose. The Friedwald formula was utilised to calculate cholesterol (LDL-C) concentration. C-reactive protein (CRP), haptoglobin, transferrin and orosomucoid levels were measured by immunonephelometry on an IMMAGE analyzer (Beckman Coulter, Villepinte, France).
4
Measuring HIV-Viremia and Inflammatory Markers
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HIV-VL was measured using an adapted Cobas AmpliPrep/Cobas TaqMan HIV-1 assay (Roche Diagnostics, Meylan, France; quantification limit: 1 copy/ml). [7] (link) Patients were defined as having either detectable (D; 1–20 copies/mL, n = 14) or undetectable (UD; <1 copies/mL, n = 33) levels of HIV-viremia.
Inflammatory markers and chemokines were selected for their specific role in HIV-infection [7] (link) and quantified from serum samples stored at −80°C. Interleukin (IL)-6, IL-18, IL-10, IL-27 (Bender Medsystems, Burlingame, CA, USA), resistin (R&D Systems, Minneapolis, MN, USA), total and high molecular weight (HMW) adiponectin (Bühlman, Düsseldorf, Germany) levels were analyzed using an enzyme-linked immunosorbent assay. Serum high-sensitivity C-reactive protein (hs-CRP) and serum amyloid-A (SSA) were measured by immononephelometry on an IMMAGE analyzer (Beckman-Coulter, Miami, FL, USA). Plasma D-dimer was measured by enzyme linked fluorescent assay on a VIDAS analyzer (Biomérieux, Marcy-l′Etoile, France). Chemokines were quantified from plasma using the BD Cytometric Bead Array system (BD, Franklin Lakes, NJ, USA).
Inflammatory markers and chemokines were selected for their specific role in HIV-infection [7] (link) and quantified from serum samples stored at −80°C. Interleukin (IL)-6, IL-18, IL-10, IL-27 (Bender Medsystems, Burlingame, CA, USA), resistin (R&D Systems, Minneapolis, MN, USA), total and high molecular weight (HMW) adiponectin (Bühlman, Düsseldorf, Germany) levels were analyzed using an enzyme-linked immunosorbent assay. Serum high-sensitivity C-reactive protein (hs-CRP) and serum amyloid-A (SSA) were measured by immononephelometry on an IMMAGE analyzer (Beckman-Coulter, Miami, FL, USA). Plasma D-dimer was measured by enzyme linked fluorescent assay on a VIDAS analyzer (Biomérieux, Marcy-l′Etoile, France). Chemokines were quantified from plasma using the BD Cytometric Bead Array system (BD, Franklin Lakes, NJ, USA).
5
Automated Lipid and Inflammatory Biomarkers
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Cholesterol, triglyceride, LDL-C and HDL-C concentrations were determined enzymatically using an autoanalyzer (Cobas c 311, Roche, Germany). HDL-C concentrations were determined after precipitation of apoB-containing lipoproteins by dextran sulfate with polyethylene glycol. LDL-C concentrations were calculated by the Friedewald equation [23] (link). ApoB and high-sensitivity C-reactive protein (hs-CRP) concentrations were determined by nephelometry (Beckman Immage Analyzer, USA).
6
Biomarkers in Inflammatory Conditions
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C-reactive protein (CRP) and SAA protein levels were determined by immunonephelometry using an IMMAGE ® analyzer (Beckman-Coulter, Villepinte, France).
The IL-6 level was assessed by chemiluminescence enzyme immunoassay using a LUMIPULSE ® analyzer (Fujirebio, Tokyo, Japan). The blood levels of other inflammatory cytokines (IL-1β and TNF-α), adipokines (leptin, adiponectin) (Quantikine, R&D Systems, Oxford, UK) and S100A8/A9 proteins (Bühlmann, Amherst, USA) were measured by enzymelinked immunosorbent assay according to the manufacturers' instructions. For these circulating biomarkers, the control group was formed in part by 17 healthy male volunteers from the French study METASPERME (N°IDRCB: 2011-A01052-39) and 7 healthy women volunteers presented in a previous publication [34] . None of the healthy controls were diabetic or obese.
Tryptophan and kynurenine levels were evaluated by high-performance liquid chromatography with a coulometric detection system (ESA Coultronics, ESA Laboratories, Chelsford, MA, USA). The kynurenine/tryptophan ratio was determined from the kynurenine and tryptophan absolute concentrations and used as a marker for IDO activity. The values for our cohort were compared to 48 age-and sex-matched healthy controls.
The IL-6 level was assessed by chemiluminescence enzyme immunoassay using a LUMIPULSE ® analyzer (Fujirebio, Tokyo, Japan). The blood levels of other inflammatory cytokines (IL-1β and TNF-α), adipokines (leptin, adiponectin) (Quantikine, R&D Systems, Oxford, UK) and S100A8/A9 proteins (Bühlmann, Amherst, USA) were measured by enzymelinked immunosorbent assay according to the manufacturers' instructions. For these circulating biomarkers, the control group was formed in part by 17 healthy male volunteers from the French study METASPERME (N°IDRCB: 2011-A01052-39) and 7 healthy women volunteers presented in a previous publication [34] . None of the healthy controls were diabetic or obese.
Tryptophan and kynurenine levels were evaluated by high-performance liquid chromatography with a coulometric detection system (ESA Coultronics, ESA Laboratories, Chelsford, MA, USA). The kynurenine/tryptophan ratio was determined from the kynurenine and tryptophan absolute concentrations and used as a marker for IDO activity. The values for our cohort were compared to 48 age-and sex-matched healthy controls.
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