Blood collected in EDTA-coated tubes were immediately stored on ice, centrifuged and plasma was stored at −80 °C until analyses. Blood collected in serum-tubes was stored at room temperature for at least 30 min to allow coagulation, followed by centrifugation and storage at −80 °C until analyses. Glucose (Hk-CP, Axonlab, Amsterdam, The Netherlands) and FFA (NEFA-HR, WAKO chemicals, Neuss, Germany) in meal test samples were analyzed enzymatically in EDTA plasma using a Pentra 400 (Horiba, Montpellier, France). Insulin during the meal test was analyzed using RIA, and insulin during the hyperinsulinemic euglycemic clamp was analyzed using ELISA. Triglycerides (Sigma, Zwijndrecht, The Netherlands), cholesterol (CHOD-PAP, Roche Diagnostics, Mannheim, Germany), and HDL-cholesterol (CHOD-PAP, Roche Diagnostics, Mannheim,Germany) after precipitation of apoB-containing lipoproteins with phosphotungstic acid and magnesium ions, were analyzed in serum also using a Pentra 400. LDL-cholesterol was calculated according the Friedewald equation38 (link).
Chod pap
Manufactured by Roche
Sourced in Germany, Switzerland, Netherlands, United States
The CHOD-PAP is a laboratory equipment product used for the enzymatic colorimetric determination of cholesterol levels in biological samples. It provides a reliable and efficient way to measure cholesterol concentrations.
Automatically generated - may contain errors
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72 protocols using chod pap
1
Plasma and Serum Analyses Protocol
2
Comprehensive Metabolic and Inflammatory Panel
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Glucose (Hk-CP, Axonlab) and free fatty acids (FFAs) (NEFA-HR, WAKO Chemicals) were analyzed enzymatically in EDTA plasma using a Pentra 400 (Horiba). Triglycerides (Sigma), cholesterol (CHOD-PAP, Roche Diagnostics), and HDL cholesterol (CHOD-PAP, Roche Diagnostics), after precipitation of apoB-containing lipoproteins with phosphotungstic acid and magnesium ions, were analyzed in serum also using a Pentra 400. All samples from 1 subject were analyzed within 1 run. LDL cholesterol was calculated for 12 participants according to the Friedewald equation (40 (link)). In a subset of 7 participants (mean ± SD age: 60 ± 3 y; BMI: 30.0 ± 1.7; n = 2 women) inflammatory cytokine concentrations were measured on a Luminex® 200™ system using an inflammation 20-plex human Procartaplex panel (eBioscience, EPX200-12185-901) containing markers for sE-Selectin; intercellular adhesion molecule 1/CD54; IL-1α; IL-4; IL-12p70; IL-17A/cytotoxic T lymphocyte antigen -8; IP-10/CXC chemokine ligand 10; monocyte chemoattractant protein-1/CC chemokine ligand (CCL) 2; macrophage inflammatory protein (MIP) -1α/CCL3; MIP-1β/CCL4; sP-Selectin; and TNF-α.
3
Cardiovascular Risk Factors Assessment
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Baseline information was collected by physical examination, nonfasting blood samples and self‐administered questionnaires. Blood pressure was recorded with an automatic device (Dinamap Vital Signs Monitor 1846; Critikon Inc) by trained personnel. Participants rested for 2 minutes in a sitting position and then 3 readings were taken on the upper right arm at 2‐minute intervals. The average of the 2 last readings was used in the analysis. Nonfasting blood samples were collected from an antecubital vein. Serum was prepared by centrifugation after a 1‐hour respite at room temperature and analyzed at the Department of Clinical Biochemistry, University Hospital of North Norway. Serum total cholesterol was analyzed by an enzymatic colorimetric method using a commercially available kit (CHOD‐PAP, Boehringer‐Mannheim). Serum HDL‐cholesterol was measured after precipitation of lower‐density lipoproteins with heparin and manganese chloride. Body mass index (BMI) was calculated as weight in kilograms divided by the square of height in meters (kg/m2). History of arterial cardiovascular disease (CVD) (angina pectoris, myocardial infarction, or stroke), diabetes and current smoking was obtained from a self‐administered questionnaire.
4
Quantification of Serum Biomarkers in Nephrotic Syndrome
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Serum creatinine was measured by the isotope dilution mass spectrometry–traceable compensated method (Hitachi Modular P-800; Roche Diagnostics, Mannheim, Germany). The estimated glomerular filtration rate was calculated using the Chronic Kidney Disease Epidemiology Collaboration equation.12 (link) For lipid measurement, blood was collected from the antecubital vein in glass tubes with no additives after an overnight fast of 12 hours. High-density lipoprotein cholesterol and LDL-C were isolated by sequential ultracentrifugation. In the 2 lipoprotein subfractions, cholesterol was determined using enzymatic methods (CHOD-PAP; Boehringer Mannheim, Mannheim, Germany). Albumin concentration was determined using the green bromocresol method. Plasma PCSK9 concentrations were quantified using plasma samples obtained at the time of diagnosis of nephrotic syndrome and 6 months after starting treatment with PCSK9 inhibitors, using sandwich enzyme-linked immunosorbent assays according to the manufacturer’s instructions (R&D Systems, Minneapolis, MN, USA).
5
Metabolic Risk Factors and CVD
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Total cholesterol and high-density lipoprotein cholesterol were measured as mg/dL in serum by enzymatic methods (CHOD-PAP, Boehringer Mannheim, Germany) and dyslipidemia was defined as ratio of total cholesterol to high-density lipoprotein cholesterol ≥ 5.0. Type 2 diabetes was self reported and confirmed by a physician diagnosis. Trained medical staff measured the body weight (kg) and height (cm) of all participants anthropometrically as part of a standardized medical examination. Calibration of measuring instruments was ensured through weekly or daily inspections using standard weights. Body mass index (BMI) was calculated as weight in kilograms divided by height in meters squared and obesity was defined as having a BMI ≥ 30 kg/m². History of parental hypertension was based on participant’s report of either parent with hypertension. Cardiovascular (CVD) conditions were based on a self-reported history of myocardial infarction, heart failure, angina, or stroke.
6
Serum Biomarkers Measurement Protocol
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Serum total Cho and TG were routinely measured in the Laboratory Department of the General Hospital of people’s Liberation Army by enzymatic methods with a commercial kit (CHOD-PAP, Boehringer-Mannheim GmbH, and Konelab TRIGLYCERIDES, Thermo Electron Co.). Serum concentrations of IFN-γ, TNF-α and IL-6 were measured by using enzyme-linked immunosorbent assay (ELISA) kits from R&D systems according to manufacturer’s instructions.
7
Standardized Cardiometabolic Assessments
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All participants underwent a standardized examination and interview, as detailed elsewhere [11 ]. Briefly, systolic blood pressure was measured according to the WHO MONICA manual. Total and HDL-cholesterol were determined from non-fasting blood samples by enzymatic methods (CHOD-PAP, Boehringer Mannheim, Ingelheim am Rhein, Germany) according to the MONICA manual. Weight and height were measured by standardized scales and BMI was calculated as weight in kg divided by height in m2. Obesity was defined as a BMI ≥ 30 kg/m2. Smoking and physician-diagnosed diabetes were assessed by self-report.
8
Venous Blood Sampling for Cholesterol Analysis
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Non-fasting venous blood samples were collected by standard methods by trained personnel, with the participant sitting. A brief venous stasis applied to the upper arm was released before venipuncture. Serum total cholesterol concentrations were analysed within 48 hours by CHOD-PAP enzymatic colorimetric methods with commercial kits (Boehringer-Mannheim (Tromsø 2–4) and Roche Diagnostics (Tromsø 5–7), Mannheim, Germany) at the Department of Laboratory Medicine, University Hospital of North Norway, Tromsø. Use of LLD was assessed through questionnaires (Tromsø 4–7, except participants ≥70 years in Tromsø 4) and a written list of brand names of medications used on regular basis checked by health personnel at the study site (in Tromsø 4, only in individuals aged 55–74 years and 5.5% of participants >74 years). Thus, 1362 participants 75–97 years were not asked about LLD use in Tromsø 4. Self-report of medication used regularly for chronic conditions is considered accurate.11 (link) A comparison of data from Tromsø 6 and information from the Norwegian Prescription Database (time window 180 days) showed a kappa value of 0.94 (95% CI 0.93 to 0.95) for LLD (A E Eggen, unpublished data, 2017).
9
Cardiovascular Risk Factors Assessment
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Blood pressure was measured on the right arm in a sitting position using a Hawksley random-zero sphygmomanometer, and three were taken half an hour after the clinical interview in 3-min intervals. Blood pressure was assessed by obtaining the average of the latter two repeated-blood pressure measurements, and hypertension was defined as ≥ 140/90 mmHg and/or use of antihypertensive medication31 (link). Total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C) were measured as mg/dL in serum by enzymatic methods (CHOD-PAP, Boehringer Mannheim, Germany) and dyslipidemia was defined as the ratio of total cholesterol to high-density lipoprotein cholesterol ≥ 5.032 (link). Body mass index (BMI) was calculated as weight in kilograms divided by height in meters squared and obesity was defined as having a BMI ≥ 30 kg/m233 (link).
10
Nephrectomy-induced Uremia in Mice
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