Casual blood samples were obtained for NHNS 2010. Serum was separated and centrifuged soon after blood coagulation, and plasma samples were collected in siliconized tubes containing sodium fluoride and shipped to a central laboratory (SRL, Tokyo, Japan) for analysis. Serum total cholesterol was measured by the cholesterol dehydrogenase-ultraviolet (UV) method and standardized by the Centers for Disease Control and Prevention/US Collaborating Center for Reference Method Laboratory Network Research in Blood Lipids27 (link)). Triglycerides were measured using enzyme methods, and high-density lipoprotein (HDL) cholesterol and low-density lipoprotein (LDL) cholesterol were measured using direct methods (Cholestest®LDL Sekisui Medical). Information on the use of medications was collected using a self-administered questionnaire. Hypercholesterolemia was defined as total cholesterol level ≥ 6.21 mmol/L (240 mg/dL)28 (link)) and/or the use of antihypercholesterolemic agents. Information on the use of lipid-lowering medications was obtained using a questionnaire and confirmed by trained staff. Untreated hypercholesterolemia was defined as not receiving lipid-lowering medications.
Cholestest ldl
Manufactured by Sekisui
Sourced in Japan
Cholestest LDL is a laboratory equipment product designed to measure the level of low-density lipoprotein (LDL) cholesterol in a sample. It provides a quantitative analysis of LDL cholesterol levels.
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Lab products found in correlation
Cholestest n hdl, by Sekisui (30 mentions)
Pureauto s cho n, by Sekisui (10 mentions)
Pureauto s tg n, by Sekisui (9 mentions)
Automatic analyzer 7600, by Hitachi (4 mentions)
Cholestest cho, by Sekisui (4 mentions)
Hlc 723g9, by Tosoh (3 mentions)
L type tg m test, by Fujifilm (2 mentions)
Nefa hr, by Fujifilm (2 mentions)
Cinq hba1c, by Arkray (2 mentions)
7180 automatic analyzer, by Hitachi (2 mentions)
Micro bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Ldl cholesterol plus 2nd generation, by Roche (2 mentions)
Ldl cholesterol gen 3, by Roche (2 mentions)
Cobas 8000 c702, by Roche (2 mentions)
Cardiophase hscrp, by Siemens (2 mentions)
Jca bm8060, by JEOL (2 mentions)
Jca bm9130, by JEOL (2 mentions)
L type wako cho h, by Fujifilm (2 mentions)
L type wako tg h, by Fujifilm (2 mentions)
Bp 103i2, by Omron (1 mentions)
15 protocols using cholestest ldl
1
NHNS 2010 Cholesterol Measurement
2
Comprehensive Metabolic Profiling in Diabetes
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We measured plasma glucose (PG), hemoglobin A1c (HbA1c; HLC-723G9, TOSOH Co., Ltd., Tokyo, Japan), serum total cholesterol (T-C; Determiner L TC II, Kyowa Medex Co., Ltd., Tokyo, Japan), triglycerides (TG; Determiner L TG II, Kyowa Medex Co., Ltd.), high-density lipoprotein cholesterol (HDL-C; Cholestest N HDL, Sekisui Medical Co., Ltd., Tokyo, Japan), and LDL-C (Cholestest LDL, Sekisui Medical Co., Ltd.). In addition, we measured aspartate transaminase (AST), alanine aminotransferase (ALT), creatinine (Cr), and the urinary albumin-Cr ratio (UACR; N-A TIA MicroALB, Nittobo Medical Co., Ltd., Tokyo, Japan) as a marker for diabetic nephropathy.
3
Comprehensive Metabolic Profiling of Mice
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Blood samples were collected from mice by cardiocentesis. HbA1c levels were measured in whole blood using CinQ HbA1c (ARKRAY). After centrifugation of the blood at 2000 rpm for 20 min at 4°C, the plasma fraction was collected. Plasma samples were used to determine the levels of blood urea nitrogen (UN-S SEIKEN kit; Denka Co., Ltd.), triglycerides (L-type TG M test; Wako), alanine aminotransferase (L-type ALT J2 test; Wako), aspartate aminotransferase (L-type AST J2 test; Wako), total cholesterol (L-type CHO M test; Wako), high-density lipoprotein cholesterol (Cholestest N HDL; SEKISUI Medical Co., Ltd.), low-density lipoprotein cholesterol (Cholestest LDL; SEKISUI Medical Co., Ltd.), creatinine (L-type CRE M test; Wako), non-esterified fatty acid (NEFA-HR; Wako), total ketone bodies (Auto Wako T-KB; Wako) using the Hitachi 7180 automatic analyzer (Hitachi, Tokyo, Japan). Blood insulin levels were analyzed using a mouse insulin ELISA kit (Morinaga Institute of Biological Science).
4
Lipid and Glucose Biomarker Measurement
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After a minimum of 8 hours of fasting, blood samples were obtained. The serum enzyme levels of TC, TG, LDL-C, and HDL-C in the blood samples were measured using a Hitachi Automatic Analyzer 7600 (Hitachi, Tokyo, Japan) after the samples were delivered to the Central Testing Institute in Seoul, Korea. The LDL-C level was calculated using the Friedewald’s formula in subjects with a TG level ≤ 400 mg/dL and was measured using commercially available kits (Cholestest® LDL; Sekisui Medical, Tokyo, Japan) in subjects with a TG level > 400 mg/dL. The fasting blood glucose level was measured after they fasted for 8 hours or more. In addition to fasting blood glucose, the hemoglobin A1c test was also performed. Fasting blood glucose was measured with Tosoh G8 (Tosoh/Japan) using the high-performance liquid chromatography method and with HLC-723G8 HbA1C kit.
5
Cholesterol Measurement by Homogeneous Assay
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The cholesterol levels were measured by a homogeneous assay, Cholestest LDL (SEKISUI Medical, Tokyo, Japan) for LDL-C, CholestestN HDL (SEKISUI Medical) for HDL-C and Cholestest CHO (SEKISUI Medical) for TC. Non-HDL-C was calculated by subtracting HDL-C from TC.
6
Lipid Profile Determination Protocol
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Protein concentrations were determined by the Micro BCA Protein Assay Kit (Thermo Fisher Scientific, Tokyo, Japan). Concentrations of triglycerides (TG), total cholesterol (TC), LDL-cholesterol (LDL-C), and high-density lipoprotein-cholesterol (HDL-C) were quantified using the following commercially available reagents: Quickauto Neo TGII (Shino-Test, Tokyo, Japan), Determiner L TC II (Hitachi Chemical Diagnostic Systems, Tokyo, Japan), Cholestest LDL, and Cholestest N HDL (Sekisui Medical, Tokyo, Japan), respectively.
7
Serum Lipid Measurement Protocol
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Blood samples were drawn from the antecubital vein of each subject the morning after fasting for at least 8 hours. Serum lipid concentrations were directly measured by an enzymatic method using an automated analyzer (Hitachi Automatic Analyzer 7600, Hitachi, Tokyo, Japan), including measurements of total cholesterol (Pureauto S CHO-N; Sekisui Medical, Tokyo, Japan), HDL-C (Cholestest N HDL; Sekisui Medical), and triglycerides (Pureauto S TG-N; Sekisui Medical). Serum LDL-C concentrations (LDL-CD) were directly measured using an enzymatic homogenous assay with Cholestest-LDL (Sekisui Medical). Non-HDL-C was calculated by subtracting HDL-C from total cholesterol. VLDL-C was calculated using the following subtraction equation: VLDL-C = [non-HDL-C]–[LDL-CD].
LDL-CF was estimated as [non-HDL-C]–[triglycerides / 5] [4 ]. LDL-C180 was calculated as [non-HDL-C]–[triglycerides / AF], where AF is an adjustable factor in the 180-cell table described by Martin et al. [9 (link)]. In addition, two LDL-CN estimates were calculated using strata-specific TG:VLDL-C ratios derived from our data set: LDL-C5 based on 5 strata of triglycerides and LDL-C25 based on 25 strata of triglyceride and non-HDL-C levels.
LDL-CF was estimated as [non-HDL-C]–[triglycerides / 5] [4 ]. LDL-C180 was calculated as [non-HDL-C]–[triglycerides / AF], where AF is an adjustable factor in the 180-cell table described by Martin et al. [9 (link)]. In addition, two LDL-CN estimates were calculated using strata-specific TG:VLDL-C ratios derived from our data set: LDL-C5 based on 5 strata of triglycerides and LDL-C25 based on 25 strata of triglyceride and non-HDL-C levels.
8
Standardized Lipid Measurement Protocols
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Serum concentrations of TC, TG, HDL, and LDL were measured using enzymatic methods with an automatic analyzer (Cobas 8000 c702, Roche, Mannheim, Germany). LDL was measured using LDL-cholesterol plus 2nd generation reagent on samples between July 4, 2017, and February 4, 2018, and LDL-cholesterol Gen.3 (Roche, Mannheim, Germany) between February 5 and November 30, 2018. TC, TG, and HDL were measured using Cholesterol Gen.2, TRIGL, and HDL-Cholesterol plus 3rd generation, respectively. The accuracy of lipid measurements was assured through the Accuracy Based Lipid Survey proficiency testing program by the College of American Pathologists and by the Lipids Standardization Program by the Centers for Disease Control, USA [25 (link)].
For population 3 (validation cohort 2, KNHANES 2017), serum TC, TG, HDL, and LDL were measured using enzymatic methods with an automatic analyzer Hitachi 7600–210 (Hitachi, Tokyo, Japan) using PureautoS CHO-N, Pureauto S TG-N, Cholestest N HDL, and Cholestest LDL reagents (Sekisui Medical, Tokyo, Japan), respectively [24 (link)].
For population 3 (validation cohort 2, KNHANES 2017), serum TC, TG, HDL, and LDL were measured using enzymatic methods with an automatic analyzer Hitachi 7600–210 (Hitachi, Tokyo, Japan) using PureautoS CHO-N, Pureauto S TG-N, Cholestest N HDL, and Cholestest LDL reagents (Sekisui Medical, Tokyo, Japan), respectively [24 (link)].
9
Nephrotic Syndrome Biomarker Evaluation
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All of the following assays are routine laboratory tests used to evaluate the clinical condition of patients with nephrotic syndrome: total protein (TP); total cholesterol (TC); triglyceride (TG) (Aqua-auto Kainos TP-II, T-CHO, TG-II Test Kit; Kainos Laboratories); low-density lipoprotein-cholesterol (LDL-C) (Cholestest LDL; Sekisui Medical, Tokyo) and highdensity lipoprotein-cholesterol (HDL-C) (MetaboLead HDL-C; Kyowa Medex, Tokyo). Previously, we confirmed the precision of these assays. 20, 21 High-sensitivity C-reactive protein (hs-CRP) was measured using latex-enhanced immunonephelometry (CardioPhase hsCRP; Siemens Healthcare Diagnostics, NY, USA).
Instead of measuring 24-h urine protein excretion, spot urine protein concentration and its protein/creatinine (P/C) ratio were used to estimate the severity of proteinuria.
Instead of measuring 24-h urine protein excretion, spot urine protein concentration and its protein/creatinine (P/C) ratio were used to estimate the severity of proteinuria.
10
Measuring Blood Biomarkers for Cardiovascular Risk
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Blood samples were collected in the morning after an overnight fast. Plasma sLOX-1 concentrations were measured using a sandwich enzyme-linked immunosorbent assay (ELISA) with two specific monoclonal antibodies against sLOX-1, with recombinant sLOX-1 as the assay standard. In our laboratory, intra-assay and inter-assay CVs for blood sLOX-1 concentration were 1.6–5.2% and 3.9–7.7%, respectively.(5 (link))Serum concentrations of low- and high-density lipoprotein cholesterol (LDL and HDL, respectively) and triglycerides were measured by using commercial enzymatic kits (Cholestest®LDL; Sekisui Medical, Tokyo, Japan: Determiner L HDL-C or MetaboLead HDL-C; Kyowa Medex, Tokyo, Japan: and Determiner C-TG; Kyowa Medex) and an autoanalyzer (JCA-BM 8060; JEOL, Tokyo, Japan). Plasma glucose concentrations and haemoglobin A1c were determined by using a enzymatic kit (L-type Wako Glu2; Wako Pure Chem. Ind., Ltd., Osaka, Japan), a latex agglutination kit (Determiner L HbA1c, Kyowa Medex), and an autoanalyzer (JCA-BM 9130; JEOL). Serum insulin levels were assessed by an electrochemiluminescence immunoassay by using an analyzer (E170; Roche Diagnostics K.K., Tokyo, Japan) and the designated regents.
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