At the screening examination, portions of the plasma specimens were stored at −80°C until serum sdLDL cholesterol concentrations were measured in 2014 on a Hitachi 7180 automated chemistry analyzer using a homogeneous assay (sdLDL-EX "SEIKEN"; Denka Seiken, Tokyo)15 ). The subjects were divided into two groups, those with serum sdLDL cholesterol of < 35 and of ≥ 35 mg/dL, which is the cutoff value reported previously12 ).
Sdldl ex
Manufactured by Denka Seiken
Sourced in Japan
The SdLDL-EX is a laboratory equipment used for the analysis of small dense low-density lipoprotein (sdLDL) particles. It provides accurate measurements of sdLDL levels in biological samples.
Automatically generated - may contain errors
8 protocols using sdldl ex
1
Serum small dense LDL Measurement
2
Fasting Biomarker Measurement for Cardiovascular Risk
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Fasting serum samples for sdLDL-C measurement were collected at baseline and 6 months after randomization. They were stored at −70℃ or colder. Serum sdLDL-C level was directly measured on a LABOSPECT 006 automated chemistry analyzer using a homogenous assay (sdLDL-EX “SEIKEN;” Denka Seiken, Tokyo, Japan)26)
. The central laboratory measured serum triglycerides, total cholesterol, and high-density lipoprotein cholesterol (HDL-C) levels. Serum LDL-C was calculated with the Friedewald formula27)
unless the triglyceride level was ≥ 400 mg/dL, when LDL-C was directly measured by a homogenous assay. Non-HDL-C was defined as total cholesterol minus HDL-C. TRL-C level was defined as the difference between non-HDL-C and LDL-C. These calculated TRL-C levels have been reported to closely approximate triglyceride levels28
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Fasting serum samples for sdLDL-C measurement were collected at baseline and 6 months after randomization. They were stored at −70℃ or colder. Serum sdLDL-C level was directly measured on a LABOSPECT 006 automated chemistry analyzer using a homogenous assay (sdLDL-EX “SEIKEN;” Denka Seiken, Tokyo, Japan)26)
. The central laboratory measured serum triglycerides, total cholesterol, and high-density lipoprotein cholesterol (HDL-C) levels. Serum LDL-C was calculated with the Friedewald formula27)
unless the triglyceride level was ≥ 400 mg/dL, when LDL-C was directly measured by a homogenous assay. Non-HDL-C was defined as total cholesterol minus HDL-C. TRL-C level was defined as the difference between non-HDL-C and LDL-C. These calculated TRL-C levels have been reported to closely approximate triglyceride levels28
,
29)
.
3
Quantifying Serum Small Dense LDL
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At the screening examination, the portions of the plasma specimens were stored at −80°C until serum sdLDL cholesterol concentrations were measured in 2014. Serum sdLDL cholesterol concentrations were directly measured on a Hitachi 7180 automated chemistry analyzer using a homogeneous assay (sdLDL-EX “SEIKEN”; Denka Seiken, Tokyo)17 (link), 18 (link)), which received Food and Drug Administration clearance on August 18, 2017. The subjects were divided into four groups according to the quartiles of serum sdLDL cholesterol levels: Q1, ≤ 24.4 mg/dL [≤ 0.63 mmol/L]; Q2, 24.5–32.8 mg/dL [0.63–0.84 mmol/L]; Q3, 32.9–43.6 mg/dL [0.85–1.12 mmol/L]; and Q4, ≥ 43.7 mg/dL [≥ 1.13 mmol/L].
4
EMPATHY Study Lipid Profile Analysis
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In the EMPATHY study, blood samples were collected at each clinic from all patients at registration, and every year after registration, for analysis of lipid profile: TC, LDL-C, HDL-C, TG, ApoA1, ApoB, and sdLDL-C. TC levels were measured using the chemical oxygen demand method. LDL-C, HDL-C, and sdLDL-C levels were measured with direct homogenous assays using detergents (LDL-EX, HDL-EX, sdLDL-EX, Denka Seiken, Tokyo, Japan). TG levels were examined by the enzymatic assay of glycerol-3-phosphate oxidase. Apolipoprotein levels were measured using the turbidimetric immunoassay. All assays were performed at SRL (Tokyo, Japan). TRL-C was calculated as TC minus HDL-C minus LDL-C
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In the EMPATHY study, blood samples were collected at each clinic from all patients at registration, and every year after registration, for analysis of lipid profile: TC, LDL-C, HDL-C, TG, ApoA1, ApoB, and sdLDL-C. TC levels were measured using the chemical oxygen demand method. LDL-C, HDL-C, and sdLDL-C levels were measured with direct homogenous assays using detergents (LDL-EX, HDL-EX, sdLDL-EX, Denka Seiken, Tokyo, Japan). TG levels were examined by the enzymatic assay of glycerol-3-phosphate oxidase. Apolipoprotein levels were measured using the turbidimetric immunoassay. All assays were performed at SRL (Tokyo, Japan). TRL-C was calculated as TC minus HDL-C minus LDL-C
5
Detailed Metabolic Profile in Patients
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The demographic data including age, sex, medical history, body mass index (BMI), and laboratory data before the treatment were collected. BMI was calculated by dividing patients' weight (kg) by the square of their height (m). Laboratory data included alanine 2-oxoglutarate aminotransferase (ALT), serum albumin, LDL, high-density lipoprotein cholesterol (HDL), TG, and alpha-fetoprotein (AFP). The fibrosis-4 (FIB-4) index was calculated using the following equation: [age (years)×aspartate 2-oxoglutarate aminotransferase (U/L)]/[platelet count (104/μL)×10×ALT (U/L)1/2]. Hypertension and diabetes mellitus were determined by the medical history, using of medications, and the diagnostic criteria. Hypertension was defined as blood pressure ≥140/90 mmHg (23 (link)). Diabetes mellitus was defined as a fasting blood glucose ≥126 mg/dL or hemoglobin A1c ≥6.5% (23 (link)). SdLDL was measured using a homogeneous assay (sd LDL-EX, Denka Seiken, Tokyo, Japan). Biochemistry tests including those for sdLDL were performed on BioMajesty JCA-BM6070 (JEOL, Tokyo, Japan).
6
Detailed Lipid Biomarker Measurement Protocol
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A central committee, composed of the chief medical officers of all 13 participating districts, developed a detailed manual for data collection. Body weight was recorded with the subjects clothed. Height was measured with stockinged feet. Body mass index (BMI) was calculated as weight (kg)/height (m2). Blood samples were taken after overnight fasting. TC was measured via a cholesterol dehydrogenase-ultraviolet method. Triglycerides (TG) was measured using an enzymatic method. LDL-C and high-density lipoprotein cholesterol (HDL-C) were measured by direct methods using a commercial kit (Cholestest from Sekisui Medical, Tokyo, Japan). SdLDL-C level was directly and selectively measured using a commercial kit (sdLDL-EX from Denka Seiken, Tokyo, Japan). An external laboratory (SRL, Tokyo, Japan) measured the serum lipid markers. The markers were measured by the standardised programme proposed by the Clinical and Laboratory Standards Institute. The nonHDL-C was calculated by subtracting HDL-C from TC. Information about medical history, lifestyle and menopausal status were obtained with a self-reported questionnaire. Smoking status was classified as smoking, former smoking, or never smoking.
7
Lipid Profile Measurement Protocol
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In the EMPATHY study, blood samples were collected at each clinic from all patients at registration, and every year after registration, for analysis of lipid pro le: TC, LDL-C, HDL-C, TG, ApoA1, ApoB, and sdLDL-C. TC levels were measured using the chemical oxygen demand method. LDL-C, HDL-C, and sdLDL-C levels were measured with direct homogenous assays using detergents (LDL-EX, HDL-EX, sdLDL-EX, Denka Seiken, Tokyo, Japan). TG levels were examined by the enzymatic assay of glycerol-3-phosphate oxidase.
Apolipoprotein levels were measured using the turbidimetric immunoassay. All assays were performed at SRL (Tokyo, Japan).
Apolipoprotein levels were measured using the turbidimetric immunoassay. All assays were performed at SRL (Tokyo, Japan).
8
Lipid Profile Measurement Methods
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Total cholesterol, TG, HDL-C, and LDL-C levels were assayed by enzymatic techniques. RLP-C was measured in triplicate after isolation by using an immunoaffinity gel containing monoclonal antibodies to human apolipoprotein B-100 and apolipoprotein A-I (Jimro-II RLP-C kit, Otsuka, Japan). Serum MDA-LDL levels were measured by a sandwich enzyme-linked immunosorbent assay (ELISA) system (Sekisui Medical, Tokyo, Japan) and sd-LDL-C was measured using a novel detergent-based homogenous assay in a commercially available kit (sd-LDL-EX ''Seiken,'' Denka Seiken, Tokyo, Japan).
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