An enzymatic method is described for determination of total serum cholesterol by use of a single aqueous reagent. The method requires no prior treatment of sample and the calibration curve is linear to 600 mg/dl. Cholesterol esters are hydrolyzed to free cholesterol by cholesterol ester hydrolase (EC 3.1.1.13). The free cholesterol produced is oxidized by cholesterol oxidase to cholest-4-en-3-one with the simultaneous production of hydrogen peroxide, which oxidatively couples with 4-aminoantipyrine and phenol in the presence of peroxidase to yield a chromogen with maximum absorption at 500 nm. The method is reproducible, and the results correlate well with those obtained by automated Liebermann—Burchard procedures (AA-2 and SMA 12/60) and the method of Abell et al. The present method affords better specificity than those previously reported and has excellent precision.

I describe the characterization, extraction, and purification of a cholesterol:oxygen oxidoreductase (EC 1.1.3.6) from Nocardia sp. This enzyme catalyzes oxidation of cholesterol to Δ4-choIestenone, with production of hydrogen peroxide. It is very stable, active over a wide pH range, and has a Km of 1.4 x 10-5 mol/ liter. It is highly specific for Δ4- or Δ5-3β-hydroxycholestanes, and may be applied to the assay of serum total cholesterol. In the procedure presented here, hydrogen peroxide is measured by reaction with quadrivalent titanium and xylenol orange. This constitutes a one-enzyme assay with stable reagents, which does not require protein precipitation and is not subject to interference from hemoglobin or bilirubin.

Lower serum uromodulin (umod) levels correlate with decreasing kidney function, but their relationship with coronary artery disease (CAD) is unknown. We examined the relationships between serum umod, and the development of albuminuria, chronic kidney disease (CKD) and progression of coronary artery calcium (CAC) in adults with type 1 diabetes (T1D), over 12-years. Participants with T1D (n=539, mean age of 40±9, HbA1c 7.7±1.2%, 53% women) in the Coronary Artery Calcification in Type 1 Diabetes study were assessed for CAC progression (128-slice spiral CT), urinary albumin to creatinine ratio (UACR) and estimated GFR (eGFR) calculated by CKD-EPI cre-atinine. CKD was defined as eGFR <60mL/min/1.73m 2 , albuminuria as UACR ≥30 mg/g, and CAC progression as a change in the square root transformed CAC volume ≥2.5. Serum umod was measured using immunoassay kits from Meso Scale Discovery. Logistic regression was used to examine the relationship between umod and incident albuminuria, CKD, or CAC progression, controlling for age, sex, diabetes duration, HbA1c, SBP and LDL-C. Integrated discrimination index (IDI) and net-reclassification improvement (NRI) were used to assess the added prediction performance of umod to a model with age, sex, diabetes duration and ABC risk factors. CAC progression was observed in 49%, incident albuminuria in 6% and incident CKD in 6% of the subjects. Serum umod at baseline was associated with CAC progression (OR: 0.66 95% CI [0.47-0.94] per one SD=138.7 mcg/mg), incident albumin-uria (0.27 [0.11-0.65]) and incident CKD (0.39 [0.20-0.76]) in adjusted models. Results were similar for CAC progression when adjusting for baseline eGFR and UACR, and when excluding participants with CKD or albuminuria. The addition of umod to the baseline model improved relative IDI for CAC progression by 6% (p=0.008) and correctly reclassified 22% cases of CAC progression (p=0.02). In conclusion, lower serum umod levels independently predict development of CAC in adults with T1D. Several genetic variants associated with CVD have been identified in the general population. Our goal was to investigate whether these variants predicted the incidence of major adverse cardiovascular events (MACE) among type 2 diabetes (T2D) subjects at high CVD risk from the ACCORD Study. We analyzed data from 5,360 self-reported white T2D subjects with a median follow-up of 4.7 years. A genetic risk score (GRS) was derived by adding the effect size-weighted number of risk alleles at 40 CVD loci identified in the general population. The association of the GRS with MACE was evaluated by logistic regression including age, gender, and study treatments as covariates. Clinical CVD risk classes were defined as "very high" (presence of CVD at baseline or AHA-ACC 10-year MACE risk score > 15%) or "mod-erately high" (10 y risk <= 15%). The GRS ranged from 27 to 62, with each unit increase being associated with a 3% additional risk of MACE (OR 1.03; 95% CI 1.01-1.06, p=0.002). Subjects in the highest GRS tertile had a 55% increase in MACE risk as co...

Improving dyslipidemia in type 2 diabetic subjects had no effect on the progress of ultrasonically measured arterial disease, although the lower rate of "definite CHD events" in the treated group suggests that this might result in a reduction in the incidence of coronary heart disease.

Polycystic ovary syndrome is associated with biochemical risk factors for premature vascular disease, which cannot be explained by obesity alone.