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There is increasing interest in the development of minimally invasive biomarkers for the diagnosis and prognosis of NAFLD via extracellular vesicles (EV). Plasma EVs were isolated by differential ultracentrifugation and quantified by nanoparticle tracking analysis from pre (n = 28) and post (n = 28) weight loss patients. In the pre weight loss group 22 had NAFLD. Nanoplasmon enhanced scattering (nPES) of gold nanoparticles conjugated to hepatocyte-specific antibodies was employed to identify hepatocyte-specific EVs. Complex lipid panel and targeted sphingolipids were performed. Logistic regression analysis was used to identify predictors of NAFLD. Plasma levels of EVs and hepatocyte-derived EVs are dynamic and decrease following NAFLD resolution due to weight loss surgery. Hepatocyte-derived EVs correlate with steatosis in NAFLD patients and steatosis and inflammation in NASH patients. Plasma levels of small EVs correlate with EV sphingolipids in patients with NASH. Hepatocyte-derived EVs measured by the nPES assay could serve as a point-of-care test for NAFLD.

BACKGROUND AND AIMS Alcoholic hepatitis (AH) is diagnosed by clinical criteria, although several objective scores facilitate risk stratification. Extracellular vesicles (EVs) have emerged as biomarkers for many diseases and are also implicated in the pathogenesis of AH. Therefore, we investigated whether plasma EV concentration and sphingolipid cargo could serve as diagnostic biomarkers for AH and inform prognosis to permit dynamic risk profiling of AH subjects. APPROACH AND RESULTS EVs were isolated and quantified from plasma samples from healthy controls, heavy drinkers, and subjects with end-stage liver disease (ESLD) attributed to cholestatic liver diseases and nonalcoholic steatohepatitis, decompensated alcohol-associated cirrhosis (AC), and AH. Sphingolipids were quantified by tandem mass spectroscopy. The median plasma EV concentration was significantly higher in AH subjects (5.38 × 1011 /mL) compared to healthy controls (4.38 × 1010 /mL; P

Cholesterol, triglylceride (TG), and apolipoprotein (apo) B were determined in plasma and in lipoprotein subtractions (VLDL, intermediate-density lipoproteine [IDL], LDL, and HDL) in nonobese NIDDM subjects, who were classified into well-controlled, fairly controlled, or poorly controlled states with or without macrovascular complications (macroangiopathy [MA]). The same analyses were also performed on subjects who had coronary artery disease (CAD) with stable angina pectoris (SA) or unstable angina pectoris (UA) and acute myocardial infarction, cerebrovascular disease (CVD) with atherothrombotic or lacunar infarction, and arteriosclerosis obliterane (ASO). In nonobese NIDDM subjects, the number of apoB-con-taining lipoproteins (VLDL, IDL, and LDL) increased. This alteration was more prominent in subjects with poorly or fairly controlled disease as well as in subjects with MA, but not in those with well-controlled NIDDM. Cholesterol/apoB in LDL decreased in subjects with poorly and fairly controlled diabetes or with MA and was correlated with low HDL cholesterol. The disorder is characterized by hyperbetalipoproteinemia with elevated LDL cholesterol and small dense LDL. In obese NIDDM subjects, the similar disorder was more pronounced. Glycemic control had less effect and hyperinsulinemia, if present, aggravated the lipid disorder. In those with CAD, the number of IDLs increased and the LDL fraction had the properties of small dense LDL. HDL cholesterol decreased. In those with UA, the LDL number increased without elevation of LDL cholesterol, indicating typical hyperbetalipoproteinemia. In subjects with atherothrombotic brain infarction, an increased number of small-sized LDLs was noted. In those with ASO, the number of VLDL and IDL increased with small LDL. HDL cholesterol decreased in those with CAD, cerebrovascular disease, and ASO. Since similar quantitative and qualitative alterations of apoB-containing lipoprotein have been observed in NIDDM patients as well as in those with macrovascular diseases, diabetic patients are thought to be more susceptible to the initiation and progression of atheromatous lesions in coronary, brain, and peripheral arteries.

In order to clarify lipoprotein abnormality in mild to moderate obesity (BMI > or = 25), plasma was separated by table top ultracentrifugation into VLDL (chylomicron), IDL, LDL and HDL. Chol, TG and ApoB were determined in each fraction by enzymatic and sensitive Latex method. The data were analysed according to glucose intolerance and hyperinsulinemia (HI). In obese subjects, irrespective of glucose intolerance, Chol, TG & ApoB levels were high in plasma, and an increase in VLDL (Chol, TG & ApoB), IDL (Chol & ApoB), LDL (Chol & ApoB), and a decrease in HDL-Chol were observed. These levels were also abnormal in nonDM particularly with HI. In DM, HI did not seem to affect hyperlipidemia. Correlation between Chol, TG and ApoB in three ApoB containing lipoprotein subfractions was noted in obesity. The ratio of Chol/ApoB and TG/ApoB in LDL was significantly lower in obesity implying that LDL particles were smaller in size. Half of nonDM patients had HI, and only 29% of DM patients had HI, and both groups had almost the same lipoprotein abnormality. Hyperlipidemia was severe in nonDMHI(+) compared to nonDMHI(-). Therefore, in hyperlipidemia of obesity, hyperinsulinemia plays a role in nonDM and hyperglycemia in DM. Insulin resistance seems to be an important factor in DM. Although the mechanism may be different, the consequence of hyperlipidemia is similar. Increased numbers of ApoB containing lipoproteins and smaller size of LDL are the characteristic features of hyperlipidemia in mild to moderate obesity. Because these quantitative and qualitative changes appear to be linked to an increased risk for premature arteriosclerosis, intensive therapy should be recommended even in mild to moderate obesity.