Background and Aims The prevalence of metabolic dysfunction associated steatotic liver disease (MASLD), formerly known as nonalcoholic fatty liver disease (NAFLD), has become a significant public health concern with an increased atherosclerotic cardiovascular disease risk. This study investigates the impact of NAFLD-related single nucleotide polymorphisms (SNPs) on carotid atherosclerosis development in a Japanese population without diabetes, dyslipidemia, and hypertension. Methods The prospective observational study, part of the Kyushu and Okinawa Population Study (KOPS), included 945 participants (median age 55 [47, 63]) without carotid atherosclerosis, increased alcohol intake, diabetes, dyslipidemia, hypertension, or chronic hepatitis at baseline. NAFLD-related SNPs (GCKR, NCAN, and PNPLA3) were genotyped, and carotid intima-media thickness (cIMT) was measured using ultrasonography. Univariate and multivariate regression analyses were performed to assess the association of NAFLD-related SNPs on newly developed carotid atherosclerosis over five years. Results After five years, 125 (13.2%) participants developed carotid atherosclerosis. The NCAN (rs2228603) T allele was associated with a lower incidence rate of carotid atherosclerosis (4.7% in NCAN CT/TT genotype vs. 13.9% in CC genotype; p = 0.04), and NCAN T allele carriers exhibited a favorable lipid profile. These associations were not altered by either recruiting area or obese. The GCKR T allele and PNPLA3 C allele were associated with low carotid atherosclerosis development rates but were not significant. Conclusions Our results suggested that some NAFLD-related SNPs may influence atherosclerosis through lipid metabolism among Japanese individuals without metabolic syndrome.

BACKGROUND:Low body mass index (BMI) has been associated with marked low-density-lipoprotein-cholesterol (LDL-C) elevations in response to very-low-carbohydrate diets (VLCD). METHODS:We report a case (51-year-old woman, BMI 18.5 kg/m2) whose LDL-C was >500 mg/dL on a VLCD. We characterized her plasma lipoproteins and noncholesterol-sterols (gas chromatography/mass spectrometry) and the DNA sequences of her genes affecting lipid metabolism. We also carried out a large population analysis (224,126 subjects, 54% female, mean age 54 years) examining interrelationships between BMI and serum lathosterol/total cholesterol and β-sitosterol/total cholesterol ratios. RESULTS:In this case, her LDL-C concentration increased from 142 mg/dL to 555 mg/dL on a VLCD, and her plasma β-sitosterol level was very high at 12.8 mg/L. DNA analysis revealed a heterozygous pathogenic ABCG5 exon 9 variant (c.1323_1324+2del at position g.44051049 TACAC>T). With dietary cholesterol restriction and ezetimibe therapy, her LDL-C and β-sitosterol levels decreased by 75% and 46% to 139 mg/dL and 7.1 mg/L, respectively. In the population analysis, we noted a significant inverse correlation between BMI and the plasma β-sitosterol/total cholesterol ratio (r = -0.573, P < .00001). Those with a BMI <20 kg/m2 had mean β-sitosterol/total cholesterol values that were significantly higher (+63%, P < .00001) than values in obese women. The converse was true for the plasma lathosterol/total cholesterol ratio. Similar findings were noted in men. CONCLUSIONS:Our data are consistent with the concepts that low BMI predisposes to increased plasma β-sitosterol/total cholesterol ratios and an increased serum LDL-C when on high cholesterol VLCD, and that this response may be markedly enhanced in subjects with pathogenic heterozygous ABCG5 variants.

Background This study investigated Lp(a) (lipoprotein(a)) levels with heart failure (HF) incidence overall and ejection fraction (EF) subtypes among Black and White participants in a pooled analysis of MESA (Multi‐Ethnic Study of Atherosclerosis), FOS (Framingham Offspring Study), and ARIC (Atherosclerosis Risk in Communities Study). Methods This study was conducted among 16 771 White and Black participants in ARIC (N=10 347), MESA (N=4150), and FOS (N=2274). Baseline was time of Lp(a) measurement (ARIC Visit 4: 1996–1998; MESA Visit 1: 2000–2002; FOS Cycle 6: 1995–1998). HF with reduced EF (HFrEF) was defined as EF <50% and ≥50% as HF with preserved EF (HFpEF). Cox proportional hazards regression was used to evaluate associations between Lp(a) (log‐transformed continuous, dichotomized at ≥30 mg/dL and ≥50 mg/dL, and quartiles) and HF (overall, HFpEF, HFrEF) in the overall population and stratified by race. Analyses were replicated excluding prior history of myocardial infarction. Results There were 2759 HF cases (HFpEF N=859; HFrEF N=649; EF unknown N=1251) through 2019. Among White participants, Lp(a) ≥50 mg/dL was associated with HF risk overall (hazard ratio [HR], 1.19 [95% CI, 1.07–1.34]) and by EF subtype (HFpEF HR, 1.32 [95% CI, 1.08–1.59]; HFrEF HR, 1.33 [95% CI, 1.05–1.67]). Among Black participants, Lp(a) ≥50 mg/dL was not associated with HF risk overall (HR, 0.93 [95% CI, 0.78–1.11]) or by EF subtype (HFpEF HR, 0.97 [95% CI, 0.69–1.35]; HFrEF HR, 0.89 [95% CI, 0.63–1.26]). Associations were no longer significant after excluding prior myocardial infarction. Conclusions Elevated Lp(a) levels are associated with HF risk among White, but not Black individuals, and associations appears to be mostly mediated by a history of myocardial infarction.

BACKGROUND:The substitution of monounsaturated acids (MUFAs) for saturated fatty acids (SFAs) is recommended for cardiovascular disease prevention but its impact on lipoprotein metabolism in subjects with dyslipidemia associated with insulin resistance (IR) remains largely unknown. OBJECTIVES:This study aimed to evaluate the impact of substituting MUFAs for SFAs on the in vivo kinetics of apolipoprotein (apo)B-containing lipoproteins and on the plasma lipidomic profile in adults with IR-induced dyslipidemia. METHODS:Males and females with dyslipidemia associated with IR (n = 18) were recruited for this crossover double-blind randomized controlled trial. Subjects consumed, in random order, a diet rich in SFAs (SFAs: 13.4%E; MUFAs: 14.4%E) and a diet rich in MUFAs (SFAs: 7.1%E; MUFAs: 20.7%E) in fully controlled feeding conditions for periods of 4 wk each, separated by a 4-wk washout. At the end of each diet, fasting plasma samples were taken together with measurements of the in vivo kinetics of apoB-containing lipoproteins. RESULTS:Substituting MUFAs for SFAs had no impact on triglyceride-rich lipoprotein apoB-48 fractional catabolic rate (FCR) (Δ = -8.9%, P = 0.4) and production rate (Δ = 0.0%, P = 0.9), although it decreased very low-density lipoprotein apoB-100 pool size (PS) (Δ = -22.5%; P = 0.01). This substitution also reduced low-density lipoprotein cholesterol (LDL-C) (Δ = -7.0%; P = 0.01), non-high-density lipoprotein cholesterol (Δ = -2.5%; P = 0.04), and LDL apoB-100 PS (Δ = -6.0%; P = 0.05). These differences were partially attributed to an increase in LDL apoB-100 FCR (Δ = +1.6%; P = 0.05). The MUFA diet showed reduced sphingolipid concentrations and elevated glycerophospholipid levels compared with the SFA diet. CONCLUSIONS:This study demonstrated that substituting dietary MUFAs for SFAs decreases LDL-C levels and LDL PS by increasing LDL apoB-100 FCR and results in an overall improved plasma lipidomic profile in individuals with IR-induced lipidemia. TRIAL REGISTRATION:This trial was registered as clinicaltrials.gov as NCT03872349.

Increased cholesterol-rich, low-density, non-calcified atheromas as assessed by computer coronary tomography angiography (CCTA) analyses have been shown to predict myocardial infarction significantly better than coronary artery calcium score or the presence of obstructive coronary artery disease (CAD) as evaluated with standard coronary angiography. Low serum high-density lipoprotein (HDL) cholesterol values are an independent risk factor for CAD. Very small, lipid-poor preβ-1 HDL particles have been shown to be most effective in promoting cellular cholesterol efflux. HDL infusions have been documented to reduce aortic atherosclerosis in cholesterol-fed animal models. However, human studies using infusions of either the HDL mimetic containing recombinant apolipoprotein (apo) A-I Milano or Cerenis Compound-001 with native recombinant apoA-I have been mainly negative in promoting coronary atherosclerosis progression as assessed by intravascular ultrasound. In contrast, a study using 7 weekly infusions of autologous delipidated HDL in six homozygous familial hypercholesterolemic patients was effective in promoting significant regression of low density non-calcified coronary atheroma regression as assessed by computed coronary angiography. This therapy has received Food and Drug Administration approval. Commonwealth Serum Laboratories (CSL) is currently carrying out a large clinical endpoint trial using an HDL complex (native apoA-I with phospholipid), but no final outcome has been reported. Our purpose is to review animal and human studies using various forms of HDL infusion therapy to promote regression of atherosclerosis. In our view, differences in results may be due to: 1) the HDL preparations used, 2) the subjects studied, and 3) the methods used to assess coronary atherosclerosis.

Background/Synopsis Preeclampsia poses a cardiovascular disease risk. Abnormal lipid metabolism is important in the pathogenesis of preeclampsia. Dyslipidemia is strongly associated with atherosclerotic cardiovascular disease (ASCVD) and has a direct effect on endothelial function. Elevated serum or plasma low density lipoprotein cholesterol (LDL-C), small dense LDL-C (sdLDL-C) and lipoprotein (a) [Lp(a)] levels are known independent risk factors for ASCVD, however, extensive lipid changes in preeclampsia are incompletely characterized. Objective/Purpose This study sought to characterize serum and plasma lipoproteins in preeclampsia and control subjects. Methods Frozen serum and plasma from pregnant patients enrolled in the Child Health Advances from Research with Mothers (CHARM) study. Visit 1, 30 cases, 89 controls, gestational age 89 days (36d-247d) and Visit 2, 22 cases, 64 controls, gestational age 189 days (136d-256d), were shipped to Boston Heart Diagnostics for blinded analysis. Patients with gestational diabetes were excluded. Standard serum lipids, direct LDL-C, sdLDL-C, and apolipoprotein (apo) A-I, A-II, and B, and Lp(a) levels were measured by standard chemical methods, and plasma apolipoproteins and particle numbers were assessed using nuclear magnetic resonance (NMR) methodology. Results were adjusted for gestational age, and smoking, and obtained for preeclampsia compared to normal controls - using general linear models for fixed factor analysis. Log transformations were carried out for non-normally distributed parameters. Results In the serum chemistries, significant (p<.05) elevations were found in preeclampsia cases versus controls for direct LDL-C (+10%), small dense LDL-C (+18%), apoB (+10%), and Lp(a) (+50%). Similar alterations were noted in lipoprotein and apolipoprotein analyses by NMR. Results apply in aggregate to all gestational ages. Conclusions There are significant serum and plasma lipoprotein and apolipoprotein alterations seen in preeclampsia cases versus control subjects. Increases in atherogenic small dense LDL particles and their constituents have also been associated with increased ASCVD risk in the general population.

Background: Cerebrotendinous xanthomatosis (CTX) is a rare autosomal recessive lipid disorder. Affected patients often remain undiagnosed until the age of 20-30 years, when they have already developed significant neurologic disease that may not be reversible. An elevated plasma cholestanol concentration has been accepted as a diagnostic criterion for CTX for decades.Objective: Full biochemical characterization was performed for three genetically and clinically confirmed atypical CTX cases with normal plasma cholestanol levels.Methods: Clinical assessment, and genetic/biochemical testing for patients with CTX was performed by their physician providing routine standard of care.Results: We report three new atypical CTX cases with large extensor tendon xanthomas but normal plasma cholestanol levels. All three cases had marked elevations of bile acid precursors and bile alcohols in plasma and urine that decreased on treatment with chenodeoxycholic acid. We also review eight published cases of atypical CTX with normal/near normal circulating cholestanol levels.Conclusion: The atypical biochemical presentation of these cases represents a diagnostic challenge for a disorder for which cholestanol has been believed to be a sensitive biomarker. These cases demonstrate measurements of plasma cholestanol alone are insufficient to exclude a diagnosis of CTX. The data presented is consistent with the concept that bile acid precursors and bile alcohols are sensitive biomarkers for atypical CTX with normal cholestanol, and that such testing is indicated, along with CYP27A1 gene analyses, in patients presenting with significant tendon and/or tuberous xanthomas and/or neurologic disease in early adulthood despite normal or near normal cholesterol and cholestanol levels.

Introduction: The impact of non-alcoholic fatty liver disease (NAFLD) risk SNP variants on ipid metabolism in the general population has yet to be determined. We aimed to assess this association using a large prospective cohort study. Methods: A total of 1488 Japanese men and women (median age 62 years and median body mass index 24.7 kg/m2) free of cardiovascular disease were studied. Among NAFLD risk SNPs, variants of patatin-like phospholipase domain containing 3 (PNPLA3), transmembrane 6 superfamily member 2 (TM6SF2), glucokinase regulator (GCKR), and neurocan (NCAN) were assessed. Plasma total cholesterol, low-density lipoprotein cholesterol (LDL-C), small dense LDL-C (sdLDL-C), LDL-triglycerides (LDL-TG), high-density lipoprotein cholesterol (HDL-C), HDL3-C, triglycerides (TG), lipoprotein(a), and adiponectin were measured at both baseline and after a five-year follow-up. Univariate and multivariate analyses, Kruskal-Wallis, Wilcoxon’s rank sum, and paired t-tests were performed to examine the associations between NAFLD risk SNPs and lipoproteins. Results: All of the measured lipoproteins increased significantly over the 5 years. The TM6SF2 variant did not affect lipoproteins at baseline. However, after 5 years, the non-major allele group had significantly lower HDL-C and HDL3-C and significantly higher LDL-C, sdLDL-C, and LDL-TG than the major allele group. For the GCKR variant, TG, LDL-C, sdLDL-C, and LDL-TG were significantly higher in the non-major allele group at baseline; LDL-C, sdLDL-C, and LDL-TG were still significantly higher in the non-major allele group at 5 years than the CC group. The PNPLA3 and NCAN variants did not affect lipoproteins in this study. Conclusions: NAFLD risk SNP variants might influence atherogenic lipoprotein metabolism.

Introduction: Nonalcoholic fatty liver disease (NAFLD) genetic risk alleles are associated with lipid metabolism. However, the association between those variants and atherosclerosis has not yet been fully evaluated. Hypothesis: We hypothesized that NAFLD risk alleles are associated with the progression of atherosclerosis. Methods: A total of 1,050 Japanese men and women (median age 55 years and median body mass index 22.9 kg/m2) free of cardiovascular disease, dyslipidemia, hypertension, and diabetes were studied. . Among NAFLD risk SNPs, variants of patatin-like phospholipase domain containing 3 ( PNPLA3 ), transmembrane 6 superfamily member 2 ( TM6SF2 ), glucokinase regulator ( GCKR ), and neurocan ( NCAN ) were assessed. Plasma total cholesterol, low-density lipoprotein cholesterol (LDL-C), small dense LDL-C, LDL-triglycerides, high-density lipoprotein cholesterol (HDL-C), HDL3-C, triglycerides, remnant-like particle cholesterol, lipoprotein(a), and adiponectin were also measured. At both baseline and after a five-year follow-up, carotid intima-media thickness (cIMT) was assessed. Atherosclerosis was defined as cIMT ≥ 1.1mm or the presence of a plaque. Univariate and multivariate analyses and chi-square and Fisher’s exact tests were performed to examine the associations between NAFLD risk SNPs, lipoproteins, and progression of atherosclerosis. Results: Both median and maximum cIMT of total participants did not differ between baseline and after a five-year follow-up. Among participants without atherosclerosis at baseline, the rate of plaque development was 13.8% (121/880) with the major allele (CC) of NCAN being significantly higher than other alleles. Variants of NCAN were associated with the development of plaque ( P = 0.04, Fisher’s exact test). Participants with the CT allele of NCAN had significantly lower RLP cholesterol at baseline than those with the CC allele (7.6 mg/dl vs. 10.5 mg/dl, P = 0.02). Variants of PNPLA3 , TM6SF2 , GCKR , and NCAN were not associated with changes in lipoproteins. Conclusions: A NCAN variant, one of the NAFLD risk SNPs, was associated with the development of plaque independent of lipoprotein alterations among healthy Japanese participants.

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in December 2019 and spread rapidly. The purpose of this study was to compare neutralizing an-tibodies (NAbs) following the original booster vaccine in convalescent and naive vaccinated in-dividuals and in a third comparison group consisting of unvaccinated convalescent plasma donors. Methods: We assessed NAbs before and 2 months after a booster vaccine in 68 adults who had completed the initial vaccine series for SARS-CoV-2. Of these subjects, 58 had no history of prior infection (naive vaccinated group) and 10 had been infected with SARS-COV-2 prior to the completing the first vaccine series (convalescent vaccinated group). A third comparison group included unvaccinated convalescent plasma donors (n = 55) from an earlier study with NAbs assessed approximately 2 months after a positive test for SARS-CoV-2. Results: Prior to the booster, convalescent vaccinated subjects had higher NAbs compared to naive vaccinated subjects (p = 0.02). Two months following the booster, NAbs increased in both vaccinated groups. The naive vaccinated group increased more than the convalescent vaccinated group (p = 0.02). NAbs in the naive vaccinated group were almost four times higher than NAbs in the 55 unvaccinated subjects, while the convalescent vaccinated group had levels 2.5 times higher p < 0.01. Conclusion: NAbs in both vaccinated/boosted groups were significantly higher than in the convalescent unvaccinated group (p < 0.01). Our data indicates that subjects with a single infection with SARS-CoV-2 did not have the same levels of neutralizing antibodies that we observed in subjects who were either in the convalescent vaccinated or the naive vaccinated groups.

BACKGROUND:The association between lipoprotein subclasses and carotid intima-media thickness (cIMT) progression has yet to be fully evaluated. We assessed which lipoprotein subclasses were associated with maximum cIMT levels in the general population. METHODS:In this study, cholesterol and triglyceride content of 20 lipoprotein subclasses were analyzed using gel permeation high-performance liquid chromatography (GP-HPLC) in 864 Japanese women and men (mean age 57 y, free of chronic liver or kidney diseases and off lipid-lowering, hormone replacement, or adrenocorticosteroid medications). Univariate and multivariate regression analyses and univariate and partial correlation analyses were performed to examine the relationships between lipoprotein subclasses and maximum cIMT levels. RESULTS:After adjusting for age, sex, systolic blood pressure, smoking, diabetes, and anti-hypertensive agents, elevated low-density lipoprotein (LDL)-2 and -3 cholesterol (particle diameter 25.5 nm and 23.0 nm, respectively; medium and small LDL) were associated with higher maximum cIMT levels in both women and men (all p for trend < 0.05). These associations were significant even after participants taking anti-diabetic or anti-hypertensive agents were excluded. No significant associations were found between any triglyceride subclasses and maximum cIMT levels. CONCLUSIONS:Smaller LDL particle cholesterol values are the most atherogenic lipoprotein parameter.

Serum lipoproteins play a causative role in cardiovascular disease (CVD). Lipoproteins vary in size and density, as well as apolipoprotein (apo), total cholesterol (TC), free cholesterol (FC), triglyceride (TG), and phospholipid (PL) content. High resolution nuclear magnetic resonance (NMR) can simultaneously assess these characteristics within very low-density lipoproteins (VLDL), intermediate-density lipoproteins (IDL), low-density lipoproteins (LDL), and high-density lipoproteins (HDL). We sought to validate the use of specialized NMR analysis and software for the measurement of serum TC, TG, LDL-C, small dense LDL-C, HDL-C, apoA-I, apoB, and LDL particle number (LDL-P). Fasting serum concentrations of TC, TG, direct LDL-C, HDL-C, apoA-I, and apoB were measured in 1,458 men and women (mean age 58.2 years) by 600 MHz NMR using Bruker methods and software and were compared with concentrations obtained by validated automated standardized Roche assays. LDL-P obtained using the Bruker software was compared with that obtained using Numares software. NMR-obtained concentrations were highly correlated with Roche assay results: TC 0.988, TG 0.992, direct LDL-C 0.972, HDL-C 0.969, apoA-I 0.953, and apoB 0.969 (all P< 0.00001). In some cases, corrections were introduced to match absolute concentrations. Correlations between LDL-P analyses were 0.920 ( P< 0.0001). Bruker software provides for assessment of a total of 114 parameters including particle number and TC, FC, TG, PL, and apoB levels in total VLDL, VLDL1-5 (large-small), IDL, and total LDL, LDL1-6 (large-small), and TC, FC, TG, PL, apoA-I, and apoA-II levels in total HDL and HDL1-4 (large-small). However, in our reporting we have focused on the lipid, lipoprotein, and apolipoprotein parameters that we could validate using standard methods, as well as particle number for VLDL, IDL, total LDL, LDL1-6, and HDL and cholesterol and triglyceride values in these particles. We observed substantial gender differences with regard to HDL particles and their constituents. In conclusion, NMR lipoprotein analysis allows for complete lipoprotein analysis except for lipoprotein(a) and holds great promise for CVD risk assessment and for the detection of lipoprotein disorders.