The measurement of the activity of cholesteryl ester transfer protein (CETP), is of high clinical interest and this study reports the use of a direct LDL isolation (d-LDL) technique to determine in one step the amount of radiolabeled cholesteryls esters ([3H]-CE) transferred from exogenous HDL3 to LDL, avoiding the conveniences of the usually used ultracentrifugation or precipitation of apo-B containing lipoproteins in the CETP methodologies. The d-LDL technique providing a specific immunoprecipitation of VLDL, IDL and HDL allowed to directly determine the [3H]-CE transferred on LDL (d-[3H]-CE-LDL). Two methodologies were assayed for the CETP activity using either exogenous or endogenous lipoproteins, and the results with the d-LDL technique were compared with those obtained using the ultracentrifugation (u-[3H]-CE-LDL) considered as the reference method. The intra- and inter-assays were similar in both techniques for the two CETP activity assays. Strong positive correlations were established between values obtained with d-[3H]-CE-LDL and u-[3H]-CE-LDL isolation procedures for CETP activities with exogenous or endogenous lipoproteins (r = 0.972; p = 0.0001 and r = 0.965; p = 0.0001 respectively). In conclusion, the d-LDL technique represents an easy and accurate procedure to measure directly, in normotriglyceridemic plasmas, the amount of [3H]-CE transferred from HDL to LDL by the CETP.

The binding of 125I-labeled lipoproteins to subcellular structures of rat hepatocytes was studied. The protein component of HDL, LDL, and VLDL was found in large and small membranes, nuclei, mitochondria, lysosomes, microsomes, and cell cytosol. The presence in liver nuclear chromatin of proteins immunochemically related to apoA-1 was demonstrated by solid phase immunoenzymatic analysis, dot immunoanalysis, and immunoelectroblotting. The apoprotein A-1 immunoreactivity was detected in transcriptionally inactive total chromatin, transcriptionally active chromatin, and nuclear matrix. The specific apoA-1 immunoreactivity of transcriptionally active chromatin and nuclear matrix was two times as low as that of total and transcriptionally inactive chromatin. Immunoelectroblotting revealed two proteins possessing an apoA-1 immunoreactivity (M(r) 28 and 14 kDa). The 28 kDa protein was found in transcriptionally active chromatin and nuclear matrix, while the 14 kDa one--only in transcriptionally inactive chromatin. the roles of apoproteins in lipid transport to nuclei and the maintenance of chromatin in a transcriptionally active state are discussed.

BACKGROUND

Obestatin is a ghrelin-associated peptide, derived from preproghrelin. Although many of its effects are unclear, accumulating evidence supports positive actions on both metabolism and cardiovascular function. To date, level of obestatin and its correlations to the lipid subfractions in non-diabetic obese (NDO) patients have not been investigated.

METHODS

Fifty NDO patients (BMI: 41.96 ± 8.6 kg/m2) and thirty-two normal-weight, age- and gender-matched healthy controls (BMI: 24.16 ± 3.3 kg/m2) were enrolled into our study. Obestatin level was measured by ELISA. Low-density lipoprotein (LDL) and high-density lipoprotein (HDL) subfractions, intermediate density lipoprotein (IDL) and very low-density lipoprotein (VLDL) levels and mean LDL size were detected by nongradient polyacrylamide gel electrophoresis (Lipoprint).

RESULTS

Serum level of obestatin was significantly lower in NDO patients compared to controls (3.01 ± 0.5 vs. 3.29 ± 0.6 μg/ml, p < 0.05). We found significant negative correlations between the level of obestatin and BMI (r = - 0.33; p < 0.001), level of serum glucose (r = - 0.27, p < 0.05), HbA1c (r = - 0.38; p < 0.001) and insulin (r = - 0.34; p < 0.05). Significant positive correlation was found between obestatin level and the levels of ApoA1 (r = 0.25; p < 0.05), large HDL subfraction ratio and level (r = 0.23; p < 0.05 and r = 0.24; p < 0.05), IDL (r = 0.25 p < 0.05) and mean LDL size (r = 0.25; p < 0.05). Serum VLDL ratio and level negatively correlated with obestatin (r = - 0.32; p < 0.01 and r = - 0.21; p = 0.05). In multiple regression analysis obestatin was predicted only by VLDL level.

CONCLUSIONS

Based on our data, measurement of obestatin level in obesity may contribute to understand the interplay between gastrointestinal hormone secretion and metabolic alterations in obesity.

The effects of dietary fats and phytosterol on the fatty acid composition and lipoprotein cholesterol in serum were studied in female rats, with the following results. (1) The addition of 1% cholesterol to the 20% butter diet decreased the ratio of polyunsaturated fatty acid (PUFA) to saturated fatty acid (SFA) in serum. This phenomenon was negated when there was an intake of cod liver oil and wheat germ oil. (2) When cholesterol was added to the 20% butter diet, the serum total cholesterol increased 3.7-fold, due to an increase in the lower density lipoprotein (LDL + VLDL). (3) The addition of 5% phytosterol to the 10% butter-cholesterol diet reduced the total cholesterol level and increased the ratio of cholesterol in high density lipoprotein (HDL) to the cholesterol in LDL + VLDL. Although a 10% cod liver oil addition also reduced the total cholesterol level, the ratio of HDL/LDL + VLDL was similar to that of the 10% butter-cholesterol diet. (4) A direct relationship was found between the concentration of oleic acid (18:1) in serum and the total cholesterol level (r = 0.947) and also the level of LDL + VLDL-cholesterol (r = 0.935). These results show that cod liver oil, wheat germ oil, and phytosterol induce an increase in the PUFA/SFA ratio, promote hypocholesterolemia, and change lipoprotein concentration. However, there were indications that no relationship exists between the change in the total cholesterol level and the change in the ratio of HDL/LDL + VLDL, and that the increase of total cholesterol and LDL + VLDL-cholesterol was consistent with the increase of oleic acid in serum.

An imbalance between formation and detoxification of oxygen radicals leads to oxidant stress that may increase in more intense oxidative metabolism caused by a high intake of metabolizable energy to provide metabolic intermediates for the milk synthesis and secretion. This hypothesis was tested using dairy cows and the concentration of hydroperoxides in lipids (LHP) extracted from circulative lipoprotein particles of low and very low density (LDL and VLDL/chylomicrons) as oxidant stress indicator. The particles were prepared by ultracentrifugation of serum obtained by coccygeal bleeding (13 cows, 1. parity, n=8 and 2. parity, n=5, lactation stage, 53 +/- 1.4 days post partum) and purified by precipitation. Concentrations of LHP-LDL/mg Lipoprotein correlated significantly with daily milk yield (r = 0.73, P = 0.004) or daily milk energy output (r = 0.77, P = 0.003) in contrast to LHP of VLDL/chylomicron particles. Thus, some evidence was obtained for an almost linear, positive relationship between milk productivity and oxidant stress occurring in LDL.

In the total fraction of low and very low density lipoproteins (LDL+VLDL) isolated from serum by precipitation in the presence of heparin-Mn the copper-induced lipid peroxidation was accompanied by accumulation of LPO products, a decrease ANS fluorescence intensity (F(ANS)) and an increase in probe--cation DSP-6, a fluorescence intensity decrease of intrinsic in the ultraviolet area (F(uv)) and an increase in the visible area (F(vis)). The degree of lipoprotein modification was estimated by calculating the F(vis)/F(uv) and F(DSP)-6/F(ANS) ratio. Strong positive correlation was found between these ratios and concentration of thiobarbituric acid-reactive substances (TBARS) of LDL+VLDL samples isolated from sera of 49 donors and incubated at 37 degrees C in the presence of 50 M CuSO4 during 0, 3 and 24 hr (F(vis)/F(uv) (r = 0.75; p < 0.001) and F(DSP-6)/F(ANS) (r = 0.73; p < 0.001)). Very strong positive correlation was also found between both fluorescent parameters F(vis)/F(uv) and F(DSP-6)/F(ANS) (r = 0.95, p < 0.001). Changes in the values of F(vis)/ F(uv), F(DSP-6)/F(ANS), concentration of TBARS in 75 patients with documented coronary heart disease (CHD) and 49 apparently healthy donors were studied. No significant differences of these parameters in LDL+VLDL of patients with CHD and donors were found.

We have previously demonstrated that monatepil maleate, AJ-2615, a new calcium antagonist endowed with alpha1-adrenoceptor blocking property, has antiatherosclerotic and plasma lipid-lowering effects in Japanese monkeys fed on a cholesterol-rich diet. To clarify the mechanisms on plasma lipid-lowering action, we investigated the effect of monatepil maleate in these monkeys on hepatic acyl-CoA:cholesterol acyltransferase (ACAT) activity. Both ACAT activity and esterified cholesterol content in the livers of monkeys fed on a cholesterol-rich diet for 6 months significantly increased about 7- and 16-fold, respectively, as compared with those in monkeys fed on a standard diet. Monatepil maleate (30 mg/kg/day for 6 months, orally) inhibited the increases of ACAT activity and esterified cholesterol content by 51% and 71%, respectively. In in vitro experiments, monatepil maleate inhibited ACAT activity in a concentration-dependent manner, whereas it did not affect 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase activity. A kinetic analysis revealed that monatepil maleate was a noncompetitive type inhibitor of ACAT. Hepatic ACAT activity was significantly correlated to hepatic esterified cholesterol content (r = 0.775, P < .0001), to plasma very low density lipoprotein (VLDL) content (r = 0.765, P < .0001) and to plasma total cholesterol content (r = 0.573, P < .005) in the monkeys. These results suggest that ACAT-inhibiting effect of monatepil maleate plays an important role in the reduction of hyperlipidemia.

We describe the development and performance of a homogeneous assay for the direct turbidimetric determination of LDL particles in human serum. The assay is based upon the specific agglutination of LDL particles by the polyanion PAMPS. The co-agglutination of VLDL is avoided by the addition of a zwitterionic detergent. Yielding results within 10 min, the assay requires only a small sample volume taken directly from primary serum tubes, i.e., no pretreatment of the sample is necessary. It can be easily applied to routine clinical chemistry analyzers. The results are highly correlated with LDL cholesterol determinations by ultracentrifugation (r)0.95) and dextran sulfate precipitation (r)0.95), but an increased recovery of small, high density LDL particles is observed, which more adequately reflects the atherogenic risk of LDL. The assay provides excellent intra- and inter-assay CVs in the range of 0.6--1.6 and 1.7--2.4%, respectively, on Roche Diagnostics/Hitachi analyzers. The method is well suited to the high-throughput screening of LDL cholesterol levels.

Equine plasma lipoproteins were fractionated into VLDL, LDL-1, LDL-2 and HDL by density gradient ultracentrifugation. From each lipoprotein fraction, five apo C like peptides of approx. M(r) 1400, 10000, 9500, 9000 and 8000 were detected by SDS-polyacrylamide gel electrophoresis. After partial purification by Sephadex G-75, one fraction, showing a strong activation of lipoprotein lipase, was further purified by Mono Q anion exchange column. Two of the apo C like peptides (M(r) 10000 and 8000) activated the bovine milk lipoprotein lipase in vitro; only one (M(r) 9500) inhibited the lipolytic activity. This work confirms that many mammals present two apo C-II components with different molecular weights.

Rats were kept either on a standard laboratory diet or a high cholesterol, olive oil diet for periods ranging from 1 day to 22 weeks. The effect of the high cholesterol, olive oil diet on the concentrations of cholesterol, glycosaminoglycans (GAGs) and collagen in aortic intima-media, were studied and the developing hyperlipidemia was characterized. The concentration of cholesterol in rat aorta was increased after 22 weeks' high cholesterol, olive oil diet, while collagen concentration was not affected. On the contrary, the concentration of aortic sulphated GAGs was significantly increased already after one week's high cholesterol, olive oil diet. The diet increased the formation of a cholesterol-rich very low density lipoprotein (VLDL), decreased high density lipoprotein (HDL)-associated cholesterol and phospholipids, but had virtually no effect on low density lipoprotein (LDL)-lipids. The concentrations of VLDL-cholesterol and -phospholipids showed positive correlations with the concentration of aortic GAGs (r = 0.89 and 0.83, respectively, P less than 0.05 for both). Stronger (negative) correlations were found between aortic GAGs and HDL-cholesterol and -phospholipids (r = 0.94 for both, P less than 0.01) suggesting that HDL may have a role in the control of arterial sulphated GAG concentration.

<Abst<em>r</em>actText>obesity is a ch<em>r</em>onic disease associated with inadequate eating habits and <em>r</em>educed levels of physical activity. Because of obesity, the <em>r</em>isk fo<em>r</em> como<em>r</em>bidities is inc<em>r</em>eased, especially fo<em>r</em> ca<em>r</em>diovascula<em>r</em> diseases, insulin <em>r</em>esistance, and inc<em>r</em>eased p<em>r</em>o-inflammato<em>r</em>y facto<em>r</em>s. The aim of the p<em>r</em>esent investigation was to analyze potential co<em>r</em><em>r</em>elations between p<em>r</em>o/anti-inflammato<em>r</em>y adipokines, glycemic index, and othe<em>r</em> ma<em>r</em>ke<em>r</em>s of diet quality using a metabolic p<em>r</em>ofile in women unde<em>r</em>going inte<em>r</em>disciplina<em>r</em>y weight loss the<em>r</em>apy.</Abst<em>r</em>actText><Abst<em>r</em>actText>thi<em>r</em>ty-two women with obesity we<em>r</em>e en<em>r</em>olled in a 12-week p<em>r</em>og<em>r</em>am of inte<em>r</em>disciplina<em>r</em>y the<em>r</em>apy combining a clinical, nut<em>r</em>itional, and physical exe<em>r</em>cise app<em>r</em>oach. Body composition, quality of diet, metabolic p<em>r</em>ofile, and p<em>r</em>o/anti-inflammato<em>r</em>y adipokines we<em>r</em>e analyzed.</Abst<em>r</em>actText><Abst<em>r</em>actText>the the<em>r</em>apy showed to be effective in <em>r</em>educing body weight, body mass index, and body fat. The<em>r</em>e was also an imp<em>r</em>ovement in lipid p<em>r</em>ofile, including total choleste<em>r</em>ol, non-HDL-choleste<em>r</em>ol, <em>VLDL</em>-choleste<em>r</em>ol, t<em>r</em>iglyce<em>r</em>ides and glucose metabolism, including glucose, and insulin. As fo<em>r</em> food intake, the<em>r</em>e was a dec<em>r</em>ease in calo<em>r</em>ie consumption (f<em>r</em>om 1991.45 ± 677.78 to 1468.88 ± 390.56; p = 0.002), ca<em>r</em>bohyd<em>r</em>ates (f<em>r</em>om 50.37 ± 6 to 47.04 ± 8.67; p = 0.04), lipids (f<em>r</em>om 31.83 ± 5.53 to 30.37 ± 7.04; p = 0.3), and glycemic load (f<em>r</em>om 80.53 ± 39.88 to 54.79 ± 23.69; p = 0.02), and an inc<em>r</em>eased consumption of p<em>r</em>oteins (f<em>r</em>om 18.3 ± 2.39 to 22.89 ± 4.9; p = 0.002). Positive co<em>r</em><em>r</em>elations we<em>r</em>e demonst<em>r</em>ated between insulin concent<em>r</em>ation and waist ci<em>r</em>cumfe<em>r</em>ence (<em>r</em> = 0.82; p = 0.003); leptin and body fat and abdominal ci<em>r</em>cumfe<em>r</em>ence (<em>r</em> = 0.74; p = 0.01); and LDL-choleste<em>r</em>ol f<em>r</em>action and total choleste<em>r</em>ol consumption (<em>r</em> = 0.69; p = 0.027). Negative co<em>r</em><em>r</em>elations we<em>r</em>e demonst<em>r</em>ated between leptin and monosatu<em>r</em>ated fat consumption (<em>r</em> = -0.71; p = 0.02); and adiponectin and live<em>r</em> enzyme GGT levels (<em>r</em> = -0.65; p = 0.04).</Abst<em>r</em>actText><Abst<em>r</em>actText>inte<em>r</em>disciplina<em>r</em>y the<em>r</em>apy had positive effects on inflammato<em>r</em>y state, mediated by leptin, adiponectin, and quality of diet. Ou<em>r</em> findings suggest the effectiveness and clinical <em>r</em>elevance of the inte<em>r</em>disciplina<em>r</em>y clinical the<em>r</em>apy applied fo<em>r</em> obesity.</Abst<em>r</em>actText>

We examined whether lipid metabolism in orchiectomised (ORX) rats was affected by fructose ingestion and the amount of ingested fructose. Sucrose was used as a fructose source. Sham-operated and ORX rats were fed one of the following three diets for 28 d: a maize starch-based diet without sucrose (SU0), a diet by which half or all of maize starch was replaced by sucrose (SU50 or SU100). Body-weight gain and food intake were increased by sucrose ingestion, but decreased by ORX. Plasma total cholesterol concentration was increased by ORX and dose-dependently by sucrose ingestion. Plasma TAG concentration was decreased by ORX, but was increased dose-dependently by sucrose ingestion. Plasma insulin concentration was decreased by ORX, but was not affected by sucrose ingestion. Liver TAG was increased by sucrose ingestion and ORX; however, liver cholesterol concentration was not affected by sucrose ingestion and ORX. The hepatic activity of cholesterol 7α-hydroxylase 1 was not affected by sucrose ingestion and ORX; however, faecal excretion of bile acids was decreased. The mRNA level of microsomal TAG transfer protein, which is the gene related to hepatic VLDL production, was increased by ORX and sucrose ingestion. The mRNA level of uncoupling protein-1 was decreased by ORX, but not by sucrose ingestion. Plasma insulin concentration tended to correlate with the level of sterol-regulatory element-binding protein-1c mRNA (r 0·747, P = 0·088). These results show that lipid metabolism in ORX rats would be affected by the consumption of fructose-rich sweeteners such as sucrose and high-fructose syrup.

BACKGROUND

the prevalence of obesity has increased, especially among women.

OBJECTIVE

the aim of this study was to assess the effect of a hypoenergetic diet combined with coconut flour on anthropometric and biochemical data and the quality of the diet.

METHODS

we carried out a crossover clinical trial involving a step with hypoenergetic diet only and another with the diet associated with coconut flour consumption (26 g) over the course of nine months. The volunteers were recruited from the São Gonçalo city of Rio de Janeiro. Anthropometric, biochemical and dietary data were collected monthly. The diet quality index revised for the Brazilian population (DQI-R) and the consumption of ultra-processed foods and additives were assessed. The Wilcoxon and Mann-Whitney tests were performed, with p < 0.05 considered statistically significant.

RESULTS

forty-two women of an average 47.5 ± 9.5 years of age participated. The hypoenergetic diet promoted a decrease in body fat, body mass index, waist circumference, waist-to-height ratio, visceral adiposity index, diastolic blood pressure, triglycerides and VLDL. The consumption of coconut flour promoted a drop in glucose and total cholesterol levels when supplementing the hypoenergetic diet. The improvement to diet quality can be noted in the decrease in consumption of ultra-processed foods like vegetable oil, chocolate and soft drinks.

CONCLUSIONS

the hypoenergetic diet promoted a decrease in the anthropometric parameters, blood pressure and triglycerides. The consumption of coconut flour promoted a decrease in glucose and total cholesterol levels when supplementing the hypoenergetic diet. The improved diet quality can be seen in the decrease in consumption of ultra- processed foods.

We compared two HPLC methods (anion exchange [AE] and steric exclusion [SE]) for analysis of mouse lipoprotein profiles by determining coefficients of variability (CVs) under varying conditions. CVs for AE and SE were comparable on fresh samples. There was an inverse relationship between subfraction curve area and CV [r = -0.65 (AE) and -0.50 (SE)], consistent with the interpretation that as curve area decreased, error variance increased and signal-to-noise ratio decreased. Sample storage did not affect SE. In contrast, with AE, alterations in measured lipoproteins were apparent after storage, including a decrease in the HDL subfraction [66.8% (baseline) vs. 15.9% (1 week); P < 0.01] and an increase in areas under LDL and VLDL peaks. Concomitant with decreasing HDL area, reproducibility deteriorated with the duration of storage. Analysis of the effects of decreasing sample injectate volume to <25 microl on SE lipoprotein subfractions revealed that areas under LDL and VLDL peaks decreased and persisted as volume was decreased further. Areas under all lipoprotein subfractions measured with either AE or SE were linearly correlated with the amount of cholesterol [r = 0.69 (AE) and 0.87 (SE)]. Both AE and SE yield reproducible, accurate, and rapid measurements of lipoproteins from small amounts of serum. AE yields more sensitive, high-amplitude, well-defined peaks that can be easily distinguished and necessitates the use of smaller sample volumes compared with SE, but sample storage causes alterations in the chromatogram. SE appears better suited to serial analyses involving stored samples.

<Abst<em>r</em>actText>Ch<em>r</em>onic kidney disease (CKD) associates with complex lipop<em>r</em>otein distu<em>r</em>bances <em>r</em>esulting in high ca<em>r</em>diovascula<em>r</em> <em>r</em>isk. Apolipop<em>r</em>otein E (APOE) is a polymo<em>r</em>phic p<em>r</em>otein with th<em>r</em>ee common isofo<em>r</em>ms (E2; E3; E4) that plays a c<em>r</em>ucial <em>r</em>ole in lipop<em>r</em>otein metabolism, including hepatic clea<em>r</em>ance of chylomic<em>r</em>ons and ve<em>r</em>y low-density lipop<em>r</em>otein (<em>VLDL</em>) <em>r</em>emnants, and <em>r</em>eve<em>r</em>se choleste<em>r</em>ol t<em>r</em>anspo<em>r</em>t. It demonst<em>r</em>ates anti-athe<em>r</em>ogenic p<em>r</em>ope<em>r</em>ties but data conce<em>r</em>ning the link between polymo<em>r</em>phism and level of APOE in CKD patients a<em>r</em>e inconclusive. The aim of ou<em>r</em> <em>r</em>esea<em>r</em>ch was to assess the <em>r</em>elationship between APOE gene polymo<em>r</em>phism and APOE concent<em>r</em>ation and its <em>r</em>edist<em>r</em>ibution among lipop<em>r</em>oteins along with CKD p<em>r</em>og<em>r</em>ession.</Abst<em>r</em>actText><Abst<em>r</em>actText>90 non-dialysed CKD patients we<em>r</em>e included into the study. Real time PCR was used fo<em>r</em> APOE genotyping. APOE level was measu<em>r</em>ed in se<em>r</em>um and in isolated lipop<em>r</em>otein f<em>r</em>actions (<em>VLDL</em>; IDL + HDL; HDL). Kidney function was assessed using eGFR CKD-EPI fo<em>r</em>mula.</Abst<em>r</em>actText><Abst<em>r</em>actText>The population was divided into th<em>r</em>ee APOE genotype subg<em>r</em>oups: E2(ε2ε3), E3(ε3ε3) and E4(ε3ε4). The highest APOE level was obse<em>r</em>ved fo<em>r</em> the E2 subg<em>r</em>oup (p < 0.001). APOE concent<em>r</em>ation positively co<em>r</em><em>r</em>elated with eGFR value in the E2 subg<em>r</em>oup (<em>r</em> = 0.7, p < 0.001) but inve<em>r</em>sely in the E3 subg<em>r</em>oup (<em>r</em> = - 0.29, p = 0.02).). A lowe<em>r</em> concent<em>r</em>ation of APOE in the E2 subg<em>r</em>oup was associated with its diminished contents in HDL and IDL + LDL pa<em>r</em>ticles. In the E3 subg<em>r</em>oup, the highe<em>r</em> concent<em>r</em>ation of APOE was <em>r</em>elated to the inc<em>r</em>eased numbe<em>r</em> of non-HDL lipop<em>r</em>oteins.</Abst<em>r</em>actText><Abst<em>r</em>actText>In patients with CKD, APOE genotype as well as <em>r</em>enal function a<em>r</em>e associated with the concent<em>r</em>ation of APOE and its <em>r</em>edist<em>r</em>ibution among lipop<em>r</em>otein classes.</Abst<em>r</em>actText>

OBJECTIVE

To evaluate the association between very low density lipoprotein cholesterol (VLDL-C) and cholesterol absorption and synthesis markers in patients with moderate and high risk of coronary heart disease.

METHODS

A total 363 statin-naïve patients with moderate and high risk of coronary heart disease were consecutively recruited from two hospitals in Shanxi and Henan provinces between October 2008 and June 2009. A standard questionnaire and physical examination were performed at baseline. Atorvastatin (20 mg/day) was administered to patients for 4 weeks. Venous blood samples after an overnight fast were collected before and after treatment for measuring VLDL-C and cholesterol absorption and synthesis markers. In qualitative analyses, the baseline level of cholesterol absorption and synthesis markers and their reduction after atorvastatin treatment were categorized into 3 tertile groups.

RESULTS

(1) Of 363 patients, 283 patients with mean age of (55.43±9.01)years old with complete data were finally analyzed. The median level of baseline VLDL-C was 1.06 (0.65, 1.86) mmol/L. The median level of baseline cholesterol absorption marker (Campesterol) and cholesterol synthesis marker (Lathosterol) was 6.01 (3.78, 9.45) mg/L and 13.46 (8.30, 21.07) mg/L, respectively. (2) Partial correlation analysis and multiple regression showed the baseline level of VLDL-C was positively correlated with Campesterol (r=0.153, P<0.05) but not with Lathosterol(r=0.182, P=0.173). Furthermore, baseline VLDL-C level significantly increased with tertile of the baseline level of Campesterol in the qualitative analyses(P for trend=0.035). (3) Mean reduction in VLDL-C levels was 38.0% after 4 weeks atorvastatin treatment. VLDL-C reduction was positively correlated with Campesterol reduction (r=0.331, P<0.001). VLDL-C reduction significantly increased with the tertile of Campesterol reduction (P for trend=0.032). But this trend was not observed between VLDL-C level and Lathosterol (P for trend=0.798).

CONCLUSIONS

The level of VLDL-C was closely related to cholesterol absorption marker, and further studies are needed to validate if inhibitor of cholesterol absorption (for example by Ezetimibe) could bring about more effective VLDL-C lowering effect in this patient cohort.

OBJECTIVE

South Asians have higher rates of type 2 diabetes and cardiovascular disease compared to most other racial/ethnic groups. Increased hepatic de novo lipogenesis (DNL) in response to dietary sugar may accelerate the development of these chronic diseases in this population.

METHODS

Hepatic DNL in response to a calorically sweetened beverage was measured in an outpatient setting in 15 South Asians and 15 Caucasians with similar and normal body mass indexes, waist circumferences, glucose tolerance and lipid profiles. Blood was sampled before and hourly for 4 h after the ingestion of a single beverage made with glucose (1·5 g/kg) and fructose (1·5 g/kg). The main outcome, DNL, was measured as the increase in %palmitate (16:0) in very low-density lipoprotein (VLDL) triglyceride (TG) over 4 h.

RESULTS

After the sugar dose, the increase in %16:0 in VLDL TG was significantly greater in South Asians vs Caucasians (P = 0·01). VLDL and total TG also increased to a significantly greater extent in South Asians (P = 0·04 and <0·001, respectively). Although the fasting and postsugar levels of insulin and glucose did not differ between groups, the DNL response significantly correlated with the insulin response to sugar in South Asians (r = 0·56, P = 0·03).

CONCLUSIONS

Hepatic DNL in response to a sugar challenge was greater in healthy, young South Asians compared to Caucasians despite normal indices of insulin sensitivity, and it correlated with the insulin response. These findings suggest an early, insulin-related, gene-nutrient interaction contributing to the high prevalence of diabetes and coronary disease in this population.

Sepsis is a life-threatening organ dysfunction syndrome caused by a dysregulated host response to infection. Proprotein convertase subtilisin/kexin type 9 (PCSK9) is often upregulated in the presence of sepsis and infectious diseases. In sepsis, PCSK9 degraded the low-density lipoprotein cholesterol (LDL) receptors (LDL-R) of the hepatocytes and the very low-density lipoprotein cholesterol receptors (VLDL-R) of the adipocytes, which then subsequently reduced pathogenic lipid uptake and clearance/sequestration. Moreover, it might improve cholesterol accumulation and augment toll-like receptor function in macrophages, which supported inflammatory responses. Accordingly, PCSK9 might show detrimental effects on immune host response and survival in sepsis. However, the exact roles of PCSK9 in the pathogenesis of sepsis are still not well defined. In this review, we summarized the literatures focusing on the roles of PCSK9 in sepsis. Our review provided an additional insight in the role of PCSK9 in sepsis, which might serve as a potential target for the treatment of sepsis.

To study the hyoplipidemic activity of non-methoxy and methoxy substituted--alkyl indan acetic acids in normogenic and hypergenic animal models. The hyoplipidemic activity was evaluated using normogenic and hypergenic rat models. Indan acids synthesized in our laboratory (50 mg/kg) and the standard drug clofibrate (50 mg/kg) were used for this study. Dimethoxy substituted -methyl and -ethyl indan acetic acids (9 & 10) exhibited significant hypocholesterolemic activity in both animal models. Clofibrate showed cholesterol lowering activity of 18% in normogenic rats while test agents 9 and 10 produced a similar activity of 20% and 17% respectively. Liver cholesterol and liver weight of the animals tested were found normal in case of the test compounds while liver weight was increased by 15% in clofibrate treated rats. LDL and VLDL cholesterol levels were also affected by compounds No. 9 and 10. Compounds having a smaller alkyl groups (R = CH3, C2H5) at--carbon atom of the acetic acid moiety exhibited significant hypocholesterolemic activity.

Apolipoprotein E concentrations in plasma were investigated in 17 patients with hepatocellular (non-cholestatic) liver disease. Ninety-five per cent of plasma apolipoprotein E was recovered in density fractions less than 1.063 kg/l and it was therefore associated with very low-density lipoproteins (VLDL) and low-density lipoproteins (LDL). Very low-density lipoproteins and LDL apolipoprotein E correlated positively (r = 0.63, p less than 0.01) with plasma bile salt concentrations when a decrease in liver protein synthesis was taken into account. This may be due to a down-regulation of the hepatic apolipoprotein B, E receptors concomitant with increasing plasma bile salt concentrations.

To explore the intracellular signal pathways for beta-VLDL induced very low density lipoprotein receptor (VLDL-R) transcription up-regulation and their effects on lipid accumulation in macrophages, Western Blot was used to examine phosphorylated ERK1/2 protein and regulated effects by different singal kinase inhibitants. It was found that beta-VLDL induced an increase in ERK1/2 activity in a protein kinase C (PKC)-dependent manner in murine RAW264.7 macrophages. By using different protein kinases inhibitors or activators, it was observed that the effect of beta-VLDL induced VLDL receptor transcription, which was monitored by RT-PCR analysis of VLDL receptor mRNA, was not affected by the inhibitor of p38 kinase and cAMP analog, but extremely abolished by pretreating cells with PD98059, an inhibitor of ERK and GF 109203X, an inhibitor of PKC. These results demonstrated that the PKC-ERK1/2 cascade is the essential signaling pathway by which beta-VLDL activated VLDL-R mRNA expression. Inhibition of the ERK1/2 signaling cascade resulted in suppression of the cellular lipid accumulation induced by beta-VLDL in macrophages.

We describe a new method, useful to clinical laboratories, for assessing intermediate density (IDL) or beta-very-low-density (beta-VLDL) lipoprotein cholesterol. The technique involves selective precipitation properties of the qualitative Wieland and Seidel post-electrophoretic method that immobilizes IDL and beta-VLDL in the beta-zone of an agarose slide (Clin Chem 1973;19:1139-41). In our method, we separate low-density lipoprotein (LDL) in a second electrophoretic step, in which LDL moves toward the anode, and then quantify the cholesterol of the above lipoproteins remaining in the precipitate band at the beta-zone. Replicate within-run precision (CV) of 15 aliquots of a sera pool was 10.1%. The correlation with sequential ultracentrifugation of 30 samples was r = 0.96 (P less than 0.001). Serum reference values for 30 normal individuals are 57 +/- 7.0 mg/L. Seven phenotype III hyperlipoproteinemic patients had the highest concentrations of IDL or beta-VLDL cholesterol in serum, 1620 +/- 346 mg/L.

The ligand-binding domain of VLDL receptor contains eight imperfectly similar repeats. To discuss the contribution of each repeat to ligand binding, the RT-PCR technique was used to clone the VLDLR-cDNA from the heart muscle of Chinese people. Two recombinants were further constructed, which contained the full-length cDNA of VLDLR and the mutant lacking repeats 1-5. CHO cell line was transfected with two recombinants. The expression of VLDLR gene could be detected by RT-PCR from the CHO cells transfected with pCD-VR. The results of binding experiments showed that the ability of the CHO cells transfected with the full-length cDNA of VLDL-R binding DiI-labeled beta-VLDL was higher than that of the CHO cells transfected with the mutant. Our findings indicated that human VLDL-R gene could be expressed effectively on CHO cells, and the receptor was almost inactivated when repeats 1-5 were deleted.

Breast cancer as one of the most prevalent cancers has high morbidity and mortality. Very low-density lipoprotein receptor (VLDLR) is a multifunctional receptor which plays a principal role in the tumor development through affecting cell metastasis and proliferation. The VLDLR as a target for miRNA-4465 and miRNA-1297 was predicted using bioinformatics analysis. Tissue specimens of malignant (n = 50), benign (n = 35) and corresponding normal breast (n = 20) were considered to evaluate the expression of VLDLR using RT-qPCR and western blotting. The VLDL cholesterol (VLDL-C) levels were quantified using a colorimetric assay. The relative VLDLR expression was found in the malignant tumors, which was significantly lower than that in the normal tissues (P<0.05). The expression levels of VLDLR had no significant difference between malignant and benign tissues (P>0.05). Correlation analysis revealed that the VLDLR expression level had a direct correlation with miRNA-1297 (R=0.566, P<0.05), but a reverse one with miRNA-4465 (R = -0.663, P<0.0001). The VLDL-C level in the malignant and normal tissues was lower than that in the benign tumors, which was not significant (P>0.05). The expression levels of VLDLR in E⁺P-H^- (ER⁺,PR^-,HER-) tumors were higher than those in other subtypes (P<0.05). Furthermore, a negative correlation was found between the VLDLR expression level and the Ki 67% score. These data revealed that the lower expression of VLDLR mediated by the high expression levels of miRNA-4465 may be involved in the development of breast cancer. These results might provide some evidence for the effect of VLDLR on the breast cancer.

Keywords: Benign; Breast cancer; Malignant; RT-qPCR; VLDLR; Western blotting.