Cholesterol‐imprinted polymer receptor prepared by a hybrid imprinting method

A novel cholesterol‐imprinted polymer (CMIP‐H) was prepared by a hybrid method of covalent imprinting and non‐covalent imprinting. This approach involves the copolymerization of a template‐containing monomer, cholesteryl 2‐hydroxyethyl methacrylate carbonate, and a cross‐linker, followed by hydrolysis to afford a flexible guest‐binding site accompanied with the easy and efficient removal of a ‘sacrificial spacer’. The effect of solvent on the binding capacity of CMIP‐H towards cholesterol was studied, indicating that a good binding capacity towards cholesterol could be achieved in a less‐polar solvent. The binding experiments of CMIP‐H towards a series of structural analogues of cholesterol, including cholesterol acetate, progesterone and stigmasterol, were carried out in hexane. The results showed that CMIP‐H almost did not bind cholesterol acetate at all because the hydrogen‐bonding site is blocked. It exhibited a similar binding towards both cholesterol and stigmasterol, but much higher binding towards progesterone. Copyright © 2005 Society of Chemical Industry     

Simultaneous HPLC quantification of total cholesterol,tocopherols and β-carotene in Barrosã-PDO veal

A simple, rapid and sensitive procedure for the simultaneous determination of total cholesterol, tocopherols and β-carotene in meat is described. The method involves a direct saponification of the meat, a single n-hexane extraction and the analysis of the extracted compounds by normal-phase HPLC, using fluorescence (tocopherols) and UV–Vis photodiode array (cholesterol and β-carotene) detections in tandem. Rates of recovery of spiked meat samples were 93% for cholesterol, 83–86% for (α-, β- and -γ) tocopherols and 89% for β-carotene. Repeatabilities were high (CV < 6%) for all determined compounds, except for δ-tocopherol. This tocopherol, which is not usually present in meat, showed a much lower recovery percentage (73%) and repeatability (12.8%). This methodology was applied for the quantification of total cholesterol, tocopherols and β-carotene in three muscles (longissimus thoracis, longissimus lumborum and semitendinosus) of the Portuguese traditional Barrosã-PDO veal, obtained from autochthonous calves fed extensively during summer (with the least abundant green pastures) and slaughtered in early autumn (October). Barrosã-PDO veal showed median contents of total cholesterol (0.50–0.56 mg/g) and, depending on the analysed muscle, moderate to high contents of α-tocopherol (3.3–3.9 μg/g) and β-carotene (0.07–0.09 μg/g), suggesting an high sensorial and hygienic quality.     

TNFα-Induced LDL Cholesterol Accumulation Involve Elevated LDLR Cell Surface Levels and SR-B1 Downregulation in Human Arterial Endothelial Cells

Excess lipid droplets are frequently observed in arterial endothelial cells at sites of advanced atherosclerotic plaques. Here, the role of tumor necrosis factor alpha (TNFα) in modulating the low-density lipoprotein (LDL) content in confluent primary human aortic endothelial cells (pHAECs) was investigated. TNFα promoted an up to 2 folds increase in cellular cholesterol, which was resistant to ACAT inhibition. The cholesterol increase was associated with increased ¹²⁵I-LDL surface binding. Using the non-hydrolysable label, Dil, TNFα could induce a massive increase in Dil-LDL by over 200 folds. The elevated intracellular Dil-LDL was blocked with excess unlabeled LDL and PCSK9, but not oxidized LDL (oxLDL), or apolipoprotein (apoE) depletion. Moreover, the TNFα-induced increase of LDL-derived lipids was elevated through lysosome inhibition. Using specific LDLR antibody, the Dil-LDL accumulation was reduced by over 99%. The effects of TNFα included an LDLR cell surface increase of 138%, and very large increases in ICAM-1 total and surface proteins, respectively. In contrast, that of scavenger receptor B1 (SR-B1) was reduced. Additionally, LDLR antibody bound rapidly in TNFα-treated cells by about 30 folds, inducing a migrating shift in the LDLR protein. The effect of TNFα on Dil-LDL accumulation was inhibited by the antioxidant tetramethythiourea (TMTU) dose-dependently, but not by inhibitors against NF-κB, stress kinases, ASK1, JNK, p38, or apoptosis caspases. Grown on Transwell inserts, TNFα did not enhance apical to basolateral LDL cholesterol or Dil release. It is concluded that TNFα promotes LDLR functions through combined increase at the cell surface and SR-B1 downregulation.     

Evaluation of Floxuridine Oligonucleotide Conjugates Carrying Potential Enhancers of Cellular Uptake

Conjugation of small molecules such as lipids or receptor ligands to anti-cancer drugs has been used to improve their pharmacological properties. In this work, we studied the biological effects of several small-molecule enhancers into a short oligonucleotide made of five floxuridine units. Specifically, we studied adding cholesterol, palmitic acid, polyethyleneglycol (PEG 1000), folic acid and triantennary N-acetylgalactosamine (GalNAc) as potential enhancers of cellular uptake. As expected, all these molecules increased the internalization efficiency with different degrees depending on the cell line. The conjugates showed antiproliferative activity due to their metabolic activation by nuclease degradation generating floxuridine monophosphate. The cytotoxicity and apoptosis assays showed an increase in the anti-cancer activity of the conjugates related to the floxuridine oligomer, but this effect did not correlate with the internalization results. Palmitic and folic acid conjugates provide the highest antiproliferative activity without having the highest internalization results. On the contrary, cholesterol oligomers that were the best-internalized oligomers had poor antiproliferative activity, even worse than the unmodified floxuridine oligomer. Especially relevant is the effect induced by palmitic and folic acid derivatives generating the most active drugs. These results are of special interest for delivering other therapeutic oligonucleotides.     

Thai Tea Seed Oil and Virgin Olive Oil Similarly Reduce Plasma Lipids: A Pilot Study within a Healthy Adult Male Population

The potential cardiovascular benefit of virgin olive oil (VOO) is widely recognized. However, the use of VOO at very high cooking temperatures makes these oils poorly suited for many Asian dishes. The use of tea seed oil (TSO) is increasing in Thailand, with TSO having a higher smoke point than VOO. The current study examines the effects of daily TSO intake in healthy adults. In a randomized, single-blind crossover design, 12 men consumed for 3 weeks 40 g day⁻¹ of food prepared with either TSO or VOO as a cooking oil. Plasma lipids, thiobarbituric acid reactive substances (TBARS), and oxidant defense enzyme activities are measured before and after each 3-week intervention period. Gas chromatography analysis of TSO and VOO demonstrates that both oils are equally high in monounsaturated fatty acid. The dietary incorporation of TSO and VOO for three weeks reduces low-density lipoprotein cholesterol (LDL-C) concentrations by 15% and 13%, respectively; with total cholesterol (TC) levels lowered by 10% in both groups. No significant changes in TBARS or antioxidant enzyme activity is observed. These results support the concept that Thai TSO can be utilized as a suitable and healthy alternative oil for high-temperature cooking in many Thai and Asian diets. Practical Applications: Tea seed oil from Camellia oleifera grown in Thailand has been recently reported to favorably lower lipid profiles in hamsters fed a high-fat diet in a manner similar to feeding refined olive oil or grapeseed oil. A pilot crossover trial is conducted to compare the effects of three weeks of daily intake of either TSO or VOO in healthy human adults. Consumption of both oils produced significant reductions in TC and LDL-C. Thai TSO leads to favorable lipid profiles and is a reasonable choice for many Thai and Asian food recipes.     

胆固醇氧化酶高产菌株的诱变选育及产酶条件优化

在对细脚拟青霉、玫烟色拟青霉、粉拟青霉、香菇、平菇、金针菇等胆固醇氧化酶产生菌初筛的基础上．选择以产酶较高的玫烟色拟青霉作为紫外诱变的出发菌株,筛选出一株相对高产突变菌株MFEC006,其酶活达到了0．5483U／mL,比出发菌株提高了154％,经8次传代培养,突变株性质稳定．对MFEC006菌株产酶条件进行优化,得出其最佳产酶条件为：pH为6．5,温度27℃,接种量10％,摇床转速180r／min．     

A Cross-sectional Study on the Relationship Between Homocysteine and Lipid Profiles Among Chinese Population from Hunan

Previous studies have explored the relationship between homocystein (Hcy) and lipid profiles. However, the results from these studies have been inconsistent. The current study investigated the correlation between Hcy and lipid profiles in Chinese community-based population. The participants were composed of 4012 Chinese people aged 30–92 years old, who were recruited from rural and urban communities in the Hunan Province. Non-parametric test and logistic regression were used to examine the distribution of Hcy and lipid profiles (triglyceride [TG], total cholesterol [TC], low-density lipoprotein cholesterol [LDL-C], high-density lipoprotein cholesterol [HDL-C]) and the relationship between them. The median age of subjects was 54.50 years old, and 40.98% were male. Median Hcy was 13.20 μmol/L, and 35.39% had hyperhomocysteinemia (HHcy). Median TG was 1.51 mmol/L, TC was 4.77 mmol/L, LDL-C was 2.62 mmol/L, and HDL-C was 1.27 mmol/L. In multivariable logistic regression analysis, HHcy was associated with high levels of TG (OR_male = 2.240, p < 0.001; OR_female = 2.539, p < 0.001), TC (OR_male = 2.237, p < 0.001; OR_female = 2.202, p < 0.001), and LDL-C (OR_male = 1.413, p = 0.010; OR_female = 1.617, p < 0.001) in the different sexes population and low level of HDL-C in females (OR = 1.326, p = 0.023) after adjusting for confounders. HHcy was independently associated with an increasing risk of low HDL-C among females. The regression analysis showed that HHcy was also associated with hypertriglyceridemia, hypercholesterolemia, and high level of LDL-C in males and females from Chinese community-based population, which provides a basis for the treatment and prevention of abnormal lipid metabolism.     

Association between Single Nucleotide Polymorphism rs1044925 and the Risk of Coronary Artery Disease and Ischemic Stroke

The present study was performed to clarify the association between the acyl-CoA:cholesterol acyltransferase-1 (ACAT-1) single nucleotide polymorphism (SNP) rs1044925 and the risk of coronary artery disease (CAD) and ischemic stroke (IS) in the Guangxi Han population. Polymerase chain reaction and restriction fragment length polymorphism was performed to determine the genotypes of the ACAT-1 SNP rs1044925 in 1730 unrelated subjects (CAD, 587; IS, 555; and healthy controls; 588). The genotypic and allelic frequencies of rs1044925 were significantly different between the CAD patients and controls (p = 0.015) and borderline different between the IS patients and controls (p = 0.05). The AC/CC genotypes and C allele were associated with a decreased risk of CAD and IS (CAD: p = 0.014 for AC/CC vs. AA, p = 0.022 for C vs. A; IS: p = 0.014 for AC/CC vs. AA; p = 0.017 for C vs. A). The AC/CC genotypes in the healthy controls, but not in CAD or IS patients, were associated with an increased serum high-density lipoprotein cholesterol (HDL-C) concentration. The present study shows that the C allele carriers of ACAT-1 rs1044925 were associated with an increased serum HDL-C level in the healthy controls and decreased risk in CAD and IS patients.