Lipoprotein Subfraction Profile and HDL-Associated Enzymes in Sickle Cell Disease Patients

Although hypocholesterolemia is a reported finding in sickle cell disease (SCD), low-density lipoprotein (LDL)/high-density lipoprotein (HDL) subfractions and HDL-associated enzymes have not been determined in SCD patients. Blood was collected from 38 hemoglobin (Hb)A volunteers and 45 homozygous HbSS patients who had not received blood transfusions in the last 3 months. Serum lipids were measured by automated analyzer while LDL and HDL subfraction analysis was done by continuous disc polyacrylamide gel electrophoresis. Serum levels of cholesteryl ester transfer protein (CETP), lecithin-cholesterol acyltransferase (LCAT), apolipoprotein B (apoB) and apolipoprotein A-1 (apoA-I) were determined by enzyme-linked immunosorbent assay (ELISA). Total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) levels were significantly decreased, while TG levels were significantly increased in SCD patients compared to controls. A significant decrease in intermediate-density lipoprotein (IDL)-C, IDL-B, IDL-A and LDL-1 fractions were seen in SCD patients, while no significant difference was observed in small dense LDL particles. A significant decrease was seen in HDL-large, HDL-intermediate and HDL-small fractions in SCD patients versus controls. Levels of LCAT and ApoA-1 protein measured in SCD patients were significantly lower while no significant difference was observed in CETP and ApoB protein levels compared to controls. The reduction observed in LDL- and HDL-C in SCD patients was reflected as significantly decreased IDL, LDL-1 and HDL-subfractions. Decreased HDL subfractions may possibly lead to the reduced ApoA-1 and LCAT protein levels observed in SCD patients.     

三鹿降血脂奶粉调节血脂作用的研究

观察了三鹿降血脂奶粉调节血脂的作用。用Wistar品系雄性大鼠60只随机分为6组,全部饲以高脂饲料,在此基础上分别饲以蒸馏水、普通奶粉及3个不同剂量的降血脂奶粉。实验期45d,实验前后测量体重,并采用酶法测定血清总胆固醇(TC)、总甘油三酯(TG)及高密度脂蛋白胆固醇(HDL-C)。结果表明,实验前后的各组大鼠体重均明显增加,但各实验组间无显著性差异(P>0.05)。处理组大鼠血清TG水平显著低于对照组(P<0.05或P<0.01);高剂量降血脂奶粉组大鼠血清HDL-C水平显著高于对照组(P<0.05);各组大鼠血清TC水平则未见有显著性差异。结论,三鹿降血脂奶粉对实验大鼠体重无影响,但可以显著降低TG并可显著升高HDL-C水平。     

降血脂鼠李糖乳杆菌的筛选及其对高脂模型地鼠血脂的影响

该实验旨在筛选出一株具有较强降血脂功能的乳杆菌。试验采用的菌株分离自健康婴儿肠道,通过邻苯二甲醛比色法测定菌株降胆固醇能力、单试剂法测定降甘油三酯能力,并测定了其对酸及胆盐的耐受性,从中筛选出具有降胆固醇和降甘油三酯功能较强的菌株进行16S rDNA鉴定。通过建立高血脂地鼠模型,研究菌株体内降血脂能力。结果表明,鼠李糖乳杆菌HCS01-013的降胆固醇和甘油三酯能力较强,其胆固醇降解率达到35.40%;甘油三酯降解率达到44.32%;在0.3%和1.5%的胆盐环境下培养17 h的存活率分别为99.67%和96.79%,在pH 3.0和pH 2.0环境下培养17 h后存活率能达到88.76%和71.34%;黄金地鼠喂养高脂饲料建立的高脂地鼠模型后,喂养鼠李糖乳杆菌HCS01-013各剂量组与喂养生理盐水的模型对照组相比甘油三酯含量均明显降低。鼠李糖乳杆菌HCS01-013具有一定的降血脂功能,且耐逆环境能力较强,为该菌株相关研究及产品研发提供理论依据。     

Oat-based Symbiotic Beverage Fermented by Lactobacillus plantarum, Lactobacillus paracasei ssp. casei, and Lactobacillus acidophilus

ABSTRACT: Oats and probiotics have long been recognized for their health benefits. The objectives of this study were (1) to study the ability of Lactobacillus plantarum (B-28), Lactobacillus paracasei ssp. casei (B-29) isolated from a traditional Bulgarian cereal-based fermented beverage, and Lactobacillus acidophilus from Chr. Hansen, Milwaukee, Wis., U.S.A., to remove cholesterol from the media and to adhere to the Caco-2 cell line, (2) to optimize the fermentation conditions to develop a beverage using these probiotics and oats with acceptable sensory and nutritional qualities, and (3) to assess the quality and shelf-life of this beverage and survivability of probiotics in the beverage. Lactobacillus acidophilus , B-28, and B-29 were found to remove 70.67%± 2.35%, 20.26%± 2.63%, and 16.75%± 3.83% of cholesterol from media and the percentage of adhesion was 4.69%± 0.78%, 1.92%± 0.78%, and 8.36%± 0.78%, respectively. The blend of oat flour, sugar, inulin, and whey protein concentrate was cooked in water and fermented for 12 h at 37°C by 2 ± 10⁶ colony-forming units (CFU) /mL each of B-28 and B-29 and 2 ± 10⁸ CFU/mL of L. acidophilus . The beverage had 0.87%± 0.03% of dietary fiber and had better sensory qualities compared with the commercially available similar product. The probiotics survived for 10 wk of storage at 4°C, except for L. acidophilus , which survived for about 4 wk. The population of B-28 was 1.77 ± 10⁶ to 1.29 ± 10⁷ CFU/mL and that of B-29 was 7.39 ± 10⁷ to 4.49 ± 10⁸ CFU/mL throughout the storage period. The oat-based symbiotic beverage is a functional drink providing both probiotics and prebiotics at the same time.     

Heat-moisture treatment of high-amylose corn starch increases dietary fiber content and lowers plasma cholesterol in ovariectomized rats

ABSTRACT:  The effect of dietary high-amylose corn starch (HACS) of varying dietary fiber (DF) content on plasma cholesterol was examined in ovariectomized (OVX) rats. Gelatinized normal corn starch (G-CS) was used as a reference. OVX rats were fed a fiber-free purified diet containing G-CS, HACS, gelatinized high-amylose corn starch (G-HACS), or heat-moisture treated HACS (HM-HACS) at 400 g starch/kg diet for 21 d. The DF content of G-CS, HACS, G-HACS, and HM-HACS measured by the AOAC method was 0.1, 19.3, 2.4, and 64.5 g/100 g, respectively. The dry weight of cecal contents, cecal wall weight, the amount of short chain fatty acids in cecal contents, the amount of bile acids in small intestinal contents, and fecal excretion of neutral sterols increased logarithmically with increasing DF, while total plasma cholesterol concentration decreased. On the other hand, hepatic CYP7A1 activity, fecal dry weight, and fecal excretion of bile acids increased linearly with increasing DF, while body weight gain decreased. The hypocholesterolemic effect of HACS in OVX-rats appeared to be more effective by heat-moisture treatment.     

Immobilization of cholesterol oxidase and potassium ferricyanide on dodecylbenzene sulfonate ion‐doped polypyrrole film

Dodecylbenzene sulfonate (DBS)‐doped polypyrrole (PPY) conducting polymer films were electrochemically deposited onto the indium‐tin‐oxide (ITO)‐coated glass plates in aqueous medium. The enzyme cholesterol oxidase (ChOx) was immobilized on these DBS–PPY films by a physical adsorption technique. These ChOx‐immobilized DBS–PPY films were characterized by ultraviolet–visible and Fourier transform infrared spectroscopy. The enzyme activity studies indicate that ～40% of ChOx leaches out from the ChOx/DBS–PPY film. The ChOx activity in the ChOx/DBS–PPY film was assayed as a function of cholesterol concentration. The results of amperometric measurements conducted on ChOx/DBS–PPY/ITO film show linearity over the range 2–8 mM of cholesterol solution. The ChOx/DBS–PPY/ITO electrodes exhibit a response time of 30 s and are stable for ～3 months at 4 °C. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 82: 3486–3491, 2001     

2,4-二氨基苯基胆甾醚的合成

以溴苯为原料,经硝化后,在四丁基溴化铵相转移催化剂作用下与胆固醇成醚、硝基还原反应得到2,4-二氨基苯基胆甾醚,并对成醚反应的条件进行了优化。较佳合成反应条件是:以甲苯为溶剂,n(2,4-二硝基溴苯)∶n(胆固醇)=1.55∶1,反应温度25℃,w(NaOH)=50%,反应时间3.5 h,产率为72%。所得中间体和目标化合物经过红外光谱和核磁波谱的表征。     

Effect of Repetitive Lysine-Tryptophan Motifs on the Eukaryotic Membrane

In a previous study, we synthesized a series of peptides containing simple sequence repeats, (KW)_n–NH₂ (n = 2,3,4 and 5) and determined their antimicrobial and hemolytic activities, as well as their mechanism of antimicrobial action. However, (KW)₅ showed undesirable cytotoxicity against RBC cells. In order to identify the mechanisms behind the hemolytic and cytotoxic activities of (KW)₅, we measured the ability of these peptides to induce aggregation of liposomes. In addition, their binding and permeation activities were assessed by Trp fluorescence, calcein leakage and circular dichrorism using artificial phospholipids that mimic eukaryotic liposomes, including phosphatidylcholine (PC), PC/sphingomyelin (SM) (2:1, w/w) and PC/cholesterol (CH) (2:1, w/w). Experiments confirmed that only (KW)₅ induced aggregation of all liposomes; it formed much larger aggregates with PC:CH (2:1, w/w) than with PC or PC:SM (2:1, w/w). Longer peptide (KW)₅, but not (KW)₃ or (KW)₄, strongly bound and partially inserted into PC:CH compared to PC or PC:SM (2:1, w/w). Calcein release experiments showed that (KW)₅ induced calcein leakage from the eukaryotic membrane. Greater calcein leakage was induced by (KW)₅ from PC:CH than from PC:SM (2:1, w/w) or PC, whereas (KW)₄ did not induce calcein leakage from any of the liposomes. Circular dichroism measurements indicated that (KW)₅ showed higher conformational transition compared to (KW)₄ due to peptide-liposome interactions. Taken together, our results suggest that (KW)₅ reasonably mediates the aggregation and permeabilization of eukaryotic membranes, which could in turn explain why (KW)₅ displays efficient hemolytic activity.     

The Association of Cholesterol Uptake and Synthesis with Histology and Genotype in Cortisol-Producing Adenoma (CPA)

Cortisol-producing adenoma (CPA) is composed of clear and compact cells. Clear cells are lipid abundant, and compact ones lipid poor but associated with higher production of steroid hormones. PRKACA mutation (PRKACA mt) in CPA patients was reported to be associated with more pronounced clinical manifestation of Cushing’s syndrome. In this study, we examined the association of histological features and genotypes with cholesterol uptake receptors and synthetic enzymes in 40 CPA cases, and with the quantitative results obtained by gas chromatography-mass spectrometry (GC-MS) analysis in 33 cases to explore their biological and clinical significance. Both cholesterol uptake receptors and synthetic enzymes were more abundant in compact cells. GC-MS analysis demonstrated that the percentage of compact cells was inversely correlated with the concentrations of cholesterol and cholesterol esters, and positively with the activity of cholesterol biosynthesis from cholesterol esters. In addition, hormone-sensitive lipase (HSL), which catalyzes cholesterol biosynthesis from cholesterol esters, tended to be more abundant in compact cells of PRKACA mt CPAs. These results demonstrated that both cholesterol uptake and biosynthesis were more pronounced in compact cells in CPA. In addition, more pronounced HSL expression in compact cells of PRKACA mt CPA could contribute to their more pronounced clinical manifestation.