| For medical practitioners and the general public - Cholesterol Journal Article Catalog. |
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| Gallbladder contractility in aspirin- and cholesterol-fed prairie dogs Li, Y. F., D. H. Russell, et al. (1994), Gastroenterology 106(6): 1662-7. Abstract: BACKGROUND/AIMS: Whether aspirin prevents cholesterol gallstone formation is controversial. This study aimed to investigate this issue and determine the depression of gallbladder smooth muscle contractility associated with cholesterol feeding in the prairie dog. METHODS: Prairie dogs were divided into four subgroups. Animals were fed control or 1.2% cholesterol diet and treated with placebo or aspirin for 2 weeks. The presence of crystals and stones was determined, and contractile force in response to cholecystokinin octapeptide (CCK-8) of gallbladder muscle strips was measured. RESULTS: Maximal stress of 2.66 +/- 0.23 x 10(4) N/m2 was measured in muscle strips from animals on control diet. Maximal stress was significantly lower in strips from animals on high-cholesterol diet, being 1.49 +/- 0.16 x 10(4) N/m2 with placebo and 1.62 +/- 0.23 x 10(4) N/m2 with aspirin. The difference in maximal stress between aspirin-treated and placebo-treated animals was not significant. Although none of the animals on control diet had crystals or stones, all animals on the high-cholesterol diet, whether receiving placebo or aspirin, had crystals in the bile, and more than 65% had cholesterol stones. CONCLUSIONS: Aspirin has no effect on stone formation, nor does it prevent the decrease in contractility despite a profound decrease in endogenous gallbladder prostanoid synthesis. |
| Gallbladder emptying in vivo, bile composition, and nucleation of cholesterol crystals in patients with cholesterol gallstones Stolk, M. F., K. J. van Erpecum, et al. (1995), Gastroenterology 108(6): 1882-8. Abstract: BACKGROUND/AIMS: Impaired postprandial gallbladder emptying may provide time for progressive bile concentration with formation of instable cholesterol-rich vesicles and fast nucleation of cholesterol crystals. The aim of this study was to assess postprandial gallbladder emptying, bile composition, and nucleation of cholesterol crystals in the same patient. METHODS: In 30 patients with cholesterol gallstones, postprandial gallbladder emptying was measured ultrasonographically. In each patient, gallbladder bile composition (obtained at cholecystectomy) and nucleation of cholesterol crystals was determined. Patients were divided in 22 strong contractors (> 50% postprandial gallbladder emptying) and 8 weak contractors. RESULTS: In weak contractors, bile salt and phospholipid concentrations were much higher than in strong contractors (234.6 +/- 24.7 vs. 130.3 +/- 10.8 mmol/L P < 0.001 and 44.5 +/- 3.5 vs. 30.2 +/- 3.1 mmol/L P < 0.05, respectively). Cholesterol concentrations were comparable in strong and weak contractors. Consequently, total lipid concentration was significantly higher (15.5 +/- 1.4 and 9.2 +/- 0.7 g/dL; P < 0.001) and cholesterol saturation index significantly lower (0.90 +/- 0.08 and 1.61 +/- 0.17; P < 0.001) in weak contractors. Nucleation time, percentage of cholesterol in vesicles, bile salt species, and molecular species of phosphatidylcholine were not significantly different. CONCLUSION: Differences in bile composition can be linked to different patterns of postprandial gallbladder emptying and may point to two different pathways of gallstone formation. |
| Gallbladder motility and cholesterol crystallization in bile from patients with pigment and cholesterol gallstones Portincasa, P., A. Di Ciaula, et al. (2000), Eur J Clin Invest 30(4): 317-24. Abstract: BACKGROUND: Little is known about gallbladder motility in patients with black pigment stones when compared to cholesterol gallstone patients, or about their relationship to biliary composition, crystallization and stone characteristics. DESIGN: Fasting and postprandial gallbladder volumes were studied by ultrasonography in 49 gallstone patients with pigment (n = 14) or cholesterol (n = 35) stones and 30 healthy controls. After cholecystectomy stone composition, gallbladder wall inflammation, cholesterol saturation index and appearance of platelike cholesterol crystals in bile were evaluated in gallstone patients. RESULTS: Fasting gallbladder volume was significantly (P < 0.05) increased in cholesterol stone patients (31.7 +/- 1.9 mL) but not in pigment stone patients (21.9 +/- 3.1 mL), compared to controls (21.0 +/- 1.5 mL). Postprandial emptying was delayed in patients (half-emptying time: 31 +/- 2 min, 35 +/- 3 min, 24 +/- 2 min in cholesterol stone patients, pigment stone patients and controls, respectively, P < 0.05) and incomplete (residual volume: 43.2 +/- 2.7%, 40.0 +/- 4.3%, 15.8 +/- 1.6% min in cholesterol stone patients, pigment stone patients and controls, respectively, P < 0.05). The inflammation of the gallbladder wall was mild or absent in all cases. Biliary cholesterol saturation index was 152.3 +/- 8.5% and 92.9 +/- 4.8% in patients with cholesterol and pigment stones, respectively (P < 0.01). Whereas cholesterol crystals never appeared during 21 days in biles from patients with pigment stones, crystal observation time in patients with cholesterol gallstone was 5 days (median) and was significantly shorter in patients with multiple (4 days) than in patients with solitary (12 days) cholesterol stones (P = 0.0019). CONCLUSIONS: Patients with black pigment stones who do not have excess cholesterol and do not grow cholesterol crystals in bile have decreased gallbladder emptying, although to a lesser extent than patients with cholesterol stones. Thus, gallbladder stasis is likely to put a subset of subjects at risk for the formation of pigment gallstones, and pathogenic mechanisms need to be further investigated. |
| Gallbladder motility in cholesterol gallstone disease. Effect of ursodeoxycholic acid administration and gallstone dissolution Festi, D., R. Frabboni, et al. (1990), Gastroenterology 99(6): 1779-85. Abstract: Gallbladder motility was evaluated by ultrasonography in 75 cholesterol gallstone patients and in 77 matched control subjects. All 75 gallstone patients were candidates for oral bile acid therapy (radiolucent gallstones, less than 2 cm in diameter, in well-opacified gallbladder), and 38 of them were also studied during ursodeoxycholic acid administration. An additional 20 gallstone patients were studied 1 year after confirmed gallstone dissolution with oral bile acids. Gallstone patients showed significantly greater fasting and residual volumes, a decreased percent of gallbladder emptying, but a similar absolute emptying and emptying rate compared with the control subjects. Greater fasting volumes and reduced percents of gallbladder emptying were also found in gallstone-free patients who achieved complete dissolution with oral bile acids. After ursodeoxycholic acid administration, fasting gallbladder volumes were greater, and percents of gallbladder emptying were further decreased than in untreated gallstone patients. In conclusion, greater fasting volumes, and not reduced gallbladder contractility, account for the defective gallbladder function in radiolucent (cholesterol-rich) gallstone patients. This condition is likely to precede, and possibly to promote, gallstone formation because it persists after gallstone dissolution. Ursodeoxycholic acid administration worsens the defect observed in gallstone patients. This finding also suggests, although indirectly, that the expected normalization of cholesterol saturation during oral bile acid administration is not paralleled by an improvement in gallbladder function. |
| Gallbladder motor function, plasma cholecystokinin and cholecystokinin receptor of gallbladder in cholesterol stone patients Zhu, J., T. Q. Han, et al. (2005), World J Gastroenterol 11(11): 1685-9. Abstract: AIM: To study the interactive relationship of gallbladder motor function, plasma cholecystokinin (CCK) and cholecystokinin A receptor (CCK-R) of gallbladder in patients with cholesterol stone disease. METHODS: Gallbladder motility was studied by ultrasonography in 33 patients with gallbladder stone and 10 health subjects as controls. Plasma CCK concentration was measured by radioimmunoassay in fasting status (CCK-f) and in 30 min after lipid test meal (CCK-30). Radioligand method was employed to analyze the amount and activity of CCK-R from 33 gallstone patients having cholecystectomy and 8 persons without gallstone died of severe trauma as controls. RESULTS: The percentage of cholesterol in the gallstone composition was more than 70%. The cholesterol stone type was indicated for the patients with gallbladder stone in this study. Based on the criterion of gallbladder residual fraction of the control group, 33 gallstone patients were divided into two subgroups, contractor group (14 cases) and non-contractor group (19 cases). The concentration of CCK-30 was significantly higher in non-contractor group than that in both contractor group and control group (55.86+/-3.86 pmol/L vs 37.85+/-0.88 pmol/L and 37.95+/-0.74 pmol/L, P<0.01), but there was no difference between contractor group and control group. Meanwhile no significant difference of the concentration of CCK-f could be observed among three groups. The amount of CCK-R was lower in non-contractor group than those in both control group and contractor group (10.27+/-0.94 fmol/mg vs 24.59+/-2.39 fmol/mg and 22.66+/-0.55 fmol/mg, P<0.01). The activity of CCK-R shown as KD in non-contractor group decreased compared to that in control group and contractor group. Only was the activity of CCK-R lower in contractor group than that in control group. The ejection fraction correlated closely with the amount of CCK-R (r = 0.9683, P<0.01), and the concentration of CCK-30 correlated negatively with the amount of CCK-R closely (r = -0.9627, P<0.01). CONCLUSION: The distinctive interactive relationship of gallbladder emptying, plasma CCK and CCK-R in gallbladder from this study suggested that the defect of CCK-R may be a key point leading to the impairment of gallbladder motor function and the pathogenesis of cholesterol gallstone formation may differ in two subgroups of gallstone patient, gallbladder non-contractor group or contractor group. |
| Gallbladder mucin as a pronucleating agent for cholesterol monohydrate crystals in bile Smith, B. F. (1990), Hepatology 12(3 Pt 2): 183S-186S; discussion 186S-188S. Abstract: Mucin is a densely glycosylated macromolecule secreted by the gallbladder epithelium as the principal constituent of gallbladder mucus. Hypersecretion of gallbladder mucus occurs in response to a lithogenic diet in experimental animals, and mucus accumulates as a viscous gel within the gallbladder lumen before gallstone formation. In both animals and man, the initial stage of cholesterol gallstone formation, the nucleation of cholesterol monohydrate crystals, occurs within the mucus gel. Inhibition of mucus secretion with aspirin prevents gallstone formation in the cholesterol-fed prairie dog, indicating the importance of mucus in gallstone formation. Mucin contains domains that bind cholesterol and lecithin transported as vesicles in supersaturated bile. Furthermore, mucin accelerates the nucleation of cholesterol crystals in both supersaturated model and native biles. Binding of cholesterol-enriched vesicles to hydrophobic domains on the mucin protein core appears to be critical for the acceleration of cholesterol crystal nucleation by mucin. Further study of the structure and function of gallbladder mucin should help to elucidate the pathogenesis of cholesterol cholelithiasis. |
| Gallbladder mucosal blood flow increases during early cholesterol gallstone formation Conter, R. L., J. L. Washington, et al. (1992), Gastroenterology 102(5): 1764-70. Abstract: Gallbladder absorption increases during early cholesterol gallstone formation and is influenced by the intraluminal presence of lithogenic bile. The effect of lithogenic bile on gallbladder mucosal blood flow is unknown. The current study tested the hypothesis that the presence of lithogenic gallbladder and hepatic bile enhances gallbladder mucosal blood flow in cholesterol-fed (0.4%) prairie dogs, as determined by hydrogen gas clearance. Gallbladder mucosal blood flow in control animals was 35.57 +/- 3.9 mL.min-1.100 g-1. In contrast, basal gallbladder mucosal blood flow in cholesterol-fed animals was significantly (P less than 0.01) increased to 64.94 +/- 8.7 mL.min-1.100 g-1. In crossover studies, the addition of lithogenic gallbladder bile to control animals (n = 6) resulted in a significant (P less than 0.025) 26% increase in gallbladder mucosal blood flow, whereas the addition of nonlithogenic gallbladder bile into gallbladders of cholesterol-fed prairie dogs resulted in a significant (P less than 0.025) 58% decrease in gallbladder mucosal blood flow. Similarly, hepatic bile crossover studies showed that the addition of lithogenic hepatic bile to control gallbladders significantly increased (P less than 0.025) gallbladder blood flow by 30%, whereas instillation of nonlithogenic hepatic bile in gallbladders of cholesterol-fed animals significantly (P less than 0.025) decreased gallbladder mucosal blood flow by 29%. These results suggest that alterations in gallbladder mucosal blood flow, influenced by the presence and absence of lithogenic bile, may play a role in cholesterol gallstone formation. |
| Gallbladder relaxation in patients with pigment and cholesterol stones Chen, Q., J. Amaral, et al. (1997), Gastroenterology 113(3): 930-7. Abstract: BACKGROUND & AIMS: Gallbladders with cholesterol stones show a defective contraction in response to agonists. The aim of this study was to investigate the muscle relaxation of human gallbladders with cholesterol or black pigment gallstones. METHODS: Gallbladder relaxation was measured in vitro using muscle strips and single muscle cells. Relaxation was expressed as percent inhibition of either basal active tension in strips or maximal cell contraction induced by diacylglycerol. The production of cyclic nucleotides was determined using a 125I-labeled radioimmunoassay kit. RESULTS: Frequency-dependent relaxation evoked by electrical field stimulation was significantly lower in gallbladders with cholesterol stones than in gallbladders with pigment stones. Relaxation and adenosine 3',5'-cyclic monophosphate (cAMP) production induced by isoproterenol, vasoactive intestinal peptide, and forskolin were also significantly decreased in gallbladders with cholesterol stones. However, the relaxation in response to 8-bromo-cAMP, nitric oxide (NO), and the NO donor S-nitroso-N-acetylpenicillamine (SNAP), which circumvent plasma membrane receptors and directly activate intracellular mechanisms, was similar in gallbladders with cholesterol and pigment stones. Guanosine 3',5'-cyclic monophosphate production induced by NO and SNAP was also similar. CONCLUSIONS: Human gallbladder muscle from specimens with cholesterol stones show an impaired relaxation and lower cAMP production compared with specimens with pigment stones. The muscle defect(s) responsible for this impairment seem to be in the plasma membranes. |
| Gallbladder stones classified by chemical analysis of cholesterol content. Frederiksberg, 1987-1988 Ravnborg, L., D. Teilum, et al. (1990), Scand J Gastroenterol 25(7): 720-4. Abstract: Gallstones from 80 cholecystectomies and 81 autopsies were chemically analysed and showed a trend of decreasing cholesterol content with increasing age (p = 0.00009). The frequency of cholesterol stones (cholesterol content greater than 70%) was higher in operated women (81%) than in operated men (33%) (p = 0.0006) and in the total autopsy material (42%). The study supports the theory that the predominance of gallstone disease in women is an effect of the preponderance of cholesterol stones. The accuracy of estimates of cholesterol content of gallstones from the appearance of the cut surface was low. When the chemical analysis was used as a key, only one in three was correct. There was a tendency to underestimate cholesterol content. |
| Galloyl esters from rhubarb are potent inhibitors of squalene epoxidase, a key enzyme in cholesterol biosynthesis Abe, I., T. Seki, et al. (2000), Planta Med 66(8): 753-6. Abstract: Galloyl glucoses and galloyl proanthocyanidins obtained from rhubarb (Rhei Rhizoma, Rheum palmatum L., Polygonaceae); e.g. 1,2,6-tri-O-galloyl-beta-D-glucose (IC50 = 0.63 microM), 1,6-di-O-galloyl-2-O-cinnamoyl-beta-D-glucose (IC50 = 0.58 microM), procyanidin B-2 3,3'-di-O-gallate (IC50 = 0.54 microM), and procyanidin B-5 3,3'-di-O-gallate (IC50 = 0.55 microM), were found to be potent inhibitors of rat squalene epoxidase (SE). The inhibition at submicromolar level was far more potent than that of chemically synthesized substrate analogs. It was demonstrated for the first time that the cholesterol-lowering effect of rhubarb may be attributed to the potent inhibition activities of SE, a rate-limiting enzyme of cholesterol biogenesis. |
| Gallstone cholesterol content is related to apolipoprotein E polymorphism Juvonen, T., K. Kervinen, et al. (1993), Gastroenterology 104(6): 1806-13. Abstract: BACKGROUND: The genetically determined phenotypes of apolipoprotein E are related to variations in lipoprotein levels and in the enterohepatic metabolism of cholesterol and bile acids. The present study was designed to elucidate the role of apolipoprotein E polymorphism in gallstone formation. METHODS: Apolipoprotein E phenotype was determined in 169 consecutive cholecystectomy patients and in 200 controls. The cholesterol content of the gallstones (n = 169), the presence of cholesterol monohydrate crystals of fresh gallbladder bile (n = 142), and the nucleation time (n = 35) were also analyzed. RESULTS: The median cholesterol content of the gallstones was higher in the apolipoprotein E4 category (phenotypes E4/4 and E4/3, 97%) than in the E3 (E3/3, 78%) and E2 patients (E2/2 and E2/3, 76%, P = 0.0003). In E4 patients, cholesterol crystals were found immediately after surgery in 27 of 40 (68%), whereas in E3 and E2 groups in 36 of 88 (41%), and 4 of 14 (29%) of the patients (P = 0.0001). The median nucleation time in E4 patients (2.5 days) was shorter than in patients with E3 (5.5 days) or E2 (6.0 days) (P = 0.0016). CONCLUSIONS: These data indicate that apolipoprotein E polymorphism affects cholesterol content of cholelithiasis. We suggest that this phenomenon is mediated by the altered formation of cholesterol monohydrate crystals in different apolipoprotein E phenotypes. |
| Gallstones in patients with morbid obesity. Relationship to body weight, weight loss and gallbladder bile cholesterol solubility Shiffman, M. L., H. J. Sugerman, et al. (1993), Int J Obes Relat Metab Disord 17(3): 153-8. Abstract: Gallstones are common in obesity, and in individuals undergoing weight reduction. However, the relationships between body weight, weight reduction, gallbladder bile composition and gallstone formation are not well understood. The present studies were conducted on a cohort of 230 morbidly obese individuals presenting for bariatric surgery. Mean body weight ranged from 90-235.4 kg (mean: 136.2 kg). Body mass index (BMI) ranged from 35.4-94.7 kg/m2. Thirty-two patients (14%) had undergone prior cholecystectomy and 48 (21%) were found to have gallstones by intraoperative ultrasonography. No significant relationship was observed between gallstone prevalence and body weight. Following bariatric surgery weight loss averaged 1.57 kg/week over six months. Absolute weight loss ranged from 13.6-81.3 kg. Symptomatic gallstones requiring cholecystectomy developed in 15/150 patients (10%) over two years of follow-up. In contrast, ultrasonography detected asymptomatic gallstones in 34/92 patients (37%) six months following bariatric surgery. No relationship existed between the amount of weight lost and gallstone formation. Gallbladder bile cholesterol solubility remained constant throughout the entire weight range present in this population. No significant difference in cholesterol solubility was present between persons presenting for bariatric surgery and patients who developed symptomatic gallstones and underwent cholecystectomy following weight reduction. We conclude that gallstones are common in patients with severe obesity both before and following bariatric surgery. However, weight loss per se does not appear to be the major determinant of gallstone formation in persons who weigh in excess of 100 kg. |
| Gamma-glutamyl transpeptidase and acetylcholinesterase activities in brain capillaries of cholesterol-fed rabbits Eryurek, F. G., E. Surmen, et al. (1990), Res Commun Chem Pathol Pharmacol 69(2): 245-8. Abstract: Cerebral microvascular endothelium, the constituent cell of the blood-brain barrier, is enriched in the enzyme gamma-glutamyl transpeptidase (GGT). This enzyme plays a role in the regulation of amino acid uptake and transport, and in the gamma-glutamylation of serotonin, dopamine and norepinephrine. Recent studies have demonstrated that GGT activity is modulated by cholinergic-adrenergic agonists. The levels of the acetylcholine-catabolizing enzyme acetylcholinesterase (AChE), may therefore be related to the modulation of the GGT activity. In this study, the activities of GGT and AChE in microvessel-enriched fractions were assayed after feeding of rabbits a high cholesterol diet. A 21% decrease of GGT activity and a 44% increase of AChE activity appeared at the end of dietary treatment. |
| Gamma-secretase activity is present in rafts but is not cholesterol-dependent Wada, S., M. Morishima-Kawashima, et al. (2003), Biochemistry 42(47): 13977-86. Abstract: Cholesterol has been claimed to be involved in the generation and/or accumulation of amyloid beta protein (Abeta). However, the underlying molecular mechanisms have not been fully elucidated yet. Here, we have investigated the effect of membrane cholesterol content on gamma-secretase activity using Chinese hamster ovary cells stably expressing beta-amyloid precursor protein (APP) and either wild-type or N141I mutant-type presenilin 2. Cholesterol was acutely depleted from the isolated membrane by methyl-beta-cyclodextrin, and Abeta production was assessed in a cell-free assay system. Reduced cholesterol did not significantly alter the amounts of Abeta produced by either total cell membranes or cholesterol-rich low-density membrane domains. Even its extremely low levels in the latter domains did not affect Abeta production. This indicates that the membrane cholesterol content does not directly modulate the activity of gamma-secretase. To ascertain that gamma-secretase resides in cholesterol-rich membrane domains, low-density membrane domains were further fractionated with BCtheta (biotinylated theta-toxin nicked with subtilisin Carlsberg protease), which has recently been shown to bind selectively to rafts of intact cells. The membrane domains purified with BCtheta did indeed produce Abeta. These observations indicate that the gamma-cleavage required for generating Abeta occurs in rafts, but its activity is virtually cholesterol-independent. |
| Garlic reduces plasma lipids by inhibiting hepatic cholesterol and triacylglycerol synthesis Yeh, Y. Y. and S. M. Yeh (1994), Lipids 29(3): 189-93. Abstract: Prompted by the reported hypolipidemic activity of garlic, the present study was undertaken to elucidate the mechanism(s) underlying the cholesterol-lowering effects of garlic. Rat hepatocytes in primary culture were used to determine the short-term effects of garlic preparations on 1-14Cacetate and 2-3Hglycerol incorporation into cholesterol, fatty acids and glycerol lipids. When compared with the control group, cells treated with a high concentration of garlic extracts i.e., petroleum ether- (PEF), methanol- (MEF) and water-extractable (WEF) fractions from fresh garlic showed decreased rates of 1-14Cacetate incorporation into cholesterol (by 37-64%) and into fatty acids (by 28-64%). Kyolic containing S-allyl cysteine and organosulfur compounds inhibited cholesterogenesis in a concentration dependent manner with a maximum inhibition of 87% at 0.4 mM. At this concentration, Kyolic decreased 1-14Cacetate incorporation into fatty acids by 67%. S-allyl cysteine at 2.0 and 4.0 mM inhibited cholesterogenesis by 20-25%. PEF, MEF and WEF depressed the rates of 2-3Hglycerol incorporation into triacylglycerol, diacylglycerol and phospholipids in the presence of acetate, but not in the presence of oleate. The results suggest that the hypocholesterolemic effect of garlic stems, in part, from decreased hepatic cholesterogenesis, whereas the triacylglycerol-lowering effect appears to be due to inhibition of fatty acid synthesis. Primary hepatocyte cultures as used in the present study have been proven useful as tools for screening the anticholesterogenic properties of garlic principles. |
| Gas chromatographic determination of cholesterol and tocopherols in edible oils and fats with automatic removal of interfering triglycerides Ballesteros, E., M. Gallego, et al. (1996), J Chromatogr A 719(1): 221-7. Abstract: An automated gas chromatographic method for the simultaneous determination of cholesterol, alpha-tocopherol and alpha-tocopheryl acetate in edible oils and fats without derivatization is reported. Interferences from lipid material are avoided by using a continuous system to transesterify triglycerides with potassium methylate in methanol. The precision of the method is 1.9, 2.2 and 3.1% for cholesterol, alpha-tocopherol and alpha-tocopheryl acetate, respectively. The proposed methods was validated by analysing a standard reference material of coconut oil (SRM 1563-2) with good results. The method features a high throughput, minimal sample handling and analyte specificity (lipid material does not interfere). |
| Gas chromatographic determination of phospholipid fatty acids and free cholesterol in a single sample Briuzgina, T. S., E. Kravchenko, et al. (1991), Lab Delo(9): 18-20. Abstract: The results of gas chromatographic measurements of phospholipid fatty acid spectrum and free cholesterol in the blood serum and red cells of diabetics are presented. A deficit of polyunsaturated fatty acids (at the expense of C18:2 and C20:4) in the presence of a high free cholesterol level was found to be a characteristic feature of diabetes mellitus. |
| Gas chromatographic properties of common cholesterol and phytosterol oxidation products Apprich, S. and F. Ulberth (2004), J Chromatogr A 1055(1-2): 169-76. Abstract: The most common cholesterol and phytosterol oxidation products found in foodstuffs or biological matrices are the 7alpha- and 7beta-hydroxysterol, 7-ketosterol, 5alpha,6alpha- and 5beta,6beta-epoxysterol, and triol derivatives of sterols. This study focused on the preparation and purification of such products derived from campesterol, stigmasterol and beta-sitosterol. The identity of the substances was confirmed by mass spectroscopic analysis. The elution order of a complex mixture composed of the 7alpha- and 7beta-hydroxysterol, 7-ketosterol, 5alpha,6alpha- and 5beta,6beta-epoxysterol, and triol derivatives of cholesterol, campesterol, stigmasterol and beta-sitosterol was recorded on an apolar as well as a medium-polarity capillary column in relation to two commonly used internal standards, i.e. alpha-cholestane and 19-hydroxy cholesterol. Flame-ionization detector as well as mass spectrometry response factors were derived from a gravimetrically prepared mixture of commercially available cholesterol oxide standards. It was proven that the ionization efficiency of cholesterol and phytosterol oxides are very similar and that response factors obtained for cholesterol oxidation products are also valid for quantitative work regarding phytosterol oxidation products. |
| Gas chromatography-mass spectrometric method for quantitative determination in human urine of dicarboxylic (dioic) acids produced in the body as a consequence of cholesterol biosynthesis inhibition Jemal, M. and Z. Ouyang (1998), J Chromatogr B Biomed Sci Appl 709(2): 233-41. Abstract: A capillary gas chromatography-mass spectrometric (GC-MS) method in human urine has been developed and validated for the quantitative determination of dicarboxylic acids (dioic acids) which are produced in the body as a consequence of the administration of an inhibitor of the enzyme squalene synthase, which is involved in the biosynthesis of cholesterol. The standards and quality control (QC) samples were prepared by adding dioic acids into human urine. Internal standard (sebacic acid) was added to each urine sample (0.1 ml) and then dried by evaporation under nitrogen. The dried sample was reacted with pentafluorobenzyl (PFB) bromide under conditions that maximized the formation of the di-PFB ester (at the expense of the mono-PFB ester) of the dioic acids. After drying by evaporation, each sample residue was reconstituted in mesitylene and injected into a capillary GC-MS system via a splitless injection. The detection was by negative ion chemical ionization mass spectrometry with selected ion monitoring (SIM) of the M-PFB- of the analytes and the internal standard. |
| Gastric helicobacter infection induces a Th2 phenotype but does not elevate serum cholesterol in mice lacking inducible nitric oxide synthase Ihrig, M., M. T. Whary, et al. (2005), Infect Immun 73(3): 1664-70. Abstract: Persistent Helicobacter felis infection in (C57BL/6 x 129SvEv)F1 mice induces chronic gastritis. Expression of inducible nitric oxide synthase (iNOS) is upregulated in response to Helicobacter infection. In this study, 20 10-week-old iNOS-/- mice and 20 wild-type (C57BL/6 x 129SvEv)F1 mice were infected with H. felis by oral gavage and were assessed histologically and serologically at 32 weeks postinfection. Equal numbers of uninfected controls were sham inoculated. The mice were scored for severity of gastric inflammation, hyperplasia, glandular atrophy, and mucous metaplasia in the corpus and for the level of helicobacter colonization. The immunoglobulin G1 (IgG1), IgG2a, and IgG2c antibody responses to H. felis were determined. As a secondary measure, serum cholesterol levels were assessed. iNOS-/- mice have a propensity for increased serum cholesterol, and although controversial, several human epidemiologic studies have demonstrated an association between Helicobacter infection and several risk factors for cardiovascular disease, including elevated serum cholesterol. Nevertheless, no differences in serum cholesterol levels were observed between the H. felis-infected and -uninfected iNOS-/- mice in this study. The uninfected animals had minimal to no gastric pathology. The gastric pathology scores for the infected animals were reduced significantly in the iNOS-deficient mice relative to those for the wild-type mice (all P <0.01). Helicobacter-infected iNOS-/- mice had chronic lymphoid infiltration and negligible to mild glandular atrophy and mucous metaplasia in the fundic mucosa, while H. felis-infected wild-type mice had severe atrophic and metaplastic mucosal changes. The atrophic gastritis in the infected wild-type mice, particularly the female mice, was also accompanied by greater granulocytic infiltration, antral hyperplasia, and diminished antral colonization, unlike that in the infected iNOS-/- mice. iNOS-/- mice developed significantly lower Th1-associated IgG2c antibody responses to H. felis (P <0.0003); the Th2-associated IgG1 responses were similar (P=0.09), suggesting a greater effect of the iNOS defect on Th1 responses. H. felis colonization was significantly greater in the iNOS-deficient mice. These findings are indicative of an impaired Th1 component of the H. felis-induced inflammatory response when the influence of iNOS is removed. |