Bioluminescence measurement of primary bile acids using immobilized 7 alpha-hydroxysteroid dehydrogenase: application to serum bile acids.

Abstract:

:A simple, rapid, and sensitive bioluminescence method for measuring primary bile acids has been developed and validated. The method is based on enzymatic dehydrogenation of bile acids using a bacterial 7 alpha-hydroxysteroid dehydrogenase that is co-immobilized on Sepharose 4B beads with NADH:FMN oxidoreductase and a bacterial luciferase. The assay is specific for 7 alpha-hydroxy bile acids and has a detection limit of 0.5 pmol/tube, with a linear range of 0.5-50 pmol/tube. The assay shows good precision (6-8% intra-assay; 8-10% inter-assay). The values obtained with the bioluminescence assay showed good agreement with those obtained by gas-liquid chromatography, radioimmunoassay, or endpoint enzymatic assays. When applied to the measurement of serum bile acids, there was no interference from serum albumin, and the effect of other dehydrogenase activity in serum could be eliminated by heating the sample prior to assay. Since the method is rapid (1 minute), extremely sensitive (requires only 10 microliters of serum), and specific, it appears to be the best method currently available for the measurement of serum primary bile acids.

journal_name

J Lipid Res

authors

Roda A,Kricka LJ,DeLuca M,Hofmann AF

subject

Has Abstract

pub_date

1982-12-01 00:00:00

pages

1354-61

issue

9

eissn

0022-2275

issn

1539-7262

journal_volume

23

pub_type

杂志文章
  • Metabolism of oxysterols derived from nonenzymatic oxidation of 7-dehydrocholesterol in cells.

    abstract::Recent studies suggest that 7-dehydrocholesterol (7-DHC)-derived oxysterols play important roles in the pathophysiology of Smith-Lemli-Opitz syndrome (SLOS), a metabolic disorder that is caused by defective 3β-hydroxysterol-Δ(7)-reductase (DHCR7). Although 14 oxysterols have been identified as the primary products of ...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:10.1194/jlr.M035733

    authors: Xu L,Korade Z,Rosado DA Jr,Mirnics K,Porter NA

    更新日期:2013-04-01 00:00:00

  • Separation of gluco- and galactocerebrosides by means of borate thin-layer chromatography.

    abstract::Gluco- and galactocerebrosides can be separated by thin-layer chromatography on Silica Gel G prepared with sodium borate solution instead of water. The most successful developing system was chloroform-methanol-water-15 M NH(4)OH 280:70:6:1. ...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: Kean EL

    更新日期:1966-05-01 00:00:00

  • Dephosphorylation and benzoylation reactions occurring in the benzoolysis of diacyl-, alkylacyl-, alk-1-enylacyl-, and acyl-lyso-glycerophosphocholines.

    abstract::Series of benzoolysis experiments with 1,2-dipalmitoyl glycerophosphocholine (GPC), 1-palmitoyl-2-lyso GPC, and 1-O-hexadecenyl-2-oleoyl GPC, as well as a few experiments with 1-oleoyl-2-acetyl GPC and 1-O-hexadecyl-2-acetyl GPC are reported. The results are used to present a consistent picture of the various dephosph...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: Gelsema WJ,van den Bosch H

    更新日期:1997-08-01 00:00:00

  • A facile synthesis of [14C] epicholesterol.

    abstract::A facile procedure is described for the preparation of [14C]epicholesterol from [14C]cholesterol. Cholesterol is first converted to cholesteryl mesylate, which is treated with cesium acetate and 18-crown-6 in refluxing toluene to give epicholesteryl acetate. The latter is hydrolyzed, without isolation, with potassium ...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: Yan JS,Bittman R

    更新日期:1990-01-01 00:00:00

  • Influence of dietary trans-fatty acids on swine lipoprotein composition and structure.

    abstract::Four groups of 20 weanling swine each were fed either (a) basal diet, (b) basal plus hydrogenated fat (13% trans), (c) basal plus hydrogenated fat (13% trans) and 0.4% cholesterol, or (d) basal plus beef tallow (all cis). After six months of feeding, the animals were killed and the blood and aortas were removed. Very ...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: Jackson RL,Morrisett JD,Pownall HJ,Gotto AM Jr,Kamio A,Imai H,Tracy R,Kummerow FA

    更新日期:1977-03-01 00:00:00

  • Bile acid synthesis in the Smith-Lemli-Opitz syndrome: effects of dehydrocholesterols on cholesterol 7alpha-hydroxylase and 27-hydroxylase activities in rat liver.

    abstract::The Smith-Lemli-Opitz syndrome (SLOS) is a congenital birth defect syndrome caused by a deficiency of 3beta-hydroxysterol Delta(7)-reductase, the final enzyme in the cholesterol biosynthetic pathway. The patients have reduced plasma and tissue cholesterol concentrations with the accumulation of 7-dehydrocholesterol an...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: Honda A,Salen G,Shefer S,Batta AK,Honda M,Xu G,Tint GS,Matsuzaki Y,Shoda J,Tanaka N

    更新日期:1999-08-01 00:00:00

  • Apolipoprotein B(Arg3500----Gln) allele specific polymerase chain reaction: large-scale screening of pooled blood samples.

    abstract::A two-step polymerase chain reaction (PCR) method for the rapid detection of the apolipoprotein B(Arg3500----Gln) mutation in a mixture of pooled blood samples is described. In the first step PCR, a short gene fragment surrounding codon 3500 is amplified. Subsequently the reaction product is subjected to a second ampl...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: Ruzicka V,März W,Russ A,Gross W

    更新日期:1992-10-01 00:00:00

  • Recognition and receptor-mediated endocytosis of the lysosomal acid lipase secreted by cultured human fibroblasts.

    abstract::We have studied the recognition and uptake of acid lipase by human fibroblasts in order to determine requirements for localization and function of the enzyme in lysosomes. Our approach was based on evidence that a number of acid hydrolases involved in mucopolysaccharide metabolism are secreted from cultured fibroblast...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: Sando GN,Henke VL

    更新日期:1982-01-01 00:00:00

  • Regulation of hepatic cholesterol ester hydrolase and acyl-coenzyme A:cholesterol acyltransferase in the rat.

    abstract::Cholesterol exists within the hepatocyte as free cholesterol and cholesteryl ester. The proportion of intrahepatic cholesterol in the free or ester forms is governed in part by the rate of cholesteryl ester formation by acyl-coenzyme A:cholesterol acyltransferase (ACAT) and cholesteryl ester hydrolysis by neutral chol...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: Stone BG,Evans CD,Fadden RJ,Schreiber D

    更新日期:1989-11-01 00:00:00

  • Identification of enzymes involved in oxidation of phenylbutyrate.

    abstract::In recent years the short-chain fatty acid, 4-phenylbutyrate (PB), has emerged as a promising drug for various clinical conditions. In fact, PB has been Food and Drug Administration-approved for urea cycle disorders since 1996. PB is more potent and less toxic than its metabolite, phenylacetate (PA), and is not just a...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:10.1194/jlr.M075317

    authors: Palir N,Ruiter JPN,Wanders RJA,Houtkooper RH

    更新日期:2017-05-01 00:00:00

  • PIP4K2A regulates intracellular cholesterol transport through modulating PI(4,5)P2 homeostasis.

    abstract::The transport of LDL-derived cholesterol from lysosomes to peroxisomes is facilitated by membrane contacts formed between the lysosomal protein synaptotagmin VII and the peroxisomal lipid phosphatidylinositol 4, 5-bisphosphate [PI(4,5)P2]. Here, we used RNA interference to search for regulators of PI(4,5)P2 and to stu...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:10.1194/jlr.M082149

    authors: Hu A,Zhao XT,Tu H,Xiao T,Fu T,Wang Y,Liu Y,Shi XJ,Luo J,Song BL

    更新日期:2018-03-01 00:00:00

  • Chylomicron apoprotein localization within rat intestinal epithelium: studies of normal and impaired lipid absorption.

    abstract::Monospecific antisera were produced to two chylomicron apoproteins (apoB, apoA-I) and utilized for indirect immunofluorescent localization of these apoproteins within rat intestinal epithelium during normal and impaired lipid absorption. Isolated intestinal epithelial cells prepared after different periods of lipid ab...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: Glickman RM,Kilgore A,Khorana J

    更新日期:1978-02-01 00:00:00

  • CDP-diacylglycerol synthases regulate the growth of lipid droplets and adipocyte development.

    abstract::The expansion of lipid droplets (LDs) and the differentiation of preadipocytes are two important aspects of mammalian lipid storage. In this study, we examined the role of CDP-diacylglycerol (DAG) synthases (CDSs), encoded by CDS1 and CDS2 genes in mammals, in lipid storage. CDS enzymes catalyze the formation of CDP-D...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:10.1194/jlr.M060574

    authors: Qi Y,Kapterian TS,Du X,Ma Q,Fei W,Zhang Y,Huang X,Dawes IW,Yang H

    更新日期:2016-05-01 00:00:00

  • Glucose stimulates cholesterol 7alpha-hydroxylase gene transcription in human hepatocytes.

    abstract::Bile acids play important roles in the regulation of lipid, glucose, and energy homeostasis. Recent studies suggest that glucose regulates gene transcription in the liver. The aim of this study was to investigate the potential role of glucose in regulation of bile acid synthesis in human hepatocytes. High glucose stim...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:10.1194/jlr.M002782

    authors: Li T,Chanda D,Zhang Y,Choi HS,Chiang JY

    更新日期:2010-04-01 00:00:00

  • Effects of lovastatin and chenodiol on bile acid synthesis, bile lipid composition, and biliary lipid secretion in healthy human subjects.

    abstract::To assess the relationship between cholesterol synthesis and feedback inhibition of bile acid synthesis, we studied seven normal human subjects taking three different doses of chenodiol, 0, 5, and 15 mg/kg per day: once while taking no lovastatin and again while taking lovastatin 80 mg/day. Lovastatin and both doses o...

    journal_title:Journal of lipid research

    pub_type: 临床试验,杂志文章,随机对照试验

    doi:

    authors: Hanson DS,Duane WC

    更新日期:1994-08-01 00:00:00

  • Lipid-induced NOX2 activation inhibits autophagic flux by impairing lysosomal enzyme activity.

    abstract::Autophagy is a catabolic process involved in maintaining energy and organelle homeostasis. The relationship between obesity and the regulation of autophagy is cell type specific. Despite adverse consequences of obesity on cardiac structure and function, the contribution of altered cardiac autophagy in response to fatt...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:10.1194/jlr.M055152

    authors: Jaishy B,Zhang Q,Chung HS,Riehle C,Soto J,Jenkins S,Abel P,Cowart LA,Van Eyk JE,Abel ED

    更新日期:2015-03-01 00:00:00

  • Combined (n-3 and n-6) essential fatty deficiency is a potent modulator of plasma lipids, lipoprotein composition, and lipolytic enzymes.

    abstract::Essential fatty acids (EFA) are important structural and functional components of cell membranes. Their deficiency has been associated with several clinical and biochemical abnormalities. In the present study, the lipid profile as well as the concentration, composition, and metabolism of lipoproteins were examined in ...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: Levy E,Thibault L,Garofalo C,Messier M,Lepage G,Ronco N,Roy CC

    更新日期:1990-11-01 00:00:00

  • Lipolysis of very low density lipoproteins by heparan sulfate proteoglycan-bound lipoprotein lipase.

    abstract::An in vitro assay to study lipolysis of very low density lipoproteins (VLDL) by heparan sulfate proteoglycan (HSPG-bound lipoprotein lipase (LPL) was developed. Optimal conditions for VLDL lipolysis by HSPG-bound LPL were obtained by incubating plastic wells with 0.5 microg HSPG and 1.5 microg LPL, subsequently. Contr...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: de Man FH,de Beer F,van der Laarse A,Smelt AH,Havekes LM

    更新日期:1997-12-01 00:00:00

  • A fluorescent assay to quantitatively measure in vitro acyl CoA:diacylglycerol acyltransferase activity.

    abstract::Triacylglycerols (TG) are the major storage form of energy in eukaryotic organisms and are synthesized primarily by acyl CoA:1,2-diacylglycerol acyltransferase (DGAT) enzymes. In vitro DGAT activity has previously been quantified by measuring the incorporation of either radiolabeled fatty acyl CoA or diacylglycerol (D...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:10.1194/jlr.D016626

    authors: McFie PJ,Stone SJ

    更新日期:2011-09-01 00:00:00

  • Adipose tissue cellularity and histochemistry in fetal swine as affected by genetic selection for high or low backfat.

    abstract::Adipose tissue development was studied histochemically and histologically in fetuses from lean and obese sows. At 110 days of gestation, fetuses were removed from Ossabaw (obese-feral) sows and from sows selected for high backfat (obese-domestic) and for low backfat (lean) thickness. Body weights were similar for lean...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: Hausman GJ,Campion DR,Thomas GB

    更新日期:1983-03-01 00:00:00

  • Simulation of triacylglycerol ion profiles: bioinformatics for interpretation of triacylglycerol biosynthesis.

    abstract::Although the synthesis pathways of intracellular triacylglycerol (TAG) species have been well elucidated, assessment of the contribution of an individual pathway to TAG pools in different mammalian organs, particularly under pathophysiological conditions, is difficult, although not impossible. Herein, we developed and...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:10.1194/jlr.M033837

    authors: Han RH,Wang M,Fang X,Han X

    更新日期:2013-04-01 00:00:00

  • A single high-cholesterol, high-fat meal preferentially increases low molecular weight apolipoprotein B concentration in rat plasma.

    abstract::Rats were fed either rat chow (control), chow + 20% olive oil (olive oil), or chow + 20% olive oil + 2% cholesterol (olive oil/cholesterol) as a single meal to study the short-term effects of fat and the above combination of fat/cholesterol-containing diets on plasma apoB concentration and its influence on the distrib...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: DeLamatre JG,Krause BR,Wong L,Hoffmeier CA,Roheim PS

    更新日期:1985-08-01 00:00:00

  • Turnover of endogenous ceramide in cultured normal and Farber fibroblasts.

    abstract::De novo synthesis and turnover of endogenous ceramide in cultured skin fibroblasts from patients affected with Farber lipogranulomatosis were studied by biosynthetical labeling of cellular sphingolipids with [14C]serine. The cellular uptake of [14C]serine and incorporation into de novo synthesized ceramide was similar...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: van Echten-Deckert G,Klein A,Linke T,Heinemann T,Weisgerber J,Sandhoff K

    更新日期:1997-12-01 00:00:00

  • LY295427, a novel hypocholesterolemic agent, enhances [3H]25-hydroxycholesterol binding to liver cytosolic proteins.

    abstract::LY295427, (3 alpha,4 alpha,5 alpha)-4-(2-propenylcholestan-3-ol), acts through an unknown mechanism to derepress the transcription of the low density lipoprotein (LDL) receptor in the presence of 25-hydroxycholesterol (25-OH chol). Preincubation with LY295427 in Chinese hamster ovary (CHO) cells increased uptake of 25...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: Bowling N,Matter WF,Gadski RA,McClure DB,Schreyer T,Dawson PA,Vlahos CJ

    更新日期:1996-12-01 00:00:00

  • Enzymatic assay for cholesterol ester hydrolase activity.

    abstract::A rapid and accurate method is described for the assay of cholesterol ester hydrolase (CEH) activity. Aliquots of the enzyme-substrate incubation mixture are extracted into isopropanol. The free cholesterol concentration in each extract is determined enzymatically using a single aqueous reagent containing cholesterol ...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: Gallo LL,Atasoy R,Vahouny GV,Treadwell CR

    更新日期:1978-09-01 00:00:00

  • Hormone-sensitive lipase expression and activity in relation to lipolysis in human fat cells.

    abstract::Hormone-sensitive lipase (HSL) catalyzes the rate-limiting step in adipocyte lipolysis. The activity of HSL is thought to be primarily regulated by reversible phosphorylation. However, the regulation of HSL activity by pre-translational mechanisms has been poorly studied. The present studies were undertaken to explore...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: Large V,Arner P,Reynisdottir S,Grober J,Van Harmelen V,Holm C,Langin D

    更新日期:1998-08-01 00:00:00

  • Plasma levels of lecithin:cholesterol acyltransferase and risk of future coronary artery disease in apparently healthy men and women: a prospective case-control analysis nested in the EPIC-Norfolk population study.

    abstract::LCAT plays a key role in the maturation of HDL, as evidenced by low HDL-cholesterol levels in carriers of deleterious mutations in LCAT. However, the role of LCAT in atherosclerosis is unclear. We set out to study this in a prospective study. Plasma LCAT levels, which strongly correlate with LCAT activity, were measur...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:10.1194/P900038-JLR200

    authors: Holleboom AG,Kuivenhoven JA,Vergeer M,Hovingh GK,van Miert JN,Wareham NJ,Kastelein JJ,Khaw KT,Boekholdt SM

    更新日期:2010-02-01 00:00:00

  • Lipid accumulation in isolated perfused rat hearts has no apparent effect on mechanical function or energy metabolism as measured by 31P NMR.

    abstract::Male Sprague-Dawley rats were fed diets that contained 20% by weight soybean oil or rapeseed oil (21% and 43% erucic acid) for 7 days. The rapeseed oil diets increased the cardiac triacylglycerol content 5-fold and 25-fold, respectively, above control values. Hearts were removed from the animals and perfused with modi...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:

    authors: Stewart LC,Kramer JK,Sauer FD,Clarke K,Wolynetz MS

    更新日期:1993-09-01 00:00:00

  • Hypercholesterolemia and changes in lipid and bile acid metabolism in male and female cyp7A1-deficient mice.

    abstract::Cholesterol 7alpha-hydroxylase, a rate-limiting enzyme for bile acid synthesis, has been implicated in genetic susceptibility to atherosclerosis. The gene, CYP7A1, encoding a protein with this activity, is expressed normally only in hepatocytes and is highly regulated. Our cyp7A1 gene knockout mouse colony, as young a...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:10.1194/jlr.M200489-JLR200

    authors: Erickson SK,Lear SR,Deane S,Dubrac S,Huling SL,Nguyen L,Bollineni JS,Shefer S,Hyogo H,Cohen DE,Shneider B,Sehayek E,Ananthanarayanan M,Balasubramaniyan N,Suchy FJ,Batta AK,Salen G

    更新日期:2003-05-01 00:00:00

  • PPARalpha activation increases triglyceride mass and adipose differentiation-related protein in hepatocytes.

    abstract::Adipose differentiation-related protein (ADRP) is a lipid droplet-associated protein that is expressed in various tissues. In mice treated with the peroxisome proliferator-activated receptor alpha (PPARalpha) agonist Wy14,643 (Wy), hepatic mRNA and protein levels of ADRP as well as hepatic triglyceride content increas...

    journal_title:Journal of lipid research

    pub_type: 杂志文章

    doi:10.1194/jlr.M500203-JLR200

    authors: Edvardsson U,Ljungberg A,Lindén D,William-Olsson L,Peilot-Sjögren H,Ahnmark A,Oscarsson J

    更新日期:2006-02-01 00:00:00