Flow-cytometric determination of high-density-lipoprotein binding sites on human leukocytes.

Abstract:

:In this method, leukocytes were isolated from 6 mL of EDTA-blood by density-gradient centrifugation and subsequently incubated with rhodamine isothiocyanate (RITC)-conjugated high-density lipoproteins (HDL). The receptor-bound conjugate particles were determined by fluorescent flow cytometry and compared with 125I-labeled HDL binding data for the same cells. Human granulocytes express the highest number of HDL binding sites (9.4 X 10(4)/cell), followed by monocytes (7.3 X 10(4)/cell) and lymphocytes (4.0 X 10(4)/cell). Compared with conventional analysis of binding of 125I-labeled HDL in tissue-culture dishes, the present determination revealed significantly lower values for nonspecific binding. In competition studies, the conjugate competes for the same binding sites as 125I-labeled HDL. With the use of tetranitromethane-treated HDL3, which fails to compete for the HDL receptor sites while nonspecific binding is not affected, we could clearly distinguish between 37 degrees C surface binding and specific 37 degrees C uptake of RITC-HDL3, confirming that the HDL receptor leads bound HDL particles into an intracellular pathway rather than acting as a "docking" type of receptor. Patients with familial dysbetalipoproteinemia showed a significantly higher number of HDL binding sites in the granulocyte population but normal in lymphocytes and monocytes, indicating increased uptake of cholesterol-containing lipoproteins. In patients with familial hypercholesterolemia, HDL binding was increased in all three cell types, indicating increased cholesterol uptake and increased cholesterol synthesis. The present method allows rapid determination of HDL binding sites in leukocytes from patients with various forms of hyper- and dyslipoproteinemias.

journal_name

Clin Chem

journal_title

Clinical chemistry

authors

Schmitz G,Wulf G,Brüning T,Assmann G

subject

Has Abstract

pub_date

1987-12-01 00:00:00

pages

2195-203

issue

12

eissn

0009-9147

issn

1530-8561

journal_volume

33

pub_type

杂志文章
  • Immunoradiometric assay for alpha gamma- and gamma gamma-enolase (neuron-specific enolase), with use of monoclonal antibodies and magnetizable polymer particles.

    abstract::Monoclonal antibodies were raised against neuron-specific enolase, gamma gamma-enolase, and used in an immunoradiometric assay (IRMA), with mono-disperse magnetizable particles as the solid phase. The assay's sensitivity was 0.4 microgram/L and the interassay coefficient of variation was less than 5% in the working ra...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Paus E,Nustad K

    更新日期:1989-10-01 00:00:00

  • Rapid liquid chromatography-tandem mass spectrometry method for routine analysis of cyclosporin A over an extended concentration range.

    abstract:BACKGROUND:Cyclosporin A (CsA) is commonly measured by immunoassay techniques that have a limited analytical range. The consequence of this is that low CsA concentrations that may be clinically significant are difficult to measure and that high concentrations require sample dilution, which introduces error and increase...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Keevil BG,Tierney DP,Cooper DP,Morris MR

    更新日期:2002-01-01 00:00:00

  • Comparison between immunoturbidimetry, size-exclusion chromatography, and LC-MS to quantify urinary albumin.

    abstract:BACKGROUND:The accurate and precise measurement of urinary albumin is critical, since even minor increases are diagnostically sensitive indicators of renal disease, cardiovascular events, and risk for death. To gain insights into potential measurement biases, we systematically compared urine albumin measurements perfor...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2008.107508

    authors: Shaikh A,Seegmiller JC,Borland TM,Burns BE,Ladwig PM,Singh RJ,Kumar R,Larson TS,Lieske JC

    更新日期:2008-09-01 00:00:00

  • Beyond diagnostic accuracy: the clinical utility of diagnostic tests.

    abstract::Like any other medical technology or intervention, diagnostic tests should be thoroughly evaluated before their introduction into daily practice. Increasingly, decision makers, physicians, and other users of diagnostic tests request more than simple measures of a test's analytical or technical performance and diagnost...

    journal_title:Clinical chemistry

    pub_type: 杂志文章,评审

    doi:10.1373/clinchem.2012.182576

    authors: Bossuyt PM,Reitsma JB,Linnet K,Moons KG

    更新日期:2012-12-01 00:00:00

  • A parallel revolution: payment for new in vitro diagnostic technologies.

    abstract::To secure a successful product in today's era of cost containment, manufacturers and developers of new in vitro diagnostic technologies must consider payment issues--coverage and reimbursement by healthcare insurers--as critical market forces that must be addressed in the business plan. Similar to the regulatory hurdl...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Ivor LP

    更新日期:1996-09-01 00:00:00

  • Coagulation and bleeding disorders: review and update.

    abstract::Hemostasis is initiated by injury to the vascular wall, leading to the deposition of platelets adhering to components of the subendothelium. Platelet adhesion requires the presence of von Willebrand factor and platelet receptors (IIb/IIIa and Ib/IX). Additional platelets are recruited to the site of injury by release ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章,评审

    doi:

    authors: Triplett DA

    更新日期:2000-08-01 00:00:00

  • Surface modification with protein A for uniform binding of monoclonal antibodies.

    abstract::We describe optimal conditions for immobilization of two monoclonal antibodies to progesterone for solid-phase assays. Polystyrene surfaces are refined with Protein A to achieve uniform, reproducible, stable, and sterically accessible immobilization of immunoglobulins (IgG). To this end, we optimized the amount of imm...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Schramm W,Yang T,Midgley AR

    更新日期:1987-08-01 00:00:00

  • Cardiac troponin-I is not expressed in fetal and healthy or diseased adult human skeletal muscle tissue.

    abstract::Cardiac troponin-I (cTnI) is not found in sera of patients with skeletal muscle disease in the absence of myocardial injury. It is not known, however, whether trace amounts of cTnI are expressed in regenerating human skeletal muscle, as has been observed with creatine kinase MB. Using immunohistochemical and biochemic...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Bodor GS,Porterfield D,Voss EM,Smith S,Apple FS

    更新日期:1995-12-01 00:00:00

  • Quantitative PCR analysis of DNA, RNAs, and proteins in the same single cell.

    abstract:BACKGROUND:The single cell represents the basic unit of all organisms. Most investigations have been performed on large cell populations, but understanding cell dynamics and heterogeneity requires single-cell analysis. Current methods for single-cell analysis generally can detect only one class of analytes. METHODS:Re...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2012.191445

    authors: Ståhlberg A,Thomsen C,Ruff D,Åman P

    更新日期:2012-12-01 00:00:00

  • Meconium nicotine and metabolites by liquid chromatography-tandem mass spectrometry: differentiation of passive and nonexposure and correlation with neonatal outcome measures.

    abstract:BACKGROUND:Meconium analysis is a diagnostically sensitive and objective alternative to maternal self-report for detecting prenatal tobacco exposure. Nicotine and metabolite disposition in meconium is poorly characterized, and correlation of analytes' concentrations with neonatal outcomes is unexplored. Our objectives ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2008.109173

    authors: Gray TR,Magri R,Shakleya DM,Huestis MA

    更新日期:2008-12-01 00:00:00

  • "Delfia" and "Amerlite": two sensitive nonisotopic immunoassay systems for assay of thyrotropin compared.

    abstract::We assessed the LKB "Delfia" (time-resolved dissociation-enhanced lanthanide fluoroimmunoassay) and the Amersham "Amerlite" (enhanced luminescent immunometry) assays of thyrotropin in serum. Both assays are sensitive (respective detection limits: 0.02 and 0.04 milli-int. unit/L) and have very good within- and between-...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Parnham AJ,Tarbit IF

    更新日期:1987-08-01 00:00:00

  • Multilayer film analysis for glucose in 1-microL samples of plasma.

    abstract::With the ultramicroanalytical system described here we can measure glucose in 1 microL of plasma or serum. The sample is placed on a dry, multilayer film element (Eastman Kodak), where a colored spot about 3.5 mm in diameter develops. The reflectance of these spots is measured with a reflectance digital matrix photome...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Neeley WE,Zettner A

    更新日期:1983-12-01 00:00:00

  • Quantification of urinary porphyrins by liquid chromatography after oxidation of porphyrinogens.

    abstract::A quantitative "high-performance" liquid-chromatographic method is described for determining porphyrins in human urine. Porphyrinogens in urine are first converted to the corresponding porphyrins by oxidation with iodine. Uroporphyrin, hepatacarboxylic acid porphyrin, hexacarboxylic acid porphyrin, pentacarboxylic aci...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Abe K,Konaka R

    更新日期:1989-08-01 00:00:00

  • Micro-scale method for theophylline in body fluids by reversed-phase, high-pressure liquid chromatography.

    abstract::We describe a micro-scale method for determining serum theophylline. The chromatography system includes a muBondapack C18 column and acetonitrile, 70 ml/liter of sodium acetate buffer (10 mmol/liter, ph 4.0) as the mobile phase. Test serum or plasma, 30 mul, is mixed with an equal quantity of a solution containing the...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Orcutt JJ,Kozak PP Jr,Gillman SA,Cummins LH

    更新日期:1977-03-01 00:00:00

  • Determination of cyclosporine concentrations with monoclonal antibodies.

    abstract::We measured cyclosporine in whole blood from normal volunteers administered single oral doses of the drug and from two renal-transplant patients on immunosuppressive maintenance therapy, by liquid chromatography (I) and by radioimmunoassay with use of nonspecific polyclonal (II), specific monoclonal (III), and nonspec...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Schran HF,Rosano TG,Hassell AE,Pell MA

    更新日期:1987-12-01 00:00:00

  • Analysis for lead in undiluted whole blood by tantalum ribbon atomic absorption spectrophotometry.

    abstract::We describe a modified tantalum ribbon atomic absorption procedure for determining lead in undiluted whole blood. An instrumentation Laboratory (I.L.) Model 151 atomic absorption spectrophotometer equipped with an I.L. Model 355 Flameless Sampler was used. The Flameless Sampler was slightly modified to include three-c...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Therrell BL Jr,Drosche JM,Dziuk TW

    更新日期:1978-07-01 00:00:00

  • Calorimetry of cholinesterase activity in serum.

    abstract::Cholinesterase activity in human serum was measured calorimetrically and compared to values obtained with the spectrophotometric method of Rappaport et al. [Clin. Chim. Acta 4, 227 (1959)]. The response of the calorimeter upon mixing serum with acetylcholine corresponded to the cholinesterase activity of the serum, th...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: O'Farrell HK,Chattopadhyay SK,Brown HD

    更新日期:1977-10-01 00:00:00

  • Diagnosis of dopa-responsive dystonia and other tetrahydrobiopterin disorders by the study of biopterin metabolism in fibroblasts.

    abstract:BACKGROUND:Dopa-responsive dystonia (DRD) and tetrahydrobiopterin (BH4) defects are inherited disorders characterized by monoamine neurotransmitter deficiency with decreased activity of one of the BH4-metabolizing enzymes. The aim of the study was to determine the utility of cultured skin fibroblasts for the diagnosis ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Bonafé L,Thöny B,Leimbacher W,Kierat L,Blau N

    更新日期:2001-03-01 00:00:00

  • Separation of bound and free ligand by ethacridine (Rivanol) in thyroxin radioimmunoassay.

    abstract::In this simple and low-cost radioimmunoassay (RIA) procedure for thyroxin (T4), ethacridine (6,9-diamino-2-ethoxyacridine lactate; Rivanol) is used to separate antibody-bound ligand and free ligand. This acridine dye precipitates free T4 not bound to antibody. At pH 8.6, it precipitates more than 90% of 125I-labeled T...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Bhupal V,Mani RS

    更新日期:1983-11-01 00:00:00

  • Differential gene expression of Eph receptors and ephrins in benign human tissues and cancers.

    abstract:BACKGROUND:Eph receptors and their ligands, the ephrins, represent a large class of cell-cell communication molecules with well-defined developmental functions. Their role in healthy adult tissues and in human disease is still largely unknown, although diverse roles in carcinogenesis have been postulated. METHODS:We e...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2003.026849

    authors: Hafner C,Schmitz G,Meyer S,Bataille F,Hau P,Langmann T,Dietmaier W,Landthaler M,Vogt T

    更新日期:2004-03-01 00:00:00

  • Population-based study of repeat laboratory testing.

    abstract:BACKGROUND:Test repetition could be a readily modifiable component of laboratory utilization. Laboratory test repetition has not been rigorously studied at a population-based level. Our objective was to determine the prevalence of, and charges associated with, repetition of eight common laboratory tests. METHODS:We pe...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2003.021220

    authors: van Walraven C,Raymond M

    更新日期:2003-12-01 00:00:00

  • Flow-injection analysis for malondialdehyde in plasma with the thiobarbituric acid reaction.

    abstract::A simple, precise, and rapid method to measure plasma malondialdehyde (MDA) was developed by use of solvent extraction--flow-injection analysis. The reagent solution, containing thiobarbituric acid (TBA), 5 g/L in 100 mL/L phosphoric acid, and extraction solvent (methylisobutyl ketone, MIBK) were propelled with a doub...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Ikatsu H,Nakajima T,Murayama N,Korenaga T

    更新日期:1992-10-01 00:00:00

  • Denaturing HPLC profiling of the ABCA4 gene for reliable detection of allelic variations.

    abstract:BACKGROUND:Mutations in the retina-specific ABC transporter (ABCA4) gene have been associated with several forms of macular degenerations. Because the high complexity of the molecular genotype makes scanning of the ABCA4 gene cumbersome, we describe here the first use of denaturing HPLC (DHPLC) to screen for ABCA4 muta...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2004.033241

    authors: Stenirri S,Fermo I,Battistella S,Galbiati S,Soriani N,Paroni R,Manitto MP,Martina E,Brancato R,Allikmets R,Ferrari M,Cremonesi L

    更新日期:2004-08-01 00:00:00

  • Inhibition of stimulated interleukin-2 production in whole blood: a practical measure of cyclosporine effect.

    abstract:BACKGROUND:Prediction of cyclosporine (CSA) efficacy and toxicity in individual patients is difficult. There is no practical, biologically relevant, pharmacodynamic measure of CSA effect. A major effect of CSA is to decrease interleukin-2 (IL-2) production; however, measurement of this effect in isolated lymphocytes as...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Stein CM,Murray JJ,Wood AJ

    更新日期:1999-09-01 00:00:00

  • Estimating fat in feces by near-infrared reflectance spectroscopy.

    abstract::We describe use of near-infrared reflectance spectroscopy (NIRRS) to estimate the amount of fat in feces, for diagnosis of steatorrhea. After sample homogenization, the spectrum of the fecal homogenate is scanned over the near-infrared region. Assay of 94 samples of feces having a known concentration of fat showed the...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Koumantakis G,Radcliff FJ

    更新日期:1987-04-01 00:00:00

  • Radioimmunoassay of creatine kinase isoenzymes in human serum: isoenzyme MM.

    abstract::Measurement of the mass concentration of serum enzymes by radioimmunoassay provides direct quantitation of specific isoenzymes and may be less subject to some of the limitations of traditional assay procedures for enzymes. We describe the development of a sensitive and specific radioimmunoassay for the muscle isoenzym...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Van Steirteghem AC,Zweig MH,Schechter AN

    更新日期:1978-03-01 00:00:00

  • Analysis for diazepam and nordiazepam by electron-capture gas chromatography and by liquid chromatography.

    abstract::We describe the use of electron-capture gas chromatography or reversed-phase "high-performance" liquid chromatography for concurrent analysis for diazepam and nordiazepam in serum. In the gas-chromatographic analysis our use of a new chemically deactivated stationary liquid phase, SP 2250-DB (Supelco, Inc.), resulted ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Wallace JE,Schwertner HA,Shimek EL Jr

    更新日期:1979-07-01 00:00:00

  • Theoretical aspects of one-point calibration: causes and effects of some potential errors, and their dependence on concentration.

    abstract::The potential errors of the one-point calibration technique are described mathematically. I examine the theoretical causes and effects of analytical errors in absorbance and of errors in determining calibration concentration, and discuss techniques for minimizing their impact. The dependence of these errors on the cal...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Kemp GJ

    更新日期:1984-07-01 00:00:00

  • Determination of tamoxifen and four metabolites in serum by low-dispersion liquid chromatography.

    abstract::In this assay of tamoxifen and four metabolites in human serum, the serum samples are deproteinized with an equal volume of acetonitrile, then injected into a small (0.21 X 2 cm) precolumn packed with 5-micron-diameter octadecylsilane (ODS) particles. The samples are concentrated on-column by equilibrating the column ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Lien EA,Ueland PM,Solheim E,Kvinnsland S

    更新日期:1987-09-01 00:00:00

  • Plasma protein abnormalities in nephrotic syndrome: effect on plasma colloid osmotic pressure and viscosity.

    abstract::The concentrations of 25 plasma proteins were measured in 22 patients with membranous nephropathy. For some large proteins, the plasma concentrations were increased; there were also large proteins with low plasma concentrations, but small or medium-sized proteins showed uniformly lower plasma concentration than the co...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Joven J,Clivillé X,Camps J,Espinel E,Simó J,Vilella E,Oliver A

    更新日期:1997-07-01 00:00:00