bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analysis of human leukemic cells by use of high-resolution two-dimensional electrophoresis. I: results of a pilot study.
Authors: Anderson N L , Wiltsie J C , Li C Y , Willard-Gallo K E , Tracy R P , Young D S , Powers M T , Anderson N G 
Published: January 2020

Abstract
We analyzed mononuclear leukocytes from patients with various human leukemias by high-resolution two-dimensional electrophoresis. Tumor cells of the granulocytic, monocytic, and lymphoid lineages [obtained from chronic granulocytic leukemia in blast transformation, acute monocytic leukemia, and chronic lymphocytic leukemia (CLL), respectively] can be easily recognized by using a series of cell-type marker proteins identified by comparison of fractionated normal cell populations. B and T cell types of CLL could be distinguished, the results correlating well with those obtained by use of monoclonal-antibody staining methods. In two cases representing almost pure B-cells (classical CLL; 0% T, 85% B) and T-cells (cutaneous T-cell leukemia; 77% T, 0% B), 27 of 29 marker proteins showed quantitative B/T differences comparable to those observed in comparisons of normal B-and T-lymphocytes prepared by cell sorting. These results indicate that cells from relatively well-differentiated leukemias show complex patterns of gene expression very similar to those of the corresponding normal cells and strongly support the use of large marker panels in cell-type determination. Less-well-differentiated acute leukemias [such as acute undifferentiated and acute granulocytic (FAB:M1)] appear to yield protein patterns corresponding less closely to recognizable mature cell types, and may show expression of novel proteins related to the state of differentiation.
Serum lipoprotein measurement--liquid chromatography and sequential floatation (ultracentrifugation) compared.
Authors: Okazaki M , Itakura H , Shiraishi K , Hara I 
Published: January 2020

Abstract
An HPLC method [J. Biochem. (Tokyo) 91:1381, 1982] was used for evaluating serum lipoproteins, with on-line monitoring of either cholesterol or phospholipids. Five well-distinguished lipoprotein fractions were observed, based on their particle sizes. Serum of 15 normal persons, 12 subjects with various types of hyperlipidemia, 20 patients with various liver diseases, and two cases of familial LCAT deficiency were examined and the results compared with those by a sequential ultracentrifugal floatation technique. In the normal group, the amounts of fractions 2, 3, and 4 by the HPLC method correlated well with concentrations of the LDL, HDL2, and HDL3 fractions as measured by the ultracentrifugal method, respectively. In the hyperlipidemic group, similar good correlations were observed between fractions 1, 2, 3, 4 and chylomicrons + VLDL, LDL, HDL2, and HDL3 fractions, respectively. For those with liver diseases or LCAT deficiency, the corresponding fractions correlated less well, and characteristically the elution profile of lipoproteins in these groups showed heterogeneity of particle size within each lipoprotein density class, especially in LDL and HDL2.
Enzyme-linked immunosorbent assay for detection of antibodies to extractable nuclear antigens in systemic lupus erythematosus, with nylon as solid phase.
Authors: Ishaq M , Ali R 
Published: January 2020

Abstract
In this enzyme-linked immunosorbent assay (ELISA) for detection of antibodies against extractable nuclear antigens (ENA) in sera of patients with systemic lupus erythematosus (SLE), nylon is used as solid phase for antigen binding instead of the commonly used polystyrene surface. Optimal conditions for activation of the nylon beads, antigen coating, and other relevant factors have been investigated. We compared the incidence of anti-ENA antibodies in SLE, using chromogenic and fluorogenic enzyme substrates. Of SLE patients, 54% were positive for anti-ENA antibodies when chromogenic substrate was used as compared with 68% for fluorogenic substrate. Antibody activity against Sm and RNP antigens was distinguished on the basis of ribonuclease sensitivity of the RNP antigen. The method described offers advantages such as decreased background activity, increased surface area, facility for prolonged storage of antigen-coated solid phase, and miniaturization of the assay.
Creatine kinase and lactate dehydrogenase: stability of isoenzymes and their activity in stored human plasma and prostatic tissue extracts and effect of sample dilution.
Authors: Shain S A , Boesel R W , Klipper R W , Lancaster C M 
Published: January 2020

Abstract
We examined the effect of storing human plasma or extracts of prostate at -90 degrees C on the activity of creatine kinase and lactate dehydrogenase and isoenzyme distribution. Enzyme activities were unaltered during storage for as long as six weeks. If these preparations were thawed only once at 2 to 4 degrees C, they could be stored for as long as 165 days at -90 degrees C with no change in isoenzyme distribution. Inexplicably, apparent isoenzyme distribution of prostatic lactate dehydrogenase was sensitive to sample dilution, whereas the isoenzyme distribution of lactate dehydrogenase in plasma was not. Our observations emphasize the importance of validating details of analytical protocols that are to be used for quantification of new types of specimens.
Modified technique for periodic acid-Schiff staining of glycoproteins on agarose-film electrophoretograms.
Authors: Trivedi S M , Frondoza C G , Humphrey R L 
Published: January 2020

Abstract
We have modified the periodic acid-Schiff staining technique for glycoproteins for use with thin agarose-film electrophoresis membranes. With this procedure, carbohydrate-rich proteins can be detected with negligible background staining and insignificant staining of nonglycoproteins such as albumin and nonglycosylated Bence Jones proteins (free light chains). On the other hand, carbohydrate-rich M components of immunoglobulins M and A in serum and in cerebrospinal fluid from patients with plasma cell dyscrasia are readily detected. The technique is two- to threefold more sensitive than Ponceau S. Glycoproteins in serum and body fluids can be determined as a routine analytical procedure.
Clinical laboratory evaluation of Ames' Seralyzer Reflectance Photometer for total bilirubin and cholesterol.
Authors: Brown S , Savory J , Wills M R 
Published: January 2020

Abstract
We evaluated the dry-reagent technology for determining cholesterol and adult total bilirubin in serum with the Ames Seralyzer Reflectance Photometer. This instrument is designed to measure the intensity of light of a selected wavelength that is reflected from reagent-impregnated test strips saturated with sample. Regression analysis of the data for total bilirubin in adults (y) compared with corresponding values obtained with the Technicon SMAC (x) yielded the regression line: y = 1.07x + 2.5 (n = 128, r = 0.99). Day-to-day precision (CV) was 3.8% at 8.1 mg of bilirubin per liter, 5.34% at 48.3 mg/L. Regression analysis of the cholesterol data (y) vs cholesterol values obtained with an enzymatic cholesterol method (x) yielded the regression line: y = 0.91x + 191 mg/L (n = 100, r = 0.98). Day-to-day CVs for cholesterol were 3.6% at 1.62 g/L, 3.11% at 2.62 g/L.
Liquid-chromatographic measurement of creatinine in serum and urine.
Authors: Okuda T , Oie T , Nishida M 
Published: January 2020

Abstract
We describe the adaptation of a "high-performance" liquid chromatographic method for determination of creatinine in serum and urine. The proposed method is simple, rapid, precise, and accurate. The retention time for creatinine can be varied simply by changing the KH2PO4 concentration in the mobile phase: acetonitrile/aqueous KH2PO4 (1/4 by vol). Within-day precision (CV) was 1.2-3.6% in serum chromatographed with an internal standard, and 2.3-2.8% in serum when an external standard was used. Between-day precision (CV) was 1.3-2.1% in serum and 1.3-2.7% in urine (with an external standard). Analytical recoveries of creatinine added to serum were 94-100% for the method with an internal standard, 95-103% with an external standard.
Characteristics of high-affinity folate binding in human leukocytes.
Authors: Holm J , Hansen S I , Lyngbye J 
Published: January 2020

Abstract
High-affinity binding of [3H]folate in leukocytes from normal subjects was studied in equilibrium dialysis experiments (pH 7.4, 37 degrees C). Binding displayed positive cooperativity, and the binding affinity increased with decreasing concentration of the binding protein. Both phenomena could be interpreted in terms of ligand binding to a polymerizing system where the affinity of ligand for the oligomer is greater than its affinity for the polymer prevailing at higher concentrations of the binding protein.
Measurement of true salicylate concentrations in serum from patients with Reye's syndrome.
Authors: Kang E S , Todd T A , Capaci M T , Schwenzer K , Jabbour J T 
Published: January 2020

Abstract
Patients with Reye's syndrome who have been given aspirin are said to maintain higher-than-anticipated salicylate concentrations in blood, for longer than expected. We explored whether this could be attributed to spurious results from nonsalicylate compounds in the Trinder reaction for salicylates. All of 63 organic acids and amines examined that form colored complexes with Trinder's reagent had detectable absorbance at 540 nm at 0.2 g/L, including some endogenous compounds known to be increased in Reye's syndrome patients and many others endogenous in humans. By subjecting deproteinized sera to thin-layer chromatography and eluting the salicylate fraction before complexing it with ferric ion, true salicylate can be measured quantitatively and differentiated from interfering compounds. In addition, when we examined the effect of salicylate on palmitate binding to serum proteins, we found that salicylate concentrations of 0.2 g/L displaced [16-14C]palmitate binding to protein more in Reye's syndrome patients than in Reye's syndrome survivors or children with influenza. This suggests the presence of atypical binding characteristics for salicylate and palmitate in the acute disorder but not in survivors or children with influenza.
Enzyme inactivation in serum before determination of total bile acids.
Authors: Hanson N Q , Freier E F 
Published: January 2020

Abstract
Endogenous NADH-generating enzymes must be inactivated before total serum bile acids can be measured accurately by the direct enzymic method. To do this, we pretreat the sera with NaOH, in a final concentration of 0.1 mol/L. Consequently, lactate dehydrogenase activity at least as high as 30 000 U/L is destroyed, obviating blank determinations. Values for bile acid in serum, so obtained, agree with values obtained after pretreatment with heat, an alkali-methanol solution, or sodium pyruvate, but our pretreatment has the advantages of ease, speed, economy, and negligible blank values.
Reagent for the enzymatic determination of serum total cholesterol with improved lipolytic efficiency.
Authors: Siedel J , Hägele E O , Ziegenhorn J , Wahlefeld A W 
Published: January 2020

Abstract
We describe a sensitive method for quantifying the extent of cholesterol ester cleavage during enzymatic assay of total cholesterol in serum. Lipids are extracted from the assay mixture with chloroform/methanol (1/1 by vol), concentrated, then quantified by "high-performance" thin-layer chromatography. Although with conventional enzymatic reagents for determination of serum total cholesterol the hydrolysis of the cholesterol esters may be incomplete, a new enzymatic cholesterol reagent (Monotest Cholesterol, High Performance, Boehringer Mannheim) gives virtually complete cholesterol ester cleavage (i.e., greater than or equal to 99.5%). Use of this reagent with its improved lipolytic efficiency yields results for serum total cholesterol that are identical to those measured with a candidate reference procedure involving alkaline cholesterol ester saponification.
Glycosylated hemoglobin measured by affinity chromatography: micro-sample collection and room-temperature storage.
Authors: Little R R , England J D , Wiedmeyer H M , Goldstein D E 
Published: January 2020

Abstract
Under proper conditions, whole blood can be stored at room temperature for as long as 21 days before measurement of glycosylated hemoglobin by affinity chromatography. Whole blood (anticoagulated with EDTA or heparin) was placed in capillary tubes, which were then sealed at both ends and stored at room temperature. Just before assay, whole blood was rinsed from the tubes and diluted 10-fold with water. Samples of each patient's blood were assayed as whole-blood hemolysates by affinity chromatography after zero, seven, 14, and 21 days of storage. Values for glycosylated hemoglobin did not change over 21 days of storage and values for each storage day correlated well (r = 0.97, p less than .0001) with hemoglobin A1C measured in fresh erythrocyte hemolysates by "high-performance" liquid ion-exchange chromatography.
Liquid-chromatographic determination of aminoglutethimide in plasma.
Authors: Robinson B A , Cornell F N 
Published: January 2020

Abstract
A simple, rapid "high-performance" liquid-chromatographic procedure is presented for the determination of aminoglutethimide in plasma. After precipitation of the protein with acetonitrile, an aliquot of the supernate is injected directly onto a radially compressed, reversed-phase column. The aminoglutethimide is isocratically eluted with a mobile phase of acetonitrile/water/tert-butyl ammonium phosphate. The method is both accurate and precise and has been in routine use in our laboratory for more than 12 months.
Automated particle-counting immunoassay of C-reactive protein and its application to serum, cord serum, and cerebrospinal fluid samples.
Authors: Collet-Cassart D , Mareschal J C , Sindic C J , Tomasi J P , Masson P L 
Published: January 2020

Abstract
The assay for C-reactive protein has been fully automated as a particle-counting immunoassay. For cerebrospinal fluid, cord serum, and adults' serum its range of sensitivity extends from 1 microgram/L to 300 mg/L, with a minimal sample dilution of twofold and a maximal dilution of 50-fold being required. This range is so broad because free antibodies are added to the reaction medium. However, we have used Fab fragments rather than whole antibody to avoid too steep a standard curve and a decrease of agglutination at high concentrations of antigen. For 99 consecutive cord sera examined, the concentrations of C-reactive protein ranged from 7 micrograms/L to 1.750 mg/L. The geometric mean was 50 micrograms/L and the upper normal limit (geometric mean +/- 2 SD of the log values) was established at 525 micrograms/L.
Dipalmitoylphosphatidylcholine in amniotic fluid quantified by fast-atom-bombardment mass spectrometry.
Authors: Ho B C , Fenselau C , Hansen G , Larsen J , Daniel A 
Published: January 2020

Abstract
Dipalmitoylphosphatidylcholine (DPC) is quantified by taking advantage of stable-isotope-labeled d9-DPC as internal standard. Use of a mass spectrometer to measure the ratio of d9/d0 makes this procedure a quantitative one. d9-DPC was synthesized by refluxing dipalmitoylethanolamine with d3-methyl iodide in methanol in the presence of sodium bicarbonate for 26 h. The yield of d9-phosphatidylcholine (d9-lecithin) was 89% after column-chromatographic purification. Fast atom bombardment was used to desorb the preformed phosphatidylcholine ions in a mass spectrometer of Nier-Johnson geometry. In our assessment of accuracy and precision of this technique, we found a correlation coefficient of 0.9994 between signal and sample concentration. The method was less precise when the total d0- plus d9-DPC was less than 0.2 micrograms or when the ratio of d0- to d9-lecithin exceeded 100. The within-run CV was about 1.0%. The amount of DPC in amniotic fluid samples assessed by mass spectrometry was compared with results for total phosphatidylcholine quantified by thin-layer chromatography. The fate of DPC in various laboratory manipulations was also studied.
Detection of morphine in urine by hemagglutination inhibition, with use of lyophilized reagents.
Authors: Vanzetti G , Cassani M , Valente D 
Published: January 2020

Abstract
We describe a modified test of hemagglutination inhibition for the detection of morphine in urine, similar to the well-known test for pregnancy. The reaction takes place in test tubes or ampoules containing carefully matched amounts of lyophilized morphine antiserum and tanned human erythrocytes coated with morphine conjugated to rat serum albumin. The reagents are reconstituted by adding 100 microL of urine and 400 microL of water, and the result is read after 60 min. The detection limit, tested with the method of Gorodetzky (Clin Chem 19:753, 1973), was about 200 ng of total morphine per milliliter of urine. For more than 2000 samples, results by our test agreed satisfactorily with those obtained by an accepted RIA method. The test is suitable for rapid screening in field work, monitoring subjects during detoxication, and use in nonspecialized laboratories. Confirmatory analysis is needed for quantitative measurements, forensic purposes, and discrimination between morphine and cross-reacting opiates.
Radioimmunoassay for human corticosteroid-binding globulin.
Authors: Rosner W , Polimeni S , Khan M S 
Published: January 2020

Abstract
We isolated corticosteroid-binding globulin from human plasma and used the isolated protein as the antigen in raising antibodies in rabbits. The resulting antiserum was used to develop an RIA with which we can determine the globulin in as little as 0.007 microL of serum, although we routinely use, after appropriate dilution, 0.02 microL. The mean concentration of corticosteroid-binding globulin in the serum of 40 normal subjects was 32.4 (SD 4.2) mg/L. The correlation (r) between results with this RIA and those by an established binding assay was 0.94.
Effect of Fluosol-DA on radioimmunoassay results.
Authors: Hammarstrom M , Mullins R , Sgoutas D 
Published: January 2020

Abstract
Fluosol-DA, an emulsion of perfluorocarbons, is used as a blood substitute. Its presence in samples affects results of certain radioimmunoassays routinely done in our laboratory. In several, added Fluosol-DA caused an upward displacement of the standard curve, the extent of which depended on the structure and quantity of the assayed antigen, the concentration of Fluosol-DA, and the nature of the immunoreagents. Fluosol-DA markedly affected results of assays for triiodothyronine, thyroxin, and digoxin (by one method), but assays for aminoglycosides and protein hormones were unaffected. We determined that Fluosol-DA interacts with unbound labeled (and, presumably, with unlabeled) antigen to form a complex that coprecipitates with the bound antigen-antibody fraction during the separation step. In comparison with the idealized condition we observed an apparent increase in antibody-bound labeled antigen and consequently an apparent underestimation of the measured antigen. Depending on the nature of the immunoreagents, antibodies can extract more or less of their respective antigens from particles of Fluosol-DA emulsion.
Influence of age and sex on the concentration of free thyroxin in serum and on the free thyroxin: total thyroxin ratio.
Authors: Wilke T J 
Published: January 2020

Abstract
The influence of age and sex on the concentration of free thyroxin and on the free thyroxin/total thyroxin ratio was studied in 104 men and 89 women, ages 15 to 83 years. Free thyroxin gradually declined with age in men but not in women. In addition, sex- and age-related variations for free thyroxin were similar to those for total thyroxin and the two indirect assessments of free thyroxin: the free thyroxin index and the total thyroxin/thyroxin binding globulin ratio. Further, free thyroxin and the free thyroxin/total thyroxin ratio were higher in men than in women. However, the free thyroxin/total thyroxin ratio showed relatively little change with age in either sex. This study suggests that the free thyroxin/total thyroxin ratio is helpful in minimizing the age-related variations in both free and total thyroxin.
Enzyme immunoassay of thyroxin-binding globulin in dried blood samples on filter paper.
Authors: Hata N , Ito M , Mizuta H , Nose O , Miyai K 
Published: January 2020

Abstract
A double-antibody enzyme immunoassay was developed for determination of thyroxin-binding globulin in dried blood samples on filter paper. The measurable concentration range of thyroxin-binding globulin in two 3-mm blood discs was 3.3 to 52 mg/L equivalent of serum (i.e., equivalent to the concentrations in known serum standards). Thyroxin-binding globulin in dried blood samples on filter paper was stable for at least four weeks when kept dry at -20 degrees C, 4 degrees C, or room temperature. The mean coefficients of variation were 6.6% (within assay) and 5.9% (between assays). The concentrations of thyroxin-binding globulin in dried blood samples determined by this method correlated well with those in serum determined by radioimmunoassay (r = 0.95) and by enzyme immunoassay (r = 0.96). This method is applicable for detecting cases of thyroxin-binding globulin deficiency and avoids the false-positive results for neonatal hypothyroidism obtained by measuring thyroxin.
Unconjugated estriol in serum as measured by liquid chromatography with electrochemical detection, compared with radioimmunoassay.
Authors: Kaplan L A , Hohnadel D C 
Published: January 2020

Abstract
Concentrations of unconjugated estriol in maternal serum or plasma increase throughout pregnancy, particularly during the third trimester. We present and discuss the results of a comparative study of unconjugated estriol as measured by a new "high-performance" liquid-chromatographic assay and by a conventional semiautomated radioimmunoassay procedure. In the new method, serum is extracted and chromatographed on a reversed-phase mu Bondapak-C8 column under radial compression. The estriol peak is detected with a glassy-carbon electrochemical detector. The chromatographic results (y) correlated well with the RIA results (x) for 105 samples from 45 pregnant women in their third trimester (y = 1.07x - 0.55 micrograms/L, r = 0.933), with no significant difference between the means of the two methods.
Two-site immunochemiluminometric assay for human alpha 1-fetoprotein.
Authors: Weeks I , Campbell A K , Woodhead J S 
Published: January 2020

Abstract
In this two-site immunochemiluminometric assay for human alpha 1-fetoprotein, acridinium ester-labeled monoclonal antibodies are used that have an average incorporation ratio of 0.3 mol of acridinium ester per mole of antibody. The solid-phase antibody consists of sheep anti-alpha 1-fetoprotein IgG covalently coupled to a diazonium derivative of reprecipitated aminoaryl cellulose. The assay involves a 1-h incubation after the simultaneous incubation of labeled and solid-phase antibodies. The subsequent quantification time is 10 s per tube. The assay has a working range of 20 to 246 kilo-int. units/L for CVs less than or equal to 10% and a sensitivity of detection of 1.3 kilo-int. units/L. Experiments with monoclonal antibodies labeled to an average incorporation ratio of 2.8 mol of acridinium ester per mole of antibody suggest that assay sensitivity increases with increased specific activity. alpha 1-Fetoprotein concentrations in the sera of pregnant women at 14-20 weeks of gestation, as measured by the present assay, agreed with the results of a conventional radioimmunoassay.