bluuebunny/crossref_metadata_2025_split huggingface dataset:
Demonstration of alpha 1-acid glycoprotein (orosomucoid) by double one-dimensional slab gel electrophoresis: evidence for intra- and interindividual variability of the microheterogeneity pattern in health and disease.
Authors: Altland K , Roeder T , Jakin H M , Zimmer H G , Neuhoff V 
Published: January 2020

Abstract
This double one-dimensional slab gel electrophoresis technique, in the sequence polyacrylamide gel electrophoresis followed by isoelectric focusing in polyacrylamide gels, permits the selective demonstration and comparison of the micro-heterogeneity pattern of alpha 1-acid glycoprotein (orosomucoid) in as many as 96 human plasma or serum samples on one gel. The electrophoretic analysis is performed on 8 microL of serum or plasma without prior purification. Several hundred samples can be analyzed by one investigator during a working day. Densitometric evaluation of the patterns revealed two new findings: 1) The microheterogeneity pattern can be described by a parameter that is independent of the concentration of orosomucoid in plasma: the center of density of the pattern, with the peak number as coordinate. 2) There is a considerable average shift of the center of density towards more basic components among the patterns obtained for samples from hospital patients. The results suggest that followup of the intra-individual variation of the orosomucoid pattern in health and disease might help in studying the still-uncertain function of this protein as well as the diseases affecting the pattern.
Two-dimensional analysis of human lymphocyte proteins. III. Preliminary report on a marker for the early detection and diagnosis of infectious mononucleosis.
Authors: Willard K E 
Published: January 2020

Abstract
Two-dimensional gel electrophoretic patterns of human peripheral blood leukocytes from 12 patients with infectious mononucleosis were prepared by use of the ISO-DALT system. Before the two-dimensional separation, the leukocytes were purified by Ficoll-Paque gradient centrifugation and labeled overnight with [35S]methionine. Quantitative increases in two proteins were detected in the patterns of infected leukocytes from the patients as compared with controls. Fluorescence-activated cell sorting of leukocytes from normal human peripheral blood before subsequent two-dimensional gel analysis revealed that the dramatic increase in one of these proteins (Inmono:2) could be due to shifts in the population ratios of lymphocytes, monocytes, and granulocytes (Willard et al., manuscript in preparation). In contrast, the appearance of the infected leukocytes of a second protein, Inmono:1, could not be accounted for by cell-population shifts. Increased amounts of these two proteins have been found in every patient studied who had clinically detectable infectious mononucleosis. In addition, a patient who displayed symptoms of infectious mononucleosis but who did not have a positive result in the MONOSPOT Test (Ortho) until three weeks after our analysis also demonstrated increased relative amounts of these proteins in his leukocyte pattern.
Development and evaluation of a new solid-phase direct immunoenzyme assay for prostatic acid phosphatase.
Authors: Gericke K , Kohse K P , Pfleiderer G , Flüchter S H , Bichler K H 
Published: January 2020

Abstract
In this assay we used polystyrene-tube-attached rabbit antibodies against prostatic acid phosphatase (PAP) that had been purified to homogeneity from human prostate. The amount of immunoreactive acid phosphatase was determined directly by its enzymic activity in the solid-phase-bound immune complex. The detection limit was 0.05 U/L (0.13 microgram/L), the CVs between 4.3 and 10.8%. Investigating the organ specificity of PAP, we found that some cross-reacting acid phosphatase activity could be so measured in human kidney, leukocytes, and platelets, all of which probably contribute to the circulating "prostatic" acid phosphatase that normally is present in serum. Diurnal and day-to-day variations in serum PAP activity were as much as 100% in healthy subjects. Individuals without prostatic diseases (n = 92) had values for serum PAP activity up to 0.36 U/L (0.94 microgram/L), in an age-independent distribution; patients with benign prostatic hyperplasia (n = 62) showed values up to 0.48 U/L (1.25 micrograms/L). With PAP activity of 0.38 U/L or 1.0 microgram/L (90th percentile of the prostatic group) as the upper limit of "normality," overall sensitivity (stages A-D) for detection of prostatic cancer in 33 essentially untreated patients was 65%. Examples for the followup of therapy of prostatic cancer by measurement of serum PAP with this assay are described.
Enzyme immunoassay for free thyroxin.
Authors: Weetall H H , Hertl W , Ward F B , Hersh L S 
Published: January 2020

Abstract
We describe a mathematical model for a single-tube enzyme immunoassay for free thyroxin (FT4), involving use of a thyroxin/horseradish peroxidase (EC 1.11.1.7) conjugate that does not interact with thyroxin-binding globulin. In the presence of serum two populations of unassociated, or free, immunologically active constituents are present: FT4 and the conjugate. The concentrations of the former are determined by the serum constituents and of the latter by the albumin concentration. When a small quantity of antibody is added, it reacts with the variable amount of FT4 and with the constant amount of the conjugate, thus giving a measure of the FT4. We constructed a mathematical model based on thermodynamic binding constants and adsorption data. The model gives satisfactory agreement with the experimental data under a variety of experimental conditions. Results for 19 patients' serum samples demonstrate the validity of the concept.
Partition affinity ligand assay (PALA) for quantitation of triiodothyronine in serum.
Authors: Mattiasson B , Eriksson H 
Published: January 2020

Abstract
Radioimmunoassay of triiodothyronine was performed with a homogeneous incubation system. Bound and free ligand were separated by partitioning in an aqueous two-phase system. The agreement between the results obtained with this technique and conventional heterogeneous radioimmunoassay was very good (r = 0.981).
Incidence of gamma-globulin banding in a healthy population by high-resolution electrophoresis.
Authors: Papadopoulos N M , Elin R J , Wilson D M 
Published: January 2020

Abstract
We report the incidence of electrophoretically homogeneous protein bands occurring in the gamma-globulin region in serum samples from 600 healthy adults, as determined with a high-resolution electrophoretic system. One or more electrophoretically homogeneous protein bands of various intensities were detected in the gamma-globulin region of 30 specimens when we used an improved agarose electrophoretic system, and in two specimens when we used a cellulose acetate electrophoretic system. Improvement of the agarose gel electrophoretic system thus enhanced its sensitivity in defining homogeneous protein bands in the gamma-globulin region. The incidence of these bands in a healthy population is approximately 5%.
Some extensions to the GELLAB two-dimensional electrophoretic gel analysis system.
Authors: Lemkin P F , Lipkin L E , Lester E P 
Published: January 2020

Abstract
Several important extensions to the GELLAB 2D electrophoresis gel analysis system are discussed, including concepts that lead to the generation of consistent gel maps for a particular cell line, as illustrated by some recent results for the P388D1 cell system, GELLAB is an interactive graphics computer system that facilitates (a) image accessioning, (b) spot-data extraction from the image with position and density quantitation, (c) pairing of corresponding spots between gels, and (d) spot analysis for a large number of corresponding spots (over 3000 per gel) for up to 128 gels. Multiple gels are accessioned into the system used to create a composite gel data base. These then may be partitioned into multiple classes of gels and searched for statistically significant interclass spot differences, which can be visualized in reference map images of selected spot sets as well as mosaic images of particular spots in all gels. In addition to the visual feedback that is available during the gel analysis, various statistical, numerical, and other displays may be generated in the CGELP interactive program of the GELLAB system.
Automated assay for alpha 1-antitrypsin with N-alpha-benzoyl-DL-arginine-p-nitroanilide as trypsin substrate and standardized with p-nitrophenyl-p'-guanidinobenzoate as titrant for trypsin active sites.
Authors: Eckfeldt J H , Light R T , Leiendecker-Foster C 
Published: January 2020

Abstract
alpha 1-Antitrypsin is the most abundant of several serum protease inhibitors. Its deficiency is associated with an increased incidence of emphysema in adults, jaundice in newborns, and childhood cirrhosis. We describe an automated functional assay for the Instrumentation Laboratory's Multistat III Microcentrifugal Analyzer with N-alpha-benzoyl-DL-arginine-p-nitroanilide as trypsin substrate. The assay is standardized in terms of moles of trypsin active sites inhibited per liter of serum, by use of a chromogenic titrant for trypsin active sites, p-nitrophenyl-p'-guanidinobenzoate. The method is rapid, precise, and independent of trypsin supplier, and results correlate well with those by a manual chromogenic and a nephelometric assay.
Galactosyltransferase variant in pleural effusion.
Authors: Kim Y D , Weber G F , Tomita J T , Hirata A A 
Published: January 2020

Abstract
In measuring total galactosyltransferase activity in the pleural effusions from patients with benign or malignant diseases, we found no significant difference between the two groups (p greater than 0.05). However, a small amount of a galactosyltransferase variant, GT(l), could be separated from other galactosyltransferase enzymes in malignant pleural effusions by anion-exchange chromatography (DEAE-cellulose) with a buffer of low ionic strength. Other galactosyltransferases were eluted from the column with buffer of higher ionic strength. Using a mini-column method, we detected GT(l) enzyme in 19 of 26 specimens fro cancer patients, as compared with eight of 25 specimens from patients with benign disorders. The appearance of GT(l) enzyme in pleural effusion may be a tumor-associated phenomenon.
Influence of triglyceride-rich lipoproteins on nephelometry of apolipoprotein A-I.
Authors: Funke H , Assmann G 
Published: January 2020

Abstract
Measurement of apolipoprotein A-I in native human sera by endpoint or rate nephelometry may give falsely high results for sera that have a high triglyceride content. Such erroneous determinations of apolipoprotein A-I can be avoided if one instead assays supernates of sera from which apolipoprotein B-containing particles have been precipitated with phosphotungstic acid and MgCl2. A high triglyceride concentration in serum does not impair the completeness of the precipitation of apolipoprotein B-containing lipoproteins or the analytical recovery of apolipoprotein A-I. In a prospective epidemiologic study, by using rate nephelometry we found that there were different mean concentrations for apolipoprotein A-I in supernates of sera from men (1.36 g/L) and women (1.49 g/L). Furthermore, the known risk factors for atherosclerosis showed different correlations with apolipoprotein A-I than with high-density lipoprotein cholesterol.
Diagnostic and prognostic value of laboratory tests assessed in a follow-up study of 200 patients with liver disease.
Authors: Skrede S , Solberg H E , Ritland S , Blomhoff J P , Schrumpf E , Elgjo K , Gjone E 
Published: January 2020

Abstract
Two hundred patients with liver disease were re-studied after six to eight years. The initial routine diagnosis (mainly based on morphological criteria) was confirmed in almost 80% of cases of chronic liver disease. In chronic active hepatitis, agreement was found in 86%. An alternative classification based on cluster analysis of clinical chemical results from the first admission was also to a great extent confirmed by the follow-up study. Discriminant analysis of clinical chemical data correctly allocated 94% of patients with alcoholic cirrhosis (who had low haptoglobin and albumin) into groups with different prognosis. Clinical chemical results differed between verified and non-verified cases of primary biliary cirrhosis. By retrospective examination , we found that 15 of 16 patients with this disease or chronic active hepatitis who were initially misclassified by biopsy findings could be correctly diagnosed by discriminant analysis of clinical chemical results.
Retention of carcinoembryonic antigen on Sephadex G-50 and acrylamide gel buffer-exchange columns.
Authors: Haagensen D E , Sawlivich W , Davis S , Zamcheck N 
Published: January 2020

Abstract
We compared the degree of retention of carcinoembryonic antigen on two types of buffer-exchange columns, Sephadex G-50 and acrylamide gel. Both types retained trace amounts of the antigen applied in the form of a perchloric acid extract, as measured by its release into plasma samples subsequently processed through the same columns, Cross contamination, resulting in values falsely high by greater than 1 microgram/L, was observed when columns were re-used after processing plasma samples with carcinoembryonic antigen concentrations greater than 700 micrograms/L.
Commercial radioimmunoassay kit for measurement of alpha-fetoprotein adapted for use with dried blood specimens from newborns.
Authors: Mizejewski G J , Bellisario R , Beblowski D W , Carter T P 
Published: January 2020

Abstract
We adapted a commercial RIA kit to measure alpha-fetoprotein (AFP) in 0.785-microliters portions of 60-microliters spots of dried blood from newborns. We evaluated sample elution, temperature and time stability, between- and within-assay variability, sensitivity, and use of liquid vs dried specimens of blood. Also, we present normal AFP concentrations for healthy neonates during the first postnatal week. Our procedure permits measurement of AFP concentrations both in maternal liquid plasma and in spots of dried blood from the infant with the same RIA kit reagents and standards. The sensitivity, precision, stability, and simplicity of this procedure makes more practical the routine measurement of AFP in dried blood specimens from newborns than measurement in liquid plasma or serum. Blood-sample collection by heel stick suffices for this simple, efficient, inexpensive determination of AFP concentration in the newborn.
Improved nonradioactive method for routine clinical analysis for long- and medium-chain stool lipids.
Authors: Bandi Z L , Kenny M 
Published: January 2020

Abstract
We describe a simple, practical method for routine clinical determination of long- and medium-chain stool lipids. Stool lipids were extracted into a hot mixture of benzene/ethanol/water (12/59/29, by vol), saponified in 30 min without formation of ethyl esters, and titrated in a clear, one-phase medium to a sharp color change. CVs for the titration were 2, 1, and 1% for 0.05, 0.25, and 0.5 mmol of fatty acids, respectively. Interference of stool short-chain acids other than fatty acids was not statistically significant. The precision of sampling of stool homogenate was improved by analysis of two aliquots, one from the top and one from the bottom of the suspension. This method of sampling is a practical alternative to the unpleasant homogenization with a blender or to the addition of large quantities of bile salts. The accuracy of the method may be increased, in special cases, by removing potential interferants.
Emulsions of perfluorinated compounds for use in quality control of blood gas analyses.
Authors: Rej R , Schröder J 
Published: January 2020

Abstract
We prepared emulsions of perfluorinated compounds and examined their utility as liquid matrixes for tonometry in quality control of blood pH and gas analyses. Emulsions of perfluorotributylamine and perfluorodecalin, prepared by sonication, consisted largely of particles not exceeding 0.2 micrometer in diameter. Unlike most aqueous solutions these materials were not easily contaminated by room air and can be reliably used for tonometry liquids. Results for pCO2 and pO2 agreed well with calculated values. Average day-to-day coefficients of variation for emulsions of perfluorotributylamine were 2.9% for pH (within the range 7.11-7.60), 4.4% for pCO2 (2.88-9.17 kPa), and 1.6% for pO2 (7.76-40.67 kPa).
Comparison of the effectiveness of four clinical chemical assays in classifying patients with chest pain.
Authors: Van Steirteghem A C , Zweig M H , Robertson E A , Bernard R M , Putzeys G A , Bieva C J 
Published: January 2020

Abstract
We compared the usefulness of four serum assays for classifying patients originally suspected of having an acute myocardial infarction. One of these is the long-used measurement of total creatine kinase (CK) activity. The other three are relatively new immunoassays: myoglobin by RIA, CK-BB by RIA, and CK-MB by immunoinhibition. When we evaluated test effectiveness with use of conventionally derived reference ranges, the results were misleading. However, by using receiver operating characteristic curves, we were able to effectively compare the four tests at all possible decision levels, rather than at only one. Multiple closely sequential serum specimens were obtained during the first four days after the onset of chest pain. Total CK, CK-MB, and CK-BB all behaved similarly, reaching peak diagnostic effectiveness at 18-20 h, when all three correctly classified 95% of the infarct patients, with a zero false-positive rate. However, total CK was more useful in identifying infarcts later in their courses than were the two CK isoenzyme tests. Myoglobin assay was most effective earlier in the course, at about 7 to 8 h. Our results indicate (a) that the tests for myoglobin and for CK or its isoenzymes are complementary and (b) that of the three CK tests, measurement of total CK activity provides the most information over the broadest segment of a patient's course.
Free-cortisol assay by immunoextraction: comparison with an equilibrium dialysis procedure.
Authors: Clerico A , Del Chicca M G , Zucchelli G , Bartolomei C , Riccioni N 
Published: January 2020

Abstract
We describe a new method for directly determining the apparent free cortisol concentration in plasma samples by use of an antibody-coated test-tube RIA. Buffer-diluted plasma or standard serum is added to antibody-coated test tubes, incubated at 4 degrees C for 4 h, and the solutions are aspirated. 125I-labeled cortisol is added to each tube and incubated for 3 h at 4 degrees C. Then the insides of the tubes are washed and their radioactivities counted. The standard curve is in terms of free cortisol, the standard serum solutions having been measured with an equilibrium dialysis procedure. Plasma samples (n = 155) from normal subjects and from various patients, measured with the new immunoextraction method and the equilibrium dialysis technique, gave results that correlated well (r = 0.847, p less than 0.001). Results by this direct RIA also correlate well with the clinical adrenocortical status of patients for whom data on total plasma cortisol may be misleading. This simple, easy RIA is suited to be the routine method for free cortisol in plasma.
Rate nephelometry of human IgE in serum.
Authors: Beck O E , Kaiser P E 
Published: January 2020

Abstract
We have developed an immunoassay method for the determination of human IgE concentration in serum by rate nephelometry. Using the Beckman Immunochemistry Analyzer, human IgE antisera, and a human IgE standard, we analyzed 150 serum samples from normal and allergic patients. We report these results and those from comparison studies involving PRIST and Phadebas kits (both from Pharmacia), and Enzygnost (Behringwerke) kits. We also discuss diagnosis of allergy patients and their classification according to their concentrations of IgE and IgG4.
Immunoturbidimetry of albumin and immunoglobulin G in urine.
Authors: Teppo A M 
Published: January 2020

Abstract
I describe a rapid, sensitive immunoturbidimetric assay for measuring urinary albumin and immunoglobulin G with use of an automated spectrophotometer. Diluted urine samples and polyethylene glycol in phosphate-buffered saline are pipetted into the cuvettes of the spectrophotometer. The initial absorbances of the samples are measured at 340 nm; antiserum to albumin or to immunoglobulin G is added to each tube, and after 2 min at 37 degrees C the absorbance of the mixtures is read at 340 nm. The initial blank absorbances of the samples are subtracted from the final absorbances automatically. The change in absorbance is linear with concentration in the range of 5-400 mg/L for albumin and 3-1000 mg/L for IgG. The lower limit of the determination is 5 mg/L for albumin, 3 mg/L for IgG. Linear correlations were observed between the concentrations of albumin and immunoglobulin G determined by this method (x) and those determined by radial immunodiffusion (y). The regression equation for albumin was y = 0.84x + 0.03 (r = 0.99, n = 87), and for IgG y = 0.94x + 0.02 (r = 0.98, n = 87).
Direct determination of lipoprotein cholesterol distribution with micro-scale affinity chromatography columns.
Authors: Bentzen C L , Acuff K J , Marechal B , Rosenthal M A , Volk M E 
Published: January 2020

Abstract
A quick and practical procedure based upon the principles of affinity chromatography has been developed and specifically adopted for the separation of serum lipoproteins into their respective alpha- and beta-lipoprotein fractions. The cholesterol, phospholipids, apoproteins, and triglycerides of these two lipoprotein fractions--the high-density lipoproteins (HDL) and lower density lipoproteins--can be directly measured independently after this separation. The sums of each fraction agree with total serum components when independently assayed. The affinity column is packed with heparin bound in high capacity to agarose. The beta-lipoproteins (low-density and very-low-density lipoproteins) and HDL-containing apolipoprotein E are absorbed to the column support; the alpha-lipoproteins (non-apolipoprotein E) pass through. The beta-lipoproteins are subsequently desorbed from the column support with saline. The direct assay of both the "protective" alpha-lipoprotein cholesterol (HDL cholesterol) and of the presumably atherogenic lower-density beta-lipoprotein cholesterol (LDL + VLDL cholesterol) permits calculation of a beta-: alpha-lipoprotein cholesterol ratio, which may better indicate a patient's risk of stroke or coronary heart disease than does the value for HDL cholesterol alone.
Enzymic assay of creatinine in serum and urine with creatinine iminohydrolase and glutamate dehydrogenase.
Authors: Tanganelli E , Prencipe L , Bassi D , Cambiaghi S , Murador E 
Published: January 2020

Abstract
We describe an assay for creatinine in which it is converted by creatinine iminohydrolase (EC 3.5.4.21) into ammonia and N-methylhydantoin. The ammonia is subsequently assayed by use of alpha-ketoglutarate and glutamate dehydrogenase (EC 1.4.1.3). Use of NADPH as coenzyme eliminates all interferences from endogenous reactions. Endogenous ammonia in the sample is eliminated during a preincubation. The reaction reaches the endpoint in 15 min at working temperatures of 20-37 degrees C. No sample blank or reagent blank is needed. The standard curve is linear at least to 884 mumol (100 mg) of creatinine per liter. Average analytical recovery of creatinine in serum and urine is 99%. Within-run and between-run CVs are less than or equal to 2% and less than or equal to 6% for creatinine values of 335 mumol/L (38 mg/L) and 80 mumol/L (0 mg/L), respectively. Results by the described method (y) compare well with those by Jaffé's kinetic test (y = 1.01x -- 12.8), Berthelot/AutoAnalyzer method after treatment with immobilized creatinine iminohydrolase (y = 0.987x -- 13.2), Jaffé's test run on the SMA 12/60 (y = 1.011x -- 5.8), the Wahlefeld method (y = 1.014x -- 0.88), and Jaffé's test after deproteinization and absorption on fuller's earth (y = 0.985x -- 3.08). The method may be suitable for discrete, including centrifugal, automation.