bluuebunny/crossref_metadata_2025_split huggingface dataset:
Intracellular precursor forms of plasma proteins: their functions and possible occurrence in plasma.
Authors: Peters T 
Published: January 2020

Abstract
An attempt is made to review the steps in the biosynthesis of plasma proteins by the liver and to point out intermediate precursor forms that might escape to the plasma. With the use of immunochemical techniques such as immunofixation electrophoresis it is possible to identify these precursors when they present an unusual bands on protein electrophoresis. The discovery of the role of vitamin K in the formation of the blood-clotting proteins, for example, followed the immunochemical detection of an abnormal form of inactive prothrombin in plasma. The chief types of precursors likely to occur in blood are forms that are incompletely glycosylated, phosphorylated, or sulfated, or which are single peptide chains of multichain proteins. Precursors containing basic propeptides such as proinsulin or proalbumin may also appear. By being alert to the possible appearance of intracellular forms in plasma, the clinical chemist may be able to relate their presence to the nature of the disease process responsible.
Nonisotopic M13 probes for detecting the beta-globin gene: application to diagnosis of sickle cell anemia.
Authors: Sheldon E , Kellogg D E , Levenson C , Bloch W , Aldwin L , Birch D , Goodson R , Sheridan P , Horn G , Watson R 
Published: January 2020

Abstract
M13 DNA probes labeled with biotinylated psoralen and a streptavidin-horseradish peroxidase conjugate provide nonradioactive detection of the sickle cell and normal alleles of the beta-globin locus. The two biotinylated probes contain single-stranded sequences complementary to two different Sau3AI restriction fragments from the 5' region of the beta-globin gene and double-stranded M13 vector sequences. These probes are labeled with biotinylated psoralen photochemically linked to DNA. After hybridization, the presence of biotinylated probe bound to target DNA is detected in 3 h by using a streptavidin-horseradish peroxidase conjugate and the substrate, 3,3',5,5'-tetramethylbenzidine. Digestion of the normal (beta A) allele of the beta-globin gene with MstII (or isoschizomers) yields a 1.14-kb restriction fragment, while digestion of the mutant beta S allele yields a 1.34-kb fragment. These fragments can be resolved by gel electrophoresis and detected by Southern blot hybridization. The nonradioisotopic probe system can detect the beta-globin restriction fragment in as little as 0.5 microgram of human DNA and can distinguish heterozygotes (beta A beta S) from homozygotes (beta A beta A or beta S beta S) in 2.0 micrograms of human DNA.
Measurement of intact human parathyrin by an extracting two-site immunoradiometric assay.
Authors: Blind E , Schmidt-Gayk H , Armbruster F P , Stadler A 
Published: January 2020

Abstract
This is an immunoradiometric assay of intact human parathyrin, hPTH(1-84). One antibody, directed against the N-terminal part of the hormone, was produced in goats and conjugated covalently to cellulose particles. hPTH(1-84) and the N-terminal fragments were extracted from EDTA-treated plasma by these particles and thus concentrated. Another antibody, against synthetic hPTH(53-84), was raised in rabbits; this bound to the C-terminal part of the hormone. The final step was labeling the second free binding site of this antibody with 125I-labeled Tyr52-hPTH(53-84) and measuring the bound radioactivity. This assay can detect intact PTH in concentrations as low as 0.6 pmol/L (1.2 X 10(-16) mol per tube). The assay did not cross react with hPTH(1-34), hPTH(1-44), hPTH(28-48), hPTH(39-84), hPTH(44-68), or hPTH(53-84) in concentrations up to 6400 pmol/L. In 60 normal subjects, hPTH(1-84) concentrations ranged from 1.9 to 6.8 pmol/L; in 32 patients with primary hyperparathyroidism, from 7.0 to 80 pmol/L. The hormone was not detected in four patients with hypoparathyroidism.
"Delfia" and "Amerlite": two sensitive nonisotopic immunoassay systems for assay of thyrotropin compared.
Authors: Parnham A J , Tarbit I F 
Published: January 2020

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-batch precision over a wide range of thyrotropin concentrations. Results by the two methods correlate well (r = 0.992); the regression equation is: Amerlite = 0.915 Delfia - 0.33 milli-int. unit/L. The standard curve for the Delfia assay was linear, but that for the Amerlite assay showed some deviation from linearity below 0.5 milli-int. unit/L. Both assays have a negative bias in comparison with radiolabeled immunoradiometric assays, as judged by results for samples from the Quality Assurance Scheme. Both assays discriminate well between hyper-, hypo-, and euthyroid subjects, and results for thyrotropin for most patients with nonthyroidal illness were within the euthyroid reference interval. Both assays are convenient to perform and are based on systems that provide a viable alternative to radioimmunoassay.
Rapid typing of serum paraproteins by immunoblotting without antigen-excess artifacts.
Authors: Norden A G , Fulcher L M , Heys A D 
Published: January 2020

Abstract
In this new immunoblotting procedure for determining the heavy-chain class and light-chain type of monoclonal serum immunoglobulins, proteins are transferred from agarose electrophoretic gels to nitrocellulose by brief capillary blotting. Paraproteins transferred are detected with appropriate horseradish peroxidase-conjugated antisera to light chain and heavy chain. Examination of 121 serum specimens probably containing a paraprotein (as detected by protein staining) by immunoblotting and by immunofixation gave the same results for 116 specimens: paraproteins were typed in 103 specimens and their presence was excluded in 13. Immunoblotting required one repeat analysis (as compared with 22 for immunofixation). In only 100 min we could type as many as 10 paraproteins, and the procedure did not show antigen-excess artifacts. These results suggest that immunoblotting may be preferable to immunofixation for routine typing of paraproteins.
Deciding the optimum interval between specimen collections: theory and nomograms.
Authors: Fraser C G , Browning M C 
Published: January 2020

Abstract
The time interval between collection of specimens from an individual patient is usually determined empirically. For analytes whose values decline according to first-order kinetics--for example, enzyme activities in serum after an acute myocardial infarction, tumor markers in serum after excision of the tumor, and drugs in serum after an overdose--the minimum time between collections (delta T) depends on the elimination half-life (t) and the analytical precision (CVA), according to the equation: delta T = (1/log2) X t X log (2.33 CVA/100 + 1). Nomograms showing this relationship graphically have been generated.
Analytical systems ranked by freedom from interferences.
Authors: Glick M R , Ryder K W 
Published: January 2020

Abstract
We determined the effect of hemolysis, lipemia, and bilirubinemia on clinical-chemical analytical results under standardized conditions, for serum specimens prepared by us. Our purpose was to assess results obtained with 22 commonly available analytical systems. The quantitative rating scheme described is derived from the observed interference(s) divided by the number of analytical methods evaluated. The combined ranking reveals which chemistry analyzers are least affected and which are most affected by the added substances. Generally, systems that incorporate physical barriers or protein-separation steps perform better than those without thin-film layers, glass-fiber barriers, or dialysis membranes. Among the "direct" analyzers, fewer interferences are seen if appropriate "blanking" wavelengths are used, especially if a "specimen blank" absorbance is used appropriately in the analytical system. Centrifugal analyzers tended to perform poorly, according to the criteria presented here.
Cholinesterase assay automated in the Cobas-Bio centrifugal analyzer.
Authors: Ratnaike S , Gray F , Deam D 
Published: January 2020

Abstract
We describe a method for measuring plasma cholinesterase (EC 3.1.1.8) and dibucaine and fluoride numbers by using the Cobas-Bio. Benzoyl choline chloride is used as substrate. Reaction conditions are the same as in the corresponding manual method. The reaction is stopped with physostigmine. Choline oxidase (EC 1.1.3.17) is coupled with 4-aminoantipyrene. In the presence of peroxidase (EC 1.11.1.7), the 2-hydroxy-3,5-dichlorobenzenesulfonate indicator reaction gives a red product, measured at 505 nm. The analyzer is used in the "Multi Run" mode, with plasma cholinesterase and dibucaine and fluoride inhibition concurrently measured for eight samples. Coefficients of correlation between the manual and present method for plasma cholinesterase and dibucaine and fluoride numbers in 40 patients of various phenotypes were 0.843, 0.923, and 0.717, respectively. The inter-batch CV for each of the three assays was 2% to 3%.
Automated quantification of thyrotropin by radial partition immunoassay.
Authors: Rugg J A , Flaa C W , Dawson S R , Rigl C T , Leung K S , Evans S A 
Published: January 2020

Abstract
We describe a radial partition enzyme immunoassay in which fully automated quantification of human thyrotropin (hTSH) takes less than 11 min. This "sandwich"-type assay involves two monoclonal antibodies, both specific for the intact hTSH molecule. The solid phase consists of tabs of glass-fiber filter paper containing a pre-immobilized monoclonal anti-hTSH antibody complexed with a goat antibody specific for the Fc region of mouse IgG. The patient's sample is first applied to the central "reaction zone" of the tab, wherein hTSH binds to the immobilized antibody. Application of a buffered solution containing enzyme-labeled Fab' fragments of the second monoclonal anti-hTSH antibody initiates "sandwich" formation. A wash buffer containing a fluorogenic substrate elutes unbound conjugate to the tab periphery. The bound enzyme conjugate is quantified by measuring the rate of increase in fluorescence in the reaction zone of the tab, then converting the rate to clinical units by comparison with a stored calibration curve. The clinical utility and performance of the present assay compare favorably with those of other sensitive assays for hTSH.
Bias between enzymatic methods and the reference method for cholesterol.
Authors: Kroll M H , Lindsey H , Greene J , Sliva C , Hainline A , Elin R J 
Published: January 2020

Abstract
Assaying 312 serum samples, we compared four enzymatic methods for serum cholesterol with the Reference Method (modified Abell-Kendall) of the Centers for Disease Control (CDC). The means for the aca, TDx, and SMAC methods (2.27, 2.27, and 2.24 g/L, respectively) were significantly higher (P less than 0.05) than those of the Reference Method and the RA-1000 method (2.19 and 2.18 g/L, respectively). The biased methods had positive proportional and (or) systematic biases. Results with these methods were 2.6% to 4.9% higher than with the Reference Method. The assigned concentrations of cholesterol in the calibration materials for the SMAC and aca agreed with those obtained by the Reference Method, but were lower for the TDx and higher for the RA-1000. These findings document positive biases for cholesterol with three enzymatic methods and suggest that misassignment of calibrators is not primarily responsible for the biases found with the aca and SMAC. It may, however, to be a significant factor for the TDx.
Variations in the composition of low- and high-density lipoproteins during the acute phase of myocardial infarction.
Authors: Mainard F , Madec Y , Robinet N 
Published: January 2020

Abstract
We analyzed correlations between apolipoprotein B (apo B), cholesterol and phospholipids (preponderant lipids) in low-density lipoproteins (LDL) as well as between apolipoprotein A1 (apo A1) and these same lipids in high-density lipoproteins (HDL), during the acute phase of myocardial infarction. In LDL, a very elevated and stable correlation (r) was observed between these parameters, and the coefficients of regression (b) did not differ significantly during the period studied. In HDL, there was a decrease in r and b values from day 1 to day 2, then an increase after day 2. We hypothesize that these disturbances in HDL composition may be due to a greater endocytosis of LDL at day 2, leading to intracellular increase in cholesterol and phospholipids. Part of these lipids could be taken up by HDL molecules, causing a transient overload.
Four assay methods for glycated hemoglobin compared as screening tests for diabetes mellitus: the Islington Diabetes Survey.
Authors: Forrest R D , Jackson C A , Gould B J , Casburn-Budd M , Taylor J E , Yudkin J S 
Published: January 2020

Abstract
We assessed the utility of four methods of glycated hemoglobin assay (agar gel electrophoresis with Schiff base, agar gel electrophoresis with prior removal of Schiff base, boronate affinity chromatography, and isoelectric focusing) as screening tests for diabetes mellitus, studying 223 subjects undergoing an oral glucose-tolerance test after fasting and 2 h after ingestion of 75 g of glucose. Assessment of glucose tolerance status according to the 1980 World Health Organization diagnostic criteria indicated that 13 subjects (5.8%) had diabetes, 48 (21.5%) had impaired glucose tolerance, and nine (4.0%) could not be classified (six of these because of missing values). Use of receiver-operating characteristic curves to compare the assays as screening tests showed the affinity chromatography assay to be superior. Assays for glycated hemoglobin after fasting had better precision than post-glucose-load assays as screening tests, and test characteristics of glycated hemoglobin assays on fasting subjects were similar to those of the blood-glucose estimation after fasting. We conclude that measurement of glycated hemoglobin may be a useful screening test for diabetes, and that such measurement of stable glycated hemoglobin is as accurate as measurement of fasting blood-glucose in screening for diabetes. For this use, we found methods that measure stable glycated hemoglobin superior to that measuring both stable and labile Schiff-base fractions.
Galactosemia detection from phenylketonuria screening.
Authors: Henderson M J , Shapiro L , McCowan C 
Published: January 2020

Abstract
We describe a case of classical galactosemia in which the diagnosis was first suggested by the finding of a moderately increased blood-spot phenylalanine concentration. The child was clinically unaffected at six days when the initial sample was collected. Prompt institution of dietary management averted a serious metabolic crisis.
Gas-chromatographic measurement of carboxyhemoglobin in blood from mothers and newborns.
Authors: Fogh-Andersen N , Eriksen P S , Grinsted J , Siggaard-Andersen O 
Published: January 2020

Abstract
HbCO in blood sampled from 20 mothers and newborns immediately after birth was measured with a new, simple gas-chromatographic method for CO. The mean ratio of HbCO to total hemoglobin for 13 non-smoking mothers did not differ significantly from that for their infants (mean 0.38%, SD 0.26% vs 0.38%, SD 0.13%), but the HbCO ratio varied more in the mothers than in the infants (P less than 0.05). The infants of seven cigarette-smoking mothers, tobacco-abstinent for 7 h during labor, had higher HbCO ratios than their mothers (mean 1.88% vs 1.28%, P less than 0.05). The results are in harmony with the concept of equal affinities of fetal and adult hemoglobin for CO and a long time for passage of CO across the placenta.
Liquid-chromatographic measurement of cyclosporin A and its metabolites in blood, bile, and urine.
Authors: Christians U , Zimmer K O , Wonigeit K , Maurer G , Sewing K F 
Published: January 2020

Abstract
Using solid-phase extraction columns and "high-performance" liquid-chromatographic (HPLC) analysis, we could determine cyclosporin A and nine of its metabolites in blood, bile, and urine. To facilitate calculations of concentrations of cyclosporin A and its metabolites from the chromatograms, we used cyclosporin D as internal standard. For the HPLC analysis we used two sequential 250-mm analytical columns filled with reversed-phase octyl (C8) sorbent, eluting with a concave gradient of water, adjusted to pH 3.0 with phosphoric acid, and acetonitrile. Peaks were detected at 205 nm. For characterization of the chromatographic peaks, we isolated, by semi-preparative HPLC, 32 fractions representing peaks potentially related to cyclosporin A metabolites and re-injected them into the HPLC system under the same conditions as authentic cyclosporin A metabolites. Analytical recovery was 70-80%. The inter-assay CV for bile was 7.2%, for urine 12.3%. The method was used for routine monitoring of cyclosporin A and its metabolites.
Direct spectrophotometry of bilirubin in serum of the newborn, with use of caffeine reagent.
Authors: Vink K L , Schuurman W , van Gansewinkel R 
Published: January 2020

Abstract
We recently described a direct spectrophotometric method for unconjugated bilirubin, with caffeine reagent (Clin Chem 1986;32:1389-93). Because this method is independent of the protein matrix we used it for the preparation of bilirubin standards (Clin Chem 1987;33:1817-21). Now, in this paper, we utilize the caffeine reagent in setting up a bilirubin method for serum from neonates. This resulted in a two-wavelength (465 and 528 nm) equation, which fully corrects for HbO2 interferences. In combination with a bilirubin standard, this equation may be transformed into a simple relative formula for use with this simple dilution method. We studied this two-wavelength method with 55 neonates' sera, comparing results with those by both the diazo method of Doumas et al. (Clin Chem 1985;31:1779-89) and the borate method of Hertz et al. (Scand J Clin Lab Invest 1974;33:215-30). We found that this new method is independent of hemolysis and of the matrix of the sera. Therefore, it is very suitable for use in neonatology.
Highly specific enzyme immunoassay for the beta-subunit of human thyrotropin with use of monoclonal antibodies.
Authors: Iijima Y , Endo Y , Hata N , Fujita H , Unoki M , Miyal K 
Published: January 2020

Abstract
A highly specific enzyme-linked "sandwich" immunoassay is described for determining free human thyrotropin (hTSH) beta-subunit in serum by using a anti-hTSH beta-subunit monoclonal antibody conjugated with beta-D-galactosidase (EC 3.2.1.23) and a solid phase consisting of silicone rods coated with another monoclonal antibody. We could detect as little as 0.04 ng of beta-subunit per assay. The measurable range of hTSH beta-subunit concentrations in serum was 0.4 to 50 micrograms/L. The assay demonstrated little or no cross reactivity with intact hTSH, hTSH alpha-subunit, or human choriogonadotropin. The mean CVs were 12.2% within assay, 13.9% between assay. The hTSH beta-subunit was not detectable in sera from healthy subjects, patients with hyperthyroidism, or two patients with pituitary tumors producing TSH. It was measurable (at concentrations of 0.65 to 2.70 micrograms/L) in sera from eight of 23 hypothyroid patients. In five of the hypothyroid patients examined, the concentration of hTSH beta-subunit in serum increased after administration of thyroliberin. This method may be useful in elucidating the physiological and pathological significance of the hTSH beta-subunit and in examining the function of the hypothalamus-pituitary-thyroid axis.
An automated approach to characterize, in simple kinetic terms, the generation and inactivation of thrombin and the interaction of fibrinogen with thrombin.
Authors: Hoak D R , Banerjee S K , Kaldor G 
Published: January 2020

Abstract
Here, we used a fully automated, computer-directed centrifugal analyzer (which permitted simultaneous turbidimetry and calculation of results) and purified thrombin, fibrinogen, and various inhibitors to study clot formation. The Km and Vm for these reactions were useful in detecting and partly characterizing anticoagulants. We also explored the generation and inactivation of thrombin, using the two-stage prothrombin time and antithrombin activity tests. The amount of thrombin instantaneously generated and inactivated was monitored under artificially created pathological conditions. The pseudo-first-order rate constant for thrombin generation and inactivation and the instantaneous concentration of enzymatically active and inactive thrombin were used in the characterization of these conditions. We believe this approach is suitable for routine clinical use.
Non-resolving jaundice: bilirubin covalently attached to serum albumin circulates with the same metabolic half-life as albumin.
Authors: Reed R G , Davidson L K , Burrington C M , Peters T 
Published: January 2020

Abstract
In hepatobiliary disease and biliary obstruction, bilirubin often becomes covalently bound to albumin circulating in serum, producing a nondissociable complex. To determine how long this complexed bilirubin remains in the circulation, we compared the metabolic clearance of bilirubin-albumin complexes with the clearances of free bilirubin and unmodified albumin. Radiolabeled bilirubin, albumin, and covalent bilirubin-albumin were injected into the circulation of Sprague-Dawley rats and serial samples of plasma were analyzed for the injected compounds. The half-life of bilirubin was 6.2 min. The half-life of bilirubin covalently bound to rat serum albumin was 1.9 to 2.1 days, identical to that of unmodified rat albumin. We conclude that bilirubin covalently attached to albumin is maintained in the circulation with the long half-life of albumin rather than the short half-life of bilirubin. Because albumin in humans has a half-life of 19 days, covalent attachment of bilirubin to human albumin could result in persistence of hyperbilirubinemia long after the resolution of disease.
Some thermodynamic parameters and temperature-conversion factors for determining alpha-amylase concentration in serum.
Authors: Hafkenscheid J C , Hessels M 
Published: January 2020

Abstract
We measured the relationship between measured activity concentration and temperature for alpha-amylase (EC 3.2.1.1), using 10 different substrates. At 25-37 degrees C the Arrhenius plot was linear. The activation energy ranged from 33.3 kJ.mol-1 with maltoheptaose as substrate to 54.4 kJ.mol-1 with the Blue-Starch method. Activation energy was lowest for substrates having seven glucose moieties, the ones most suitable for determining alpha-amylase--i.e., the presence of more or fewer glycosyl units increased the activation energy. Substrates with blocked groups at the nonreducing end of the oligosaccharide chain were not considered here, because the relative reaction rates obtained with these substrates were less than those obtained with nonblocked substrates. We also determined temperature-conversion factors for alpha-amylase reaction with the various substrates. The results are discussed in relation to thermodynamic parameters of some other enzymes, e.g., creatine kinase and alkaline phosphatase.