bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development and comparative evaluation of immunoblot assays for detecting autoantibodies to Scl 70 and Jo 1 antigens in serum
Authors: Fonong T , Evans S M , Homburger H A 
Published: January 2020

Abstract
We developed rapid 24-h immunoblot assays for detecting autoantibodies to Scl 70 and Jo 1 antigens in serum. In comparative studies, we evaluated the analytical sensitivity of the immunoblot assays and commercial immunodiffusion assays for anti-Scl 70 and anti-Jo 1 autoantibodies with the use of positive control sera, and compared the frequencies of positive and negative results in a group of 116 sera, including specimens from 34 healthy controls and 82 patients with various connective-tissue diseases. The immunoblot assays were greater than 100-fold more sensitive than immunodiffusion for detecting both autoantibodies. Despite greater analytical sensitivity, there were no false-positive results by the immunoblot assay for anti-Scl 70 or anti-Jo 1 autoantibodies in sera from either the controls or the patients. The diagnostic sensitivity of the immunoblot assay for anti-Scl 70 autoantibodies in patients with scleroderma was greater than that of the immunodiffusion assay, 70% vs 20%, and was equivalent in patients with polymyositis, 43%. We conclude that rapid immunoblot assays for anti-Scl 70 and anti-Jo 1 autoantibodies are superior to immunodiffusion assays for clinical use and are suitable for routine use in the clinical laboratory.
Automation of specific human gene detection
Authors: Mayrand P E , Hoff L B , McBride L J , Bridgham J A , Cathcart R , Corcoran K P , Golda G S , Keith D H , Lachenmeier E W , Madden D E 
Published: January 2020

Abstract
An instrument/chemistry system is described that automates a new chemical procedure functionally equivalent to Southern blotting. A fluorescence gel scanner that detects migrating DNA fragments in real-time analyzes the samples produced by a prototype liquid-handling instrument that automates a solution-phase hybridization/solid-phase capture chemistry for DNA analysis. The combination of this chemistry, the gel scanner, and robotic automation eliminates the tedium encountered in traditional manual methods for specific gene detection and reduces analysis time from days to hours. Restriction fragment lengths are measured with high precision by comparison with in-lane standards to minimize effects attributable to migration anomalies. The utility of this automated system is demonstrated by executing a clinical research application involving hybridization to a multi-copy repeat sequence on the Y chromosome and its detection.
Apolipoprotein E polymorphism determined by restriction enzyme analysis of DNA amplified by polymerase chain reaction: convenient alternative to phenotyping by isoelectric focusing
Authors: Kontula K , Aalto-Setälä K , Kuusi T , Hämäläinen L , Syvänen A C 
Published: January 2020

Abstract
Three common alleles determine six apolipoprotein E (apo E) phenotypes that are associated with variations in serum cholesterol in the population. This genetic variation results from single nucleotide alterations at two DNA loci encoding the amino acid residues 112 and 158 of apo E. We compared results of apo E phenotyping carried out by isoelectric focusing with those of apo E genotyping accomplished by direct DNA analysis. In the latter, the target DNA was amplified by the polymerase chain reaction (PCR) and subsequently analyzed by digestion with the restriction enzyme Hha I, followed by polyacrylamide gel electrophoresis of the cleavage products. With one exception, these two techniques yielded similar results from all 40 samples tested. In addition, a rare variant form of apo E (phenotype E1) was analyzed separately and incorrectly diagnosed as E2 by the Hha I digestion method; the anticipated mutation in the codon 127 was, however, confirmed by demonstration of a new Taq I restriction site in this variant gene. These data confirm that the common isoforms of apo E usually arise from genetic variation of the codons 112 and 158 and demonstrate the feasibility of the PCR technique in apo E genotyping.
Determination of reference values for serotonin concentration in platelets of healthy newborns, children, adults, and elderly subjects by HPLC with electrochemical detection
Authors: Flachaire E , Beney C , Berthier A , Salandre J , Quincy C , Renaud B 
Published: January 2020

Abstract
We adapted a high-performance liquid chromatographic method with electrochemical detection (Clin Chim Acta 1984;139:1-12) to the determination of platelet serotonin. We used this method to determine platelet serotonin reference values in a healthy population, measuring platelet serotonin concentration in the following subjects: 31 newborns (16 girls, 15 boys); 41 children (11 girls, 30 boys), ages 20 months to 15 years; 56 adults (26 women, 30 men), ages 20 to 58 years; and 20 elderly subjects (16 women, four men), ages 65 to 94 years. There was no significant difference in platelet serotonin concentration between sexes in each age group. However, significant changes (P less than 0.001) were observed between the newborns (mean +/- SD: 1.67 +/- 0.74 nmol/10(9) platelets) and the children (4.09 +/- 1.04) or the adults (3.81 +/- 0.87). Moreover, the platelet serotonin concentration in the elderly subjects (2.57 +/- 1.12) was significantly (P less than 0.001) lower than in the adults and children and significantly higher (P less than 0.01) than in the newborns. Such age-related differences must be taken into consideration when data from neurological or psychiatric patients and control subjects are compared.
Relationship between delta checks for selected chemistry tests
Authors: Lacher D A 
Published: January 2020

Abstract
The correlation between delta differences for 20 serum chemistry tests was calculated for 2400 samples from 288 patients. There were 12 pairs of chemistry tests for which correlation coefficients of the delta checks exceeded 0.25; aspartate aminotransferase and alanine aminotransferase had the highest Pearson correlation coefficient, 0.915. The highest negative, indirect, correlation was between the delta checks of bicarbonate and chloride (-0.219). The relationship between delta differences may be used as a quality-control technique to detect analytical errors.
A monoclonal-antibody-based enzyme-linked immunosorbent assay of lipoprotein(a)
Authors: Wong W L , Eaton D L , Berloui A , Fendly B , Hass P E 
Published: January 2020

Abstract
Lipoprotein(a) [Lp(a)] is a low-density lipoprotein (LDL)-like lipoprotein particle recently described as a risk factor for premature coronary heart disease, stroke, and atherosclerosis. Structurally, Lp(a) is similar to LDL in that it has comparable lipid composition and contains apolipoprotein B-100 (apo B-100). In addition, Lp(a) contains the glycoprotein apolipoprotein(a) [apo(a)], which is disulfide-linked to apo B-100. The recent awareness of a striking correlation between atherosclerosis and concentrations of Lp(a) in plasma prompted our development of an accurate quantitative assay for plasma Lp(a), a monoclonal-antibody-based enzyme-linked immunosorbent assay for Lp(a) that is shown to be sensitive, precise, and highly specific. The response to several isoforms of Lp(a) is linear, and as many as 80 samples can be quantified on one plate. This easily performed assay is suitable for use in the clinical laboratory and for screening large populations.
Activity of phospholipase A2 in plasma increases in uremia
Authors: Costello J , Franson R C , Landwehr K , Landwehr D M 
Published: January 2020

Abstract
We measured phospholipase A2 (PLA2; EC 3.1.1.4) activity in normal and uremic plasma, using [1-14C]oleate-labeled autoclaved Escherichia coli as substrate. Hydrolysis of bacterial phospholipid by crude plasma from both groups was optimal at pH 5.5, was specific for the 2-acyl position of phospholipids, and had an absolute requirement for calcium. Activity was greatest in the presence of added Ca2+, 5 mmol/L, but this increase was inhibited by several divalent cations (Mg2+, Zn2+, Cu2+, Ba2+, Co2+, Pb2+, Fe2+) and by Fe3+. PLA2 activity was also inhibited by heparin at acid and alkaline pH, normal plasma being more sensitive than uremic plasma to this inhibition. Enzyme activity in undiluted plasma was eightfold greater in uremic than in normal plasma. Dilution of plasma by two to fourfold increased the total activity of both normal and uremic plasma. However, the relative differences in total activity between the groups remained constant (eight- to 11-fold). The cause and consequences of the increased PLA2 activity in uremia remain to be established.
A unitized enzyme-labeled immunometric digoxin assay suitable for rapid testing
Authors: Sommer R G , Belchak T L , Bloczynski M L , Boguslawski S J , Clay D L , Corey P F , Foltz M M , Fredrickson R A , Halmo B L , Johnson R D 
Published: January 2020

Abstract
An enzyme-labeled immunometric assay has been developed for measuring digoxin concentrations in serum or plasma. Unitized, compartmentalized reagents are used with an automated sample-processing instrument. The enzyme activity of the processed sample, which is directly proportional to the digoxin concentration, is measured by using a reagent strip and the Ames Seralyzer reflectance photometer. The test takes less than 15 min, and digoxin concentrations are calculated from a two-point calibration line stored in the instrument. Within-run CVs for controls at four concentrations ranged from 2.3% to 3.8%; between-run CVs were from 1.5% to 2.6%. Results obtained with clinical serum samples correlated well (r greater than 0.96) with those obtained by fluorescent polarization immunoassay (Abbott TDx) and RIA (Clinical Assays and NML). This rapid and convenient method for monitoring digoxin concentrations in serum or plasma is particularly well suited for decentralized sites such as emergency rooms, urgent-care centers, and physicians' offices.
Automated measurement of two indicators of human selenium status, and applicability to population studies
Authors: McMaster D , Bell N , Anderson P , Love A H 
Published: January 2020

Abstract
We developed an automated system consisting of a centrifugal analyzer and an atomic absorption spectrophotometer with Zeeman background correction to produce a profile of the concentrations of selenium and the selenium-dependent enzyme glutathione peroxidase (EC 1.11.1.9) in serum and whole blood, for use in epidemiological surveys in Northern Ireland. No pretreatment of samples other than dilution is required, and at least 35 subjects can be screened within 24 h of venesection. For selenium in serum the between-run CV was 5.7% and 4.4% within run. For selenium in whole blood the CV was 5.9% between runs, 5.7% within run. The within-run CV for glutathione peroxidase was 0.92% for serum, 1.18% for blood; the between-run CV was 8.3% for serum. To illustrate the use of the system, we present the profile of selenium and glutathione peroxidase in serum and whole blood from 100 adults 25 to 64 years old.
Establishment of an external quality-assessment scheme for amino acid analyses: results from assays of samples distributed during two years
Authors: Rattenbury J M , Townsend J C 
Published: January 2020

Abstract
Ten different samples of lyophilized plasma and two of liquid urine were distributed during two years to 26 laboratories performing quantitative amino acid analyses in a scheme designed to provide external quality assessment. After each distribution, statistical summaries and performance scores based on delta standard deviations and percentage biases from the all-laboratory trimmed means were returned to participants, who also received annual performance summaries based on their accumulated results. Coefficients of variation calculated from returns across all the samples ranged from 13% for glycine to 65% for methionine. Automated ion-exchange amino acid analyzers with ninhydrin detection appeared to perform better than other methods, although there was no clearly superior method and no model of analyzer clearly outperformed the others. These exercises demonstrate that there is room for improvement in the performance of quantitative amino acid analyses and that individual expertise may be more important in maintaining good performance than the choice of method or analyzer.
Immunosuppressive activity of cyclosporine metabolites compared and characterized by mass spectroscopy and nuclear magnetic resonance
Authors: Copeland K R , Yatscoff R W , McKenna R M 
Published: January 2020

Abstract
Eight cyclosporine (CsA) metabolites were isolated from the urine of renal-transplant patients by high-pressure liquid chromatography. Structure and purity of the metabolites were assessed by fast atomic bombardment/mass spectroscopy, by proton nuclear magnetic resonance (NMR), and, when the quantity of metabolites permitted, by 13C-NMR. The immunosuppressive activities (I) of the metabolites were tested in three separate in vitro systems: primary and secondary mixed lymphocyte reactions as well as by a mitogen-stimulated system. The I, as measured by comparing the concentration of each metabolite required for 50% inhibition of incorporation of [3H] thymidine, varied among the assay systems, as did the ranking of I among the test systems. In general, the I of most metabolites in all assay systems were less than 10% of that for CsA. Metabolites with single modifications exhibited the greatest I; e.g., that of M-17 was congruent to 16% of that of CsA (potency ratio 0.16) in a secondary mixed lymphocyte reaction. The significance of these findings in relation to therapeutic monitoring of CsA is discussed.
Selection of medically useful quality-control procedures for individual tests done in a multitest analytical system
Authors: Koch D D , Oryall J J , Quam E F , Feldbruegge D H , Dowd D E , Barry P L , Westgard J O 
Published: January 2020

Abstract
Quality-control (QC) procedures (i.e., decision rules used, numbers of control measurements collected per run) have been selected for individual tests of a multitest analyzer, to see that clinical or "medical usefulness" requirements for quality are met. The approach for designing appropriate QC procedures includes the following steps: (a) defining requirements for quality in the form of the "total allowable analytical error" for each test, (b) determining the imprecision of each measurement procedure, (c) calculating the medically important systematic and random errors for each test, and (d) assessing the probabilities for error detection and false rejection for candidate control procedures. In applying this approach to the Hitachi 737 analyzer, a design objective of 90% (or greater) detection of systematic errors was met for most tests (sodium, potassium, glucose, urea nitrogen, creatinine, phosphorus, uric acid, cholesterol, total protein, total bilirubin, gamma-glutamyltransferase, alkaline phosphatase, aspartate aminotransferase, lactate dehydrogenase) by use of 3.5s control limits with two control measurements per run (N). For the remaining tests (albumin, chloride, total CO2, calcium), requirements for QC procedures were more stringent, and 2.5s limits (with N = 2) were selected.
Liver- and bone-derived isoenzymes of alkaline phosphatase in serum as determined by high-performance affinity chromatography
Authors: Anderson D J , Branum E L , O'Brien J F 
Published: January 2020

Abstract
To separate liver and bone alkaline phosphatase (ALP) isoenzymes in human serum, we used high-performance affinity chromatography (HPAC) on a column of wheat-germ lectin conjugated to 7-microns-diameter silica particles and an eluent containing N-acetyl-D-glucosamine (NAG). On-line spectrophotometric detection of ALP involved pumping diethanolamine-buffered p-nitrophenyl phosphate solution post-column. Bone and liver isoenzymes could be separated into two peaks with only 10% overlap when an exponential gradient was used. A linear-step gradient separated 80.9% of liver ALP and 91.6% of bone ALP in two distinct peaks. True bone and liver ALP peak areas for the linear-step gradient were determined by using correction factors, because each peak contained a co-eluted portion of the other ALP isoenzyme. The detection limit improved 10-fold over those of other techniques for ALP isoenzymes, owing to the relatively large sample that could be applied to the column. Correlation with a urea-inactivation procedure was reasonable for patients' serum samples (r = 0.98 and 0.79 for liver ALP and bone ALP, respectively).
Screening trace elements and electrolytes in serum by inductively-coupled plasma emission spectrometry
Authors: Melton L A , Tracy M L , Möller G 
Published: January 2020

Abstract
This rapid, accurate procedure for trace elements and electrolytes in serum requires little sample preparation: to 1 mL of serum a single reagent is added that contains trichloroacetic and hydrochloric acids for protein precipitation, hydroxylamine sulfate for iron reduction, and yttrium as the internal standard. After centrifugation, the supernates are directly analyzed for Na, K, Mg, Ca, Pi, Fe, Cu, and Zn by inductively-coupled plasma emission spectrometry. The CVs were respectively 7.9%, 8.4%, 8.6%, 10.0%, 9.0%, 9.4%, 9.0%, and 9.0% for five assays of National Institute of Standards and Technology Standard Reference Material (SRM) no. 1598, Bovine Serum. Analytical recoveries ranged from 92% to 107% for both SRM 1598 and commercial control serum.
Cholesterol screening: comparative evaluation of on-site and laboratory-based measurements
Authors: Bachorik P S , Rock R , Cloey T , Treciak E , Becker D , Sigmund W 
Published: January 2020

Abstract
We measured cholesterol in capillary blood samples from 9683 volunteers over a four-day on-site community screening program, using the "Reflotron" desk-top analyzer (Boehringer-Mannheim Diagnostics, Indianapolis, IN). We also measured cholesterol in venous blood samples from 3% of those screened (a) with the Reflotron at the screening sites, (b) in a qualified hospital clinical laboratory, and (c) in a Centers for Disease Control standardized lipoprotein research laboratory. The sensitivity (and specificity) of the Reflotron measurements, with use of the lipoprotein laboratory measurements as the point of reference, was 0.95 (0.73) in capillary blood samples and 0.88 (0.93) in venous blood samples, compared with 0.99 (0.87) in the hospital clinical laboratory. The Reflotron measurements correlated less well with the lipoprotein laboratory values in both venous blood (r = 0.91) and capillary blood (r = 0.89) samples than did the clinical laboratory values (r greater than 0.99). Furthermore, the capillary blood measurements averaged 7% higher than venous measurements when both kinds of samples were analyzed in the Reflotron.
Rapid purification of human DNA from whole blood for potential application in clinical chemistry laboratories
Authors: Adeli K , Ogbonna G 
Published: January 2020

Abstract
A simple, rapid method for isolating human DNA has been developed, which can be routinely used in clinical chemistry laboratories. The entire procedure takes less than 90 min, and as many as 12 blood samples can be handled in one cycle. One milliliter of EDTA-treated blood is lysed and centrifuged to yield a nuclear fraction. The nuclear pellet is treated with sodium dodecyl sulfate/urea and phenol/chloroform to remove contaminating proteins, then the crude DNA extract is purified by use of a Sephadex G-25 spin-column. Typical 260 nm/280 nm absorbance ratio (used to assess purity) and yield for DNA so purified were 1.84 and 24.5 micrograms/mL, respectively. Within- and between-day CVs for recovery of DNA from pooled blood were 8% and 11% respectively. Such DNA preparations were found quite suitable for digestion by a variety of restriction endonucleases and for restriction fragment length polymorphism analysis. We are using this method to isolate DNA from whole blood of myocardial infarction patients for studies on the apolipoprotein B gene.
Immunoradiometric assay of parathyrin with polyclonal and monoclonal region-specific antibodies
Authors: Bouillon R , Coopmans W , Degroote D E , Radoux D , Eliard P H 
Published: January 2020

Abstract
In this immunoradiometric assay (IRMA) of parathyrin (PTH) a polyclonal anti-amino-PTH(1-34) is the capture antibody and a radiolabeled monoclonal anti-hPTH(44-68) is the second antibody. Gel filtration of serum from a hyperparathyroid patient yielded only a single peak of PTH, corresponding to the elution position of synthetic PTH(1-84). Healthy elderly individuals (ages 78 +/- 5 y, mean +/- SD, n = 45) had PTH concentrations (21 +/- 13 ng/L) not significantly higher than those from healthy younger (38 +/- 11 y) adults (20 +/- 8 ng/L, n = 94). Assay results agreed well with those obtained with a carboxyl-terminal PTH assay both in normal subjects (r = 0.63, P less than 0.001) and in patients with primary hyperparathyroidism (r = 0.59, P less than 0.001). Both assays equally discriminated patients with surgically confirmed primary hyperparathyroidism from normal individuals, but the PTH(1-84) IRMA also allowed a nearly absolute discrimination between normal subjects and patients with primary hypoparathyroidism (undetectable serum PTH in 18 of 21 cases) and secondary hypoparathyroidism (caused by hypercalcemia that was caused by a malignant tumor, PTH 1.3 +/- 1.3 ng/L, n = 32). Moreover, the PTH(1-84) IRMA is more sensitive (detection limit in serum, 0.8 ng/L) and easier and quicker to perform than the carboxyl-terminal assay.
Lipoprotein lipids in women with androgen excess: independent associations with increased insulin and androgen
Authors: Wild R A , Applebaum-Bowden D , Demers L M , Bartholomew M , Landis J R , Hazzard W R , Santen R J 
Published: January 2020

Abstract
Concentrations of triglycerides are increased and concentrations of high-density lipoprotein (HDL) cholesterol are low in women with hyperandrogenism. These alterations could be related to excessive androgen or estrogen, to hyperinsulinism, or to a combination of these abnormalities. We examined their independent influences on lipids in 21 women with hyperandrogenism, subgrouped according to apparent source of androgen excess. Results for lipid, androgen, and insulin did not differ among subgroups, so these data were pooled. Free plus albumin-bound testosterone (uT) was correlated with triglycerides (r = 0.69, P less than 0.01) and HDL cholesterol (r = -0.56, P less than 0.01). Both triglycerides (r = 0.66, P less than 0.01) and HDL cholesterol (r = -0.48, P less than 0.05) were also correlated with insulin measured during fasting. Partial correlation revealed that, after adjusting for insulin, lipids were associated with uT. This suggests that androgen excess is independently related to lipid excess. Insulin also was correlated with lipids when adjusted for uT. Free plus albumin-bound estradiol was not associated with any of the lipids. We conclude that altered lipids in women with hyperandrogenism result from the independent effects of androgen and insulin.