bluuebunny/crossref_metadata_2025_split huggingface dataset:
Rapid and sensitive chemiluminescent enzyme immunoassay for measuring tumor markers
Authors: Nishizono I , Iida S , Suzuki N , Kawada H , Murakami H , Ashihara Y , Okada M 
Published: January 2020

Abstract
We developed a chemiluminescent enzyme immunoassay (CLEIA) to quantify such tumor markers as carcinoembryonic antigen (CEA), alpha-fetoprotein (AFP), CA19-9, and CA125. We used a novel chemiluminescent substrate, a derivative of 1,2-dioxetane phosphate (AMPPD), to measure alkaline phosphatase as a labeling enzyme to Fab' fragments of antibody. Regardless of the solid phase, i.e., polystyrene beads (6 mm diameter) or ferrite-coated particles (0.3 microns diameter), the standard curves within the dynamic ranges of the conventional RIA or enzyme immunoassay (EIA) were linear in all cases except for those with AFP. Use of the ferrite particles further shortens the immunoreaction, so the assay can be performed in 30 min. In addition, the relationships between concentrations of the marker and chemiluminescent signals for CA19-9, CA125, and CEA were linear up to concentrations about 10-fold greater than the ordinary dynamic ranges. Intra- and interassay CVs (averages for individual analyte) were 2.2%-4.9% and 2.0%-5.8%, respectively. In an analysis of serum samples, results of the CLEIA correlated reasonably well with those of RIA or EIA. The lower limit of detection by CLEIA with ferrite particles was 390 arb. units/L for CA19-9, 990 arb. units/L for CA125, 0.06 micrograms/L for CEA, and 0.03 micrograms/L for AFP. Thus, the sensitivity increased to between two- and 10-fold that of RIA or colorimetric EIA, depending on the analytes.
Development of an enzyme-linked immunosorbent assay of apolipoprotein E-AII complex in plasma
Authors: Tozuka M , Yoshida Y , Tanigami J , Miyachi M , Katsuyama T , Kanai M 
Published: January 2020

Abstract
Measurement of apolipoprotein (apo) E-AII complex in human plasma is important in determining the role of apoE in lipoprotein metabolism. In this paper, we demonstrate a new and simple method to determine apoE-All complex by using an enzyme-linked immunosorbent assay. Anti-apoE IgG (goat) was used as a capture antibody, and captured apoE-All complexes were detected by an anti-apoAll (rabbit) horseradish peroxidase-conjugated anti-rabbit IgG (goat) system. With this method, apoE-All complex was specifically determined without the interference of apoAll and was not affected by apoE monomer less than 250 mg/L. The content of the complex in reference serum, a normolipidemic serum pooled from five subjects with phenotype E3/E3, was arbitrarily defined as 100%. The coefficients of variation were 3.5%-6.3% within assay and 8.8%-11.6% between assays.
High-Performance Liquid-Chromatographic Measurement of Plasma Creatinine in Newborns
Authors: Scott Paul H 
Published: January 2020

Abstract
This HPLC method for measuring plasma creatinine is based on cation-exchange chromatography and is particularly suitable for use with specimens from babies. A short chromatographic run is performed after simple protein precipitation with zinc sulfate and addition of an internal standard, N-methylnicotinamide. The standard curve for the method is linear up to 200 mumol/L, and analytical recovery of added creatinine is between 101% and 103%. Between-batch precision (CV) is less than 3% for mean creatinine values of 103 and 164 mumol/L. The method is free of interference from other metabolic components and drugs commonly used in neonates in routine clinical practice. Using specimens from neonates, I compared this method with a routinely used automated alkaline picrate method (from Randox Labs., performed on a Cobas MIRA analyzer). Linear-regression analysis yielded a correlation coefficient of 0.90, a slope of 1.00, and an intercept of +0.8 mumol/L. This HPLC method for creatinine should be of use in those circumstances where the alkaline picrate method is known to produce dubious results; however, the latter method is probably more suitable for routine use.
Elimination of Matrix Effects in Electrothermal Atomic Absorption Spectrophotometric Determinations of Bismuth in Serum and Urine
Authors: Dean Shaheb-ud , Tscherwonyi Peter J , Riley William J 
Published: January 2020

Abstract
A sensitive and precise electrothermal atomic absorption spectrophotometric method for determining bismuth concentration is described. Protein precipitation and the use of a palladium modifier reduce the problems of foaming and permit the use of a higher ashing temperature. The detection limit of the assay is 0.9 nmol/L. Total CVs (intra- and interassay) for serum ranged from 3.5% to 15.1% and for urine from 4.8% to 14.5% at concentrations of 60.0 and 6.0 nmol/L, respectively. Analytical recoveries of bismuth added to serum and urine were 102% and 103% over the same range. The method is robust and reproducible and can be accurately calibrated with aqueous standards.
Seven Different Assays of Hyaluronan Compared for Clinical Utility
Authors: Lindqvist Ulla , Chichibu Kenji , Delpech Bertrand , Goldberg Ronald L , Knudson Warren , Poole A Robin , Laurent Torvard C 
Published: January 2020

Abstract
To compare six assays of hyaluronan (hyaluronic acid; HYA) in serum, developed in different laboratories, we analyzed 10 samples from each of three groups: healthy persons, patients with primary biliary cirrhosis, and patients with rheumatoid arthritis. All the assays are based on the use of affinity proteins specific for HYA, prepared from cartilage or brain tissue, and are analogous to RIA or enzyme immunoassay techniques. The assay results were of the same magnitude. Although statistical analysis indicated that the methods in some cases deviated significantly from one another, this variation was less than the physiological variation in the healthy population. Therefore, the results of clinical investigations in which the various methods have been used are comparable. The analyses have high specificity and sensitivity for primary biliary cirrhosis but are somewhat less suitable for detecting rheumatoid arthritis. A seventh laboratory, which obtained antibodies to HYA, used these in an RIA to analyze a separate series of serum specimens. Results were in agreement with those obtained by one of the other assays.
Polypeptide Nicks Cause Erroneous Results in Assays of Human Chorionic Gonadotropin Free β-subunit
Authors: Kardana Andrew , Cole Laurence A 
Published: January 2020

Abstract
Pregnancy and trophoblastic disease testing laboratories measure human chorionic gonadotropin (hCG; human choriogonadotropin) free beta-subunit to screen for Down syndrome and to diagnose persistent trophoblastic disease (invasive mole and choriocarcinoma). The results from various laboratories, however, vary widely for similar groups of patients. Average concentrations of free beta-subunit reported for persistent trophoblastic disease, for instance, range from 2.6% to 37% of total hCG. On hCG and its free beta-subunit, peptide bonds can be missing between beta-subunit residues 44 and 45 or between residues 47 and 48 (nicked molecules). To explore the possibility that the disparity in the reported concentrations of free beta-subunit was due to differences in recognition of nicked molecules by different antibodies, we generated 0% and 100% nicked beta-subunit standards and investigated their recognition in three separate immunoassays of free beta-subunit. The immunoassay with antibody 1E5 did not recognize nicked beta-subunit (4% cross-reactivity with nicked beta-subunit) and thus detected only intact beta-subunit. The immunoassay with antibody FBT11 gave similar results with nicked and nonnicked thus standards (96% cross-reactivity with nicked beta-subunit), as did the assay with antibody B204 (73% cross-reactivity with nicked beta-subunit). These two immunoassays thus measured total (nicked + nonnicked) beta-subunit. We used the three immunoassays to examine sera from normal pregnancy, Down syndrome pregnancy, hydatidiform mole, and persistent trophoblastic disease, all of which contain nicked and nonnicked beta-subunit molecules. The results obtained resembled the studies with nicked beta-subunit standards. The results from the FBT11 and B204 assays (total beta-subunit) were highest, results from the 1E5 assay (intact molecules only) being as much as 10 times lower. We conclude that nicks in beta-subunit and the extent of recognition of nicked molecules by different antibodies affect the concentrations reported for free beta-subunit.
Ion-Trap Detection of Volatile Organic Compounds in Alveolar Breath
Authors: Phillips M , Greenberg J 
Published: January 2020

Abstract
We describe a method for the collection and microanalysis of the volatile organic compounds in human breath. A transportable apparatus supplies subjects with purified air and samples their alveolar breath; the volatile organic compounds are captured in an adsorptive trap containing activated carbon and molecular sieve. The sample is thermally desorbed from the trap in an automated microprocessor-controlled device, concentrated by two-stage cryofocusing, and assayed by gas chromatography with ion-trap detection. Compounds are identified by reference to a computer-based library of mass spectra with subtraction of the background components present in the inspired air. We used this device to study 10 normal subjects and determined the relative abundance of the volatile organic compounds in their alveolar breath. The breath-collecting apparatus was convenient to operate and was well tolerated by human volunteers.
Fecal Coproporphyrin Isomers in Hereditary Coproporphyria
Authors: Blake Dennis , McManus Julie , Cronin Virginia , Ratnaike Sujiva 
Published: January 2020

Abstract
To see whether the fecal coproporphyrin III:coproporphyrin I (CIII:CI) ratio (determined by HPLC) would be suitable for screening patients at risk of hereditary coproporphyria (HC), we compared such ratios with the lymphocyte coproporphyrinogen oxidase (EC 1.3.3.3) activities (COOX) in 38 subjects from one large family and two smaller families with HC. The CIII:CI ratio was normal (less than 1.3) in adults with normal COOX (greater than 180 nmol/g of protein per hour) and high (greater than 2) in those with low COOX. Results were difficult to interpret in six of 10 children, who had borderline or low COOX but normal fecal CIII:CI ratios. Five subjects with low COOX and abnormal fecal CIII:CI ratios had normal fecal total porphyrin, indicating that the latter investigation alone is inadequate for family studies. The sample for determining the fecal CIII:CI ratio is easier to obtain and the assay is technically less demanding than COOX. We found the fecal CIII:CI ratio suitable for investigation of adults in a family study, but its usefulness in children needs to be established.
Spectrotype Distributions of Circulating IgG from Patients with Systemic Lupus Erythematosus
Authors: Barbi G , Keil L B , Gaito A D , DeBari V A 
Published: January 2020

Abstract
Pathogenic autoantibodies from patients with systemic lupus erythematosus (SLE) may represent a relatively cationic fraction of IgG. We compared the spectrotype distributions of affinity-purified IgG from the sera of 10 SLE patients and 10 age- and sex-matched control subjects. Purified IgG was subjected to isoelectric focusing between pI 3 and 9. No significant difference was observed for pI 6.0-6.5 and 7.5-8.0. However, control subjects had a higher percent of total IgG at 6.5-7.0 (15.4 +/- 4.0 vs 11.1 +/- 2.0, P = 0.008) and at 7.0-7.5 (22.4 +/- 4.8 vs 18.2 +/- 3.8, P = 0.04) whereas SLE patients had a higher percent of total IgG at 8.0-8.5 (24.3 +/- 3.0 vs 20.5 +/- 4.0, P = 0.03) and at 8.5-9.0 (21.9 +/- 5.9 vs 15.1 +/- 3.7, P = 0.006). Spectrotype distributions of circulating IgG from SLE patients are skewed toward higher pI, providing further evidence of proliferation of B-cell clones that express more cationic IgG in patients with SLE. Longitudinal studies of serum IgG from several patients for &gt; 1 year reveal distinct changes in both cationic and anionic clonotypes, suggesting clonal expansion of antiidiotypes to autoantibodies.
Using Changes in Attributable Risk to Predict Long-Term Efficacy of Simvastatin Treatment
Authors: Vermaak W J , Ubbink J B , Ungerer J P , Marais A D , Firth J , Van Lathem J M , Becker P J 
Published: January 2020

Abstract
It is a common perception that coronary heart disease (CHD) deaths will decline by 2% for every 1% decrease in serum total cholesterol (TC). We investigated the changes in serum TC concentrations obtained with simvastatin, an inhibitor of hydroxymethylglutaryl-CoA reductase, in 70 hypercholesterolemic subjects. Although simvastatin reduced serum TC concentrations by 30% after 6 months, calculations from age- and risk-related TC quintiles show that the absolute difference in attributable CHD deaths would be just &gt; 1%. These results are reminiscent of the experience from several large prospective primary drug trials that demonstrated impressive relative benefit ratios but much smaller absolute differences (&lt; 2%) in CHD endpoints between the treated and untreated groups. This epidemiologically based approach of assessing efficacy and potential long-term benefit of a lipid-lowering drug may be more appropriate and potentially less misleading than just the percentage change of TC (or other lipoproteins) so frequently reported in short-term drug trials.
Transferrin Index: An Alternative Method for Calculating the Iron Saturation of Transferrin
Authors: Beilby J , Olynyk J , Ching S , Prins A , Swanson N , Reed W , Harley H , Garcia-Webb P 
Published: January 2020

Abstract
We surveyed 140 clinical chemistry laboratories in Australia to establish which laboratory methods they used to determine serum iron status: 125 measured serum iron (Fe), 85 measured transferrin (TRF), 47 measured total iron-binding capacity (TIBC), and 14 measured both TRF and TIBC. Of the 55 laboratories routinely reporting TRF saturation (TS), 16 calculated TS directly as (Fe/TIBC) x 100, and 9 used [Fe/(TRF x 2)] x 100. Thirty laboratories measured TRF and converted it to an equivalent TIBC concentration; the derived TIBC was then used to calculate TS. We measured iron, TIBC, and TRF concentrations in 94 control subjects, 59 patients with alcoholic liver disease (ALD), and 20 with proven genetic hemochromatosis (GH). TS was compared with a transferrin index (TI = Fe/TRF) to determine whether both methods were sensitive for GH screening and which method gave the fewest false-positive results with discrimination limits of &gt; 55% and &gt; 1.0, respectively. All GH patients were detected by both TS and TI at these limits. One control subject had a TI &gt; 1.0, whereas three control subjects had a TS &gt; 55%. Nine patients with ALD had a TI &gt; 1.0 and 11 ALD patients had a TS &gt; 55%. Some iron-overload patients had lower than expected TS values compared with TI, possibly because of ferritin interference in the TIBC assay. Also, the precision of the TRF assay was better than that of the TIBC assay: CVs of 1.85-3.68% vs 6.17%. We therefore recommend that calculated TI replace TS in screening for iron overload.
Serotonin Concentrations in Plasma and Variations During the Menstrual Cycle
Authors: Hindberg I , Naesh O 
Published: January 2020

Abstract
We used a specific and sensitive radioenzymatic method to establish a reference interval for the concentration of serotonin in platelet-poor plasma in 98 healthy volunteers (49 men, 49 women). The interval was 0-11 nmol/L with a median of 2.8 nmol/L. No difference in concentration in relation to sex or age was observed. In a group of eight very old volunteers (ages 86-92 years), however, concentrations were increased. In addition, we monitored the plasma concentrations of serotonin in 20 healthy women (ages 26-45 years) through two menstrual cycles. Periovulatory and premenstrual concentrations were greater than the serotonin concentration at the start of menstruation.
Two Novel Nonradioactive Polymerase Chain Reaction-Based Assays of Dried Blood Spots, Genomic DNA, or Whole Cells for Fast, Reliable Detection of Z and S Mutations in the α1-Antitrypsin Gene
Authors: Andresen B S , Knudsen I , Jensen P K , Rasmussen K , Gregersen N 
Published: January 2020

Abstract
Two new nonradioactive polymerase chain reaction (PCR)-based assays for the Z and S mutations in the alpha 1-antitrypsin gene are presented. The assays take advantage of PCR-mediated mutagenesis, creating new diagnostic restriction enzyme sites for unambiguous discrimination between test samples from individuals who are normal, heterozygous, or homozygous for the mutations. We show that the two assays can be performed with purified genomic DNA as well as with boiled blood spots. The new assays were validated by parallel testing with a technique in which PCR is combined with allele-specific oligonucleotide (ASO) probes. In all cases tested the results obtained by the different techniques were in accordance. The new assays can be used for prenatal diagnostics and can be performed directly with boiled tissue samples. Because the new assays are easy to perform and reliable, we conclude that they are well suited for routine diagnosis.
Fluorometric Enzyme Immunoassay of Basic Fibroblast Growth Factor with Monoclonal Antibodies
Authors: Kurobe M , Kato A , Takei Y , Hayashi K 
Published: January 2020

Abstract
We compared three different strategies for measuring basic fibroblast growth factor (bFGF) by fluorometric enzyme immunoassay (EIA). After optimizing conditions, we found that a primary anti-bFGF MAb directly conjugated with peroxidase gave the best detection limit for recombinant bFGF (approximately 30 ng/L, 3 pg/assay tube) in a two-site sandwich assay. The detection limit of methods based on biotinylated primary MAbs or on secondary antibodies followed by streptavidin-conjugated peroxidase was slightly lower than that of the above method. Using the most sensitive EIA examined in this study, we made a preliminary measurement of immunoreactive bFGF in sera of apparently healthy people and found it to be 190 (SD 32) ng/L (n = 48), in agreement with an earlier reported value (30-206 ng/L). Also, the concentration of immunoreactive bFGF in sera was above normal in 19 of 31 patients with stomach cancer.
Abbott IMX Evaluated for Assay of Prostate-Specific Antigen in Serum
Authors: Dnistrian A M , Schwartz M K , Smith C A , Nisselbaum J S , Fair W R 
Published: January 2020

Abstract
We evaluated a new fully automated procedure for quantitative measurement of prostate-specific antigen (PSA) by the Microparticle Enzyme Immunoassay (MEIA) technology developed for the Abbott IMx automated immunoassay system. The performance characteristics of the Abbott IMx PSA assay (y) were evaluated and compared with those of the Hybritech Tandem-E PSA assay (x), a solid-phase two-site immunoenzymometric assay. PSA values for both assays were well correlated (r = 0.99); regression analysis yielded the equation y = 0.92x - 0.23 micrograms/L. The Abbott assay proved reliable and reproducible, as shown by the intra- and interassay coefficients of variation (2.0-3.4% and 3.1-4.7%, respectively). The assay gave a linear standard curve up to 100 micrograms/L and was very sensitive (detected PSA &lt; 0.1 microgram/L). This analytical sensitivity was comparable with that of the Tandem-E PSA assay. Overall, the IMx PSA assay demonstrated the accuracy, precision, linearity, and intermethod correlation required for monitoring patients with prostate cancer.
Washing Erythrocytes to Remove Interferents in Measurements of Zinc Protoporphyrin by Front-Face Hematofluorometry
Authors: Hastka J , Lasserre J J , Schwarzbeck A , Strauch M , Hehlmann R 
Published: January 2020

Abstract
Zinc protoporphyrin (ZPP) is determined by hematofluorometry of whole blood to detect iron deficiency in blood donors. In hospitalized patients, ZPP did not correlate with established markers of iron status. We performed 4500 ZPP measurements with the Aviv front-face hematofluorometer in samples from 475 patients and measured ferritin, transferrin saturation, hemoglobin, and erythrocyte indices. We found that the fluorometric determination is affected by substances dissolved in plasma but that this interference can be eliminated by using washed erythrocytes. In validation tests the within-day variation was &lt; 3.5%; the day-to-day variation was &lt; 6.8%. In 130 healthy persons without iron deficiency, ZPP was &lt; or = 40 mumol/mol heme, which we consider a normal value. Mean ZPP in 46 iron-deficient patients was 256 (SD 105) mumol/mol heme (correlation with ferritin: -0.73; with hemoglobin: -0.85; P &lt; 0.001). When washed erythrocytes are used, the hematofluorometric determination of ZPP is sensitive and specific for detecting iron deficiency in otherwise healthy individuals and hospitalized patients.
Clinical Evaluation of a Pancreatic Lipase Mass Concentration Assay
Authors: van Ingen H E , Sanders G T 
Published: January 2020

Abstract
An immunoactivation assay for determining pancreatic lipase mass concentration was clinically evaluated and compared with results obtained by measuring total amylase and pancreatic amylase activity. A group of 30 patients with pancreatitis was compared with a control group of 32 patients in which this disease was suspected but excluded. Both lipase mass concentration and pancreatic amylase activity exhibit good sensitivity (0.93 each) and specificity (0.94 and 0.97, respectively) at cutoff concentrations of 200 micrograms/L and 200 U/L, respectively. The median increase in lipase mass concentration (37.1 times the upper limit of the reference interval) in the pancreatitis group was higher than that for either total amylase or pancreatic amylase activity (5.94 and 14.5 times, respectively) but showed a similar time to peak value. We conclude that the lipase assay is the method of choice for diagnosing pancreatitis.
Bone-Resorption Markers Galactosyl Hydroxylysine, Pyridinium Crosslinks, and Hydroxyproline Compared
Authors: Bettica P , Moro L , Robins S P , Taylor A K , Talbot J , Singer F R , Baylink D J 
Published: January 2020

Abstract
We compared the clinical performances of four bone-resorption (BR) assays (hydroxyproline, HYP; galactosyl hydroxylysine, GHYL; deoxypyridinoline, DPD; and pyridinoline, PYD) in subjects with different BR rates: normal (adult men and premenopausal women), mildly increased (postmenopausal osteoporotic women), high (Paget disease patients), and very high (children). The discrimination power (Z score) and the accuracy (estimated by receiver-operating characteristic analysis) for GHYL, DPD, and PYD were compared with those for HYP. Discrimination power and accuracy were similar for high- and very-high-BR groups for all four assays. However in the mildly increased-BR group, DPD, GHYL, and PYD showed a higher discrimination power and accuracy than did HYP. The clinical performances of HYP, DPD, GHYL, and PYD are comparable for large changes in BR. For modest changes, DPD, GHYL, and PYD are more accurate and have a higher discrimination power than does HYP.