bluuebunny/crossref_metadata_2025_split huggingface dataset:
Sensitive time-resolved fluorescence immunoassay of somatotropin in serum
Authors: Kahan I , Papanastasiou-Diamandi A , Ellis G , Makela S K , McLaurin J , D'Costa M , Diamandis E P 
Published: January 2020

Abstract
We describe a new "sandwich"-type non-isotopic immunoassay for human somatotropin (GH, growth hormone) in serum. In the assay, GH is captured by a monoclonal antibody immobilized in a white microtiter well and simultaneously reacted with a second biotinylated monoclonal antibody. The degree of binding of biotinylated antibody, which increases with increasing amount of GH in the sample, is quantified by adding streptavidin labeled with the europium chelate of 4.7 - bis(chlorosulfophenyl) - 1.10 - phenanthroline - 2.9 - dicarboxylic acid. The fluorescent complex on the solid phase is then measured by excitation at 337.1 nm (nitrogen laser) and monitoring the emission at 615 nm in a gated fluorometer/analyzer. The proposed procedure has short incubation times (less than 4 h protocol), uses only 25 microL of serum per microtiter well, and gives precise and accurate results. The method was clinically evaluated with samples obtained from pediatric patients undergoing investigation for growth abnormalities and from a patient with acromegaly.
Sample evaporation and its impact on the operating performance of an automated selective-access analytical system
Authors: Burtis C A 
Published: January 2020

Abstract
In a series of gravimetric and analytical experiments I measured how much analytical error is ascribable to sample evaporation during routine operation of the Dimension analytical system. This error, a time-dependent function, is related to the volume of liquid placed in either a 0.5- or 2.0-mL cup and whether the cup is capped. For the 0.5-mL cups, analytical error at a 4-h residence time ranged from 12% to 16% (uncovered) to 8% to 13% (covered), whereas errors for the 2.0-mL cup ranged from 6% to 10% (uncovered) to 2% to 5% (covered). To minimize evaporative loss, I recommend (a) protecting the surface areas of the samples, (b) using larger samples, and (c) minimizing the time that a sample is in its cup before and during analysis.
Alpha 1-acid glycoprotein decreases recovery of total protein in urine when trichloroacetic acid is used to precipitate the proteins
Authors: Beilby J P , O'Leary B A 
Published: January 2020

Abstract
Total urine protein was measured in 132 samples by an automated benzethonium chloride method and the Ponceau-S/trichloroacetic acid (PS/TCA) method. Of these, 27% gave a result 0.1 g/L or more higher by the benzethonium chloride method. Of this 27%, most contained an abnormally high concentration of the acute-phase reactant, alpha 1-acid glycoprotein. By assaying urine containing added alpha 1-acid glycoprotein and albumin, we found that alpha 1-acid glycoprotein causes the PS/TCA method to underestimate the total urine protein concentration, whereas the benzethonium chloride method is unaffected. Not all urinary albumin was precipitated by TCA when alpha 1-acid glycoprotein was present. Therefore, protein methods in which trichloroacetic acid is used as a concentrating step before the assay will underestimate total urine protein when the concentration of alpha 1-acid glycoprotein is high.
Blood cholesterol concentration: fingerstick plasma vs venous serum sampling
Authors: Greenland P , Bowley N L , Meiklejohn B , Doane K L , Sparks C E 
Published: January 2020

Abstract
To assess whether venous and fingerstick blood samples yield similar cholesterol concentrations, we obtained both types of samples simultaneously in 108 volunteers participating in a cholesterol screening program. All samples were analyzed by the same enzymatic method in a standardized laboratory, and pairs of simultaneous samples were measured in the same laboratory run. Cholesterol concentrations in fingerstick-derived plasma were consistently higher than in the venous serum (P less than 0.0001), by a positive bias averaging 3.6%. Cholesterol values in fingerstick plasma also were higher than cholesterol results for venous serum placed in a capillary collection tube (average bias +2.4%). The positive bias of fingerstick plasma vs venous serum results appears to be at least partly due to specimen handling, although a true physiological difference between venous and fingerstick cholesterol concentrations is probably also involved. If a positive bias of this magnitude from fingerstick blood sampling is left unadjusted, substantial numbers of people will be labeled "at risk" and referred to physicians when their true values were actually within the acceptable range.
Urinary hydroxyproline in infants with and without fractures/rickets
Authors: Koo W W , Krug Wispe S K , Succop P , Champlin A , Sherman R , Berry H 
Published: January 2020

Abstract
Molar ratios of peptide-bound and free hydroxyproline:creatinine (OHPr:Cr) in urine were measured at 3, 6, 9, and 12 months of age in two groups of very-low-birthweight (VLBW less than or equal to 1500 g) infants. Group A (15 infants) had radiographically confirmed fractures and (or) rickets (F/R); Group B (17 infants) did not. The urinary peptide-bound OHPr:Cr ratio varied widely within groups and was greatest at three months in both groups: A = 0.81 +/- 0.45 and B = 0.55 +/- 0.32 (mean +/- SD). The ratio decreased with increasing postnatal age for each group but was not statistically different between groups throughout the study. The urinary free OHPr:Cr ratio also was greatest at age three months (A = 0.32 +/- 0.15 and B = 0.53 +/- 0.46), rapidly decreasing afterwards, and was not statistically different between groups throughout the study. We conclude that, in VLBW infants, bone turnover as indicated by the urinary peptide-bound OHPr:Cr ratio is highest during early infancy; however, the wide range of values for this ratio suggests that its use alone is not sufficient for detection of F/R in VLBW infants. The rapid decrease in free OHPr:Cr ratio is presumably related to the maturation of renal tubular function.
Competitive heterogeneous enzyme immunoassay for theophylline by flow-injection analysis with electrochemical detection of p-aminophenol
Authors: Gil E P , Tang H T , Halsall H B , Heineman W R , Misiego A S 
Published: January 2020

Abstract
A competitive enzyme-linked immunoabsorbent assay based on the flow-injection amperometric detection of p-aminophenol has been investigated with use of the materials and general procedure of a commercial kit for the determination of theophylline in human serum. The antibody is immobilized on glass beads, and the enzyme label is alkaline phosphatase (EC 3.1.3.1). The high currents generated during the electrochemical detection allowed a rapid (35 min) and simple determination of theophylline throughout its therapeutic range (10-20 mg/L) and also in the subtherapeutic range (detection limit of about 80 micrograms/L).
Dichlorobenzoquinone chloroimine colorimetry of uric acid in urine
Authors: Miller P , Oberholzer V 
Published: January 2020

Abstract
In this nonenzymatic colorimetric method, dichlorobenzoquinone chloroimine is used as a color reagent to measure uric acid in urine. This method is less subject to the interferences that make enzymatic methods unreliable and is inexpensive, simple, and fast.
Retention of antimony in skin biopsies of leishmaniasis patients after treatment with N-methylglucamine antimoniate
Authors: Dorea J G , Merchan-Hamann E , Ryan D E , Holzbecher J 
Published: January 2020

Abstract
Leishmaniasis patients were treated with N-methylglucamine antimoniate by intravenous injections of 10 or 20 mg of Sb per kilogram of body weight per day for 10 or 20 days. Digests of skin biopsies taken from the site of lesion before and after treatment were analyzed for antimony by instrumental neutron activation (INAA). The detection limit of the assay was 20 ng, and no Sb could be measured in digests of samples (less than 2.45 mg) taken before treatment. Biopsies taken after injections of Sb showed concentrations in the range of 8.32 to 70.68 ng/g skin. We discuss the usefulness of INAA in the study of Sb in small samples of tissues.
Measurement of pepsinogen group I in endoscopic gastroduodenal biopsies
Authors: Plebani M , Di Mario F , Dal Santo P L , Faggian D , Germana B , Vianello F , Naccarato R , Burlina A 
Published: January 2020

Abstract
A technique for determining pepsinogen group I (PGI) concentrations in endoscopic gastroduodenal biopsies is described and validated. This method has a good precision (intra-assay CVs 4.1% to 8.4%; between-assay CVs 4.6% to 9.3%), and analytical recovery is satisfactory (94% to 102%). Results were uninfluenced by the storage interval of the samples. At PGI-producing sites (i.e., fundus, corpus) peptic activity was greater than in the antrum and duodenum. PGI concentrations in serum and in gastroduodenal biopsies were not correlated. This simple, reliable method can quantify better than other assays peptic activity in humans, without being influenced by gastric secretory volumes. It will also facilitate prospective studies on the effect of various secretagogues in vivo and evaluations of the influence of anti-ulcer drugs on peptic secretion.
Effect of isoenzyme composition on Kodak Ektachem test results for creatine kinase, lactate dehydrogenase, alkaline phosphatase, and amylase
Authors: Gorus F K , De Pree L 
Published: January 2020

Abstract
Seventy-four patients' sera containing macroenzymes or other uncommon enzyme forms were analyzed for total creatine kinase, lactate dehydrogenase, alkaline phosphatase, or amylase activity on an Ektachem 700XR analyzer. The Ektachem test results correlated well with activities obtained by means of liquid reagents on RA-1000. For each enzyme tested, linear-regression analysis yielded data comparable with those for sera containing exclusively the more common isoenzymes. Increased concentrations of intestinal ALP, however, resulted in higher activities of total ALP being reported for the Ektachem.
Determination of urinary norepinephrine and epinephrine by liquid chromatography with fluorescence detection and pre-column derivatization
Authors: Moleman P , van Dijk J 
Published: January 2020

Abstract
We assayed norepinephrine and epinephrine by utilizing solvent extraction, fluorescence derivatization, and "high-performance" liquid chromatography. A 100-microL aliquot of urine was extracted twice according to Smedes et al. (J Chromatogr 1982;231:25-39) and subsequently incubated with 1,2-diphenylethylenediamine. A 100-microL aliquot of the resulting mixture was injected into a reversed-phase column, and norepinephrine and epinephrine were detected fluorometrically. The within-day CV was 3.0-4.0% and the between-day CV 3.4-7.1% for normal and high concentrations of both analytes. At low concentrations (15-40 nmol/L) these CVs were 4.2-6.8% and 6.8-9.2%, respectively. The detection limit of the assay was less than 0.4 nmol/L for each analyte. We discuss the critical steps for extraction, derivatization, and chromatography. The present method combines the qualities of high precision, specificity, and sensitivity.
Effects of methylamine and heparin on a rapid chromogenic assay of C1-esterase inhibitor in plasma
Authors: Munkvad S , Jespersen J , Gram J , Overgaard K , Rånby M 
Published: January 2020

Abstract
We describe a rapid assay of C1-esterase inhibitor (C1-inh) activity in plasma. After adding purified C1s serine protease (EC 3.4.21.42) in excess to plasma, we determine the residual C1s activity towards a new chromogenic tripeptide, CH3CO-Lys(CbO)-Gly-Arg-pNA. Optimal conditions include the addition of methylamine (final concentration 0.12 mol/L) to reduce the potential inhibitory capacity of alpha 2-macroglobulin towards C1s and the addition of heparin (final concentration 3000 int. units/L) to enhance the reaction of C1s with C1-inh. The correlation with C1-inh antigen concentrations in plasma was excellent. The estimated interassay CV was 4.3%, whereas the intra-assay CV was 2.0% for activity concentrations within the range of normal individuals (means +/- 2 SD: 70-124%), 1.3% at lower concentrations. The method is more convenient, rapid, and precise than previous methods, and C1-inh activity in plasma can be assessed within 30 min. We found that concentrations of C1-inh in plasma were low during open-heart bypass surgery.
Determination of reference intervals for serum magnesium
Authors: Duncanson G O , Worth H G 
Published: January 2020

Abstract
Using a discrete analyzer and a dye-binding method, we measured magnesium in 800 patients' samples received for routine analysis. By excluding data from samples for which the calcium and (or) alkaline phosphatase values were outside defined reference limits, we established a reference interval for magnesium. Because the data showed a gaussian distribution, we could use parametric analysis to establish age-related intervals for both males and females.
High predictivity of galactosyl-hydroxylysine in urine as an indicator of bone metastases from breast cancer
Authors: Moro L , Gazzarrini C , Modricky C , Rovis L , de Bernard B , Galligioni E , Crivellari D , Morassut S , Monfardini S 
Published: January 2020

Abstract
We measured the urinary excretion of galactosyl-hydroxylysine (GH) and hydroxyproline in two groups of women with breast cancer, with (M+, n = 24) and without (Mo, n = 30) clinical, scintigraphic, or radiological evidence of bone metastases. Both these compounds are excreted in larger amounts in the M+ group than in the Mo patients. However, GH, which is a specific marker for bone collagen, provides better predictivity for bone metastases than does hydroxyproline: 92% sensitivity and 90% specificity vs 74% and 79%, respectively, for hydroxyproline.
Screening with zinc protoporphyrin for iron deficiency in non-anemic female blood donors
Authors: Jensen B M , Sandø S H , Grandjean P , Wiggers P , Dalhøj J 
Published: January 2020

Abstract
Iron-depleted donors are at increased risk of developing anemia; if these donors could be identified by a screening test, iron supplementation or decreased donation frequency could be considered. Tests to determine serum ferritin, blood hemoglobin, and erythrocyte (Erc)-zinc protoporphyrin concentrations were examined in 679 consecutive female blood donors to identify donors with non-anemic iron deficiency. The test to determine serum ferritin is expensive and slow, whereas the two latter tests are rapid and less costly and could therefore be used for screening. Women in the fertile age groups had the lowest average serum ferritin values. In all, 93 women (13.7%) had depleted iron stores, as indicated by serum ferritin concentrations less than 14 micrograms/L. In these women, a much better correlation was found between Erc-zinc protoporphyrin and serum ferritin (rs = -0.49, P less than 0.001) than between blood hemoglobin and serum ferritin (rs = 0.31, P less than 0.01). These findings suggest that measurement of Erc-zinc protoporphyrin is superior to that of blood hemoglobin in identifying donors with non-anemic iron deficiency.
Radioligand binding assay of epidermal growth factor receptor: causes of variability and standardization of the assay
Authors: Dittadi R , Gion M , Brazzale A , Bruscagnin G 
Published: January 2020

Abstract
Although experimental evidence indicates a probable role of epidermal growth factor receptor (EGFr) in clinical oncology, no standardized method for its determination has been yet described, and discrepant results have been reported in clinical studies. In standardizing a radioligand binding assay for EGFr, we evaluated the causes of variability in each step of the assay. Entrapment of EGFr in the nuclear fraction and contamination of the crude membrane fraction by cytosol protein were eliminated through preliminary purification steps. Both Scatchard and Rosenthal analysis of the saturation reaction of the membrane fraction with a wide range of concentrations of 125I-labeled EGF revealed a double class of binding sites. Study of the saturation reaction showed a partial exchange of 125I-labeled EGF with endogenous EGF within 20 h. The present method--incubation of partly purified membrane fraction with 125I-labeled EGF, 0.5 nmol/L, with and without 100-fold excess of cold EGF, for 20 h at 26 degrees C, followed by centrifugation at 5000 x g for 30 min to separate membrane-bound 125I-labeled EGF--shows good sensitivity, precision, and accuracy; is reasonably simple; and may be suitable for routine clinical use.
Immunoradiometric assay of atrial natriuretic peptide in unextracted plasma
Authors: Tattersall J E , Dawnay A , McLean C , Cattell W R 
Published: January 2020

Abstract
We have developed and validated a two-site liquid-phase immunoradiometric assay (IRMA) of atrial natriuretic peptide (ANP) in unextracted human plasma. Both radiolabeled rabbit anti-ANP IgG and polyclonal mouse anti-ANP must bind to ANP for detection, and the assay is specific for peptides with both an intact C-terminus and a disulfide bridge. The assay sensitivity (detection limit) is 0.96 pmol/L, and the working range is 2.3-300 pmol/L, with the hook effect occurring above 500 pmol/L. Results for diluted plasma from normal subjects and from patients with renal failure paralleled the standard curve; analytical recovery of ANP added to such samples averaged 94%. The between- and within-assay CVs at 8 pmol/L were 10% and 5%, respectively. The assay is sufficiently sensitive and precise to detect the postural change in ANP concentrations in normal subjects.
Age and sex distribution of alkaline phosphatase isoenzymes by agarose electrophoresis
Authors: Van Hoof V O , Hoylaerts M F , Geryl H , Van Mullem M , Lepoutre L G , De Broe M E 
Published: January 2020

Abstract
We separated isoenzymes of alkaline phosphatase (ALP; EC 3.1.3.1) in 1383 sera of normal individuals (ages 4-65 years) by agarose electrophoresis with the Isopal system (Analis). As expected, the predominant isoenzyme in children was of bone origin, and almost all (99%) of the children had low activities of a second bone fraction, "bone variant" ALP. The "bone variant" disappeared after age 17 in girls and after age 20 in boys. The highest (median) bone ALP activity was reached at age 9 to 10 in girls and at age 13 to 14 in boys, followed by a gradual decline in girls and a steep decline in boys. During adulthood, activity of the bone fraction was constant and no significant differences were observed between sexes, neither for bone nor for liver ALP activity. The latter remained almost unchanged throughout life. We observed no high-Mr ALP activity in children, whereas sera from 60% of the adults contained low activities of high-Mr ALP. Intestinal ALP (soluble form) and "intestinal variant" ALP (hydrophobic form) were frequently present, in 21% and 37% of all samples, respectively. No significant differences were observed between age groups and sexes for the intestinal isoenzymes.
Unique structure of glycopeptide from alpha-fetoprotein produced in human hepatoma cell line, as determined by 1H-nuclear magnetic resonance spectroscopy
Authors: Terrana B , Tecce M F , Manetti R , Ceccarini C , Lamba D , Segre A L 
Published: January 2020

Abstract
Determination of alpha-fetoprotein is used in diagnosis of tumors and neural tube defects. A good reliable source of alpha-fetoprotein would be an obvious advantage to the preparation of diagnostic reagents and their standardization. We have recently developed a method for the production of alpha-fetoprotein from a human hepatoma cell line. This method, which is suitable for scaling up, allowed us to produce 40 g of alpha-fetoprotein from culture supernatant liquid through a simple purification procedure. We have previously shown this protein to be identical to alpha-fetoprotein produced from other sources. However, because the presence of different glycoforms has been reported in alpha-fetoprotein preparations, both from human sources and from other species, it was important to establish the type and extent of glycosylation of alpha-fetoprotein prepared by our method. By using 1H-NMR spectroscopy we were able to establish that our product contains a single N-linked biantennary, fully sialylated complex-type oligosaccharide, typical of human hepatomas.
Diagnosis of perioperative myocardial infarction by considering relationship of postoperative electrocardiogram changes and enzyme increases after coronary bypass operation
Authors: Griesmacher A , Grimm M , Schreiner W , Müller M M 
Published: January 2020

Abstract
We report the results of enzyme determinations in sera from 88 patients, 65 of whom showed inconspicuous reconvalescence, 14 who had myocardial infarction within 24 h (MI 1) after bypass surgery, and nine with myocardial infarction between 24 and 48 h postoperatively (MI 2). We wanted to determine whether the consequent measurement of activities of total creatine kinase (CK), CK isoenzyme MB (CK-MB), lactate dehydrogenase, alpha-hydroxybutyrate dehydrogenase, and aspartate aminotransferase, conducted as a part of routine laboratory diagnostics, provided meaningful information for diagnosing infarcts besides that obtained from the electrocardiogram. The postoperative mean values of the enzyme activities in blood were significantly different among the three groups; however, only a combined evaluation of CK and CK-MB by means of a discriminant analysis allowed the prediction of MI (sensitivity: MI 1 = 98.5%, MI 2 = 95.4%; specificity: MI 1 = 71.4%, MI 2 = 81.8%). CK greater than 600 U/L or CK-MB greater than 45 U/L supports the diagnosis of acute MI.
Use of polymerase chain reaction to detect heterozygous familial hypercholesterolemia
Authors: Keinänen M , Ojala J P , Helve E , Aalto-Setälä K , Kontula K , Kovanen P T 
Published: January 2020

Abstract
We used a modification of the polymerase chain reaction (PCR), involving two pairs of oligonucleotide primers, to detect a mutation in the low-density lipoprotein (LDL) receptor gene, commonly occurring among patients with familial hypercholesterolemia (FH) in Finland. This mutation, called FH-Helsinki, involves a large (about 9500 base pairs, bp) deletion in the LDL receptor gene extending from intron 15 to exon 18. For the PCR, one pair of primers was designed to cover both sides of the deletion in its immediate vicinity. In the presence of the deletion, the primers were brought close enough to each other to allow the amplification and electrophoretic detection of a 300-bp amplification product. In the absence of the deletion, no amplification occurred and this band accordingly was not visible in the gel. To render the interpretation of the results unequivocal, we designed a second pair of oligonucleotide primers. This pair of primers allowed another amplification product (158 bp) to appear in samples containing a normal exon 17, i.e., in DNA specimens from healthy subjects and FH heterozygotes with or without the FH-Helsinki deletion. The technique is easy to perform, avoids the use of radioactive reagents, and is applicable to the detection of any extensive DNA deletion.
Chemiluminescence immunoassay of cyclosporine in whole blood
Authors: Stabler T V , Siegel A L 
Published: January 2020

Abstract
We describe a chemiluminescent immunoassay (CLI) for measuring cyclosporine in whole blood. Its sensitivity and accuracy are comparable with those of an RIA method that makes use of the same specific monoclonal antibody. The comparison with the RIA method was excellent: y(RIA) = x(CLI) + 11.24 (r = 0.99). In our procedure the samples are incubated with cyclosporin C-hemisuccinate-aminobutyl-N-ethylisoluminol, antibody, and paramagnetic particles coated with second antibody. After magnetic separation and washing, the samples are incubated with 200 microL of NaOH (2 mol/L) at 60 degrees C for 30 min. The chemiluminescence generated by automated serial injections of solutions of microperoxidase and dilute (2 mL/L) H2O2 is measured for 5 s. The data are processed by using a spline fit of log B/Bo log conversion. This method is easy to perform and avoids the hazards and costs associated with isotopic waste disposal. The label is stable for at least three years.