bluuebunny/crossref_metadata_2025_split huggingface dataset:
Simple and rapid detection of cholesteryl ester transfer protein deficiency by using nonradioisotopic single-strand conformation polymorphism analysis
Authors: Arai T , Tsukada T , Nakayama T 
Published: January 2020

Abstract
We developed a polymerase chain reaction (PCR)-single-strand conformation polymorphism (SSCP) analysis technique by using silver staining to detect the G--&gt;A point mutation of intron 14 of the cholesteryl ester transfer protein (CETP) gene, which is a common mutation in CETP deficiency. After conventional PCR, amplified DNA was heated and subjected to electrophoresis in polyacrylamide gel containing glycerol. The gel was then stained with silver to identify DNA bands. With this technique, we examined 40 subjects with high concentrations of high-density lipoprotein cholesterol (&gt; or = 1 g/L) and found one homozygote and eight heterozygotes. The procedure is suitable for screening because it is quick and does not require radioactive materials or restriction enzymes.
Detergent extraction and enzymatic analysis for fecal long-chain fatty acids, triglycerides, and cholesterol
Authors: Lee M J , Crook T , Noel C , Levinson U M 
Published: January 2020

Abstract
We report a procedure for determining fecal long-chain fatty acids (LCFA), triglycerides, and cholesterol after detergent extraction with 10 mL/L Triton X-100, 6 mL/L Brij 30, and 0.1 mol/L HCl in isotonic saline by enzymatic analysis of the extraction supernates. Mean recoveries of calibrators ranged from 105% to 117%. Assays of fecal extracts were linear with concentration from 0 to 1000 mumol/L for LCFA, from 10 to 90 mumol/L for triglycerides, and from 0 to 300 mumol/L for cholesterol. Within-run CVs were &lt; 3% for medium and high concentrations, and 3%, 14%, and 28% for low concentrations of LCFA, cholesterol, and triglycerides, respectively. Correlation with gravimetric determination resulted in coefficients (r2) of 0.72 for total fat, 0.63 for neutral fat, and 0.66 for nonesterified fat. Assays of extracts and residues prepared for gravimetric determination indicated that approximately 90% of triglycerides, cholesterol, and LCFA were extracted by organic solvents and that the extracts contain a significant amount of other fats. The proposed method appears accurate, precise, specific, and suitable for routine analysis.
Immunological recognition and clinical significance of nicked human chorionic gonadotropin in testicular cancer
Authors: Hoermann R , Berger P , Spoettl G , Gillesberger F , Kardana A , Cole L A , Mann K 
Published: January 2020

Abstract
We studied the physical properties, immunological recognition, and clinical significance of nicked human chorionic gonadotropin (hCGn) in testicular cancer. Upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions, the beta-subunit of hCGn (hCG beta n) dissociated into two peptides, and, by reversed-phase chromatography, hCG beta n was found to be less hydrophobic than hCG beta. Immunologically, hCGn lacked two epitopes specific for holo-hCG (c1, c2), whereas at least five hCG beta epitopes (beta 1-beta 5) were preserved, and, as a result, recognition of hCGn by different hCG assays varied widely. In 309 sera and 88 urine samples from patients with seminomatous or nonseminomatous testicular cancer, hCG-only, hCG+hCGn, and hCG+hCGn+hCG beta+hCG beta n+hCG beta core-fragment assays gave parallel results. hCGn was more abundant in urine than in serum samples. In conclusion, hCGn lacks two conformationally dependent epitopes of hCG, causing a change in hydrophobicity and explaining its failure to react in certain holo-hCG assays. Recognition of hCGn, however, does not seem to be crucial in the routine use of serum hCG as a tumor marker in patients with testicular cancer.
Libyan family with hypercholesterolemia and increased high-density lipoprotein cholesterol in plasma
Authors: Sheriff D S , el Fakhri M , Ghwarsha K 
Published: January 2020

Abstract
Genetic deficiencies of cholesteryl ester transport protein (CETP) and hepatic lipase activities have been associated with hyperalpha-lipoproteinemias. Here we present a family of 11 members, of which 9, including the father, mother, 5 sons, and 2 daughters, show a marked increase in high-density lipoprotein (HDL) cholesterol alone with low plasma concentrations of triglycerides. Analyses of lecithin:cholesterol acyltransferase (LCAT) activity, cholesteryl ester transfer between HDL fractions, hepatic lipase (HL) activity, and lipoprotein lipase (LPL) activity in these cases showed that a decrease in the heparin-releasable HL activity was the possible cause of the marked increase of HDL2 fractions observed in nine of them. Such a defect in HL activity could significantly affect HDL metabolism in particular and lipoprotein metabolism in general. Evidently, a marked increase in serum total cholesterol due to abnormal metabolism of HDL cholesterol, separate from known causes of altered low-density lipoprotein cholesterol metabolism, e.g., a clearance or a receptor defect, is not uncommon. The coordinated action of HL, LCAT, LPL, and CETP may be essential for normal metabolism of plasma lipoproteins.
Ultrasensitive time-resolved immunofluorometric assay of glutathione transferase alpha in serum
Authors: Tiainen P , Karhi K K 
Published: January 2020

Abstract
Glutathione transferase Alpha (GSTA) is a very sensitive marker of acute centrilobular liver damage. We purified glutathione transferases from human liver and separated the isoenzymes. Polyclonal rabbit antibodies specific for Alpha-class isoenzymes were produced and labeled with Eu(3+)-chelate. We set up a sandwich-type time-resolved immunofluorometric assay (TR-IFMA) to measure GSTA in serum. The detection limit was 0.03 microgram/L, and measuring range was from 0.03 to 100 micrograms/L. The reference range for serum GSTA in this assay was 0.7-6.0 micrograms/L for women and 0.7-14 micrograms/L for men. This TR-IFMA is practical, and the modified rapid assay can be completed in 3 h. Therefore it is applicable for diagnostic purposes in acute liver damage, e.g., associated with drug toxicity or hypoperfusion of the liver.
Catecholamines in depression: an update
Authors: Potter W Z , Manji H K 
Published: January 2020

Abstract
Despite extensive research, the biochemical abnormalities underlying the predisposition to and the pathogenesis of affective disorders remain to be clearly established. Efforts to study norepinephrine (NE) output and function have utilized biochemical assays, neuroendocrine challenge strategies, and measures of peripheral blood cell receptors; the cumulative database points to a dysregulation of the noradrenergic system. Depressed patients (in particular, melancholic, unipolar subjects) excrete disproportionately greater amounts of NE and its major extraneuronal metabolite, normetanephrine, than do controls. Depressed patients also show subsensitive neuroendocrine (growth hormone) and biochemical (inhibition of adenylate cyclase) responses to alpha 2-adrenergic agonists, suggesting that subsensitivity of nerve terminal alpha 2 autoreceptors may underlie the exaggerated plasma NE observed in response to various challenges in affective disorders. Future advances in brain imaging techniques and in the molecular biology of adrenergic receptor-coupled signal transduction systems offer promise for meaningful advances in our understanding of the pathophysiology of affective disorders.
Screening for familial defective apolipoprotein B-100 with improved U937 monocyte proliferation assay
Authors: Van den Broek A J , Hollaar L , Schaefer H I , Van der Laarse A , Schuster H , Defesche J C , Kastelein J J , Van 't Hooft F M 
Published: January 2020

Abstract
Frostegård et al. (J Lipid Res 1990;31:37-44) demonstrated that the proliferation of the human monocyte cell line U937 is critically dependent on the uptake of low-density lipoprotein (LDL) via the apo B, E (LDL) receptor, a characteristic that was used to detect patients with familial defective apolipoprotein B-100 (FDB). Here we applied this principle to develop a simple and reproducible assay for the detection of patients with functionally defective LDL. We added serum to U937 cells in cholesterol-free incubation medium and determined the increase in cell number after a 72-h incubation at 37 degrees C by using an electronic cell counter. Sera from 10 normolipidemic individuals and from 34 patients with type IIa hyperlipoproteinemia stimulated the growth of U937 cells in proportion to the exogenous cholesterol concentration (r = 0.83, P &lt; 0.001) and the LDL-cholesterol concentration (r = 0.81, P &lt; 0.001). However, sera from 16 patients with FDB stimulated less cell proliferation than did sera from patients with type IIa hyperlipoproteinemia with equal LDL-cholesterol concentrations. With a 15% reduction in growth as the cutoff value, this test had a sensitivity and specificity for diagnosis of FDB of 87.5% and 100%, respectively. The improved U937 monocyte proliferation assay can be used for screening hypercholesterolemic patients to detect individuals with functionally defective LDL.
The fitting, acceptance, and processing of standard curve data in automated immunoassay systems, as exemplified by the Serono SR1 analyzer
Authors: Daniels P B 
Published: January 2020

Abstract
Methodologies for establishing standard curves on automated immunoassay systems are dependent on the computational capability of the instrument, its throughput, and, perhaps, the technical experience of the user. Here, factors constraining this approach are discussed in general and the procedures used for the Serono SR1 analyzer are considered in detail. The SR1 is a moderate-throughput, fully automated immunoassay system capable of performing tests for (currently) 18 analytes. The associated user-generated standard curves are utilized not only for interpolation of results but also for evaluation of system performance. The choices of curve-fitting method, weighting of data, acceptance criteria, and outlier rejection are taken out of the hands of the user and are made a vital and integral part of the system. I justify this approach and demonstrate how a sensible mathematical approach must be tailored to the configuration of the instrument.
Enzymatic measurement of phosphatidylglycerol in amniotic fluid
Authors: Jones G W , Ashwood E R 
Published: January 2020

Abstract
We describe an enzymatic method for measuring phosphatidylglycerol (PG) at concentrations as low as 0.2 mumol/L in amniotic fluid. Amniotic fluid (1.5 mL) is centrifuged at 10,000g for 20 min to obtain a lamellar body pellet, which is reconstituted with 0.5 mL of buffer. The PG is measured by a two-step enzymatic scheme. Recovery studies demonstrated that the pellet contains &gt; 97% of the PG present in amniotic fluid. Between-run CVs were 28%, 5.7%, and 2.6% for amniotic fluid controls with means of 0.32, 3.9, and 10.7 mumol/L, respectively (n = 20). The enzymatic procedure was not significantly affected by blood, meconium, bilirubin, or other phospholipids. Lecithin/sphingomyelin ratio (n = 101) and fluorescence polarization (n = 127) compared with log(PG) showed correlation coefficients of 0.832 and -0.866, respectively. This test's ability to detect low concentrations of PG in amniotic fluid may make it a better predictor of fetal lung immaturity than previous methods.
Two-site enzyme immunoassay of cholesteryl ester transfer protein with monoclonal and oligoclonal antibodies
Authors: Mezdour H , Kora I , Parra H J , Tartar A , Marcel Y L , Fruchart J C 
Published: January 2020

Abstract
We developed a sandwich-type enzyme immunoassay to measure cholesteryl ester transfer protein (CETP) mass in human plasma. A specific monoclonal antibody (TP-4) that recognizes an epitope located in the C-terminal domain was used for antigen capture and an anti-CETP peptide antibody directed against the 290-306 residue was used for detection. Bound antibodies were revealed with an antibody-peroxidase conjugate specific for rabbit IgG. The presence of 10 mL/L Triton X-100 in the incubation buffer increased antigen exposure of CETP in plasma. The curves for CETP in standard plasma and partially purified CETP were parallel. This technique is rapid (results within 6 h), accurate, precise (mean intra- and interassay CVs 3.6% and 8.4%, respectively), and simple to perform. Assay sensitivity is at microgram concentrations, with a working range of 20-200 micrograms/L. In 40 normolipidemic healthy subjects, the mean CETP concentration in plasma was 1.1 +/- 0.4 mg/L. A strong correlation between CETP concentration and CETP activity (r = 0.91, n = 42) was observed. In plasma, the bulk of CETP was found in high-density lipoprotein fractions. Therefore, this assay may be a useful tool for investigations of CETP and its significance in relevant diseases.
Enzymes in diagnostics: achievements and possibilities of recombinant DNA technology
Authors: Kopetzki E , Lehnert K , Buckel P 
Published: January 2020

Abstract
We discuss, from an industrial point of view, the scope and possibilities of recombinant DNA technology for "diagnostic enzyme" production and application. We describe the construction of enzyme-overproducing strains and show how to simplify downstream processing, increase product quality and process profitability, improve diagnostic enzyme properties, and adjust enzymes to harsh assay conditions. We also consider some safety and environmental aspects of enzyme production. Other aspects of diagnostic enzymes that we cover are the facilitation of enzyme purification by attachment of short amino acid tails, the introduction of tails or tags for site-specific conjugation or oriented immobilization, the construction of bi- or multifunctional enzymes, and the production of enzyme-based diagnostic tests as demonstrated by the homogeneous immunoassay system of CEDIA tests. We use as examples of diagnostic enzymes glucose-6-phosphate dehydrogenase (EC 1.1.1.49), glucose oxidase (EC 1.1.3.4), alkaline phosphatase (EC 3.1.3.1), alpha-glucosidase (EC 3.2.1.20), pyruvate oxidase (EC 1.2.3.3), creatinase (EC 3.5.3.3), and beta-galactosidase (EC 3.2.1.23).
Multicomponent analysis for alkaline phosphatase isoenzyme determination by multiple linear regression
Authors: Tillyer C R , Rakhorst S , Colley C M 
Published: January 2020

Abstract
Alkaline phosphatase (EC 3.1.3.1) isoenzymes in serum may be determined by multicomponent analysis of the enzyme activities in the presence of multiple inhibitors. To determine inhibition coefficients of the isoenzymes, we used multiple linear regression analysis to compare alkaline phosphatase activities in the presence of known inhibitors with electrophoretically determined isoenzyme activities in plasma and serum samples. All possible combinations of exactly determined and overdetermined linear systems of inhibitors were ranked according to their prediction error to select an optimum set. The best multicomponent system for prediction included the use of levamisole, phenylalanine, and heat inhibition at 56 degrees C and 65 degrees C to determine bone, hepatic, intestinal, and placental isoenzymes. Consideration of the hepatic isoenzyme as liver and macromolecular fractions resulted in significantly worse predictions. Error analysis involving repeat determinations and a simplex optimization of the inhibition coefficients indicated that the inaccuracy of the comparison electrophoretic method may have been a major factor affecting poor isoenzyme prediction in some samples.
ELISA of IgG antibody to oxidized low-density lipoprotein: effects of blocking buffer and method of data expression
Authors: Craig W Y , Poulin S E , Nelson C P , Ritchie R F 
Published: January 2020

Abstract
We describe an ELISA for serum IgG antibodies against malondialdehyde-modified low-density lipoprotein (mLDL). Optimal antigen concentration, serum dilution, and dilution of enzyme-conjugated second antibody were 25 mg/L, 1:250, and 1:5000, respectively, when 5 g/L human serum albumin was used for blocking. When data were expressed as mLDL/LDL (the ratio of IgG binding to mLDL vs LDL), within-run and between-run CVs were 7.0% and 8.9%, respectively. Antibody concentrations expressed as mLDL/LDL or as mLDL-LDL (the difference between IgG binding to mLDL and to LDL) were higher in women with systemic lupus erythematosus (n = 20) than in controls (n = 20) (P &lt; 0.001). With bovine serum albumin or Superblock blocking buffers, only the mLDL-LDL data were significant. Thus, the choice of blocking agent and the method of data expression should be carefully considered when assaying IgG antibodies against mLDL.
Realistic modeling of clinical laboratory operation by computer simulation
Authors: Vogt W , Braun S L , Hanssmann F , Liebl F , Berchtold G , Blaschke H , Eckert M , Hoffmann G E , Klose S 
Published: January 2020

Abstract
An important objective of laboratory management is to adjust the laboratory's capability to the needs of patients' care as well as economy. The consequences of management may be changes in laboratory organization, equipment, or personnel planning. At present only one's individual experience can be used for making such decisions. We have investigated whether the techniques of operations research could be transferred to a clinical laboratory and whether an adequate simulation model of the laboratory could be realized. First we listed and documented the system design and the process flow for each single laboratory request. These input data were linked by the simulation model (programming language SIMSCRIPT II.5). The output data (turnaround times, utilization rates, and analysis of queue length) were validated by comparison with the current performance data obtained by tracking specimen flow. Congruence of the data was excellent (within +/- 4%). In planning experiments we could study the consequences of changes in order entry, staffing, and equipment on turnaround times, utilization, and queue lengths. We conclude that simulation can be a valuable tool for better management decisions.
Simultaneous macroamylasemia and macrolipasemia
Authors: Zaman Z , Van Orshoven A , Mariën G , Fevery J , Blanckaert N 
Published: January 2020

Abstract
The first case of the simultaneous presence of macroamylasemia and macrolipasemia in a patient with gluten enteropathy (celiac disease) is described. Both macroenzymes were formed by association of polyclonal IgA with amylase and lipase. Both macroenzymes had molecular masses &gt; 300 kDa.
Application of the clearance concept to hyponatremic and hypernatremic disorders: a phenomenological analysis
Authors: Shoker A S 
Published: January 2020

Abstract
The kidney and its response to the antidiuretic hormone (ADH) are the principal protective mechanisms necessary to maintain a normal plasma tonicity (osmolality). Hence, determination of the response of the ADH-renal axis to an abnormal plasma tonicity is an important step to understanding water homeostasis. Determination of free water clearance is the most direct clinical method to measure the ability of the kidney to reabsorb or excrete water; it can be used as a sensitive method to study water metabolism, describing the abnormal water homeostasis in simple quantitative terms. A positive electrolyte-free water clearance denotes the excretion of excess free water. A negative electrolyte-free water clearance indicates reabsorption of excess free water. During hypertonicity, an increased concentration of ADH enhances renal reabsorption of free water. With diminished ADH secretion and normal renal function, a substantial volume of free water is cleared in response to hypotonic stimuli. A positive free water clearance &gt; 0.4 L/day in hypertonic conditions or a negative free water clearance during hypotonicity confirms an abnormal ADH-renal axis response.
Comparable detection of acute myocardial infarction by creatine kinase MB isoenzyme and cardiac troponin I
Authors: Adams J E , Schechtman K B , Landt Y , Ladenson J H , Jaffe A S 
Published: January 2020

Abstract
Although measurement of cardiac troponin I (cTnI) is, in some situations, more specific for detection of cardiac injury than is measurement of the MB isoenzyme of creatine kinase (MBCK), its sensitivity and specificity relative to MBCK for detection of myocardial infarction has not been established. Accordingly, we studied prospectively 199 consecutive patients admitted to the coronary care unit. Values of MBCK and cTnI mass were determined in all samples. Of the 188 patients admitted with a suspicion of acute myocardial ischemia, 89 were diagnosed as having an acute myocardial infarction on the basis of the patterns of MBCK values. Eighty-six of these patients also had increased cTnI (concordance, 96.6%); three did not. Of the patients diagnosed as without infarction, five with unstable angina and symptoms in the day(s) prior to admission had increased cTnI, for a cTnI specificity of 94.9%. Receiver operating characteristic curve analysis indicated that cTnI and MBCK had statistically indistinguishable diagnostic accuracies for the detection of acute myocardial infarction.
Improved HPLC-radioimmunoassay for quantifying angiotensin II in plasma
Authors: Voelker J R , Cobb S L , Bowsher R R 
Published: January 2020

Abstract
To measure the octapeptide angiotensin II (Ang II) in plasma, we developed a sensitive, specific assay that interfaces solid-phase extraction, HPLC, and RIA. A reversed-phase HPLC system involving isocratic elution at 38 degrees C with a volatile mobile phase of acetonitrile and the ion-pairing reagent heptafluorobutyric acid produced baseline separation of angiotensin peptides. Ang II was collected as a single fraction, concentrated by evaporation to dryness, and measured by RIA after resuspension in RIA buffer. Even including column washing between sample injections to prevent carryover of plasma constituents, two plasma extracts could be processed per hour by HPLC. Assay validation experiments demonstrated &lt; 2% cross-reactivity with Ang II-related peptides; a 75% recovery from plasma at physiological concentrations of Ang II; intra- and interassay precision (CVs) of 6.2% and 10.3%, respectively; and a lower limit of quantification of 1.3 ng/L. Two clinical protocols designed to measure plasma Ang II concentration under basal and stimulated conditions confirmed the utility of the assay.
Indoxyl-beta-D-glucuronide, the primary emitter of low-level chemiluminescence in plasma of hemodialysis patients
Authors: Agatsuma S , Sekino H , Nagoshi T , Watanabe H 
Published: January 2020

Abstract
Characteristic light emission induced by the oxidation of hydroxyl radicals has been found in plasma of hemodialysis patients (Agatsuma et al., Clin Chem 1992;38:48-55). We purified a primary emitter, a chemiluminescent component peaking at 430 nm, by anion-exchange chromatography and reversed-phase HPLC. By using proton nuclear magnetic resonance and authentic indoxyl compounds, we determined the primary emitter to be indoxyl-beta-D-glucuronide. Absorption and fluorescence spectra of the purified sample coincided well with those of authentic indoxyl-beta-D-glucuronide, as did the peak in the chemiluminescence emission spectrum. Retention time of the purified sample on reversed-phase HPLC, measured by fluorescence, was also in accordance with that of indoxyl-beta-D-glucuronide. To our knowledge, this is the first identification of a primary emitter of low-level chemiluminescence from a biological source.