bluuebunny/crossref_metadata_2025_split huggingface dataset:
Two-dimensional gel electrophoresis detects prostate-specific antigen–α1-antichymotrypsin complex in serum but not in prostatic fluid
Authors: Qian Yi , Sensibar Julia A , Zelner David J , Schaeffer Anthony J , Finlay Judith A , Rittenhouse Harry G , Lee Chung 
Published: January 2020

AbstractWe investigated the interaction between prostate-specific antigen (PSA) and 1-antichymotrypsin (ACT) in prostatic secretions, identifying PSA and ACT in human serum, prostatic fluid, and seminal plasma by two-dimensional gel electrophoresis (2-D PAGE). Both PSA and ACT were detected in all three body fluids, but PSA-ACT complex was detected only in serum. Moreover, the 2-D PAGE Western blot staining profile for ACT from serum differed from that for prostatic fluid or seminal plasma. Incubation of prostatic fluid with purified ACT led to formation of PSA-ACT complex. Incubation of prostatic fluid with purified PSA, however, failed to form the complex, suggesting that the ACT in prostatic fluid was inactive or inhibited. Given that physiological concentrations of zinc inhibited the formation of PSA-ACT complex, we consider zinc a possible physiological inhibitor of the formation of the PSA-ACT complex. These results indicate that the failure to detect the PSA-ACT complex in prostatic fluid could be related to the inactivation of ACT, the presence of inhibitors (e.g., zinc), or simply the PSA:ACT ratio in the fluid.
Fluid elements—a concept for automation of diagnostic tests
Authors: Van Damme Henk , Van Velthoven Thea , Kaelen Erik , Pelssers Eduard 
Published: January 2020

Abstract
Constructs consisting of a channel, a membrane, and an absorber are designed for autonomously carrying out various liquid-handling functions of analytical tests. These so-called fluid elements can be used to set up various circuits for conducting several kinds of analytical tests. To demonstrate the feasibility of this concept, we constructed such a circuit and used it to perform, with two handling steps, an ELISA of hepatitis B surface antigen. The detection limit of the assay was comparable with those of state-of-the-art ELISAs for screening blood, and results could be obtained within a total test time of 20 min. We anticipate that this concept of automation may also serve as a basis for new, highly simplified immunoanalyzers.
Whole-blood test for total cholesterol by a self-metering, self-timing disposable device with built-in quality control
Authors: Law Wai Tak , Doshi Sonal , McGeehan John , McGeehan Susan , Gibboni David , Nikolioukine Yuri , Keane Robert , Zheng Jennifer , Rao Jagan , Ertingshausen Gerhard 
Published: January 2020

AbstractA whole-blood test for total cholesterol has been developed that is performed in a low-cost disposable flow device without user intervention (after sample addition). The device meters the sample, separates plasma from erythrocytes, and precisely times plasma flow into various reagent compartments, including a quality-assurance chamber. Test results are displayed as a well-defined and easily readable color bar. A quality-control window attests to the integrity of the test components. Here, we describe the assembly and individual functions of the device and report its performance characteristics. Precision and accuracy studies in four clinical studies at independent locations yielded total imprecision of &lt;5% and an average bias of 1.35% vs the Abell–Kendall method.
Serum ionized magnesium in chronic alcoholism: is it really decreased?
Authors: Hristova Elena N , Rehak Nadja N , Cecco Stacey , Ruddel Mark , Herion David , Eckardt Michael , Linnoila Markku , Elin Ronald J 
Published: January 2020

Abstract
Chronic alcoholism is associated with a marked deficit in total magnesium (tMg). However, little is known about the status of the physiologically active form, ionized magnesium (iMg). We assessed serum iMg (measured with two ion-selective electrodes, AVL 988-4 and NOVA CRT) and tMg concentrations in chronic alcoholics at admission (n = 31) and after abstinence (n = 13) and compared these results with those for a control group (n = 40). At admission, the tMg and NOVA iMg concentrations in alcoholics (0.78 ± 0.020 and 0.38 ± 0.016 mmol/L, respectively) were significantly less (P &lt;0.001) than in the controls (0.85 ± 0.008 and 0.50 ± 0.006 mmol/L). The AVL iMg results, however, did not differ significantly between the two groups: 0.53 ± 0.013 vs 0.56 ± 0.006 mmol/L, respectively (P &gt;0.05). The mean iMg between the two analyzers differed significantly in both groups (P &lt;0.001). After 3 weeks of abstinence, the alcoholics showed a significant increase in tMg (P &lt;0.001) and in both NOVA and AVL iMg values (P &lt;0.01 for each). tMg concentrations were positively correlated with the AVL iMg values in both alcoholics and controls but correlated positively with the NOVA iMg results only in the controls. Thus, the altered status of iMg is instrument-dependent, and the usefulness of the measurement in alcoholics is yet to be determined.
Automated liquid-chromatographic analyzer used for toxicology screening in a general hospital: 12 months’ experience
Authors: Sadeg Nouredine , François Gilles , Petit Brigitte , Dutertre-Catella Hélène , Dumontet Michel 
Published: January 2020

Abstract
We evaluated the clinical utility of an automated HPLC system (Remedi, Bio-Rad) for identification of drugs and metabolites in biological fluids. Serum or urine or both from 354 consecutive cases of poisoning were analyzed by the system and by a set of fluorescence polarization immunoassay (FPIA, Abbott) and thin-layer chromatographic (TLC) procedures. Antidepressants and most phenothiazines were recognized by the new system. Comparison of Remedi results with final clinical diagnoses yielded diagnostic specificity and sensitivity of 80% and 90%, respectively. Remedi detected 26 additional compounds that were neither reactive in the immunoassay screening tests nor detected by TLC procedures. Because the Remedi expands the range of drugs covered by the immunoassays and provides a rapid, preliminary report in emergency situations, we conclude that this system can be a useful complementary technique in the clinical toxicology laboratory. Although urine toxicological screening seemed adequate for a good toxicological report, blood analysis allows extra toxicokinetic data such as blood concentrations and half-life estimations.
Immunoassay of insulin-like growth factor binding protein-1
Authors: Khosravi M Javad , Diamandi Anastasia , Mistry Jehangir 
Published: January 2020

AbstractAccurate measurement of insulin-like growth factor (IGF) binding protein-1 (IGFBP-1) is important for precise definition of its physiological roles and potential diagnostic values. Because altered phosphorylation results in altered IGFBP-1 immunoreactivity, current assays may significantly underestimate or fail to detect physiological changes in the IGFBP-1 concentrations. We developed three ELISAs (ELISA 1–3) using a common capture but three different detection antibodies. IGFBP-1 in serum, synovial fluid (SF), cerebrospinal fluid (CSF), and amniotic fluid (AF) were measured before and after treatment with alkaline phosphatase (ALP). Among the methods, only ELISA-1 was unaffected by IGFBP-1 phosphorylation and generated identical results before and after ALP treatment. The serum and SF values by ELISA-2 and -3 were lower by ∼4- to 10-fold, but increased after ALP treatment to within 66–98% of those by ELISA-1. The medians in AF, and to a lesser extent in CSF, by all methods were similar and did not change significantly after dephosphorylation. ELISA-1 showed excellent correlation with ELISA-2, ELISA-3, and a commercial IGFBP-1 IRMA only after ALP-treated samples were analyzed by the comparative methods. ELISA-1 is highly specific for IGFBP-1 and demonstrated acceptable analytical performance characteristics.
Prostate-specific antigen: characterization of epitopes by synthetic peptide mapping and inhibition studies
Authors: Corey Eva , Wegner Sandra K , Corey Michael J , Vessella Robert L 
Published: January 2020

Abstract
To improve our understanding of the prostate-specific antigen (PSA) antigenic regions, we studied the association targets of one anti-PSA polyclonal antibody and 10 anti-PSA monoclonal antibodies (mAbs). We also examined the ability of the mAbs to inhibit PSA enzymatic activity and block the association of PSA with α1-antichymotrypsin (ACT). Linear epitope mapping with a polyclonal antibody indicated the presence of six major antigenic regions in PSA. Examination of the panel of mAbs established that three of them bind to linear epitopes. Five of the mAbs inhibited &gt;90% of PSA enzymatic activity. However, inhibition of PSA enzymatic activity and hindrance of PSA-ACT association by mAbs cannot be used to predict whether the mAbs bind to free PSA, the PSA-ACT complex, or both. Some of the mAbs may block PSA-ACT association through peripheral occlusion of the binding site, or through induction of conformational changes in PSA.
Computer-aided identifications of thin-layer chromatographic patterns in broad-spectrum drug screening
Authors: Siek Theodore J , Stradling Corey W , McCain Marc W , Mehary Timothy C 
Published: January 2020

Abstract
We have developed a systematic thin-layer chromatographic (TLC) technique for detecting and identifying drugs and drug metabolites on 10-cm-long silica-gel plates with organic binder (fluorescent indicator); a computer program (SPOT CHEK) assists in matching the data from a particular chromatogram with those obtained for known drugs recovered from serum, urine, or other specimens. The plates are developed in a single mobile phase. Visualization and detection reagents used to characterize unknowns include fluorescamine, ferric chloride/perchloric acid/nitric acid, Dragendorff, Marquis, Mandelin, and iodinated Dragendorff solutions, 254 nm ultraviolet light, and vapor from chlorine or hydrochloric acid. Detection limits of 5–200 ng per sample spot were obtained for drugs in the database. The computer program database is based on nine reaction responses plus the plate zone locations for 243 drug substances but requires entry of only one TLC property to generate a matching list. We ran the program with an IBM-compatible 386/486 PC using an MS-DOS operating system (version 6.2).
Effects of reducing reagents and temperature on conversion of nitrite and nitrate to nitric oxide and detection of NO by chemiluminescence
Authors: Yang Fan , Troncy Eric , Francœur Martin , Vinet Bernard , Vinay Patrick , Czaika Guy , Blaise Gilbert 
Published: January 2020

Abstract
To measure the concentration of nitrites and nitrates by chemiluminescence, we examined the efficiency of five reducing agents [V(III), Mo(VI) + Fe(II), NaI, Ti(III), and Cr(III)] to reduce nitrite (NO2−) and (or) nitrate (NO3−) to nitric oxide (NO). The effect of each reducing agent on the conversion of different amounts of NO2− and (or) NO3−(100–500 pmol, representing concentrations of 0.4 to 2 μmolar) to NO was determined at 20 °C for NO2− and at 80 °C for NO3−. The effect of temperature from 20 to 90 °C on the conversion of a fixed amount of NO2− or NO3− (400 pmol or 1.6 μmolar) to NO was also determined. These five reducing agents are similarly efficient for the conversion of NO2− to NO at 20 °C. V(III) and Mo(VI) + Fe(II) can completely reduce NO3− to NO at 80 °C. NaI and Cr(III) were unable to convert NO3− to NO. Increased temperature facilitated the conversion of NO3− to NO, rather than that of NO2− to NO. We evaluated the recovery of NO2− and NO3− from plasmas of pig and of dog. Recovery from plasma of both animals was reproducible and near quantitative.
Vitamine—vitamin. The early years of discovery
Authors: Rosenfeld Louis 
Published: January 2020

AbstractIn 1905, Cornelius Adrianus Pekelharing found that animals fed purified proteins, carbohydrates, fats, inorganic salts, and water would thrive only if small amounts of milk were added to the diet. He concluded that the milk contained some unrecognized substance that in very small quantities was necessary for normal growth and maintenance. In 1911, Casimir Funk isolated a concentrate from rice polishings that cured polyneuritis in pigeons. He named the concentrate “vitamine” because it appeared to be vital to life and because it was probably an amine. Although the concentrate and other “accessory food substances” were not amines, the name stuck, but the final “e” was dropped. In 1913 two groups discovered a “fat-soluble” accessory food substance. Initially believed to be a single vitamin, two separate factors were involved. One, effective against xerophthalmia, was named vitamin A; the other, effective against rickets, was named vitamin D. The factor that prevented scurvy was isolated in 1928. Known as “water-soluble C,” it was renamed ascorbic acid.
Quantitative polymerase chain reaction-based homogeneous assay with fluorogenic probes to measure c-erbB-2 oncogene amplification
Authors: Gelmini Stefania , Orlando Claudio , Sestini Roberta , Vona Giovanna , Pinzani Pamela , Ruocco Luca , Pazzagli Mario 
Published: January 2020

AbstractWe describe a PCR-based assay for determining c-erbB-2 oncogene amplification in breast cancer in which we use the TaqManTM system. Two fluorogenic probes anneal to the target between primers for c-erbB-2 and β-globin genes and contain both a reporter dye (6-carboxy-fluorescein) and a quencher dye (6-carboxy-tetramethyl-rhodamine). During the extension phase of the PCR cycle, the 5′→3′ exonuclease activity of Taq polymerase cleaves the hybridized fluorogenic probe, resulting in an increase of fluorescence emission of the reporter dye that is quantitative for the amount of PCR product and, under appropriate conditions, for the amount of template. Assay performance showed adequate precision and a lower detection limit and good correlation with the results obtained in the same samples by a competitive PCR assay (n = 25, r = 0.94, P &lt;0.01). This homogeneous assay is time-saving, avoids usually cumbersome postamplification procedures (that can be additional sources of inaccuracy and contamination), and seems suitable for determination of c-erbB-2 oncogene amplification in tumor specimens.
Construction of cDNA bank from biopsy specimens for multiple gene analysis of cancer
Authors: Ishikawa Takashi , Ichikawa Yasushi , Miura Yasuhiko , Momiyama Mobuyoshi , Keller Cylia , Koo Kenneth , Akitaya Tatsuo , Shimada Hiroshi , Mitsuhashi Masato 
Published: January 2020

AbstractWe constructed a “cDNA bank” of human colorectal cancer and surrounding normal tissues with our unique mRNA assay system. Total nucleic acids extracted from patients’ tissues were applied to 96-well plates, where poly(dT) sequences of oligonucleotides were immobilized. After hybridization, the cDNA was reverse-transcribed on the plate with the captured mRNA as a template, followed by synthesis of double-stranded (ds) cDNA. The resulting sense cDNA was removed from the plate, then used in PCR for analysis of various genes. The sense strand of the cDNA was repeatedly synthesized by using the immobilized antisense cDNA as a template even from plates used once and stored at 4 °C for as long as 6 months. Furthermore, the results of PCR could be easily compared among different specimens if the same amount of total mRNA were applied to the plate for the ds cDNA synthesis. This demonstrated that the cDNA bank constructed from clinical materials provides almost unlimited supplies of cDNA for multiple gene analysis of cancer.
Automation of human sperm cell analysis by flow cytometry
Authors: Ferrara Fulvio , Daverio Rita , Mazzini Giuliano , Bonini Pierangelo , Banfi Giuseppe 
Published: January 2020

AbstractSemen sample analysis is routinely performed by microscopical evaluation and manual techniques by laboratory operators; the analysis is affected by a wide imprecision related to variability among observers, influencing its clinical validity. Our aim was to automate sperm analysis with the use of flow cytometry for evaluation of cell counts and typing and with the use of a new membrane-permeant nucleic acid stain for evaluation of sperm viability. Statistical analysis of the comparison between manual and automated methods for sperm counts was performed by the Bland and Altman method; the mean difference was 0.243 × 106 sperms/mL. The precision of the flow cytometric analysis was evaluated with whole sperm; the between-run CV was 7.5% and the within-run CV was 2.5%. Data observed suggest that flow cytometric sperm analysis, with high precision and accuracy and low costs, can be proposed for routine use in clinical laboratories.
Three generations of high-density lipoprotein cholesterol assays compared with ultracentrifugation/dextran sulfate–Mg2+ method
Authors: Harris Neil , Galpchian Vartouhi , Thomas Jennifer , Iannotti Elizabeth , Law Terence , Rifai Nader 
Published: January 2020

AbstractWe report on the analytical performance of three generations of HDL-cholesterol assays: phosphotungstic acid/Mg2+, Spinpro®, and a homogeneous method, N-geneous™. The run-to-run imprecision (CV) of all assays was ≤4.9%, and all results correlated highly with those of a modified reference procedure (r ≥0.96). At triglycerides concentrations &lt;4000 mg/L, these field methods showed an acceptable systematic error (y = 1.12x − 47, 1.05x − 23, and 0.96x + 8 for the phosphotungstate, Spinpro, and N-geneous assays, respectively), and the total error of the field methods met the current National Cholesterol Education Program (NCEP) performance goal of ≤22%. Regression analyses of results for samples with triglycerides &gt;4000 mg/L produced the following results for the above respective assays: y = 1.08x − 4.2, 1.02x + 3.6, and 0.85x + 108. In this hypertriglyceridemic group, only the N-geneous assay (at an HDL-cholesterol content of 240 mg/L) had a total error (35%) that exceeded the NCEP limit. Bilirubin and ascorbate produced a negative interference with the phosphotungstate and Spinpro assays but had little effect on the N-geneous assay.
Gastric tubes, meals, acid, and analysis: rise and decline
Authors: Rosenfeld Louis 
Published: January 2020

Abstract
The stomach tube was first used to administer food and medication or to remove poisonous substances. Later, it served to aspirate the stomachs of patients with gastric retention. Chemical analysis of stomach contents after a meal was first suggested in 1871 and quickly became an important laboratory procedure as various test-meal stimuli and more flexible tubes were developed. Quantitative estimations of free and total acidity were made by titration with 0.01 mol/L sodium hydroxide and specific indicators. Pentagastrin has supplanted secretagogues such as histamine and betazole; meal stimulation, tubeless tests, and other tests of gastric secretion are no longer used clinically. Tests of gastric acid secretion have been used in the diagnosis of upper gastrointestinal lesions and to help select the type of surgical procedure for gastric and duodenal ulcers, but the tests have decreased in importance because of their limited diagnostic sensitivity and specificity. Today, fiberoptic endoscopy is replacing gastric analysis as well as radiologic examination.
Quality: the next six months
Authors: Laessig Ronald H , Ehrmeyer Sharon S 
Published: January 2020

Abstract
The eclectic mix of participants in the forum had a surprisingly singular focus when it came to the topic of quality in clinical laboratories. All sensed that the time is right for a transition from laws, rules, and inspections to a true quality-based system. Such a system can achieve the goals, implicit and explicit, that are the rationale for the multiplicity of regulations affecting today’s laboratories. A true quality-based system has great potential benefits to laboratories, regulators, and manufacturers, and ultimately to our true customers, the patients. The benefits include lower costs, superior products, and better test results; in short, better patient care. This transition will be possible only through formation of a “Quality Alliance,” composed of those skilled in the “theory” of quality—laboratory personnel, manufacturers, and regulators, acting as one to implement the quality system. The Quality Alliance requires a team of individuals with different skills, aligned as one, for the purpose of achieving a common goal. On the basis of views expressed in this Forum, our collective future will be defined by the evolving Quality Alliance, an alliance focused on true quality systems in clinical laboratories.
Initial evaluation of cystatin C measurement by particle-enhanced immunonephelometry on the Behring nephelometer systems (BNA, BN II)
Authors: Finney Hazel , Newman David J , Gruber Walter , Merle Peter , Price Christopher P 
Published: January 2020

AbstractSerum cystatin C has been suggested as a new marker of glomerular filtration rate (GFR). We describe a fully automated and rapid particle-enhanced nephelometric immunoassay (PENIA) for measuring serum cystatin C on the Behring nephelometer systems (BNA, BN II). Each sample is analyzed in 6 min with as many as 75 samples per batch. The assay covers the range 0.23–7.25 mg/L, up to seven times the upper limit of normal. The intra- and interassay imprecision are &lt;3.3% and &lt;4.5%, respectively. There is absolute linearity across the assay range (r2 = 0.997), with analytical recovery by cystatin C addition between 95% and 109% (mean 102%). Hemoglobin (≤8.0 g/L), bilirubin (≤488 μL), triglycerides (≤23 mmol/L), rheumatoid factor (≤2000 kIU/L), and myeloma paraprotein (≤41 g/L) do not interfere with the assay. This assay agreed well with an in-house particle-enhanced turbidimetric immunoassay (PETIA) (mean difference = 1.73 ± 2.10) and a commercial PETIA (mean difference = 1.13 ± 0.86). This is a new assay by which cystatin C may be effectively used as a marker of GFR estimation.
Gastrointestinal absorption, tissue retention, and urinary excretion of dietary aluminum in rats determined by using 26Al
Authors: Jouhanneau Philippe , Raisbeck Grant M , Yiou Françoise , Lacour Bernard , Banide Hélène , Drüeke Tilman B 
Published: January 2020

Abstract
We used accelerator mass spectrometry (AMS) and 26Al to study the plasma concentration, urinary excretion, and retention in bone, brain, and liver of a single dose of a dietary concentration of aluminum ingested either with or without citrate by 2-month-old Wistar rats. In the absence of citrate, cumulative urinary excretion and skeleton retention were each ∼0.05% of the total 26Al dose ingested. 26Al retention in brain and liver were ∼4 × 10−8 and 2 × 10−6, respectively. Concomitant citrate intake increased these median values by about two- to fivefold, although this factor was highly variable in individual rats. Independent of citrate administration, 90% of the26Al excreted in urine (measured cumulatively over 30 days) was excreted within the first 48 h. Uptake by bone was rapid (∼1 h) and permanent over the 30-day duration of the experiment.