bluuebunny/crossref_metadata_2025_split huggingface dataset:
Serum concentrations of carboxyl-terminal propeptide of type I procollagen, amino-terminal propeptide of type III procollagen, cross-linked carboxyl-terminal telopeptide of type I collagen, and their interrelationships in schoolchildren
Authors: Crofton Patricia M , Wade Jean C , Taylor Mervyn R H , Holland Celia V 
Published: January 2020

AbstractWe report pediatric age- and sex-specific 95% reference intervals for procollagen type I C-terminal propeptide (PICP), the cross-linked C-terminal telopeptide of type I collagen (ICTP), and procollagen type III N-terminal propeptide (P3NP), measured in plasma from 302 schoolchildren (156 boys, 146 girls) ages 4–19 years. All three markers displayed a significant variation with age (ANOVA P ≤0.0015). PICP showed no detectable increase during adolescence for either sex, but decreased towards adult concentrations after the age of puberty, with an earlier decrease for girls than for boys (P &lt;0.01). ICTP and P3NP both increased in pubertal-aged children (P &lt;0.05), with an earlier increase in girls than in boys (P &lt;0.05), before decreasing towards adult concentrations (P &lt;0.01). All three collagen markers were highly correlated with one another (P &lt;0.001). The patterns observed mirrored the childhood growth curve and reflected the high turnover of bone and soft tissue during childhood growth.
Thiocyanate in smokers interferes with the Nova magnesium ion-selective electrode
Authors: Rehak Nadja N , Cecco Stacey A , Niemela Julie E , Elin Ronald J 
Published: January 2020

Abstract
Thiocyanate found in serum ordinarily is the metabolite of cyanide that is inhaled with tobacco smoke and ingested with cyanogenic foods. We investigated the effect of the thiocyanate ion (SCN−) on the ionized magnesium (iMg) and ionized calcium (iCa) results determined with the AVL and Nova magnesium and calcium ion-selective electrodes (ISEs). We analyzed saline and pooled serum with added SCN−, and serum from apparently healthy nonsmokers (n = 20) and smokers (n = 20). The mean (and range) of the measured serum SCN− concentration was 0.019 (0.008–0.046) mmol/L for nonsmokers and 0.077 (0.020–0.138) mmol/L for smokers. Only the Nova iMg results decreased with increasing SCN−concentration, and the change was dependent on the baseline iMg concentration. In the absence of Mg, SCN− decreased the voltage response of the Nova Mg ISE to calcium ions. At apparently normal serum iMg and iCa concentrations, the interference by SCN− appeared to be equimolar (iMg = −1.04 × SCN− + 0.52). Thus, the serum SCN− commonly found in smokers causes a significant (P &lt;0.0001) decrease in the Nova iMg results.
New tools for laboratory design and management
Authors: Truchaud Alain , Le Neel Tanguy , Brochard Hugues , Malvaux Sophie , Moyon Marine , Cazaubiel Murielle 
Published: January 2020

Abstract
The clinical laboratory is changing from a place of activity based on sample analysis to an in vitro diagnostic network. To convince our team, partners, and administrators, we need new comprehensive tools to define a strategy with limited risk of failure or conflicts. Specific quality goals should be established before choosing automated tools for sample handling, analytical systems, laboratory information systems, communication systems, or advanced technologies. A system approach maps and simplifies the process, based more on a functional study than on classical disciplines. A customer–supplier approach establishes the requirements between partners either inside or outside the laboratory. The quality system must be a management tool, linking samples, tasks, information, and documents. Quantitative simulation modeling explores different automation alternatives and their impact on laboratory workflow. Finally, integration of results in interactive semirealistic simulation tools for laboratory design or reengineering can be used as communications tools to involve laboratory professionals in the change of their practice.
ProC® Global: the first functional screening assay for the complete protein C pathway
Authors: Dati Francesco , Hafner Gerd , Erbes Hilde , Prellwitz Winfried , Kraus Michael , Niemann Frank , Noah Michael , Wagner Carola 
Published: January 2020

Abstract
In clinical practice, venous thromboembolic complications are much more frequent than bleeding disorders. In fact, disturbances within the protein C pathway due to coagulation factor V (FV) Leiden mutation and deficiency of protein C or protein S are the most frequent abnormalities in hereditary thrombophilia. Furthermore, acquired dysfunctions of the protein C system may predispose the single individual to an increased thrombotic risk. A routine-suited screening assay that would allow the monitoring of the proper interplay of factors in the protein C pathway could add an important factor to the basic coagulation profile. This consists of the prothrombin time and of the activated partial thromboplastin time, which currently allow only a screening for increased risk for bleeding but not for venous thromboembolism. A new functional screening test for the protein C system such as the presented ProC® Global should therefore facilitate detection of FV Leiden as well as deficiency of protein C and protein S. The results of the present evaluation indicate that ProC Global is highly sensitive to activated protein C resistance/FV Leiden (100%) and protein C deficiency (90%) and sensitive to protein S deficiency (63%). Furthermore, the assay gives a quantitative measure of the net potential of the protein C pathway in relation to the intrinsic procoagulant system. The use of this assay for a prospective assessment of thromboembolic risk is the subject of current studies.
In vitro effects of a novel hemoglobin-based oxygen carrier on routine chemistry, therapeutic drug, coagulation, hematology, and blood bank assays
Authors: Callas Demetra D , Clark Terri L , Moreira Paulo L , Lansden Cara , Gawryl Maria S , Kahn Stephen , Bermes Edward W 
Published: January 2020

Abstract
Red blood cell (RBC) replacement solutions are being developed as alternatives to allogeneic RBC use in blood transfusions in the treatment of massive trauma, to achieve hemodynamic stability during elective surgery, and to increase oxygen-carrying capacity in anemia. Hemoglobin-based oxygen carrier (HBOC)-201 (Biopure Corp.) is a purified, sterile, isosmotic glutaraldehyde-polymerized bovine hemoglobin. Because this product is acellular, blood components containing this substance appear hemolyzed. This study reports on the interferences produced by the presence of HBOC-201 in a variety of clinical assays. This product was added in vitro at concentrations up to 60 g/L (6.0 g/dL) to normal human serum, plasma, or whole blood before testing for serum chemistries, coagulation profiles, and hematology and blood bank assays. In addition, a set of normal human sera containing HBOC-201 was supplemented with various therapeutic drugs and assayed for these agents. The results of these studies demonstrate that the presence of HBOC-201 in blood components does not result in significant analytical interference that would be of concern with many clinical assays at HBOC-201 concentrations encountered during routine clinical use of this RBC replacement solution in patients.
A laboratorian’s perspective on evaluation and implementation of new laboratory tests
Authors: O’Kane Dennis J , Ebert Thomas A , Hallaway Brenda J , Roberts Steven G , Bhuiyan Abul K M J , Tenner Kathleen S 
Published: January 2020

Abstract
New assay development should be directed toward answering fundamental clinical questions. Caveats that must be considered before initiating assay development projects are: New assays should allow the clinician to interact with and treat a patient more effectively, thereby improving medical outcome; and new assays should facilitate recapture of system resources, enabling cost savings or reinvestment of resources. Defining the clinical questions and consideration of the caveats permit a means of prioritizing assay development activities. Laboratorians are faced with evaluating several types of development activities that lead to assay implementation in routine clinical testing. Assays can be prioritized for up-grading to newer cost-effective technologies, provided the changes maintain or improve analytical and clinical performance. Predicting which research assay will have future value is difficult when clinical performance is not fully validated. However, such assay development has the greatest potential for changing the delivery of healthcare by a clinician.
Validation by isoelectric focusing of the anion-exchange isotransferrin fractionation step involved in determination of carbohydrate-deficient transferrin by the CDTect assay
Authors: Arndt Torsten , Hackler Rolf , Kleine Tilman O , Gressner Axel M 
Published: January 2020

Abstract
Serum concentration of carbohydrate-deficient transferrin (CDT) is used for laboratory diagnosis of chronic alcohol abuse. Using isoelectric focusing for validation of the initial isotransferrin fractionation step involved in the determination of CDT by the CDTect assay, we found a complete in vitro iron saturation of transferrin and sufficient stability of the transferrin iron load during column passage; effective separation of non-CDT-isotransferrins and CDT-isotransferrins at the microcolumns; partial coelution of trisialo-Fe2-transferrin, which did not significantly affect CDT measurement; partial retention of CDT-isotransferrins, especially disialo-Fe2-transferrin, which may cause falsely negative results for CDT at the upper reference limits; good precision of the isotransferrin fractionation step; and no significant effects of low concentrations of serum protein and transferrin. We strongly urge standardization of CDT analysis and suggest isoelectric focusing for validation of CDT analysis methods and verification of odd results.
Chemical mismatch cleavage combined with capillary electrophoresis: detection of mutations in exon 8 of the cystathionine β-synthase gene
Authors: Ren Jicun , Ulvik Arve , Refsum Helga , Ueland Per Magne 
Published: January 2020

Abstract
Mutation detection by chemical mismatch cleavage (CMC) is based on the chemical modification and cleavage at the site of mismatched C or T in heteroduplexes, using hydroxylamine or osmium tetroxide (OsO4) as chemical probes. In the present study, we evaluated CMC in combination with capillary electrophoresis (CE) by determining the common T833C and G919A mutations in exon 8 of the cystathionine β-synthase gene in heterozygous and homozygous samples. A 186-bp fragment encompassing exon 8 was amplified by PCR with both primers labeled with 5′-fluorescein. Labeled single strands of 40 and 61 nucleotides (nt) were formed from the coding strand of the T833C sample and non-coding strand from the G919A sample, respectively. These single-stranded DNA (ssDNA) products were analyzed under denaturing conditions by CE with short-chain linear polyacrylamide as the sieving matrix and were detected by laser-induced fluorescence (LIF), using a sensitive, one-channel sheath-flow detector. The CE-LIF format afforded relatively high resolution of ssDNA (down to 1 nt), precise size assessment of CMC products, sensitive detection with small sample requirements, and fast analysis. Thus, CMC combined with CE-LIF is suitable for screening of known mutations, giving expected CMC products, but will also detect unknown mutations, the locations of which are indicated by the fragment sizes.
Analytical and clinical performance characteristics of Tandem-MP Ostase, a new immunoassay for serum bone alkaline phosphatase
Authors: Broyles Dennis L , Nielsen Randall G , Bussett Elizabeth M , Lu W Douglas , Mizrahi Isaac A , Nunnelly Patricia A , Ngo Tram A , Noell Julia , Christenson Robert H , Kress Barry C 
Published: January 2020

AbstractThe performance characteristics of the Tandem®-MP Ostase® assay, a new microplate immunoassay for bone-specific alkaline phosphatase (bone ALP; EC 3.1.3.1) in human sera, are described. Bone ALP is bound to streptavidin-coated microwells by a single biotinylated anti-bone ALP monoclonal antibody. Antigen is detected by the addition of p-nitrophenyl phosphate. The assay is performed at room temperature in &lt;90 min. Imprecision was 2.3–6.1% with a detection limit of 0.6 μg/L. Method comparison of bone ALP measurements with the Tandem-MP Ostase assay and the mass-based Tandem-R Ostase assay (n = 285) indicated regression statistics of Tandem-MP Ostase = 1.03 Tandem-R Ostase + 0.22 μg/L, Sy‖x = 4.0 μg/L, r = 0.97. Serum bone ALP values in apparently healthy men and in pre- and postmenopausal women were also similar between the two Ostase assay formats. Liver ALP reactivity determined using the slope and heat inactivation methods was similar in both Ostase assays. Liver ALP reactivity ranged from 3 μg/L (heat inactivation) to 6 μg/L (slope method) per 100 U/L of liver ALP activity, whereas bone ALP reactivity was 37 μg/L per 100 U/L of bone ALP activity, indicating a liver ALP relative reactivity of 8.1–16.2%. Similar results were obtained with the Alkphase-B bone ALP immunoassay. The Tandem-MP Ostase bone ALP assay demonstrated increased concentrations of serum bone ALP in conditions where bone metabolism is increased and showed a rapid, temporal decrease in serum bone ALP in Paget disease patients on bisphosphonate therapy. In conclusion, the Tandem-MP Ostase assay for serum bone ALP is a rapid, simple, robust nonisotopic alternative to the Tandem-R Ostase immunoradiometric assay that provides an accurate and sensitive assessment of bone turnover.
CO-Oximetry interference by perflubron emulsion: comparison of hemolyzing and nonhemolyzing instruments
Authors: Shepherd A P , Steinke J M 
Published: January 2020

Abstract
Perflubron emulsion is expected to be in clinical use soon as a non-hemoglobin blood substitute. A preliminary report indicates that this new oxygen-carrying fluorocarbon interferes with the measurements of CO-oximeters. Therefore, we have quantified the interference that perflubron causes in the measurements of eight widely used oximeters and CO-oximeters. The AVL Omni 6, CC270, IL482, IL682, and OSM3 are conventional CO-oximeters that hemolyze blood samples before analyzing them. In contrast, the AVOXimeters 1000 and 4000 and the IL Synthesis 35 make their measurements without hemolyzing the samples. Because perflubron is expected to be used most frequently on surgical patients in a hemodiluted state, we conducted all tests on human erythrocytes suspended in plasma at a hemoglobin concentration standardized to 70 g/L (7 g/dL) and with oxyhemoglobin saturation set at 97%. When perflubron was added to the blood samples, the nonhemolyzing CO-oximeters were not seriously affected by perflubron concentrations in and above the therapeutic range. In contrast, some of the hemolyzing CO-oximeters experienced concentration-dependent interference in their measurements of all analytes except total hemoglobin concentration. Thus, we conclude that the nonhemolyzing CO-oximeters provide an effective means for determining whether a hemolyzing CO-oximeter is experiencing clinically important interference in blood from patients receiving perflubron.
Direct reverse transcription-PCR on oligo(dT)-immobilized polypropylene microplates after capturing total mRNA from crude cell lysates
Authors: Hamaguchi Yohei , Aso Yoshimasa , Shimada Hiroshi , Mitsuhashi Masato 
Published: January 2020

Abstract
To simplify gene expression analysis, oligo(dT)-immobilized polypropylene microplates were used serially to capture mRNA, synthesize cDNA, and amplify specific genes. The amounts of immobilized oligonucleotide, hybridized mRNA, and synthesized cDNA were quantified fluorometrically using either Yoyo-1 or AttoPhos. The immobilized oligonucleotides captured ∼40–55% of mRNA directly from crude cell lysates. Hybridized mRNA was then amplified by one-step reverse transcription (RT)-PCR with rTth polymerase or two-step PCR with initial cDNA synthesis followed by PCR, where the latter exhibited more sensitivity. In two-step RT-PCR, microplates can be reused for multiple PCRs with the same or different primer sets because synthesized cDNA was covalently attached to the plates at its 5′ end. We believe this microplate may be acceptable as a platform for various mRNA expression analyses, including basic research, drug screening, and molecular toxicology, as well as for molecular pathological diagnostics.
Detection of intracellular antigens by flow cytometry: comparison of two chemical methods and microwave heating
Authors: Millard Isabelle , Degrave Etienne , Philippe Marianne , Gala Jean-Luc 
Published: January 2020

Abstract
Detection of intracellular antigens by flow cytometry requires effective fixation and permeabilization of the cell membrane. This study compares three fixation/permeabilization techniques: two commercial chemical reagents, the ORTHOPermeaFixTM (OPF) and the FIX&PERM Cell Permeabilization Kit® (F&P), and a novel method based on microwave heating (MWH). They have been applied to the detection of two nuclear (p53 and rb/p105) and two cytoplasmic (bcl-2 and mdr-1/gp-170) antigens, using positive- and negative-control cell lines and peripheral blood mononuclear cells. Western blotting was performed as a control of protein expression. For the four antigens assessed, cellular morphology, discrimination between intact cells and debris, percentage of positive cells, and mean fluorescence intensity were examined. For this last parameter, the assessment of the MWH technique was performed using SD and a graphical approach inspired by the concepts described by Bland and Altman (Lancet 1986;346:1085–7) as well as Petersen et al. (Clin Chem 1997;43:2039–46). The statistical analysis shows that MWH is comparable to the commercial methods and that its reproducibility is also equivalent to OPF and F&P. As assessed for some of the most clinically relevant intracytoplasmic and intranuclear antigens, the MWH method appears to be a valuable and inexpensive alternative. It is worth noting that, unlike commercial reagents, MWH altered surface antigens. Interestingly, this feature, which would prevent cell selection on the basis of combined membrane and intracellular epitopes, is associated with a decrease of nonspecific background fluorescence.
Clinical outcome and economic impact of aminoglycoside peak concentrations in febrile immunocompromised patients with hematologic malignancies
Authors: Binder Lutz , Schiel Xaver , Binder Claudia , Menke Cristina Fernandes Almeida , Schüttrumpf Silke , Armstrong Victor W , Unterhalt Michael , Erichsen Norbert , Hiddemann Wolfgang , Oellerich Michael 
Published: January 2020

Abstract
The aim of this study was to investigate the clinical and economic significance of aminoglycoside peak concentrations in febrile neutropenic patients with hematologic malignancies. Sixty-one patients were treated according to protocol II of the Paul-Ehrlich-Gesellschaft: initial application of gentamicin or tobramycin in combination with a cephalosporin or ureidopenicillin and, after 3 days, a potential change of antibiosis to be decided in case of nonresponse. At the same time, samples were collected by an independent controller. We found a significant dependence of clinical outcome on aminoglycoside peak concentrations (P = 0.004). Twelve of 17 patients with peak concentrations &gt;4.8 mg/L, but only 13 of 44 patients with concentrations ≤4.8 mg/L, responded to initial therapy. Average infection-related costs per patient with peak values &gt;4.8 mg/L were US$1429, $1790, and $1701 for nursing, diagnostics, and therapeutics, respectively (total $4920). Expenses for patients with peak concentrations ≤4.8 mg/L were ∼1.8-fold higher (average total $8718). If all 61 patients had achieved peaks &gt;4.8 mg/L, the potential savings would have totalled $167 112. We conclude that neutropenic patients form a target group for successful pharmacokinetic intervention and cost saving.
Development of new cancer chemoprevention agents: role of pharmacokinetic/pharmacodynamic and intermediate endpoint biomarker monitoring
Authors: Lieberman Ronald , Crowell James A , Hawk Ernest T , Boone Charles W , Sigman Caroline C , Kelloff Gary J 
Published: January 2020

Abstract
Recently, several promising strategies have been advanced for improving the efficiency of new agent development. These include pharmacokinetic/pharmacodynamic (PK/PD) and intermediate endpoint bio-marker (IEB) monitoring. Here, we review their essential role as practical tools for guiding the evaluation of agents for cancer chemoprevention (CP) and provide examples of CP agents that utilize these approaches. Several important categories of IEBs are delineated, including histologically based (intraepithelial neoplasias and nuclear morphometry). The use of select IEBs combined with a Bayesian method for clinical trial monitoring for rapid identification of ineffective or promising agents is discussed. The similarities between IEB and TDM are described. Finally, we present future tools for enhanced monitoring of CP agents that will impact on laboratory medicine and are also applicable to many other drug classes, e.g., laser capture microdissection and cDNA chip microarrays that assess gene expression patterns of precancerous and cancerous lesions.
Isolation and identification of a C39 demethylated metabolite of rapamycin from pig liver microsomes and evaluation of its immunosuppressive activity
Authors: Nickmilder Marc J M , Latinne Dominique , De Houx Jean-Paul , Verbeeck Roger K , Lhoëst Georges J J 
Published: January 2020

AbstractWe studied in vitro metabolism of rapamycin using pig liver microsomes. After extraction of the metabolites from the incubation medium, the crude metabolite extract was submitted to normal and subsequently to reversed-phase HPLC chromatography. We describe in the current study a metabolite of retention time 23.2 min collected from reversed-phase HPLC and identified by fast atom bombardment mass spectrometry (MS) and electrospray MS-MS as a C39 demethylated rapamycin metabolite. In vitro immunosuppressive activity of this metabolite, determined by the mixed lymphocyte reaction, was negligible compared with that of the parent compound. The decrease of in vitro immunosuppressive activity compared with the parent compound is likely to be attributed to important structural modifications of the rapamycin binding region to the FK-506 binding protein.
Strontium absorption and excretion in normocalciuric subjects: relation tocalcium metabolism
Authors: Vezzoli Giuseppe , Baragetti Ivano , Zerbi Simona , Caumo Andrea , Soldati Laura , Bellinzoni Piera , Centemero Antonella , Rubinacci Alessandro , Moro GianLuigi , Bianchi Giuseppe 
Published: January 2020

AbstractThe relationships of Sr intestinal absorption and renal excretion with biohumoral factors regulating Ca metabolism were studied in 47 normocalciuric subjects with Ca kidney stones. Sr concentrations were measured in serum and urine after an oral load of stable Sr (30.2 μmol/kg body wt). Enteral absorption of the ion (9.77 ± 0.438 mmol · L−1 · min, 240 min after Sr administration), expressed as the area under the plasma concentration–time curve (AUC), and renal clearance (CRE) in these subjects during the test (2.80 ± 0.336 mL/min) were not different from values for 27 controls. CRE was not correlated with AUCs. Plasma concentrations of parathyroid hormone (PTH) negatively correlated with AUCs (P &lt;0.01) and correlated with CRE after one outlier was excluded (P &lt;0.05). Plasma concentrations of 1,25-dihydroxyvitamin D correlated positively with AUCs (P &lt;0.01) when normalized to the plasma concentration of PTH. Multiple stepwise regression showed that PTH and phosphatemia were significantly related to AUC values at 240 min (P &lt;0.01). These findings suggest that Sr absorption and excretion reflect the regulation of Ca metabolism, but some differences in renal handling of the two ions may exist.