bluuebunny/crossref_metadata_2025_split huggingface dataset:
Total testing process applied to therapeutic drug monitoring: impact on patients’ outcomes and economics
Authors: Schumacher Gerald E , Barr Judith T 
Published: January 2020

Abstract
The Total Testing Process (TTP) refers to the sequence of 11 steps of laboratory testing, beginning with a clinical question prompted by the patient–clinician encounter and concluding with the impact of the test result on patient care. TTP when applied to therapeutic drug monitoring (TDM) emphasizes that TDM must be considered a process involving a series of steps and interrelated activities and not viewed simply as a numerical value for a serum drug concentration. TTP is also an ideal format for organizing and identifying the system-related and patient-centered variables used in outcomes assessment of TDM, as well as providing a template for collecting the cost data needed for economic analyses. Examples are provided for improving application of TDM by practitioners, clinical laboratories, and educators.
Analytical goals in therapeutic drug monitoring
Authors: Bowers Larry D 
Published: January 2020

Abstract
The practice of therapeutic drug monitoring is based on several disciplines, including pharmacokinetics, pharmacodynamics, and chemical analysis. The impact of the analysis on the determination of pharmacokinetic parameters is not well appreciated. Analytical goals in therapeutic drug monitoring should be established by determining the nature of the problem to be solved, selecting the appropriate matrix and methodology to solve the problem, and developing valid analytical schemes that are performed competently with appropriate quality and interpreted within the framework of the problem.
Strategies for physician education in therapeutic drug monitoring
Authors: Bates David W , Soldin Steven J , Rainey Petrie M , Micelli Joseph N 
Published: January 2020

Abstract
Although therapeutic drug monitoring (TDM) is probably very useful overall, studies suggest that it could be used better. Many drug concentrations appear to have inappropriate indications or suboptimal timing, particularly in the inpatient setting. Undermonitoring is also a concern. Thus, it may be possible to both improve the quality of TDM and reduce the overall costs of care. Here we review approaches for improving the use of TDM and present some illustrative experiences. Specific approaches discussed include use of traditional approaches such as lectures and newsletters, multidisciplinary quality improvement efforts, formal TDM services, and use of the computer as a tool for education and behavior change. Computerized methods appear to hold substantial potential, particularly as more organizations develop better information systems, but other approaches are also effective and are complementary. To be most successful, interventions should consider all stages of the process.
Pharmacodynamic monitoring of immunosuppressive drugs
Authors: Yatscoff Randall W , Aspeslet Launa J , Gallant Heather L 
Published: January 2020

Abstract
Pharmacodynamic (PD) monitoring measures the biological response to a drug, which alone—or coupled with pharmacokinetics— provides a novel method for optimization of drug dosing. PD monitoring has been investigated by us and other investigators primarily for four immunosuppressive drugs: cyclosporine (CsA), azathioprine (AZA), mycophenolate mofetil (MMF), and rapamycin (RAPA). PD monitoring of CsA and MMF involves measuring the activity of the enzymes calcineurin and inosine monophosphate dehydrogenase, respectively. The PD of AZA is assessed by measuring the activity of thiopurine methyltransferase, which is induced by a metabolite of AZA, 6-mercaptopurine. The PD for RAPA involves measuring the activity of a P70 S6 kinase in lymphocytes. To date, the most detailed studies have been performed with PD monitoring of CsA and MMF. Similarities exist in the PD responses to CsA and MMF in renal-transplant patients. At trough concentrations in blood, both drugs reduce the activity of their target enzymes by only 50%; however, considerable interpatient variability is evident. Throughout the dosing interval, the enzyme activities parallel the respective drug concentrations. AZA treatment of renal-transplant patients who exhibited an increase in thiopurine methyltransferase activity from time of transplantation resulted in fewer episodes of active rejection. Additional clinical trials are currently underway to relate various pharmacokinetics and PD parameters to clinical response, to ascertain which provides the best guide for dosing. PD monitoring may provide an alternative approach to additional measurements of drug concentrations.
HPLC determination of ketamine, norketamine, and dehydronorketamine in plasma with a high-purity reversed-phase sorbent
Authors: Bolze Sébastien , Boulieu Roselyne 
Published: January 2020

Abstract
We developed an isocratic, selective, and very sensitive HPLC method for the determination of ketamine and its two main metabolites in plasma. The compounds were extracted from plasma by a liquid–liquid extraction with a dichloromethane:ethyl acetate mixture followed by an acidic back-extraction. Separation was achieved on a new stationary phase, Purospher RP-18 end-capped, with a mobile phase containing acetonitrile:0.03 mol/L phosphate buffer (23:77 by vol) adjusted to pH 7.2. Because of the high column efficiency and the significant improvement of peak symmetry, the quantification limit could be down to 5 μg/L for ketamine and norketamine (NK). The intraday and interday CVs ranged from 1.7% to 5.8% and 3.1% to 10.2% for all compounds respectively. The method is sensitive enough for monitoring ketamine, NK, and dehydroketamine in plasma during pharmacokinetic studies after an intravenous bolus of a low dose of ketamine.
Adrenomedullin(1–52) measured in human plasma by radioimmunoassay: plasma concentration, adsorption, and storage
Authors: Lewis Lynley K , Smith Mary W , Yandle Timothy G , Richards A Mark , Nicholls M Gary 
Published: January 2020

Abstract
We describe a specific and sensitive RIA for human adrenomedullin (AM)(1–52). The detection limit and the concentration required for 50% inhibition of binding were 0.1 and 1.2 fmol/tube, respectively. Cross-reactivities with AM(1–12), AM(13–52), calcitonin gene-related peptide, amylin, and other vasoactive hormones were negligible. AM immunoreactivity in normal subjects ranged from 2.7 to 10.1 pmol/L (n = 44). We investigated factors influencing the recovery and measurement of AM in the assay. Recovery of labeled AM (&gt;80%) was markedly higher than that of unlabeled AM (56%). Immunoreactivity of exogenous AM added to plasma decreased up to 70% over four freeze–thaw cycles, whereas endogenous AM was stable. Alkali-treated casein (1 g/L) reduced adsorption of AM to surfaces and significantly increased assay precision compared with bovine serum albumin (P &lt;0.0001). HPLC separation of extracted plasma verified the presence of AM(1–52). We suggest that considerable care is needed to ensure that accurate and reproducible results are obtained from studies quantifying this peptide.
Microchip systems for immunoassay: an integrated immunoreactor with electrophoretic separation for serum theophylline determination
Authors: Chiem Nghia H , Harrison D Jed 
Published: January 2020

Abstract
A glass microchip is described in which reagents and serum samples for competitive immunoassay of serum theophylline can be mixed, reacted, separated, and analyzed. The device functions as an automated microfluidic immunoassay system, creating a lab-on-a-chip. Electroosmotic pumping was used to control first the mixing of 50-fold-diluted serum sample with labeled theophylline tracer in a 1:1 ratio, followed by 1:1 mixing and reaction with anti-theophylline antibody. The 51-nL on-chip mixer gave the same concentration as dilution performed off-chip, within 3%. A 100-pL plug of the reacted solution was then injected into an electrophoresis separation channel integrated within the same chip. Measurements of free and bound tracer by fluorescence detection gave linear calibration curves of signal vs log[theophylline] between 0 and 40 mg/L, with a slope of 0.52 ± 0.03 and an intercept of −0.04 ± 0.04 after a 90-s reaction time. A detection limit of 0.26 mg/L in serum (expressed before the dilution step, actual concentration of 1.3 μg/L at the detector) was obtained. Recovery values were 107% ± 8% for 15 mg/L serum samples.
A competitive reverse transcription–PCR to study apolipoprotein ε gene expression
Authors: Rexin Martin , Feussner Giso 
Published: January 2020

AbstractWe developed a rapid and simple competitive reverse transcription-polymerase chain reaction for the quantification of apoε mRNA in human monocyte-derived macrophages. The method was applied, and its reliability was shown in patients with the familial lipoprotein disorder, type III hyperlipoproteinemia. Type III hyperlipoproteinemic patients express markedly higher concentrations of apoε mRNA when compared with healthy controls. Patients with this disease are usually (&gt;90%) homozygous for a receptor binding-defective isoform of apolipoprotein apo E (apo E2). The higher expression of apoε mRNA in the patients could, therefore, be a physiological mechanism to compensate for functionally defective apo E. The developed procedure might be valuable in assessment of apoε gene expression in human disease.
Direct determination of urinary iodine by inductively coupled plasma mass spectrometry using isotope dilution with iodine-129
Authors: Haldimann Max , Zimmerli Bernhard , Als Claudine , Gerber Hans 
Published: January 2020

AbstractAn inductively coupled mass spectrometric method was developed for the direct determination of iodine in urine. The application of isotope dilution analysis with added 129I offers new possibilities for automatic and accurate determinations. The sample preparation consists of dilution with an ammonia solution containing 129I. The validation was made by comparison with the results obtained in another laboratory by a spectrophotometric method based on the Sandell–Kolthoff reaction. Different regression models, including maximum likelihood estimation, were used to compare the methods. None of the models revealed analytical bias between the two methods. The urine samples analyzed for validation were from three persons previously exposed to an iodine bath and covered a concentration range of 0.2 to 2.8 μmol/L. A detection limit of 0.02 μmol/L, a within-run CV of 2.5%, and a between-run CV of 11.9% were estimated for the proposed method.
Serum kynurenine-to-tryptophan ratio increases with progressive disease in HIV-infected patients
Authors: Huengsberg Mia , Winer John B , Gompels Mark , Round Rachel , Ross Jonathan , Shahmanesh Mohsen 
Published: January 2020

Abstract
An alternative pathway of Trp metabolism involves the conversion of Trp to kynurenine by indoleamine-2,3-dioxygenase, which leads to synthesis of the neurotoxin, quinolinic acid. This study explores the relationship of indoleamine-2,3-dioxygenase activity with stages of HIV infection. Sera from 206 HIV-positive and 72 seronegative subjects were analyzed for Trp and kynurenine. The kynurenine-to-Trp (KT) ratio was calculated. The mean KT ratio of seronegative controls was 36.6 ± 10.9, and the median ratio was 34.9. The upper limit of the seronegative KT ratio, defined as mean + 2 SD, was 58.4. Patients with HIV infection showed a reciprocal relationship between the KT ratio, the CD4 count, and the stage of the disease. The median KT ratios for asymptomatic and AIDS patients were 50.5 and 117.0, respectively. This study shows that the serum Trp concentration is markedly decreased and that the kynurenine concentration is increased with immune stimulation in HIV infection. This may lead to changes in quinolinic acid and explain some of the pathogenesis of AIDS dementia.
Performance of Deming regression analysis in case of misspecified analytical error ratio in method comparison studies
Authors: Linnet Kristian 
Published: January 2020

Abstract
Application of Deming regression analysis to interpret method comparison data presupposes specification of the squared analytical error ratio (λ), but in cases involving only single measurements by each method, this ratio may be unknown and is often assigned a default value of one. On the basis of simulations, this practice was evaluated in situations with real error ratios deviating from one. Comparisons of two electrolyte methods and two glucose methods were simulated. In the first case, misspecification of λ produced a bias that amounted to two-thirds of the maximum bias of the ordinary least-squares regression method. Standard errors and the results of hypothesis-testing also became misleading. In the second situation, a misspecified error ratio resulted only in a negligible bias. Thus, given a short range of values in relation to the measurement errors, it is important that λ is correctly estimated either from duplicate sets of measurements or, in the case of single measurement sets, specified from quality-control data. However, even with a misspecified error ratio, Deming regression analysis is likely to perform better than least-squares regression analysis.
Antioxidant enzymes and fatty acid status in erythrocytes of Down syndrome patients
Authors: Pastor M-Cruz , Sierra Cristina , Doladé María , Navarro Elisabet , Brandi Nuria , Cabré Eduard , Mira Aurea , Serés Agustí 
Published: January 2020

AbstractThe excess of genetic information in patients with Down syndrome (DS) produces an increase in the catalytic activity of superoxide dismutase (SOD1), an antioxidant enzyme coded on chromosome 21. It has been suggested that an increase in oxidative stress in DS patients may cause adverse effects in the cell membranes through the oxidation of polyunsaturated fatty acids (PUFAs). The aim of this study was to evaluate the cellular antioxidant system by determining the catalytic activity of the SOD1, glutathione peroxidase (GPx), catalase (CAT), and glutathione reductase (GR) enzymes and the concentrations of α-tocopherol in red blood cells (RBCs) in a group of 72 DS patients. The profile of fatty acids in the phospholipids of RBC membranes was also evaluated. The activity of the erythrocyte antioxidant enzymes is significantly higher in the DS group than in the control group (SOD1, 635 ± 70 U/g Hb vs 476 ± 67 U/g Hb; CAT, 1843 ± 250 U/g Hb vs 1482 ± 250 U/g Hb; GPx, 23.2 ± 5.3 U/g Hb vs 21.5 ± 3.6 U/g Hb; and GR, 9.32 ± 1.4 U/g Hb vs 6.9 ± 1.3 U/g Hb, respectively). No differences were observed in RBC α-tocopherol concentrations between the two groups studied. Long-chain n6 PUFA (C20:3n6, C20:4n6) concentrations were increased in DS patients, suggesting enhanced Δ-6-desaturase activity. The long-chain n3 PUFA (docosahexenoic acid) does not appear to be affected by increased oxidative stress, probably because of the existence of compensatory antioxidant mechanisms.
Urinary measurement of transforming growth factor-β and type IV collagen as new markers of renal injury: application in diabetic nephropathy
Authors: Ellis Demetrius , Forrest Kimberly Y-Z , Erbey John , Orchard Trevor J 
Published: January 2020

Abstract
Urinary samples were concentrated rapidly and efficiently and were used to develop several protein assays that may be of value in monitoring individuals with progressive renal disorders. Transforming growth factor-β1 (TGF-β1) and retinol binding protein (RBP) were measured with modification of commercially available methods used to assay serum specimens; type 3 collagen (T3C) was measured with a new immunonephelometric assay. The precision characteristics of these assays are comparable with those reported for microalbuminuria. The clinical utility of measuring a panel of these markers was demonstrated in urine samples from 16 control subjects and from 46 individuals with insulin-dependent diabetes mellitus (IDDM) with various albumin excretion rates (AERs). TGF-β1 and T3C were used as markers of cytokine expression and of the renal fibrogenic process, whereas RBP excretion served as a marker of tubular injury or dysfunction. Compared with controls, T3C excretion was significantly increased in 18 normoalbuminuric and further increased in 13 microalbuminuric (AER 20 ≤ 200 μg/min) IDDM subjects. RBP excretion was increased in macroalbuminuric IDDM subjects (AER &gt;200 μg/min, overt nephropathy). Significant correlations were also found between AER and RBP in all but macroalbuminuric individuals, whereas TGF-β1 correlated with T3C excretion in controls and in normoalbuminuric diabetic subjects. Urinary RBP but not AER was an excellent predictor of diabetic nephropathy as defined by serum creatinine (P = 0.0001). This underscores the importance of an early tubulopathy in the subsequent development of glomerulopathy and overt nephropathy. The data suggest that longitudinal monitoring of a panel of urinary markers such as that used in the current study may better define their relevance in progressive glomerulosclerosis and may also provide greater insight into the mechanisms underlying such process.
4-Chlorotestosterone acetate metabolites in cattle after intramuscular and oral administrations
Authors: Le Bizec Bruno , Montrade Marie-Pierre , Monteau Fabrice , Gaudin Isabelle , Andre François 
Published: January 2020

Abstract
The use of 4-chlorotestosterone acetate by farmers for cattle fattening was recently demonstrated although the use of this anabolic steroid is strictly forbidden in the European Union. We investigated the metabolism of 4-chlorotestosterone acetate in the bovine species after intramuscular and oral administration. Nineteen metabolites were detected in urine after intramuscular injection, and eight metabolites were identified. For this purpose, preparative HPLC, mass spectrometry with different ionization modes (electronic impact and chemical ionization), and different acquisition techniques were used (high resolution, selected ion monitoring, and scan measurement). Metabolite stereoisomerism was determined on the basis of retention time and organic synthesis. 4-Chloroepitestosterone (M2), 4-chloroandrost-4-en-3α-ol-17-one (M3), and 4-chloroandrost-4-ene-3,17-dione (M4) were identified as the main urinary markers of intramuscular administration. On the other hand, 4-chloroandrost-4-ene-3α,17β-diol (M7), 4-chloroandrostan-3β-ol-17-one (M5), and M2 were the primary indicators of an oral administration. In addition, we have shown that 95% of the metabolites were sulfo-conjugated, except for M3, which was partially conjugated to glucuronic acid. Finally, the main metabolites (M2, M3, and M4) were easily identified for 1.5 months after intramuscular administration.
Indirect enzyme-linked immunosorbent assay for the quantitative estimation of lysergic acid diethylamide in urine
Authors: Kerrigan Sarah , Brooks Donald E 
Published: January 2020

AbstractA new antibody to lysergic acid diethylamide (LSD) was used to develop a novel indirect ELISA for the quantification of drug in urine. Evaluation of the new assay with the commercially available LSD ELISA (STC Diagnostics) shows improved performance. The test requires 50 μL of urine, which is used to measure concentrations of drug in the μg/L to ng/L range. The limit of detection was 8 ng/L compared with 85 ng/L in the commercial assay, and analytical recoveries were 98–106%. Our test detected 0.1 μg/L of LSD in urine with an intraassay CV of 2.4% (n = 8) compared with 6.0% for a 0.5 μg/L sample in the commercial assay (n = 20). The upper and lower limits of quantification were estimated to be 7 μg/L and 50 ng/L, respectively. Specificity was evaluated by measuring the extent of cross-reactivity with 24 related substances. Drug determination using the new assay offers both improved sensitivity and precision compared with existing methods, thus facilitating the preliminary quantitative estimation of LSD in urine at lower concentrations with a greater degree of certainty.
Paraquat-induced renal injury studied by 1H nuclear magnetic resonance spectroscopy of urine
Authors: Bairaktari Eleni , Katopodis Kostas , Siamopoulos Kostas C , Tsolas Orestes 
Published: January 2020

Abstract
The herbicide paraquat (1,1′-dimethyl-4,4′-bipyridylium dichloride; PQ), is a poison known to cause delayed mortality due to lung and kidney injuries. High-resolution proton nuclear magnetic resonance (1H NMR) spectroscopy has been extensively applied in evaluating nephrotoxicity by the characteristic perturbations in the excretion pattern of low molecular weight endogenous metabolites. The application of the method allows the rapid localization of the renal injury noninvasively. In this study, we report 1H NMR and conventional clinical chemistry urinalysis in two patients suffering from paraquat intoxication after overdose with suicidal intent. The alterations in the urine NMR spectrum suggest necrosis of the pars recta of the proximal renal tubules. The molecule of paraquat is also clearly detected in the same spectrum. In conclusion, the rapid screening of urine by NMR spectroscopy provides information about both the identity of the poison and the abnormal pattern of endogenous metabolites that characterize the location of the injury in renal tubules and reveals alterations in unusual metabolites that are not commonly measured.
Optimized PCR labeling in mutational and microsatellite analysis
Authors: Esposito Diana Liberata , Palmirotta Raffaele , Verı̀ Maria Concetta , Mammarella Sandra , D’Amico Franca , Curia Maria Cristina , Aceto Gitana , Crognale Stefania , Creati Beatrice , Mariani-Costantini Renato , Battista Pasquale , Cama Alessandro 
Published: January 2020

AbstractTo optimize the labeling and visualization of PCR products we tested different variables, including deoxynucleotide concentration and ratio, dilution of labeled product, number of PCR cycles, and use of one-step or nested labeling protocols. Labeling was achieved using a fixed amount of labeled dATP, whose relative specific activity was varied by adding increasing amounts of cold dATP. Optimal PCR-labeling intensity was reached at dATP concentrations between 0.9 and 7.0 μmol/L, with a peak at 1.8 μmol/L. This concentration corresponded to an optimal ratio between the increase in specific activity and the decrease in DNA yield. Nucleotide imbalances &gt;1:2 were not advantageous. Mutational analysis by single-strand conformational polymorphism (SSCP) was used to validate PCR-labeling protocols. The limiting nucleotide concentrations did not affect SSCP. Clear SSCP patterns were obtained using DNA templates of different sizes derived from several genes. SSCP patterns obtained using one-step or nested PCR-labeling protocols were equivalent and were visualized after overnight exposure, using [α35S]dATP as the label. Dilutions of labeled products ranging between 1:10 and 1:2.5 influenced SSCP patterns, and the lowest dilution tested produced better-defined and more-intense signals. Optimized SSCP conditions allowed the detection of novel and previously characterized nucleotide variants. Clear microsatellite typing was also obtained using optimized protocols and [α35S]dATP as the label.