bluuebunny/crossref_metadata_2025_split huggingface dataset:
Quantitative Analysis of the Bidirectional Fetomaternal Transfer of Nucleated Cells and Plasma DNA
Authors: Lo Y M Dennis , Lau Tze K , Chan Lisa Y S , Leung Tse N , Chang Allan M Z 
Published: January 2020

Abstract
Background: Recently, much interest has been generated on the fetomaternal transfer of nucleated cells and plasma DNA. However, there has been no systematic quantitative comparison of these two directions and two modalities of trafficking within the same study population.
Methods: The fetus-to-mother transfer of nucleated cells and plasma DNA in pregnant women carrying male babies was studied using a real-time quantitative PCR assay for the SRY gene. For mother-to-fetus transfer, real-time quantitative PCR assays for the insertion/deletion polymorphisms involving the glutathione S-transferase M1 and angiotensin-converting enzyme genes were used.
Results: Of the 50 informative mother-baby pairs, maternal DNA was detected in the cellular fraction of umbilical cord blood in 24% of cases (12 of 50), at a median fractional concentration of 2.6 × 10−4 (interquartile range, 1.7 × 10−4 to 3.6 × 10−4). In the plasma fraction of cord blood, maternal DNA was detected in 30% (15 of 50) of cases at a median fractional concentration of 3 × 10−3 (interquartile range, 1 × 10−3 to 1.6 × 10−2). For the other direction of trafficking, fetus-to-mother transfer of nucleated cells was detected in 26% of cases (13 of 50) at a median fractional concentration of 3.2 × 10−4 (interquartile range, 0.6 × 10−4 to 7.6 × 10−4). In the plasma fraction, fetal DNA was detected in 100% of maternal plasma (50 of 50) at a median fractional concentration of 3 × 10−2 (interquartile range, 1.4 × 10−2 to 5.3 × 10−2).
Conclusions: This study indicated that significantly more fetal DNA is present in the plasma of pregnant women compared with DNA from the cellular fraction of maternal blood. In addition, maternal DNA was demonstrated in both the cellular and plasma fractions of cord blood after delivery. This study has therefore determined the fundamental quantitative values for the bidirectional fetomaternal cellular and plasma DNA traffic.
Assessment of Mycophenolic Acid-induced Immunosuppression: A New Approach
Authors: Millan Olga , Oppenheimer Federic , Brunet Merce , Vilardell Jordi , Rojo Isabel , Vives Jordi , Martorell Jaume 
Published: January 2020

AbstractBackground: Mycophenolic acid (MPA), a metabolite of mycophenolate mofetil (MMF), is an immunosuppressive agent that inhibits inosine monophosphate dehydrogenase (IMPDH), a key enzyme in the ex novo synthesis of GTP. We measured IMPDH activity in peripheral blood mononuclear cells (PBMCs) from MMF-treated patients to evaluate the efficacy of MMF in individual patients.Methods: IMPDH activity was measured by 3H released from [2,8-3H]IMP that had been formed in the cells from added [2,8-3H]hypoxanthine in PBMCs of 35 renal transplant recipients treated with cyclosporin A and corticoids plus MMF: 2 g (n = 10), 1.5 g (n = 7), 1 g (n = 10), or 0 g (n = 8) per day. An alternative method, based on the capacity of the patients’ sera to inhibit spontaneous proliferation of the CEM cell line, was also analyzed.Results: The IMPDH activity of PBMCs in transplanted patients was highly variable. For the method based on CEM cell line proliferation: (a) cell proliferation was inhibited only in MMF-treated patients; (b) there was a clear postdose increase in inhibition; (c) inhibition was not affected by other immunosuppressants in vitro or in vivo; (d) inhibition from predose to predose sample was correlated; and (e) when the MMF dosage was &lt;20 mg · kg−1 · day−1, two groups of patients were identified, one that maintained a high inhibitory capacity in all dose intervals, and one with periods of low inhibitory capacity.Conclusions: Measurement of the inhibition of CEM cell line proliferation by sera from MMF-treated patients may be useful for evaluating the relative efficacy of MMF treatment in individual patients, especially those receiving low doses of MMF.
Stability of Common Analytes in Urine Refrigerated for 24 h before Automated Analysis by Test Strips
Authors: Froom Paul , Bieganiec Barbara , Ehrenrich Zahava , Barak Mira 
Published: January 2020

Abstract
Background: Central outpatient laboratories might find processing large numbers of urinary samples that arrive in the late afternoon inconvenient and refrigerate them overnight before testing. Furthermore, in certain settings clinics might have difficulty assuring that the urine arrives at the laboratory during the same day as the collection. Because the stability of urine samples for delayed automated dipstick analysis (Supertron) is unknown, after defining precision, we retested urines refrigerated for 24 h to determine stability.
Methods: Urinalysis was done twice on the same day and repeated after the sample was refrigerated for 24 h. Combur-10S (Roche Diagnostics) dipsticks were read automatically by a Supertron analyzer. Repeat tests on the same day were compared with tests after storage.
Results: Leukocyte esterase had high precision, but after storage ∼25% of the positive samples were less reactive (P &lt;0.005). Precision of hemoglobin retests was also high but declined significantly after storage for 24 h. Urine protein values increased after storage. The precision and stability were excellent for nitrites, glucose, and ketones.
Conclusions: The stability of the automated dipstick urinalysis varies with the substance tested. After refrigeration for 24 h, there is a risk of false-positive results for protein, false-negative results for leukocytes and erythrocytes, and little effect on glucose, nitrite, and ketone values.
Screening for Single-Nucleotide Polymorphisms Using Branch Migration Inhibition in PCR-amplified DNA
Authors: Lishanski Alla 
Published: January 2020

Abstract
Background: New methods are required for the exploration of the human genome by discovering sequence variations. This study evaluated the performance of a new method for screening a large number of samples for several DNA polymorphisms.
Methods: We used a homogeneous method based on inhibition of spontaneous branch migration by any sequence difference between two molecules of PCR-amplified DNA. A set of four PCR primers is required: a forward primer, either biotinylated or labeled with digoxigenin, and two reverse primers that share a priming domain but have different “tail” sequences at their 5′ ends. After PCR amplification, denaturation and reannealing of the single DNA strands produce doubly labeled cruciform structures, which dissociate by strand exchange. The presence of two different alleles in a sample causes complete inhibition of dissociation, and the association of biotin and digoxigenin is homogeneously detected using luminescent oxygen channeling immunoassay.
Results: The 90 samples of the Human Variation Panel (Coriell Cell Repositories) were screened for nine known single-nucleotide polymorphisms (SNPs) and one 5-bp deletion. The average signal-to-background ratio varied from ∼10 to 20. The frequency of the predominant allele for different SNPs varied from 51% to 88% overall. For some SNPs, it varied among the nine ethnic groups, e.g., 25–85% (average, 51%) for one SNP. The average heterozygosity varied from 0.17 to 0.54 and as much as 0.2–0.9 (average, 0.54) for one of the SNPs.
Conclusion: The method allows simple and rapid screening of a large number of samples for the presence of multiple alleles.
Carbohydrate-deficient Transferrin as a Marker of Chronic Alcohol Abuse: A Critical Review of Preanalysis, Analysis, and Interpretation
Authors: Arndt Torsten 
Published: January 2020

AbstractBackground: Carbohydrate-deficient transferrin (CDT) is used for diagnosis of chronic alcohol abuse. Some 200–300 reports on CDT have been published in impact factor-listed journals. The aims of this review were to condense the current knowledge and to resolve remaining issues on CDT.Approach: The literature (1976–2000) was searched using MEDLINE and Knowledge Server with “alcohol and CDT” as the search items. The data were reviewed systematically, checked for redundancy, and organized in sequence based on the steps involved in CDT analysis.Content: The review is divided into sections based on microheterogeneity of human serum transferrin (Tf), definition of CDT, structure of human serum CDT, pathomechanisms of ethanol-induced CDT increase, preanalysis, analysis, and medical interpretation (postanalysis). Test-specific cutoff values for serum CDT and causes of false positives and negatives for chronic alcohol abuse are discussed and summarized.Summary: Asialo- and disialo-Fe2-Tf, which lack one or two complete N-glycans, and monosialo-Fe2-Tf (structure remains unclear) are collectively referred to as CDT. Diminished mRNA concentration and glycoprotein glycosyltransferase activities involved in Tf N-glycan synthesis and increased sialidase activity most likely account for alcohol-induced increases in CDT. Knowledge about in vivo and in vitro effects on serum CDT is poor. Reliable CDT and non-CDT fractionation is needed for CDT measurement. Analysis methods with different analytical specificities and recoveries decreased the comparability of values and statistical parameters of the diagnostic efficiency of CDT. CDT is the most specific marker of chronic alcohol abuse to date. Efforts should concentrate on the pathomechanisms (in vivo), preanalysis, and standardization of CDT analysis.
Successful Diagnosis of Fetal Gender Using Conventional PCR Analysis of Maternal Serum
Authors: Honda Hiroshi , Miharu Norio , Ohashi Yoko , Ohama Koso 
Published: January 2020

Abstract
Background: Fetal DNA has been found in maternal plasma and serum. Diagnosis of fetal gender using maternal plasma and serum has been attempted in an effort to develop a new noninvasive method of prenatal diagnosis.
Methods: Peripheral blood samples were obtained from 61 pregnant women at 10–17 weeks of gestation before amniocentesis. DNA was extracted from 800 μL of each plasma or serum sample. To detect the Y-chromosome-specific sequences DYS14 and DYZ3 in the maternal plasma and serum, 40 cycles of PCR were carried out for each DNA extract. The PCR products were analyzed by 2.5% agarose gel electrophoresis and ethidium bromide staining, and the results were compared with the results of the cytogenetic analyses of amniocentesis.
Results: Cytogenetic analysis of amniocentesis revealed that 31 pregnant women had a male fetus and the remaining 30 pregnant women had a female fetus. Both DYS14 and DYZ3 were detected in 27 of the 31 plasma samples obtained from pregnant women carrying a male fetus and in all of 31 serum samples obtained from the same women. Neither DYS14 nor DYZ3 was detected in either the plasma or serum samples obtained from any of the 30 pregnant women carrying a female fetus.
Conclusion: PCR analysis of maternal serum can be used to diagnose fetal gender.
Tartrate-resistant Acid Phosphatase Isoform 5b as Serum Marker for Osteoclastic Activity
Authors: Janckila Anthony J , Takahashi Karen , Sun Susan Z , Yam Lung T 
Published: January 2020

AbstractBackground: Tartrate-resistant acid phosphatase (AcP) 5b is a marker of osteoclastic activity and bone resorption. Immunoassays for serum TRAcP may lack sensitivity and specificity because of the presence of non-bone isoform 5a. The purpose of this study was to isolate the serum isoforms, quantify their disease-related expressions, and test an improved immunoassay for TRAcP 5b.Methods: We separated TRAcP isoforms chromatographically from pooled sera of healthy, rheumatoid arthritis (RA) and endstage renal disease (ESRD) subjects. TRAcP isoforms were identified by electrophoresis and quantified by biochemical and immunochemical assays. Serum TRAcP activity in healthy, RA, and ESRD cohorts was assessed at pH 5.5 and 6.1, and compared with bone alkaline phosphatase (BAP) and N-telopeptides of type I collagen (NTx).Results: TRAcP isoforms 5a and 5b were present in all sera; 5b was identical to osteoclastic TRAcP. In serum from healthy subjects, 5a accounted for 87% of the enzyme protein but only 55% of the activity. In RA, both isoforms were increased two- to threefold in protein, but their specific activities were subnormal. In ESRD, only 5b was abnormal, being increased fivefold in protein and threefold in activity. In RA sera, TRAcP activity did not correlate with either BAP or NTx. In ESRD sera, TRAcP activity correlated with BAP and NTx only when measured at pH 6.1.Conclusions: All sera contained both TRAcP isoforms 5a and 5b, but only 5b was present in bone. TRAcP isoform expression was variable in different diseases. Measurement of TRAcP activity at pH 6.1 improves the specificity of immunoassay for isoform 5b.
Evidence for a Substantial Genetic Influence on Biochemical Liver Function Tests: Results from a Population-based Danish Twin Study
Authors: Bathum Lise , Petersen Hans Christian , Rosholm Jens-Ulrik , Hyltoft Petersen Per , Vaupel James , Christensen Kaare 
Published: January 2020

Abstract
Background: Biochemical liver function tests are widely used in the clinic and are some of the most frequently used tests in screening for diseases in older age groups. The aim of the present study was to estimate the relative importance of genetic and environmental factors to variations in these tests among the elderly.
Methods: We conducted a survey among Danish twins, 73–102 years of age, identified in the population-based Danish Twin Registry. Among the 2749 individuals in the study population, an interview was conducted with 79%. From these, a blood sample was collected from 290 same-sex twin pairs, total of 580 subjects, within a 6-month period and analyzed for alanine aminotransferase (ALT), lactate dehydrogenase (LDH), γ-glutamyltransferase (GGT), bilirubin, and albumin. The interview included questions about alcohol consumption and body mass index (BMI; self-calculated height and weight). Heritability (proportion of the population variance attributable to genetic variation) was estimated using structural-equation analyses before and after correction for alcohol consumption and BMI.
Results: Structural-equation analyses revealed a substantial heritability (35–61%) for the four biochemical liver function tests: ALT, GGT, LDH, and bilirubin. The remaining variation could be attributed to individuals’ nonfamilial environments. Adjustment for alcohol consumption and BMI had no influence on the heritability for ALT, GGT, LDH, and bilirubin. For albumin, two models fit equally well before adjustment for alcohol and BMI: a model including additive genetic and nonshared environmental factors (AE), and a model including shared and nonshared environmental factors (CE). After adjustment, the model including shared and nonshared environment was clearly the best fitting model.
Conclusions: For both males and females, the effect of genetic factors on the biochemical liver function tests ALT, GGT, LDH, and bilirubin is substantial and accounts for one-third to two-thirds of the variation among individuals 73–102 years of age. The heritability is equal for males and females and does not change notably after controlling for alcohol consumption and BMI. For albumin, no major impact of genetic factors was found independent of BMI and alcohol consumption. An understanding of the genetic mechanisms underlying biochemical liver function tests among the very old may be of value in the interpretation of these tests in this age group.
Correlation of Mycophenolic Acid Pharmacokinetic Parameters with Side Effects in Kidney Transplant Patients Treated with Mycophenolate Mofetil
Authors: Mourad Michel , Malaise Jacques , Chaib Eddour Djamila , De Meyer Martine , König Josiane , Schepers Raf , Squifflet Jean Paul , Wallemacq Pierre 
Published: January 2020

AbstractBackground: Mycophenolate mofetil (MMF) is widely used in organ transplantation to prevent acute rejection. Because MMF can produce hematologic and/or gastrointestinal toxicity, therapeutic monitoring is becoming mandatory. This study was designed to investigate the relationship between the clinical events and the pharmacokinetics of mycophenolic acid (MPA) in adult renal transplantation.Methods: Thirty-one adult kidney recipients were prospectively included in the study. MPA pharmacokinetic profiles (blood sampling at 0, 0.5, 1, 2, 4, 6, and 12 h after MMF oral dose) were obtained after transplantation (desired creatinine clearance, 40 mL/min), at 3 months after grafting, and at every clinical event (e.g., side effect or rejection). All patients received a 10-day course of anti-thymocyte globulin, cyclosporine, MMF (1 g twice daily), and steroids.Results: We divided the 31 patients into two groups (groups 1 and 2). Ten patients (32%; group 1) had uneventful outcomes, and 21 patients (68%; group 2) presented with MPA-related side effects. For groups 1 and 2, the MPA trough concentrations (Cmin) were 1.63 ± 1.07 and 2.29 ± 1.16 mg/L, respectively (P = 0.06), and the areas under the curve (AUCs) for MPA from t0 to t12 h (MPA-AUC0–12h) were 39.80 ± 15.29 and 62.10 ± 21.07 mg · h/L, respectively (P = 0.0005, two-sample t-test). Three patients experienced acute graft rejection after the oral MMF dose was reduced because of side effects. In this group, the MPA-Cmin and MPA-AUC were significantly lower by the time acute rejection occurred (1.00 ± 0.45 mg/L and 25.00 ± 6.20 mg · h/L, respectively). At a fixed dose (1 g twice per day), we compared the pharmacokinetic parameters of MPA [Cmin, the MPA concentration 30 min after the oral dose of MMF (C30), and AUC] according to the presence or absence of side effects in the two groups. Cmin and AUC did not differ between the two groups [Cmin = 2.22 ± 1.13 vs 2.17 ± 1.13 mg/L (P = 0.9); AUC = 66.82 ± 29.87 vs 55.70 ± 11.74 mg · h/L (P = 0.11)]; and C30 was significantly higher in group 2 than in group 1 (C30 = 32.99 ± 12.59 vs 7.45 ± 5.40 mg/L; P &lt;0.0001).Conclusions: Our results demonstrate a pharmacokinetic/pharmacodynamic relationship between MPA and clinical events. At a fixed dose of 2 g/day, a high C30 is associated with increased risk for side effects. This study suggests that dividing the MMF daily oral dose into more than two divided doses might prevent early MPA toxicity.
Measurement of Homocysteine and Other Aminothiols in Plasma: Advantages of Using Tris(2-carboxyethyl)phosphine as Reductant Compared with Tri-n-butylphosphine
Authors: Krijt Jakub , Vacková Martina , Kožich Viktor 
Published: January 2020

Abstract
Background: Aminothiols have been implicated in the pathogenesis of arteriosclerosis, and reliable methods are needed to determine their concentrations in body fluids. We present a comparison of two analytical methods and focus on the reduction of low-molecular weight and protein-mixed disulfides of homocysteine, cysteine, cysteinyl-glycine, and glutathione.
Methods: The plasma total aminothiol profile was determined by HPLC with fluorescence detection after derivatization with ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate. Disulfides and protein-bound aminothiols were reduced by either tri-n-butylphosphine (the TBP method) or tris(2-carboxyethyl)phosphine (the TCEP method); the effects of temperature, time of reduction, and concentration of reductants were evaluated.
Results: The intraassay imprecision (CV) was &lt;3% for all aminothiols using both methods. The interassay CVs for total cysteine (tCys), total cysteinyl-glycine (tCys-Gly), and total homocysteine (tHcy) were &lt;4% and &lt;8% for the TCEP and TBP methods, respectively, whereas for total glutathione (tGSH) the interassay CV was &gt;12% for both methods. Deming regression and Bland–Altman difference plots showed positive biases for total aminothiol concentrations determined by the TCEP method relative to the TBP method. The mean proportional biases were 65%, 27%, 6%, and 60% for tCys, tCys-Gly, tHcy, and tGSH, respectively. The calculated concentrations of total aminothiols by the TCEP method were less influenced by changes in temperature and concentration of reducing agent or by calibrator matrix.
Conclusions: The agreement between the TCEP and TBP methods was considerably lower for the determination of tCys, tCys-Gly, and tGSH than for tHcy. For total-aminothiol determination, the TCEP method yields better reproducibility and is more robust than the TBP method.
Proteomics in Early Detection of Cancer
Authors: Srinivas Pothur R , Srivastava Sudhir , Hanash Sam , Wright George L 
Published: January 2020

Abstract
Early detection is critical in cancer control and prevention. Biomarkers help in this process by providing valuable information about a the status of a cell at any given point in time. As a cell transforms from nondiseased to neoplastic, distinct changes occur that could be potentially detected through the identification of the appropriate biomarkers. Biomarker research has benefited from advances in technology such as proteomics. We discuss here ongoing research in this field, focusing on proteomic technologies. The advances in two-dimensional electrophoresis and mass spectrometry are discussed in light of their contribution to biomarker research. Chip-based techniques, such as surface-enhanced laser desorption, and ionization and emerging methods, such as tissue and antibody arrays, are also discussed. The development of bioinformatic tools that have and are being developed in parallel to proteomics is also addressed. This report brings into focus the efforts of the Early Detection Research Network at the National Cancer Institute in harnessing scientific expertise from leading institutions to identify and validate biomarkers for early detection and risk assessment.
Divergence between LDL Oxidative Susceptibility and Urinary F2-Isoprostanes as Measures of Oxidative Stress in Type 2 Diabetes
Authors: Devaraj Sridevi , Hirany Shaina V , Burk Raymond F , Jialal Ishwarlal 
Published: January 2020

AbstractBackground: Oxidative stress is pivotal in atherogenesis. Although the most widely used indirect assay to quantify oxidative stress is LDL oxidative susceptibility, direct assays such as urinary F2-isoprostanes have shown great promise.Methods: We evaluated the utility of both a direct measure of oxidative stress (urinary F2-isoprostanes) and an indirect measure of copper-catalyzed, LDL oxidation in a model of increased oxidative stress (diabetes). We also evaluated an enzyme immunoassay (EIA) method for urinary F2-isoprostanes with a gas chromatography–mass spectrometry method.Results: Excellent intraassay and interassay CVs of &lt;4% were obtained with our EIA method. A good correlation was obtained between the two methods (r = 0.80; n = 68) of F2-isoprostane measurement. An excellent correlation for F2-isoprostane concentrations was obtained between a timed collection vs 24-h urine (r = 0.96; n = 46). Baseline F2-isoprostane concentrations by EIA were significantly higher in both type 2 diabetics with and without macrovascular complications compared with controls (P &lt;0.001). Supplementation with α-tocopherol led to a significant reduction in F2-isoprostane concentrations in all diabetic patients compared with baseline values (2.51 ± 1.76 compared with 1.69 ± 1.32 ng/mg creatinine; P &lt;0.001). There were no significant differences in LDL oxidation in both diabetic groups compared with controls. α-Tocopherol supplementation led to significant increases in the lag phase of oxidation as measured by 3 indices in all groups.Conclusions: The measurement of urinary F2-isoprostanes provides a direct measure of in vivo lipid peroxidation and oxidative stress and appears to be superior to an indirect measure, e.g., LDL oxidative susceptibility, in type 2 diabetes.