bluuebunny/crossref_metadata_2025_split huggingface dataset:
Changes in Bone Turnover in Patients with Anorexia Nervosa during Eleven Weeks of Inpatient Dietary Treatment
Authors: Heer Martina , Mika Claudia , Grzella Ina , Drummer Christian , Herpertz-Dahlmann Beate 
Published: January 2020

AbstractBackground: Many adolescents with anorexia nervosa suffer from severe osteopenia and osteoporosis. We hypothesized that individualized nutrition therapy may improve bone turnover in anorectic patients.Methods: We studied 19 female patients [mean age, 14.2 ± 1.4 years; mean body weight, 39.3 ± 5.4 kg; mean body mass index (BMI), 14.2 ± 1.4 kg/m2] with anorexia nervosa (International Classification of Diseases-10: F50.0, F50.1) for a period of 3 months. Nutrition therapy began at the end of the first week and included individualized hypercaloric diets, high calcium intake (2000 mg/day), and administration of vitamin D (400 IU/day). Blood samples were taken at baseline and again in weeks 3, 7, and 11. We measured serum calcium, parathyroid hormone, bone formation and resorption markers, insulin-like growth factor 1 (IGF-1), and leptin.Results: Mean BMI increased significantly, from 14.2 ± 1.4 to 17.1 ± 0.7 kg/m2 (P = 0.000001), during the course of treatment, whereas serum total calcium and phosphate concentrations remained unchanged. The bone formation markers procollagen-I carboxy-terminal propeptide and bone alkaline phosphatase almost doubled (P = 0.006). Both IGF-1 (P = 0.00001) and leptin (P = 0.000005) increased significantly by week 11. Parallel to this, the serum concentration of C-telopeptide, a bone resorption marker, decreased significantly (P = 0.009).Conclusions: Nutritional rehabilitation, possibly as a result of increasing IGF-1 and leptin concentrations, may increase bone formation. It therefore provides additional objective evidence of the importance of nutrition for bone.
5,10-Methylenetetrahydrofolate Reductase (MTHFR) Assay in the Forward Direction: Residual Activity in MTHFR Deficiency
Authors: Suormala Terttu , Gamse Gertraud , Fowler Brian 
Published: January 2020

AbstractBackground: Assay of methylenetetrahydrofolate reductase (MTHFR), a key enzyme in homocysteine metabolism, is important for the study of severe and mild deficiency states. Because the conventional assay measures in the reverse direction, lacks sensitivity, and uses nonphysiologic substrates, the exact measurement and characterization of residual activity in easily accessible tissues have been difficult.Methods: To measure MTHFR in the physiologic direction, we determined the NADPH-dependent conversion of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate by use of HPLC with fluorescence detection.Results: MTHFR activity in control fibroblast in the presence of FAD was maximal between pH 6.3 and 6.9, increased linearly up to 40 min and 80 μg protein/assay, and showed Kms of 30 μmol/L for NADPH and 26 μmol/L for 5,10-methylenetetrahydrofolate. Intraassay variation (CV) was 10%, interassay variation was 7.2%, and variation among 10 subcultures of the same cell line was 18%. Mean (SD) control activity was 431 (150) μU/mg protein (range, 242–910; n = 75), which is 2.5-fold higher than that with the reverse assay. After heat treatment (46 °C for 5 min), the activity showed a trimodal distribution corresponding to the 677TT (thermolabile; 15%), 677CT (35%), and 677CC (51%) genotypes. We found clearly measurable activity ranging from 2.6% to 25.6% of the mean control value in 15 patients with MTHFR deficiency, including 11 cell lines with zero activity in the reverse assay. Ten patients had complete enzyme deficiency.Conclusion: This assay allows reliable determination of residual activity in mutant fibroblasts and characterization of kinetic parameters for natural substrates.
Evaluation of the Anticoagulants EDTA and Citrate, Theophylline, Adenosine, and Dipyridamole (CTAD) for Assessing Platelet Activation on the ADVIA 120 Hematology System
Authors: Macey Marion , Azam Urooj , McCarthy Desmond , Webb Lee , Chapman E Sabrinah , Okrongly David , Zelmanovic David , Newland Adrian 
Published: January 2020

Abstract
Background: Monitoring of platelet activation by the ADVIA® 120 Hematology System requires an anticoagulant and protocol that ensures that platelets are sphered and their activation status is not altered artifactually in vitro.
Methods: Blood from healthy controls was collected into tripotassium EDTA; citrate, theophylline, adenosine, and dipyridamole (CTAD); or a combination of both (E/C) and stored at ambient temperature or at 4 °C (E/C only) and then analyzed between 0 and 180 min later on the ADVIA 120. In addition, immunofluorescent flow cytometry was used to identify activated platelets and platelet-leukocyte aggregates.
Results: In blood stored with all three anticoagulants, the platelet count changed little, but the mean platelet volume (MPV) at first decreased and then increased, whereas the mean platelet component (MPC; an indicator of activation) changed in a reciprocal manner. The changes in MPV and MPC, which reflect platelet sphering and swelling, were greatest between 30 and 60 min in blood stored at ambient temperature, irrespective of which anticoagulant was used, and between 60 and 180 min when blood anticoagulated with E/C was stored at 4 °C. In all anticoagulants, the percentages of platelets expressing CD62P and of leukocytes in platelet-leukocyte aggregates increased significantly (P &lt;0.01) over 180 min at ambient temperature. Only minimal (&lt;2%) increases occurred when blood with E/C was stored at 4 °C.
Conclusions: When determining platelet activation ex vivo on the ADVIA 120, blood should be collected into E/C, stored at 4 °C, and analyzed between 60 and 180 min later; these conditions ensure maximum platelet sphering without concurrent artifactual platelet activation.
Determination of α-Fetoprotein in Human Serum by a Quartz Crystal Microbalance-based Immunosensor
Authors: Chou Shu-Fen , Hsu Win-Lin , Hwang Jing-Min , Chen Chien-Yuan 
Published: January 2020

Abstract
Background: Increased α-fetoprotein (AFP) in adult plasma is considered an early indication of hepatocellular carcinoma and teratoblastoma. The aim of this study was to develop a rapid method to measure AFP in human serum by use of a direct immunosensor based on a quartz crystal microbalance (QCM).
Methods: A self-assembled monolayer prepared by the cystamine method was applied to immobilize anti-AFP monoclonal antibodies on the gold surface of a quartz crystal. The frequency shifts of the QCM were measured and related to AFP concentrations. The piazoimmunosensor used no labeled reagent and no pretreatment of samples.
Results: Ten cycles of measurements could be performed on the gold surface of the same crystal regenerated with a solution of glycine-HCl. A linear relationship existed between the frequency shifts (Hz) and the log values of AFP concentrations from 0.1 to 100 μg/L in buffer and human serum. When used for 15 days, the frequency shifts were all &gt;95% of those on the response at the first day. The regression equation was y = 1.03x − 0.06 (Sy|x = 3.92; r = 0.9987) for this QCM method and RIA in 29 clinical human serum samples.
Conclusions: The QCM sensor can measure AFP in buffer and human serum and offers advantages of high specificity, reusability, low detection limit, no label or sample pretreatment, and low sample requirement. The assay format of the immunosensor was more rapid and simpler than conventional methods.
Performance of a Revised Cardiac Troponin Method That Minimizes Interferences from Heterophilic Antibodies
Authors: Kim Wesley J , Laterza Omar F , Hock Karl G , Pierson-Perry James F , Kaminski Delores M , Mesguich Martine , Braconnier François , Zimmermann Régine , Zaninotto Martina , Plebani Mario , Hanna Atef , Cembrowski George S , Scott Mitchell G 
Published: January 2020

AbstractBackground: Recent guidelines for use of cardiac troponin to detect cardiac damage and for cardiovascular risk stratification have made increasingly sensitive troponin assays important. Troponin assays continue to be plagued by interferences caused by heterophilic antibodies (HAs). We evaluated the performance of a revised cardiac troponin I (cTnI) assay designed to have increased analytical sensitivity and to minimize the effect of HAs.Methods: The revised Dade Behring Dimension® cTnI assay was evaluated according to NCCLS EP5-A at five institutions. Plasma samples from 14 309 patients were assayed by the original Dimension cTnI assay. To identify samples that may have interfering HAs, samples with values &gt;1.4 μg/L were reanalyzed on the Dade Behring Stratus® CS cTnI assay. Samples with possible interfering antibodies were also analyzed before and after selective absorbance studies on the revised Dade Behring Dimension cTnI assay.Results: The limit of quantification in the revised method was 0.1 μg/L with imprecision (CV) of 11–17% at 0.1 μg/L. Values correlated well with the Stratus CS cTnI method: revised = 1.06(original) + 0.01; r = 0.98, Sy|x = 0.25 μg/L). Falsely increased results consistent with myocardial infarction by the original Dimension cTnI assay and presumably attributable to HAs were identified in 0.17% of all patients with samples submitted for cTnI analysis. The revised Dimension cTnI assay eliminated the interference in 17 of 25 samples identified and greatly decreased the interference in the other 8.Conclusions: The revised Dimension cTnI method greatly minimizes the effect of interfering HAs. It also exhibits analytical performance characteristics consistent with recent guidelines for use of this assay to detect cardiac damage.
Biochemical Markers and Hematologic Indices in the Diagnosis of Functional Iron Deficiency
Authors: Thomas Christian , Thomas Lothar 
Published: January 2020

AbstractBackground: The hypochromic red cell is a direct indicator of functional iron deficiency (ID) in contrast to the majority of biochemical markers, which measure functional ID indirectly via iron-deficient erythropoiesis. The aim of this study was to evaluate the extent to which these biochemical markers can distinguish ID from anemia of chronic disease (ACD) as well as from the combined state of functional ID/ACD, using red cell hemoglobinization as the gold standard.Methods: We studied 442 patients with various disease-specific anemias and 154 nonanemic patients. As indicators of red cell hemoglobinization, we measured the reticulocyte hemoglobin content (CHr) and the proportion of hypochromic red cells (HYPO), using an Advia 120 hematology analyzer. Ferritin, transferrin, transferrin saturation, and the concentration of the soluble transferrin receptor (sTfR) were determined by ELISA and immunoturbidimetric assay. The sTfR/log ferritin ratio (sTfR-F index) was used as an additional marker for biochemical identification of iron-deficient erythropoiesis.Results: In a control group (n = 71), the 2.5 percentile values were 28 pg for CHr and 5% for HYPO. These values were used to indicate unimpaired red cell hemoglobinization and absence of functional ID. In patients with deficient red cell hemoglobinization but no acute-phase response (APR), iron-deficient erythropoiesis was indicated by serum ferritin and sTfR-F index values ≤20.8 μg/L and &gt;1.5, respectively. Corresponding values in patients with APR were ≤61.7 μg/L and &gt;0.8, respectively. The positive likelihood ratios for the biochemical markers and the sTfR-F index for identifying iron-restricted erythropoiesis in patients with and without APR were 2.6–6.9 and 4.3–16.5, respectively.Conclusion: In APR patients, biochemical markers demonstrate weaknesses in the diagnosis of functional ID as defined by hematologic indices. Use of diagnostic plots to illustrate the relationship between the sTfR-F index and CHr allows the progression of ID to be identified, regardless of whether an APR is present.
Real-Time PCR Technology for Cancer Diagnostics
Authors: Bernard Philip S , Wittwer Carl T 
Published: January 2020

Abstract
Background: Advances in the biological sciences and technology are providing molecular targets for diagnosing and treating cancer. Current classifications in surgical pathology for staging malignancies are based primarily on anatomic features (e.g., tumor-node-metastasis) and histopathology (e.g., grade). Microarrays together with clustering algorithms are revealing a molecular diversity among cancers that promises to form a new taxonomy with prognostic and, more importantly, therapeutic significance. The challenge for pathology will be the development and implementation of these molecular classifications for routine clinical practice.
Approach: This article discusses the benefits, challenges, and possibilities for solid-tumor profiling in the clinical laboratory with an emphasis on DNA-based PCR techniques.
Content: Molecular markers can be used to provide accurate prognosis and to predict response, resistance, or toxicity to therapy. The diversity of genomic alterations involved in malignancy necessitates a variety of assays for complete tumor profiling. Some new molecular classifications of tumors are based on gene expression, requiring a paradigm shift in specimen processing to preserve the integrity of RNA for analysis. More stable markers (i.e., DNA and protein) are readily handled in the clinical laboratory. Quantitative real-time PCR can determine gene duplications or deletions. Furthermore, melting curve analysis immediately after PCR can identify small mutations, down to single base changes. These techniques are becoming easier and faster and can be multiplexed. Real-time PCR methods are a favorable option for the analysis of cancer markers.
Summary: There is a need to translate recent discoveries in oncology research into clinical practice. This requires objective, robust, and cost-effective molecular techniques for clinical trials and, eventually, routine use. Real-time PCR has attractive features for tumor profiling in the clinical laboratory.
Human Kallikrein Gene 5 (KLK5) Expression by Quantitative PCR: An Independent Indicator of Poor Prognosis in Breast Cancer
Authors: Yousef George M , Scorilas Andreas , Kyriakopoulou Lianna G , Rendl Laura , Diamandis Maria , Ponzone Riccardo , Biglia Nicoletta , Giai Maurizia , Roagna Riccardo , Sismondi Piero , Diamandis Eleftherios P 
Published: January 2020

AbstractBackground: KLK5 is a newly discovered human kallikrein gene. Many kallikrein genes have been found to be differentially expressed in various malignancies, and prostate-specific antigen (PSA; encoded by the KLK3 gene) is the best tumor marker for prostate cancer. Like the genes that encode PSA and other kallikreins, the KLK5 gene was found to be regulated by steroid hormones in the BT-474 breast cancer cell line.Methods: We studied KLK5 expression in 179 patients with different stages and grades of epithelial breast carcinoma by quantitative reverse transcription-PCR (RT-PCR), using LightCycler® technology. An optimal cutoff point equal to the detection limit (65th percentile) was used. KLK5 values were then compared with other established prognostic factors in terms of disease-free (DFS) and overall survival (OS).Results: High KLK5 expression was found more frequently in pre-/perimenopausal (P = 0.026), node-positive (P = 0.029), and estrogen receptor-negative (P = 0.038) patients. In univariate analysis, KLK5 overexpression was a significant predictor of reduced DFS (P &lt;0.001) and OS (P &lt;0.001). Cox multivariate analysis indicated that KLK5 was an independent prognostic factor for DFS and OS. KLK5 remained an independent prognostic variable in the subgroups of patients with large tumors (&gt;2 cm) and positive nodes. Hazard ratios derived from Cox analysis and related to DFS and OS were 2.48 (P = 0.005) and 2.37 (P = 0.009), respectively, for the node-positive group and 3.03 (P = 0.002) and 2.94 (P = 0.002), respectively, for patients with tumor sizes &gt;2 cm. KLK5 expression was also associated with statistically significantly shorter DFS (P = 0.006) and OS (P = 0.004) in the subgroup of patients with grade I and II tumors.Conclusions: KLK5 expression as assessed by quantitative RT-PCR is an independent and unfavorable prognostic marker for breast carcinoma.
Effect of Aspirin Treatment on Serum Concentrations of Lipoprotein(a) in Patients with Atherosclerotic Diseases
Authors: Akaike Masashi , Azuma Hiroyuki , Kagawa Ayako , Matsumoto Kazuya , Hayashi Ikuro , Tamura Katsuya , Nishiuchi Takeshi , Iuchi Takahiko , Takamori Nobuyuki , Aihara Ken-ichi , Yoshida Tomonori , Kanagawa Yasuhiko , Matsumoto Toshio 
Published: January 2020

AbstractBackground: Increased serum lipoprotein(a) [Lp(a)] is an independent risk factor for atherosclerosis. We previously reported that aspirin reduced Lp(a) production by cultured hepatocytes via the reduction of apolipoprotein(a) [apo(a)] gene transcription.Methods: We evaluated both the effect of aspirin treatment (81 mg/day) on serum Lp(a) concentrations and the correlation between the degree of reduction in serum Lp(a) and the type of apo(a) isoform in 70 patients with coronary artery disease or cerebral infarction.Results: Aspirin lowered serum Lp(a) concentrations to ∼80% of the baseline values in patients with high Lp(a) concentrations (&gt;300 mg/L). The percentage of decrease in serum Lp(a) was larger in patients with high Lp(a) than in patients with low Lp(a) (&lt;300 mg/L), irrespective of apo(a) isoform size. The decreases in serum Lp(a) in high Lp(a) patients with both the high-molecular-weight and the low-molecular-weight isoforms were positively correlated with the baseline Lp(a) concentrations.Conclusions: Because the secretory efficiencies of apo(a) in the same isoform are likely to be similar, the difference in serum Lp(a) concentrations in patients having the same apo(a) isoform depends on the transcriptional activity of the apo(a) gene. These findings suggest that aspirin decreases serum Lp(a) concentrations via a decrease in apo(a) gene transcription more effectively in patients with high transcriptional activity of this gene.
Limited Sampling Strategy for the Estimation of Mycophenolic Acid Area under the Curve in Adult Renal Transplant Patients Treated with Concomitant Tacrolimus
Authors: Pawinski Tomasz , Hale Mike , Korecka Magda , Fitzsimmons William E , Shaw Leslie M 
Published: January 2020

AbstractBackground: Significant relationships between the mycophenolic acid (MPA) area under the concentration–time curve (AUC0–12h) and the risks for acute rejection and side effects have been reported. We developed a practical method for estimation of MPA AUCs. Regression equations were developed using repeated cross-validation for randomly chosen subsets, characterized statistically, and verified for acceptable performance.Methods: Twenty-one renal transplant patients receiving 0.5 or 1.0 g of mycophenolate mofetil twice daily and concomitant tacrolimus provided a total of 50 pharmacokinetic profiles. MPA concentrations were measured by a validated HPLC method in 12 plasma samples collected at predose and at 30 and 60 min; 2, 3, 4, 6, 8, 9, 10, 11, and 12 h; 1 and 2 weeks; and 3 months after transplantation. Twenty-six 1-, 2-, or 3-sample estimation models were fit (r2 = 0.341–0.862) to a randomly selected subset of the profiles using linear regression and were used to estimate AUC0–12h for the profiles not included in the regression fit, comparing those estimates with the corresponding AUC0–12h values, calculated with the linear trapezoidal rule, including all 12 timed MPA concentrations. The 3-sample models were constrained to include no samples past 2 h.Results: The model using c0h, c0.5h, and c2h was superior to all other models tested (r2 = 0.862), minimizing prediction error for the AUC0–12h values not included in the fit (i.e., the cross-validation error). The regression equation for AUC estimation that gave the best performance for this model was: 7.75 + 6.49c0h + 0.76c0.5h + 2.43c2h. When we applied this model to the full data set, 41 of the 50 (82%) estimated AUC values were within 15% of the value of AUC0–12h calculated using all 12 concentrations.Conclusions: This limited sampling strategy provides an effective approach for estimation of the full MPA AUC0–12h in renal transplant patients receiving concomitant tacrolimus therapy.
Riboflavin, Flavin Mononucleotide, and Flavin Adenine Dinucleotide in Human Plasma and Erythrocytes at Baseline and after Low-Dose Riboflavin Supplementation
Authors: Hustad Steinar , McKinley Michelle C , McNulty Helene , Schneede Jørn , Strain JJ , Scott John M , Ueland Per Magne 
Published: January 2020

AbstractBackground: Vitamin B2 exists in blood as riboflavin and its cofactors, flavin mononucleotide (FMN) and FAD. The erythrocyte glutathione reductase activation coefficient (EGRAC) has traditionally been used to assess vitamin B2 status in humans. We investigated the relationships of EGRAC and plasma and erythrocyte concentrations of riboflavin, FMN, and FAD in elderly volunteers and their responses to riboflavin administration.Methods: EGRAC and plasma and erythrocyte concentrations of riboflavin, FMN, and FAD were determined in 124 healthy individuals with a mean age of 69 years. The same measurements were made in a subgroup of 46 individuals with EGRAC ≥1.20 who participated in a randomized double-blind 12-week intervention study and received riboflavin (1.6 mg/day; n = 23) or placebo (n = 23).Results: Median plasma concentrations were 10.5 nmol/L for riboflavin, 6.6 nmol/L for FMN, and 74 nmol/L for FAD. In erythrocytes, there were only trace amounts of riboflavin, whereas median FMN and FAD concentrations were 44 and 469 nmol/L, respectively. Erythrocyte FMN and FAD correlated with each other and with EGRAC and plasma riboflavin (P &lt;0.05). All variables except plasma FAD responded significantly to riboflavin supplementation compared with placebo (P ≤0.04). The strongest increases were for riboflavin in plasma (83%) and for FMN in erythrocytes (87%).Conclusions: Concentrations of all B2 vitamers except plasma FAD are potential indicators of vitamin B2 status, and plasma riboflavin and erythrocyte FMN may be useful for the assessment of vitamin B2 status in population studies.
Finite-size effects in exponential random graphs
Authors: Gorsky A , Valba O 
Published: January 2020

Abstract
In this article, we show numerically the strong finite-size effects in exponential random graphs. Particularly, for the two-star model above the critical value of the chemical potential for triplets a ground state is a star-like graph with the finite set of hubs at network density $p&lt;0.5$ or as the single cluster at $p&gt;0.5$. We find that there exists the critical value of number of nodes $N^{*}(p)$ when the ground state undergoes clear-cut crossover. At $N&gt;N^{*}(p),$ the network flows via a cluster evaporation to the state involving the small star in the Erdős–Rényi environment. The similar evaporation of the cluster takes place at $N&gt;N^{*}(p)$ in the Strauss model. We suggest that the entropic trap mechanism is relevant for microscopic mechanism behind the crossover regime.