bluuebunny/crossref_metadata_2025_split huggingface dataset:
Glycohemoglobin: A Primary Predictor of the Development or Reversal of Complications of Diabetes Mellitus
Authors: Krishnamurti Uma , Steffes Michael W 
Published: January 2020

AbstractBackground: Diabetes mellitus is a major health problem worldwide with long-term micro- and macrovascular complications responsible for a majority of its morbidity and mortality. The development and progression of these complications relate strongly to glycemic control.Methods: We reviewed the literature extensively for studies that relate glycemic control to the development and progression of diabetic complications. We discuss the problems of standardizing glycohemoglobin measurements for monitoring diabetic therapy and also consider recently developed electrospray ionization mass spectrometry methods that have been considered as candidate reference methods for estimation of glycohemoglobin.Results: Several clinical trials and studies have clearly shown that improved glycemic control is strongly associated with decreased development and/or progression of complications in both type 1 and type 2 diabetes mellitus. Irrespective of the methods used for estimating glycohemoglobin, these results underline the importance of glycohemoglobin for guiding therapy of diabetes mellitus. Recently developed candidate reference methods promise to yield greatly improved standardization for the measurement of glycohemoglobin.Conclusions: Glycohemoglobin measurement remains the optimal indicator of glycemic control in diabetic patients, but translation of findings from clinical trials to clinical practice worldwide demands consistent values across all assays. To ensure that the important prognostic information still applies to all diabetic patients with the application of the reference method(s), the hemoglobin A1c values reported in the major clinical trials will have to be translated into statistically and computationally compatible values based on the new reference system(s).
In-Tube DNA Methylation Profiling by Fluorescence Melting Curve Analysis
Authors: Worm Jesper , Aggerholm Anni , Guldberg Per 
Published: January 2020

Abstract
Background: Most PCR assays for detection of 5-methylcytosine in genomic DNA entail a two-step procedure, comprising initial PCR amplification and subsequent product analysis in separate operations that usually require manual transfer. These methods generally provide information about methylation of only a few CpG dinucleotides within the target sequence.
Methods: An in-tube methylation assay is described that integrates amplification of bisulfite-treated DNA and melting analysis by using a thermal cycler coupled to a fluorometer (LightCycler). DNA melting curves were acquired by measuring the fluorescence of a double-stranded DNA-binding dye (SYBR Green I) during a linear temperature transition.
Results: Analysis of a region comprising 11 CpG sites at the SNRPN promoter CpG island showed that the melting temperature (Tm) differed by ∼3 °C between unmethylated and fully methylated alleles. This assay could easily distinguish patients with Prader-Willi syndrome or Angelman syndrome from individuals without these conditions. Melting curve analysis also allowed resolution of methylation “mosaicism” at the p15Ink4b promoter in bone marrow samples from patients with acute myeloid leukemia (AML). AML samples representing pools of heterogeneously methylated p15Ink4b alleles showed broadened melting peaks with overall Tms between those of the unmethylated and fully methylated alleles.
Conclusions: Integration of PCR and fluorescence melting analysis may be useful for simple and cost-effective detection of aberrant methylation patterns.
Discontinuation of the Bleeding Time Test without Detectable Adverse Clinical Impact
Authors: Lehman Christopher M , Blaylock Robert C , Alexander Donald P , Rodgers George M 
Published: January 2020

Abstract
Background: The bleeding time (BT) test predicts a higher bleeding complication rate in populations at risk for inherited or acquired platelet dysfunction, but it is of limited assistance in evaluating individual patients. There are no reports of clinical outcomes after discontinuation of the BT test.
Methods: Interviews with a subset of the physicians who had ordered the BT test before discontinuation of the test were conducted. The total number of platelet-aggregation tests, the mean number of monthly, unmodified platelet units transfused, the incidence of kidney biopsy complications, and the number of doses of 1-deamino-8-d-arginine vasopressin (DDAVP) administered 5 months before and after discontinuation of the BT test were compared. We recorded the rates of bleeding complications in the Major Surgery Risk Pool during the 12 months before and the 5 months after the discontinuation of the BT test.
Results: Clinicians reported they did not significantly change their preprocedural work-ups, postpone an invasive procedure, experience an increase in bleeding complications, or increase their use of blood products after discontinuation of the BT test. Platelet-aggregation tests (n = 9, before and after), platelet transfusions (P = 0.958), and DDAVP administration (before = 24; after = 10) did not increase after discontinuation of the BT test. The rate of postprocedural bleeding complications did not increase significantly in either Major Surgery Risk Pool cases (&lt;3ς deviation from the mean rate) or in patients undergoing renal biopsies (P = 0.225 for decrease in hematocrit; P = 1.000 for the percentage of patients transfused) after discontinuation of the BT test.
Conclusions: Our study failed to identify a clinically significant, negative impact of discontinuing the BT test.
Cystatin C Is an Independent Predictor of Fasting and Post-Methionine Load Total Homocysteine Concentrations among Stable Renal Transplant Recipients
Authors: Aras Ömer , Tsai Michael Y , Hanson Naomi Q , Bailey Robert , Rao Gundu , Hunninghake Donald B 
Published: January 2020

AbstractBackground: An increased prevalence of hyperhomocysteinemia with an increased incidence of cardiovascular disease events has been reported among stable renal transplant recipients (RTRs). Preliminary studies in a small number of these individuals have shown that serum creatinine and cystatin C, both markers of kidney function and glomerular filtration rate, are independent determinants of fasting tHcy concentrations; however, determinants of tHcy concentrations after a methionine load have not been studied.Methods: We determined the prevalence of both fasting and 4-h post-methionine load (PML) tHcy concentrations in 78 stable RTRs and compared the role of cystatin C with the role of serum creatinine as determinants of fasting and PML tHcy.Results: Of the 78 RTRs, 21 (26.9%) had fasting and PML tHcy within the respective reference intervals, and 57 (73.1%) had increased plasma tHcy. Of these 57 RTRs, 22 had fasting hyperhomocysteinemia, 9 had PML hyperhomocysteinemia, and 26 had combined hyperhomocysteinemia (both fasting and PML). Unadjusted Pearson correlations showed that fasting plasma tHcy correlated with both cystatin C (r = 0.564; P &lt;0.001) and creatinine (r = 0.519; P &lt;0.001) and that increases in PML tHcy modestly correlated with cystatin (r = 0.205; P = 0.072), but not creatinine (r = 0.057; P = 0.624). General linear regression modeling with stepwise analysis of covariance showed that both cystatin C (partial R = 0.554; P &lt;0.001) and creatinine (partial R = 0.535; P &lt;0.001) were independent predictors of fasting tHcy, but of the two, only cystatin C (partial R = 0.242; P = 0.035) was an independent predictor of increased PML tHcy.Conclusions: Clinically stable RTRs have an excess prevalence of moderate hyperhomocysteinemia, and additional cases can be detected by methionine loading. Both creatinine and cystatin C are independent predictors of fasting tHcy in these individuals; however, only cystatin C is a determinant of tHcy concentration after a methionine load, probably because cystatin C is a more sensitive marker of glomerular filtration rate than serum creatinine.
Supersensitive Time-resolved Immunofluorometric Assay of Free Prostate-specific Antigen with Nanoparticle Label Technology
Authors: Soukka Tero , Paukkunen Janika , Härmä Harri , Lönnberg Stefan , Lindroos Hanne , Lövgren Timo 
Published: January 2020

AbstractBackground: The extreme specific activity of the long-lifetime fluorescent europium(III) chelate nanoparticles and the enhanced monovalent binding affinity of multivalent nanoparticle-antibody bioconjugates are attractive for noncompetitive immunoassay.Methods: We used a noncompetitive, two-step immunoassay design to measure free prostate-specific antigen (PSA). Europium(III) chelate nanoparticles (107 nm in diameter) were coated with a monoclonal anti-PSA antibody (intrinsic affinity, 6 × 109 L/mol). The nanoparticle-antibody bioconjugates had an average of 214 active binding sites per particle and a monovalent binding affinity of 7 × 1010 L/mol. The assay was performed in a low-fluorescence microtitration well passively coated with an another monoclonal anti-PSA antibody (affinity, 2 × 1010 L/mol), and the europium(III) fluorescence was measured directly from the bottom of the well by a standard time-resolved microtitration plate fluorometer.Results: The detection limit (mean + 2 SD) was 0.040 ng/L (7.3 × 105 molecules/mL), and the dynamic detection range covered four orders of magnitude in a 3-h total assay time. The imprecision (CV) over the whole assay range was 2–10%. The detection limit of the assay was limited by the fractional nonspecific binding of the bioconjugate to the solid phase (0.05%), which was higher than the nonspecific binding of the original antibody (&lt;0.01%).Conclusions: The sensitivity of the new assay is equal to that of the ambient-analyte, microspot immunoassay and will be improved by use of optimized, high binding-site density nanoparticle-antibody bioconjugates with reduced nonspecific binding and improved monovalent binding affinity.
Screening for Mutations and Polymorphisms in the Genes KCNH2 and KCNE2 Encoding the Cardiac HERG/MiRP1 Ion Channel: Implications for Acquired and Congenital Long Q-T Syndrome
Authors: Larsen Lars Allan , Andersen Paal Skytt , Kanters Jørgen , Svendsen Ida Hastrup , Jacobsen Joes Ramsøe , Vuust Jens , Wettrell Göran , Tranebjærg Lisbeth , Bathen Jørn , Christiansen Michael 
Published: January 2020

AbstractBackground: The voltage-gated, rapid-delayed rectifier current (IKr) is important for repolarization of the heart, and mutations in the genes coding for the K+-ion channel conducting this current, i.e., KCNH2 for the α-subunit HERG and KCNE2 for the β-subunit MiRP1, cause acquired and congenital long Q-T syndrome (LQTS) and other cardiac arrhythmias.Methods: We developed a robust single-strand conformation polymorphism-heteroduplex screening analysis, with identical thermocycling conditions for all PCR reactions, covering all of the coding exons in KCNH2 and KCNE2. The method was used to screen 40 unrelated LQTS patients.Results: Eleven mutations, of which six were novel, were found in KCNH2. Interestingly, six mutations were found in the region of the gene coding for the Per-Arnt-Sim (PAS) and PAS-S1 regions of the HERG protein, stressing the need to examine the entire gene when screening for mutations. No mutations were found in KCNE2, suggesting that direct involvement of MiRP1 in LQTS is rare. Furthermore, four novel single-nucleotide polymorphisms (SNPs) and one amino acid polymorphism (R1047L) were identified in KCNH2, and one novel SNP and one previously known amino acid polymorphism (T8A) were found in KCNE2.Conclusions: The potential role of rare polymorphisms in the HERG/MiRP1 K+-channel should be clarified with respect to drug interactions and susceptibility to arrhythmia and sudden death.
Intermethod Discordance for α-Fetoprotein Measurements in Fanconi Anemia
Authors: Cassinat Bruno , Darsin Diane , Guardiola Philippe , Toubert Marie-Elisabeth , Rain Jean-Didier , Gluckman Eliane , Schlageter Marie-Hélène 
Published: January 2020

Abstract
Background: The significantly higher serum α-fetoprotein (AFP) in patients with Fanconi anemia (FA) than in non-FA aplastic patients has potential diagnostic utility, but the increase is method-dependent. The aim of this study was to compare five AFP assays on FA and non-FA samples and to investigate possible explanations for FA-specific discrepancies.
Methods: Two methods available in our laboratory (Kryptor and IMx) were compared on 59 FA and 27 non-FA patient samples. Kryptor, Immulite, Elecsys, Immuno-I, and Elsa-2 methods were then compared on 14 FA and 14 non-FA patient samples. The AFP glycosylation profile was analyzed by electrophoretic separation in a lectin-containing gel.
Results: With all six methods, AFP values were significantly higher in FA than in non-FA patients, but the diagnostic precision and optimal cutoff values varied. Indeed, two methods reached 100% sensitivity and specificity, but in other methods, one or both of these parameters were significantly &lt;100%. Neither heterophilic antibodies nor a specific glycosylation profile was detected in FA samples.
Conclusions: AFP results are method-dependent in FA. New methods must be evaluated before use in differential diagnosis of aplastic patients.
Excitatory Amino Acids in Cerebrospinal Fluid of Patients with Acute Head Injuries
Authors: Hong Zhang , Xinding Zhang , Tianlin Zhang , Liren Chen 
Published: January 2020

Abstract
Background: The excitatory amino acids (EAAs) glutamate (Glu) and aspartate (Asp) play a role in the pathogenesis of postischemic and posttraumatic brain insult. The changes of EAAs in cerebrospinal fluid (CSF) of patients with traumatic brain injury are incompletely understood.
Methods: We used reversed-phase HPLC with precolumn derivatization with o-phthalaldehyde and fluorescence detection to measure Glu and Asp in CSF of 42 patients with acute head injury and 9 control adults without neurologic diseases. We assessed the Glasgow Coma Scale (GCS) on admission, the main lesion patterns on computed tomography (CT) scan within 24 h post trauma, and the Glasgow Outcome Scale (GOS) 3 months post injury.
Results: The mean concentrations of Glu and Asp in CSF in the brain-injured group were significantly higher than those of the control group (Glu, P &lt;0.001; Asp, P &lt;0.05). In patients admitted within 24 h after severe injury (n = 13), peak Glu values appeared within 48 h in 11 patients (85%), and the mean value remained higher than control values at day 7 (P &lt;0.02). The concentrations of EAAs were higher in patients with severe injuries (GCS ≤8) than in those with milder injuries (Glu, P &lt;0.001; Asp, P &lt;0.05). GCS and peak EAAs correlated negatively (Glu, rs = −0.5706, P &lt;0.001; Asp, rs = −0.5503, P &lt;0.001). The patients with focal brain contusion on initial CT scan (n = 8) had a significantly lower peak Glu value than the patients with other patterns (n = 8–15; P &lt;0.02 to 0.001). The peak value of EAAs in the poor-outcome group (including severe disability, vegetative state, and death) was significantly higher than in the good-outcome group (good recovery and moderate disability; Glu, P &lt;0.001; Asp, P &lt;0.01); GOS was closely correlated to the EAA values (Glu, rs = 0.5737, P &lt;0.001; Asp, rs = 0.5470, P &lt;0.001).
Conclusions: The EAA concentrations in CSF increase after acute head injury and remain higher for at least 1 week post injury in severely injured patients. The more severe the trauma, the more obvious the excitotoxicity induced by EAAs and the worse the outcome.
Heterozygous M3Mmalton α1-Antitrypsin Deficiency Associated with End-Stage Liver Disease: Case Report and Review
Authors: Canva Valérie , Piotte Sandrine , Aubert Jean-Pierre , Porchet Nicole , Lecomte-Houcke Martine , Huet Guillemette , Zenjari Tahar , Roumilhac Didier , Pruvot François-René , Degand Pierre , Paris Jean-Claude , Balduyck Malika 
Published: January 2020

Abstractα1-Antitrypsin (α1AT) deficiency is an autosomal recessive disorder that can cause pulmonary emphysema and liver disease. We report here the case of a 59-year-old woman who was admitted to hospital for evaluation of jaundice. She had no history of hepatitis or childhood liver disease. She had never received a blood transfusion, nor had she abused drugs or alcohol. Transjugular liver biopsy was then performed and revealed a micronodular cirrhosis. Ten months later, because of persistent liver cell failure and ascites, she underwent an orthotopic liver transplantation. Investigation of α1AT system in the proband revealed a substantial decrease in serum α1AT associated with a low elastase inhibitory capacity. The Pi phenotype revealed a PiM-like profile. Sequencing of exons 1–5 demonstrated the presence of the M3 allele. Moreover, a triple nucleotide deletion was detected in exon 2 of one allele. This caused an “in-phase” frameshift, coding for a protein deficient in a single Phe residue, which corresponded to the Mmalton variant. After liver biopsy, periodic acid-Schiff-positive acidophilic bodies resistant to diastase digestion were observed in the cytoplasm of hepatocytes. These results demonstrated that our patient had a heterozygous M3Mmalton α1AT genotype related to a deficiency phenotype. This observation is the first of a patient with heterozygous Mmalton genotype associated with an α1AT deficiency that induced severe liver disease requiring orthotopic liver transplantation.
Molecular Detection of Mycobacterium tuberculosis: Impact on Patient Care
Authors: Kaul Karen L 
Published: January 2020

Abstract
Background: Nucleic acid amplification technologies such as PCR are revolutionizing the detection of infectious pathogens such as tuberculosis (TB). Amplification technology offers the potential for the diagnosis of TB in a few hours with a high degree of sensitivity and specificity. However, molecular assays neither replace nor reduce the need for conventional smear and culture, speciation, and antibiotic sensitivity assays.
Methods: We undertook prospective studies of sputum samples to assess the performance of two PCR-based assays for the detection of TB as well as the impact of more rapid availability of test results on patient care.
Results: The sensitivity of both the in-house and Amplicor PCR assays was 100% for smear-positive sputa. For smear-negative sputa (two sputum samples collected during the first 24 h of hospitalization), the sensitivity was 85% for our in-house PCR assay and 74% for the Roche PCR assay. Approximately 10% of the smear- and culture-negative sputa yielded positive PCR results; however, more than one-half of these were positive with both the in-house and Amplicor assays, suggesting the presence of TB DNA or organisms. Several of these came from patients whose other samples grew Mycobacterium tuberculosis during the same admission, and others came from patients who had previously treated TB. Overall, the specificities of the in-house and Amplicor PCR assays in smear-negative patients were 86% and 93%, respectively.
Conclusions: Molecular detection of slow-growing pathogens such as M. tuberculosis have the potential to improve clinical care through a dramatic reduction in the time required for detection and may provide substantial savings in the overall cost of care of a patient compared with conventional smear, culture, and speciation alone, despite the fact that conventional assays must still be performed for speciation of nontuberculous mycobacteria and for full assessment of antibiotic sensitivity.
Detection of Antinuclear Antibodies by Use of an Enzyme Immunoassay with Nuclear HEp-2 Cell Extract and Recombinant Antigens: Comparison with Immunofluorescence Assay in 307 Patients
Authors: Hayashi Nobuhide , Kawamoto Tomoko , Mukai Masahiko , Morinobu Akio , Koshiba Masahiro , Kondo Shinichi , Maekawa Soichiro , Kumagai Shunichi 
Published: January 2020

Abstract
Background: A new enzyme immunoassay (EIA) for automated detection of antinuclear antibodies (ANAs) uses a mixture of HEp-2 cell extracts and multiple recombinant nuclear antigens immobilized on beads. We compared this EIA and an immunofluorescence (IF) assay in a large group of patients and controls.
Methods: We studied 492 healthy individuals and 307 patients with connective tissue diseases (CTDs). Sera were tested by an automated EIA (COBAS® Core HEp2 ANA EIA; Roche Diagnostics) and IF. Samples were also tested for eight disease-specific antibodies, including antibodies against U1RNP, Sm, SSA/Ro, SSB/La, Scl-70, Jo-1, dsDNA, and centromere.
Results: Areas under ROC curves for the EIA were greater than (P = 0.008–0.012) or numerically identical to areas for the IF method for each of six CTDs studied. ROC areas for EIA were 0.98 (95% confidence interval, 0.95–0.99), 0.99 (0.96–1.00), and 0.99 (0.98–1.00) in systemic lupus erythematosus (n = 111), systemic sclerosis (n = 39), and mixed connective tissue disease (n = 33), respectively. For all 258 CTD patients with conditions other than rheumatoid arthritis (RA), the sensitivity and specificity of the IF method at a cutoff dilution of 1:40 were 92% and 65%, respectively, vs 93% and 79% for the EIA at a cutoff of 0.6. For the IF method at a cutoff dilution of 1:160, sensitivity and specificity were 81% and 87%, respectively, vs 84% and 94%, respectively, for the EIA at a cutoff of 0.9. For 207 sera containing at least one of eight disease-specific ANAs, positivities for the EIA and the IF method were 97.1% and 97.6%, respectively, at cutoffs of 0.6 and 1:40 (P = 0.76).
Conclusions: An EIA that can be performed by a fully automated instrument distinguishes CTDs (except RA) from healthy individuals with both higher sensitivity and specificity than the IF method when the cutoff index was set at 0.9. Moreover, it can be used to exclude the presence of disease-specific ANAs by setting the cutoff index at 0.6 with almost the same efficacy as the IF method.
Is the Thrombopoietin Assay Useful for Differential Diagnosis of Thrombocytopenia? Analysis of a Cohort of 160 Patients with Thrombocytopenia and Defined Platelet Life Span
Authors: Gouin-Thibault Isabelle , Cassinat Bruno , Chomienne Christine , Rain Jean-Didier , Najean Yves , Schlageter Marie-Hélène 
Published: January 2020

AbstractBackground: Thrombopoietin (TPO), the major hormone controlling platelet production, has been measured in thrombocytopenias with discordant results. The aim of our work was to assess the value of the TPO assay for differential diagnosis of thrombocytopenias in a large cohort of patients classified according to the results of their platelet isotopic study.Methods: We measured TPO (R&D Systems) in serum of 160 thrombocytopenic patients referred to our department for platelet life span isotopic studies. We classified patients as follows: (a) idiopathic or autoimmune thrombocytopenia group (ITP; patients with increased platelet destruction and shortened platelet life span; n = 67); (b) pure genetic thrombocytopenia group (patients with decreased platelet production, normal platelet life span, and without bone marrow aplasia; n = 55); (c) bone marrow aplasia group (BM; patients with decreased platelet production, normal platelet life span, and bone marrow aplasia; n = 13).Results: In patients with pure genetic thrombocytopenia, TPO (median, 55 ng/L) was not different from TPO in patients with ITP (median, 58 ng/L) or controls (n = 54; median, 51 ng/L). Only in patients with bone marrow aplasia was TPO significantly higher (median, 155 ng/L) and negatively correlated to the platelet count (r2 = 0.5014).Conclusions: Although the median serum TPO is increased in thrombocytopenia with decreased platelet production from bone marrow aplasia, it does not differentiate patients with pure genetic thrombocytopenia from those with ITP.