bluuebunny/crossref_metadata_2025_split huggingface dataset:
Fast, manual, nonradioactive method for DNA sequencing
Authors: Debuire B , Chabli A , Frenoy N 
Published: January 2020

Abstract
We describe a protocol that allows nonradioactive detection of sequencing products after manual, direct, solid-phase sequencing of polymerase chain reaction-amplified DNA. The amplified DNA fragment to be studied is biotinylated at the 5' end of one of the two oligonucleotide primers used for amplification, allowing coupling to streptavidin-coated magnetic beads. The immobilized double-stranded DNA is then separated into single strands by alkaline treatment. A 5'-biotinylated sequencing primer is used after saturating with a biotin solution any possible remaining affinity sites on the streptavidin-coated magnetic beads. Sequencing is performed by using T7 DNA polymerase, and the sequencing products are electrophoresed in denaturing polyacrylamide sequencing gel. After transfer of the products to a nylon membrane, the sequencing pattern is revealed by chemiluminescence. Biotinylated alkaline phosphatase is bound to the 5' end of the sequencing primer via a streptavidin bridge and catalyzes the reaction by cleaving a phosphate group from a chemiluminescent substrate. The emitted photons are detected by exposing the membrane to x-ray film. This method is simple, rapid, and consistently successful and reproducible.
A method to quantify deviations from assay linearity
Authors: Krouwer J S , Schlain B 
Published: January 2020

Abstract
We present a statistical method to quantify deviations from linearity for assays that veer from linear assay responses. Our procedure handles the common case of unequally spaced analyte levels and nonconstant variance and provides a least-squares estimate with a confidence interval for the amount of deviation from assay linearity at a specified analyte concentration. This estimate of assay bias due to nonlinearity goes beyond the NCCLS EP6 lack-of-fit test, which tests for only the presence of nonlinearity. Knowing that nonlinearity is present is insufficient; users need to know the magnitude of the bias caused by nonlinearity. Our method can also be used with multifactor designs that estimate other systematic assay effects such as drift and carryover, thus obviating the need for a separate protocol to assess linearity. The procedure is carried out by adding extra columns to the design matrix corresponding to the concentration level(s) of interest. The extra columns, which replace the quadratic column, are orthogonal to all other columns. We describe a general method of constructing the new columns, and illustrate the procedure with a manual ammonia assay example dataset from EP6.
Primary aldosteronism in a patient with an aldosterone-producing adenoma
Authors: Pekarske S L , Herold D A 
Published: January 2020

Abstract
We describe an unusual patient presenting with a history of refractory hypertension and hypokalemia. Initial screening tests for adrenal hypertension were consistent with primary aldosteronism and an abdominal computed tomography scan showed an 8-mm left adrenal mass. However, adrenal venous sampling revealed markedly suppressed plasma aldosterone in the left adrenal vein but increased plasma aldosterone in the right adrenal vein. Therefore, on the basis of the clinical, radiographic, and laboratory findings, we concluded that the patient had a nonfunctioning adrenocortical adenoma on the left and an aldosterone-producing adrenocortical adenoma on the right, with the aldosteronoma resulting in hypertension and hypokalemia. Right adrenalectomy decreased the hypertension and corrected the hypokalemia. The right adrenal contained a 7-mm nodule microscopically consistent with the diagnosis of a cortical adenoma. The case highlights key steps and potential pitfalls in the evaluation of adrenal hypertension.
Development of third-generation immunochemiluminometric assays of follitropin and lutropin and clinical application in determining pediatric reference ranges
Authors: Pandian M R , Odell W D , Carlton E , Fisher D A 
Published: January 2020

Abstract
We developed dioxatane-based immunochemiluminometric assays (ICMAs) for lutropin (LH) and follitropin (FSH), using monoclonal antibodies. These ICMAs have a minimal detectable dose (analytical sensitivity) of 0.01 IU/L, extending the lower limit of sensitivity 10-fold (from 0.10 IU/L) when compared with immunoradiometric assays (IRMA) (second generation), and thus provide a true third-generation assay. Daytime FSH and LH concentrations were measured in 236 boys and 195 girls. Unlike the previous assays, all the samples had detectable concentrations of LH and FSH. In agreement with results from earlier methods, the present results indicate that for both sexes mean FSH and LH concentrations are relatively high during the early months of life, fall to baseline prepubertal concentrations by 12-18 months, and remain low until the onset of puberty. During puberty, the mean concentrations of FSH and LH increase significantly in both girls and boys with each stage of puberty, but there is considerable overlap between stages. These third-generation FSH and LH ICMAs reliably separate daytime plasma FSH and LH concentrations of prepubertal children from those of sexually mature children, and therefore can more reliably distinguish between the major causes of precocious puberty (e.g., gonadotropin dependent and independent). Our LH assay is also useful in monitoring the gonadotropin-releasing hormone therapy of patients with gonadotropin-dependent precocious puberty.
New reagent for colorimetric assay of calcium in serum
Authors: Chapoteau E , Czech B P , Zazulak W , Kumar A 
Published: January 2020

Abstract
We report a new single-reagent colorimetric assay for the determination of calcium in serum. The assay is based on a novel chromogenic tetracarboxylic acid chelating agent. The method has several advantages over the o-cresolphthalein complexone (CPC) method, including improved linearity, better accuracy, increased stability, and lower working pH (9.0). The calibration curve for the assay is linear up to 4 mmol/L. The results for 47 patients' samples correlated well with results obtained with an Instrumentation Laboratory atomic absorption spectrophotometer. The mean (+/- SD) calcium values for the new method were 2.27 +/- 0.06 mmol/L and 2.26 +/- 0.05 mmol/L for the comparison method. The on-system stability of the reagents is 1 week vs 8 h for the CPC method.
Microaffinity chromatographic separation and characterization of lipoprotein fractions in rat and mongolian gerbil serum
Authors: Tschantz J C , Sunahara G I 
Published: January 2020

Abstract
A rapid, routine microaffinity chromatographic procedure with spectrometric detection was evaluated to measure the concentration of total and free cholesterol, triglycerides, and phospholipids linked to the alpha- and beta-/pre-beta-lipoproteins in very small serum samples from small experimental animals and from humans. This procedure overcomes many of the limitations presented by methods involving ultracentrifugation, electrophoresis, or precipitation. Using this method, we report that in 35 adult male Mongolian gerbils (Meriones unguiculatus) fed a basal commercial diet with no cholesterol supplementation, the mean (+/- SD) concentration of total and free cholesterol linked to the alpha-lipoprotein fraction (in mmol/L) is 2.85 +/- 0.81 and 0.33 +/- 0.14, respectively; phospholipids were 1.99 +/- 0.41, and triglycerides were 0.64 +/- 0.42. The concentration (mmol/L) of total and free cholesterol associated with the beta-/pre-beta-lipoprotein fraction is 0.92 +/- 0.26 and 0.22 +/- 0.06, respectively; phospholipids were 0.28 +/- 0.17, and triglycerides were 0.49 +/- 0.31.
Automated closed-vessel system for in vitro diagnostics based on polymerase chain reaction
Authors: Findlay J B , Atwood S M , Bergmeyer L , Chemelli J , Christy K , Cummins T , Donish W , Ekeze T , Falvo J , Patterson D 
Published: January 2020

Abstract
An automated system for polymerase chain reaction (PCR) amplification and detection combats false-positive results caused by "PCR product carryover." The system uses a single vessel for both PCR amplification and the subsequent detection of PCR products, eliminating the need to handle PCR products in an open environment and risk product carryover. The sample and PCR reagents are introduced into one compartment within the vessel, and amplification occurs as they are thermally cycled. Other compartments contain the reagents for detection of PCR products. Pressure from a roller provides for sequential delivery of the contents of the compartments to a detection area. The PCR products are biotinylated at their 5' ends during amplification through the use of biotinylated primers. After delivery to the detection area, they are specifically captured by hybridization with immobilized oligonucleotide probes. Subsequent reaction with streptavidin-horseradish peroxidase conjugate forms a complex that catalyzes dye formation from dye precursor. Wash steps minimize nonspecific background. This format is amenable to multiplexing, permitting internal controls, speciation of bacteria, typing of viruses, and panel testing. An HIV assay performed with this system demonstrated 100% sensitivity and 95% specificity for 64 patients' samples relative to a conventional PCR assay based on 32P solution hybridization. Similarly, an automated closed-vessel assay of cytomegalovirus exhibited 97.5% sensitivity and 100% specificity.
Rare earth cryptates and homogeneous fluoroimmunoassays with human sera
Authors: Mathis G 
Published: January 2020

Abstract
The rare earth cryptates are used as long-lived fluorescent labels. They are formed by inclusion of a europium ion in the intramolecular cavity of a macropolycyclic ligand containing bipyridine groups as light absorbers. The use of fluoride ions in the measuring medium allows a total shielding of the label fluorescence. Here, I have established the conditions under which the signal of these cryptates can be amplified through the use of nonradiative energy transfer, and applied these conditions to homogeneous immunoassays by using allophycocyanin as the acceptor. Because only a low proportion of the cryptate label is involved in the transfer amplification, and because allophycocyanin emission occurs between the europium emission lines, time-resolved measurement of europium and allophycocyanin emission allows real-time correction of the optical properties of the assay medium. These features have allowed the development of a rapid homogeneous immunoassay of prolactin that can detect as little as 0.3 microgram/L (3rd International Standard 84/500).
Diagnostic accuracy of pancreatic enzymes evaluated by use of multivariate data analysis
Authors: Kazmierczak S C , Catrou P G , Van Lente F 
Published: January 2020

Abstract
We analyzed pancreatic enzyme data from 508 patients with suspected pancreatitis by neural network analysis, by an Expert multirule generation protocol, and by receiver-operator characteristic (ROC) curve analysis of a single test result. Neural network analysis showed that use of lipase provided the best means for diagnosing pancreatitis. Diagnostic accuracies achieved by using amylase only, lipase only, and amylase and lipase in combination were 76%, 82%, and 84%, respectively. Use of the Expert rule generation protocol provided a diagnostic accuracy of 92% when rules for single and multiple samplings were combined. ROC curve analysis for initial enzyme activities showed the maximal diagnostic accuracy to be 82% and 85% for amylase and lipase, respectively; use of peak enzyme activities yielded accuracies of 81% and 88%, respectively. The evaluation of laboratory test data should include analysis of the diagnostic accuracy of laboratory tests by multivariate techniques such as neural network analysis or an Expert systems approach. Multivariate analysis should allow for a more realistic assessment of the diagnosis accuracy of laboratory tests because all the available data are included in the evaluation.
Overtraining in neural networks that interpret clinical data
Authors: Astion M L , Wener M H , Thomas R G , Hunder G G , Bloch D A 
Published: January 2020

Abstract
Backpropagation neural networks are a computer-based pattern-recognition method that has been applied to the interpretation of clinical data. Unlike rule-based pattern recognition, backpropagation networks learn by being repetitively trained with examples of the patterns to be differentiated. We describe and analyze the phenomenon of overtraining in backpropagation networks. Overtraining refers to the reduction in generalization ability that can occur as networks are trained. The clinical application we used was the differentiation of giant cell arteritis (GCA) from other forms of vasculitis (OTH) based on results for 807 patients (593 OTH, 214 GCA) and eight clinical predictor variables. The 807 cases were randomly assigned to either a training set with 404 cases or to a cross-validation set with the remaining 403 cases. The cross-validation set was used to monitor generalization during training. Results were obtained for eight networks, each derived from a different random assignment of the 807 cases. Training error monotonically decreased during training. In contrast, the cross-validation error usually reached a minimum early in training while the training error was still decreasing. Training beyond the minimum cross-validation error was associated with an increased cross-validation error. The shape of the cross-validation error curve and the point during training corresponding to the minimum cross-validation error varied with the composition of the data sets and the training conditions. The study indicates that training error is not a reliable indicator of a network's ability to generalize. To find the point during training when a network generalizes best, one must monitor cross-validation error separately.
Molecular nanotechnology
Authors: Fahy G M 
Published: January 2020

Abstract
Molecular nanotechnology involves the ability to manufacture objects to precise atomic specifications. A central postulate is that any structure that can be specified and that does not violate physical law can be built. Three pathways to molecular nanotechnology are proximate probe technology (the use of improvements of the scanning tunneling microscope, STM), biotechnology, and supramolecular chemistry. Combinations of these technologies appear particularly powerful. The biotechnological approach should make it possible to use in vitro translation systems to manufacture polymers containing at least 10 times as many different artificial monomers as there are natural amino acids. These polymers could further adsorb various other molecular devices, and the use of STMs should enable the complexes to be arranged into sophisticated machines, including molecular computers. The implications include pocket superautomated analyzers and the ability to base medical therapy on the biochemical individuality of specific patients.
P34 The effect of sex on the efficacy and safety of dabigatran dual therapy in atrial fibrillation after PCI: a subgroup analysis from the RE-DUAL PCI trial
Authors: Eccleston D , Ten Berg J M , Steg P G , Bhatt D L , Hohnloser S H , De Veer A , Nordaby M , Miede C , Kimura T , Lip G Y H , Oldgren J , Cannon C P 
Published: January 2020

Abstract

Funding Acknowledgements
Boehringer Ingelheim International GmbH

On Behalf
The Re-DUAL PCI Investigators

Background
The RE-DUAL PCI study (NCT02164864) compared dabigatran dual antithrombotic therapy (D-DAT) with warfarin triple antithrombotic therapy (W-TAT) in patients with atrial fibrillation (AF) undergoing percutaneous coronary intervention (PCI). As previously reported D-DAT reduced bleeding compared with W-TAT and was non-inferior with regard to thromboembolic events.
Aim
The aim of this subgroup analysis of RE-DUAL was to assess the relationship between sex and treatment effects of D-DAT and W-TAT on bleeding and thromboembolic outcomes.

Methods
Patients were randomized to receive W-TAT (warfarin, clopidogrel or ticagrelor, and aspirin) or D-DAT (dabigatran 110 or 150 mg twice daily with clopidogrel or ticagrelor; D110- or D150-DAT). Younger patients (aged &lt; 80 yrs. [&lt; 70 yrs. in Japan]) and US patients irrespective of age received D110-DAT, D150-DAT or W-TAT; older patients (aged ≥ 80 yrs. in non-US countries [≥ 70 yrs. in Japan]) received only D110-DAT or W-TAT. Bleeding and thromboembolic outcomes were assessed according to treatment group and by sex (female vs. male).

Results
A total of 2725 patients were randomized; 2070 patients were male (76.0%) and 655 (24.0%) were female. Overall females were older at time of PCI than males (73.2 ± 7.9 vs. 70.0 ± 8.8 years). The mean CHA2DS2-VASc and modified HAS-BLED scores were higher in women at 4.5 and 2.8, respectively, than in men at 3.3 and 2.7, respectively.
For the primary endpoint of major bleeding events or clinically relevant non-major bleeding events, treatment effects of D110-DAT vs. W-TAT were consistent for female and male patients (females: HR 0.69, 95% CI 0.47-1.01, males HR 0.46, 95%CI 0.37-0.59, interaction p-value 0.084). Similarly, consistent treatment effects were seen for the primary endpoint with D150-DAT vs W-TAT in female and male patients (females HR 0.74, 95% CI 0.48-1.16, males HR 0.71, 95% CI 0.56-0.90, interaction p value 0.83).
No interaction between sex and treatment was observed for D110- or D150-DAT vs W-TAT with regard to the composite efficacy endpoint of death, thromboembolic events or unplanned revascularization (interaction p values 0.73 and 0.72, respectively) (figure).

Conclusion
The treatment effect of dabigatran 110 mg and 150 mg dual therapy vs warfarin triple therapy was consistent across sex groups. This suggests that female and male patients with AF undergoing PCI should be treated equally in terms of the dosage of dabigatran selected for dual therapy strategies.
Abstract P34 Figure. Re-DUAL sex subgroup analysis
P93 The usefulness of dual cardiac autonomic nervous modulation assessment for prediction of mortality in patients with relatively preserved left ventricular ejection fraction
Authors: Kinoshita T , Yuzawa H , Wada R , Yao S , Yano K , Akitsu K , Shinohara M , Koike H , Suzuki T , Abe A , Fujino T , Ikeda T 
Published: January 2020

Abstract

Background
Recent guidelines have stated that reduced left ventricular ejection fraction (LVEF) is the gold standard marker for identifying patients at risk for cardiac mortality. Although reduced LVEF identifies patients at an increased risk of cardiac arrest, sudden cardiac deaths (SCDs) occur considerably more often in patients with relatively preserved LVEF. Current guidelines on SCDs risk stratification do not adequately cover this general population pool. Heart rate variability (HRV) and heart rate turbulence (HRT) are non-invasive electrocardiography (ECG)-based techniques capable of providing relevant information on the cardiac autonomic nervous modulation. Although a large body of evidence about autonomic nervous modulation markers has been reported, the usefulness of HRV and HRT parameters for risk stratification in such patients with relatively preserved LVEF has not yet been elucidated.

Purpose
This study aimed to evaluate HRV and HRT parameters for predicting cardiac mortality in patients with structural heart disease (SHD), including ischemic heart disease, dilated cardiomyopathy and valvular heart disease, who have mid-range left ventricular dysfunction (LVD).

Methods
We prospectively enrolled 229 patients (187 men, age 63 ± 13 years) with SHD who have mid-range LVD (LVEF &gt; 40%). HRV and HRT parameters based on 24-hour ambulatory ECG recordings (Fukuda Denshi Co., Ltd., Tokyo, Japan) were evaluated as follows; SDNN, triangular index, high and low frequency HRV, turbulence onset and slope. The primary endpoint was all-cause mortality. Univariate and multivariate Cox regression analysis were used to assess the association between these cardiac autonomic nervous modulation and mortality.

Results
During a mean follow-up of 21 ± 11 months, all-cause mortality was seen in 11 (4.8%) patients. Univariate Cox regression analysis showed that reduced SDNN (&lt;50ms), reduced triangular index (&lt;20ms) and HRT category 2 were significantly associated with the primary endpoint (P &lt; 0.05). When HRT category 2 combined with reduced SDNN, Multivariate Cox regression analysis revealed that this combination more strongly associates with the primary endpoint (hazard ratio =7.91, 95%CI, 1.82-34.2; P = 0.006).

Conclusion
Dual cardiac autonomic nervous modulation assessment which combined HRT and HRV could be a superior technique to predict mortality in patients with relatively preserved LVEF.
P105 Chronic vagal nerve stimulation suppress thirst through decreasing vasopressin secretion in the rats with chronic heart failure
Authors: Zheng C , Li M , Uemura K , Kawada T , Inagaki M , Sugimachi M 
Published: January 2020

Abstract

Introduction
Chronic right cervical vagal nerve electric stimulation (VS) exerted prognostic benefits in heart failure rats. Exploring the mechanism is significant for clinical application. This study examined the effect of VS on arginine vasopressin (AVP) secretion and drinking behavior in the rats with chronic heart failure.

Methods
After a week recovery of myocardial infarction (MI), surviving rats after were randomly instrumented to VS group and sham stimulation (SS) group. A drop sensor was used to record real-time fluid consumption. Blood samples were collected weekly from the 3rd week during the 6 weeks treatment.

Results
Vagal stimulation did not change the daily fluid consumption (Mean ± SE, 76.2 ± 1.1 vs. 77.8 ± 2.1 ml/kg/day, n = 6, P &lt; 0.05). Thirst, which is defined as per drinking volume (PDV), significantly increased during the early weeks after MI (Fig A), while VS significantly suppressed PDV (Fig B is the data of 6th week) by increasing the drinking intervals. The level of plasma AVP was increased to about 2 fold during 6 weeks observation in the SS group, however, VS suppressed this upregulation (Fig C).

Conclusion
These results suggest that VS may activate vagal afferent components which related to inhibition of AVP secretion and then suppressed the augmentation of thirst. Frequent drinking small volume may benefit for homeostasis in CHF.
Abstract P105 Figure. Drinking behavior and AVP