bluuebunny/crossref_metadata_2025_split huggingface dataset:
Influence of the hapten-fluorophore bridge on binding parameters in a fluoroimmunoassay for carbamazepine.
Authors: Sidki A M , Landon J , Rowell F 
Published: January 2020

Abstract
Raising antibodies to a hapten (drug or steroid) requires that it be coupled to a carrier protein through a bridge such that the hapten has maximum exposure with minimum changes in its configuration. In raising antisera in six sheep against carbamazepine (CBZ) coupled to bovine albumin, we found that some subpopulations of the antibodies recognized the bridge linking the drug to the carrier protein. To study the influence of the bridge on a fluoroimmunoassay for CBZ, we prepared four tracers by linking the carbamyl nitrogen of CBZ to fluorescein via four alkyl bridges of different lengths and structures. We calculated various binding parameters--including antibody affinity, binding capacity, and heterogeneity index--for each tracer and chose for the final fluoroimmunoassay the tracer that gave the best displacement with CBZ. We then optimized and validated the assay for direct measurement of CBZ in serum or plasma. The antibodies are coupled to magnetizable particles, which greatly facilitates separation and ensures removal of endogenous interferents.
Radioimmunoassay of aspartate aminotransferase isoenzymes in human serum.
Authors: Leung F Y , Niblock A E , Henderson A R 
Published: January 2020

Abstract
We describe the development of a sensitive, specific radioimmunoassay for the cytoplasmic and mitochondrial isoenzymes of human aspartate aminotransferase (L-aspartate:2-oxoglutarate aminotransferase; EC 2.6.1.1). Isoenzymes from human heart tissue were purified to homogeneity and used to raise high-titer antisera in rabbits. We partly purified the antisera by selective column chromatography. The Bolton-Hunter reagent was used to radioiodinate the isoenzymes. The assay requires 100 microL of serum, includes a solid-phase second-antibody separation, and can be completed in less than 3 h. There was no cross reactivity between the two isoenzymes. As little as 5 micrograms (50 pmol) of each aspartate aminotransferase can be measured per liter of serum.
Rapid colorimetry of zinc in seminal fluid.
Authors: Lampugnani L , Maccheroni M 
Published: January 2020

Abstract
This colorimetric method for determination of zinc in seminal liquid is based on the reaction between zinc and 4-(2-pyridylazo)resorcinol in a buffered solution at pH 9.5. It is very simple, fast (approximately equal to 10 min), and economical. It can be used to determine zinc in "whole" semen, seminal plasma, or isolated spermatozoa. We verified its accuracy for sample concentrations up to 0.4 g/L by comparison with a reference method based on atomic absorption spectroscopy of acidic wet-ashed samples. Calcium and magnesium do not interfere, even in concentrations exceeding the physiological values. Between-run precision at a normal concentration of 0.14 g/L is about +/- 7 mg/L; within-run precision at the same concentration is about +/- 2 mg/L.
Use of Alumina, SephadexGlO, and Ion-ExchangeColumnsto Purify Samplesfor Determination of Epinephrine,Norepinephrine,Dopamine, HomovanillicAcid, and 5-Hydroxyindoleacetic Acid in Urine
Authors: Westerink Ben N C , Bosker Fokko J , O'Hanlon James F 
Published: January 2020

Abstract
Vol. 28
p 1746: In line 10 of column one, "700 mL/L" refers to concentrated perchloric acid, which contains 700 g of the acid per liter.
Vol. 30
p 292, column two:
equation 1 should read:
[See equation in the PDF file]
The second part of equation 5 should read "sT = 1.66" (not "166"). Equation 6 should read sRA = [s2BR + s2PWR/Nk]1/2;and the equation at the bottom of the column should read sRA &lt; [s2BR + s2PWR]1/2;. In the sentence just preceding section 3, change "s2WR" to "s2PWR."
p 299: "Corning," three lines from the bottom of column one, should read "Hoechst."
p 428: In Table 1, at the top of the third column: for "3.06" substitute "3.07" and for "3.94" substitute "4.12."
p 501: Column three, line 19: "...suggests that the thyroxin..." should read "...suggests that the immunogen..."
p 521: In this paper (e.g., Figure 3) the ratios reported are by weight. To express them on a molar basis, multiply by 1.10.
p 584: Units for calcium and phosphate should be "µmol/L" not "mmol/L."
p 758: In the equation near the bottom of column one, delete the bar over the x.
p 780: Four lines were omitted from the top of column 1: both comparing well with those of the Amersham analog-based assay for FT4 (7). The reference interval, 10-23 ng/L, clearly differentiated untreated hyper- and hypothyroid from euthyroid patients, only six of 74 results for euthyroid.
Effect of dialysis on interference by cefoxitin with determination of creatinine.
Authors: Kroll M H , Hagengruber C , Elin R J 
Published: January 2020

Abstract
Interference by cefoxitin with determination of creatinine is less with the Technicon SMAC than with other commercial analytical systems. The SMAC assay involves a single-point kinetic method with dialysis, whereas most other commercial methods are multipoint kinetic without dialysis. The apparent creatinine concentration measured for aqueous solutions of cefoxitin was 73 mmol of creatinine per mole of cefoxitin with the SMAC, 135 mmol/mol with a manual method. Furthermore, we determined for the SMAC that the average fraction of creatinine dialyzed was 0.128 and for cefoxitin, 0.064. Thus, the concentration of and interference by cefoxitin in the reaction mixture for SMAC are reduced by half (i.e., the apparent creatinine concentration for cefoxitin with the manual system multiplied by 0.5 is essentially that noted with the SMAC: 68 vs 73). Thus we conclude that the diminished interference be cefoxitin with determination of creatinine by SMAC is primarily ascribable to the dialysis step.
Bilirubin interference in a peroxidase-coupled procedure for creatinine eliminated by bilirubin oxidase.
Authors: Artiss J D , McEnroe R J , Zak B 
Published: January 2020

Abstract
A colorimetric peroxidase-coupled procedure for determination of creatinine in human serum and urine is described. A 30-s sample pre-treatment with bilirubin oxidase eliminates interference from endogenous bilirubin. The 4-aminoantipyrine-2-hydroxy-3,5-dichlorobenzenesulfonate chromogen system of this method is about fourfold more sensitive than current procedures that involve monitoring NAD(P)H and nearly fivefold more sensitive than the traditional picrate procedures. Incorporation of a sample blank eliminates positive interference from endogenous creatine. Results of the proposed procedure are somewhat lower than those of the common kinetic or equilibrium picrate techniques, as would be expected because of the effects of the well-known interferences in the latter methods.
Clinical evaluation of a fully automated chromogenic method for prothrombin time compared with a conventional coagulation method.
Authors: Tripodi A , Mannucci P M 
Published: January 2020

Abstract
The clinical usefulness of a chromogenic method for prothrombin time (PT) determination has been assessed in a wide range of clinical conditions, and it is compared with the conventional clotting method for PT. The new method appears to be as sensitive as the clotting PT to deficiencies of clotting factors of the extrinsic and common pathway, except for fibrinogen. Patients with proven liver disease were correctly diagnosed with a prevalence of abnormal results comparable to that by the clotting PT. Results by the two methods correlated highly (r = 0.96) for normal and congenitally deficient plasmas as well as for plasmas from patients on oral anticoagulant treatment (r = 0.95). High reproducibility (between-assay CV less than 3%) and easy adaptation to centrifugal analyzers make it a suitable candidate to replace the conventional method.
Zymogen activation: a new system for homogeneous ligand-binding assay.
Authors: Blake D A , Skarstedt M T , Shultz J L , Wilson D P 
Published: January 2020

Abstract
In this ligand-binding assay procedure, sensitivity is enhanced by successive generation of enzyme active sites via two zymogens from the blood-coagulation cascade: Factor X and prothrombin. A protease fraction from Russell's viper venom (RVV) acts upon Factor X, initiating a two-step cascade that culminates in generation of thrombin, the activity of which is monitored with a chromogenic substrate. In the model presented here, the analyte of interest, biotin, is covalently coupled to Factor X. In the presence of avidin, a biotin-binding protein, RVV cannot initiate cascade activity; however, the inhibition can be competitively overcome by addition of free biotin to the reaction mixture. In a complete system, the dose-response curve is linear from 20 to 100 nmol of biotin per liter. Such an assay offers improved sensitivity over many radioisotope-independent immunoassay methods, and may be applicable to a wide variety of analytes.
New separation method for monoclonal immunoradiometric assays and its application to assays for thyrotropin and human choriogonadotropin.
Authors: Rattle S J , Purnell D R , Williams P I , Siddle K , Forrest G C 
Published: January 2020

Abstract
We describe a novel separation procedure for immunoradiometric assays involving monoclonal antibodies in which both radiolabeled and capture antibodies are used in solution, the capture antibody being labeled with fluorescein isothiocyanate (FITC). Separation is achieved by incubation with anti-FITC antibodies on magnetic particles. This technique enhances reaction kinetics relative to those of assays in which a solid-phase capture antibody is used, thus allowing faster reaction times and more economic use of the monoclonal antibodies. The use of anti-FITC magnetic solid phase produces an assay having a highly specific separation method, minimal nonspecific binding, and high sensitivity. The method is illustrated by application to assays for thyrotropin and human choriogonadotropin.
Application of the MAST Immunodiagnostic System to the determination of allergen-specific IgE.
Authors: Miller S P , Marinkovich V A , Riege D H , Sell W J , Baker D L , Eldredge N T , Dyminski J W , Hale R G , Peisach J M , Burd J F 
Published: January 2020

Abstract
The MAST Immunodiagnostic Test System was developed to provide a comprehensive, simple means for the in vitro measurement of multiple antigens or antibodies. The first commercial application of the MAST system incorporates several novel features for cost-effective diagnosis of IgE-mediated allergy in a clinical laboratory or a physician's office. The basis of the MAST system is a unique analytical test chamber, which contains cellulose thread as the solid-phase matrix and allows multiple test results from a single assay. This test chamber incorporates both positive and negative controls and requires no volume-dependent pipetting steps. Immunographic exposure onto high-speed Polaroid instant film allows for quantifying results with an automatic recording infrared-transmittance densitometer. Test results are easily interpreted by using a patient test record provided with the system. The MAST system greatly simplifies testing for allergen-specific IgE, while retaining specificity and sensitivity. Currently, with the MAST system one can simultaneously measure picomoles of allergen-specific IgE in up to 35 different allergen classes. In addition to allergy testing, the MAST technology is applicable to other immunodiagnostic profiles.
Liquid chromatography in open tubes.
Authors: Maskarinec M P , Sepaniak M J , Balchunas A T , Vargo J D 
Published: January 2020

Abstract
Liquid chromatography in open tubes is evaluated for applicability to clinical analysis. The principal advantages of this technique include high separation power, reduced solvent consumption, and small sample volume requirements. Technology for the production of columns as small as 8 micron (i.d.) is described. Preliminary results are encouraging and, with further development in column and detector technology, the technique should find wide applicability in clinical analysis.
Novel shell/core particles for automated turbidimetric immunoassays.
Authors: Litchfield W J , Craig A R , Frey W A , Leflar C C , Looney C E , Luddy M A 
Published: January 2020

Abstract
Recent innovations in particle design have led to the development of highly sensitive and reproducible immunoassay methods for the Du Pont aca discrete clinical analyzer. Key advances include the synthesis and use of particles less than 1 micron in diameter with high refractive index cores surrounded by thin, chemically reactive shells. The cores are prepared by emulsion polymerization to a well-defined size that depends on the choice of monomer and the requirements for turbidimetric signal. Methods for measuring therapeutic drugs (e.g., theophylline) involve particles with polystyrene cores; other methods require cores with higher refractive indices such as polyvinylnaphthalene. The shells are critical for overall method performance because they bind covalently the immunochemicals of interest. Polyglycidyl methacrylate shells have been used effectively to attach antigens and haptens to the particle surface.
Affinity-column-mediated immunoenzymometric assays: influence of affinity-column ligand and valency of antibody-enzyme conjugates.
Authors: Freytag J W , Lau H P , Wadsley J J 
Published: January 2020

Abstract
We describe an affinity-column-mediated, enzyme-linked immunometric assay that is highly sensitive and adaptable to automation. Digoxin is the model test analyte. A comparison of digoxin with its analog, ouabain, for use as the immobilized ligand on the affinity column showed ouabain to be superior. We also report the effect of column elution rate. Antibody-enzyme conjugates prepared with the monovalent Fab'-fragment and the divalent F(ab')2-fragment coupled to beta-galactosidase are compared in terms of their overall assay performance. Although the monovalent Fab'--beta-galactosidase conjugate yields a more sensitive assay and dose-response curves that are linear over a wider range, the divalent F(ab')2--beta-galactosidase conjugate provides an assay with adequate sensitivity and extremely good precision, and is generally easier to synthesize reproducibly. This fully automated, rapid, and precise assay for digoxin is compatible with the Du Pont aca discrete clinical analyzer.
Elimination of "hook-effect" in two-site immunoradiometric assays by kinetic rate analysis.
Authors: Hoffman K L , Parsons G H , Allerdt L J , Brooks J M , Miles L E 
Published: January 2020

Abstract
Two-site or sandwich immunoradiometric assays (IRMAs) offer theoretical advantages over competitive immunoassay systems for sensitivity, precision, and rapid incubation. The practical realization of these advantages has been limited by the phenomenon of the "high-dose hook effect," such that high concentrations of an analyte give similar responses to those of much lower concentrations. We have developed a kinetic rate monitoring IRMA system for use with the Kineti-Count 48TM, an automated kinetic radioassay analyzer, which eliminates "hook-effect" interference, thereby permitting optimal assay design for increasing sensitivity and reducing incubation time. Practical illustration of these concepts is demonstrated by a 10-min, automated, quantitative assay we developed for human choriogonadotropin. The assay can detect as little as 1.2 int. units/L and kinetically screens for the hook effect. Kinetic rate analysis of the two-site IRMA and potentially of nonisotopic counterparts permits improvements in the speed and reliability of these immunoassays.
Anti-IgG binding test to assay circulating IgG-containing immune complexes from polyethylene glycol precipitates.
Authors: Levinson S S , Goldman J O , Feldkamp C S 
Published: January 2020

Abstract
We describe a simple approach for assaying immune complexes from serum by using anti-IgG as the indicator after a three-step extraction procedure with polyethylene glycol. Analysis of the data indicates that assays of such extracts for immune complexes by absorbance nephelometry, kinetic light scatter, and immunoradiometric techniques correlate well. For 116 samples, results by absorbance nephelometry correlated (r = 0.86) with those by the C1q-binding test. The present assay and the Raji cell test were more sensitive than the C1q-binding test (p less than 0.001) for detecting increased concentrations of immune complexes in 29 samples from patients with immune-complex-type diseases. The basic approach we describe may lend itself to broad applications for use with various immunoassay techniques.
Clinical effectiveness of the Du Pont aca measurement of creatine kinase MB in serum from patients in a coronary-care unit.
Authors: Leroux M L , Rabson J , Desjardins P R 
Published: January 2020

Abstract
We evaluated the clinical effectiveness of measuring creatine kinase (CK; EC 2.7.3.2) isoenzyme MB and lactate dehydrogenase (LD; EC 1.1.1.27) isoenzymes in diagnosis of acute myocardial infarction. We used an agarose electrophoresis method to measure CK and LD isoenzymes and the Du Pont aca column method to measure CK-MB. Serial blood specimens were drawn from 100 patients consecutively admitted to our Coronary Care Unit. Because of the low diagnostic specificity for CK-MB measurements by both agarose electrophoresis and the discrete-analysis method, as compared with reported values, we re-evaluated our isoenzyme data by using Receiver Operating Characteristic curves. Such analysis of the data established optimal decision levels of greater than or equal to 25 U/L and greater than or equal to 18 U/L plus greater than or equal to 6% of total CK for serum CK-MB measured by the agarose electrophoresis and the aca methods, respectively, and an optimal decision level of greater than or equal to 0.92 for the ratio of LD 1/2 measured after agarose electrophoresis. At these decision levels we obtained a sensitivity of 100%, 100%, and 95% and a specificity of 94%, 92%, and 90% for CK-MB (agarose electrophoresis), CK-MB (aca), and the LD 1/2 ratio, respectively.
International Legal Histories as Orders: An Afterword to Martti Koskenniemi’s Foreword
Authors: Iurlaro Francesca 
Published: January 2020

Abstract
In this article I address the question of what Martti Koskenniemi refers to in his EJIL Foreword as Hugo Grotius’ legal imagination – the type of values he was trying to convey and the strategies he meant to pursue while constructing his idea of an international legal order. As a matter of fact, focusing on such an apparently narrow aspect is not just relevant to those with a historical interest in Grotius. It also tells us something about the inveterate relationship between international law and historiographic practices. What I want to suggest here is that the history of international law is not just an a posteriori critical reflection on the international legal order – a subgenre for lovers of intellectual escapism in search of a distraction from the many problems of the contemporary world – but, rather, that one of the many successful projects of international law was (and still is) the ambition to order the world through histories.
The cutaneous microbiota of bats has in vitro antifungal activity against the white nose pathogen
Authors: Grisnik Matthew , Bowers Olivia , Moore Andrew J , Jones Benjamin F , Campbell Joshua R , Walker Donald M 
Published: January 2020

ABSTRACTSince its introduction into the USA, Pseudogymnoascus destructans (Pd), the fungal pathogen of white-nose syndrome, has killed millions of bats. Recently, bacteria capable of inhibiting the growth of Pd have been identified within bat microbial assemblages, leading to increased interest in elucidating bacterial assemblage-pathogen interactions. Our objectives were to determine if bat cutaneous bacteria have antifungal activity against Pd, and correlate differences in the bat cutaneous microbiota with the presence/absence of Pd. We hypothesized that the cutaneous microbiota of bats is enriched with antifungal bacteria, and that the skin assemblage will correlate with Pd status. To test this, we sampled bat microbiota, adjacent roost surfaces and soil from Pd positive caves to infer possible overlap of antifungal taxa, we tested these bacteria for bioactivity in vitro, and lastly compared bacterial assemblages using both amplicon and shotgun high-throughput DNA sequencing. Results suggest that the presence of Pd has an inconsistent influence on the bat cutaneous microbial assemblage across sites. Operational taxonomic units (OTUs) that corresponded with cultured antifungal bacteria were present within all sample types but were significantly more abundant on bat skin relative to the environment. Additionally, the microbial assemblage of Pd negative bats was found to have more OTUs that corresponded to antifungal taxa than positive bats, suggesting an interaction between the fungal pathogen and cutaneous microbial assemblage.
Implementation of Kenyan comprehensive school health program: improvement and association with students’ academic attainment
Authors: Akiyama Takeshi , Njenga Sammy M , Njomo Doris Wairimu , Takeuchi Rie , Kazama Haruki , Mutua Alex , Walema Barnett , Tomokawa Sachi , Estrada Crystal Amiel , Henzan Hanae , Asakura Takashi , Shimada Masaaki , Ichinose Yoshio , Kamiya Yasuhiko , Kaneko Satoshi , Kobayashi Jun 
Published: January 2020

AbstractThere is growing evidence supporting the effectiveness of a comprehensive school health program. However, implementation in developing countries is a challenge. Furthermore, the available information on the association between a comprehensive school health program and students’ academic attainment is limited. In Kenya, a project to verify the effects of a comprehensive school health program was carried out in Mbita sub-county, Homa Bay County from September 2012 to August 2017. This study aimed to clarify the improvement of school health during the project years and the association between school health and students’ academic attainment. Primary schools in Mbita sub-county were selected as study sites. We assessed 44 schools’ scores on a school health checklist developed from the Kenyan Comprehensive School Health Program, the students’ mean score on the Kenya Certificate of Primary Education (KCPE), and absenteeism during the project years (2013–16). The mean school health checklist score (n = 44 schools) was 135.6 in 2013, 169.9 in 2014, 158.2 in 2015 and 181.3 in 2016. The difference of the mean score between 2013 and 2016 was significant. In addition, correlation analysis showed a significant association between mean KCPE score in the project years and school health checklist score (Pearson’s coefficient was 0.43, p = 0.004). The results of this study suggest improvements of school health by the implementation of the Kenyan Comprehensive School Health Program and students’ academic attainment.
Do constitutional entrenchment clauses matter? Constitutional review of constitutional amendments in Europe
Authors: Hein Michael 
Published: January 2020

Abstract
Do constitutional entrenchment clauses matter? And if yes, how? This article examines these questions by analyzing a comprehensive collection of 154 decisions issued by European constitutional and supreme courts from 1945 up to 2016, on the constitutionality of constitutional amendments. The article shows that entrenchment clauses do matter: in the vast majority of decisions studied, the claimants and/or the courts referred to a constitutional entrenchment clause. About one-fourth of these cases resulted in the invalidation of a constitutional amendment, most of which were based on an “eternity clause,” that is, the most extreme type of entrenchment clauses. However, the article also demonstrates that most of these invalidations can be assessed as instances of democracy-adverse judicial activism. The article concludes, therefore, that entrenchment clauses cannot be considered an unambiguous instrument for the protection of democratic constitutionalism.