bluuebunny/crossref_metadata_2025_split huggingface dataset:
Enzyme immunoassay for c‐reactive protein analysis
Authors: Unten Sharon K. , Hokama Y. 
Published: March 2007

AbstractA solid‐phase enzyme immunoassay method using goat anti‐CRP sensitized to polyvinyl 96 well plates and a second rabbit anti‐CRP highly specific for gamma mobility CRP is described in this study. Examination of serum samples of normal healthy individuals from cord blood, children (2–12 yrs) and adults (16–83 yrs) by this EIA method showed a progressive increase in CRP levels with age. The mean and standard error of the mean in ng/ml CRP was as follows with respect to age distribution: cord blood, 310 ± 54; children (2–12 yrs), 664 ± 87; adults (16–35 yrs), 1,145 ± 116; adults (36–50 yrs), 3,775 ± 605; adults (51–83 yrs), 7,718 ± 1,482. Individuals in the age range of 22–34 yrs with one or more abnormality in the chemistry profile analysis showed CRP levels with a higher x̄ ± S.E.M. (3,615 ± 925). This is in contrast to the normal individuals of the same age range (1,145 ± 116). This EIA method is sensitive, reproducible, and specific. It should be a useful quantitative method for monitoring low level CRP responders such as newborn and cord blood serum.
Solid‐Phase enzyme immunoassay for terminal deoxynucleotidyl transferase (abbott tdt‐eia) in extracts of whole blood and mononuclear cells isolated from bone marrow and blood
Authors: Fairbanks Tod R. , King William J. , Coleman Mary Sue , Finley Paul R. , Fritsche Herbert A. , Hutter John J. , O'Brien John F. , Manderino George L. 
Published: March 2007

AbstractThe Abbott enzyme immunoassay for detecting terminal transferase (TdT‐EIA) was evaluated at four institutions. Extracts of mononuclear cells isolated from bone marrow and peripheral blood of individuals with leukemic and nonleukemic diseases were assayed. In addition, whole blood specimens, in which TdT was extracted from mononuclear cells directly in blood, were assayed. Intrarun, interrun, and between laboratory coefficients of variation ranged from 6.2%–9.3%, 9.4%–11.1%, and 10.9–13.5%, respectively. Since TdT quantitation in whole blood extracts might be adversely affected by blood components and exogenous substances, studies were carried out to determine whether the TdT‐EIA accurately measured TdT in blood. TdT recovery was virtually 100% in blood extracts provided that an extraction buffer, included in the TdT‐EIA kit, was used. Addition of lipids, human gamma globulin, bilirubin, bacterial DNA polymerase, and chemotherapeutic drugs commonly used to treat acute lymphocytic leukemia had no effect on TdT antigenic activity in whole blood specimens. Linear regression analysis at two institutions comparing TdT quantitation using the TdT‐EIA and DNA polymerase assays yielded correlation coefficients of 0.88 and 0.92. Assay results from 1,019 clinical specimens were used to establish expected values for extracts of whole blood and isolated mononuclear cells from blood and bone marrow. The data demonstrate that the TdT‐EIA is a reproducible assay for quantitating TdT levels in clinical specimens.
Twelve‐Protein immunoassay profile on the cobas fara
Authors: Hudson Gail A. , Poulin Sue E. , Ritchie Robert F. 
Published: March 2007

AbstractThe increasing demand for serum protein quantitation poses several problems for the laboratory, including the lack of availability of high‐volume immunochemistry analyzers, the broad concentration range spanned by clinically important proteins, and the difficulty in assay design caused by the variable nature of antiserum. This paper details a profile for 12 commonly requested serum proteins that span a concentration range of three orders of magnitude. The system, which is based on the COBAS FARA centrifugal analyzer, has a capacity of 700 tests per workday while exhibiting performance characteristics that equal or exceed those of pre‐existing techniques. Several features of the FARA and of the assay design that contribute to the high volume capacity are discussed.
Analysis of immunodeficiency in multiple myeloma: Observations and hypothesis
Authors: Pilarski Linda M. , Mant Michael J. , Ruether Bernard A. 
Published: March 2007

AbstractPatients with multiple myeloma suffer from often profound immunodeficiency that seriously compromises both longevity and quality of life. The cellular basis for the immunodeficiency is unknown and the mechanism(s) giving rise to and possibly maintaining it is also unknown. In this review, evidence defining cellular defects in the nonmalignant B and T peripheral blood lymphocyte pool is summarized and discussed in terms of possible mechanisms that could give rise to the abberrant immune phenotype characteristic of many patients. The abnormalities we have defined include a deficiency of mature B lymphocytes, presence of often large numbers of pre‐B cells in the blood, arrested differentiation of B cells into immunoglobulin (lg)‐secreting cells, and abnormal progenitorlike T cells in blood.The specificity repertoire of the remaining B lymphocyte population is heavily skewed toward recognition of idiotypic and shared epitopes on the variable region of Ig, perhaps reflecting immune reaction with the monoclonal myeloma Ig. This may begin as a response to tumor antigen that gradually progresses to autoimmunity. A second subset of B cells present at abnormally high frequency in myeloma patients is that of antigen‐experienced memory B cells, which probably encountered antigen prior to the events leading to frank myeloma. In this instance, the high frequency reflects a decrease in aggregate numbers of B cells, although it seems likely that expansion of memory clones also occurs. This is exemplified by the set of memory B cells specific for tetanus toxoid, which are present at normal number but highly enriched frequency in myeloma PBL. Both the antitumor B cells (anti‐idiotype, and anti‐Ig) and the memory cells specific for environmental pathogens appear to be arrested in their differentiation to Ig‐secretors as evidenced by the lack of serum antibody in patients.The above observations are interpreted in terms of the hypothesis that immunoregulatory events triggered by the monoclonal myeloma Ig establish and maintain the immunodeficiency. We speculate that autoimmune suppressor T cells specific for conserved determinants of Ig prevent pre‐B cell differentiation to slg + B cells, and B cell terminal differentiation to Ig‐secretors. If correct, this analysis could lead to therapeutic approaches for the reversal of the immunodeficiency and perhaps some degree of control over the malignant growth.
Immunoperoxidase staining for calcitonin and calmodulin in the cystic fibrosis lung: A possible new therapeutic strategy in cystic fibrosis
Authors: Wolf Paul , Kilbourn J. P. 
Published: March 2007

AbstractThe presence of calcitonin and calmodulin has been demonstrated by immunoperoxidase staining in formalin‐fixed, paraffinembedded sections of lung from autopsy tissues of patients who died as a result of cystic fibrosis. Calcitonin has been stained and quantitated in solitary endocrine cells, which are increased in number and staining intensity in the cystic fibrosis lung compared to chronic obstructive pulmonary disease (COPD) and normal lungs. Cal modulin was increased in the bronchiolar epithelium and alveolar pneumocytes in cystic fibrosis lung as compared to COPD and normal lungs. Because of our findings, we are proposing a possible new therapeutic regimen for treatment of the pulmonary infection utilizing diuretics and calcium channel blocking agents.
Antibody‐Enhanced enzyme‐amplified determination of methotrexate
Authors: Nashed N. E. , Rechnitz G. A. 
Published: March 2007

AbstractThe sensitivity of potentiometric methotrexate assays can be enhanced by the presence of antimethotrexate antibody, which acts to potentiate the inhibitory effect of methotrexate on enzyme activity. This property is demonstrated in an enzyme‐modified determination of methotrexate with a PCO2 membrane electrode in a cyclic scheme using the enzymes dihydrofolate reductase and 6‐phosphogluconic dehydrogenase.
Homogeneous dry reagent immunoassay strips for the determination of therapeutic drugs in human serum or plasma
Authors: Thompson Stephan G. , Boguslaski Robert C. 
Published: March 2007

AbstractSpecific dry reagent immunoassay strip tests for therapeutic drugs have been developed with the apoenzyme reactivation immunoassay system (ARIS). This simple and convenient method for measuring such analytes in human serum or plasma is described with assays for theophylline, phenytoin, and phenobarbital.
Particle agglutination assay for sm antibodies using purified sm antigen
Authors: Suzuki Hiromasa , Nakamura Robert M. , Tsay Gregory J. , Tan Eng M. 
Published: March 2007

AbstractSm antibodies are specific serum markers for the diagnosis of systemic lupus erythematosus (SLE) and are identified by immunodiffusion (ID), counterimmunoelectrophoresis (CIE), and passive hemagglutination (PHA) in the clinical laboratories. These current methods have certain disadvantages of sensitivity, specificity, and complexity.In this report, Sm antigen free from other nuclear antigens was purified. Synthetic particles were coated with Sm antigen and used to test for Sm antibodies in patient's sera. The particle agglutination test was specific for only Sm antibodies among different antinuclear antisera from patients with systemic rheumatic diseases. The particle agglutination test had the same sensitivity and specificity as the enzyme‐linked immunoassay (ELISA) test for Sm and RNP antibodies. The correlation coefficient between the ELISA quantitation and the titers obtained by particle agglutination was 0.82. The particle agglutination test can be used for rapid screening and titer determination of anti‐Sm antibodies.
Integrated probe for sweat analysis
Authors: Bezegh Klara , Bezegh Andras , Black Philip G. , Janata Jiri 
Published: March 2007

AbstractSolid‐state integrated probe based on ion‐sensitive, field‐effect transistors has been used for determination of sodium chloride in sweat for diagnosis of cystic fibrosis. This probe can yield useful clinical data faster and with higher reliability than the conventional titration test. A battery‐powered instrument that converts the differential current into milliequivalents of sodium chloride has been constructed. It is possible to perform the diagnosis without inducing sweating with pilocarpine. It is suggested that similar probes with multisensing capability can be used for analysis of other surface body fluids.
Novel enzyme immunoassay of anti‐insulin igG in human serum
Authors: Kohno Takeyuki , Ishikawa Eiji , Sugiyama Satoru , Nakamura Syuji 
Published: March 2007

AbstractA novel enzyme immunoassay of anti‐insulin IgG in human serum is described. A serum sample containing anti‐insulin IgG was treated with dextran‐charcoal at pH 6.0 to remove endogenous insulin and subsequently incubated with dinitrophenyl biotinyl nonspecific rabbit IgG‐insulin conjugate. The reaction mixture was further incubated with a rabbit (antidinitrophenyl bovine serum albumin) IgG‐coated polystyrene ball to trap the complex formed between anti‐insulin IgG and the conjugate. After washing to eliminate nonspecific IgG in the test serum, the polystyrene ball was incubated with dinitrophenyl‐L‐lysine to elute the complex. The eluate was incubated with an avidin‐coated polystyrene ball. Finally, the amount of human anti‐insulin IgG in the complex trapped onto the avidin‐coated polystyrene ball was measured by incubation with rabbit (antihuman IgG (γ‐chain)) Fab'‐peroxidase conjugate. This enzyme immunoassay was 1,000‐fold more sensitive than the conventional enzyme immunoassay, in which an insulin‐bovine serum albumin‐coated polystyrene ball was incubated with a serum sample containing anti‐insulin IgG and subsequently with rabbit (antihuman IgG (γ‐chain)) Fab'‐peroxidase conjugate. The principle of the novel enzyme immunoassay can be used to more sensitively measure antibodies for most kinds of haptens and antigens than the conventional enzyme immunoassay.
Evaluation of estradiol binding in human benign prostatic hyperplasia
Authors: Ganesan S. , Bashirelahi N. , Young J. D. 
Published: March 2007

AbstractWe have systematically evaluated the effect of tris/phosphate buffers, ethylenediamine tetraacetate (EDTA), dithiothreitol (DTT), molybdate, and phenylmethylsulfonyl fluoride (PMSF) on the binding of 17β‐estradiol to human BPH cytosol. Competition binding assays demonstrated that the estrogen receptor was highly specific for 17β‐estradiol. There was statistically no difference in binding observed with tris or phosphate buffers. Two classes of binding (high affinity—type I, low affinity—type II) were observed. DTT (1 mM) showed maximum inhibition of type II binding. There was no significant difference in binding observed in the presence of 1.0 mM EDTA. However, 10 mM EDTA stimulated estradiol binding in both saturation analysis and glycerol gradients. Stimulation of estradiol binding by 10 mM EDTA was observed even in the presence of 1 mM DTT. Molybdate (20 mM), a reagent used to stabilize steroid receptors, significantly inhibited estradiol binding. PMSF inhibited estradiol binding at all concentrations studied and the nature of the inhibition appeared to be uncompetitive. A Bmax (maximum number of binding sites) of 84.4 ± 68.5 fmoles/mg protein with a KD (equilibrium dissociation constant) of 2.5 ± 1.7 mM was observed for type I estrogen receptor binding. Our Bmax values are higher than those reported in other studies even after allowing for possible overestimation. In view of the potential importance of estrogen receptors in human prostate, careful reevaluation of estrogen receptors must be undertaken using proper assay conditions.
ELISA assay for colon cancer
Authors: Lawless B. Desales , Lee Yu‐Chen C. , Crook James E. , Clapp Neal K. 
Published: March 2007

AbstractThe high mortality rate for patients with colon cancer results largely from the inability to diagnose the condition in its early stages. The assay described in this report may be useful in early detection of colon cancer. In the study we use the cotton‐top tamarin, Saguinus oedipus, as an animal model for studying human colon cancer. This New World monkey develops a high incidence of colitis and colon cancer. Antibodies that recognize adenocarcinoma cells and antigens from these cells can be detected in the sera of tamarins with colon cancer. The antibodies are not found in sera of healthy animals. In one procedure, tamarin adenocarcinoma cells are used to coat the wells of a microtiter plate. In an alternate procedure, a glycoprotein adenocarcinoma antigen is used. The antigen is extracted from a cultured tamarin colon cancer line by detergent solubilization followed by affinity chromatography. Test sera are examined by adding biotinylated immunoglobulin serum fractions to the coated wells. The wells are washed and an avidin‐enzyme conjugate is added. Positive sera are identified by the addition of the substrate. The presence of the autoantibodies in the sera may be taken as evidence that the animal has colon cancer.
Current status of fluorescent immunoassays
Authors: Robbins Bruce A. , Nakamura Robert M. 
Published: March 2007

AbstractIn 1941 fluorescent compounds were first introduced as immunochemical labels in assay systems. Several forms of quantitative fluorescent immunoassay, which represent a major alternative to radioimmunoassay and enzyme immunoassay, have been developed and are now widely used in many clinical laboratories. This paper discusses these forms and procedures and their advantages over radioimmunoassay and enzyme immunoassay.
Biochemical nature and biological activity of the vascular permeability factor of bovine platelets
Authors: Ashihara Yoshihiro 
Published: March 2007

AbstractBovine vascular permeability factor (BVPF) was purified from the α‐granule acid extract of platelets (BPAE) by ion‐exchange and gel‐filtration chromatography. BVPF is a cationic protein with an isoelectric point of 8.8. As determined by gel filtration using a Superose 12 column, there appear to be three kinds of BVPF with molecular weights of 30,000, 300,000, and > 450,000. After heat treatment of BPAE, however, BVPF was eluted in a fraction with a molecular weight of 30,000. Vascular permeability increased as much as 2.3‐fold with heat treatment of BPAE at 56°C for 30 minutes and was retained even after heat treatment at 80°C. BVPF was also stable to acid or alkaline treatment, but it was sensitive to proteases or thiol reagents, especially pepsin and 2‐mercapto‐ethanol. After injection of BVPF into the dorsal skin of rabbits, enhancement of vascular Permeability was reconfirmed by 3 minutes of observation electron microscopically. One hour later, exudation had increased at the injection sites, and there was associated inflammatory cell infiltration with occasional strands of fibrin.
Potentiometric determination of serum 5′‐nucleotidase
Authors: Ho May Y. K. , Rechnitz Garry A. , Chen Ted K. 
Published: March 2007

AbstractAn ammonia gas sensor was used to determine 5′‐nucleotidase in serum through the ammonia produced from the coupled reactions of 5′‐nucleotidase and adenosine deaminase. Nonspecific hydrolysis of the substrate 5′‐adenosine monophosphate by serum alkaline phosphatase was corrected for by using the inhibitor concanavalin A. The present method is fast and simple, and offers considerable improvements over existing methods. Results obtained using the present method compare well with the spectrophotometric method and are not affected by sample turbidity.
Correlation of serum levels of CA‐549 to disease status in posttreatment serial samples from breast cancer patients
Authors: Bray Kurtis R. , Gaur Pramod K. 
Published: March 2007

AbstractCA‐549 is a high molecular weight, circulating marker for breast cancer defined by its reactivity to monoclonal antibody BC4E 549. CA‐549 has recently been shown to be elevated in the sera of 50% of advanced breast cancer patients (n = 80), as compared to <2.0% of benign breast disease patients (n = 79) or healthy individuals (n = 257).The purpose of this study was to evaluate CA‐549 as a tool for monitoring the status of breast cancer disease. Serial serum specimens (n = 381) from a total of 126 metastasis breast cancer patients were prospectively collected over 12 months and tested for CA‐549 levels by immunoradiometric assay. The average percent increase in CA‐549 levels in the 18 patients with disease progression was 115 ± 103% (range = −27% to 440%), with 16 of 18 showing an increase of >25%. CA‐549 levels decreased by a mean of −40.5 ± 22.5% (range = 17% to −75%) in 12 patients with breast cancer regression, with 8 of 12 patients showing a decrease in serum CA‐549 of >25%. For 18 patients diagnosed with clinically stable disease, the mean percent change in CA‐549 was only 20.4 ± 5.7%, range = (−27% to 24%), with 14 of 18 having changed by <25%. For patients with no evidence of disease, 75 of 78 were CA‐549 negative throughout the study. Three of 78 patients showed borderline positive CA‐549 elevation for at least one specimen. None of these 78 patients showed highly elevated CA‐549.This study confirms the highly specific nature of CA‐549 as a breast cancer marker, and indicates that changes in CA‐549 levels are associated with the clinical course of the disease. Thus this test may be a useful tool to monitor response to treatment and recurrence of breast cancer.
Enzyme inhibitory homogeneous immunoassay for high molecular weight antigen (I)
Authors: Ashihara Yoshihiro , Nishizono Isao , Tanimoto Tetsuji , Tsuchiya Hiromichi , Yamamoto Katsuhiko , Kido Yasuji , Miyagawa Eiji , Kasahara Yasushi 
Published: March 2007

AbstractA highly sensitive homogeneous enzyme immunoassay has been developed for the determination of antigens with high molecular weight. The method presented is based on steric hindrance of the enzyme against an insoluble substrate by antigen binding to antibody: enzyme, conjugate, and consists of α‐amylase from Bacillus subtilis as label with antibody, human α‐fetoprotein (AFP) or human ferritin as ligands, and insoluble substrate. The assay procedure is as follows: sample is mixed and incubated with the conjugate at 37° C for 20 min. Thereafter, the mixture is incubated at 37° C for 20 min following addition of substrate, and the enzyme reaction is terminated by adding the stopper. Then the mixture is centrifuged at 3,000 rpm for 1 minute, and the absorbance of the supernatant is measured at 620 nm. The measurable range for AFP was 5‐200 ng/ml for ferritin, it was 10‐800 ng/ml. In addition, the method was well correlated with radioimmunoassay; i.e., for AFP r = 0.892 and for ferritin r = 0.951. Furthermore, there was no interference during the assay of endogenous a‐amylase in human serum as a result of the use of the inhibitor specific to mammalian amylase which was purified from bacteria.
C‐reactive protein and platelet activating factor complexes induce production and release of interleukin‐1 by human monocytes
Authors: Hokama Y. , Honda S. A. A. , Hanakahi L. A. , Terada K. Y. 
Published: March 2007

AbstractStimulation of interleukin‐1 (IL‐1) in peripheral blood monocytes exposed to C‐reactive protein (CRP) and C‐reactive protein‐platelet‐activating factor (CRP‐PAF) complexes was demonstrated in vitro in this study. Significant synthesis of intracellular IL‐1, 2‐5‐fold increases in the number of adherent monocytes as determined by indirect immunofluorescence using rabbit anti‐IL‐1 and goat antirabbit IgG‐fluorescein isothiocynate, was shown. Both CRP alone (10 μg/ml) and especially CRP‐PAF mixture (10 μg CRP/1 ug PAF/ml) showed maximum stimulation of IL‐1 of 41 and 95 cells/10 field examined, respectively. CRP alone examined in a dose‐response study showed optimum stimulation of IL‐1 production at the 1 μg/ml concentration with 57% of monocytes showing fluorescence. CRP at varying concentrations with a constant amount of PAF (1 μg/ml) showed optimum IL‐1 staining monocytes (63%) with CRP at 10 μg/ml (10 pg CRP/l μg PAF). Similar significant enhanced extracellular IL‐1 production by monocytes exposed to CRP and CRP/PAF mixtures was shown by the mouse thymocyte assay. Supernatants of monocytes exposed to CRP (10 μg/ml) or CRP‐PAF (10 μg/l μg/ml) demonstrated mouse IL‐1 synthesis of 92 and 110 stimulation indices in the thymocyte assay, respectively. It is suggested that CRP and especially CRP‐PAF complexes activate macrophage through their appropriate receptors to production of monokines which modulate the immune system. Thus, CRP appears to function nonspecifically in a behavior reminiscent of specific immunoglobulins.
CA‐549: A new tumor marker for patients with advanced breast cancer
Authors: Demers Laurence M. , Harvey Harold A. , Glenn Jerry D. , Gaur Pramod K. 
Published: March 2007

AbstractA murine monoclonal antibody, BC4E 549, an IgG1 adapted as tracer antibody in a sandwich immunoradiometric assay, was used to detect a circulating breast cancer‐associated antigen, CA‐549, in patients with advanced breast cancer. The study was designed to examine the specificity and sensitivity of CA‐549 measurements as a specific circulating marker for breast cancer. In a population of 100 normal pre‐and postmenopausal women, serum levels ranged from 1–10 μ/ml. In contrast, 38 of 46 patients (83%) with active, advanced‐stage breast cancer had circulating levels in excess of 10 μ/ml, and levels in several patients exceeded 100 μ/ml. Patients on adjuvant therapy with breast cancer in remission, patients with benign breast disease, and patients with cancers of other sites such as colorectal, ovarian, and lung cancer did not show significant elevations (>10 μ/ml, P > 0.001) of this circulating antigen. Measurement of this breast cancer associated antigen may be of use in the evaluation of treatment for patients with advanced stage breast cancer.