PubMed Health⌕ Search

SEARCH · PubMed Health

Results for “Binding, Competitive”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 91 records · Page 5Linked to original sources
PubMed1974

Conventional and dual competitive binding analyses of thyroxine using the Thyopac-4 kit.

A simple method is described for the simultaneous determinations of thyroxine and the effective thyroxine ratio (ETR) using a single kit of Thyopac-4. Thyopac ETR values obtained were 1.00 +/- 0.03 for normal subjects; 1.22 +/- 0.11 for hyperthyroid; 0.87 +/- 0.04 for hypothyroid; and 1.02 +/- 0.05 for pregnant women. Cases with TBG deficiency and the nephrotic syndrome indicated normal values. The value (x) was correlated significantly (gamma = + 0.961) with the free Thyopac index (y), representing a regression equation of y = 71.7x(2) - 88.4x + 24.2. Thyopac ETR was shown to be useful in differentiating normal thyroid states from abnormal, irrespective of changes in T(4)-binding globulin (TBG). Further, the T(4) value obtained simultaneously made it possible to detect changes in the TBG level by noting an inconsistent value for the effective thyroxine ratio.

Binding, Competitive↗
PubMed1986

[Competitive binding assay with dextran-coated charcoal for measurement of sex hormone-binding globulin in breast cancer].

A simplified method for determination of the binding capacity of the sex hormone binding globulin (SHBG-BC) in human serum and its clinical application are described. This method is based on the estimation of the amount of estradiol (E2) bound with SHBG which possessed binding sites with it. After removing the endogenous steroids from the serum-samples by adsorption with dextran coated charcoal, the value of SHBG-BC increased by 18% compared with that of untreated serum. In 48 patients of various ages with breast cancer, the value of SHBG-BC (34.6 +/- 11.1; mean +/- S.D.), expressed as pico mole per 1 ml serum, did not differ significantly from that of 12 healthy control women (29.6 +/- 6.6). However, in postmenopausal women, the value of SHBG-BC in 24 patients with breast cancer (33.4 +/- 12.2) differed from that of 6 controls (26.1 +/- 4.9; p less than 0.05). In addition, estrogen receptors (ER) were measured by using gel-filtration method in cytosols from the tissue of breast cancer. The value of SHBG-BC in 15 ER-positive cases (40.4 +/- 10.1) was significantly higher than that in 13 ER-negative cases (29.3 +/- 8.7; p less than 0.01), but the value of SHBG-BC and ER did not run parallel completely.

Adult↗
PubMed1978

Evaluation of a competitive binding assay for cortisol using horse transcortin.

A non-chromatographic competitive binding assay (CBA) using horse transcortin has been employed in the routine measurement of cortisol in plasma, urine and amniotic fluids. Comparing the values with those of a radioimmunoassay (RIA) or a fluorimetric method (FM) an excellent correlation between the three methods both in plasma and urine has been calculated in normal subjects and in patients with various endocrine disorders. In amniotic fluids, however, there were discrepancies between CBA and RIA. Whereas CBA showed no differences, RIA gave significantly higher values in amniotic fluids of female than of male fetuses. Elevated free plasma cortisol levels observed in patients with prostatic cancer after diethyl stilboestrol diphosphate therapy did not correlate with unconjugated urinary cortisol concentration as measured with CBA and FM. In newborns, a relatively high plasma level found 12 hours after birth was followed by a nadir on the 2nd and 3rd day of life and by an increase until levels of adults on the 5th day of life were reached.

Amniotic Fluid↗
PubMed1993

Identification of a competitive binding component in vitamin D-resistant New World primate cells with a low affinity but high capacity for 1,25-dihydroxyvitamin D3.

Monkeys in a number of different New World primate genera express a form of compensated target organ resistance to steroid hormones, including 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3]. Characterization of these phenotypes has previously relied upon the study of the 1,25-(OH)2D3-receptor (VDR) interaction in cultured dermal fibroblasts from affected primates. In this report, we show that three of these prototypic phenotypes can be faithfully reproduced in previously established cultured cell lines: B95-8, EBV-transformed B lymphoblasts from the marmoset (Callithrix jacchus), a New World primate with recognized vitamin D resistance; OMK, renal tubular epithelial cells from the owl monkey (Aotus trivergatus), a New World primate with an Old World primate-like VDR phenotype; and MLA144, transformed B lymphoblasts from a gibbon (Hylobates), an Old World primate that expresses the wild-type VDR phenotype. The rank order of specific nuclear uptake and binding of [3H]1,25-(OH)2D3 to the VDR was OMK > or = MLA144 >> B95-8. Despite a 7- to 9-fold difference in cellular VDR content according to ligand binding analyses, there was no discernible difference in the internalization constant Kin for specific cellular uptake of [3H]1,25-(OH)2D3 (0.12-0.26 nM) or in the quantity of VDR detected by immunoblot analysis. We now speculate that the discrepancy in VDR quantitation by binding and immunoblot analysis in the B95-8 New World primate cell line results from the presence of an intracellular, vitamin D metabolite binding moiety in this cell line that competes with the VDR for metabolite binding.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Adapting homogeneous enzyme-linked competitive binding assays to microtiter plates.

Recently devised homogeneous enzyme-linked binding assays useful for the rapid detection of carbohydrate structure/content of intact glycoproteins (via use of lectins as binders) and for quantitating given vitamins (e.g., biotin; using soluble binding proteins) are adapted successfully to a microtiter plate reader format. The problem of nonspecific adsorption of the binders and enzyme-saccharide/vitamin conjugates is solved via the addition of Tween 20 to the assay buffer. More convenient and reliable photometric detection of the preferred labeling enzyme, glucose-6-phosphate dehydrogenase (G6PDH), is accomplished by monitoring the rate of generation of reduced thio-NAD (from thio-NAD) at 405 nm instead of NADH (from NAD) at 340 nm. By employing these modifications it is shown that homogeneous enzyme-linked binding assays can be readily adapted to microtiter plates without loss in analytical assay performance. Results further suggest that other homogeneous assays based on G6PDH, including commercial EMIT assays used routinely in clinical chemistry laboratories for detecting drugs of abuse, could, in principle, be run on microtiter plates to significantly enhance sample throughput.

Binding, Competitive↗
PubMed1995

Transcriptional inhibition of the inducible nitric oxide synthase gene by competitive binding of NF-kappa B/Rel proteins.

The activity of the inducible nitric oxide synthase enzyme (iNOS) is tightly controlled, partly at the transcriptional level. We find NF-kappa B/Rel activation (p50-p50 and p50-p65) in RAW 264.7 macrophages after lipopolysaccharide treatment and binding to both NF-kappa B sites in the mouse iNOS promoter. To delineate the importance of NF-kappa B/Rel in iNOS gene transcription, we used an unusually direct approach to try to improve on the antioxidant-treatment or reporter techniques, namely the depletion of NF-kappa B/Rel activity through the use of a phosphorothioate-modified oligonucleotide containing three copies of the NF-kappa B consensus sequence. The reduction in NF-kappa B/Rel activity (particularly that binding to the downstream of the two sites) was associated with a 50% reduction in NO output and a reduction in the quantity of the iNOS protein expressed. These results point to the probability that physiologically relevant NF-kappa B/Rel activators or repressors other than lipopolysaccharide might crucially affect the macrophage NO response.

Amino Acid Oxidoreductases↗
PubMed1985

Competitive binding assay for drugs that interfere with calmodulin function.

A direct 125I-calmodulin-binding system utilizing a particulate cytoskeletal preparation of the neuronal calmodulin-dependent protein kinase type II provides a simple, rapid filtration-based method for assessing anti-calmodulin drug activity. The binding assay avoids potential artifacts due to direct drug effects on the standard phosphodiesterase enzyme assay while obtaining comparable IC50 values for a spectrum of drugs. It also provides a tool to probe mechanisms of action of putative anticalmodulin agents which may help elucidate selective pharmacologic modifications of pathophysiological processes.

Animals↗
PubMed1987

Further probes into quantitative aspects of competitive binding assays: allowance for effects of antigen multivalency in immunoassays.

Effects of antigen multivalency on procedures for the analysis of immunoassays are examined on the basis of a theoretical expression developed in the context of quantitative affinity chromatography [Nichol, L. W., Ward, L. D., and Winzor, D. J. (1981) Biochemistry 20, 4856-4860] but which is also pertinent to antigen-antibody interactions that may be described in terms of a single intrinsic association constant. Quantitative relationships are generated which provide the basis for more rigorous logit-log analyses of radioimmunoassays in which the antigen is multivalent, and an additional, theoretically superior, linear transform of the basic expression is developed. Simulated binding data for a tetravalent antigen system are then used to demonstrate the curvilinearity of the conventional Scatchard plot for such a system despite the homogeneity of binding sites, and the application of the various linear transforms involving logarithmic functions. Of particular interest in that regard is the observation that the traditional logit-log analyses yield linear plots with the predicted slope of unity even though antigen univalence is an implicit assumption in their application. Results obtained in a solid-phase radioimmunoassay of triiodothyronine are then presented to provide, for that system at least, experimental justification of the above-mentioned assumption that the antibody-antigen interactions may be described in terms of a single intrinsic association constant. Finally, an enzyme-linked immunoassay of ferritin is used to illustrate the possibility that a linear Scatchard plot may be obtained with a multivalent antigen under conditions where steric factors restrict participation of an antigen molecule to a single interaction with immobilized antibody.

Antigen-Antibody Reactions↗
PubMed1983

Competitive binding to the cytosolic 2,3,7,8-tetrachlorodibenzo-p-dioxin receptor. Effects of structure on the affinities of substituted halogenated biphenyls--a QSAR analysis.

The proposed mechanism of action of the toxic halogenated aromatics, typified by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), involves the initial binding to a high-affinity, low-capacity, cytosolic receptor protein. Previous studies have shown that several 4'-halo-2,3,4,5-tetrachlorobiphenyls bind to the TCDD receptor and that a lateral substituent on both phenyl rings is required for activity. Using an extended series of eighteen 4'-substituted-2,3,4,5-tetrachlorobiphenyls as probes, the effects of a variable lateral substituent on receptor binding affinity and the induction of aryl hydrocarbon hydroxylase (AHH) in vivo and in rat hepatoma H-4-II E cells have been determined. For most substituents, there was an excellent correlation between the rank-order potency for receptor binding and the rank-order potency for AHH induction. Based on in vitro binding affinities (EC50 values) of the 4'-substituted tetrachlorobiphenyls, a multiparameter regression equation was formulated correlating the binding constants to physicochemical substituent parameters. For thirteen compounds out of the present series, multiple regression analysis of the binding data led to the following equation: log(1/EC50) = 1.53 sigma + 1.47 pi + 1.09HB + 4.08, r = 0.978. The results suggest that halogen substitution on both phenyl rings is not a requirement for binding and that hydrophobic (pi) and electronic (sigma) substituent constants and a variable for hydrogen bond (HB) formation are significant parameters describing relative binding avidities of this series of substituted biphenyls for the TCDD receptor.

Animals↗