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W G Gray

Publications and source records attributed to W G Gray.

9 recordsLinked to original sources
PubMed2004

Examination of Darcys law for flow in porous media with variable porosity.

Henry Darcy's experimental studies in 1856 of saturated water flowthrough a homogeneous porous medium contained in a vertical column have provided the basis for the quantitative description of fluid flow in a wide variety of both natural and engineered porous medium environmental systems. Extrapolation of Darcy's original observations and conclusions has led to several commonly applied equations used to model flow in porous media. This work examines this original experimental study, summarizes the appropriate mathematical expressions that ensue directly from the data, and indicates expressions in common use that are suggested, but not actually supported, by the data. The paradoxes that exist in the common approaches for the case of a porous medium with a spatiallyvariable porosity are illustrated. A modified form of Darcy's law, and also of the Hubbert potential, is derived based upon fundamental notions of averaging. The modified form of Darcy's law derived here reduces to the conventional form for a homogeneous porous medium.

Gravitation↗
PubMed2003

Thermodynamic fundamental equation of contact lines: selection of independent variables.

In this work we review the development of the generalized theory of capillarity. When considering the theory of contact lines, we find that the theory developed by Boruvka and Neumann (BN) requires significant modifications. Their choice of parameters (independent variables) for a line is insufficient for formulating the fundamental equation. Furthermore, there are differential geometric constraints on these geometric parameters but not included in their analysis. As a result, their independent variables are in fact dependent. To describe the geometry of contact lines properly, we present a new set of parameters from the differential geometry viewpoint and subsequently give the fundamental equation for the thermodynamic system of contact lines.

Journal Article↗
PubMed2001

Bisphenol a binds to the low-affinity estrogen binding site.

Environmental estrogens are suspected of being involved in the current increase in the incidence of human reproductive malfunctions, such as a decrease in male reproductive capacity and an increased incidence of breast cancer in women. The influences of these compounds have been proposed to be mediated through binding to macromolecules, such as estrogen receptor alpha or beta. In this study we examined whether the low-affinity Type II estrogen binding site (Type II EBS), originally identified in the rat uterus, is a possible mediator of environmental estrogens such as bisphenol A (BPA). Analysis of BPA's binding to an enriched fraction of Type II EBS, using a competition assay, indicated that BPA was able to compete with estradiol in binding to this site. At a concentration of 10-15 microM (comparable to that required to induce uterine proliferation), BPA inhibited the binding of estradiol to Type II EBS by greater than 50%. The binding affinity of BPA for the Type II EBS was only 8-10-fold lower than that of the synthetic estrogen diethylstilbestrol. The binding of BPA to Type II EBS appeared specific to BPA, in that endosulfan, another environmental estrogen, failed to displace estradiol from the site. A comparison of the relative binding affinities of BPA for rat uterine estrogen receptor alpha to that of the Type II EBS implies that BPA preferentially binds to the Type II EBS.

Animals↗
PubMed1998

Estrogen receptor alpha requires no accessory factors for high-affinity binding to a consensus response element.

Estrogen receptor (ER) alpha is commonly thought to bind to a consensus estrogen response element (ERE) as a homodimer, but previous experiments have not ruled out the presence of other proteins in the ERalpha/ERE complex. To characterize this interaction in more detail, we overexpressed mouse (m) ERalpha in a baculovirus system, using the selective advantage of the apoptosis inhibitor p35. Recombinant mERalpha possesses the predicted molecular weight and binds 17beta-estradiol and an oligonucleotide containing a consensus vitellogenin ERE with high affinity. Over a wide concentration range of mERalpha protein (0.1-50 nM), only one complex was detected between mERalpha and vitellogenin ERE in gel shift assays. The ratio of E2:vitellogenin ERE bound by mERalpha was close to 2:1, and each complex contained only one ERE. The molecular weight of the complex was determined to be 160 000, very close to that predicted for two mERalpha proteins and one ERE oligonucleotide, therefore providing strong evidence that no other proteins were present. Recombinant mERalpha was purified such that it was the only protein observable by silver stain. Purified mERalpha and mERalpha in a nuclear extract behaved identically in Ferguson analysis, providing more evidence that only mERalpha was binding to the ERE. Purified mERalpha bound vitellogenin ERE with high affinity (Kd = 0. 92 +/- 0.20 nM), indicating that no other proteins are necessary for high-affinity mERalpha interaction with a consensus ERE. To determine whether ERalpha in an estrogen-responsive mammalian tissue behaves the same as the overexpressed mERalpha, we tested rat uterine cytosol by Ferguson analysis. ERalpha in rat uterine cytosol behaved identically to overexpressed mERalpha, suggesting that ERalpha in the uterine extract also binds to DNA predominantly as a homodimer with no additional proteins.

Animals↗
PubMed1998

Phytoestrogens act as estrogen agonists in an estrogen-responsive pituitary cell line.

There is renewed interest in the medicinal value of natural plant products. One group of plant compounds, the phytoestrogens (PE), has been given considerable attention due to their ability to decrease the incidence of certain estrogen-dependent cancers. In this study, we evaluate the effects of PE on estrogen-dependent pituitary tumor cells by using the immortalized pituitary cell line PR1. Several PE were found to be active in PR1 cells, in that they bound to the estrogen receptor (ER), stimulated growth of PR1 cells, and induced an estrogenic response, prolactin secretion. The PE genistein, coumestrol, and zearalenone bound to the ER present in PR1 cells with an affinity 100-times lower than that of estradiol. However, resveratrol, a plant antitumor agent found in grapes, showed no significant binding to the ER. Zearalenone, coumestrol, and genistein were found to induce prolactin secretion and to stimulate growth, whereas resveratrol showed prolactin secretion but no growth stimulation. The estrogenic effects of PE in PR1 cells were ER dependent, in that addition of the antiestrogen ICI-182,780 inhibited prolactin response. Although resveratrol did not bind to the ER or stimulate growth, it induced prolactin secretion in both a dose- and time-dependent manner. The data presented here demonstrate that PE are active in lactotroph cells of the pituitary.

Animals↗
PubMed1996

Identification and characterization of an estrogen-responsive element binding protein repressed by estradiol.

Cytosolic proteins from uteri of 19-day-old rats were analyzed by an electrophoresis mobility shift assay (EMSA) using a 31 base pair DNA probe containing an estrogen-responsive element (ERE) from the vitellogenin A2 gene. EMSA identified three distinct cytosolic protein-DNA complexes that are separable by Q-Sepharose anion exchange chromatography into an estrogen receptor (ER)-containing fraction (150 mM NaCl eluate) and a non-ER-containing fraction (250 mM NaCl eluate). We thus refer to the non-ER fraction as the ERE binding protein (ERE-BP). The ERE-BP-containing fraction was repressed to 40-50% of its normal levels following a single injection of estradiol. In addition, ERE-BP levels were repressed to the same extent (greater than 50%) by day 20 of the rat's gestational period. Examination of the expression pattern of ERE-BP shows that this activity is differentially expressed in both estrogen-responsive and nonresponsive tissues, with the highest levels of expression occurring in the pituitary. We next examined the specificity of ERE-BP binding by competition analysis using DNA sequences corresponding to binding sites of several known transcription factors. ERE-BP was found to be specific for both the ER binding site (ERE) and TATA binding protein binding sites. Furthermore, saturation analysis demonstrated that ERE-BP binds to the ERE and TATA binding protein sequences with an apparent Kd of 1.2 and 0.12 nM, respectively. Partial purification of ERE-BP using three chromatography steps (Q-Sepharose, hydroxyapatite, and Sephacryl S300) followed by sodium dodecyl sulfate analysis indicated the presence of three major protein bands (p102, p81, and p48) as judged by Coomassie staining. UV cross-linking of the ERE-BP/DNA complex followed by sodium dodecyl sulfate analysis-polyacrylamide gel electrophoresis analysis indicates that the 48 kDa band seen in the final, partially purified fraction correlates with the ERE-BP activity. Thus, this study has identified a unique uterine cytosolic protein that binds to the ER binding site and may influence ER binding.

Animals↗
PubMed1994

A low-affinity estrogen-binding site in pregnant rat uteri: analysis and partial purification.

We have identified a low-affinity (type II) estrogen-binding site (EBS) that is expressed at high levels during pregnancy in rat uteri. Although this activity was detectable in nonpregnant rat uteri, it was present in amounts (0.094 pmol/g of uteri) that were severalfold lower than the high-affinity type I estrogen receptor (0.57 pmol/g of uteri). During pregnancy, at 19-20 days of gestation, the low-affinity type II EBS became the major (> or = 88%) estrogen-binding site in rat uteri. The increase in the level of low-affinity EBS (7.9 pmol/g) in uteri was approximately 85-fold with an approximately 20-fold increase in the specific activity (0.39 pmol/mg) of this form, whereas the high-affinity form remained relatively unchanged. We report here a method of purification of type II EBS from pregnant rat uteri and present an analysis of its DNA and steroid-binding properties. Estradiol-binding studies and Scatchard analysis showed that the type II EBS had an apparent estradiol-binding affinity of > or = 24 nM. Gel filtration and SDS/PAGE analysis indicated that the type II EBS was a monomeric 73-kDa protein. The estradiol binding remained apparently uninhibited in the presence of a large excess of tamoxifen, nafoxidine, or dihydrotestosterone. Estradiol, diethylstilbestrol, and quercitin (a type II EBS-specific inhibitor) competed efficiently. The purified low-affinity EBS did not have sequence-specific DNA-binding activity with the estrogen-responsive element, which indicated that it differs in function from the type I estrogen receptor.

Animals↗