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Biomedical subjects

E M Wilson

Publications and source records attributed to E M Wilson.

At least 163 records · Page 9Linked to original sources
PubMed1982

Regional and splenic lymphocyte proliferative responses of mice exposed to normal or irradiated Schistosoma mansoni cercariae.

Developing larvae of Schistosoma mansoni migrate through various tissues en route to the liver and mesenteric veins of their definitive host. Regional (lymph node) and systemic (spleen) blastogenic responses to cercarial, adult and egg antigens were measured in CBA/J mice at various times after exposure to normal or irradiated S. mansoni cercariae. Among the separate lymph node groups studied were those draining the tail, thoracic region, intestines, head and neck, and the pelvis. Blastogenic responses were assayed by a micromethod requiring 10(5) cells in 20 microliter volumes per culture. Up to 5 weeks post-cercarial exposure the pattern of responses in lymphoid tissues of infected mice coincided with the migratory route of the parasites. Following oviposition, cellular reactivity was pronounced in all lymph node groups. The reactivity of mice exposed to irradiated cercariae followed a pattern suggestive of a sustained antigenic stimulus only in the nodes draining the tail and lungs. Splenic (systemic) reactivity was roughly comparable between the two exposure groups. These data show the independence and vast differences in the host regional responses following normal or irradiated cercarial exposure.

Animals↗
PubMed1982

Transferrin in the rat prostate Dunning tumor.

A major protein of the rat Dunning prostate tumor has been purified. It has physicochemical properties and an amino acid composition similar to that of transferrin. Furthermore, the isolated tumor protein reacts with antiserum to authentic rat transferrin. Immunoperoxidase staining with rabbit anti-rat transferrin localizes transferrin within tumor acinar glands. Rocket immunoelectrophoresis indicates that transferrin constitutes 30 to 40% of tumor fluid protein, but accounts for only approximately 9% of total serum protein. In normal rat prostate cytosols, the level of transferrin is at least 200 times lower than in tumor cytosol. Nevertheless, dorsal and lateral prostate show variable peroxidase staining indicating the presence of immunoreactive transferrin within acinar glands of these normal tissues. While intense staining for transferrin was found in the interstium of all regions of the normal prostate, transferrin was not detected within acinar glands of coagulating gland, ventral prostate, or seminal vesicle. Immunocytochemical localization of albumin indicates a distribution similar to that of transferrin in normal and neoplastic rat prostate. However, unlike transferrin, the albumin content was lower in tumor fluid than in serum. It is suggested that the high level of transferrin in tumor fluid may be due to selective uptake by the tumor from serum.

Albumins↗
PubMed1981

Strain differences in lymphocyte responses and in vitro suppressor cell induction between Schistosoma mansoni-infected C57BL/6 and CBA mice.

Splenic lymphocyte blastogenic responses to mitogens and antigens were compared in Schistosoma mansoni-infected C57BL/6 and CBA mice. At 1 and 2 weeks of infection, elevated responses to phytohemagglutinin (PHA) were noted in C57BL/6, but not in CBA cultures. During the same period, elevated responses to lipopolysaccharide were observed in cultures from CBA, but not from C57BL/6 mice. Responses to both mitogens were depressed in both strains later in infection. The magnitude of antigen-specific responses was greater in CBA than in C57BL/6 cultures. A cercarial extract (CE) induced significant suppressor cell activity in normal C57BL/6 cell cultures as assessed by inhibition of syngeneic cell responsiveness to PHA. CE did not induce suppressive activity in CBA cultures. CE-induced suppression was unaffected by adherent cell removal, but was sensitive to anti-Thy 1.2 plus complement treatment. CE also directly inhibited PHA-stimulated C57BL/6 cell cultures, but enhanced cultures of CBA mice. The significance of these findings is discussed in relation to the relative strain differences seen in antigen-specific lymphocyte proliferation and the levels of protection induced after immunization with irradiated cercariae.

Animals↗
PubMed1981

Identification of an 8S androgen receptor-promoting factor that converts the 4.5S form of the androgen receptor to 8S.

A protein has been identified that reconstitutes the 4.5S androgen receptor to the classical 8S form on sucrose gradients of low ionic strength (25 mM KCl and 50 mM Tris). Rat prostate Dunning tumor (R3327) cytosol labeled with [3H]dihydrotestosterone was chromatographed on phosphocellulose to separate the 4.5S receptor from this protein, which we refer to as 8S androgen receptor-promoting factor. The 8S promoting factor has the following physiocochemical properties: heat labile (60 C; 30 min), Stokes radius of 58 degrees A, molecular weight of 170,000 or more, precipitates in 40% saturated (NH4)2SO4, elutes from DEAE-Sepharose in 0.1 M KCl, and elutes from phosphocellulose in 0.1 M KCl. The reconstituted 8S receptor complex is similar to the native 8S receptor in that it is labile to heat and physiological salt concentrations, has a Stokes radius of 91 degrees A, and has a molecular weight of approximately 326,000. The 8S promoting factor is present in mature male rat serum, but is undetectable in sera of male rats 16 days of age or younger. The factor appears to be produced by androgen-responsive cells, since it was found in all tissues of the 15-day-old male rat known to contain androgen receptor. Spleen was found to lack both the 8S promoting factor and the androgen receptor. The 8S promoting factor was detected in serum of female rats and in hypophysectomized (44 days) or castrated (2 or 4 weeks) mature male rats. Salt extracts of purified nuclei from the androgen-dependent Dunning tumor also contain the factor. It is suggested that a specific interaction between the two intracellular proteins, 8S androgen receptor-promoting factor and the androgen receptor, may modulate the androgen responsiveness of target cells.

Adenocarcinoma↗
PubMed1980

Biochemical homology between rat dorsal prostate and coagulating gland. Purification of a major androgen-induced protein.

The anatomically distinct organs, rat dorsal prostate and coagulating gland, were found to display remarkable homology in protein composition, including two major androgen-dependent secretory proteins, referred to as dorsal proteins I and II. Dorsal protein I has been purified and found to be a dimer composed of two identical subunits with sedimentation coefficient 4.6 S, Stokes radium 32 A, and Mr = 71,000 (62,000 by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis). This dimer (6.8 S, 46 A, Mr = 150,000) dissociates at high ionic strength and can be partially reconstituted by removal of salt. Dorsal protein I is a basic protein (pI 9) with high lysine content and binds to phosphocellulose but not to DEAE-Sepharose. Schiff's staining shows that it contains carbohydrate. Quantitative rocket immunoelectrophoresis using a rabbit antiserum indicates dorsal protein I is produced only in dorsal prostate and coagulating gland. The protein constitutes approximately 25% of total cytosol protein in both organs, yet makes up on 5% of coagulating gland luminal fluid and ejaculated seminal fluid. It was not detected in the rat dorsal prostate tumor (Dunning R3327H). Dorsal protein II is a larger protein (Mr = 80,000 by SDS gel electrophoresis) with higher carbohydrate content. Under nondenaturing conditions, it has a Stokes radius of > 200 A, corresponding to Mr of > 300,000. Dorsal protein II represents a smaller proportion of total cytosol protein than does dorsal protein I, yet is the predominant protein of coagulating gland fluid. The proteins will be useful probes for studies on androgen regulation of specific gene expression and prostatic secretory mechanisms.

Amino Acids↗
PubMed1979

Effects of proteases and protease inhibitors on the 4.5 S and 8 S androgen receptor.

The size of androgen receptors from rat ventral and dorsal prostate, dorsal prostate (Dunning) tumor, testis, epididymis, and seminal vesicle was determined using Sephadex G-200 chromatogrpahy and sucrose gradient centrifugation. The protease inhibitor diisopropyl fluorophosphate (DFP) was used to minimize receptor breakdown. An 8-9 S, 85 to 106 A receptor (Mr = 280,000 to 365,000; f/fo = 1.9 to 2.4) observed in unfractionated cytosol prepared in low ionic strength buffer with or without DFP is in equilibrium with a 4.5-5 S, 58 A form (Mr = 117,000; f/fo = 1.8) observed at salt concentrations greater than 0.1 M KCl. Receptor partially purified using (NH4)2SO4 or phosphocellulose chromatography in the absence of DFP was present as smaller fragments of 3.6 S, 37 A and 3.0 S, 23 A. Similar fragments could be generated from the 4.5 S or 8 S receptor by mild trypsin treatment. In addition, ventral prostate contains a DFP-insensitive enzyme which specifically converts the 4.5 S, 58 A receptor to the 3.6 S 37 A fragment. The DFP-insensitive enzyme is partially inhibited by rabbit bile and appears similar to the enzyme seminin, a secretory protein of human prostate. Androgen receptor isolated in the presence of DFP from nuclei labeled in vivo is predominantly 4.5 S, 58 A, with smaller forms (37 and 23 A) appearing in the absence of DFP. The 4.5 S, 58 A nuclear receptors were also in equilibrium with a large 8 S form. Receptor breakdown by DFP-insensitive and sensitive proteases appears to be an in vitro phenomenon. Furthermore, the size of the androgen receptor is not significantly changed during receptor migration from cytoplasm to nucleus.

Animals↗
PubMed1979

Characterization of different forms of the androgen receptor.

Three forms of the androgen receptor have been characterized at high ionic strength in partially purified cytosol and nuclear fractions of rat ventral prostate, epididymis, testis, and seminal vesicle by ion exchange chromatography on phosphocellulose, gel filtration in Sephadex G-200, and sedimentation in sucrose gradients. The three forms have the following properties, respectively: elution from phosphocellulose at 0.15--0.20, 0.3--0.5, and 0.20--0.32 M KCl; gel filtration radii of 53, 36, and 22 A; sedimentation coefficients of 5.0S, 3.6S, and 3.0S; frictional ratios of 1.65, 1.4, and 1.0; and molecular weights of 115,000, 55,000, and 29,000. In testis and epididymis cytosol, the 53 A, 5S form was more abundant than the 36 A, 3.6S form. In ventral prostate, the 36 A, 3.6S receptor was the predominant form. Variable amounts of all three forms were observed in seminal vesicles. Conversion from 36 A, 3.6S to 22 A, 3.0S was induced by heating ventral prostate cytosol and could be blocked by serine protease inhibitors. The 22 A, 3.0S receptor fragment from heated prostate cytosol was similar in size and symmetry to receptor extracted in 0.5 M KCl from prostate nuclei labeled in vivo. Extracts of epididymis and seminal vesicle nuclei contained both 36 and 22 A forms.

Animals↗
PubMed1979

Regional cerebral blood flow comparison of right and left hand movement.

Regional cerebral blood flow measurements were made in normal right-handed subjects by the 133 xenon inhalation method at rest and during movement of either hand. Left hand movement evoked a prominent focal flow increase in the right hemisphere in the Rolandic region. During right hand movement, there was a smaller and not statistically significant increase in left hemisphere Rolandic region. This suggests that more effort is required of right-handed subjects to move the left hand than the right, or that motor organization differs for left and right hand movements. Of several flow indices tested, there was little difference among gray matter flow rate, the product of gray matter flow and relative weight, the mean regional flow, and the initial slope index, all reflecting the flow changes with nearly equal sensitivity. The gray matter weight and fractional flow did not change consistently during hand movement.

Adult↗
PubMed1977

9S binding protein for androgens and progesterone.

A steroid binding protein fraction with a sedimentation coefficient of approximately 9 S (molecular weight approximately equal to 200,000) has been identified in 105,000 X g supernatants of several androgen-responsive organs. Highest concentrations were found in epididymis and testis, but small amounts were detected in prostate, seminal vesicle, kidney, submandibular gland, and lung. The 9S protein binds [3H]dihydrotestosterone (17beta-hydroxy-5alpha-androstan-3-one) and [3H]progesterone (4-pregnene-3,20-dione) with equilibrium binding constants of approximately 10(5) M-1 and 10(6) M-1, respectively. The concentration of 9S binding sites in epididymis is approximately 10(-11) mol/mg of supernatant protein, which is at least 10(5) times greater than the concentration of androgen receptor. 9S binding protein appears to be a nonsecretory, intracellular protein and has properties different from the andorgen receptor. It is unretarded on DEAE-Sephadex chromatography at pH 8.0, and its sedimentation rate on sucrose gradients is not altered at high ionic strength (0.4 M KCl). Like the androgen receptor, its binding activity, which is maximal between pH 7 and 9.5, is heat labile, decreased by sulfhydryl reagents, and enhanced by 2-mercaptoethanol. It is suggested that because of its high concentration and low affinity, 9S binding protein may function in the intracellular accumulation of compartmentalization of androgens or progesterone.

Androstane-3,17-diol↗
PubMed1977

Stimulation by human placental conditioned medium of hemopoietic colony formation by human marrow cells.

Medium conditioned by human placental tissue was found to stimulate granulocytic and monocytic colony formation by human marrow cells in semisolid agar cultures. The colony-stimulating activity of unfractionated conditioned medium was equivalent to the activity of standard peripheral blood underlayers. Placentas were a reliable source of active material, and one placenta provided enough material to stimulate 5,000-10,000 cultures of normal or leukemic cells. The colony-stimulating factor in human placental conditioned medium (CSFHPCM) was concentrated and purified 1800-fold using ammonium sulfate precipitation, calcium phosphate gel absorption, DEAE-cellulose batch absorption, gel filtration on Sephadex G-150, and polyarcylamide gel gel electrophoresis. The active factor behaved on gel filtration as a macromolecule with an apparent molecular weight of 30,000 daltons. The active factor in placental conditioned medium was not dependent on the presence of adherent marrow cells with endogenous colony-stimulating activity.

Bone Marrow↗
PubMed1976

Binding properties of androgen receptors. Evidence for identical receptors in rat testis, epididymis, and prostate.

Androgen receptors in crude and partially purified 105,000 X g supernatant fractions from rat testis, epididymis, and prostate were studied in vitro using a charcoal adsorption assay and sucrose gradient centrifugation. Androgen metabolism was eliminated during receptor purification allowing determination of the kinetics of [3H]-androgen-receptor complex formation. In all three tissues, receptors were found to have essentially identical capabilities to bind androgen, with the affinity for [3H] dihydrotestosterone being somewhat higher than for [3H] testosterone. Equilibrium dissociation constants for [3H] dihydrotestosterone and [3H] testosterone (KD = 2 to 5 X 10(-10) M) were estimated from independently determined rates of association (ka congruent to 6 X 10(7) M-1 h-1 for [3H] dihydrotestosterone and 2 X 10(8) M-1 h-1 for [3H] testosterone) and dissociation (t 1/2 congruent to 40 hr for [3H] dihydrotestosterone and 15 h [3H] testosterone). Evaluation of the effect of temperature on androgen receptor binding of [3H]testosterone allowed estimation of several thermodynamic parameters, including activation energies of association and dissociation (delta H congruent to 14 kcal/mol), the apparent free energy (delta G congruent to -12 kcal/mol), enthalpy (delta H congruent to -2.5 kcal/mol), and entropy (delta S congruent to 35 cal col-1 K-1). Optimum receptor binding occurred at a pH of 8. Receptor stability was greatly enhanced when bound with androgen. Receptor specificity for testosterone and dihydrotestosterone was demonstrated by competitive binding assays. The potent synthetic androgen, 7 alpha, 17 alpha-dimethyl-19-nortestosterone, inhibited binding of [3H] testosterone or [3H] dihydrotesterone nearly as well as testosterone and dihydrotestosterone while larger amounts of 5 alpha-androstane-3alpha, 17 beta-diol and nonandrogenic steroids were required. Sedimentation coefficients of androgen receptors in all unfractionated supernatants were 4 and 5 to 8 S. Differences in sedimentation coefficients were observed following (NH4)2SO4 precipitation which did not influence the binding properties of the receptors. These results, together with measurements of3alpha/beta-hydroxysteroid oxidoreductase activity in vitro, suggest that organ differences in receptor binding of [3H] dihydrotestosterone and [3H] testosterone in vivo result from relative differences in intracellular concentrations of these androgens rather than from differences in receptor affinities.

Androgens↗