Biomedical subjects

M R Hernandez

Publications and source records attributed to M R Hernandez.

60 records · Page 4Linked to original sources

PubMed1982

Specific estrogen receptors in human gingiva.

The cytoplasm of normal human male and female gingiva contains a receptor capable of specifically binding 17 beta-estradiol and moxestrol (R-2858) with high affinity (Kd = approximately 3.4 X 10(-10) M) and low capacity (4.5 fmol/mg protein). The binding is sensitive to heat (destroyed by warming to 37 C for 60 min), proteolytic enzymes (pronase, trypsin, and chymotrypsin), and exhibits a pattern of competition similar to that obtained with estrogen receptors from other target tissues. Nuclear uptake of [3H]estradiol was demonstrated by using a dry autoradiographic technique. Specific nuclear localization of [3H]estradiol was found predominantly in basal and spinous layers of gingival epithelium, stromal connective tissue cells (fibroblasts), and endothelial cells and pericytes of small blood vessels in the lamina propria. There was no difference between the Kd values in normal and diseased tissue or between the Kd values or number of binding sites and the age or sex of the patient. However, there was a difference between the amount of estrogen binding sites per mg protein in normal tissue compared to gingiva with dilantin hyperplasia. These results provide the first direct evidence that human gingiva may function as a target organ for estrogens.

Adolescent↗

PubMed1982

Glucocorticoid receptor binding in bovine lens.

The present study demonstrates glucocorticoid receptor localization in the nuclei of cells of the anterior epithelium and bow region and its absence in all other regions of the lens. Both autoradiographic and biochemical studies show that triamcinolone acetonide is inactive whereas tetrahydrocortisol (previously considered to be an inactive metabolite of cortisol) is an active competitor of dexamethasone binding to the glucocorticoid receptor in the bovine lens. The consistency of these findings by both techniques supports the validity of this observation. In addition, it indicates that autoradiography can be used for studying glucocorticoid agonist/antagonist relationships in the human lens, where only small amounts of tissue are available.

Animals↗

PubMed1981

Corneal-conjunctival uptake of topical 3H-dexamethasone in the rabbit eye.

Topically administered 3H-dexamethasone was localized radioautographically to the nuclei of several cell types in the cornea and conjunctiva. In the cornea, label was found in the epithelium and keratocytes. In the conjunctiva, label was found in the epithelium and in the connective tissue cells and blood vessel endothelial cells of the stroma. Prior administration of a 350-fold excess of nonlabeled hormone completely suppressed this localization, indicating that these cells are specific glucocorticoid target cells. There was no localization found in the corneal endothelium. Accumulation of silver grains between corneal stroma and Descemet's membrane suggests that this membrane acts as a partial barrier to diffusion of the steroid.

Animals↗

PubMed1980

Glucocorticoid localization by radioautography in the rabbit eye following systemic administration of 3H-dexamethasone.

Dexamethasone was localized radioautographically in the nuclei of target cells of the rabbit eye following intravenous administration of the labeled steroid. Specifically bound steroid was found in the nuclei of stromal and endothelial cells of the outflow pathway region. This suggests that the glucocorticoid-induced alteration in outflow facility may be mediated by specific effects in these target cells. Nuclear localization was also found in conjunctiva, iridial smooth muscle, choroidal stroma, retina, and sclera, suggesting a physiologic role for glucocorticoids in these tissues as well.

Animals↗

PubMedDate not supplied

Effect of anticoagulants on activation of polymorphonuclear leukocytes induced by shear stress.

We analyzed how actin polymerization, CD11b expression and homotypic aggregation could be used as markers to study leukocyte activation. Leukocytes were obtained from blood anticoagulated with: citrate, unfractioned heparin (UH) and low molecular weight heparin (LMWH). Flow cytometry was used to study actin polymerization and expression of CD11b after leukocyte exposure to shear stress. Leukocyte aggregation was microscopically assessed. Shear increased both actin polymerization and expression of CD11b in citrated blood (100.1±7.1 vs. 85.8±8.5 p< 0.05 and 53.5±3.5 vs. 20.7±5.1; p< 0.005 respectively). These parameters remained unmodified in UH samples. Using both anticoagulants together, we observed increase in CD11b expression induced by shear stress (59.3±2.1 vs. 25.1±11.0; p< 0,05). LMWH samples showed higher basal levels of actin polymerization and CD11b expression than citrated samples (237±40.8, vs. 85.8±8.5 p< 0.05 and 47.8±2.6, vs. 20.7±5.1; p< 0.005) but no changes induced by shear were observed. When LMWH was used in combination with citrate we observed a decrease in basal activation and significant modifications in CD11b expression induced by shear stress (80.0±4.1 vs. 50.4±2.7). Leukocyte aggregation was modified by UH at basal levels and by LMWH after shear stress. These results indicate that exposure to shear stress results in leukocyte activation. The choice of anticoagulant is a crucial factor in studies of leukocyte function.

Journal Article↗

PubMedDate not supplied

Inhibition of fibrin deposition on the subendothelium by a monoclonal antibody to polymorphonuclear leukocyte integrin CD11b. Studies in a flow system.

BACKGROUND AND OBJECTIVE: The development of prothrombotic and procoagulant states may be regulated by direct platelet-leukocyte contact mediated by membrane receptors. We have investigated the role of CD11b integrin in polymorphonuclear leukocytes (PMN) on fibrin formation and platelet reactivity with vascular subendothelium. METHODS: Studies were carried out following a well-established perfusion model, employing either citrated blood, where fibrin formation is blocked, or blood anticoagulated with low molecular weight heparin, which allows thrombin and fibrin formation. Isolated PMN or platelets were treated with specific monoclonal antibodies to CD11b or to CD62P, respectively, and incorporated in reconstituted blood. RESULTS: Treatment of PMN with anti-CD11b significantly decreased the percentage of surface covered with a thick layer (> 5 microns) of fibrin (34.8 +/- 3.3% vs 53.3 +/- 4.9% in control, p < 0.05); it also reduced the average height of fibrin layer and the number of adherent leukocytes (7.9 +/- 1.2 microns vs 10.6 +/- 1.4 microns in control, p < 0.05; and 87 +/- 8 PMN/mm2 vs 186 +/- 25 PMN/mm2 in control, p < 0.05) respectively. Treatment of PMN with CD11b did not significantly affect the attachment of platelets onto the subendothelium when using citrated blood, though a slight decrease in platelet adhesion was observed in the heparinized samples. Treatment of platelets with anti-CD62P did not significantly modify any of the parameters studied. INTERPRETATION AND CONCLUSIONS: Our results indicate that PMN have a role in promoting fibrin deposition under flow conditions, through the participation of CD11b integrin. Under our experimental conditions, this effect does not seem to be influenced by CD62P expressed on activated platelets.

Animals↗