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Therapeutic angiogenesis constitutes an alternative treatment for patients with extensive tissue ischaemia in whom primary vascular reconstruction procedures are not feasible or have previously failed. At present vascular endothelial growth factor (VEGF) has been the most widely used angiogenic factor in experimental and human clinical trials. Early clinical data provide evidence that gene transfer of the VEGF gene can achieve beneficial angiogenesis, with minimal side-effects. Ongoing phase III clinical studies will reveal definitive efficacy. 相似文献
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996.
Detection of altered membrane phospholipid asymmetry in subpopulations of human red blood cells using fluorescently labeled annexin V 总被引:20,自引:12,他引:8
The phospholipids of the human red cell are distributed asymmetrically in the bilayer of the red cell membrane. In certain pathologic states, such as sickle cell anemia, phospholipid asymmetry is altered. Although several methods can be used to measure phospholipid organization, small organizational changes have been very difficult to assess. Moreover, these methods fail to identify subpopulations of cells that have lost their normal phospholipid asymmetry. Using fluorescently labeled annexin V in flow cytometry and fluorescent microscopy, we were able to identify and quantify red cells that had lost their phospholipid asymmetry in populations as small as 1 million cells. Moreover, loss of phospholipid organization in subpopulations as small as 0.1% of the total population could be identified, and individual cells could be studied by fluorescent microscopy. An excellent correlation was found between fluorescence-activated cell sorter (FACS) analysis results using annexin V to detect red cells with phosphatidylserine (PS) on their surface and a PS-requiring prothrombinase assay using similar red cells. Cells that bound fluorescein isothiocyanate (FITC)-labeled annexin V could be isolated from the population using magnetic beads covered with an anti-FITC antibody. Evaluation of blood samples from patients with sickle cell anemia under oxygenated conditions demonstrated the presence of subpopulations of cells that had lost phospholipid asymmetry. While only a few red cells were labeled in normal control samples (0.21% +/- 0.12%, n = 8), significantly increased (P < .001) annexin V labeling was observed in samples from patients with sickle cell anemia (2.18% +/- 1.21%, n = 13). We conclude that loss of phospholipid asymmetry may occur in small subpopulations of red cells and that fluorescently labeled annexin V can be used to quantify and isolate these cells. 相似文献
997.
EF Castillo RM López J Rodríguez-Silverio RA Bobadilla and C Castillo 《Fundamental & clinical pharmacology》1998,12(6):584-589
Summary— The aim of the present study was to assess the role of vascular α1D -adrenoceptors in the sympathetic vasopressor response in vivo. Specifically, we evaluated the effect of a selective α1D -adrenoceptor antagonist, BMY 7378 (8-(2-(4-(2-methoxyphenyl)-1-piperazinyl)ethyl)-8-azaspiro(4,5)decane-7,9-dione 2HCI), on the vasopressor response induced by preganglionic (T7 -T9 ) sympathetic stimulation in the pithed rat. The vasopressor response was dose-dependently sensitive to inhibition by intravenous BMY 7378 (0.1, 0.31, 1 and 3.1 mg/kg), doses of 1 and 3.1 mg/kg being equally effective. Like BMY 7378, 5-methylurapidil (0.1, 0.31, 1 and 3.1 mg/kg) antagonized the vasopressor response to spinal stimulation; doses of 1 and 3.1 mg/kg were also equally effective. In combination experiments, BMY 7378 (1 mg/kg, iv) and the α1A -adrenoceptor antagonist, 5-methylurapidil (1 mg/kg, iv), showed an additive effect. The present results demonstrate that the α1D -adrenoceptor subtype plays an important role in the pressor response to sympathetic nerve stimulation in the pithed rat, and confirm the participation of the α1A -adrenoceptor subtype in the same response. 相似文献
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999.
The full-length murine erythropoietin receptor was expressed in Spodoptera frugiperda (Sf9) cells using a recombinant baculovirus vector. Erythropoietin receptor protein production was maximal 48 hours after infection, as determined by metabolic labeling and immunoblotting; receptor protein varied in molecular mass from 62 to 76 kD. Erythropoietin receptors produced in Sf9 cells could be solubilized using CHAPS in a form capable of binding erythropoietin, and the solubilized receptor bound to immobilized Concanavalin A (Con A) and wheat germ agglutinin, as well as to immobilized recombinant human erythropoietin. Analysis of the distribution of erythropoietin receptors in Sf9 plasma membrane and cytosol fractions using lectin affinity chromatography revealed that membrane-bound receptor had a higher apparent molecular mass and contained the bulk of receptors that bound to wheat germ agglutinin. The receptor was purified by sequential affinity chromatography on Con A-Sepharose and immobilized erythropoietin. Erythropoietin receptors expressed in Sf9 cells were inserted into the plasma membrane in the correct orientation, bound 125I-erythropoietin with a single affinity (kD, 330 pmol/L), and were internalized after ligand binding. However, kD varied inversely with the number of cell surface receptors. Solubilized erythropoietin receptors in whole-cell lysates and isolated plasma membranes exhibited high-affinity binding, with kD values of 92 and 57 pmol/L, respectively. Erythropoietin bound to the surface of infected Sf9 cells could be cross-linked to two proteins with molecular masses of 90 and 65 kD using the homobifunctional cross-linker, disuccinimidyl suberate (DSS). Similar results were obtained with solubilized receptors in whole-cell lysates, and both proteins could be immunoprecipitated by an antiserum to the erythropoietin receptor carboxyl-terminal domain. 相似文献
1000.
Prevention by retinoids of azoxymethane-induced tumors and aberrant crypt foci and their modulation of cell proliferation in the colon of rats 总被引:7,自引:6,他引:1
Zheng Y; Kramer PM; Olson G; Lubet RA; Steele VE; Kelloff GJ; Pereira MA 《Carcinogenesis》1997,18(11):2119-2125
Retinoids are proposed chemopreventive agents that inhibit cell
proliferation and induce differentiation. Their ability to prevent
azoxymethane (AOM)-induced aberrant crypt foci (ACF) and tumors and to
modulate cell proliferation was investigated in the colon of male F344
rats. Thirteen retinoids were evaluated for prevention of ACF and two of
them, 9-cis-retinoic acid (RA) and 4-(hydroxyphenyl)retinamide (4- HPR),
were also evaluated for prevention of colon cancer. The retinoids were
administered continuously in the diet starting 1 week prior to the first of
two weekly 15 mg/kg i.p. injections of AOM and for a total of either 5 or
36 weeks in order to evaluate their effect on colonic ACF and tumors. At a
concentration of 1 mmol/kg diet, 2- (carboxyphenyl)retinamide caused the
greatest reduction (57.7%) in the yield of ACF. 9-cis-RA was toxic at 1
mmol/kg so that it was evaluated at 0.1 mmol/kg, resulting in a 41.6%
reduction in ACF. The ability of the retinoids to reduce the proliferating
cell nuclear antigen (PCNA) labeling index in ACF and in non-involved
crypts correlated with their ability to prevent ACF. Both 9-cis-RA (0.1 and
0.2 mmol/kg diet) and 4- HPR (1 and 2 mmol/kg diet) were highly effective
in decreasing the yield of AOM-induced colon tumors. In summary, retinoids
were demonstrated to reduce cell proliferation and to prevent ACF and
tumors in the colon, suggesting promise as preventive agents for colon
cancer.
相似文献