RESUMO
PURPOSE: Nanotechnology is an emerging field with significant translational potential in medicine. In this study, we applied gold nanoparticles (GNP) to enhance radiation sensitivity and growth inhibition in radiation-resistant human prostate cancer cells. METHODS: Gold nanoparticles (GNPs) were synthesized using HAuCl4 as the gold particle source and NaBH4 as the reductant. Either thio-glucose or sodium citrate was then added to the solution separately to bind the GNPs to form thio-glucose-capped gold nanoparticles (Glu-GNP) and neutral gold nanoparticles (TGS-GNPs). Human prostate carcinoma DU-145 cells were exposed to vehicle, irradiation, 15nM TGS-GNPs, or 15nM Glu-GNPs, or GNPs plus irradiation. The uptake assays of GNP were performed using hemocytometer to count cells and the mass spectrometry was applied to calculate gold mass. The cytotoxicity induced by GNPs, irradiation, or GNPs plus irradiation was measured using a standard colorimetric MTT assay. RESULTS: Exposure to Glu-GNPs resulted in a three times increase of nanoparticle uptake compared to that of TGS-GNPs in each target cell (p < 0.005). Cytoplasmic intracellular uptake of both TGS-GNPs and Glu-GNPs resulted in a growth inhibition by 30.57% and 45.97% respectively, comparing to 15.88% induced by irradiation alone, in prostate cancer cells after exposure to the irradiation. Glu-GNPs showed a greater enhancement, 1.5 to 2 fold increases within 72 hours, on irradiation cytotoxicity compared to TGS-GNPs. Tumour killing, however, did not appear to correlate linearly with nanoparticle uptake concentrations. CONCLUSION: These results showed that functional glucose-bound gold nanoparticles enhanced radiation sensitivity and toxicity in prostate cancer cells. In vivo studies will be followed to verify our research findings.
Assuntos
Ouro/farmacologia , Nanopartículas Metálicas/efeitos da radiação , Neoplasias da Próstata/radioterapia , Tolerância a Radiação , Sobrevivência Celular/efeitos da radiação , Ouro/farmacocinética , Humanos , Masculino , Neoplasias da Próstata/patologiaRESUMO
OBJECTIVES: We evaluated the effect of the St Jude Medical sutureless anastomotic connector on endothelium-dependent and -independent saphenous vein graft relaxation, as well as on clinical outcomes and graft patency in patients. METHODS: Human saphenous vein grafts were assigned to control or connector groups (loaded for 1 or 5 minutes; n = 18). Isometric dose-response curves to endothelium-dependent and -independent (sodium nitroprusside) vasodilators were constructed in saphenous vein grafts precontracted with phenylephrine. Thrombin-mediated vasorelaxation, an early determinant of saphenous vein graft failure, was also evaluated. Percent maximum relaxation was compared between groups. Patients in whom the St Jude Medical connector was employed underwent clinical follow-up, stress tests, and angiography 6 to 12 months postoperatively. RESULTS: A23187-induced endothelium-mediated relaxation, sodium nitroprusside-induced endothelium-independent relaxation, and thrombin-mediated vasorelaxation did not differ between control and connector saphenous vein grafts at either time point studied. Twenty-seven patients received St Jude Medical connectors. There was no hospital mortality; patients were followed for 679 +/- 241 days. There was 1 late death; the connector saphenous vein graft was patent at postmortem. All connector saphenous vein grafts were patent at follow-up angiography. Four grafts had stenoses (30%-60%), without symptoms or requirement for intervention. All hand-sewn saphenous vein grafts were also patent. CONCLUSIONS: The St Jude Medical connector does not impair endothelium-dependent vasorelaxation. In patients, patency of the connector saphenous vein grafts 6 to 12 months postoperatively was 100% but 22% of grafts had non-flow-limiting stenoses at or near the connector. Further long-term studies are required to confirm the safety of the St Jude Medical connector with regards to endothelial function and restenosis.
Assuntos
Ponte de Artéria Coronária/instrumentação , Veia Safena/transplante , Grau de Desobstrução Vascular/fisiologia , Anastomose Cirúrgica/instrumentação , Anastomose Cirúrgica/métodos , Estudos de Casos e Controles , Angiografia Coronária , Ponte de Artéria Coronária/métodos , Doença das Coronárias/diagnóstico por imagem , Doença das Coronárias/cirurgia , Vasos Coronários/patologia , Vasos Coronários/fisiologia , Endotélio Vascular/fisiologia , Feminino , Seguimentos , Oclusão de Enxerto Vascular/epidemiologia , Rejeição de Enxerto , Sobrevivência de Enxerto , Humanos , Masculino , Nitroprussiato/farmacologia , Veia Safena/cirurgia , Sensibilidade e Especificidade , Técnicas de Sutura , Coleta de Tecidos e Órgãos , VasodilataçãoRESUMO
OBJECTIVES: Evidence has accrued to suggest that diabetic patients face an increased risk of ischemic events and low output syndrome and might mount an inordinate response to ischemia and reperfusion. Because hyperglycemia is a potent stimulus for endothelin-1 production, we hypothesized that increased production, action, or both of endothelin-1 in diabetes might represent an important mediator of endothelial dysfunction in patients with that disease. To this aim, we compared the effects of cardioplegic arrest and reperfusion on coronary sinus effluent endothelin-1 levels and atrial arteriolar vascular responses in diabetic and case-matched nondiabetic patients undergoing coronary artery bypass grafting. METHODS: In study 1 coronary sinus effluent endothelin-1 levels were assessed at baseline and at 1 and 10 minutes after reperfusion in 13 diabetic and 12 nondiabetic patients matched for age, ejection fraction, Parsonnet score, and crossclamp time. In study 2 vascular responses of atrial arterioles subjected to perioperative ischemia-reperfusion were evaluated with videomicroscopy. Atrial microvessels (from appendages) were obtained before and after removal of the aortic crossclamp, and vascular responses to exogenously administered endothelin-1 (10(-10) mol/L) and substance P (10(-8) mol/L) were studied in the presence or absence of BQ-123, an endothelin A receptor antagonist. RESULTS: Diabetic patients elaborated more endothelin-1 at 1 and 10 minutes after reperfusion (P =.01). Endothelin-1-mediated vasoconstriction was similar in diabetic and nondiabetic atrial microvessels before cardioplegic arrest and cardiopulmonary bypass. After cardiopulmonary bypass and reperfusion, endothelin-1-mediated vasoconstriction was enhanced in both groups; however, this response was greater in microvessels from diabetic patients (P =.02). BQ-123, the endothelin A antagonist, attenuated the effects of bypass and reperfusion on endothelin-1-mediated vasoconstriction in both groups (P =.01). Substance P-mediated vasodilatation was similar in diabetic and nondiabetic atrial microvessels before bypass. After bypass and reperfusion, substance P-mediated vasodilatation was diminished in both groups; however, this response was more pronounced in the diabetic group (P =.003). BQ-123 coincubation restored substance P-mediated vasodilatation in both groups. CONCLUSIONS: We determined the following: (1) the coronary effluent release of endothelin-1 is higher in diabetic than in nondiabetic patients after cardiopulmonary bypass and reperfusion; (2) diabetic coronary microvessels respond to bypass and reperfusion with greater endothelin-1-mediated vasoconstriction and diminished nitric oxide-mediated vasodilatation; and (3) these effects are attenuated by endothelin antagonism. Endothelin-1 might be an important mediator of ischemia-reperfusion injury in patients with diabetes. Furthermore, use of endothelin receptor antagonists might be a novel strategy for improving the resistance of the diabetic heart to cardioplegic arrest and reperfusion.
Assuntos
Ponte de Artéria Coronária , Vasos Coronários/fisiopatologia , Angiopatias Diabéticas/fisiopatologia , Endotelina-1/sangue , Endotélio Vascular/fisiopatologia , Idoso , Antagonistas dos Receptores de Endotelina , Parada Cardíaca Induzida , Humanos , Pessoa de Meia-Idade , Peptídeos Cíclicos/farmacologia , Período Pós-OperatórioRESUMO
OBJECTIVES: We have previously demonstrated an importance of endothelin-1 in diabetic patients undergoing bypass surgery. Recent evidence suggests that cardiomyocytes might also produce endothelin-1, which might directly impair myocyte contractility by increasing intracellular calcium levels. Because hyperglycemia is a potent stimulus of endothelin-1 production, we hypothesized that increased production, action, or both of endothelin-1 might be a mediator of direct cardiomyocyte injury in diabetes. Therefore we studied the effects of endothelin receptor blockers (BQ-123 and bosentan) on hyperglycemia-induced endothelin-1 production and cellular injury after ischemia-reperfusion. METHODS: Using a human ventricular heart cell model of simulated ischemia-reperfusion, we studied the effects of normoglycemia (5 mmol/L, 48 hours) and hyperglycemia (25 mmol/L, 48 hours) on cellular injury and endothelin-1 production. Furthermore, the effects of selective endothelin-A and mixed endothelin-A/B receptor antagonism (with BQ-123 and bosentan, respectively) were evaluated. RESULTS: Cellular injury, as assessed by means of trypan blue uptake, was higher in human ventricular heart cells subjected to hyperglycemia and simulated ischemia-reperfusion injury (P =.01); this effect was prevented with both BQ-123 and bosentan (P =.01). In addition, heart cells from the hyperglycemic group elaborated more endothelin-1 after ischemia-reperfusion (P =.02). CONCLUSIONS: Endothelin-1 production and cellular injury were greater in human ventricular heart cells subjected to hyperglycemic conditions and simulated ischemia-reperfusion. These effects are mediated by endothelin-A receptors because both BQ-123 and bosentan exerted similar degrees of protection. Endothelin receptor blockade is a novel strategy to improve the resistance of the diabetic heart to cardioplegic arrest and reperfusion.
Assuntos
Anti-Hipertensivos/farmacologia , Angiopatias Diabéticas/fisiopatologia , Antagonistas dos Receptores de Endotelina , Endotelina-1/sangue , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Peptídeos Cíclicos/farmacologia , Sulfonamidas/farmacologia , Bosentana , Células Cultivadas , HumanosRESUMO
BACKGROUND: Angiotensin II and endothelin-1 are potent endothelium-derived contracting factors. The effects of acute endothelin antagonism on endothelial function in saphenous vein from patients treated with and without angiotensin-converting enzyme inhibitors were compared. METHODS: Vascular segments of saphenous vein were obtained perioperatively from 14 patients on angiotensin-converting enzyme inhibitors and 29 controls. In vitro endothelium-dependent and -independent responses to acetylcholine and sodium nitroprusside were assessed by constructing isometric dose-response curves in precontracted rings in the presence and absence of bosentan (endothelinA/B receptor antagonist) and BQ-123 (endothelinA antagonist) using isolated organ baths. Percent maximum relaxation and sensitivity were compared between interventions. RESULTS: Endothelium-dependent relaxation to acetylcholine was augmented in the angiotensin-converting enzyme inhibitor-treated group (p < 0.005). Both specific and mixed endothelin receptor blockade improved acetylcholine-mediated relaxation in the angiotensin-converting enzyme inhibitor-treated and untreated groups (p < 0.02). The effects of these antagonists were endothelium specific as endothelium-independent responses to sodium nitroprusside remain unaltered. CONCLUSIONS: These data demonstrate that (1) chronic angiotensin-converting enzyme inhibition improves endothelial function in saphenous veins, and (2) this effect can be further augmented by acute endothelin blockade. These data suggest that antagonism of both angiotensin II and endothelin may be important in attenuating saphenous vein arteriosclerosis.
Assuntos
Inibidores da Enzima Conversora de Angiotensina/farmacologia , Antagonistas dos Receptores de Endotelina , Endotélio Vascular/fisiologia , Veia Safena/fisiologia , Vasodilatação/fisiologia , Acetilcolina/farmacologia , Angiotensina II/fisiologia , Bosentana , Ponte de Artéria Coronária , Relação Dose-Resposta a Droga , Endotelina-1/fisiologia , Endotélio Vascular/efeitos dos fármacos , Humanos , Técnicas In Vitro , Pessoa de Meia-Idade , Nitroprussiato/farmacologia , Peptídeos Cíclicos/farmacologia , Veia Safena/efeitos dos fármacos , Sulfonamidas/farmacologia , Vasoconstrição/fisiologia , Vasodilatação/efeitos dos fármacos , Vasodilatadores/farmacologiaRESUMO
PURPOSE: To quantify daily bladder size and position variations during bladder cancer radiotherapy. METHODS AND MATERIALS: Ten bladder cancer patients underwent daily cone beam CT (CBCT) imaging of the bladder during radiotherapy. Bladder and planning target volumes (bladder/PTV) from CBCT and planning CT scans were compared with respect to bladder center-of-mass shifts in the x (lateral), y (anterior-posterior), and z (superior-inferior) coordinates, bladder/PTV size, bladder/PTV margin positions, overlapping areas, and mutually exclusive regions. RESULTS: A total of 262 CBCT images were obtained from 10 bladder cancer patients. Bladder center of mass shifted most in the y coordinate (mean, -0.32 cm). The anterior bladder wall shifted the most (mean, -0.58 cm). Mean ratios of CBCT-derived bladder and PTV volumes to planning CT-derived counterparts were 0.83 and 0.88. The mean CBCT-derived bladder volume (+/- standard deviation [SD]) outside the planning CT counterpart was 29.24 cm(3) (SD, 29.71 cm(3)). The mean planning CT-derived bladder volume outside the CBCT counterpart was 47.74 cm(3) (SD, 21.64 cm(3)). The mean CBCT PTV outside the planning CT-derived PTV was 47.35 cm(3) (SD, 36.51 cm(3)). The mean planning CT-derived PTV outside the CBCT-derived PTV was 93.16 cm(3) (SD, 50.21). The mean CBCT-derived bladder volume outside the planning PTV was 2.41 cm(3) (SD, 3.97 cm(3)). CBCT bladder/ PTV volumes significantly differed from planning CT counterparts (p = 0.047). CONCLUSIONS: Significant variations in bladder and PTV volume and position occurred in patients in this trial.
Assuntos
Tomografia Computadorizada de Feixe Cônico , Neoplasias da Bexiga Urinária/diagnóstico por imagem , Neoplasias da Bexiga Urinária/radioterapia , Bexiga Urinária/diagnóstico por imagem , Idoso , Idoso de 80 Anos ou mais , Fracionamento da Dose de Radiação , Feminino , Humanos , Masculino , Movimento , Tamanho do Órgão , Radioterapia Assistida por Computador/métodos , Análise de RegressãoRESUMO
Protective effects of L-arginine were evaluated in a human ventricular heart cell model of low-volume anoxia and reoxygenation independent of alternate cell types. Cell cultures were subjected to 90 min of low-volume anoxia and 30 min of reoxygenation. L-Arginine (0-5.0 mM) was administered during the preanoxic period or the reoxygenation phase. Nitric oxide (NO) production, NO synthase (NOS) activity, cGMP levels, and cellular injury were assessed. To evaluate the effects of the L-arginine on cell signaling, the effects of the NOS antagonist N(G)-nitro-L-arginine methyl ester, NO donor S-nitroso-N-acetyl-penicillamine, guanylate cyclase inhibitor methylene blue, cGMP analog 8-bromo-cGMP, and ATP-sensitive K+ channel antagonist glibenclamide were examined. Our data indicate that low-volume anoxia and reoxygenation increased NOS activity and facilitated the conversion of L-arginine to NO, which provided protection against cellular injury in a dose-dependent fashion. In addition, L-arginine cardioprotection was achieved by the activation of guanylate cyclase, leading to increased cGMP levels in human heart cells. This action involves a glibenclamide-sensitive, NO-cGMP-dependent pathway.