Biomedical subjects

C G Wheeler

Publications and source records attributed to C G Wheeler.

12 recordsLinked to original sources

PubMed2001

Ethanol-induced up-regulation of the urokinase receptor in cultured human endothelial cells.

BACKGROUND: Moderate alcohol consumption has been correlated to reduced coronary artery disease (CAD) risk and mortality. This alcohol effect may be mediated in part by an increased endothelial cell (EC) fibrinolysis. ECs synthesize fibrinolytic proteins, tissue plasminogen activator (t-PA), urokinase type plasminogen activator (u-PA), and plasminogen activator inhibitor type-1(PAI-1). In addition, they synthesize and regulate receptors for fibrinolytic proteins, namely (t-PA and plasminogen receptor) Annexin II and u-PA receptor (u-PAR). These receptors play an important role in the regulated expression of receptor-bound plasminogen activator conversion of receptor-bound plasminogen to receptor-bound plasmin on the EC surface (surface-localized fibrinolytic activity). Therefore, systemic factors, such as ethanol, that affect the level, or activity or interaction of one or more of these components, resulting in the increased expression of surface-localized EC fibrinolytic activity, will be expected to reduce the risk for thrombosis, CAD, and myocardial infarction (MI). We have previously shown that low ethanol up-regulates t-PA and u-PA gene transcription, while it down-regulates PAI-1, hence resulting in increased (sustained, 24 hr) surface-localized EC fibrinolytic activity. The current studies were carried out to determine whether low ethanol increased u-PAR expression in cultured human umbilical cord vein ECs (HUVECs). METHODS: Cultured HUVECs were preincubated (1 hr) in the absence/presence of ethanol (0.025-0.2%, v/v); u-PAR mRNA (RT-PCR), antigen (western blot), and activity (125I-u-PA ligand binding/Scatchard analysis) levels were then measured after 0-24 hr. To determine whether the ethanol-induced changes in the u-PAR expression were transcriptional, transient transfection studies were carried out using a u-PAR/ luciferase promoter construct (pu-PAR120/luc [1.2-kb u-PAR promoter fragment ligated to a promoterless luciferase vector]). RESULTS: uPAR mRNA levels increased 2- to 3-fold and antigen levels (western blot) increased 2- to 4-fold while u-PA binding activity increased 36% (1.25 vs. 1.7 x 10(5) sites/cell, Bmax) without significantly affecting the Kd (1-2 nM). Transient transfection of cultured HUVECs with a pu-PAR120/luc construct resulted in a 2- to 3-fold increase in promoter activity in ethanol-induced cultures, compared with controls. CONCLUSION: These combined results demonstrate that low ethanol (< or =0.1%, v/v) induces the up-regulation of u-PAR gene transcription, resulting in increased u-PAR ligand binding activity. These results also further identify/define the contribution and role of another fibrinolytic protein in the overall ethanol-induced increase in surface-localized EC fibrinolysis that may underlie and contribute, in part, to the cardioprotection attributed to moderate alcohol consumption.

Cells, Cultured↗

PubMed2001

Ethanol-induced increased surface-localized fibrinolytic activity in cultured human endothelial cells: kinetic analysis.

BACKGROUND: Moderate alcohol consumption is associated with reduced risk for coronary heart disease and this cardioprotection may be due, in part, to increased fibrinolysis. We have previously demonstrated that low concentrations of ethanol (0.1%, v/v) induce the short-term (<1 hr) and sustained, long-term (24 hr) increase in surface-localized fibrinolytic activity; it up-regulates t-PA, u-PA, and the candidate plasminogen receptor (PmgR), annexin II, and gene transcription in cultured human umbilical vein endothelial cells (HUVECs). These studies describe the short- and long-term effects of low concentrations of ethanol on the kinetics of cell-bound 125I-labeled Glu-plasminogen (Glu-Pmg) activation by receptor (R)-bound t-PA, resulting in increased fibrinolytic activity in cultured HUVECs. METHODS: Live cultured HUVECs were incubated with varying concentrations of Glu-Pmg (0.25-2 /M) and ethanol (0.025-0.1%, v/v) (in the presence of Aprotinin and alpha2-antiplasmin) and the direct activation of cell-bound 125I-labeled Glu-Pmg quantitated by measurement of 125I-labeled Mr 20 kDa plasmin light-chain, after reduction/SDS-PAGE. The effects of ethanol on '25I-labeled Glu-Pmg and t-PA ligand binding were determined by Scatchard analysis (Bmax, sites/cell). RESULTS: Cell-bound t-PA (endogenous/exogenous) activation of cultured HUVEC-bound 125I-labeled Glu-Pmg (short- and long-term) obeyed Michaelis-Menten kinetics, both in the absence/presence of low ethanol, as shown by Lineweaver-Burke plot analysis. In the short-term, ethanol (at 0.1%) increased the Vmax (2.5-fold), kcat (2-fold) and the apparent kcat/Km (4-fold), commensurate with a significant decrease in the apparent Km (6-fold) and increase in '25I-labeled Glu-Pmg ligand binding, Bmax (2-fold). In the long-term, ethanol increased the Vmax (2- to 3-fold), kcat (2.5-fold), apparent kcat/Km (5-fold), and Bmax (2-fold) for 125I-labeled Glu-Pmg ligand binding, without significantly affecting the apparent K . CONCLUSIONS: Low concentrations of ethanol induce the short- versus long-term increase in surface-localized fibrinolytic activity in cultured HUVECs via different mechanisms. Short-term effects may be mediated by ethanol-induced membrane conformational changes that simultaneously facilitate increased surface-localized HUVEC PmgR availability and fibrinolytic protein/receptor interactions, resulting in the increased affinity of t-PA for Glu-Pmg and the accelerated activation of Glu-Pmg (increased Bmax, decreased apparent Km). The long-term effects may be attributed primarily to the ethanol-induced increased availability of both newly synthesized t-PA and PmgR and, hence, the accelerated activation of Glu-Pmg (increased Bmax)

Cells, Cultured↗

PubMed1999

Amputation prevention by vascular surgery and podiatry collaboration in high-risk diabetic and nondiabetic patients. The Operation Desert Foot experience.

OBJECTIVE: To describe a unique multidisciplinary outpatient intervention for patients at high risk for lower-extremity amputation. RESEARCH DESIGN AND METHODS: Patients with foot ulcers and considered to be high risk for lower-extremity amputation were referred to the High Risk Foot Clinic of Operation Desert Foot at the Carl T. Hayden Veterans Affairs' Medical Center in Phoenix, Arizona, where patients received simultaneous vascular surgery and podiatric triage and treatment. Some 124 patients, consisting of 90 diabetic patients and 34 nondiabetic patients, were initially seen between 1 October 1991 and 30 September 1992 and followed for subsequent rate of lower-extremity amputation. RESULTS: In a mean follow-up period of 55 months (range 3-77), only 18 of 124 patients (15%) required amputation at the level of the thigh or leg. Of the 18 amputees, 17 (94%) had type 2 diabetes. The rate of avoiding limb loss was 86.5% after 3 years and 83% after 5 years or more. Furthermore, of the 15 amputees surviving longer than 2 months, only one (7%) had to undergo amputation of the contralateral limb over the following 12-65 months (mean 35 months). Compared with nondiabetic patients, patients with diabetes had a 7.68 odds ratio for amputation (95% CI 5.63-9.74) (P < 0.01). CONCLUSIONS: A specialized clinic for prevention of lower-extremity amputation is described. Initial and contralateral amputation rates appear to be far lower in this population than in previously published reports for similar populations. Relative to patients without diabetes, patients with diabetes were more than seven times as likely to have a lower-extremity amputation. These data suggest that aggressive collaboration of vascular surgery and podiatry can be effective in preventing lower-extremity amputation in the high-risk population.

Adult↗

PubMed1981

Supraclavicular resection of the first rib for thoracic outlet syndrome.

The experience with supraclavicular transcervical first rib resection (or cervical rib resection) and total scalenotomy reported herein shows this to be a safe and effective treatment for patients with the thoracic outlet syndrome. We recommend this method as a satisfactory anatomic approach for the correction of this condition. Its advantages are as follows: anatomic structures may be visualized fully by both the surgeon and the assistant; complex or recurrent thoracic outlet problems may be dealt with directly; additional procedures (such as vascular graft, neurolysis, neck exploration, sympathectomy and cervical rib resection) may be performed; the procedure can be done in 90 minutes or less; neither the patient nor the assistant is obliged to assume an awkward or strained position and consequently the likelihood of intraoperative iatrogenic injury is reduced; and neither the period of hospitalization nor the period of immobility is longer than with the other operative techniques currently used for this condition.

Adult↗

PubMed1980

Surgical management of total carotid artery occlusion. An update.

Fourteen operations were performed in 13 patients with total carotid artery occlusion. All patients had symptoms ranging in duration from a few hours to several months. Flow wa reestablished in 10 of the 14 arteries operated on, a success rate of 74 percent. Complication included one operative death and one permanent neurologic deficit. An aggressive approach to the surgical management of patients with total carotid artery occlusion is recommended.

Aged↗

PubMed1969