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Biomedical subjects

B A Keagy

Publications and source records attributed to B A Keagy.

At least 19 recordsLinked to original sources
PubMed1992

Flow cytometric analysis of organ preservation-induced endothelial cell membrane damage.

Organ preservation for transplantation is associated with endothelial cell damage. This vascular injury results in increased capillary permeability, graft edema, and early graft dysfunction. This damage may be the limiting factor in preservation of these organs. This study uses flow cytometric assessment of membrane integrity to examine the effects of various organ preservation solutions on human umbilical vein endothelial cell cultures. Confluent plates of human umbilical vein endothelial cells were incubated at 4 degrees C for 24, 48, and 72 hours in commonly used preservation solutions. After cold incubation, the cells were harvested and stained with propridium iodide and fluorescein diacetate. Cells were examined using a flow cytometer for membrane integrity and cytosolic activity. When examined after 24, 48, and 72 hours, cells stored at 4 degrees C in a 5% polyethylene glycol salt solution were significantly less damaged than those stored in any other solution (p less than 0.05). After 48 and 72 hours at 4 degrees C, cells stored in ViaSpan were significantly more intact than cells stored in EuroCollins and 0.9% saline solution (p less than 0.05). This study demonstrates that endothelial cell damage occurs during cold storage and that a polyethylene glycol-based solution showed superior cellular preservation.

Adenosine↗
PubMed1992

Isolated lung transplantation for end-stage lung disease: a viable therapy.

Since January 1990, we have performed 29 isolated lung transplantations in 28 patients with end-stage lung disease (12 single, 16 bilateral). Recipient diagnoses were: cystic fibrosis (11), chronic obstructive pulmonary disease (6), pulmonary fibrosis (6), eosinophilic granulomatosis (1), postinfectious lung disease (1), adult respiratory distress syndrome (1), and primary pulmonary hypertension (2). There have been four deaths, two in patients with pulmonary fibrosis and two in patients with primary pulmonary hypertension. Four patients have undergone transplantation while on ventilatory support for respiratory failure (2 with cystic fibrosis, 1 having redo lung transplantation with cystic fibrosis, and 1 with adult respiratory distress syndrome); all of these have survived. Six patients required cardiopulmonary bypass, which was associated with increased transfusion requirement. All patients 2 months after discharge have returned to an active life-style, except for 2 patients who currently await retransplantation. Preoperative pulmonary rehabilitation has resulted in significant improvement in exercise performance in all patients. Immunosuppression consists of cyclosporine, azathioprine, and antilymphoblast globulin (University of Minnesota), withholding systemic steroids in the early postoperative period. We have employed bronchial omentopexy in all but four transplants; there has been one partial bronchial dehiscence, two instances of bronchomalacia requiring internal stenting, and one airway stenosis. Cytomegalovirus disease has been seen frequently (15 cases), but has responded well to treatment with ganciclovir. Other complication shave included one drug-related prolonged postoperative ventilation, thrombosis of a left lung after bilateral lung transplantation requiring retransplantation, five episodes of unilateral phrenic nerve palsy after bilateral lung transplantation (4 resolved), and the requirement of massive transfusion (greater than 10 units) in 5 patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1992

Descending thoracic aorta-to-iliofemoral artery bypass as an alternative to aortoiliac reconstruction.

During the last 3 decades subcutaneous extraanatomic bypass, despite its limited durability, has been the favored alternative to infrarenal aortofemoral bypass. Meanwhile, the descending thoracic aorta has been scarcely used as an inflow source for aortoiliac reconstruction. Over the past 8 years we performed 16 bypasses from the descending thoracic aorta to the iliofemoral vessels for occlusive disease. Our experience combined with that found in the English-language literature totaled 141 patients. In 79 patients (56%) the indication for surgery was failure or infection of an abdominal aortofemoral graft. Previous abdominal operations, sepsis, radiation therapy, the presence of abdominal stomas, or an unsuitable infrarenal aorta were the indications in the remaining cases. The combined operative mortality rate was 6.4%. The life-table primary graft patency was 98% at 1 year, 88% at 2 years, and 70.4% at 5 years. Bypass from the descending thoracic aorta to the iliofemoral artery uses an inflow source superior to other extraanatomic reconstructions, does not require aortic cross-clamping, avoids the abdominal cavity, and places the graft remote to the skin and intestine. The operative mortality and patency rates compare favorably to those of other extraanatomic or remedial aortic reconstructions. Descending thoracic aorta to iliofemoral artery bypass is a superb alternative to abdominal aortofemoral bypass, carries a low mortality rate, has an excellent short-term patency, and features unique characteristics for long-term durability.

Adult↗
PubMed1992

Single lung transplantation for eosinophilic granulomatosis.

A 26-year-old woman with end-stage lung disease due to eosinophilic granulomatosis had single right lung transplantation with an excellent function result that persists beyond 9 months of follow-up. Single lung transplantation offers excellent palliation to selected patients with end-stage lung disease.

Adult↗
PubMed1992

Endothelial cell preservation using organ storage solutions.

The development of organ preservation solutions has been primarily accomplished in whole organ animal models. This study examines the toxicity of commonly used organ preservation solutions on endothelial cells in vitro. Primary human umbilical-vein endothelial cell cultures were incubated at 4 degrees C in solutions of 0.9% saline (NS), EuroCollins, ViaSpan (Belzer UW) (VIA), or Hank's balanced salts with 5% polyethylene glycol (PEG). Endothelial cell viability was ascertained by colormetric measurement of mitochondrial reduction of 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to purple 1-(4,5-Dimethylthiazol-2-yl)-3,5-diphenylformazan. After exposure to hypothermic storage, cells were incubated at 37 degrees C in media with MTT, and the amount of reduced formazan present was quantified using a micro-ELISA spectrophotometer. Higher absorbance values were indicative of better cellular preservation. Cells stored in PEG displayed the highest viability at all time periods. ViaSpan provided better cellular protection than EC at longer storage intervals. This model allows for rapid assessment of preservation-induced endothelial cell injury and may aid development of improved storage techniques.

Absorption↗
PubMed1992

Improvement of endothelial cell viability at 4 degrees C by addition of lazaroid U74500A to preservation solutions.

Lazaroids are potent inhibitors of lipid peroxidation. Endothelial cell damage has been shown to occur during cold storage preservation of lung and liver. This study examines the effects on endothelial cell viability of the addition of four lazaroids, U74006F, U78518F, U74500A, or U75412E to preservation solutions. Human umbilical vein endothelial cell cultures were stored at 4 degrees C for 48 and 96 hr in EuroCollins or 5% polyethylene glycol in buffered saline (PEG). U78518F, U74500A, U74006F, U75412E, or dexamethasone (each 50 microM) was added to EC (n = 32) or PEG (n = 32) and compared with control solutions of EC or PEG alone. Endothelial cell viability was determined by measuring cellular reduction of 3-[4,5-dimethylthiazol-2-yl]-2,3-diphenyltetrazolium bromide to a purple formazan dye. The reduction occurs only in viable cells and requires mitochondrial dehydrogenase activity. Results were quantified by measuring dye absorbance (Ab) with a micro-ELISA spectrophotometer. Absorbance values were compared by ANOVA and reported as mean values +/- standard deviation. Addition of U74500A to EC (Ab = 0.474 +/- 0.055) and PEG (Ab = 0.462 +/- .005) improved viability at 48 hr when compared with EC (Ab = 0.289 +/- 0.069) and PEG (Ab = 0.287 +/- 0.052) alone (P less than 0.05). At 96 hr, addition of U74500A resulted in improved viability in both EC (Ab = 0.377 +/- 0.068) and PEG (Ab = 0.195 +/- 0.09) or PEG alone (Ab = 0.212 +/- 0.1) (P less than 0.05). Other lazaroids tested were also effective in improving cellular viability, but to a lesser degree than U74500A. This study demonstrates that the addition of lazaroids to organ preservation solutions improves endothelial cell viability.

Absorption↗
PubMed1992

The course of severe foot infection in patients with diabetes.

Infection of the foot is a limb threatening condition for patients with diabetes mellitus. Identification of patients with diabetes and severe infection of the foot most likely to benefit from early revascularization or major amputation would improve the results of a treatment policy to prevent limb loss and avoid futile delays in amputation. During a nine year period, 79 diabetic patients underwent emergency procedures for severe infection of the foot during the initial hospitalization period. None of the patients underwent arterial reconstruction. Eventually, 21 of the patients required a major amputation, eight during the initial hospitalization and 13 on a subsequent admission. Stepwise discriminant analysis of clinical independent variables revealed that the patients most likely to require a major amputation during the initial hospitalization were those with an absent dorsalis pedis pulse and a polymicrobial infection (p = 0.018). The overall amputation rate (immediate or subsequent amputation) was higher for patients of either sex with nonpalpable pedal pulses when compared with those with at least one palpable pulse (p less than 0.05). Males who were not dependent on insulin had the highest risk of overall limb loss (p = 0.01). Patients undergoing delayed amputation required a significantly higher number (p = 0.01) of readmissions to the hospital for recurrent infection of the foot than those who did not undergo amputation. Data in the current study suggest that early major amputation in a subset of patients would prevent delay in the rehabilitation process of the amputee, decrease long term morbidity and reduce health care cost. An aggressive policy of early revascularization in patients with a pulse deficit may reduce the amputation rate in those with diabetes with severe infection of the foot.

Amputation, Surgical↗
PubMed1991

Enhanced pulmonary function using dimethylthiourea for twelve-hour lung preservation.

Current preservation techniques for lung transplantation limit ischemic time to 6 hours. The purpose of this study was to evaluate the ability of dimethylthiourea, a low molecular weight free radical scavenger, to prolong this interval. An in vivo canine transplantation model was used to assess lung function. At harvest and after circulatory arrest, the donor lung was flushed with modified Euro-Collins solution (50 mL/kg). In a blinded fashion, dimethylthiourea (5 g) or saline solution was added to the flush solution at harvest and also infused (20 g over 2 hours) at reimplantation. Harvested lungs were stored for 12 hours at 4 degrees C. Allotransplantation was performed in recipient dogs ventilated with 40% O2. After 1 hour, the contralateral pulmonary artery was ligated, forcing the dog to be dependent on the transplanted lung. Twelve dogs were studied, with 6 randomly assigned to each treatment group in a blinded fashion. Measurements were recorded for 8 hours, keeping the inspired oxygen fraction constant at 0.40. All dimethylthiourea-treated dogs survived the observation period, whereas one third of the dogs that received saline solution died. Dimethylthiourea-treated dogs had significantly greater arterial oxygen tension and significantly less pulmonary vascular resistance compared with control animals. These results suggest that treatment of the lung with a free radical scavenger (dimethylthiourea) improves pulmonary function after reimplantation in a canine model after 12-hour hypothermic storage.

Animals↗
PubMed1991

A strategy to increase the donor pool: use of cadaver lungs for transplantation.

A shortage of suitable donors is a serious obstacle to the widespread application of isolated lung transplantation for end-stage lung disease. We hypothesized that lung tissue likely remains viable for a sufficient period of time to allow for safe postmortem retrieval of lungs for transplantation. Studies were conducted in a nonsurvival model of canine lung allotransplantation. Donor animals were sacrificed, and subsequent lung harvest was delayed for 1 hour, 2 hours, or 4 hours. Pulmonary retrieval was then performed in a standard fashion, flushing the lung block with modified Euro-Collins solution. Lungs were then stored for 4 hours before single allotransplantation. Recipient animals were maintained anesthetized, and followed up for 8 hours. By occlusion of the pulmonary artery and bronchus to the native lung, recipient animals were forced to survive solely on the transplanted lung, with a constant inspired oxygen fraction of 0.40. All 5 recipient animals of 1-hour cadaver lungs survived the 8-hour observation period with excellent hemodynamics and gas exchange. Two of 5 recipients of 2-hour cadaver lungs survived the observation period, whereas a third animal survived for 5 hours with excellent gas exchange. One of 4 animals transplanted with a 4-hour cadaver lung survived the observation period. Retrieval of lungs from cadavers whose hearts are not beating may prove to be a safe and effective method to increase the pulmonary donor pool.

Animals↗
PubMed1991

Intermittent claudication.

The natural history of intermittent claudication is generally benign. Clear indications for revascularization in patients with intermittent claudication are rest pain and necrotic tissue. Disabling claudication in patients who are at low operative risk is another acceptable indication for surgical treatment. Young patients with intermittent claudication from aortoiliac occlusion constitute a subset of patients in whom a more aggressive approach is justified. Nevertheless, in most patients, the management of intermittent claudication should be conservative.

Female↗
PubMed1990

Objective measurement of limb perfusion by dermal fluorometry. A criterion for healing of below-knee amputation.

Quantitative fluorometry has been recommended as an accurate adjunct to clinical judgment in the preoperative assessment of lower-extremity amputation level. In this prospective study of 56 patients who had below-knee amputation, clinical judgment was used as the sole criterion for site selection. Quantitative fluorometry was compared with clinical judgment in a prospective, blinded study. All patients were studied before amputation with administration of intravenous fluorescein. Fifteen minutes after injection, objective measurement of dye fluorescence was performed at multiple sites with a quantitative fluorometer, and a dye fluorescence index was derived. All limbs undergoing amputation were ischemic, manifested by rest pain, nonhealing ulcers, or gangrene. Five patients (8.7%) failed to heal at the below-knee level. The mean dye fluorescence index for the group that healed was 81 +/- 51 (range, 13 to 259) and for the group that failed to heal, 110 +/- 49 (range, 70 to 195). Objective measurement of fluorescein perfusion did not correlate with amputation healing at the below-knee level in our patient population.

Adult↗
PubMed1990

Oxygen free radical scavengers decrease reperfusion injury in lung transplantation.

An in vivo canine model was used to assess the ability of an oxygen free radical scavenger to decrease reperfusion injury in lung transplantation. In 12 dogs, the left lungs were transplanted after they had been preserved for 24 hours at 4 degrees C after pulmonary artery flushing with modified Eurocollins solution. In 6 dogs, dimethylthiourea, a potent oxygen free radical scavenger, was added to the flush solution and was also given to the recipients just before reperfusion. In all animals, the contralateral pulmonary artery and bronchus were ligated and lung function was assessed for 12 hours or until death. Three dogs died prematurely in the control group, whereas only 1 dog died prematurely in the dimethylthiourea group. This resulted in a statistically significant difference in the average length of survival (p less than 0.05). Pulmonary artery and right atrial pressures were significantly lower in the dimethylthiourea group during the first 6 hours (p less than 0.05). Treatment with dimethylthiourea resulted in a significantly higher arterial oxygen tension at 4 hours, and intrapulmonary shunt tended to be lower. Thus, it would appear that dimethylthiourea has a protective effect on lungs preserved for 24 hours before transplantation in dogs.

Animals↗
PubMed1990

Endothelial cell thromboxane production and its inhibition by a calcium-channel blocker.

Vascular endothelium has been established as a major source of prostacyclin production. Whether endothelial cells are also capable of synthesizing thromboxane A2 remains controversial. Suppression of such endothelial thromboxane synthesis would be beneficial for short-term patency of vascular grafts. This study examined the production of thromboxane A2 by endothelial cells and its modulation by the calcium-channel blocker nifedipine in vitro. The results indicate that (1) endothelial cells spontaneously secrete thromboxane A2, (2) this production can be enhanced severalfold in the presence of arachidonic acid, and (3) the calcium-channel blocker nifedipine significantly inhibits thromboxane production without demonstrable toxicity to the endothelial cells.

Animals↗
PubMed1990

Endothelial cells can synthesize leukotriene B4.

Leukotriene B4 (LTB4) from vascular endothelium may play a key role in the genesis of atherosclerotic lesions. However, the ability of this tissue to synthesize LTB4 is controversial. To resolve this issue arachidonic acid metabolism was characterized in cultures of confluent monolayers of a rabbit aortic endothelial cell line by use of both high-pressure liquid chromatography and radioimmunoassay. Cells were grown to confluence in Dulbecco's modified Eagle's medium/Ham's F12 with 5% fetal bovine serum. Lipoxygenase activity was studied by placing the cells in Hank's balanced salt solution with 2 mumol/L indomethacin. After 30 minutes preincubation with indomethacin cells were exposed to either arachidonic acid (10 mumol/L) or arachidonic acid labeled with radioactive carbon (14C) (1 microCi; SA 58 mCi/mmol) and then stimulated with 9.5 mumol/L calcium ionophore A23187 for 55 minutes. Studies of the cyclooxygenase activity were performed without preincubating with indomethacin. Samples were prepared for high-pressure liquid chromatography by evaporation to dryness under a vacuum and resuspending in 2 ml of 1:1 methanol/water. Tritium-labeled standards were added before loading the 14C-labeled samples on the column. Radiolabeled arachidonic acid metabolites were separated by high-pressure liquid chromatography and detected by means of a dual channel flow-through radiodetector that monitors both 14C and 3H. Based on coelution with authentic standards three lipoxygenase metabolites of arachidonic acid have been identified: LTB4, 12- and 5-hydroxyeicosatetraneoic acid. Leukotriene B4 was further characterized by ultraviolet spectral analysis and inhibition studies with use of nordihydroguaiaretic acid. Quantitation was facilitated by commercially available radioimmunoassay kits. An average of 600 pg LTB4/10(6) cells was measured from separate experiments.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Reperfusion injury in the lung preserved for 24 hours.

The left lower lobes of 28 canine lungs were isolated, preserved, and then reperfused for 150 minutes. Five groups of lobes were studied: group 1, control (n = 5); group 2, one hour of warm ischemia (n = 5); group 3, one hour of warm ischemia + oxygen free radical scavengers (n = 5); group 4, 24 hours of cold ischemia (n = 8); and group 5, 24 hours of cold ischemia + oxygen free radical scavengers (n = 5). Oxygen free radical scavengers consisted of superoxide dismutase and catalase (100 micrograms/mL) given at the moment of reflow. Extravascular lung water (grams per gram of blood-free dry lobe weight) after reperfusion was 2.75 +/- 0.19, 5.46 +/- 0.60, 4.08 +/- 0.37, 9.43 +/- 0.98, and 6.91 +/- 0.95 for groups 1 through 5, respectively (p less than 0.05, groups 2 through 5 versus group 1; p less than 0.05, group 2 versus group 3 and group 4 versus group 5). Lung tissue lipid peroxidation, measured as thiobarbituric acid reactive material, was 117 +/- 14, 314 +/- 19, and 163 +/- 25 nmol/g dry lobe weight for groups 1, 4, and 5, respectively (p less than 0.05, group 4 versus group 1 and group 4 versus group 5). The data suggest that oxygen free radical scavengers attenuate reperfusion injury after long-term hypothermic lung preservation.

Animals↗
PubMed1989

Improved lung preservation using a dimethylthiourea flush.

The purpose of this study was to evaluate the ability of dimethylthiourea (DMTU), a low molecular weight hydroxyl free radical scavenger, to improve preservation of the lung for transplantation. Following preservation, 15 isolated canine left lower lobes were reperfused for 90 min with autologous blood. Five group I lobes served as controls and were not subjected to ischemia prior to reperfusion. Five group II lobes were flushed and submerged in a cold Euro-Collins solution and stored for 4 hr at 4 degrees C prior to reperfusion. Group III lobes were flushed with a 20 mM DMTU-enhanced Euro-Collins solution, stored for 4 hr, and then reperfused. The isogravimetric method was utilized to determine the capillary permeability coefficient (Kfc) for the reperfused lobes. The Kfc values were 0.10 +/- 0.01, 0.17 +/- 0.01, and 0.10 +/- 0.008 ml/min/mm Hg/100 g lung for groups I, II, and III, respectively (P less than 0.01 II vs I, III). Extravascular lung water values in the reperfused lobe were 4.44 +/- 0.45, 6.57 +/- 0.38, and 5.23 +/- 0.22 ml/g blood free dry lung weight for groups I, II, and III (P less than .05, II vs. I, III). Lung lipid peroxidation, measured as thiobarbituric acid-reactive material, was higher in group II, 146 +/- 6 nmole/g, than in either group I, 90 +/- 5 nmole/g, or group III, 91 +/- 4 nmole/g (P less than 0.01). The results indicate that the addition of DMTU improves hypothermic lung preservation by reducing lipid peroxidation and edema formation upon reperfusion.

Animals↗
PubMed1988

Pulmonary blood velocity profile variability in open-chest dogs: influence of acutely altered hemodynamic states on profiles, and influence of profiles on the accuracy of techniques for cardiac output determination.

Clinical investigations focused on finding characteristics of noninvasively obtained measurements of pulmonary blood velocity that can be used to quantitate pulmonary blood flow and/or pulmonary pressure have often yielded results whose imprecision has been attributed to flow pattern variability. To determine flow pattern variability in an in vivo animal model in varying hemodynamic states, main pulmonary artery blood velocity waveforms were recorded in 17 dogs at 2-mm intervals along an anterior to posterior wall-oriented axis using a 20-MHz pulsed Doppler needle probe. Control data were obtained before the animals were subjected to altered flow (atrial level shunts) and pressure (10% O2 inhalation) states. Instantaneous velocity profiles were computed throughout the cardiac cycle. Estimates of pulmonary blood flow were obtained assuming an elliptical model of the pulmonary artery which allowed computation of velocity at all points in the cross section, based on the measured values along the axis. Model-based estimates were compared to measured values and estimates obtained in the traditional fashion, i.e., the product of centerline velocity and cross-sectional area. Results clearly showed marked interanimal variability, even in control states. Reverse flow in the posterior half of the vessel, which tended to become more pronounced with increased pulmonary artery pressure, was observed during late systole and early diastole. Elevated pulmonary blood flow tended to increase the maximum velocities along the anterior wall relative to midline velocities. Neither estimate of cardiac output yielded consistently accurate results (r = 0.77 for model-based method, r = 0.80 for area times central velocity method). Findings of this study, which highlight the dependency of waveform characteristics on sampling site, the large degree of intersubject variability, and the need for large or multiple sample volumes for pulmonary blood flow determination, help clarify inconsistencies observed by clinicians and suggest that future work with animal models will facilitate a greater understanding of the determinants of human pulmonary velocity waveforms.

Animals↗