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Biomedical subjects

J C Stanley

Publications and source records attributed to J C Stanley.

At least 145 records · Page 8Linked to original sources

Effect of different oxygen saturations on venous versus arterial endothelial cell growth in vitro.

Canine carotid arterial and external jugular venous endothelial cells were derived, cutured, passed, and pooled. Three X 10(6) cells were seeded into capped roller-bottles. Three roller-bottes were seeded with venous, and another three with arterial derived endothelial cells for each of three "oxygen saturations" 5, 20 and 50% O2 respectively, and incubated for seven days and then counted. Arterial endothelial cells showed no cell proliferation in high oxygen saturation compared to an 83% increase in cell numbers when oxygen supply had been held to 20%. Venous endothelial cells showed no cell proliferation in medium or high oxygen supply but low oxygen supply (5% O2) gave a 91% increase in cell counts. The small number of roller-bottles investigated does not allow firm conclusions but our results warrant further evaluation.

Aerobiosis↗

Treatment of established prosthetic vascular graft infection with antibiotics preferentially concentrated in leukocytes.

The efficacy of treating established vascular graft infections with rifampin and clindamycin (preferentially concentrated in leukocytes) and cefazolin (not concentrated in leukocytes) was studied in a canine model. Infrarenal aortic, 6 mm by 6 cm knitted Dacron double velour grafts were implanted and infected with 10(8) colony-forming units (CFU) of coagulase-positive Staphyloccus aureus organisms injected intravenously immediately after graft placement. Antibiotic therapy was instituted at 3 months postimplantation. Three groups were studied: (I) untreated controls (n = 3); (II) therapy with intravenous cefazolin 15 mg/kg/8 hr for 28 days (n = 7); and (III) combined therapy with intravenous rifampin 13 mg/kg/24 hr and intravenous clindamycin 13 mg/kg/8 hr for 28 days (n = 7). Grafts were removed for quantitative bacteriologic studies after the 28-day course of therapy. Two group I control grafts remained patent with 6.4 X 10(6) and 8.1 X 10(3) CFU S. aureus/gm of graft. The third control graft was thrombosed. Two group II animals demonstrated 1.6 X 10(7) and 2.3 X 10(5) CFU S. aureus organisms/gram of graft, respectively; the remaining five group II grafts were free of organisms. All group III grafts were sterile--a significant difference (p less than 0.05) from group I grafts. In this experimental model, established prosthetic graft infections were eradicated by intensive treatment with antibiotics preferentially concentrated in leukocytes.

Animals↗

Vascular surgery in the United States. Report of the Joint Society for Vascular Surgery--International Society for Cardiovascular Surgery Committee on Vascular Surgical Manpower.

The Joint Committee on Vascular Surgical Manpower was established in 1985 by the Society for Vascular Surgery and the North American Chapter of the International Society for Cardiovascular Surgery. It was charged to provide recommendations regarding vascular surgical manpower requirements for the next 15 years. Analysis of National Center for Health Statistics vascular operative rate data and 1690 questionnaire responses from vascular surgeons documented that vascular surgeons performed 235,400 (41%) of the total of 571,000 vascular operations undertaken in 1985. Vascular surgeons performed 87% of 30,000 aortoiliofemoral reconstructions, 77% of 72,000 peripheral vessel bypasses, 75% of 33,000 abdominal aortic aneurysm repairs, 59% of 55,000 angioaccess procedures, and 50% of 107,000 carotid endarterectomies. However, lack of accurate data on caseloads of surgeons who were not vascular specialists precludes precise prediction of manpower requirements for vascular surgery. It is important that surgical leaders and policy makers define the types of vascular surgical procedures that may be undertaken by vascular and other surgeons. Ongoing analyses must include such determinations to establish accurate data for the prediction of future manpower needs for vascular surgery. Furthermore, future manpower studies should be linked to outcome studies to assess not only numbers of surgeons and operations but quality of care as well.

Forecasting↗

Adrenergic and dopaminergic control of the canine paw microcirculation.

Basal isolated canine paw blood flow was equally distributed between arteriovenous anastomosis (AVA) and capillary circulations. Norepinephrine decreased AVA flow by 92% and capillary flow by 41%. Dopamine significantly reduced AVA flow by 94% compared to baseline with a 37% reduction in capillary flow. However, with alpha-adrenergic blockade dopamine decreased AVA flow 66% while capillary flow increased 42%. Isoproterenol increased capillary flow almost twofold and appeared to decrease AVA flow, although the latter was statistically insignificant. Differential effects of adrenergic and dopaminergic agonists on canine paw AVA and capillary blood flow suggest the existence of independent regulation of these components of the microcirculation.

Animals↗

Nuclide imaging of vascular graft-platelet interactions: comparison of indium excess and technetium subtraction techniques.

Indium-111-labeled platelet adherence to ePTFE thoracoabdominal vascular prostheses in a canine model (n = 10) was quantitated by (1) an indium-111 excess technique, contrasting graft radioactivity to that in a reference region, and (2) a technetium-99m subtraction technique, with radioactivity of circulating platelets eliminated by discounting background blood activity. Variation in graft thrombogenicity was provided by seeding six prostheses with enzymatically derived autologous endothelial cells, and implanting four prostheses without seeding. Grafts were imaged at 1, 4, and 6 weeks postimplantation, with platelet labeling using indium-111-oxine and red blood cell labeling using technetium-99m. At 7 weeks grafts were excised and gamma activity was measured in proximal, middle, and distal segments. Luminal generation of TxB2 and 6-keto-PGF1 alpha from midportions of grafts was assayed. Indium-111 excess ratios at 6 weeks correlated with actual gamma activity of excised grafts (proximal r = 0.80, P less than 0.01; middle r = 0.73, P less than 0.05; distal r = 0.48, ns) but such a correlation did not exist for the technetium-99m subtraction technique (r = -0.05, -0.25, and 0.16, in the three segments, respectively, all ns). The ratio of graft to aortic TxB2 production revealed a positive correlation with graft gamma activity (r = 0.87, P less than 0.01), and the ratio of graft 6-keto-PGF1 alpha to TxB2 production also correlated with gamma counts (r = -0.64, P = 0.05). In this experimental setting technetium-99m subtraction analysis was an imprecise method of detecting graft platelet accumulation, whereas indium-111 excess ratios proved to be a more accurate method of quantitating vascular prosthetic thrombogenicity.

Animals↗

Control of phenylalanine and tyrosine metabolism by phosphorylation mechanisms.

A system for the parallel determination of enzyme phosphorylation and expressed activity in rat liver cells, and its application to studies of phenylalanine hydroxylase and tyrosine aminotransferase, is described. Phenylalanine hydroxylase is phosphorylated by agents which stimulate cyclic AMP- and Ca2+-dependent protein kinase activity. The phosphorylation site(s) appear to be the same for both kinases. Phosphorylation is accompanied by increased metabolic flux at low, physiologically relevant, substrate concentrations. Insulin and spermine both inhibit the phosphorylation of the enzyme, possibly by increasing dephosphorylation. Tyrosine aminotransferase is phosphorylated in liver cell incubations but the rate is slow and insensitive to additions to the medium. No parallel changes in flux could be detected. Both enzymes are subject to complex regulatory mechanisms, short- and long-term. Their activities may be coordinated in vivo by control exerted at the level of the plasma membrane where both amino acids share the same transport processes. Determination of the control coefficients for the several components indicates that membrane transport may be a major limitation on flux.

Animals↗

Protamine pretreatment attenuation of hemodynamic and hematologic effects of heparin-protamine interaction. A prospective randomized study in human beings undergoing aortic reconstructive surgery.

Hemodynamic and hematologic responses to protamine sulfate reversal of heparin's anticoagulant effects were studied in 15 consecutive randomized patients undergoing aortic reconstructive surgery. In a double-blinded manner, patients were pretreated with either normal saline solution (n = 8) or protamine (0.75 mg/kg/3 min, n = 7) 5 minutes before heparinization (150 IU/kg). After aortic grafts were placed, protamine (1.5 mg/kg/3 min) was administered intravenously to reverse the heparin. Arterial blood pressure, heart rate, pulmonary artery and capillary wedge pressure, central venous pressure, and cardiac output were monitored, as were platelet count, white blood cell count, activated clotting time, total hemolytic complement levels, and C3a levels. Calculated parameters included systemic vascular resistance and pulmonary vascular resistance. Pretreatment with protamine compared with saline solution prevented the hypotension (+6 vs. -16 mm Hg, p less than 0.05) and declining pulmonary artery pressure (+1 vs. -7 mm Hg, p less than 0.01) observed with protamine reversal of heparin. Significant differences between the two groups in central venous pressure and pulmonary vascular resistance were of less clinical relevance. Protamine pretreatment lessened the thrombocytopenia found during reversal compared with saline-pretreated patients although the difference was not statistically significant. Minimal hypotension occurring after protamine pretreatment alone was not accompanied by hemodynamic or hematologic changes, other than decreased heart rate.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

Effects of continuous intravenous infusions of nitroglycerin on canine systemic and hind limb microcirculatory hemodynamics.

Systemic and hind limb hemodynamics were assessed in anesthetized dogs during continuous 30-minute intravenous infusions of nitroglycerin at 1, 5, 10, and 25 micrograms/kg/min. Nitroglycerin at 1 microgram/kg/min redistributed hind limb blood flow; hind limb arteriovenous shunting increased from 5.6% +/- 4.0% to 17.8% +/- 7.4% after 30 minutes (p less than 0.01); absolute hind limb shunt flow increased from 12 +/- 10 ml/min to 31 +/- 26 ml/min at 10 minutes (p less than 0.01); whereas hind limb nutrient blood flow decreased from 184 +/- 81 ml/min to 150 +/- 55 ml/min and 132 +/- 32 ml/min, respectively, at 10 and 30 minutes (p less than 0.05). Such hind limb blood flow redistribution was absent during infusion of all other nitroglycerin dosages. Total catecholamines increased at 30 minutes during both 1 and 10 micrograms/kg/min nitroglycerin infusions (p less than 0.05) with perhaps a slightly greater catecholamine response to 10 micrograms/kg/min after 30 minutes (0.05 less than p less than 0.10). The renin-angiotensin response at 3 minutes differed between nitroglycerin infusions of 1 and 10 micrograms/kg/min with an initial significant reduction from baseline in plasma renin activity at the lower dose compared with a significant increase from baseline in plasma activity at the higher dose. Nitroglycerin did not increase femoral artery flow or cardiac output and did not lower total peripheral vascular resistance at any dose studied. Despite this, arterial pressure and cardiac work were reduced at all nitroglycerin doses tested. Massive volume loading prevented the anticipated blood pressure reduction and blunted the expected cardiac work reduction during nitroglycerin infusions of 10 micrograms/kg/min. This study demonstrates that nitroglycerin is not a potent peripheral arteriolar vasodilator, 1 microgram/kg/min nitroglycerin infusions increase hind limb arteriovenous shunting and decrease hind limb nutrient blood flow, and myocardial work and arterial pressure reductions during nitroglycerin infusions appear to be caused by mechanisms other than generalized peripheral arterial dilation.

Animals↗

Absence of effects of dietary wheat bran on the activities of some key enzymes of carbohydrate and lipid metabolism in mouse liver and adipose tissue.

1. The effects of a 100 g/kg dietary substitution of wheat bran on the body-weight gain, food consumption and faecal dry weight of mice given a high-sucrose diet and on the activities of some key enzymes of carbohydrate and lipid metabolism in liver and adipose tissue were studied. 2. Wheat bran had no effect on body-weight gain, food consumption or faecal dry weight. 3. Wheat bran had no effect on the activities of hepatic glucose-6-phosphate dehydrogenase (EC 1.1.1.49), 6-phosphogluconate dehydrogenase (EC 1.1.1.44), malate dehydrogenase (oxaloacetate-decarboxylating) (NADP+) (EC 1.1.1.40), ATP-citrate (pro-3S)-lyase (EC 4.1.3.8), pyruvate kinase (EC 2.7.1.40) and fructose-1,6-bisphosphatase (EC 3.1.3.11). The activity of hepatic 6-phosphofructokinase (EC 2.7.1.11) increased but only when expressed on a body-weight basis. 4. Wheat bran had no effect on the activities of adipose tissue glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, malate dehydrogenase (oxaloacetate-decarboxylating) (NADP+), ATP-citrate (pro-3S)-lyase, hexokinase (EC 2.7.1.1), 6-phosphofructokinase and pyruvate kinase. 5. These results suggest that unlike guar gum and bagasse, wheat bran does not change the flux through some pathways of lipogenesis in liver and adipose tissue when mice are given high-sucrose diets.

Adipose Tissue↗

Prostacyclin treatment of ischemic ulcers and rest pain in unreconstructible peripheral arterial occlusive disease.

The efficacy of prostacyclin (PGI2) treatment was assessed in 26 patients with surgically unreconstructible atherosclerotic arterial occlusive disease of the lower extremity manifested by nonhealing ulcers and rest pain. Patients were randomized to receive a 72-hour intravenous infusion of PGI2 (6 ng/kg/min, n = 13) or placebo (n = 13). Ulcer size was measured by photographic planimetry, and rest pain was graded by blinded objective scoring at monthly intervals for 6 months. Ulcer size increased 64% in PGI2-treated patients and 22% in placebo-treated patients by 1 month after infusion. Rest pain decreased slightly in both PGI2 and placebo groups. At the conclusion of the study, 54% of placebo-treated patients and 31% of PGI2-treated patients had a positive treatment response, indicated by at least a 20% decrease in ulcer size and a 33% decrease in rest pain. PGI2 infusion did not improve the high-placebo response rate seen in these patients with severely ischemic extremities. These results emphasize the importance of placebo-controlled studies, even in patients with unreconstructible arterial disease.

Aged↗

Endothelial cell seeding efficiency onto expanded polytetrafluorethylene grafts with different coatings.

Initial adherence of 111Indium-oxine labeled, cultured canine venous endothelial cells to expanded polytetrafluorethylene (ePTFE) grafts was evaluated using different precoatings of the surface or different pretreated cells. The precoatings evaluated consisted of blood for 5 (Group I) or 15 min (Group II), fibronectin (Group IV), and cryoprecipitate (Group VI). In addition to endothelial cells subcultured immediately before labeling, cells subcultured 48 hours prior to labeling and kept in suspension were studied. Such cells were evaluated on grafts precoated with blood for 5 min (Group III) and fibronectin precoated surfaces (Group V). The amount of fibronectin that adhered to the graft surface with our technique was less den 1%. Seeding efficiency was higher with the blood precoated surfaces (5.3, 3.0, 2.2% in Groups I, II and III respectively) than fibronectin (1.7%, 1.7% in Groups IV, V respectively) or cryoprecipitate (1.9%, Group VI) precoated surfaces. No significant difference between cells immediately subcultured and these kept in suspension for 48 hours regarding adherence to blood (5.3 vs. 2.2%) or fibronectin precoated (1.7 vs. 1.7%) grafts was documented. The seeding efficiency of ePTFE grafts is low and further efforts to improve adherence must be made. From the present studies we recommend the use of blood precoating for about 5 min followed by cell incubation for approximately 10 min in order to achieve optimal seeding efficiency.

Animals↗

Attenuation of hemodynamic and hematologic effects of heparin-protamine sulfate interaction after aortic reconstruction in a canine model.

This investigation documented the effect of protamine sulfate pretreatment on the adverse hemodynamic and hematologic sequela of rapid intravenous protamine administration in heparinized dogs having undergone prior implantation of aortic prostheses. Fourteen dogs underwent infrarenal aortic replacement with 6 mm inner diameter by 7 cm knitted Dacron double-velour grafts. Carotid arterial, central venous, and pulmonary arterial catheters were placed for continuous hemodynamic monitoring. All dogs were adequately anticoagulated with heparin (150 IU/kg) and reversed with protamine (1.5 mg/kg/10 sec) after graft insertion. Pretreatment regimens studied included normal saline (n4), protamine 0.75 mg/kg (n5), and protamine 2.25 mg/kg in three divided doses of 0.75 mg/kg each (n5). All pretreatment agents were administered 3 minutes before heparinization. Blood pressure (BP), heart rate (HR), pulmonary artery pressure (PAP) and cardiac output (CO) were measured. Hematologic assessments included platelet count (PC), leukocyte count, thrombin clotting time, and total hemolytic complement. Significant salutory effects were associated with protamine pretreatment regarding BP, HR, PAP, CO, and PC. It is concluded that adverse hemodynamic effects of protamine reversal are blocked and certain hematologic effects are reduced with protamine pretreatment before heparinization and its reversal in this canine experimental model.

Animals↗

The effect of pyridoxine deficiency on the metabolism of the aromatic amino acids by isolated rat liver cells.

The total activity of three key enzymes and the flux through eight steps of aromatic amino acid metabolism have been determined in liver cells isolated from rats fed either control or pyridoxine-free diet for 5-6 weeks. The pyridoxine-free diet caused a decrease in the catabolism of tyrosine and phenylalanine because of a drop in the flux through tyrosine aminotransferase. This decrease of expressed cellular tyrosine aminotransferase activity can be fully explained in terms of loss of cofactor. Larger decreases in the catabolism of tryptophan were seen after pyridoxine deprivation. The decreased extent of tryptophan catabolism can be solely attributed to loss of cofactor or increased degradation of kynureninase. Inhibition of tryptophan 2,3-dioxygenase was seen in pyridoxine deficiency, probably because of the buildup of the kynurenine metabolites. The control strength of kynureninase, for flux through kynureninase, was calculated to be less than or equal to 0.004, but 0.41 after pyridoxine deprivation. The sensitivity of the three pathways to pyridoxine deprivation is interpreted and discussed in terms of the different affinities for pyridoxal phosphate and the control strengths of the pyridoxal phosphate-dependent enzymes, tyrosine aminotransferase and kynureninase.

Amino Acids↗

The metabolism of L-phenylalanine and L-tyrosine by liver cells isolated from adrenalectomized rats and from streptozotocin-diabetic rats.

Flux through, and maximal activities of, key enzymes of phenylalanine and tyrosine degradation were measured in liver cells prepared from adrenalectomized rats and from streptozotocin-diabetic rats. Adrenalectomy decreased the phenylalanine hydroxylase flux/activity ratio; this was restored by steroid treatment in vivo. Changes in the phosphorylation state of the hydroxylase may mediate these effects; there was no significant change in the maximal activity of the hydroxylase. Tyrosine metabolism was enhanced by adrenalectomy; this was not related to any change in maximal activity of the aminotransferase. Steroid treatment increased the maximal activity of the aminotransferase. Both acute (3 days) and chronic (10 days) diabetes were associated with increased metabolism of phenylalanine; insulin treatment in vivo did not reverse these changes. Although elevated hydroxylase protein concentration was a major factor, changes in the enzyme phosphorylation state may contribute to differences in phenylalanine degradation in the acute and chronic diabetic states. Tyrosine metabolism, increased by diabetes, was partially restored to normal by insulin treatment in vivo. These changes can, to a large extent, be interpreted in terms of changes in the maximal activity of the aminotransferase.

Acute Disease↗

Brachiocephalic arterial occlusions and stenoses. Manifestations and management of complex lesions.

Complex stenotic and occlusive lesions involving multiple brachiocephalic arteries were encountered in 17 symptomatic patients, 25 to 76 years of age. Symptoms included hemispheric transient ischemic attacks (16), visual symptoms (ten), global cerebral ischemia (11), true syncope (six), upper extremity ischemic symptoms (eight), and frank tissue loss (one). Of 68 brachiocephalic arteries, 53 exhibited hemodynamically significant stenoses, including 21 that were totally occluded. Transthoracic surgical reconstruction consisted of bypass grafting (11), innominate artery endarterectomy (five), or proximal left common carotid endarterectomy with reimplantation into the contralateral carotid artery (one). There were no operative deaths and only one transient perioperative neurologic deficit. All patients had relief of symptoms. When multiple brachiocephalic arterial occlusions and stenoses preclude standard cervical reconstructive procedures, direct transthoracic reconstruction is appropriate and may be undertaken with acceptable risk in properly selected patients.

Adult↗