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Biomedical subjects

R Nowygrod

Publications and source records attributed to R Nowygrod.

At least 37 records · Page 2Linked to original sources

Canine choledochotomy closure with diode laser-activated fibrinogen solder.

BACKGROUND: An alternative to mechanical stapling or hand suturing is needed to permit laparoscopic common bile duct exploration. We evaluated the strength and healing characteristics of canine choledochotomies sealed with a fibrinogen solder and a diode laser. METHODS: After creation of a 0.5 cm longitudinal choledochotomy, the edges were coapted with forceps, and a fibrinogen solder mixed with indocyanine green dye was applied. The solder was sealed in place with an 810 nm diode laser (125 W/cm2). RESULTS: Immediate mean leakage pressure was 264 +/- 7 mm Hg compared with 83 +/- 66 mm Hg in suture controls. This increased to 364 +/- 115 mm Hg at 2 days and was more than 510 mm Hg at 7 days. On histologic examination rapid reabsorption of the solder with no signs of inflammation or stenosis was seen. No episodes of dehiscence or peritonitis occurred. CONCLUSIONS: Laser soldering provides a watertight choledochotomy closure with adequate immediate strength allowing a reliable, technically feasible common bile duct exploration via a laparoscopic approach.

Animals↗

Laser-assisted fibrinogen bonding of umbilical vein grafts.

Despite success with autologous tissue welding, laser welding of synthetic vascular prostheses has not been possible. The graft material appears inert and fails to allow the collagen breakdown and electrostatic bonding that results in tissue welding. To develop a laser welding system for graft material, we repaired glutaraldehyde-tanned human umbilical cord vein graft incisions using laser-assisted fibrinogen bonding (LAFB) technology. Modified umbilical vein graft was incised transversely (1.2 cm). Incisions were repaired using sutures, laser energy alone, or LAFB. For LAFB, indocyanine green dye was mixed with human fibrinogen and the compound applied with forceps onto the weld site prior to exposure to 808 nm diode laser energy (power density 4.8 W/cm 2). Bursting pressures for sutured repairs (126.6 +/- 23.4 mm Hg) were similar to LAFB anastomoses (111.6 +/- 55.0 mm Hg). No evidence of collateral thermal injury to the graft material was noted. In vivo evaluation of umbilical graft bonding with canine arteries demonstrates that LAFB can reliably reinforce sutured anastomoses. The described system for bonding graft material with laser exposed fibrinogen may allow creation or reinforcement of vascular anastomoses in procedures where use of autologous tissue is not feasible.

Anastomosis, Surgical↗

Regulation of human mononuclear phagocyte migration by cell surface-binding proteins for advanced glycation end products.

Nonenzymatic glycation of proteins occurs at an accelerated rate in diabetes and can lead to the formation of advanced glycation end products of proteins (AGEs), which bind to mononuclear phagocytes (MPs) and induce chemotaxis. We have isolated two cell surface-associated binding proteins that mediate the interaction of AGEs with bovine endothelial cells. One of these proteins is a new member of the immunoglobulin superfamily of receptors (termed receptor for AGEs or RAGE); and the second is a lactoferrin-like polypeptide (LF-L). Using monospecific antibodies to these two AGE-binding proteins, we detected immunoreactive material on Western blots of detergent extracts from human MPs. Radioligand-binding studies demonstrated that antibody to the binding proteins blocked 125I-AGE-albumin binding and endocytosis by MPs. Chemotaxis of human MPs induced by soluble AGE-albumin was prevented in a dose-dependent manner by intact antibodies raised to the AGE-binding proteins, F(ab')2 fragments of these antibodies and by soluble RAGE. When MP migration in response to N-formyl-Met-Leu-Phe was studied in a chemotaxis chamber with AGE-albumin adsorbed to the upper surface of the chamber membrane, movement of MPs to the lower compartment was decreased because of interaction of the glycated proteins with RAGE and LF-L on the cell surface. The capacity of AGEs to attract and retain MPs was shown by implanting polytetrafluoroethylene (PTFE) mesh impregnated with AGE-albumin into rats: within 4 d a florid mononuclear cell infiltrate was evident in contrast to the lack of a significant cellular response to PTFE with adsorbed native albumin. These data indicate that RAGE and LF-L have a central role in the interaction of AGEs with human mononuclear cells and that AGEs can serve as a nidus to attract MPs in vivo.

Animals↗

Restoration of the cAMP second messenger pathway enhances cardiac preservation for transplantation in a heterotopic rat model.

Current organ preservation strategies subject graft vasculature to severe hypoxia (PO2 approximately 20 Torr), potentially compromising vascular function and limiting successful transplantation. Previous work has shown that cAMP modulates endothelial cell (EC) antithrombogenicity, barrier function, and leukocyte/EC interactions, and that hypoxia suppresses EC cAMP levels. To explore the possible benefits of cAMP analogs/agonists in organ preservation, we used a rat heterotopic cardiac transplant model; dibutyryl cAMP added to preservation solutions was associated with a time- and dose-dependent increase in the duration of cold storage associated with successful graft function. Preservation was also enhanced by 8-bromo-cAMP, the Sp isomer of adenosine 3',5'monophosphorothioate, and types III (indolidan) and IV (rolipram) phosphodiesterase inhibitors. Neither butyrate alone nor 8-bromoadenosine were effective, and the cAMP-dependent protein kinase antagonist Rp isomer of adenosine 3',5'monophosphorothioate prevented preservation enhancement induced by 8-bromo-cAMP. Grafts stored with dibutyryl cAMP demonstrated a 5.5-fold increase in blood flow and a 3.2-fold decreased neutrophil infiltration after transplantation. To explore the role of cAMP in another cell type critical for vascular homeostasis, vascular smooth muscle cells were subjected to hypoxia, causing a time-dependent decline in cAMP levels. Although adenylate cyclase activity was unchanged, diminished oxygen tensions were associated with enhanced phosphodiesterase activity (59 and 30% increase in soluble types III and IV activity, respectively). These data suggest that hypoxia or graft ischemia disrupt vascular homeostasis, at least in part, by perturbing the cAMP second messenger pathway. Supplementation of this pathway provides a new approach for enhancing cardiac preservation, promoting myocardial function, and maintaining vascular homeostatic properties.

3',5'-Cyclic-AMP Phosphodiesterases↗

Survey of the distribution of a newly characterized receptor for advanced glycation end products in tissues.

Advanced glycation end products (AGEs), the final products of nonenzymatic glycation and oxidation of proteins, are found in the plasma and accumulate in the tissues during aging and at an accelerated rate in diabetes. A novel integral membrane protein, termed receptor for AGE (RAGE), forms a central part of the cell surface binding site for AGEs. Using monospecific, polyclonal antibody raised to human recombinant and bovine RAGE, immunostaining of bovine tissues showed RAGE in the vasculature, endothelium, and smooth muscle cells and in mononuclear cells in the tissues. Consistent with these data, RAGE antigen and mRNA were identified in cultured bovine endothelium, vascular smooth muscle, and monocyte-derived macrophages. RAGE antigen was also visualized in bovine cardiac myocytes as well as in cultures of neonatal rat cardiac myocytes and in neural tissue where motor neurons, peripheral nerves, and a population of cortical neurons were positive. In situ hybridization confirmed the presence of RAGE mRNA in the tissues, and studies with rat PC12 pheochromocytes indicated that they provide a neuronal-related cell culture model for examining RAGE expression. Pathological studies of human atherosclerotic plaques showed infiltration of RAGE-expressing cells in the expanded intima. These results indicate that RAGE is present in multiple tissues and suggest the potential relevance of AGE-RAGE interactions for modulating properties of the vasculature as well as neural and cardiac function, prominent areas of involvement in diabetes and in the normal aging process.

Animals↗

Laser bonding of secondary bronchi with solvent--detergent-treated cryoprecipitate.

Management of bronchopleural fistula is a challenging clinical problem. Laser-assisted cryoprecipitate bonding techniques offer a means to fix precisely tissue glues into the fistulae through a bronchoscopic approach. Analogous studies exist using fibrin glue with thrombin. Using a canine model, secondary bronchi were sealed with cryoprecipitate made from solvent/detergent-treated plasma (treated to inactivate membrane-enveloped virus) mixed with indocyanine green (absorption 805 nm). Diode laser energy (emission 808 nm, 7.3 W/cm 2) was applied to the solder until dessication was observed. Leakage pressures (n = 7) ranged between 18 and 86 mmHg with a mean of 46 +/- 24 mmHg. Laser-assisted solder techniques provide a reliably strong seal over leaking bronchial stumps and use of dye enhancement prevents undesired collateral thermal injury to surrounding bronchial tissue. Solvent/detergent plasma, prepared by methods shown to inactivate large quantities of HIV, HBV, and HCV, is an effective source of cryoprecipitate and should allow widespread use of pooled human material in a clinical setting.

Animals↗

Comparison of laser-assisted fibrinogen-bonded and sutured canine arteriovenous anastomoses.

The effect of laser-assisted fibrinogen bonding (LAFB) on the development of intimal hyperplasia was studied with stress-strain profiles and histologic evaluation of canine arteriovenous fistulas (AVFs). In 19 animals femoral AVFs were created with an 808 nm diode laser after topical application of fibrinogen mixed with indocyanine green dye; in the contralateral limb a sutured AVF was created. The animals were divided into three groups. Group 1 dogs (n = 6) were killed serially up to 4 weeks after surgery to examine the healing of the anastomoses created with LAFB. Group 2 dogs (n = 6) were killed 1 month after surgery, and the fresh specimens were strained axially to produce a stress-strain profile graph. Group 3 dogs (n = 7) were killed 7 months after surgery, and the AVFs were infused with formalin under pressure and histologically prepared to allow comparison of the ratio of maximum to minimum intimal hypertrophy. Fibrinogen used for LAFB was resorbed during the first month after operation without evidence of foreign body reaction or inflammation. Tensile break force was not significantly different in the laser-bonded group (4.6 +/- 2.4 pounds) and the sutured group (4.3 +/- 1.7 pounds). The modulus (tensile break force per square inch), a measure of elasticity, identified the laser-bonded AVF (149 +/- 44 pounds per square inch) to be less rigid than the sutured AVF (203 +/- 35 pounds per square inch) (p less than 0.05). No significant differences in the degree of intimal hyperplasia were noted in any area of the anastomoses. Use of LAFB neither accelerates nor prevents intimal hyperplasia in a canine AVF model.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Hepatobiliary complications of polyarteritis nodosa.

Although polyarteritis nodosa (PAN) may result in thrombosis or aneurysm formation in any organ in the body, hepatobiliary complications are unusual. We reviewed seven cases that demonstrated the diagnostic difficulties and therapeutic options available in the management of hepatobiliary PAN. No consistent sign that indicated the severity of hepatobiliary PAN could be identified. In cases of thrombotic PAN, acalculus cholecystitis usually could be diagnosed preoperatively. Early tissue diagnosis and aggressive intervention are required for appropriate patient treatment. If the diagnosis is unclear, a preoperative muscle or skin biopsy specimen is often helpful in establishing a tissue diagnosis of PAN, even if no obvious pathologic condition is evident. Patients who undergo celiotomy for acalculus cholecystitis or peritoneal signs of an unclear origin should have tissue specimens (gallbladder wall, liver, or omentum) submitted for pathologic study. Angiography may be diagnostic preoperatively or when results of biopsies are equivocal. In addition, early angiography can define the extent of visceral involvement and permit control by embolization of hemorrhage secondary to aneurysm rupture. Awareness of the possibilities of thrombotic, ischemic, or bleeding complications from PAN allows more aggressive and rapid management of abdominal complaints, especially in patients who are receiving immunosuppressant therapy.

Adult↗

The accuracy of CT scanning in the diagnosis of abdominal and thoracoabdominal aortic aneurysms.

As CT scanning has evolved as a reliable clinical tool, the use of angiography in the diagnosis of aortic aneurysmal disease has diminished. Fewer than 25% of patients with aortic aneurysmal disease undergo aortic angiographic evaluation at our institution. A prospective clinical study was undertaken to assess the validity of this policy. One hundred patients with clinical or ultrasonographic evidence of aortic aneurysms were evaluated prospectively during the period July 1987 to December 1989. All patients underwent CT scanning as an initial evaluation. Patients were selected for angiography if they fulfilled any of the following criteria: radiographic evidence of thoracoabdominal or juxtarenal aneurysms, or horseshoe kidney; or clinical suggestion of renal artery stenosis, mesenteric arterial insufficiency, aortoiliac occlusive disease, or lower extremity aneurysmal disease. During this period 19 patients (19%) underwent both CT scanning and angiography. The indications for angiography were thoracoabdominal aneurysms (7), juxtarenal aneurysms (2), clinical evidence of mesenteric insufficiency (1) or renal insufficiency (2), evidence of lower extremity aneurysmal disease (3), or severe aortoiliac occlusive disease (4). Eighty-one patients (81%) underwent CT scanning as the only radiographic evaluation. No patient was adversely affected by elimination of angiographic evaluation. CT scanning revealed inflammatory aneurysms (4), retroaortic renal veins (2), and horseshoe kidney (1). This study suggests that most (81%) patients with aortic aneurysmal disease can be adequately evaluated by CT scanning, and that a very selective policy of angiographic evaluation is indicated.

Aged↗

A preliminary study of dye-enhanced laser photosclerosis.

Laser ablation of veins after injection of wavelength-specific dyes to enhance and localize energy absorption could provide a useful adjunct to current treatment options. To enhance the absorption of diode laser energy at 808 nm, ear veins of 41 rabbits were infused with 2 to 3 ml of indocyanine green dye (maximum absorption, 805 nm) and exposed for 2 to 20 seconds. Animals were killed between 0 and 28 days after operation. Discrete time intervals of laser exposure exist during which various-sized vessels can be ablated without significant thermal injury to the overlying tissue. Small vessels (0.2 mm in diameter) blanch after 2 to 3 seconds of exposure, whereas medium-sized vessels (2 mm in diameter) require 8 to 10 seconds. Vessels can be ablated with a power density as low as 11.1 W/cm2. Specimens taken immediately after laser exposure show vessel wall thinning and a reirradiation effect, created as laser energy initially absorbed by dye is reemitted. By the seventh day after operation, a brisk inflammatory response and acanthosis of the overlying epidermal layer develop. The lumen is partially filled by thrombus with cellular invasion. By postoperative day 28, the epidermal thickening and inflammatory reaction have resolved; the vessel walls are fibrotic. The use of low-power, air-cooled diode lasers, in conjunction with wavelength-specific dyes, may provide a simple, viable, and cosmetically appealing alternative to the treatment of superficial varicosities of the extremities.

Animals↗

Strength of laser vascular fusion: preliminary observations on the role of thrombus.

We hypothesized that autologous clot deposited on the luminal surface of laser vascular welds immediately after creation would produce higher time zero bursting pressures that could be achieved in welds perfused with saline alone. To test this hypothesis, we compared bursting pressures of welds created in isolated rabbit aortic segments 1) with saline perfusion only, 2) with blood perfusion, and 3) with blood perfusion followed by infusion of urokinase. Tissue welds with saline perfusion had a mean bursting pressure of 159 +/- 45 mm Hg; tissue welds following blood perfusion had a mean pressure of 262 +/- 29 mm Hg; tissue welds with blood perfusion followed by urokinase infusion had a mean bursting pressure of 187 +/- 35 mm Hg. The saline and urokinase groups were not significantly different. However, the blood-perfused group was significantly higher than both the saline group and the urokinase group. Thus, the addition of urokinase eliminates the beneficial effects noted after blood reperfusion. These observations suggest that the enhancement of weld strength following exposure to blood is due predominantly to the adherence of fibrin-platelet aggregates at the site of the weld.

Anastomosis, Surgical↗

Effects of a 2.15-micron laser on human atherosclerotic xenografts in vivo.

The thulium-holmium-chromium:yttrium-aluminum-garnet (THC:YAG) laser has a tissue effect similar to that of the CO2 laser, with the advantage of transmissibility through flexible fibers. The authors used a human-rabbit xenograft model to evaluate the thrombotic and healing responses of atherosclerotic vessels subjected to laser energy. Occluded atherosclerotic human coronary artery segments were recanalized in vitro by use of the THC:YAG laser. Destruction of plaque by the laser was achieved with minimal collateral thermal damage. These vascular segments were then transplanted into the rabbit abdominal aorta. The authors observed that the luminal surface of the lased vessels was more thrombogenic than that of the nonlased control vessels. However, occlusion of the lased vessels did not occur. Repair of laser-treated tissue progressed until a mature, nonthrombogenic fibrin-platelet aggregate was adherent to the luminal wall. Overall, the lased vessels behaved in a fashion similar to the nonlased control vessels. On the basis of these results, the authors believe that the THC:YAG laser may have use in human angioplasty.

Angioplasty, Laser↗

Tissue soldering by use of indocyanine green dye-enhanced fibrinogen with the near infrared diode laser.

Anastomoses welded by laser have been strengthened by applying a solder of fibrinogen combined with a laser energy absorbing dye (indocyanine green, maximum absorbance 805 nm) to the anastomotic site before continuous-wave diode laser exposure (808 +/- 1 nm, 4.8 W/cm2). Immediately after creation, the bursting pressures of welds created without fibrinogen (262 +/- 29 mm Hg, n = 11) were significantly less than repairs with fibrinogen (330 +/- 75 mm Hg, n = 11) (p less than 0.05). When repairs performed with fibrinogen were exposed to urokinase (25,000 IU) the bursting pressures were not significantly different from baseline (290 +/- 74 mm Hg, n = 5). Aortotomies closed by suture did not burst but leaked at pressures significantly below those of vessels closed by laser (165 +/- 9 mm Hg, n = 11) (p less than 0.01). Twenty-two repairs soldered with fibrinogen were incorporated into survival studies in rabbits and examined from 1 to 90 days after operation. No anastomotic ruptures, thromboses, or aneurysms were identified. Soldered sites rapidly regenerated a new intimal surface and healed by myofibroblast proliferation. No significant foreign body response was identified; the fibrinogen was resorbed. Laser soldering with exogenous fibrinogen is feasible without topical administration of additional clotting agents, significantly improves the bursting strength of primary laser welded anastomoses, and appears to result from urokinase-resistant fibrinogen cross-linking.

Anastomosis, Surgical↗

Dye-enhanced laser tissue welding.

For vascular anastomosis, use of topical photosensitizing dye enhances selective delivery of laser energy to target tissue, thus reducing the amount of collateral thermal injury and threshold power required for welding. For fluorescein isothiocyanate (FITC)--stained rabbit aorta in vitro, the threshold for tissue blanching was 15 seconds of 100 mW exposure of cw argon ion laser compared with 15 seconds at 300 mW for unstained tissue. The threshold power density needed for argon laser welding of abdominal aortotomies in rabbits in vivo was 3.8 W/cm2 with FITC and 7.6 W/cm2 without the dye. However, bursting pressures for the two groups (164 mm Hg with FITC, 147 mm Hg without FITC) were not significantly different. Histology revealed decreased collateral thermal damage in FITC-enhanced welds. Use of photosensitizing dyes for tissue welding is feasible and may allow arterial welding with lower power laser systems and cause less thermal trauma by lowering threshold power levels.

Animals↗

A fiberoptic compatible midinfrared laser with CO2 laser-like effect: application to atherosclerosis.

In theory, pulses of laser light in the 2-microns range should ablate tissue in a manner similar to that of the 10.6-microns CO2 laser with the added advantage of efficient transmission through flexible quartz fibers. Using 200-microseconds pulses of 2.15-microns thulium-holmium-chromium:YAG (THC:YAG) laser light, we were able to create 700-microns-diameter holes through calcific atherosclerosis in vitro. In vivo evaluation of thrombogenicity and healing was accomplished by exposing the luminal surface of rabbit aortas to the THC:YAG laser. Serial histologic examinations of laser-treated rabbit aortae revealed a time course of resolution of the lesions which was very similar to that observed with like-sized lesions created with the same amount of continuous wave CO2 energy. No significant differences in thrombogenicity nor healing response were noted. The excellent in vivo response observed is due in part to the pulsed nature of the THC:YAG laser output as well as to the efficient tissue absorption at the 2.15-microns wavelength. We feel that excellent ablative effects with minimal collateral thermal damage can be obtained through fiberoptic delivery systems by taking advantage of laser wavelengths corresponding to the infrared absorption peak of water in the 2-microns region and pulsed delivery of the laser energy.

Animals↗

Control of prosthetic bacterial infection: evaluation of an easily incorporated, tightly bound, silver antibiotic PTFE graft.

Despite the use of prophylactic antibiotics in vascular surgery, prosthetic infection rate remains 2-5%. Antibiotics bound to vascular prostheses can control experimentally induced infection but prolonged antibacterial activity has not been achieved. This study evaluates the in vivo efficacy and antibiotic retention of an easily prepared silver-antibiotic prosthesis. Prostheses were prepared by combining silver with oxacillin or amikacin using an organic solvent. After evaporation of the solvent, the graft was left impregnated with the antibiotic complex. In vivo retention studies were performed by implanting PTFE 110Ag-oxacillin prostheses in four canine abdominal aortas. When prostheses were explanted at 1 week, mean antibiotic retention was found to be 20% of original activity, higher than the mean inhibitory concentration for Staphylococcus aureus. In three groups of five dogs, 20 X 7-mm prostheses of PTFE alone, PTFE silver-oxacillin, or PTFE silver-amikacin were implanted in the abdominal aorta and the grafts were inoculated with 10(7) S. aureus of a known bacteriophage type, in a closed retroperitoneal pocket. The animals were sacrificed at 1 week and the prostheses were excised for quantitative bacterial culture. PFTE silver-oxacillin, and PTFE silver-amikacin prostheses had 1.7 X 10(2) and 2.0 X 10(2) colonies, respectively, significantly less (P less than 0.05) than controls (1.3 X 10(6) colonies). These data suggest that antibiotic prostheses can be easily prepared without binding agents. They retain the bound antibiotic for a prolonged period and are effective in reducing graft infection in a stringent direct contamination model.

Amikacin↗

Recent clinical experience with the vena cava filter.

In the past at our institution, insertion of the vena cava filter was recommended almost exclusively for recurrent pulmonary emboli in anticoagulated patients or in those with emboli and contraindications to anticoagulation. More recently, prophylaxis has become a frequent indication for patients with extensive deep vein thrombosis and contraindications to anticoagulation. Herein, we review a recent series of caval filter insertions to determine if increased utilization of this technique is justified in terms of morbidity and effectiveness. Twenty-one filters were inserted in 20 patients during a 1-year period. The operative mortality rate was 0 and satisfactory filter placement was achieved in 90 percent of patients. There were no documented instances of recurrent emboli or vena cava thrombosis. These data suggest that use of the filter is justified as prophylaxis for patients at high risk for an initial pulmonary embolus in a clinical setting where heparin therapy is not appropriate.

Adult↗