Field efficacy of sweet corn hybrids expressing a Bacillus thuringiensis toxin for management of Ostrinia nubilalis (Lepidoptera: Crambidae) and Helicoverpa zea (Lepidoptera: Noctuidae).

Field studies were done in 1995-1996 to assess the efficacy of three sweet corn hybrids that express the Bacillus thuringiensis (Bt) toxin, CrylAb, against two lepidopteran pests, Ostrinia nubilalis (Hubner) and Helicoverpa zea (Boddie). The Bt hybrids tested were developed by Novartis Seeds, using the event BT-11, which expresses Bt toxin in green tissue as well as reproductive tissues including the tassel, silk, and kernel. Bt hybrids were compared with a standard non-Bt control or the non-Bt isoline for each hybrid; none of the hybrids were treated with insecticides during the study. Hybrid efficacy was based on larval control of each pest, as well as plant or ear damage associated with each pest. In both years, control of O. nubilalis larvae in primary ears of all Bt hybrids was 99-100% compared with the appropriate non-Bt check. Plant damage was also significantly reduced in all Bt hybrids. In 1996, control of H. zea in Bt hybrids ranged from 85 to 88% when compared with the appropriate non-Bt control. In 1996, a University of Minnesota experimental non-Bt hybrid (MN2 x MN3) performed as well as the Bt hybrids for control of O. nubilalis. Also, in 1996, two additional University of Minnesota experimental non-Bt hybrids (A684su X MN94 and MN2 X MN3) performed as well as Bt hybrids for percent marketable ears (ears with no damage or larvae). In addition, compared with the non-Bt hybrids, percent marketable ears were significantly higher for all Bt hybrids and in most cases ranged from 98 to 100%. By comparison, percent marketable ears for the non-Bt hybrids averaged 45.5 and 37.4% in 1995 and 1996, respectively. Results from the 2-yr study strongly suggest that Bt sweet corn hybrids will provide high levels of larval control for growers in both fresh and processing markets. Specifically, Bt sweet corn hybrids, in the absence of conventional insecticide use, provided excellent control of O. nubilalis, and very good control of H. zea. However, depending on location of specific production regions, and the associated insect pests of sweet corn in each area, some insecticide applications may still be necessary.

A compliant tubular device to study the influences of wall strain and fluid shear stress on cells of the vascular wall.

PURPOSE: Cellular constituents of the blood vessel wall are continuously subjected, in vivo, to both mechanical and hemodynamic forces, which elicit structural and biologic responses. We have developed a compliant tubular system, the vascular simulating device (VSD), that reproduces these forces, while supporting the attachment and the experimental manipulation of endothelial and smooth muscle cells. METHODS: The VSD consists of a compliant silicone rubber tube coupled to a pump system, which permits the simultaneous application of known levels of pressure and flow, to vascular wall cells cultured on the inner surface of the tube. Seeded cells can be monitored visually under phase contrast or fluorescent optics, as well as harvested and analyzed for biologic responses. RESULTS: The elastic modulus and compliance of the silicone rubber tube are similar to those of canine and human arteries. Endothelial and smooth muscle cells cultured on the lumenal surface of the tubes remain attached and viable after subjecting them to physiologic pulsatile flow and cyclic strain. CONCLUSION: The VSD makes it possible to approximate, in vitro, those forces encountered by vascular wall cells, in vivo and therefore may make it possible to determine whether specific combinations of mechanical and hemodynamic forces are causally associated with specific vascular diseases.

Biaxial elastic properties of rat arteries in vivo: influence of vascular wall cells on anisotropy.

There is no consensus as to the degree of arterial anisotropy or to its relationship to vascular cell function. Given the relevance of the isotropic assumption in formulating elasticity models, reliable measures of biaxial displacements are needed. In this study, a video motion analyzer (VMA) was used to describe the biaxial in vivo dynamic elasticity of 22 carotid arteries and 5 abdominal aortas in 27 rats. The influence of vascular cell function was also examined by subjecting six rats to a photosensitive drug, chloroaluminum sulfonated phthalocyanine (CASPc), which is focally cytotoxic on activation by laser. Circumferential compliance (Ccirc) was greater than longitudinal compliance (Clong) for all vessels. Compliance pressure curves were nonlinear, and biaxial displacements were in phase. The circumferential elastic modulus was less than the longitudinal modulus at common stresses. CASPc + laser reduced Ccirc but not Clong, thus altering Poisson's ratio. In conclusion, rat arteries are biaxially, nonlinearly elastic and anisotropic in vivo. Vascular cells modulate Poisson's ratio by influencing Ccirc.

Longterm study of a compliant biological vascular graft.

Clinically implanted prostheses of biological origin have recently been shown to develop aneurysms within several years. To study this process, recently developed bovine heterografts that were implanted in canine ilio-femoral arteries for 27 to 45 months, were studied in vivo and in vitro. Seven out of seven grafts were patent with only one showing evidence of focal aneurysm. Measurements of mechanical properties, including water permeability, compliance, and burst pressure, and of heat shrink temperature and dry weight were obtained before and after a period of controlled exposure to bacterial collagenase; all data suggested that much of the original graft had been replaced with host tissue. However, enzyme susceptibility was less than that of fresh bovine arteries, indicating that at least some of the grafts' crosslinked collagen was preserved. The compliance of these explants was similar to that reported for autogenous vein. Histological examination of the graft wall revealed cellular "intimal" and adventitial zones containing host-generated collagen surrounding a central zone devoid of cellular infiltration, which appears to be unmodified graft "media" highly resistant to degradation. We conclude that, when properly processed, biological prostheses can act as a bioresorbable scaffold for the orderly replacement of structural elements during healing, promoting continued mechanical integrity of the graft. These promising results encourage the further development of prostheses of biological origin.

A laboratory model to quantitate the resistance of collagen vascular grafts to biodegradation.

Recent reports have shown that despite extensive preclinical testing, vascular grafts of biological origin undergo severe biodegradation and aneurysm formation after two or more years of implantation in man. The purpose of this study was to develop a laboratory model to quantitate and correlate the stability of crosslinked collagen grafts in vitro and in vivo. This resistance to biodegradation was assessed by measuring changes in suture pullout force and sample weight in response to controlled digestion with bacterial collagenase, in 0.5-cm-long cylindrical graft segments (chemically processed bovine carotid artery and human umbilical cord vein) that were implanted in the rat subcutis for 2 to 12 weeks. Scar tissue was removed from the explants by brief enzymatic digestion, a process that was inhibited when graft segments had become infected. Changes in dry weight were more consistent than were changes in wet weight; drying the graft segments had no effect on their degradation in vivo or in vitro. Intact cylindrical rings suffered somewhat less damage than did opened, flattened cylinders. Graft degradation increased markedly with implantation time, and was detected after only 3 weeks. We conclude that the rat subcutis model, when combined with controlled enzymatic digestion, first to remove scar tissue and then to challenge structural integrity, provides an accelerated assay by which to predict the stability of collagen vascular grafts.

Prediction of aneurysm formation in vascular grafts of biologic origin.

Other than review of clinical experience, no assay exists that can reliably predict the long-term potential for aneurysm formation in an arterial prosthesis of biologic origin. Since mural degeneration probably results from proteolytic digestion, an in vitro assay was devised that used graft perfusion with 1% collagenase to induce rapid changes in mechanical properties. The effect of enzyme on graft diameter, compliance, permeability, and burst pressure was measured in ficin-digested, adipoyl chloride and glutaraldehyde-tanned bovine carotid artery and glutaraldehyde-tanned human umbilical vein. Both grafts have recently been reported to have a significant incidence of aneurysm within several years of implantation. Compliance and diameter were also measured noninvasively in patients with bovine carotid artery and human umbilical vein for more than 40 weeks after implantation. In vitro, the response to enzyme could be categorized into three groups. In group III, a diameter increase of more than 14% was associated with a significantly decreased compliance, and this paralleled the results found in aneurysmal grafts in vivo. In both grafts there was a strong correlation between postenzyme compliance change and initial compliance, loss of compliance being significantly greater in grafts with group III responses (p less than 0.01). This response may be a good predictor of a graft's overall susceptibility to aneurysmal degeneration, and initial compliance measurement may effectively identify inadequate fixation. Thus measurement of compliance may prove useful in quality control of a fixation process used in mass production. In conclusion, measurement of mechanical properties of biologic vascular grafts before and after collagenase exposure forms the basis for an effective in vitro assay of aneurysm susceptibility.

The evolution of morphologic and biomechanical changes in reversed and in-situ vein grafts.

A comparative study of experimental reversed (RV) and in-situ (INS) vein grafts with respect to the evolution of morphologic and compliance characteristics was done in a canine model. In addition, the compliance characteristics in a series of human INS vein grafts were recorded as a function of time after operation. At 6 months after implantation, all experimental grafts displayed well-developed intimal hyperplasia. There was no significant difference in either absolute intimal thickness (INS 0.133 +/- 0.09 mm vs. RV 0.085 +/- 0.06 mm; NS) nor in the percentage of the total wall thickness occupied by the intima when experimental INS grafts were compared with RV grafts after 6 months. Similarly, compliance values of INS and RV vein grafts were similar at all time intervals examined up to 6 months after operation. Thirty-three human INS vein grafts had a mean compliance value of 1.74 +/- 0.72 (percent radial changes per mmHg X 10(-2) at a median postoperative interval of 14 weeks. This value did not differ significantly from those measured in the INS vein grafts. Although all vein grafts examined retained their native viscoelastic properties, this study suggests that functioning human INS vein grafts are less compliant than previously suspected on the basis of prior ex-vivo and clinical studies of RV saphenous vein grafts. The purported clinical superiority of the INS vein graft cannot be explained on the basis of superior biomechanical performance or failure to develop intimal hyperplasia.

Effect of compliance mismatch on vascular graft patency.

The hypothesis that a mismatch in compliance between a vascular graft and its host artery is detrimental to graft patency was tested by implanting paired arterial autografts, prepared with differential glutaraldehyde fixation of carotid arteries in the femoral arteries of dogs. These grafts differed only in circumferential compliance: they were 100% (compliant) vs. 40% (stiff) as compliant as the host artery. Their flow surfaces were equivalent, as determined by physicochemical measurements and scanning electron microscopy; both lacked viable cells, as determined by in vitro cell culture. In 14 dogs, eight stiff and two compliant grafts became occluded within 3 months, the latter doing so within 24 hours after their contralateral counterparts. Cumulative patencies were 85% and 37% for compliant and stiff grafts, respectively (p less than 0.05) and 100% and 43%, excluding the two dogs with bilateral graft failures (p less than 0.01). We conclude that even with near optimal flow surfaces, compliance mismatch is deleterious to graft patency.

Noninvasive measurements of nonlinear arterial elasticity.

To describe fully the compliance characteristics of an artery with nonlinear elastic properties, measurements must be obtained over a wide range of pressures. Furthermore, repeated measurements, as required in temporal studies of arterial implants, require that the measuring technique be noninvasive. The application of a pulsed ultrasound echo-tracking device is described, which fulfills both criteria. Nonlinear compliance-pressure (CP) curves were obtained from the femoral arteries of dogs, with the use of halothane anesthesia to vary systemic pressure, and were used to compare the gross elastic properties of different vessels. Studies using controlled hemorrhage or the vasoactive drugs, nitroprusside and levarterenol (norepinephrine), were used to verify that the CP curves obtained during halothane anesthesia did not reflect varying degrees of smooth muscle activation. However, surgical exposure did temporarily reduce arterial compliance at pressures between 60 and 140 mmHg. The effect of vasoactive intervention and of postsurgical changes in arterial or graft compliance can thus be quantitated by use of CP curves or by comparing incremental elastic moduli, which can also be estimated from the noninvasively derived measurements.

Modification of the Haemonetics Cell Saver for optional high flow rate autotransfusion.

Intraoperative autotransfusion is a technique well-suited to major vascular surgery. It is most effective when salvage and reinfusion of shed blood can be accomplished at flow rates compatible with the degree of hemorrhage encountered in both elective and emergency procedures. Appropriate equipment modifications can render commercially available autotransfusion devices safer and more effective in the management of intraoperative blood loss. The Cell Saver, a device which concentrates and washes salvaged red blood cells, is limited in its potential as an autotransfusion device because of its slow reinfusion rate. A modification was devised which expands the flow capabilities of the Cell Saver and allows rapid reinfusion of autologous whole blood. The modified blood circuit has been employed in 10 major vascular cases with favorable results, thus demonstrating its efficacy in the management of massive hemorrhage during vascular repair. Guidelines for the safe and effective use of the modified unit are stressed.

Intraoperative autotransfusion in major vascular surgery.

The use of intraoperative autotransfusion provides a safe and cost-effective means of salvaging operative blood loss and reducing or eliminating the use of stored homologous bank blood with its inherent difficulties and risks. The risk of disease transmission or various reactions is minimized. Autotransfusion provides a readily available, more physiologic, and at times life-saving source of blood for patients with rare blood types or patients in whom time does not permit adequate cross-matching. This technique is acceptable to most sects of Jehovah's Witnesses, who normally refuse homologous blood. Our experience during the past six years with autotransfusion in major vascular surgery reveals a mean slavage equivalent to five units of blood loss, and avoidance of using any bank blood in almost half of elective patients. No significant problems occurred due to hemolysis, coagulation abnormalities, or particulate/air emboli, nor any morbidity or mortality specifically related to autotransfusion. We conclude that wider and more frequent use of autotransfusion technics is appropriate.