A comparison of melanin bleaching and azure blue counterstaining in the immunohistochemical diagnosis of malignant melanoma.

Distinguishing heavily pigmented melanocytes from melanophages on routine hematoxylin and eosin slides can be difficult. Melanin bleaching with potassium permanganate solution is a traditional means of removing melanin from tissues and can be used before immunohistochemical staining to remove any pigment that might be confused with the brown chromogen diaminobenzidine. Azure B stains melanin granules green-blue, easily contrasts with diaminobenzidine, and may be used as a counterstain on unbleached sections after immunohistochemical staining. To our knowledge, studies comparing melanin bleaching with azure B counterstaining in the immunohistochemical evaluation of malignant melanomas have not been performed. Paraffin sections from 33 heavily pigmented malignant melanomas were bleached with a 3.0-g/L potassium permanganate solution, immunohistochemically stained for S-100 and HMB-45, and counterstained with hematoxylin. Unbleached sections were similarly stained for S-100 and HMB-45 and counterstained with azure B. To establish optimal permanganate concentrations, a variable number of sections were bleached with lower permanganate concentrations ranging from 0.125 to 2.5 g/L. S-100 antigenicity was preserved at all permanganate concentrations, whereas HMB-45 antigenicity was abolished at concentrations of 0.5 g/L and greater. At permanganate concentrations from 0.125 to 0.5 g/L, both antigenicities were preserved; however, melanin was incompletely removed. Complications of bleaching included tissue damage and loss of cytologic detail. Positive immunohistochemical staining was observed in azure B counterstained sections. Azure B stained melanin greenblue and was easily distinguished from the brown diaminobenzidine chromogen, regardless of the antibody tested. Neither tissue damage nor loss of cytologic detail was observed. We conclude that the use of azure B counterstaining is superior to permanganate bleaching in the histologic evaluation of heavily pigmented cutaneous malignant melanomas.

Bradycardia induced by intravascular versus direct stimulation of the vagus nerve.

BACKGROUND: Electrical stimulation of the parasympathetic nervous system results in slowing of the heart. We sought to determine whether cardiac vagal efferent axons can be stimulated adequately to induce bradycardia without disturbing the integrity of the thorax. METHODS: Cardiodepressor effects elicited by direct stimulation of a vagus nerve in anesthetized dogs and pigs were compared with those generated when the same nerve was stimulated indirectly through bipolar electrodes placed in the adjacent superior vena cava. RESULTS: The heart rate of dogs decreased by about 80% when electrical stimuli were delivered to the right thoracic vagus at the level of the thoracic outlet through bipolar electrodes placed either in the adjacent superior vena cava (intravascular method) or directly on the nerve (direct method). Maximal responses were achieved with 10-V, 5-ms, and 20-Hz stimuli. In anesthetized pigs, similar bradycardia occurred when the right cervical vagus or the right cranial thoracic vagus was stimulated either directly or indirectly through the intravascular method. Atrial dysrhythmias occurred when the stimulating electrodes were placed by either method within 1 cm of the right atrium in both animal models. CONCLUSIONS: Controlled bradycardia can be induced during operation without the risk of generating cardiac dysrhythmias using electrical stimuli (10 V, 5 ms, and 10 to 20 Hz) delivered to the right cervical vagus nerve or the right cranial thoracic vagus nerve through adjacent intravascular electrodes.

Repair of soft tissue to bone using a biodegradable suture anchor.

The medial collateral ligaments of 18 New Zealand rabbits were surgically detached from bone. In one knee, the ligament was repaired using a biodegradable suture anchor composed of a co-polymer of lactic and glycolic acid. The contralateral medial collateral ligament was not repaired. Animals were sacrificed at 4, 8, and 12 weeks after the operation, and the knee that had the ligament repair was compared with the contralateral control knee. All knees were tested manually tested for stability to valgus stress and then prepared for histologic examination. Medial collateral ligaments repaired using the biodegradable suture anchor demonstrated stability to valgus stress and anatomic healing at the bone-tendon junction. Resorption of the implant was virtually complete by 12 weeks. All specimens demonstrated less inflammatory reaction to the suture anchor than to the attached Vicryl suture. This contrasts with the control group, which was grossly unstable and demonstrated scarring in this nonanatomic position. These results demonstrate efficacy of this particular material of biodegradable implant and justify further investigative efforts.

Video-assisted thoracic corpectomy and spinal reconstruction: a biomechanical analysis of open versus endoscopic technique.

The purpose of this study is to evaluate the technique of video-assisted thoracic surgery (VATS) to perform corpectomy and spinal reconstruction with the Harrington rod and polymethylmethacrylate construct, and to compare the biomechanical stability of the constructs created with both open thoracotomy and VATS technique. Fourteen farm-raised pigs were divided into two groups. Group I underwent thoracic corpectomy using a VATS approach and group II had the same procedure performed through a traditional thoracotomy. The stiffness (newtons per millimeter) for the non-destructive tests for each loading mode are flexion-compression (open 124.4 +/- 124.9; VAT 75.8 + 29.9); extension-compression (open 165.8 + 41.8: VATS 96.5 + 31.2); pure compression (open 231.4 + 126.4; VATS 264.6 + 184.3). The difference between group I (VATS) and group II (open) in extension compression is statistically significant. Although not statistically significant, the results also show the group II (open) specimens to be stiffer in flexion-compression and the group I (VATS) specimens to be stiffer in pure compression. Although corpectomy and spinal reconstruction can be performed with the VATS technique, the constructs obtained endoscopically may not have the same strength as those constructs obtained via an open procedure.

Use of stents covered with polytetrafluoroethylene in experimental abdominal aortic aneurysm.

PURPOSE: To establish the effectiveness of covered stents in the treatment of aortic aneurysms, to investigate the histopathologic healing patterns of the device, and to determine the long-term endurance and integrity of modified polytetrafluoroethylene (PTFE). MATERIALS AND METHODS: Experimental aneurysms were created in dogs by enlarging the aortic lumen with a patch of abdominal fascia. After 5 months, eight animals underwent an endoluminal bypass. The bypass device consisted of a 6-cm-long stent covered with thin PTFE. After surgery, the animals were killed at 3, 6, and 12 months in groups of three, three, and two, respectively. Specimens were processed for luminal surface studies and cross-sectional histologic study. Explanted PTFE material was analyzed for its physical characteristics and performance and was compared with retained control samples. RESULTS: Before the animals were killed, aortography showed patent bypass conduits in all animals, although two of eight had leaks into the aneurysmal sac. Endothelialized neointima largely covered the luminal surface of the PTFE stent. The percentage of prosthetic surface covered by tissue did not change from 3 months to 1 year. Physical testing of the explanted PTFE material showed no structural deterioration and no change in the internodal distance. Thickness and axial tensile strength varied 12% and 17% from controls, respectively. CONCLUSION: Thin-walled PTFE seems to have physicochemical characteristics that make this material adequate for endovascular use. Though limited, this study supports the establishment of preliminary clinical evaluation of metallic stents combined with PTFE for the treatment of abdominal aortic aneurysm.

Laparoscopic aortofemoral bypass. Initial experience in an animal model.

OBJECTIVE: The study objective was to evaluate the feasibility of laparoscopic aortofemoral bypass in a porcine model. SUMMARY BACKGROUND DATA: Laparoscopic techniques have been applied to numerous general and thoracic surgical procedures. Their application to vascular surgery has been virtually nonexistent. Open surgery for aortoiliac occlusive disease is accompanied by significant morbidity rates, and minimally invasive procedures have the disadvantage of reduced patency rates. Laparoscopic aortofemoral replacement has the theoretical advantage of long-term patency with reduced postoperative complications. METHODS: Between January and September 1993, laparoscopic surgery was performed on 16 pigs: 6 underwent transperitoneal laparoscopic aortic dissection and vessel control alone; 7 underwent complete transperitoneal laparoscopic aortofemoral bypass; and 3 underwent a retroperitoneal approach. The aortic anastomosis was performed using a combination of sutures and titanium clips in an end-to-side fashion in five pigs, and a custom-made nonsutured graft was secured with use of an end-to-end method in five pigs. Femoral anastomoses were performed with the standard open technique. RESULTS: Technical success was achieved in all 10 animals and with no major complications. Mean blood loss was 20 ml (range, 5-50 ml), and mean operative time was 2.45 hours (range, 2-4 hrs). On aortic-clamp release, 2 of the end-to-side anastomoses required additional sutures to stop bleeding between oversized staples, and 2 of the end-to-end anastomoses required additional ties to reinforce loose ties. All 10 grafts and anastomoses were patent and free of leaks after completion of the procedure. CONCLUSIONS: Laparoscopic aortofemoral bypass is technically feasible in a porcine model. Further experimental work with new instrumentation and technical refinement will make laparoscopic surgery feasible for the treatment of vascular disease in humans.

Biomechanical properties of hip cartilage in experimental animal models.

The material properties of normal adult articular cartilage were determined in the femoral head and acetabulum of baboons, dogs, and bovines, and were compared with those of normal human hip cartilage. In situ creep and recovery indentation experiments were performed using an automated creep indentation apparatus. To curvefit the entire creep curve, a numerical algorithm based on biphasic finite element methods and nonlinear optimization was developed. This effort represents the first successful use of 100% of the creep indentation curve to obtain the mechanical properties of normal articular cartilage. The results show that material properties of articular cartilage exhibit significant topographical variations in the femoral head and acetabulum, and between these two bone structures. Furthermore, significant differences exist in the mechanical properties of hip cartilage among the 4 species. Specifically, in all species the smallest aggregate modulus is found in the inferior aspect of the femoral head. Among all species, human hip cartilage is the stiffest in all test sites; bovine tissue is the softest. Human tissue has the smallest Poisson's ratio and permeability in all test sites. The aggregate modulus of human hip cartilage is closely resembled by that of baboon hip cartilage. Anatomically, canine and baboon hips exhibit similar characteristics to the human hip joint; the bovine hip joint is distinctly different. Based on this study's data, the baboon represents the most appropriate animal model of normal human hip articular cartilage.

Laparoscopic intracorporeal harvest of jejunal tissue for autologous transplantation.

Our goal in this study was to assess the feasibility and safety of performing an intracorporeal laparoscopic jejunal harvest. The initial technique was developed and refined in a pig and then a dog model. In the animal studies, careful dissection of the jejunal flap with its feeding vessel was accomplished along with an intracorporeal anastomosis. The laparoscopic dissection was facilitated by temporarily anchoring the jejunal flap to the anterior abdominal wall and transilluminating the mesentery. We present the first case report of a patient who underwent a laparoscopic jejunal harvest, intracorporeal small bowel anastomosis, and a microvascular anastomosis in the neck for reconstruction of the laryngopharynx.

Vascular occlusion with a balloon-expandable stent occluder.

PURPOSE: To evaluate the effectiveness of a new vascular occlusion device. MATERIALS AND METHODS: The device was created by coating a balloon-expandable stent with a silicone sleeve that tapers to a blind-ended nozzle at its leading end. Once crimp-mounted on an angioplasty balloon catheter, the device is introduced over a guide wire through a small end hole in the nozzle. RESULTS: The device was tested for stability and occlusive ability. No migration was measured over a pulsatile pulse range of 50-300 mm Hg, and mean flow rate in the occluded vessel was reduced from 443 mL/min +/- 99 (standard deviation) to 1.9 mL/min +/- 2.7. Subsequently, 12 arteries were occluded in three dogs, and immediate vascular occlusion was achieved in all vessels. An arteriovenous fistula was created in another six dogs and was successfully occluded with the device. Follow-up arteriography at 3 months demonstrated persistent occlusion with no migration of the device. CONCLUSION: This new occlusive device offers immediate vascular occlusion with excellent stability.

An initial comparative study of two techniques of laparoscopic colonic anastomosis and mesenteric defect closure.

Laparoscopic colon and rectal surgery is still in its nascent stages of development. The ease, efficacy, and safety of intracorporeal mechanical colonic anastomosis are contingent upon expensive stapling devices. Although mobilization and mesenteric division are feasible, a method of inexpensive rapid anastomosis is not. A single inexpensive multifire stapler which could be used both to fashion the anastomosis and to close the mesenteric defect would be ideal. Therefore, this prospective randomized study was undertaken to compare the clinical and functional results of laparoscopic colotomy closure performed using the Endopath EMS hernia stapler (EMS; Ethicon Endosurgery Inc., Cincinnati, OH) to results of using standard two-layer hand suturing (HS). Both the colotomy itself and the mesenteric defect closure sites were included in the randomization and analysis. The abdominal cavity was assessed for evidence of anastomotic leakage, abscess, and adhesion formation. In addition, radiographic luminal diameter, bursting strength, and histology were evaluated. Eight healthy pigs were randomized to either the EMS (N = 4) or HS (N = 4). There was no evidence of leakage, abscesses, or adhesion formation in either group; however, the mesenteric defect revealed more scarring in the HS than in the EMS animals. There were no significant differences in either luminal diameter (HS: mean = 0.92 cm; EMS: mean = 0.91 cm) or bursting strength (HS: mean = 171 mm Hg; EMS: mean = 157 mm Hg) (P > 0.05). Histologic analysis also demonstrated no difference in inflammation, necrosis, or fibrosis. This study suggests that this technique can be safely applied to both colotomy closure and mesenteric defect repair.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of synchronous and asynchronous pulsatile flow during left, right, and biventricular bypass.

Ventricular assist devices augment flow from the left atrium to the aorta and/or from the right atrium to the pulmonary artery. Most devices are used in the asynchronous full-to-empty mode (asynchronous) but may also be used in a synchronous counterpulsation mode (synchronous). This study determines the optimal assist modes to reduce myocardial oxygen consumption (MVO2) and metabolism. Twelve pigs were instrumented with carotid artery and Baim coronary sinus catheters for determination of MVO2 and myocardial lactate production (LACT). Six were implanted with a Pierce-Donachy left ventricular assist device (LVAD) and 6 with both right and left ventricular assist devices (BIVAD). Two periods each of control, synchronous, and asynchronous bypass were instituted, the midanterior descending coronary artery (LAD) was ligated, and the sequence was repeated. After each period, MVO2 and LACT were determined and myocardial biopsy specimens were obtained for tissue, lactate, and ATP assay. Following LAD ligation, biopsy specimens were obtained from both the infarct and noninfarct zones of the heart. MVO2 decreased (p < 0.05) in the asynchronous BIVAD mode compared with control. MVO2 was unchanged in synchronous BIVAD or either LVAD mode. Tissue ATP and tissue lactate were unaffected by any mode of bypass. Only BIVAD in the asynchronous mode reduced MVO2. When ventricular assist devices are utilized to aid recovery of the natural heart, two devices should always be inserted to allow biventricular assist. Synchronous counterpulsation offers no advantage.

Intraluminal bypass of abdominal aortic aneurysm: feasibility study.

This study evaluated the feasibility of treatment of abdominal aortic aneurysm (AAA) by means of intraluminal bypass. Artificial aortic aneurysms with fusiform Dacron conduits were created at surgery , a weft-knit Dacron tube with balloon-expandable stents attached at both ends was inserted transfemorally through a 14-F introducer sheath and expanded at the aneurysmal level by means of inflation of a coaxial balloon. The transluminal bypass successfully excluded the aneurysmal lumen immediately after placement. In two dogs, a postplacement aortogram revealed torsion of the graft that caused early occlusion. The six other dogs had a patent bypass at 6 months follow-up. Four of these dogs had evidence of kinking of the graft as a result of shrinkage of the prosthetic aneurysm. Histopathologic studies revealed that endothelialization was complete on the stents and partial on the graft surface. It is concluded that transluminal bypass of AAA is feasible in the dog model.

Brachygnathia in horses: 20 cases (1979-1989).

The medical records of 20 horses admitted to the veterinary medical center with a diagnosis of brachygnathia over a 10-year period (1979 to 1989) were reviewed. The study included 18 foals and 2 adult horses. Males were affected 5.7 times more frequently than females. The amount of disparity between the mandible and premaxilla varied between 0.75 and 3 cm. Sixteen foals were treated surgically with the temporary application of premaxillary tension band devices. Thirteen of the 16 surgical cases were available for follow-up evaluation. All of the surgically treated animals had improved incisive occlusion, and 6 foals had complete resolution of the deformity with corrections ranging from 0.75 to 2.5 cm. Complete correction of the malocclusion was more likely to occur if foals were treated when they were less than or equal to 6 months old. The average amount of correction achieved in foals treated when they were less than or equal to 6 months old was 1.5 cm. (range, 0.75 to 2.5 cm). Foals treated, when they were 7 to 12 months old, had an average of 0.6 cm of reduction in the malocclusion (range, 0.25 to 1 cm). Implant failure was the most common complication and occurred in 9 of the 13 foals treated surgically.

Effect of synchronous and asynchronous pulsatile flow during left, right, and biventricular bypass.

Ventricular assist devices augment aortic or pulmonary flow while the patient's heart recovers from surgery or infarction. Most are used in the asynchronous full-to-empty mode, but they also may be used in a synchronous counter-pulsation mode. This study examines which assist mode optimally reduces myocardial oxygen consumption (MVO2). Eighteen pigs were instrumented with pulmonary artery, carotid artery, and coronary sinus catheters for determination of MVO2. Pierce-Donachy Ventricular Assist Devices (VAD) were used in left, right, or biventricular assist mode. Fifteen minute periods each of control, synchronous, and asynchronous bypass were randomly instituted. The mid-left anterior descending coronary artery was then ligated, and the sequence repeated. At the end of each period, MVO2 was determined. In comparison with controls, MVO2 was statistically significant in the BIVAD asynchronous mode only. Synchronized counterpulsation did not decrease MVO2. When ventricular assist devices are used to aid in cardiac recovery postoperatively or postmyocardial infarction, biventricular assist should be used.

Transfixation pinning and casting of tibial fractures in calves: five cases (1985-1989).

Medical records of 5 calves with tibial fractures that were reduced and stabilized by transfixation pinning and casting were reviewed. Multiple Steinmann pins were placed transversely through proximal and distal fracture fragments, and the pin ends were incorporated in fiberglass cast material after fracture reduction. Cast material serves as an external frame to maintain pin position and fracture reduction. Calves were between 2 weeks and 6 months old and weighed between 40 and 180 kg. Three fractures were spiral in configuration and 2 were comminuted. One tibial fracture was open. After surgery, all calves were ambulatory within 24 hours. To improve tarsal flexion and achieve normal stance in 3 calves, cast revision was required on the caudal aspect of the limb. Good radiographic and clinical evidence of stability was observed in 5 to 10 weeks (mean 8 weeks), at which time the pis and cast were removed. Return to normal function was rapid and judged to be excellent at follow-up evaluation 3 to 12 months later. Advantages of transfixation pinning and casting in management of tibial fractures include flexibility in pin positioning, adequate maintenance of reduction, early return to weight-bearing status, joint mobility, and ease of ambulation. The inability to adjust fixation and alignment after cast application is a disadvantage of this technique compared with other external fixators.

Induction of peritoneal adhesions with small intestinal ischaemia and distention in the foal.

Twenty-two foals were divided into groups of intestinal distension and intestinal ischaemia as methods to induce peritoneal adhesions. In the first group, the lumen of a segment of distal small intestine was occluded without extramural vascular compromise and distended with lactated Ringer's solution to a constant pressure of 25 cm H2O for 2 h within the abdomen. The ischaemic group underwent 70 mins total vascular occlusion of identical segments of bowel. Serosal biopsies were obtained before and after each experimental procedure and following 60 mins of reperfusion. Similar biopsies were harvested from a control group of foals with no bowel occlusions. The foals were destroyed 10 days after surgery and tissues collected for histological and ultrastructural evaluation. Experimental and control mesothelial surfaces were denuded histologically immediately after experimental occlusions. Serosal oedema and cellular infiltration were observed following reperfusion of the ischaemic segments but were present immediately after 2 h of distension. All foals had developed bowel-to-bowel and bowel-to-mesentery adhesions of the experimental segments. Control foals under 30 days old exhibited mesenteric contraction and thickening of the isolated segment whereas those older than 30 days had little or no mesenteric thickening or contraction. Histologically, in the experimental segments, fibrous tissue had formed on the outer boundary of the original serosa, and new mesothelial-like cells were present on the surface of fibrous tissue in some areas. Some serosal fibrosis was also seen in most of the control segments.

The effects of fixation of the ulna to the radius in young foals.

The effects of radioulnar fixation were studied in 21 Quarter horse foals by applying a bone plate to the caudal aspect of the proximal part of the ulna, with screws engaging both the radius and the ulna. The plates were applied at 1 month of age in six foals (group I), 5 months of age in six foals (group II), and 7 months of age in three foals (group III). Six foals underwent sham operations at 1 month of age to serve as controls (group IV). Ulnar dysplasia and elbow subluxation developed in all treated foals. The magnitude of ulnar dysplasia was inversely related to the patient's age at fixation and was accompanied by degenerative joint disease and lameness in foals undergoing fixation at 1 and 5 months of age. Removal of the fixation appliances 16 weeks after implantation in three foals from each of groups I and II failed to reverse the degree of ulnar dysplasia. Although foals undergoing fixation at 7 months of age (group III) were not lame, radiographic evidence of subluxation and subtle degenerative changes in the articular cartilage of the treated elbow did develop. Recommendations for avoidance of radioulnar fixation were developed from these observations.

Osseous sequestration in the horse. A review of 68 cases.

Sixty-eight cases of cortical bone sequestration in 67 equine patients were reviewed with regard to the clinical presentation, method of treatment, and outcome. All lesions were located in skeletal areas with minimal soft tissue coverage, with 53% of them in the metatarsal and metacarpal bones. At the time of admission, 60% of the patients with limb lesions were lame; the majority improved with therapy. After sequestrectomy, there was a trend for surgical wounds which could be managed by primary closure to heal more rapidly (3.8 weeks) than wounds which required second intention healing (6.4 weeks). The appearance of healed wounds, as evaluated by owners, was acceptable in 45 cases. Dissatisfaction with the appearance of the healed wounds in 22 cases suggested a need to address the issue of cosmetic results.

Rectal leiomyosarcoma in a horse.

A mass originating from the dorsal rectal wall of a 4-year-old mare was diagnosed as a leiomyosarcoma. General anesthesia and meticulous surgical dissection were required to effect removal. In spite of incomplete excision, as indicated by histologic sections, the mass has not recurred in the 18 months since removal.