Purification and characterization of biotin-binding protein II from chicken oocytes.

BBP-II, the major biotin-binding protein from chicken oocytes, was purified 12,000-fold with a 22% yield. The purification procedure includes butan-1-ol extraction of yolk lipids, phosphocellulose chromatography of the water-soluble proteins, DEAE-cellulose chromatography at pH 7.4 and hydroxyapatite column chromatography. Final purification was obtained by using a second DEAE-cellulose column chromatography at pH 6.0. BBP-I activity separated from BBP-II activity during elution from the first DEAE-cellulose column. Purified BBP-II was homogeneous on both polyacrylamide-gel electrophoresis and SDS/polyacrylamide-gel electrophoresis under conditions that would detect a 1% impurity. The subunit Mr determined from SDS/polyacrylamide-gel electrophoresis was 18,200 (72,600 for tetramer), which compares favourably with an Mr value of 17,300 (69,100) calculated from the amino acid analysis. A single precipitin line formed when rabbit antiserum to the protein was directed against a crude chicken egg-yolk sample. BBP-II purified by this procedure lacked carbohydrate and phosphate, was stable indefinitely when frozen, and was quite stable at room temperature. The N-terminal amino acid sequence showed polymorphism at three positions in the first 23 residues and was about 45% identical with the N-terminal 22 residues of avidin. Antiserum to BBP-II cross-reacted with BBP-I and similar proteins in the yolk of eggs from various birds and alligator as judged by immunodiffusion and enzyme-linked immunosorbent assays. No cross-reaction was observed with chicken egg-white by either of these methods.

Competitive binding assays for high-affinity binders in the presence of endogenous ligands: application to biotin-binding proteins.

Endogenous ligands complicate radioligand-binding assays of high-affinity binding proteins by obscuring binding sites or by diluting the labeled ligand. We have developed a mathematical model for such systems where radioligand and endogenous ligand are structurally identical. Data which relate radioligand binding at equilibrium as a function of sample volume can be plotted such that the concentrations of endogenous ligand and binder are graphically determined; however, a more precise determination may be done by nonlinear regression with the aid of a microcomputer. The method is demonstrated for the assay of biotin-binding proteins in the presence of a range of endogenous biotin concentrations below and above that required to saturate the binding sites.

Endoluminal repair of atypical dissecting aneurysm of descending thoracic aorta and fusiform aneurysm of the abdominal aorta.

A 62-year-old male patient was admitted with acute dissociation of the descending thoracic aorta and an infrarenal abdominal aortic aneurysm (AAA). Investigation revealed that the thoracic dissection probably had arisen retrogradely in the posterior wall of the AAA and extended superiorly to the left subclavian artery as a blind sac. Implantation of an endoluminal graft device below the renal arteries enabled simultaneous treatment of the AAA and the thoracic aortic dissection. The patient had an uncomplicated recovery. Postoperative aortography and computed tomography demonstrated normal flow through the aorta and endograft without leak of contrast into the AAA sac or the false lumen of the dissection. Contrast computed tomography 6 months after operation demonstrated that the false lumen was no longer evident.

Surgical management of complications following endoluminal grafting of abdominal aortic aneurysms.

OBJECTIVE: The aim of this study was to report the outcome of endoluminal grafting of abdominal aortic aneurysms (AAA) with special reference to complications. METHODS: Between May 1992 and August 1994 endoluminal repair of aneurysms was undertaken in 61 patients. In 53 the aneurysm was aortic and these are the basis of this report. In patients with AAA all procedures were elective and were performed in the operating room with the patient draped for an open repair in the event of failed endoluminal repair. The configuration of the endografts was tubular 36, tapered aortoiliac/aortofemoral 12 and bifurcated 5. Radiographic guidance was used to pass the endografts into the aorta via a delivery sheath introduced through the femoral or iliac arteries. RESULTS: Successful endoluminal repair of AAA was achieved in 43 of 53(81%) patients. In the remaining 10 patients, endoluminal repair was abandoned in favour of an open repair. There were 17(32%) local/vascular and 13(25%) systemic/remote remote complications. The sum of these complications occurring in successful endoluminal repairs and those complications leading to failure of endoluminal repair was 40(75%). There were two cardiac deaths within 30 days in patients undergoing endoluminal repair (both procedure related) and four late deaths (unrelated to aneurysm repair). Three of the late deaths were in patients undergoing endoluminal repair and one endoluminal converted to open repair. CONCLUSION: Endoluminal repair of AAA in our experience has a low perioperative (< 30 days) mortality rate (3.7%) but a high morbidity rate (75%). It is recommended that complications be classified into three groups: systemic/remote and local/vascular (following successful endoluminal repair) plus those complications leading to failure of endoluminal repair. The first group is composed of medical complications while the latter two groups comprise those surgical complications directly related to the endoluminal technique.

Results of autopsy 7 months after successful endoluminal treatment of an infrarenal abdominal aortic aneurysm.

PURPOSE: To report the results of a postmortem examination in a patient who died of unrelated causes 7 months following endoluminal treatment of an infrarenal abdominal aortic aneurysm (AAA). METHODS: As part of an FDA Phase I pilot study, a 73-year-old man underwent successful endoluminal exclusion of an infrarenal AAA using a 9-cm-long endograft (Endovascular Grafting System). Seven months later, he succumbed to complications of a spontaneous esophageal rupture. At autopsy, the aorta was dissected in situ by a vascular surgeon and pathologist before being explanted in order to examine the wound healing characteristics at the aorta-endograft interface. Particular attention was also directed to the hooks composing the attachment system at each end of the endograft. RESULTS: Macroscopic and microscopic examination revealed that the graft had completely excluded the aneurysm sac from the circulation and was incorporated into the aortic wall at the proximal neck and distal cuff. A smooth pannus of endothelial cells covered the proximal end of the endograft at the areas of contact with the aorta, while microscopic examination of the distal end of the graft revealed poorly formed, fibrinous pannus. The neointima deep to the endothelium consisted of a collagenous matrix containing myofibroblasts and histiocytes, providing evidence of healing between the endograft and aorta. Both renal arteries were clear of the proximal end of the endograft, but a previously unrecognized right lower pole renal artery with an extremely caudal origin was excluded from the aortic lumen. Each hook of the attachment system was seen protruding through the adventitia of the aorta. There was no evidence of trauma to the aortic wall or the surrounding tissues caused by these hooks. CONCLUSION: There appears to be evidence that an endoluminally placed aortic graft may be incorporated by the host aortic tissue.

Endoluminal grafting of abdominal aortic aneurysms: causes of failure and their prevention.

PURPOSE: The aim of this study was to analyze the causes of failure of endoluminal grafting for abdominal aortic aneurysms (AAA) and to put forward proposals for preventing these failures. METHODS: Since May 1992, endoluminal repair of aneurysms was undertaken in 47 patients. Forty-three of these patients had AAAs and are the basis of this study. All procedures were nonurgent and were performed in the operating room with the patient draped for an open repair in the event of failed endoluminal repair. Radiographic guidance was used to pass the endografts into the aorta via a delivery sheath introduced through the femoral or iliac arteries. The configuration of the endografts was tubular (n = 28), tapered aortoiliac/aortofemoral (n = 11), and bifurcated (n = 4). RESULTS: Successful endoluminal repair was achieved in 34 of 43 (79%) patients. The remaining nine were terminated in favor of an open repair. The causes of failure were problems with access (2), balloon malfunction (1), stent dislodgment (3), graft thrombosis (1), and inability to deploy the contralateral limb of a bifurcated graft (2). All failed endoluminal repairs proceeded to successful open repair. There was no perioperative mortality in patients undergoing endoluminal repair or in those whose endoluminal repair was converted to open operation. CONCLUSIONS: The failures of endoluminal grafting have been analyzed. Methods of avoiding access problems, balloon malfunction, and stent dislodgment have been defined and recommendations made.

Historic control comparison of outcome for matched groups of patients undergoing endoluminal versus open repair of abdominal aortic aneurysms.

PURPOSE: Currently no randomized studies show the relative morbidity and mortality of the open and endoluminal methods of abdominal aortic aneurysm (AAA) repair. The aim of this study was to analyze the outcome of two matched groups of patients with AAA, one undergoing open repair and the other undergoing endoluminal repair. METHODS: Two groups of patients who had undergone repair of AAA by open technique (group 1) or by endoluminal methods (group 2) were compared. A historic control cohort of 27 patients was selected from 56 consecutive patients who underwent open repair of AAA between January 1991 and February 1992. Patients considered unsuitable for the endoluminal method on the basis of computed tomography and aortography were excluded (n=29). Between May 1992 and November 1994 prospective data were recorded for 62 consecutive patients who underwent endoluminal repair by tube or bifurcated endografts. Twenty-eight patients who had been specifically referred for endoluminal AAA repairs because of preexisting severe medical comorbidities were excluded. Six of the endoluminal cases had failure, requiring conversion to open operation, and were excluded for separate analysis, leaving 28 patients in group 2. Patients in both groups were thus fit and suitable for either open or endoluminal repair and were comparable in relation to age, sex, risk factors, dimensions, and form of AAA. RESULTS: The mean values for operation time, blood loss, intensive care stay, and hospital stay for group 1 and group 2 were 2.6 versus 3.1 hours, 1422 versus 873 ml,* 1.8 versus 0.7 days,* and 12.4 versus 11.1 days, respectively (*p<0.05). Local/vascular complications occurred in 15% of patients in group 1 compared with 25% in group 2 (p=0.55), whereas remote/systemic complications occurred in 37% and 29%, respectively (p=0.3). Five of 28 patients in the endoluminal group had complications requiring early operative repair (n=3) or late revision (n=2). When comparison was made on an intention-to-treat basis (with failed procedures included), the incidence of local/vascular complications was significantly greater for endoluminal repair (p=0.047). CONCLUSIONS: The incidence of systemic/remote complications was similar for the two groups in spite of significantly less blood loss and shorter intensive care unit stay with endoluminal repair. The incidence of local/vascular complications had a tendency to be higher for endoluminal compared with standard open method (and was significantly greater if failed procedures were included). In this early experience with prototype devices, patients who were medically suitable for open surgical procedures did not derive benefit from the less invasive endoluminal technique with respect to duration of operation, length of hospital stay, or perioperative morbidity and mortality. On the other hand, because they also did not have worse outcome, a randomized study is now justified in this group.

Riboflavin-binding protein from reptiles: a comparison with avian riboflavin-binding proteins.

1. Riboflavin-binding protein (RBP) has been isolated for the first time from reptilian sources. 2. RBP from eggs of Python molurus (Indian python) and Chrysemys picta (painted turtle) has been isolated and compared to RBP from Gallus gallus domesticus (chicken), a well-characterized protein, and a newly isolated RBP from Cairina moschata (Muscovy duck). 3. Each of the proteins is phosphorylated and glycosylated. 4. The ratio of riboflavin binding to protein is 1:1 and the KD for each protein is between 1-3 nM. 5. The mol. wts, different for each species, range from 30,000-40,000, with the reptilian proteins being approx. 10,000 larger than the avian proteins.

Early experience with the Sydney and EVT prostheses for endoluminal treatment of abdominal aortic aneurysms.

PURPOSE: The aim of this study was to report early experiences with the Sydney and Endovascular Technologies (EVT) prostheses for the treatment of abdominal aortic aneurysms (AAA) deemed suitable for endoluminal tube graft repair. METHODS: Consecutive endoluminal tube graft repairs were analyzed over the first 12 months in which the Sydney and EVT prostheses were used. Patients eligible for the EVT prosthesis had type I AAAs: a proximal neck length > or = 2 cm, a distal cuff length > or = 1.5 cm, and nontortuous iliac arteries > or = 8 mm. Selection criteria for the Sydney device were more liberal and included AAAs that had distal cuffs < 1.5 cm. During the study period, 28 of 91 patients evaluated for AAA repair were thus selected for endoluminal grafting: 18 patients received the Sydney endograft and 10 the EVT device. Medical comorbidities were present in slightly less than one third of patients in both groups. Contrast-enhanced computerized tomography (CT) was performed preoperatively, within 10 days of operation, and at 6 and 12 months postprocedure. RESULTS: All endografts were successfully deployed in both groups. Postprocedural CT scans revealed incomplete aneurysm exclusion in four patients with the Sydney endograft. Subsequent deployment of a second endograft sealed these "leaks" in two cases; the other two were converted to open repair (89% clinical success). No leaks were seen with the EVT device. Local/vascular complications occurred in 33% of the Sydney group compared with 20% for the EVT device (p = 0.001); systemic sequelae were more common in the EVT group (30% versus 17% in the Sydney cohort, p = 0.002). There were no deaths within 30 days; three late deaths were not procedure related. CONCLUSION: AAAs that are suitable for endoluminal tube graft repair may be treated with a high rate of initial success with either the Sydney or EVT prostheses. More liberal selection criteria may increase the likelihood of local/vascular complications.