Linda M. Graham

Relationship of age, gender, race, and body size to infrarenal aortic diameter

PURPOSE To assess the effects of age, gender, race, and body size on infrarenal aortic diameter (IAD) and to determine expected values for IAD on the basis of these factors. METHODS Veterans aged 50 to 79 years at 15 Department of Veterans Affairs medical centers were invited to undergo ultrasound measurement of IAD and complete a pre-screening questionnaire. We report here on 69,905 subjects who had no previous history of abdominal aortic aneurysm (AAA) and no ultrasound evidence of AAA (defined as IAD > or = 3.0 cm). RESULTS Although age, gender, black race, height, weight, body mass index, and body surface area were associated with IAD by multivariate linear regression (all p < 0.001), the effects were small. Female sex was associated with a 0.14 cm reduction in IAD and black race with a 0.01 cm increase in IAD. A 0.1 cm change in IAD was associated with large changes in the independent variables: 29 years in age, 19 cm or 40 cm in height, 35 kg in weight, 11 kg/m2 in body mass index, and 0.35 m2 in body surface area. Nearly all height-weight groups were within 0.1 cm of the gender means, and the unadjusted gender means differed by only 0.23 cm. The variation among medical centers had more influence on IAD than did the combination of age, gender, race, and body size. CONCLUSIONS Age, gender, race, and body size have statistically significant but small effects on IAD. Use of these parameters to define AAA may not offer sufficient advantage over simpler definitions (such as an IAD > or = 3.0 cm) to be warranted.

Celiac artery aneurysms: Historic (1745-1949) versus contemporary (1950-1984) differences in etiology and clinical importance

Celiac artery aneurysms were encountered in nine patients, ranging in age from 39 to 76 years, at the University of Michigan Medical Center between 1961 and 1983. Developmental defects and atherosclerosis were etiologic factors in six cases. Four patients were without symptoms, whereas five experienced abdominal pain, including one with a ruptured aneurysm. Eight patients were subjected to surgical treatment; no deaths occurred and symptoms were resolved in all patients. A literature review of 108 celiac artery aneurysms revealed two distinct subgroups. Among 60 celiac artery aneurysms encountered before 1950, representing the historic era, 40% were infectious (usually luetic), 7% were traumatic, and 52% were of undetermined cause. Most were symptomatic, 87% ruptured, and 95% were diagnosed at postmortem examination. The contemporary era since 1950 consisted of 48 cases, including nine in the Michigan experience. Congenital or developmental medial defects of the arterial wall and atherosclerosis were the most common causes of aneurysms. Most aneurysms in the contemporary period were either asymptomatic or accompanied by vague abdominal discomfort. Rupture affected 13% of those aneurysms. Operative therapy was successfully undertaken in 91% of 43 patients during the contemporary era, including eight in the present series.

Atherosclerotic extracranial carotid artery aneurysms.

Twenty-four atherosclerotic extracranial carotid artery aneurysms were encountered in 21 patients during a 25-year period. These represented 46% of all extracranial carotid artery aneurysms diagnosed at the University of Michigan during this period. Neurologic symptoms including amaurosis fugax, transient ischemic attacks, and stroke were present in 50% of the patients. An asymptomatic pulsatile neck mass occurred in 33%. Surgical therapy was undertaken for 18 aneurysms, and nonoperative treatment was pursued in the remaining six aneurysms. Operative therapy included 14 aneurysmectomies and four aneurysmorraphies. There were no surgical deaths. Transient perioperative neurologic deficits affected three of these patients (17%), and one individual (5%) experienced a permanent deficit. Transient cranial nerve deficits occurred in three patients (17%), and a permanent deficit was noted in one patient (5%). During a 7.6-year follow-up period no late strokes occurred among patients who were operated on. Nonoperative therapy was associated with three ipsilateral strokes during a mean follow-up period of 6.3 years. Atherosclerotic extracranial carotid artery aneurysms were associated with an exceptionally high stroke rate (50%) if treated nonoperatively. Prevention of late stroke justifies surgery, although perioperative neurologic deficits may accompany this therapy more often than with nonatherosclerotic carotid artery aneurysms.

Sequential studies of healing in endothelial seeded vascular prostheses histologic and ultrastructure characteristics of graft incorporation

Abstract Chronological events leading to incorporation of endothelial cell seeded prosthetic vascular grafts were documented in this investigation. Forty-one adult dogs underwent thoracoabdominal bypass using double-velour Dacron grafts. Experimental grafts were preclotted with blood containing enzymatically derived endothelium immediately after derivation, or after 14 days of cultivation. Control grafts were preclotted without addition of endothelial cells. Grafts were studied grossly as well as by light, scanning electron, transmission electron, and fluorescence microscopy, 1 to 28 days postimplantation. Control graft healing proceeded from pannus and perigraft ingrowth. Experimental grafts healed from seeded cells as well. Platelets covered all grafts by Days 1 and 2. Thrombus accumulations on control grafts, first evident on Day 4, became maximal by Day 14. Seeded grafts appeared relatively thrombus free with patches of endothelial cells noted by 4 days. These cells were initially separated by gaps, often containing leukocytes. Endothelium became densely packed with cellular migration and proliferation. Subendothelial tissues were composed of fibrin and smooth muscle. Control and experimental grafts were approximately 10 and 80% endothelialized, respectively, by Day 28. Smooth muscle dominated subintimal tissue in experimental grafts. These cells initially appeared fibroblastoid. Endothelial seeding enhances both pseudointimal development and rapid graft incorporation.

Elucidation of a TRPC6-TRPC5 Channel Cascade That Restricts Endothelial Cell Movement

Canonical transient receptor potential (TRPC) channels are opened by classical signal transduction events initiated by receptor activation or depletion of intracellular calcium stores. Here, we report a novel mechanism for opening TRPC channels in which TRPC6 activation initiates a cascade resulting in TRPC5 translocation. When endothelial cells (ECs) are incubated in lysophosphatidylcholine (lysoPC), rapid translocation of TRPC6 initiates calcium influx that results in externalization of TRPC5. Activation of this TRPC6-5 cascade causes a prolonged increase in intracellular calcium concentration ([Ca(2+)](i)) that inhibits EC movement. When TRPC5 is down-regulated with siRNA, the lysoPC-induced rise in [Ca(2+)](i) is shortened and the inhibition of EC migration is lessened. When TRPC6 is down-regulated or EC from TRPC6(-/-) mice are studied, lysoPC has minimal effect on [Ca(2+)](i) and EC migration. In addition, TRPC5 is not externalized in response to lysoPC, supporting the dependence of TRPC5 translocation on the opening of TRPC6 channels. Activation of this novel TRPC channel cascade by lysoPC, resulting in the inhibition of EC migration, could adversely impact on EC healing in atherosclerotic arteries where lysoPC is abundant.

Membrane microviscosity regulates endothelial cell motility

Endothelial cell (EC) movement is an initiating and rate-limiting event in the neogenesis and repair of blood vessels. Here, we explore the hypothesis that microviscosity of the plasma membrane (PM) is a key physiological regulator of cell movement. Aortic ECs treated with membrane-active agents, such as α-tocopherol, cholesterol and lysophospholipids, exhibited a biphasic dependency on membrane microviscosity, in which moderate increases enhanced EC migration, but increases beyond a threshold markedly inhibited migration. Surprisingly, angiogenic growth factors, that is, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), also increased membrane microviscosity, as measured in live cells by fluorescence recovery after photobleaching (FRAP). The localization of Rac to the PM was modified in cells treated with membrane-active agents or growth factors, suggesting a molecular mechanism for how membrane microviscosity influences cell movement. Our data show that angiogenic growth factors, as well as certain lipophilic molecules, regulate cell motility through alterations in membrane properties and the consequent relocalization of critical signalling molecules to membranes.

Spatial Coordination of Actin Polymerization and ILK–Akt2 Activity during Endothelial Cell Migration

Eukaryotic cell migration proceeds by cycles of protrusion, adhesion, and contraction, regulated by actin polymerization, focal adhesion assembly, and matrix degradation. However, mechanisms coordinating these processes remain largely unknown. Here, we show that local regulation of thymosin-beta4 (Tbeta4) binding to actin monomer (G-actin) coordinates actin polymerization with metalloproteinase synthesis to promote endothelial cell motility. In particular and quite unexpectedly, FRET analysis reveals diminished interaction between Tbeta4 and G-actin at the cell leading edge despite their colocalization there. Profilin-dependent dissociation of G-actin-Tbeta4 complexes simultaneously liberates actin for filament assembly and facilitates Tbeta4 binding to integrin-linked kinase (ILK) in the lamellipodia. Tbeta4-ILK complexes then recruit and activate Akt2, resulting in matrix metalloproteinase-2 production. Thus, the actin-Tbeta4 complex constitutes a latent coordinating center for cell migratory behavior, allowing profilin to initiate a cascade of events at the leading edge that couples actin polymerization to matrix degradation.

Polarization of Plasma Membrane Microviscosity during Endothelial Cell Migration

Cell movement is characterized by anterior-posterior polarization of multiple cell structures. We show here that the plasma membrane is polarized in moving endothelial cells (EC); in particular, plasma membrane microviscosity (PMM) is increased at the cell leading edge. Our studies indicate that cholesterol has an important role in generation of this microviscosity gradient. In vitro studies using synthetic lipid vesicles show that membrane microviscosity has a substantial and biphasic influence on actin dynamics; a small amount of cholesterol increases actin-mediated vesicle deformation, whereas a large amount completely inhibits deformation. Experiments in migrating ECs confirm the important role of PMM on actin dynamics. Angiogenic growth factor-stimulated cells exhibit substantially increased membrane microviscosity at the cell front but, unexpectedly, show decreased rates of actin polymerization. Our results suggest that increased PMM in lamellipodia may permit more productive actin filament and meshwork formation, resulting in enhanced rates of cell movement.

Lysophosphatidylcholine Inhibits Endothelial Cell Migration by Increasing Intracellular Calcium and Activating Calpain

Objective—Endothelial cell (EC) migration, essential for reestablishing arterial integrity after vascular injury, is inhibited by oxidized LDL (oxLDL) and lysophosphatidylcholine (lysoPC) that are present in the arterial wall. We tested the hypothesis that a mechanism responsible for lysoPC-induced inhibition is increased intracellular free calcium concentration ([Ca2+]i). Methods and Results—LysoPC, at concentrations that inhibit in vitro EC migration to 35% of control, increased [Ca2+]i levels 3-fold. These effects of lysoPC were concentration dependent and reversible. LysoPC induced Ca2+ influx within 10 minutes, and [Ca2+]i remained elevated for 2 hours. The calcium ionophore A23187 also increased [Ca2+]i and inhibited EC migration. Chelators of intracellular Ca2+ (BAPTA/AM and EGTA/AM) and nonvoltage-sensitive channel blockers (lanthanum chloride and gadolinium chloride) blunted the lysoPC-induced [Ca2+]i rise and partially preserved EC migration. After lysoPC treatment, calpain, a calcium-dependent cysteine protease, was activated, and cytoskeletal changes occurred. Calpain inhibitors (calpastatin, MDL28170, and calpeptin) added before lysoPC prevented cytoskeletal protein cleavage and preserved EC migration at 60% of control levels. Conclusions—LysoPC increases [Ca2+]i. In turn, activating calpains that can alter the cytoskeleton are activated and EC migration is inhibited.

Developmental occlusive disease of the abdominal aorta and the splanchnic and renal arteries

Developmental occlusive disease of the abdominal aorta and the renal and splanchnic arteries represent an unusual vascular condition. When unrecognized or untreated this disease is associated with premature death, usually from severe secondary hypertension as a consequence of renovascular stenotic lesions. Strong circumstantial evidence indicates that developmental abnormalities occurring during the fetal union of the two dorsal aortae account for most of the occlusive lesions affecting the abdominal aorta and its visceral branches in these patients. Complete arteriographic studies are necessary to confirm and accurately delineate the disease process. Surgical treatment, which often encompasses complex vascular reconstructive efforts, affords excellent results when carefully planned and executed.