Stephen B. Lewis

Lessons learned from whole exome sequencing in multiplex families affected by a complex genetic disorder, intracranial aneurysm

Genetic risk factors for intracranial aneurysm (IA) are not yet fully understood. Genomewide association studies have been successful at identifying common variants; however, the role of rare variation in IA susceptibility has not been fully explored. In this study, we report the use of whole exome sequencing (WES) in seven densely-affected families (45 individuals) recruited as part of the Familial Intracranial Aneurysm study. WES variants were prioritized by functional prediction, frequency, predicted pathogenicity, and segregation within families. Using these criteria, 68 variants in 68 genes were prioritized across the seven families. Of the genes that were expressed in IA tissue, one gene (TMEM132B) was differentially expressed in aneurysmal samples (n=44) as compared to control samples (n=16) (false discovery rate adjusted p-value=0.023). We demonstrate that sequencing of densely affected families permits exploration of the role of rare variants in a relatively common disease such as IA, although there are important study design considerations for applying sequencing to complex disorders. In this study, we explore methods of WES variant prioritization, including the incorporation of unaffected individuals, multipoint linkage analysis, biological pathway information, and transcriptome profiling. Further studies are needed to validate and characterize the set of variants and genes identified in this study.

Treatment of giant intracranial aneurysms with saphenous vein extracranial-to-intracranial bypass grafting: Indications, operative technique, and results in 29 patients

OBJECTIVE The treatment of giant intracranial aneurysms is a challenge because of the limitations and difficulty of direct surgical clipping and endovascular coiling. We describe the indications, surgical technique, and complications of saphenous vein extracranial-to-intracranial bypass grafting followed by acute parent vessel occlusion in the management of these difficult lesions. METHODS Between January 1990 and December 1999, 29 patients with giant intracranial aneurysms underwent 30 saphenous vein bypass grafts followed by immediate parent vessel occlusion. There were 11 men and 18 women with a mean follow-up period of 62 months. Twenty-five patients harbored aneurysms involving the internal carotid artery, 2 had middle cerebral artery aneurysms, and 2 had aneurysms in the basilar artery. Serial cerebral or magnetic resonance angiograms were obtained to assess graft patency and aneurysm obliteration. RESULTS All 30 aneurysms were excluded from the cerebral circulation, with 28 vein grafts remaining patent. Two patients had graft occlusions: one because of poor runoff and the other because of misplacement of a cranial pin during a bypass procedure on the contralateral side. Other surgical complications included one death from a large cerebral infarction, homonymous hemianopsia from thrombosis of an anterior choroidal artery after internal carotid artery occlusion, and temporary hemiparesis from a presumed perforator thrombosis adjacent to a basilar aneurysm. CONCLUSION With appropriate attention to surgical technique, a saphenous vein extracranial-to-intracranial bypass followed by acute parent vessel occlusion is a safe and effective method of treating giant intracranial aneurysms. A high rate of graft patency and adequate cerebral blood flow can be achieved. Thrombosis of perforating arteries caused by altered blood flow hemodynamics after parent vessel occlusion may be a continuing source of complications.

Cerebral oxygenation monitoring by near-infrared spectroscopy is not clinically useful in patients with severe closed-head injury: A comparison with jugular venous bulb oximetry

OBJECTIVEnTo compare continuous jugular venous bulb oximetry and cerebral near-infrared spectroscopy in patients with severe closed head injury.nnnDESIGNnA prospective observational study.nnnSETTINGnIntensive care unit of a major teaching hospital.nnnPATIENTSnAdults (n = 10) with severe closed-head injury (Glasgow Coma Scale score of < or = 8).nnnINTERVENTIONSnNone.nnnMEASUREMENTS AND MAIN RESULTSnJugular venous bulb oximetry, cerebral near-infrared spectroscopy, and cerebral perfusion pressure were measured continuously. A total of 3,691 paired measurements of near-infrared spectroscopy and jugular venous bulb oximetry were analyzed. Poor correlation (r2 = .04) between paired measurements and wide limits of agreement (-13% to +21%) were demonstrated. The mean difference between measurements was +/- 4% and the standard deviation of the mean difference was +/- 8.69%. The data were subsequently grouped according to three clinically significant subgroups of jugular venous bulb oxygen saturation reflecting low ( < 55%), normal (55% to 75%) and high ( > 75%) saturation values. Poor correlation and wide limits of agreement between the two methods of measurement were observed in all groups. Values recorded by near-infrared spectroscopy did not significantly change between the groups, and 14 clinically significant episodes of jugular venous bulb desaturation were not detected by near-infrared spectroscopy.nnnCONCLUSIONSnTissue oxygen saturation determined by near-infrared spectroscopy does not reflect significant changes in cerebral oxygenation detected by the global measurement of jugular venous bulb oximetry. This finding may be explained by inadequate signal detection and inaccuracies in the algorithm used to filter out extracranial components. Until these technical difficulties are addressed, near-infrared spectroscopy, as measured by the machine assessed in this study, cannot be routinely recommended for assessment of cerebral oxygenation in patients with acute head injury.

Bottleneck factor and height-width ratio: association with ruptured aneurysms in patients with multiple cerebral aneurysms.

OBJECTIVEDetermining factors predictive of the natural risk of rupture of cerebral aneurysms is difficult because of the need to control for confounding variables. We studied factors associated with rupture in a study model of patients with multiple cerebral aneurysms, one aneurysm that had ruptured and one or more that had not, in which each patient served as their own internal control. METHODSWe collected aneurysm location, one-dimensional measurements, and two-dimensional indices from the computed tomographic angiograms of patients in the proposed study model and compared ruptured versus unruptured aneurysms. Bivariate statistics were supplemented with multivariable logistic regression analysis to model ruptured status. A total of 40 candidate models were evaluated for predictive power and fit with Wald scoring, Cox and Snell R2, Hosmer and Lemeshow tests, case classification counting, and residual analysis to determine which of the computed tomographic angiographic measurements or indices were jointly associated with and predictive of aneurysm rupture. RESULTSThirty patients with 67 aneurysms (30 ruptured, 37 unruptured) were studied. Maximum diameter, height, maximum width, bulge height, parent artery diameter, aspect ratio, bottleneck factor, and aneurysm/parent artery ratio were significantly (P < 0.05) associated with ruptured aneurysms on bivariate analysis. When best subsets and stepwise multivariable logistic regression was performed, bottleneck factor (odds ratio = 1.25, confidence interval = 1.11–1.41 for every 0.1 increase) and height-width ratio (odds ratio = 1.23, confidence interval = 1.03–1.47 for every 0.1 increase) were the only measures that were significantly predictive of rupture. CONCLUSIONIn a case-control study of patients with multiple cerebral aneurysms, increased bottleneck factor and height-width ratio were consistently associated with rupture.

αII-Spectrin Breakdown Product Cerebrospinal Fluid Exposure Metrics Suggest Differences in Cellular Injury Mechanisms after Severe Traumatic Brain Injury

Traumatic brain injury (TBI) produces alphaII-spectrin breakdown products (SBDPs) that are potential biomarkers for TBI. To further understand these biomarkers, the present study examined (1) the exposure and kinetic characteristics of SBDPs in cerebrospinal fluid (CSF) of adults with severe TBI, and (2) the relationship between these exposure and kinetic metrics and severity of injury. This clinical database study analyzed CSF concentrations of 150-, 145-, and 120-kDa SBDPs in 38 severe TBI patients. Area under the curve (AUC), mean residence time (MRT), maximum concentration (C(max)), time to maximum concentration (T(max)), and half-life (t(1/2)) were determined for each SBDP. Markers of calpain proteolysis (SBDP150 and SBDP145) had a greater median AUC and C(max) and a shorter MRT than SBDP120, produced by caspase-3 proteolysis in the CSF in TBI patients ( p < 0.001). AUC and MRT for SBDP150 and SBDP15 were significantly greater in patients with worse Glasgow Coma Scale (GCS) scores at 24 h after injury compared to those whose GCS scores improved (AUC p=0.013, MRT p=0.001; AUC p=0.009, MRT p=0.021, respectively). A positive correlation was found between patients with longer elevations in intracranial pressure (ICP) measurements of 25mmHg or higher and those with a greater AUC and MRT for all three biomarkers. This is the first study to show that the biomarkers of proteolysis differentially associated with calpain and caspase-3 activity have distinct CSF exposure profiles following TBI that suggest a prominent role for calpain activity. Further studies are being conducted to determine if exposure and kinetic metrics for biofluid-based biomarkers can predict clinical outcome.

Proteomic identification of biomarkers of traumatic brain injury.

Traumatic brain injury (TBI) is a major national health problem without a US Food and Drug Administration-approved therapy. This review summarizes the importance of discovering relevant TBI protein biomarkers and presents logical rationale that neuroproteomic technologies are uniquely suited for the discovery of otherwise unnoticed TBI biomarkers. It highlights that one must make careful decisions when choosing which paradigm (human vs. animal models) and which biologic samples to use for such proteomic studies. It further outlines some of the desirable attributes of an ideal TBI biomarker and discusses how biomarkers discovered proteomically are complementary to those identified by traditional approaches. Lastly, the most important sequela of any proteomically identified TBI biomarker is validation in preclinical or clinical samples.

THE EFFECT OF COILING VERSUS CLIPPING OF RUPTURED AND UNRUPTURED CEREBRAL ANEURYSMS ON LENGTH OF STAY, HOSPITAL COST, HOSPITAL REIMBURSEMENT, AND SURGEON REIMBURSEMENT AT THE UNIVERSITY OF FLORIDA

OBJECTIVEThere are few studies comparing the economic costs and reimbursements for aneurysm clipping versus coiling, and none are from the United States. Our hypothesis predicted that coiling would result in shorter lengths of hospitalization than clipping in patients with unruptured aneurysms and would therefore result in lower hospital charges. However, because of the severity of subarachnoid hemorrhage, there would be no difference in length of hospitalization or hospital charges in patients with ruptured aneurysms. METHODSWe compared aneurysm coiling with aneurysm clipping in patients with unruptured and ruptured aneurysms treated at the University of Florida from January 2005 to June 2007 for differences in length of hospitalization, hospital costs, hospital collections, and surgeon collections. Patient demographic and aneurysm characteristic data were obtained from a clinical database. Length of hospitalization, cost, billing, and collection data were obtained from the hospital cost accounting database. Multivariate statistical analyses of length of hospitalization, hospital costs, hospital collections, and surgeon collections were performed using factors including patient age, sex, aneurysm size, aneurysm location, aneurysm treatment, presence of subarachnoid hemorrhage, clinical grade, payor, hospital billing, and surgeon billing. RESULTSThere were 565 patients with cerebral aneurysms treated either surgically (306 patients, 54%) or endovascularly (259 patients, 46%). In patients without subarachnoid hemorrhage (unruptured aneurysms) (n = 367), surgery, compared with endovascular treatment, was associated with longer hospitalization (P < 0.001), but lower hospital costs (P < 0.001), higher surgeon collections (P = 0.003), and similar hospital collections. In patients with subarachnoid hemorrhage (ruptured aneurysms) (n = 198), surgery was associated with lower hospital costs (P = 0.011), but similar length of stay, surgeon collections, and hospital collections. Larger aneurysm size was significantly associated with longer hospitalization in the patients with unruptured aneurysms (P < 0.001) and higher hospital costs for both patients with unruptured (P < 0.001) and ruptured (P = 0.015) aneurysms. The payor was significantly associated with hospital costs in patients with ruptured aneurysms (P = 0.034) and length of stay (unruptured aneurysms, P < 0.001; ruptured aneurysms, P < 0.001), hospital collections (unruptured aneurysms, P < 0.001; ruptured aneurysms, P < 0.001), and surgeon collections (unruptured aneurysms, P < 0.001; ruptured aneurysms, P < 0.001) in both patients with unruptured and ruptured aneurysms. A worse clinical grade was significantly associated with higher hospital costs (P < 0.001). CONCLUSIONDespite a shorter length of hospitalization in patients with unruptured aneurysms, coiling was associated with higher hospital costs in both patients with unruptured and ruptured aneurysms. This is likely attributable to the higher device cost of coils than clips. The advantages of coiling over clipping would be better realized if the cost of coils could be comparably reduced to that of clips.

SKULL BASE CHORDOMA

Our purpose was to discuss the optimal treatment and outcomes for patients with skull base chordomas.

Detection of phosphorylated NF-H in the cerebrospinal fluid and blood of aneurysmal subarachnoid hemorrhage patients.

Blood and cerebrospinal fluid (CSF) of 30 Fisher grade 3 aneurysmal subarachnoid hemorrhage (ASAH) patients were analyzed for the presence of the phosphorylated axonal form of the major neurofilament subunit NF-H (pNF-H), a promising biomarker of axonal injury. Patient demographic data including development of vasospasm and outcome scores at 6 months after aneurysmal rupture (AR) were evaluated. Higher pNF-H blood levels in the first few days after AR were strongly predictive of a negative outcome. Blood pNF-H levels in most recovering patients showed a steady increase into the second week after AR, presumably reflecting axonal degeneration secondary to the original insult. Almost half of the patients studied showed sudden dramatic peaks of pNF-H protein release into CSF in the 3- to 14-day time period after AR, which must reflect profound, coordinated, and secondary loss of axons. Patients in whom vasospasm was detected had significantly more pNF-H in both blood and CSF compared with those in whom vasospasm was not detected. We conclude that the analysis of pNF-H levels in blood and CSF differentiates between patients with poor and favorable outcomes and also reveals several novel features of ASAH progression and recovery.

Comparison of N-butyl cyanoacrylate and onyx for the embolization of intracranial arteriovenous malformations: analysis of fluoroscopy and procedure times.

OBJECTIVE Intracranial arteriovenous malformations (AVM) may be managed through staged preoperative embolization and resection. Two commonly used liquid embolics are N-butyl cyanoacrylate (nBCA; Cordis Microvascular, Inc., New Brunswick, NJ) and Onyx (ev3, Inc., Irvine, CA). We sought to compare the utility of these agents in terms of fluoroscopy and procedure times. METHODS All intracranial AVMs embolized from 2002 to 2006 at the University of Florida were included in this study. Patients were stratified into three treatment groups: nBCA, Onyx, and patients who received both nBCA and Onyx during separate embolizations. Cohorts were compared by sex, age, Spetzler-Martin grade, AVM volume, fluoroscopy time, procedure time, surgical blood loss, and complications. RESULTS A total of 182 embolizations were performed on 88 patients (nBCA, 60 patients and 106 procedures; Onyx, 20 patients and 43 procedures; and nBCA/Onyx, eight patients and 16 nBCA and 17 Onyx procedures). There were no significant differences in patient demographics, AVM volumes, and Spetzler-Martin grades. Mean fluoroscopy and procedure times were increased for Onyx (57 min; 2.6 h) compared with nBCA (37 min; 2.1 h) embolizations (P < 0.0001 and P = 0.001, respectively). Cumulative mean fluoroscopy time was increased for Onyx (135 min) and nBCA/Onyx (180 min) cohorts relative to nBCA (64 min; P < 0.0001). Cumulative mean procedure time was increased in the nBCA/Onyx group (10.4 h) compared with nBCA (3.7 h) and Onyx (5.4 h; P < 0.0001). Seventy patients (80%) underwent AVM resection. No significant differences in surgical blood loss or complication rates were observed among the cohorts. CONCLUSION Onyx AVM embolization requires increased fluoroscopy and procedure times compared with nBCA. Further investigation is necessary to justify increased radiation exposure and procedure time associated with Onyx.