Despite our fine-grain anatomical knowledge of the cerebellar cortex, electrophysiological studies of circuit information processing over the last fifty years have been hampered by the difficulty of reliably assigning signals to identified cell types. We approached this problem by assessing the spontaneous activity signatures of identified cerebellar cortical neurones. A range of statistics describing firing frequency and irregularity were then used, individually and in combination, to build Gaussian Process Classifiers (GPC) leading to a probabilistic classification of each neurone type and the computation of equi-probable decision boundaries between cell classes. Firing frequency statistics were useful for separating Purkinje cells from granular layer units, whilst firing irregularity measures proved most useful for distinguishing cells within granular layer cell classes. Considered as single statistics, we achieved classification accuracies of 72.5% and 92.7% for granular layer and molecular layer units respectively. Combining statistics to form twin-variate GPC models substantially improved classification accuracies with the combination of mean spike frequency and log-interval entropy offering classification accuracies of 92.7% and 99.2% for our molecular and granular layer models, respectively. A cross-species comparison was performed, using data drawn from anaesthetised mice and decerebrate cats, where our models offered 80% and 100% classification accuracy. We then used our models to assess non-identified data from awake monkeys and rabbits in order to highlight subsets of neurones with the greatest degree of similarity to identified cell classes. In this way, our GPC-based approach for tentatively identifying neurones from their spontaneous activity signatures, in the absence of an established ground-truth, nonetheless affords the experimenter a statistically robust means of grouping cells with properties matching known cell classes. Our approach therefore may have broad application to a variety of future cerebellar cortical investigations, particularly in awake animals where opportunities for definitive cell identification are limited.
Hemodynamics constitute a critical factor in the formation of intracranial aneurysms. However, little is known about how intracranial arteries respond to hemodynamic insult and how that response contributes to aneurysm formation. We examined early cellular responses at rabbit basilar termini exposed to hemodynamic insult that initiates aneurysmal remodeling.
Flow in the basilar artery was increased by bilateral carotid artery ligation. After 2 and 5 days, basilar terminus tissue was examined by immunohistochemistry and quantitative PCR.
Within 2 days of flow increase, internal elastic lamina (IEL) was lost in the periapical region of the bifurcation, which experienced high wall shear stress and positive wall shear stress gradient. Overlying endothelium was still largely present in this region. IEL loss was associated with localized apoptosis and elevated expression of matrix metalloproteinases (MMPs) 2 and 9. A small number of inflammatory cells were sporadically scattered in the bifurcation adventitia and were not concentrated in regions of IEL loss and MMP elevation. Elevated MMP expression colocalized with smooth muscle α-actin in the media.
The initial vascular response to aneurysm-initiating hemodynamic insult includes localized matrix degradation and cell apoptosis. Such destructive remodeling arises from intrinsic mural cells, rather than through inflammatory cell infiltration.
Intracranial aneurysm; Hemodynamic forces; Vascular remodeling; Internal elastic lamina; Matrix metalloproteinase; Apoptosis; Inflammatory cells; Wall shear stress
Homonymous quadrantanopsia results from retrochiasmal lesions in the visual pathway. Invasive mole is a benign tumor that arises from myometrial invasion of a hydatidiform mole via direct extension through tissue or venous channels. Cerebral metastasis of invasive mole is rare and there has been no report demonstrating homonymous quadrantanopsia as the first manifestation of metastasis in any trophoblastic neoplasms.
We report the case of a 31-year-old Asian woman who presented with right homonymous inferior quadrantanopsia from the mass effect of a solitary cerebral metastasis from an invasive mole. A magnetic resonance image (MRI) of the brain showed a metastatic tumor in the left occipital lobe. The visual field improved slightly after chemotherapy. There was a reduction in the tumor size and the surrounding edema. This is the first case report demonstrating that homonymous quadrantanopsia should be included in the manifestations of the metastasis of an invasive mole.
The presentation of homonymous quadrantanopsia must alert ophthalmologists to conduct a complete medical history and arrange specialist consultation.
Cerebral metastasis; Homonymous Quadrantanopsia; Invasive mole
miR-34a functions as an important tumor suppressor during the process of carcinogenesis. However, the mechanism of miR-34a dysregulation in human malignancies has not been well elucidated. Our study aimed to further investigate the regulation mechanism of miR-34a.
We found that overexpression of NF-kappa B p65 subunit could increase miR-34a levels in EC109, an esophageal squamous cancer cell line, while ectopic expression of DN IkappaB leaded to a significant reduction of miR-34a expression. Bioinformatics analysis suggested three putative KB sites in promoter region of miR-34a gene. Mutation two of these KB sites impaired p65 induced miR-34a transcriptional activity. Chromatin immunoprecipitation and electrophoretic mobility shift assays both showed that NF-kappaB could specifically bind to the third KB site located in miR-34a promoter. In addition, we found that overexpression of NF-kappaB p65 could not successfully induce miR-34a expression in esophageal cancer cell lines with mutant p53 or decreased p53. Reporter assay further showed that NF-kappaB-induced miR-34a transcriptional activity was reduced by p53 impairment. Nevertheless, CHIP analysis suggested binding of NF-kappaB to miR-34a promoter was not affected in cells with mutant p53.
Our work indicates a novel mechanism of miR-34a regulation that NF-kappaB could elevate miR-34a expression levels through directly binding to its promoter. And wildtype p53 is responsible for NF-kappaB-mediated miR-34a transcriptional activity but not for NF-kappaB binding. These findings might be helpful in understanding miR-34a abnormality in human malignancies and open new perspectives for the roles of miR-34a and NF-kappaB in tumor progression.
miR-34a; NF-kappa B; p53; gene expression regulation
Background and Purpose
To identify significant morphologic and hemodynamic parameters that discriminate intracranial aneurysm (IA) rupture status using 3D angiography and computational fluid dynamics (CFD).
119 IAs (38 ruptured, 81 unruptured) were analyzed from 3D angiographic images and CFD. Six morphologic and seven hemodynamic parameters were evaluated for significance with respect to rupture. Receiver-operating characteristic (ROC) analysis identified area under the curve (AUC) and optimal thresholds separating ruptured from unruptured aneurysms for each parameter. Significant parameters were examined by multivariate logistic regression analysis in 3 predictive models—morphology only, hemodynamics only, and combined—to identify independent discriminants, and the AUC-ROC of the predicted probability of rupture status was compared among these models.
Morphologic parameters (Size Ratio [SR], Undulation Index, Ellipticity Index, and Nonsphericity Index) and hemodynamic parameters (Average Wall Shear Stress [WSS], Maximum intra-aneurysmal WSS, Low WSS Area, Average Oscillatory Shear Index [OSI], Number of Vortices, and Relative Resident Time) achieved statistical significance (p<0.01). Multivariate logistic regression analysis demonstrated SR to be the only independently significant factor in the morphology model (AUC=0.83, 95% confidence interval [CI] 0.75–0.91), whereas WSS and OSI were the only independently significant variables in the hemodynamics model (AUC=0.85, 95% CI 0.78–0.93). The combined model retained all three variables, SR, WSS, and OSI (AUC=0.89, 95% CI 0.82–0.96).
All three models—morphological (based on SR), hemodynamic (based on WSS and OSI), and combined—discriminate IA rupture status with high AUC values. Hemodynamics is as important as morphology in discriminating aneurysm rupture status.
Hemodynamics; Intracranial aneurysm; Morphology; Rupture
Cyclin D1 overexpression is a common feature of many human malignancies. Genomic deletion analysis has demonstrated a key role for cyclin D1 in cellular proliferation, angiogenesis and cellular migration. To investigate the mechanisms contributing to cyclin D1 functions, we purified cyclin D1a-associated complexes by affinity chromatography and identified the PACSIN 2 (protein kinase C and casein kinase substrate in neurons 2) protein by mass spectrometry. The PACSIN 2, but not the related PACSIN 1 and 3, directly bound wild-type cyclin D1 (cyclin D1a) at the carboxyl terminus and failed to bind cyclin D1b, the alternative splicing variant of cyclin D1. PACSIN 2 knockdown induced cellular migration and reduced cell spreading in LNCaP cells expressing cyclin D1a. In cyclin D1−/− mouse embryonic fibroblasts (MEFs), cyclin D1a, but not cyclin D1b, reduced the cell spreading to a polarized morphology. siPACSIN 2 had no effect on cellular migration of cyclin D1−/− MEFs. Cyclin D1a restored the migratory ability of cyclin D1−/− MEFs, which was further enhanced by knocking down PACSIN 2 with siRNA. The cyclin D1-associated protein, PACSIN 2, regulates cell spreading and migration, which are dependent on cyclin D1 expression.
PACSIN 2; cyclin D1; polymorphism; cellular migration; cell spreading; cancer
Lysophospholipids such as lysophosphatidic acid (LPA) and sphingosine-1-phosphate (S1P) are important signaling molecules that can regulate a wide range of cellular responses. We discovered that Sphingosine kinase 1 (Sphk1), a key enzyme that converts sphingosine to S1P, is expressed in neurons and progenitor cells in nascent trigeminal and dorsal root ganglia during mouse embryogenesis.
Methods and Findings
Sphk1 null mouse embryos do not display overt deficits owing to compensation by Sphk2. Thus, we analyzed embryos that are deficient in both Sphk1 and Sphk2 (which essentially eliminates S1P function) in order to investigate the role(s) of Sphk1 during sensory ganglia formation. While animals lacking 1–3 alleles of Sphk1 and Sphk2 had no obvious phenotype, embryos without both genes displayed clear developmental defects. The complete absence of Sphk1 and Sphk2 resulted in trigeminal and dorsal root ganglia with fewer neurons and progenitor cells. The profound loss in cell number could be attributed to a decrease in cell proliferation as well as an increase in apoptosis. Furthermore, Sphk1/2 double mutants displayed an overall reduction in other sphingolipids as well as an imbalance of S1P/sphingosine and S1P/ceramide ratio, thereby favoring cell death and reducing cell growth.
Together, these results provide strong in vivo evidence that sphingosine kinase/S1P signaling plays an important role in regulating early events during development of sensory ganglia.
Background and Purpose
Hemodynamic insult by bilateral common carotid artery (CCA) ligation has been shown to induce aneurysmal remodeling at the basilar terminus in a rabbit model. To characterize critical hemodynamics that initiate this remodeling, we applied a novel hemodynamics-histology co-mapping technique.
Eight rabbits received bilateral CCA ligation to increase basilar artery flow. Three underwent sham operations. Hemodynamic insult at the basilar terminus was assessed by computational fluid dynamics. Bifurcation tissue was harvested on day 5; histology was co-mapped with initial postligation hemodynamic fields of wall shear stress (WSS) and WSS gradient (WSSG).
All bifurcations showed internal elastic lamina (IEL) loss in periapical regions exposed to accelerating flow with high WSS and positive WSSG. IEL damage happened 100% of the time at locations where WSS>122 Pa and WSSG>530 Pa/mm. The degree of destructive remodeling accounting for IEL loss, medial thinning, and luminal bulging correlated with the magnitude of the hemodynamic insult.
Aneurysmal remodeling initiates when local hemodynamic forces exceed specific limits at the rabbit basilar terminus. A combination of high WSS and positive WSSG represents “dangerous” hemodynamics likely to induce aneurysmal remodeling.
Destructive remodeling; regional blood flow; hemodynamics; imaging; mapping
Flow diversion is a novel concept for intracranial aneurysm treatment. The recently developed Enterprise Vascular Reconstruction Device (Codman Neurovascular, Raynham MA) provides easy delivery and repositioning. Although designed specifically for restraining coils within an aneurysm, this stent has theoretical effects on modifying flow dynamics, which have not been studied. The goal of this study was to quantify the effect of single and multiple self-expanding Enterprise stents alone or in combination with balloon-mounted stents on aneurysm hemodynamics using computational fluid dynamics (CFD).
The geometry of a wide-necked, saccular, basilar trunk aneurysm was reconstructed from computed tomographic angiography images. Various combinations of 1–3 stents were “virtually” conformed to fit into the vessel lumen and placed across the aneurysm orifice. CFD analysis was performed to calculate hemodynamic parameters considered important in aneurysm pathogenesis and thrombosis for each model.
The complex aneurysmal flow pattern was suppressed by stenting. Stent placement lowered average flow velocity in the aneurysm; further reduction was achieved by additional stent deployment. Aneurysmal flow turnover time, an indicator of stasis, was increased to 114-117% for single-stent, 127-128% for double-stent, and 141% for triple-stent deployment. Furthermore, aneurysmal wall shear stress (WSS) decreased with increasing number of deployed stents.
This is the first study analyzing flow modifications associated with placement of Enterprise stents for aneurysm occlusion. Placement of 2-3 stents significantly reduced intra-aneurysmal hemodynamic activities, thereby increasing the likelihood of inducing aneurysm thrombotic occlusion.
Aneurysm; computational fluid dynamics; flow diversion; rupture; stent; turnover time
An asymmetric stent with low porosity patch across the intracranial aneurysm neck and high porosity elsewhere is designed to modify the flow to result in thrombogenesis and occlusion of the aneurysm and yet to reduce the possibility of also occluding adjacent perforator vessels. The purposes of this study are to evaluate the flow field induced by an asymmetric stent using both numerical and digital subtraction angiography (DSA) methods and to quantify the flow dynamics of an asymmetric stent in an in vivo aneurysm model. We created a vein-pouch aneurysm model on the canine carotid artery. An asymmetric stent was implanted at the aneurysm, with 25% porosity across the aneurysm neck and 80% porosity elsewhere. The aneurysm geometry, before and after stent implantation, was acquired using cone beam CT and reconstructed for computational fluid dynamics (CFD) analysis. Both steady-state and pulsatile flow conditions using the measured waveforms from the aneurysm model were studied. To reduce computational costs, we modeled the asymmetric stent effect by specifying a pressure drop over the layer across the aneurysm orifice where the low porosity patch was located. From the CFD results, we found the asymmetric stent reduced the inflow into the aneurysm by 51%, and appeared to create a stasis-like environment which favors thrombus formation. The DSA sequences also showed substantial flow reduction into the aneurysm. Asymmetric stents may be a viable image guided intervention for treating intracranial aneurysms with desired flow modification features.
Stent; Aneurysm; Digital Subtraction Angiography; Computational Fluid Dynamics; Image guided interventions; CT; image-based finite element models of physiology; image-based biomechanical models; circulation; porosity; wall shear stress
The phytohormone abscisic acid (ABA) and reactive oxygen species (ROS) play critical roles in mediating abiotic stress responses in plants. It is well known that ABA is involved in the modulation of ROS levels by regulating ROS-producing and ROS-scavenging genes, but the molecular mechanisms underlying this regulation are poorly understood. Here we show that the expression of maize ABP9 gene, which encodes a bZIP transcription factor capable of binding to the ABRE2 motif in the maize Cat1 promoter, is induced by ABA, H2O2, drought and salt. Constitutive expression of ABP9 in transgenic Arabidopsis leads to remarkably enhanced tolerance to multiple stresses including drought, high salt, freezing temperature and oxidative stresses. ABP9 expressing Arabidopsis plants also exhibit increased sensitivity to exogenously applied ABA during seed germination, root growth and stomatal closure and improved water-conserving capacity. Moreover, constitutive expression of ABP9 causes reduced cellular levels of ROS, alleviated oxidative damage and reduced cell death, accompanied by elevated expression of many stress/ABA responsive genes including those for scavenging and regulating ROS. Taken together, these results suggest that ABP9 may play a pivotal role in plant tolerance to abiotic stresses by fine tuning ABA signaling and control of ROS accumulation.
Electronic supplementary material
The online version of this article (doi:10.1007/s11103-011-9732-x) contains supplementary material, which is available to authorized users.
Transcription factor ABP9; ABA; Reactive oxygen species; Stress tolerance; Gene expression
Stenting may provide a new, less invasive therapeutic option for cerebral aneurysms. However, a conventional porous stent may be insufficient in modifying the blood flow for clinical aneurysms. We designed an asymmetric stent consisting of a low porosity patch welded onto a porous stent for an anterior cerebral artery aneurysm of a specific patient geometry to block the strong inflow jet. To evaluate the effect of the patch on aneurysmal flow dynamics, we “virtually” implanted it into the patient's aneurysm geometry and performed Computational Fluid Dynamics (CFD) analysis. The patch was computationally deformed to fit into the vessel lumen segmented from the patient CT reconstructions. After the flow calculations, a patch with the same design was fabricated using laser cutting techniques and welded onto a commercial porous stent, creating a patient-specific asymmetric stent. This stent was implanted into a phantom, which was imaged with X-ray angiography. The hemodynamics of untreated and stented aneurysms were compared both computationally and experimentally. It was found from CFD of the patient aneurysm that the asymmetric stent effectively blocked the strong inflow jet into the aneurysm and eliminated the flow impingement on the aneurysm wall at the dome. The impact zone with elevated wall shear stress was eliminated, the aneurysmal flow activity was substantially reduced, and the flow was considerably reduced. Experimental observations corresponded well qualitatively with the CFD results. The demonstrated asymmetric stent could lead to a new minimally invasive image guided intervention to reduce aneurysm growth and rupture.
Stent; Aneurysm; Computational Fluid Dynamics; Image guided interventions; Angiography; Asymmetric stent
In the title compound, C15H26O3·H2O, a sesquiterpenoid molecule with a germacrene backbone that contains two epoxide groups and one hydroxyl group. Intermolecular O—H⋯O hydrogen bonds between the epoxy groups and solvent water molecules give rise to an infinite three-dimensional supramolecular structure.
The effectiveness of intracranial aneurysm (IA) size as a predictor for rupture has been debated. We recently performed a retrospective analysis of IA morphology and found that a new index, namely, aneurysm-to-parent vessel size ratio (SR), was strongly correlated with IA rupture, with 77% of ruptured IAs showing an SR of more than 2, and 83% of unruptured IAs showing an SR of 2 or less. As hemodynamics have been implicated in both IA development and rupture, we examine how varying SR influences intra-aneurysmal hemodynamics.
One sidewall and 1 terminal IA were virtually reconstructed from patient 3-dimensional angiographic images. In 2 independent in silico experiments, the SR was varied from 1.0 to 3.5 by virtually changing either aneurysm size or vessel diameter while keeping the other parameter constant. Pulsatile computational fluid dynamics simulations were performed on each model for hemodynamics analysis.
Low SR (≤2) aneurysm morphology consistently demonstrated simple flow patterns with a single intra-aneurysmal vortex, whereas higher SR (>2) aneurysm morphology presented multiple vortices and complex flow patterns. The aneurysm luminal area that was exposed to low wall shear stress increased with increasing SR. Complex flow, multiple vortices, and low aneurysmal wall shear stress have been associated with ruptured IAs in previous studies.
Higher SR, irrespective of aneurysm type and absolute aneurysm or vessel size, gives rise to flow patterns typically observed in ruptured IAs. These results provide hemodynamic support for the existing correlation of SR with rupture risk.
Intracranial aneurysm; Morphology; Parent vessel diameter; Rupture risk; Size ratio
Pathological extremes in cerebrovascular remodeling may contribute to basilar artery (BA) dolichoectasia and fusiform aneurysm development. Factors regulating cerebrovascular remodeling are poorly understood. To better understand hemodynamic influences on cerebrovascular remodeling, we examined BA remodeling following common carotid artery (CCA) ligation in an animal model.
Rabbits were subjected to sham surgery (3 animals), unilateral CCA ligation (3 animals), or bilateral CCA ligation (5 animals). Transcranial Doppler ultrasonography and rotational angiography were used to compute BA flow, diameter, wall shear stress (WSS), and a tortuosity index on Days 0, 1, 4, 7, 14, 28, 56, and 84. Basilar artery tissues were stained and analyzed at Day 84. Statistical analysis was performed using orthogonal contrast analysis, repeated measures analysis of variance, or mixed regression analysis of repeated measures. Statistical significance was defined as a probability value < 0.05.
Basilar artery flow and diameter increased significantly after the procedure in both ligation groups, but only the bilateral CCA ligation group demonstrated significant differences between groups. Wall shear stress significantly increased only in animals in the bilateral CCA ligation group and returned to baseline by Day 28, with 52% of WSS correction occurring by Day 7. Only the bilateral CCA ligation group developed significant BA tortuosity, occurring within 7 days postligation. Unlike the animals in the sham and unilateral CCA ligation groups, the animals in the bilateral CCA ligation group had histological staining results showing a substantial internal elastic lamina fragmentation.
Increased BA flow results in adaptive BA remodeling until WSS returns to physiological baseline levels. Morphological changes occur rapidly following flow alteration and do not require chronic insult to affect substantial and significant structural transformation. Additionally, it appears that there exists a flow-increase threshold that, when surpassed, results in significant tortuosity.
basilar artery; carotid artery occlusion; dolichoectasia; rabbit; tortuosity; wall shear stress
The porous intravascular stents that are currently available may not cause complete aneurysm thrombosis and may therefore fail to provide durable protection against aneurysm rupture when used as a sole treatment modality. The goal of this study was to quantify the effects of porous stents on aneurysm hemodynamics using computational fluid dynamics.
The geometry of a wide-necked saccular basilar trunk aneurysm was reconstructed from a patient′s computed tomographic angiography images. Three commercial stents (Neuroform2; Boston Scientific/Target, San Leandro, CA; Wingspan; Boston Scientific, Fremont, CA; and Vision; Guidant Corp., Santa Clara, CA) were modeled. Various combinations of one to three stents were virtually conformed to fit into the vessel lumen and placed across the aneurysm orifice. An unstented aneurysm served as a control. Computational fluid dynamics analysis was performed to calculate the hemodynamic parameters considered important in aneurysm pathogenesis and thrombosis for each of the models.
The complex flow pattern observed in the unstented aneurysm was suppressed by stenting. Stent placement lowered the wall shear stress in the aneurysm, and this effect was increased by additional stent deployment. Turnover time was moderately increased after single- and double-stent placement and markedly increased after three stents were placed. The influence of stent design on hemodynamic parameters was more significant in double-stented models than in other models.
Aneurysm hemodynamic parameters were significantly modified by placement of multiple stents. Because the associated modifications may be helpful as well as harmful in terms of rupture risk, use of this technique requires careful consideration.
Aneurysm; Endovascular; Hemodynamics; Multiple stenting; Stent
Abnormal vascular remodeling mediated by inflammatory cells has been identified as a key pathologic component of various vascular diseases, including abdominal aortic aneurysms, brain arteriovenous malformations and atherosclerosis. Based on findings from observational studies that analysed human intracranial aneurysms and experimental studies that utilized animal models, an emerging concept suggests that a key component of the pathophysiology of intracranial aneurysms is sustained abnormal vascular remodeling coupled with inflammation. This concept may provide a new treatment strategy to utilize agents to inhibit inflammation or cytokines produced by inflammatory cells such as matrix metalloproteinases. Such an approach would aim to stabilize these vascular lesions and prevent future expansion or rupture.
Intracranial aneurysms; inflammation; remodeling; pathophysiology
Computational fluid dynamics (CFD) simulations using medical-image-based anatomical vascular geometry are now gaining clinical relevance. This study aimed at validating the CFD methodology for studying cerebral aneurysms by using particle image velocimetry (PIV) measurements, with a focus on the effects of small geometric variations in aneurysm models on the flow dynamics obtained with CFD. Method of Approach. An experimental phantom was fabricated out of silicone elastomer to best mimic a spherical aneurysm model. PIV measurements were obtained from the phantom and compared with the CFD results from an ideal spherical aneurysm model (S1). These measurements were also compared with CFD results, based on the geometry reconstructed from three-dimensional images of the experimental phantom. We further performed CFD analysis on two geometric variations, S2 and S3, of the phantom to investigate the effects of small geometric variations on the aneurysmal flow field. Results. We found poor agreement between the CFD results from the ideal spherical aneurysm model and the PIV measurements from the phantom, including inconsistent secondary flow patterns. The CFD results based on the actual phantom geometry, however, matched well with the PIV measurements. CFD of models S2 and S3 produced qualitatively similar flow fields to that of the phantom but quantitatively significant changes in key hemodynamic parameters such as vorticity, positive circulation, and wall shear stress. Conclusion. CFD simulation results can closely match experimental measurements as long as both are performed on the same model geometry. Small geometric variations on the aneurysm model can significantly alter the flow-field and key hemodynamic parameters. Since medical images are subjected to geometric uncertainties, image-based patient-specific CFD results must be carefully scrutinized before providing clinical feedback.
computational fluid dynamics; CFD validations; particle image velocimetry; aneurysm; circulation; hemodynamics; geometric uncertainties
Cerebral aneurysms are preferentially located at arterial bifurcation apices with complex hemodynamics. To understand disease mechanisms associated with aneurysm initiation, we attempted to establish a causal relationship between local hemodynamics and vascular responses.
Arterial bifurcations were surgically created from native common carotid arteries in two dogs, angiographically imaged 2 weeks and 2 months later, and then excised. We characterized local morphological changes in response to specifically manipulated hemodynamics. Computational fluid dynamics simulations were performed on the in vivo images and results mapped onto histological images.
Local flow conditions, such as high wall shear stress and high wall shear stress gradient, were found to be associated with vascular changes, including an intimal pad in the flow impingement region and a “groove” bearing the characteristics of an early aneurysm.
This novel method of histohemodynamic micromapping reveals a direct correlation between an altered hemodynamic microenvironment and vascular responses consistent with aneurysm development.
Arterial bifurcation; Cerebral aneurysms; Hemodynamics; Vascular remodeling
Although elevated hemodynamics has been speculated to play a key role in intracranial aneurysm (IA) initiation, little is known about the specific hemodynamic microenvironment that triggers aneurysmal vascular degradation. We previously demonstrated maladaptive remodeling characteristic of IA initiation occurring in hemodynamic regions of combined high wall shear stress (WSS) and high WSS gradient near the apex of an experimentally created carotid bifurcation. This study examines whether this remodeling recapitulates the molecular changes found in IAs and whether molecular changes also correspond to specific hemodynamic environments.
De novo bifurcations were surgically created using both native common carotid arteries in each of 6 dogs. Bifurcations were imaged 2 weeks or 2 months after surgery by high-resolution 3-dimensional angiography, from which flow fields were obtained by computational fluid dynamics simulations. Subsequently, harvested tissues, demonstrating early aneurysmal changes near the apex, were immunostained for interleukin-1β, endothelial and inducible nitric oxide synthases, nitrotyrosine, and matrix metalloproteinase-2 and -9. Spatial distributions of these molecules were comapped with computational fluid dynamics results.
The aneurysmal wall showed decreased endothelial nitric oxide synthase expression compared with surrounding segments, the feeding artery, and native controls, whereas all other markers increased. Anti-CD68 staining indicated the absence of inflammatory cells in the aneurysmal wall. Comapping molecular marker distributions with flow fields revealed confinement of these molecular changes within the hemodynamic region of high WSS and high, positive WSS gradient.
Aneurysm-initiating remodeling induced by combined high WSS and high, positive WSS gradient is associated with molecular changes implicated in IAs.
Aneurysm; Initiation; Matrix metalloproteinase; Nitric oxide; Pathogenesis; Wall shear stress
Background and Purpose
Arterial bifurcation apices are common sites for cerebral aneurysms, raising the possibility that the unique hemodynamic conditions associated with flow dividers predispose the apical vessel wall to aneurysm formation. This study sought to identify the specific hemodynamic insults that lead to maladaptive vascular remodeling associated with aneurysm development and to identify early remodeling events at the tissue and cellular levels.
We surgically created new branch points in the carotid vasculature of 6 female adult dogs. In vivo angiographic imaging and computational fluid dynamics simulations revealed the detailed hemodynamic microenvironment for each bifurcation, which were then spatially correlated with histologic features showing specific tissue responses.
We observed 2 distinct patterns of vessel wall remodeling: (1) hyperplasia that formed an intimal pad at the bifurcation apex and (2) destructive remodeling in the adjacent region of flow acceleration that resembled the initiation of an intracranial aneurysm, characterized by disruption of the internal elastic lamina, loss of medial smooth muscle cells, reduced proliferation of smooth muscle cells, and loss of fibronectin.
Strong localization of aneurysm-type remodeling to the region of accelerating flow suggests that a combination of high wall shear stress and a high gradient in wall shear stress represents a “dangerous” hemodynamic condition that predisposes the apical vessel wall to aneurysm formation.
wall shear stress; gradient; intimal hyperplasia; intracranial aneurysm
There is a general lack of quantitative understanding about how specific design features of endovascular stents (struts and mesh design, porosity) affect the hemodynamics in intracranial aneurysms. To shed light on this issue, we studied two commercial high-porosity stents (Tristar stent™ and Wallstent®) in aneurysm models of varying vessel curvature as well as in a patient-specific model using Computational Fluid Dynamics. We investigated how these stents modify hemodynamic parameters such as aneurysmal inflow rate, stasis, and wall shear stress, and how such changes are related to the specific designs. We found that the flow damping effect of stents and resulting aneurysmal stasis and wall shear stress are strongly influenced by stent porosity, strut design, and mesh hole shape. We also confirmed that the damping effect is significantly reduced at higher vessel curvatures, which indicates limited usefulness of high-porosity stents as a stand-alone treatment. Finally, we showed that the stasis-inducing performance of stents in 3D geometries can be predicted from the hydraulic resistance of their flat mesh screens. From this, we propose a methodology to cost-effectively compare different stent designs before running a full 3D simulation.
Stent design; Cerebral aneurysm; Vessel geometry; Anatomical aneurysm; Hemodynamics; Hydraulic resistance; Computational Fluid Dynamics; Tristar stent™; Wallstent®
Inverse and forward dynamic models have been conceptually important in computational motor control. In particular, inverse models are thought to convert desired action into appropriate motor commands. In parallel, forward models predict the consequences of the motor command on behavior by constructing an efference copy of the actual movement. Despite theoretical appeal, their neural representation has remained elusive. Here we provide evidence supporting the notion that a group of premotor neurons called ‘Burst-Tonic’ (BT) cells represent the output of the inverse model for eye movements. We show that BT neurons, like extraocular motoneurons but different from the evoked eye movement, do not carry signals appropriate for the ‘half-angle rule’ of ocular kinematics during smooth pursuit eye movements from eccentric positions. Along with findings of identical response dynamics as motoneurons, these results strongly suggest that BT cells carry a replica of the motor command. In contrast, Eye-Head (EH) neurons, a premotor cell type that is the target of Purkinje cell inhibition from the cerebellar flocculus/ventral paraflocculus, exhibit properties that could be consistent with the half-angle rule. Thus, EH cells may be functionally related to the output of a forward internal model thought to construct an efference copy of the actual eye movement.
torsion; listing’s law; half angle rule; burst tonic cells; internal model; smooth pursuit
The aim of this study is to identify image-based morphological parameters that correlate with human intracranial aneurysm (IA) rupture.
For 45 patients with terminal or sidewall saccular IAs (25 unruptured, 20 ruptured), three-dimensional geometries were evaluated for a range of morphological parameters. In addition to five previously studied parameters (aspect ratio, aneurysm size, ellipticity index, nonsphericity index, and undulation index), we defined three novel parameters incorporating the parent vessel geometry (vessel angle, aneurysm [inclination] angle, and [aneurysm-to-vessel] size ratio) and explored their correlation with aneurysm rupture. Parameters were analyzed with a two-tailed independent Student's t test for significance; significant parameters (P < 0.05) were further examined by multivariate logistic regression analysis. Additionally, receiver operating characteristic analyses were performed on each parameter.
Statistically significant differences were found between mean values in ruptured and unruptured groups for size ratio, undulation index, nonsphericity index, ellipticity index, aneurysm angle, and aspect ratio. Logistic regression analysis further revealed that size ratio (odds ratio, 1.41; 95% confidence interval, 1.03−1.92) and undulation index (odds ratio, 1.51; 95% confidence interval, 1.08−2.11) had the strongest independent correlation with ruptured IA. From the receiver operating characteristic analysis, size ratio and aneurysm angle had the highest area under the curve values of 0.83 and 0.85, respectively.
Size ratio and aneurysm angle are promising new morphological metrics for IA rupture risk assessment. Because these parameters account for vessel geometry, they may bridge the gap between morphological studies and more qualitative location-based studies.
Intracranial aneurysm; Morphology; Rupture risk; Size ratio; Vessel geometry
Little is known about endothelial responses to the impinging flow hemodynamics that occur at arterial bifurcation apices, where intracranial aneurysms usually form. Such hemodynamic environments are characterized by high wall shear stress (WSS >40 dynes/cm2) and high wall shear stress gradients (WSSG >300 dynes/cm3). In this study, confluent bovine aortic endothelial cells were exposed to impinging flow in a T-shaped chamber designed to mimic a bifurcation. After 24−72 h under flow, cells around the stagnation point maintained polygonal shapes but cell density was reduced, whereas cells in adjacent downstream regions exposed to very high WSS and WSSG were elongated, aligned parallel to flow, and at higher density. Such behavior was not blocked by inhibiting proliferation, indicating that cells migrated downstream from the stagnation point in response to impinging flow. Furthermore, although the area of highest cell density moved downstream and away from the impingement point over time, it never moved beyond the WSS maximum. The accumulation of cells upstream of maximal WSS and downstream of maximal WSSG suggests that positive WSSG is responsible for the observed migration. These results demonstrate a unique endothelial response to aneurysm-promoting flow environments at bifurcation apices.
Wall shear stress; Spatial wall shear stress gradient; Migration; Aneurysm