Hiroshi Tanemura

Mechanism of oxidative DNA damage induced by quercetin in the presence of Cu(II)

Quercetin, one of flavonoids, has been reported to be carcinogenic. There have been no report concerning carcinogenicity of kaempferol and luteolin which have structure similar to quercetin. DNA damage was examined by using DNA fragments obtained from the human p53 tumor suppressor gene. Quercetin induced extensive DNA damage via reacting with Cu(II), but kaempferol and luteolin induced little DNA damage even in the presence of Cu(II). Excessive quercetin inhibited copper-dependent DNA damage induced by quercetin. Bathocuproine, a Cu(I)-specific chelator, catalase and methional inhibited the DNA damage by quercetin, whereas free hydroxyl radical scavengers did not. Site specificity of the DNA damage was thymine and cytosine residues. The site specificity and the inhibitory effects suggested that DNA-copper-oxygen complex rather than free hydroxyl radical induced the DNA damage. Formation of 8-oxodG by quercetin increased extensively in the presence of Cu(II), whereas 8-oxodG formation by kaempferol or luteolin increased only slightly. This study suggests a good relationship between carcinogenicity and oxidative DNA damage of three flavonoids. The mechanism of DNA damage by quercetin was discussed in relation to the safety in cancer chemoprevention by flavonoids.

Low Wall Shear Stress Is Independently Associated With the Rupture Status of Middle Cerebral Artery Aneurysms

Background and Purpose— We determined which hemodynamic parameter independently characterizes the rupture status of middle cerebral artery (MCA) aneurysms using computational fluid dynamics analysis. Methods— In 106 patient-specific geometries of MCA aneurysms (43 ruptured, 63 unruptured), morphological and hemodynamic parameters were compared between the ruptured and unruptured groups. Multivariate logistic regression analysis was performed to determine parameters that independently characterized the rupture status of MCA aneurysms. Results— Univariate analyses showed that the aspect ratio, wall shear stress (WSS), normalized WSS, oscillatory shear index, WSS gradient, and aneurysm-formation index were significant parameters. The size of the aneurysmal dome and the gradient oscillatory number were not significantly different between the 2 groups. With multivariate analyses, only lower WSS was significantly associated with the rupture status of MCA aneurysms. Conclusions— WSS may be the most reliable parameter characterizing the rupture status of MCA aneurysms.

Inflammatory Biomarkers in Atherosclerosis: Pentraxin 3 Can Become a Novel Marker of Plaque Vulnerability

Inflammation is crucially involved in the development of carotid plaques. We examined the relationship between plaque vulnerability and inflammatory biomarkers using intraoperative blood and tissue specimens. We examined 58 patients with carotid stenosis. Following carotid plaque magnetic resonance imaging, 41 patients underwent carotid artery stenting (CAS) and 17 underwent carotid endarterectomy (CEA). Blood samples were obtained from the femoral artery (systemic) and common carotid artery immediately before and after CAS (local). Seventeen resected CEA tissue samples were embedded in paraffin, and histopathological and immunohistochemical analyses for IL-6, IL-10, E-selectin, adiponectin, and pentraxin 3 (PTX3) were performed. Serum levels of IL-6, IL-1β, IL-10, TNFα, E-selectin, VCAM-1, adiponectin, hs-CRP, and PTX3 were measured by multiplex bead array system and ELISA. CAS-treated patients were classified as stable plaques (n = 21) and vulnerable plaques (n = 20). The vulnerable group showed upregulation of the proinflammatory cytokines (IL-6 and TNFα), endothelial activation markers (E-selectin and VCAM-1), and inflammation markers (hs-CRP and PTX3) and downregulation of the anti-inflammatory markers (adiponectin and IL-10). PTX3 levels in both systemic and intracarotid samples before and after CAS were higher in the vulnerable group than in the stable group. Immunohistochemical analysis demonstrated that IL-6 was localized to inflammatory cells in the vulnerable plaques, and PTX3 was observed in the endothelial and perivascular cells. Our findings reveal that carotid plaque vulnerability is modulated by the upregulation and downregulation of proinflammatory and anti-inflammatory factors, respectively. PTX3 may thus be a potential predictive marker of plaque vulnerability.

High-Risk Plaque for Carotid Artery Stenting Evaluated With 3-Dimensional T1-Weighted Gradient Echo Sequence

Background and Purpose— Preventing cerebral embolisms is a major concern with carotid artery stenting (CAS). This study evaluated 3-dimensional T1-weighted gradient echo (3D T1GRE) sequence to predict cerebral embolism related to CAS. Methods— We performed quantitative analyses of the characteristics of 47 carotid plaques before CAS by measuring the signal intensity ratio (SIR) and plaque volume using 3D T1GRE images. We used T1-weighted turbo field echo sequence to obtain 3D T1GRE images. We also evaluated diffusion-weighted images (DWI) of the brain before and after CAS to detect ischemic lesions (DWI lesions) from cerebral emboli. Results— SIR (2.17 [interquartile range 1.50–3.07] versus 1.35 [interquartile range 1.08–1.97]; P=0.010) and plaque volume (456 mm3 [interquartile range 256–696] versus 301 mm3 [interquartile range 126–433]; P=0.008) were significantly higher in the group of patients positive for DWI lesions (P-group: n=26) than DWI lesion-negative patients (N-group: n=21). In multivariate logistic regression analysis, SIR (P=0.007) and plaque volume (P=0.042) were independent predictors of DWI lesions with CAS. Furthermore, SIR (rs=0.42, P=0.005) and plaque volume (rs=0.36, P=0.012) were positively correlated with the number of DWI lesions. From analysis of a receiver-operating characteristic curve, the most reliable cutoff values of SIR and plaque volume to predict DWI lesions related to CAS were 1.80 and 373 mm3, respectively. Conclusions— Quantitative evaluation of carotid plaques using 3D T1GRE images may be useful in predicting cerebral embolism related to CAS.

Stagnation and complex flow in ruptured cerebral aneurysms: a possible association with hemostatic pattern.

OBJECT Histopathological examination has revealed that ruptured cerebral aneurysms have different hemostatic patterns depending on the location of the clot formation. In this study, the authors investigated whether the hemostatic patterns had specific hemodynamic features using computational fluid dynamics (CFD) analysis. METHODS Twenty-six ruptured middle cerebral artery aneurysms were evaluated by 3D CT angiography and harvested at the time of clipping. The hemostatic patterns at the rupture points were assessed by means of histopathological examination, and morphological parameters were obtained. Transient analysis was performed, and wall shear stress-related hemodynamic parameters and invariant Q (vortex core region) were calculated. The morphological and hemodynamic parameters were compared among the hemostatic patterns. RESULTS Hematoxylin and eosin staining of the aneurysm wall showed 13 inside-pattern, 9 outside-pattern, and 4 other-pattern aneurysms. Three of the 26 aneurysms were excluded from further analysis, because their geometry models could not be generated due to low vascular CT values. Mann-Whitney U-tests showed that lower dome volume (0.04 cm3 vs 0.12 cm3, p = 0.014), gradient oscillatory number (0.0234 vs 0.0289, p = 0.023), invariant Q (-0.801 10-2/sec2 vs -0.124 10-2/sec2, p = 0.045) and higher aneurysm formation indicator (0.986 vs 0.963, p = 0.041) were significantly related to inside-pattern aneurysms when compared with outside-pattern aneurysms. CONCLUSIONS Inside-pattern aneurysms may have simpler flow patterns and less flow stagnation than outside-pattern aneurysms. CFD may be useful to characterize the hemostatic pattern of ruptured cerebral aneurysms.

Aneurysm Organization Effects of Gellan Sulfate Core Platinum Coil with Tenascin-C in a Simulated Clinical Setting and the Possible Mechanism

BACKGROUND This study aimed to deliver gellan sulfate core platinum coil with tenascin-C (GSCC-TNC) into rabbit side-wall aneurysms endovascularly and to evaluate the organization effects in a simulated clinical setting. METHODS Elastase-induced rabbit side-wall aneurysms were randomly coiled via a transfemoral route like clinical settings with platinum coils (PCs), gellan sulfate core platinum coils (GSCCs), or GSCC-TNCs (n = 5, respectively). Aneurysm-occlusion status was evaluated angiographically and histologically at 2 weeks post coiling. As each rabbit coiled aneurysm provided only 2-3 tissue slices due to technical limitations and prevented immunohistochemical evaluations, a PC, GSCC, or GSCC-TNC was randomly implanted in a rat blind-ended model (n = 3, respectively) and the organization effects were immunohistochemically evaluated for expressions of tenascin-C (TNC), transforming growth factor-beta (TGF-β), and matrix metalloproteinase-9 (MMP-9) 2 weeks later. RESULTS Coil handling was similar among the 3 kinds of coils. GSCCs showed a significantly higher ratio of organized area to the aneurysmal cavity than PCs, but GSCC-TNCs had the greatest organization-promoting effects on aneurysms (the ratio of organized area/aneurysmal luminal area: PC, 17.9 ± 7.1%; GSCC, 54.2 ± 18.3%; GSCC-TNC, 82.5 ± 5.8%). GSCC-TNCs had intense immunoreactivities for TNC, TGF-β, and MMP-9 in the organized thrombosis and tunica media. GSCCs also showed intense immunoreactivities for TNC, TGF-β, and MMP-9, although the extent was less than GSCC-TNCs. The immunoreactivities were hardly found in unorganized thrombus and the tunica media of aneurysm wall in the PC group. CONCLUSIONS This study first showed that GSCC-TNCs promote intra-aneurysmal clot organization in simulated clinical settings using rabbits possibly through the TGF-β and MMP-9 upregulation.

Hemodynamic characteristics of hyperplastic remodeling lesions in cerebral aneurysms

Background & purpose Hyperplastic remodeling (HR) lesions are sometimes found on cerebral aneurysm walls. Atherosclerosis is the results of HR, which may cause an adverse effect on surgical treatment for cerebral aneurysms. Previous studies have demonstrated that atherosclerotic changes had a correlation with certain hemodynamic characteristics. Therefore, we investigated local hemodynamic characteristics of HR lesions of cerebral aneurysms using computational fluid dynamics (CFD). Methods Twenty-four cerebral aneurysms were investigated using CFD and intraoperative video recordings. HR lesions and red walls were confirmed on the intraoperative images, and the qualification points were determined on the center of the HR lesions and the red walls. The qualification points were set on the virtual operative images for evaluation of wall shear stress (WSS), normalized WSS (NWSS), oscillatory shear index (OSI), relative residence time (RRT), and aneurysm formation indicator (AFI). These hemodynamic parameters at the qualification points were compared between HR lesions and red walls. Results HR lesions had lower NWSS, lower AFI, higher OSI and prolonged RRT compared with red walls. From analysis of the receiver-operating characteristic curve for hemodynamic parameters, OSI was the most optimal hemodynamic parameter to predict HR lesions (area under the curve, 0.745; 95% confidence interval, 0.603–0.887; cutoff value, 0.00917; sensitivity, 0.643; specificity, 0.893; P<0.01). With multivariate logistic regression analyses using stepwise method, NWSS was significantly associated with the HR lesions. Conclusions Although low NWSS was independently associated with HR lesions, OSI is the most valuable hemodynamic parameter to distinguish HR lesions from red walls.

A Case of Vertebral Artery Fusiform Aneurysm Treated by Flow Alteration: Successful Prediction of Therapeutic Effects Using Computational Fluid Dynamics

The treatment of intracranial complicated aneurysms remains challenging. In patients with complicated aneurysms that are neither clippable nor coilable, flow alteration treatment (FAT) with a combined procedure of proximal/distal occlusion or trapping of an aneurysm with bypass surgery has been reported. However, it is difficult to predict whatever FAT can achieve aneurysmal obliteration without ischemic complications. A 69-year-old female was incidentally diagnosed with a left vertebral artery (VA) fusiform aneurysm distal to the left posterior inferior cerebellar artery (PICA). Because one-year follow-up three-dimensional computed tomography angiography showed that the aneurysm grew significantly, surgical management was considered the therapy of choice. For determining treatment strategies, we assumed left VA occlusion at the proximal to the left PICA as a FAT model and performed computational fluid dynamics (CFD) analyses. The FAT model had much lower wall shear stress and shear rate at the aneurysm dome than presumed thresholds necessary to thrombus formation, while those at the PICA were obviously higher than the thresholds, and streamlines into the left PICA from the distal VA were preserved. These findings theoretically meant that surgical occlusion of the left VA proximal to the left PICA and aneurysm would induce intra-aneurysmal thrombus formation with preservation of the left PICA flow. The treatment was performed successfully and achieved the predicted results. CFD simulations may be useful to predict effects of FAT for complicated aneurysms.

P-018 Effect of pitavastatin on preventing ischaemic complications with carotid artery stenting: a multicentre prospective study

Background Carotid artery stenting (CAS) is becoming an alternative to carotid endarterectomy (CEA). However, periprocedural ischaemic stroke is one problem associated with CAS. Statin therapy can reportedly reduce periprocedural complication rates in coronary intervention. This study was designed to assess whether preoperative statin therapy reduces the risk of periprocedural ischaemic complications with CAS. Material and Methods In this prospective study at 11 centres, patients with carotid stenosis (symptomatic ≥50%, asymptomatic ≥80%) and at high risk of requiring CEA, but without previous statin treatment were divided into two groups by low-density lipoprotein cholesterol level (LDL-C). With LDL-C≥120 mg/dl, the previous stain (PS) group received pitavastatin at 4 mg/day for 4 weeks. With LDL-C <120 mg/dl, the non-PS group received no statin therapy for 4 weeks. After 4 weeks, both groups underwent CAS. Frequencies of new ipsilateral ischaemic lesions on diffusion-weighted imaging (DWI) within 72 h after CAS and major adverse events (MAEs) (defined as transient ischaemic attack, stroke, myocardial infarction or death) within 30 days were assessed. Results Among the 80 patients enrolled, 61 patients (PS group, n=31; non-PS group, n=30) fulfilled the inclusion criteria. New ipsilateral ischaemic lesions were identified in 8 of 31 patients (25.8%) in the PS group and 16 of 30 patients (55.3%) in the non-PS group (P=0.027). MAE occurred in 0 patients in the PS group and in 3 of 30 patients (10.0%) in the non-PS group (P=0.07). Conclusion Pretreatment with pitavastatin significantly reduced the frequency of periprocedural ischaemic complications associated with CAS. Disclosures K. Takayama: None. K. Myouchin: None. N. Ikeda: None. M. Sakamoto: None. I. Nakagawa: None. T. Wada: None. O. Masuo: None. I. Nakahara: None. H. Tanemura: None. N. Toma: None. M. Maeda: None. H. Suzuki: None. W. Taki: None.