ポスター(英語)5月18日(水）ポスター(英語)5月18日(水）

ポスター (英語)

5月18日(水）

18日

㈬ ポス ター

（英 語) Pe-001-1

Effect of pulsatility index on infarct volume in acute lacunar stroke

Department of Neurology, CHA Bundang Medical Center, CHA University, Seongnam, South Korea

○Jinkwon Kim，Yim Byeongsoo，Jonguk Kim，Kim Yoon，

Kim Ok Joon

Purpose: Lacunar stroke is a type of cerebral infarction caused by occlusion of a penetrating artery. Pulsatility index (PI) is an easily measurable transcranial Doppler ultrasound (TCD) parameter. PI reflects distal cerebral vascular resistance and has been interpreted as a surrogate marker of cerebral small vessel disease. We evaluated that an increased PI might be associated with infarct volume in acute lacunar stroke.Methods: We included 64 patients with acute lacunar stroke who underwent TCD and brain diffusion MRI. We investigated the association between mean PI value of bilateral middle cerebral arteries and infarct volume on diffusion weighted MRI using univariate and multivariate linear regression.Results: The mean infarct volume and PI were 482.18 ± 406.40 mm³ and 0.86 ± 0.18, respectively. On the univariate linear regression, there was a significant positive association between PI and infarct volume (p=0.001). In the multivariate model, a single standard deviation increase of PI (per 0.18) was significantly associated with an increase of 139.05 mm³in infarct volume (95％

confidence interval, 21.25 to 256.85; p=0.022).Conclusion: We demonstrated that PI was an independent determinant of infarct volume in acute lacunar stroke. The PI value measured in acute stroke may be a surrogate marker of the extent of ischemic injury.

Pe-001-2

Usefulness of WORSEN score: predictive score for the deterioration of acute ischemic stroke

1Department of Neurology, Juntendo Urayasu Hospital，²Department of Neurology, Juntendo University School of Medicine

○Nobukazu Miyamoto¹，Ryota Tanaka²，Yuji Ueno^1,2， Masao Watanabe¹，Yasutaka Tanaka²，Yoshiaki Shimada^1,2， Kazuo Yamashiro²，Nobutaka Hattori²，Takao Urabe¹

[Backgrouond] Early neurological worsening is associated with increased mortality and long-term functional disability. We developed WORSEN score for predicting factors of stroke deterioration within 1 week from onset, and investigated its usefulness.[Methods]

We retrospectively investigated 537 cerebral ischemia patients who were admitted to our hospital between April 2007 and March 2009. Deterioration of neurological findings was defined as worsening by 4 points or more of the NIHSS score from admission. Based on the results of our previous study, we developed the WORSEN score and used it to score the patients. Next, we applied the score to another 456 cerebral ischemia patients who were admitted to our hospitaland relational hospital between October 2013 and December 2014.

[Results] After multivariate analysis, we based the WORSEN score on the following factors:

wrong blood sugar control (W); old myocardial infarction (O); radiological findings (R);

size of infarct (S); elevated LDL-cholesterol (E); and neurological findings (N). Each factor was scored as 1 point. Next, we checked the WORSEN score in the first patient group.

Deterioration was noted in 35％ with a score of 3 points, 57％ with a score of 4, and 100％

with a score of 5. In the second patient group, deterioration was detected in 55.8％ with a score over 3 points.[Conclusion] Careful attention should be paid to acute stroke patients with high WORSEN scores. This scale might become an important tool for detecting neurological deterioration of ischemic stroke.

Pe-001-3

Infarct evolution in acute large artery occlusion of the anterior cerebral circulation

1Neurology Sevice, Tachikawa General Hospital，²Neurosurgery Sevice, Tachikawa General Hospital

○Hiroki Takano¹，Yohei Tanaka¹，Haruhiko Takahashi²， Yasushi Jimbo²，Hiroshi Abe²

Background: To clarify the time course of infarct evolution in acute large artery occlusion.

Methods: From our records, we selected consecutive patients with (1) ischemic stroke with known onset time, (2) MRA (magnetic resonance arteriography) and DWI (diffusion-weighted imaging) performed within 300 minutes after onset, and (3) MRA- documented ipsilateral occlusions at M1 of middle cerebral artery (MCA), or internal carotid artery (ICA) and MCA. Infarcts were scored by ASPECTS+W (the modified ASPECTS for DWI use). Results: Sixty-five patients were identified including 47 with M1 occlusion (mean age; 76 years) and 18 with ICA-M1 occlusion (mean age; 78 years). The mean time from onset to imaging in the M1 occlusion group was 131.7 minutes which was longer than that in the ICA-M1 occlusion group of 92.9 minutes (P=0.012); however, the median ASPECTS+W in the M1 group was 8 which was higher than that in the ICA-M1 group of 2 (P=0.0003). The proportion of patients with ASPECTS+W＞7 was 70％ in the M1 group, and 33％ in the ICA-M1 group (P=0.007). Even though only the patients with time to imaging of less than 150 minutes were analyzed, the proportion with ASPECTS+

W＞7 was a little increased to 81％ in the M1 group (P=0.1), and that was not so changed at 35％ in the ICA-M1 group (P=0.9). In both groups, ASPECTS+W did not correlate with time after onset. Conclusions: The acute phase infarct evolution in the proximal anterior circulation occlusion was not time-dependent. Apparently, occlusion site was more important. The current time-based recanalization therapy might be reconsidered.

Pe-001-4

The SVS with two compositions and single cortical infarct on DWI are specific to cardioembolism

1Department of Clinical neurosciences, Tokushima University，²Department of Neurosurgery, Tokushima University

○Nobuaki Yamamoto¹，Yuki Yamamoto¹，Yuki Unai¹，Yuishin Izumi¹， Ryuji Kaji¹，Junichiro Satomi²，Shinji Nagahiro²

Background:Although accurate diagnosis of the ischemic stroke subtype is one of the most important factors for selection of therapeutic approach, it is sometimes difficult at the time of admission. We studied independent predictive markers of magnetic resonance imaging (MRI), including two-layered SVS for diagnosing cardioembolism.Methods:We included 132 ischemic stroke patients within 24 hours from onset who suffered internal carotid artery or middle cerebral artery occlusion due to cardioembolism (group CE) or large artery atherosclerosis (group LAA) . We studied about independent markers on MRI such as two- layered SVS and abnormal finding patterns of diffusion-weighted image (DWI) for predicting cardioembolism.Results:In this study, 132 patients (72 men and 60 women, age 74.5 ± 12.1 years) were included. Of these, 63 (47.7 ％) were cardioembolism. In univariate analysis, frequency of comorbid atrial fibrillation, presence of two-layered SVS on T2*-WI and that of single corticosubcortical infarct on DWI were significantly higher in group CE. In multivariate analysis, the presence of two-layered SVS and single corticosubcortical infarct were the independent markers for diagnosing cardioembolism. (odds ratio, two-layered SVS, 29.41, p ＜ 0.001; single corticosubcortical infarct, 10.80, p = 0.043) . Conclusion: Independent markers for predicting cardioembolism may be two- layered SVS on T2*-WI and single corticosubcortical infarct on DWI.

Pe-001-5

Malignant DWI Profile assessment in Acute Ischemic Stroke

1Showa University Fujigaoka Hospital, Department of Neurology，²Showa University Fujigaoka Hospital Department of Neurosurgery

○Manabu Inoue¹，Joe Matsuzaki²，Nomoto Shohei¹，Natsuko Iizuka¹， Hiroaki Iwanami¹，Yuki Shimizu¹，Kazuhiro Itaya¹，Hiroo Ichikawa¹ [Objective] The efficacy of reperfusion therapy including endovascular therapy (EVT) in acute stroke is established but the imaging inclusion/exclusion criterion has not been assessed. The "Malignant profile" is a magnetic resonance imaging (MRI) pattern that is associated with poor outcomes. The aim of this study is to estimate the Malignant profile identified by diffusion weighted image (DWI) in patients treated with reperfusion therapy.[Methods]Acute anterior ischemic stroke patient obtained DWI at baseline before reperfusion therapy was included. Outcome was assessed by modified Rankin Scale (mRS) and DWI volume was measured by automated software.

ROC curve analysis was performed to identify core volume thresholds in patients with poor outcome (mRS 4-6).[Results]Twenty five patients achieved reperfusion therapy and 16 had interpretable DWI scan. Mean age was 79±10 years and median NIHSS was 22 (14-29). Time to imaging (median, IQR) was 104 (66-104) minutes, DWI volume was 57.3 (16.2-80.1) mL, and time to reperfusion therapy was 190 (138-285) minutes. ROC analysis determined the best DWI volume measurement for Malignant profile as 49.1 mL (87.5％ specificity and 100％ sensitivity, p＜0.01). Out of 16 patients, 14 underwent to EVT and 10 had recanalization (71％). Of these, 50％ (5/10) had poor outcome and median DWI volume (IQR) was 69.3 (66.9-99.7) mL. [Conclusions]

Malignant profile on DWI is approximately 49 mL in reperfusion therapy-eligible patients. The clinical outcome of these patients is poor despite recanalization and the indication should be well considered.

Pe-002-1

Cerebral small vessel diseases and mild parkinsonian signs in the elderly with vascular risk factors

1Department of Neurology, Osaka University Graduate School of Medicine，²Department of Neurology and Stroke Center, Kinki University Graduate School of Medicine，³Department of Stroke Medicine, Kawasaki Medical University Graduate School of Medicine，⁴Department of Neurology, Tokyo Women’s Medical University Graduate School of Medicine

○Jun Hatate¹，Kaori Miwa¹，Mari Matsumoto²，Tsutomu Sasaki¹，Yoshiki Yagita³， Manabu Sakaguchi¹，Kazuo Kitagawa⁴，Hideki Mochizuki¹

[Objective] The aim of this study was to examine the association between mild parkinsonian signs (MPS) and cerebral small-vessel disease (SVD) and total SVD burden in patients with vascular risk factors. [Methods] We performed a cross-sectional study among 268 outpatients without parkinsonism or dementia (71.0±7.8 years, 63％ male). MPS was evaluated via Unified Parkinson’s Disease Rating Scale PartIII. Brain MRI was used to determine SVD (cerebral microbleeds [CMBs], lacunar infarctions [LIs], and white matter hyperintensities [WMH]) . We rated the total SVD score, by counting the presence of each of SVD feature (LIs, CMBs, periventricular hyperintensities [PVH], and deep WMH [DWMH]). Logistic regression analyses were performed adjusting for age, sex, history of stroke, hypertension, diabetes mellitus, and dyslipidemia. [Results] In a multivariate analysis, we found that the presence of deep CMBs, mixed (in the basal ganglia and thalamus) LIs, PVH, DWMH, and total SVD score were significantly associated with MPS, whereas strictly lobar CMBs and other LIs (in strictly basal ganglia or thalamus) were not. We found an association between mixed LIs, PVH, DWMH, and total SVD and gait/balance function, between PVH and rigidity, and between mixed LIs and bradykinesia. Among elderly participants (≧73 years), the association of total SVD, deep CMBs, mixed LIs, and PVH, with MPS remained significant. [Conclusions] SVD including CMBs, especially total SVD score, might be a surrogate marker for MPS and support the contribution of hypertensive microangiopathy as the underlying etiology.

- 479 -

18日

㈬ ポス ター

（英 語) Pe-001-1

Effect of pulsatility index on infarct volume in acute lacunar stroke

Department of Neurology, CHA Bundang Medical Center, CHA University, Seongnam, South Korea

○Jinkwon Kim，Yim Byeongsoo，Jonguk Kim，Kim Yoon，

Kim Ok Joon

Purpose: Lacunar stroke is a type of cerebral infarction caused by occlusion of a penetrating artery. Pulsatility index (PI) is an easily measurable transcranial Doppler ultrasound (TCD) parameter. PI reflects distal cerebral vascular resistance and has been interpreted as a surrogate marker of cerebral small vessel disease. We evaluated that an increased PI might be associated with infarct volume in acute lacunar stroke.Methods: We included 64 patients with acute lacunar stroke who underwent TCD and brain diffusion MRI. We investigated the association between mean PI value of bilateral middle cerebral arteries and infarct volume on diffusion weighted MRI using univariate and multivariate linear regression.Results: The mean infarct volume and PI were 482.18 ± 406.40 mm³ and 0.86 ± 0.18, respectively. On the univariate linear regression, there was a significant positive association between PI and infarct volume (p=0.001). In the multivariate model, a single standard deviation increase of PI (per 0.18) was significantly associated with an increase of 139.05 mm³in infarct volume (95％

confidence interval, 21.25 to 256.85; p=0.022).Conclusion: We demonstrated that PI was an independent determinant of infarct volume in acute lacunar stroke. The PI value measured in acute stroke may be a surrogate marker of the extent of ischemic injury.

Pe-001-2

Usefulness of WORSEN score: predictive score for the deterioration of acute ischemic stroke

1Department of Neurology, Juntendo Urayasu Hospital，²Department of Neurology, Juntendo University School of Medicine

○Nobukazu Miyamoto¹，Ryota Tanaka²，Yuji Ueno^1,2， Masao Watanabe¹，Yasutaka Tanaka²，Yoshiaki Shimada^1,2， Kazuo Yamashiro²，Nobutaka Hattori²，Takao Urabe¹

[Backgrouond] Early neurological worsening is associated with increased mortality and long-term functional disability. We developed WORSEN score for predicting factors of stroke deterioration within 1 week from onset, and investigated its usefulness.[Methods]

We retrospectively investigated 537 cerebral ischemia patients who were admitted to our hospital between April 2007 and March 2009. Deterioration of neurological findings was defined as worsening by 4 points or more of the NIHSS score from admission. Based on the results of our previous study, we developed the WORSEN score and used it to score the patients. Next, we applied the score to another 456 cerebral ischemia patients who were admitted to our hospitaland relational hospital between October 2013 and December 2014.

[Results] After multivariate analysis, we based the WORSEN score on the following factors:

wrong blood sugar control (W); old myocardial infarction (O); radiological findings (R);

size of infarct (S); elevated LDL-cholesterol (E); and neurological findings (N). Each factor was scored as 1 point. Next, we checked the WORSEN score in the first patient group.

Deterioration was noted in 35％ with a score of 3 points, 57％ with a score of 4, and 100％

with a score of 5. In the second patient group, deterioration was detected in 55.8％ with a score over 3 points.[Conclusion] Careful attention should be paid to acute stroke patients with high WORSEN scores. This scale might become an important tool for detecting neurological deterioration of ischemic stroke.

Pe-001-3

Infarct evolution in acute large artery occlusion of the anterior cerebral circulation

1Neurology Sevice, Tachikawa General Hospital，²Neurosurgery Sevice, Tachikawa General Hospital

○Hiroki Takano¹，Yohei Tanaka¹，Haruhiko Takahashi²， Yasushi Jimbo²，Hiroshi Abe²

Background: To clarify the time course of infarct evolution in acute large artery occlusion.

Methods: From our records, we selected consecutive patients with (1) ischemic stroke with known onset time, (2) MRA (magnetic resonance arteriography) and DWI (diffusion-weighted imaging) performed within 300 minutes after onset, and (3) MRA- documented ipsilateral occlusions at M1 of middle cerebral artery (MCA), or internal carotid artery (ICA) and MCA. Infarcts were scored by ASPECTS+W (the modified ASPECTS for DWI use). Results: Sixty-five patients were identified including 47 with M1 occlusion (mean age; 76 years) and 18 with ICA-M1 occlusion (mean age; 78 years). The mean time from onset to imaging in the M1 occlusion group was 131.7 minutes which was longer than that in the ICA-M1 occlusion group of 92.9 minutes (P=0.012); however, the median ASPECTS+W in the M1 group was 8 which was higher than that in the ICA-M1 group of 2 (P=0.0003). The proportion of patients with ASPECTS+W＞7 was 70％ in the M1 group, and 33％ in the ICA-M1 group (P=0.007). Even though only the patients with time to imaging of less than 150 minutes were analyzed, the proportion with ASPECTS+

W＞7 was a little increased to 81％ in the M1 group (P=0.1), and that was not so changed at 35％ in the ICA-M1 group (P=0.9). In both groups, ASPECTS+W did not correlate with time after onset. Conclusions: The acute phase infarct evolution in the proximal anterior circulation occlusion was not time-dependent. Apparently, occlusion site was more important. The current time-based recanalization therapy might be reconsidered.

Pe-001-4

The SVS with two compositions and single cortical infarct on DWI are specific to cardioembolism

1Department of Clinical neurosciences, Tokushima University，²Department of Neurosurgery, Tokushima University

○Nobuaki Yamamoto¹，Yuki Yamamoto¹，Yuki Unai¹，Yuishin Izumi¹， Ryuji Kaji¹，Junichiro Satomi²，Shinji Nagahiro²

Background:Although accurate diagnosis of the ischemic stroke subtype is one of the most important factors for selection of therapeutic approach, it is sometimes difficult at the time of admission. We studied independent predictive markers of magnetic resonance imaging (MRI), including two-layered SVS for diagnosing cardioembolism.Methods:We included 132 ischemic stroke patients within 24 hours from onset who suffered internal carotid artery or middle cerebral artery occlusion due to cardioembolism (group CE) or large artery atherosclerosis (group LAA) . We studied about independent markers on MRI such as two- layered SVS and abnormal finding patterns of diffusion-weighted image (DWI) for predicting cardioembolism.Results:In this study, 132 patients (72 men and 60 women, age 74.5 ± 12.1 years) were included. Of these, 63 (47.7 ％) were cardioembolism. In univariate analysis, frequency of comorbid atrial fibrillation, presence of two-layered SVS on T2*-WI and that of single corticosubcortical infarct on DWI were significantly higher in group CE. In multivariate analysis, the presence of two-layered SVS and single corticosubcortical infarct were the independent markers for diagnosing cardioembolism. (odds ratio, two-layered SVS, 29.41, p ＜ 0.001; single corticosubcortical infarct, 10.80, p = 0.043) . Conclusion: Independent markers for predicting cardioembolism may be two- layered SVS on T2*-WI and single corticosubcortical infarct on DWI.

Pe-001-5

Malignant DWI Profile assessment in Acute Ischemic Stroke

1Showa University Fujigaoka Hospital, Department of Neurology，²Showa University Fujigaoka Hospital Department of Neurosurgery

○Manabu Inoue¹，Joe Matsuzaki²，Nomoto Shohei¹，Natsuko Iizuka¹， Hiroaki Iwanami¹，Yuki Shimizu¹，Kazuhiro Itaya¹，Hiroo Ichikawa¹ [Objective] The efficacy of reperfusion therapy including endovascular therapy (EVT) in acute stroke is established but the imaging inclusion/exclusion criterion has not been assessed. The "Malignant profile" is a magnetic resonance imaging (MRI) pattern that is associated with poor outcomes. The aim of this study is to estimate the Malignant profile identified by diffusion weighted image (DWI) in patients treated with reperfusion therapy.[Methods]Acute anterior ischemic stroke patient obtained DWI at baseline before reperfusion therapy was included. Outcome was assessed by modified Rankin Scale (mRS) and DWI volume was measured by automated software.

ROC curve analysis was performed to identify core volume thresholds in patients with poor outcome (mRS 4-6).[Results]Twenty five patients achieved reperfusion therapy and 16 had interpretable DWI scan. Mean age was 79±10 years and median NIHSS was 22 (14-29). Time to imaging (median, IQR) was 104 (66-104) minutes, DWI volume was 57.3 (16.2-80.1) mL, and time to reperfusion therapy was 190 (138-285) minutes. ROC analysis determined the best DWI volume measurement for Malignant profile as 49.1 mL (87.5％ specificity and 100％ sensitivity, p＜0.01). Out of 16 patients, 14 underwent to EVT and 10 had recanalization (71％). Of these, 50％ (5/10) had poor outcome and median DWI volume (IQR) was 69.3 (66.9-99.7) mL. [Conclusions]

Malignant profile on DWI is approximately 49 mL in reperfusion therapy-eligible patients. The clinical outcome of these patients is poor despite recanalization and the indication should be well considered.

Pe-002-1

Cerebral small vessel diseases and mild parkinsonian signs in the elderly with vascular risk factors

1Department of Neurology, Osaka University Graduate School of Medicine，²Department of Neurology and Stroke Center, Kinki University Graduate School of Medicine，³Department of Stroke Medicine, Kawasaki Medical University Graduate School of Medicine，⁴Department of Neurology, Tokyo Women’s Medical University Graduate School of Medicine

○Jun Hatate¹，Kaori Miwa¹，Mari Matsumoto²，Tsutomu Sasaki¹，Yoshiki Yagita³， Manabu Sakaguchi¹，Kazuo Kitagawa⁴，Hideki Mochizuki¹

[Objective] The aim of this study was to examine the association between mild parkinsonian signs (MPS) and cerebral small-vessel disease (SVD) and total SVD burden in patients with vascular risk factors. [Methods] We performed a cross-sectional study among 268 outpatients without parkinsonism or dementia (71.0±7.8 years, 63％ male). MPS was evaluated via Unified Parkinson’s Disease Rating Scale PartIII. Brain MRI was used to determine SVD (cerebral microbleeds [CMBs], lacunar infarctions [LIs], and white matter hyperintensities [WMH]) . We rated the total SVD score, by counting the presence of each of SVD feature (LIs, CMBs, periventricular hyperintensities [PVH], and deep WMH [DWMH]). Logistic regression analyses were performed adjusting for age, sex, history of stroke, hypertension, diabetes mellitus, and dyslipidemia. [Results] In a multivariate analysis, we found that the presence of deep CMBs, mixed (in the basal ganglia and thalamus) LIs, PVH, DWMH, and total SVD score were significantly associated with MPS, whereas strictly lobar CMBs and other LIs (in strictly basal ganglia or thalamus) were not. We found an association between mixed LIs, PVH, DWMH, and total SVD and gait/balance function, between PVH and rigidity, and between mixed LIs and bradykinesia. Among elderly participants (≧73 years), the association of total SVD, deep CMBs, mixed LIs, and PVH, with MPS remained significant. [Conclusions] SVD including CMBs, especially total SVD score, might be a surrogate marker for MPS and support the contribution of hypertensive microangiopathy as the underlying etiology.

- 480 -

18日

㈬ ポス ター

（英 語)

Pe-002-2

Natural history of cerebral microbleeds in a prospective study

Internal medicine cardiovascular, respiratory and neurology division Asahikawa Medical University

○Tsukasa Saito，Nobuyuki Sato，Yuichiro Kawamura，

Naoyuki Hasebe

[Purpose] Cerebral microbleeds (CMBs) are widely accepted as a marker of vulnerability of the cerebral small vessels. However, the natural history of CMBs remains largely unknown. This study aimed to clarify the natural history of CMBs in a prospective manner.[Method] We performed yearly brain magnetic resonance imaging (MRI) assessments for 5 or more years in 36 non-valvular atrial fibrillation outpatients.[Results] Among the 36 patients, 8 had CMBs present at baseline and 4 showed new CMBs on follow-up. We followed up 8 patients for 7 years, 2 patients for 8 years and 2 patients for 6 years. The CMBs disappeared in 4 patients during the follow-up duration. In 5 patients, the CMBs remained no- change over 7 years. Importantly, we observed new appearance of CMBs in 2 patients, and they disappeared after 2 and 4 years from their appearance, respectively.[Conclusion] We could find the natural history of CMBs using yearly performed MRIs for over 5 years in a prospective manner. Most CMBs seemed to remain for over 7 years. Because of its refined nature, the detection and confirmation of CMBs are not always perfectly impeccable. Further surveillance in many subjects and a long-term prospective follow-up are required.

Pe-002-3

The Distribution of Cerebral Microbleeds Determines Their Association with Vascular Resistance

Department of Neurology, Ewha Womans University School of Medicine, Seoul, Korea

○Yoonkyung Chang，Gyeongseon Choi，A-reum Jung，Minjung Youn，

Yong-Jae Kim，Tae-Jin Song

Objective: Cerebral microbleeds (CMBs), are assumed to be manifestations of cerebral small vessel disease. Meanwhile, the pulsatility index (PI) measured by transcranial Dopplerdoppler, has long been proposed to reflect vascular resistance.

Therefore, we investigated whether the presence and location of CMBs are associated with PI.Methods: Between January 2007 and December 2012, we enrolled 702 consecutive patients diagnosed with noncardioembolic acute ischemic stroke mechanism. The peak systolic velocity, end diastolic velocity and PI were investigated in bilateral middle cerebral arteries (MCA) and mean values of PI acquired from bilateral/and or unilateral MCAs were entered for analysis. Binary and multinomial logistic regression analysis was used for determining factors related to the existence of CMBs according to their location.Results: The mean age was 62 ±11 years and 69.1％ were male. PI was higher in patients with CMBs (1.02 ± 0.25) than in those without (0.86 ± 0.36) (p=0.002). Moreover, PI was higher in the nonlobar CMBs group than the strictly lobar CMBs group (p=0.016). In multivariate multinomial logistic regression, PI was independently associated with the nonlobar CMBs group (odds ratio: 2.99, 95％ confidence interval: 1.12 - 7.97, p = 0.042), but not with the strictly lobar CMBs group.Conclusions: PI, which representing cerebral vascular resistance, was independently associated with nonlobar CMBs, but not strictly lobar CMBs. These findings suggest a pathophysiologic association between PI and CMBs in the nonlobar region.

Pe-002-4 海外最優秀候補演題

Characterization of CADASIL among the Han Chinese in Taiwan: Distinct Genotypic and Phenotypes

1Department of Neurology, Taipei Veterans General Hospital, Taipei, Taiwan，

2Department of Neurology, National Yang-Ming University School of Medicine, Taipei, Taiwan，³Brain Research Center, National Yang-Ming University, Taipei, Taiwan

○Yi-chu Liao^1,2，Cheng-Tsung Hsiao^1,2，Jong-Ling Fuh^1,2,3，Yo-Tsen Liu^1,2， Bing-Wen Soong^1,2,3，Yi-Chung Lee^1,2,3

Objective Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is originally featured with a strong clustering of mutations in NOTCH3 exons 3-6 and leukoencephalopathy with frequent anterior temporal pole involvement.MethodsMutation analyses of exons 2 to 24 of NOTCH3 were performed by Sanger sequencing. Haplotype analysis was done by genotyping 6 polymorphic microsatellite markers flanking NOTCH3 and covering a region of 7.54 kcM.ResultsA total of 112 CADASIL patients from 95 families were included. Twenty different mutations in NOTCH3 were uncovered, including 3 novel ones, and R544C in exon 11 was the most common mutation, accounting for 70.5％ of the pedigrees. Haplotype analyses were conducted in 14 families harboring NOTCH3 R544C mutation and demonstrated a common haplotype linked to NOTCH3 R544C at loci D19S929 and D19S411. Comparing with CADASIL in most Caucasian populations, CADASIL in Taiwan has several distinct features, including less frequent anterior temporal involvement, older age at symptom onset, higher incidence of intracerebral hemorrhage, and rarer occurrence of migraine. Subgroup analyses revealed that the R544C mutation is associated with lower frequency of anterior temporal involvement, later age at onset and higher frequency of cognitive dysfunction.ConclusionsThe present study broadens the spectrum of NOTCH3 mutations and provides additional insights for the clinical and molecular characteristics of CADASIL patients of Han-Chinese descents.

Pe-002-5

Diagnostic value of skin biopsy immunostaining with a Notch3 antibody for CADASIL

1Department of Neurology, Graduate School of Medical Sciences, Kumamoto University，²Department of Stroke and Cerebrovascular Medicine, Kyorin University Faculty of Medicine

○Akihiko Ueda¹，Mitsuharu Ueda¹，Akihito Nagatoshi¹， Makoto Nakajima¹，Teruyuki Hirano²，Yukio Ando¹

Background and purpose: Detection of granular osmiophilic material (GOM) by electron microscopy has been established as pathologic diagnosis for cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) . Immunohistohemical analyses for abnormal accumulation of Notch3 ectodomain are also useful for pathologic diagnosis methods. However, these methods have not been established as worldwide diagnostic methods. In this study, using frozen sections of skin samples, we performed immunohistochemical analyses of Notch3 and compared utility of immunohistochemical analyses of Notch3 with that of GOM detection by electron microscopy.Material and Methods: Skin biopsy specimens from upper arm were obtained in 21 suspected CADASIL patients from 2007 to 2015. We performed immunohistochemical stains of Notch3 with Victoria Blue stains to detect internal elastic lamina of arterioles.

Definite diagnosis of CADASIL was defined as detection of Notch3 mutation by genetic tests or GOM detection by electron microscopy.Results: All the sections in the first biopsy were available for detection of representative arteries. Seventeen patients had abnormal accumulations of Notch3. All of them were diagnosed as CADASIL. Four patients did not have abnormal accumulation of Notch3. All of them did not have any GOM deposits by electron microscopy.Conclusion: Immunohistochemical analyses of Notch3 in frozen skin sections are useful for screening of diagnosis of CADASIL.

Pe-002-6

In vivo detection of cerebral cortical microinfarts with 3 Tesla MRI in CADASIL Peking University First Hospital

○Xiaojing Fang，Zhaoxia Wang，yun Yuan，Yu Lei，jiangxi Xiao Objective:CADASIL is an inherited small-vessel disease caused by mutations in NOTCH3. MRI reveals leucoaraiosis with multiple lacunar infarcts in the deep white matter. Recent studies suggested that the cerebral cortex may be involved in small vessel disease (SVD) of the brain, while the data on CADASIL is still limited. While Cortical microinfarcts (CMIs) are best detected by 7T postmortem MRI, some studies proved that CMI is also retraceable on 3T MRI. Here we try to identify cortical microinfarcts on 3T MRI in CADASIL patients.Methods:Nine patients with CADASIL was enrolled in this study, whose diagnosis were confirmed by NOTCH3 gene mutation testing, and skin biopsy finding of GOM on the surface of smooth muscle cells of the arterioles. The age was between 42 and 59 years old. All of these patients presented with recurrent stroke with cognitive decline over 4 to 15 years. 3.0T MRI , 3-dimensional T1-weighted, 3D fluid- attenuated inversion recovery were performed in all patients.Results:3.0T MRI showed that all patients presented cerebral microinfarcts. The number of CMIs varied from 3 to 21 in different patients. CMIs located mostly in the deep layer of cortex with involving the subcortical white matter. The molecular layer was involved in none of them.Conclusions:This study provides strong evidence that cortical microinfarcts is common in CADASIL, which can be detected with 3.0T MRI.

Pe-002-7

Distinct molecular mechanisms of HTRA1 mutants in manifesting heterozygotes with CARASIL

1Department of Neurology, Brain Research Institute, Niigata University，²Department of Molecular Neuroscience, Brain Research Institute, Niigata University，³Department of Neurology, Kyoto Prefectural University of Medicine，⁴Department of Neurology, Ichinomiya Municipal Hospital，⁵Department of Neurology, Nishi-Niigata Chuo National Hospital，⁶Department of Neurology, Shiseikai-Daini Hospital，⁷Department of Neurology, Kanazawa Medical University，⁸Department of Neurology, Chiba University，⁹Department of Neurology, Nantan General Hospital，¹⁰Institute for Medical Science of Aging, Aichi Medical University

○Hiroaki Nozaki¹，Taisuke Kato²，Megumi Nihonmatsu¹，Yohei Saito¹，Ikuko Mizuta³，Tomoko Noda⁴， Ryoko Koike⁵，Kazuhide Miyazaki⁶，Muichi Kaito⁷，Shoichi Ito⁸，Masahiro Makino⁹，Akihide Koyama²， Atsushi Shiga²，Masahiro Uemura¹，Yumi Sekine¹，Kenju Hara¹，Mari Yoshida¹⁰，Masatoyo Nishizawa¹， Toshiki Mizuno³，Osamu Onodera²

[Objective]To investigate how mutant HTRA1s contribute to cerebral small vessel disease in a heterozygote state.[Methods]We recruited 113 unrelated index patients younger than 70 years of age with severe white matter hyperintensities. The coding sequences in the HTRA1 gene were analyzed. We evaluated HTRA1 protease activities using FITC-labeled casein and oligomeric HTRA1 formation using gel filtration chromatography.[Results]We found four heterozygous missense mutations in the HTRA1 gene (p.G283E, p.P285L, p.R302Q, and p.T319I) in six patients from 113 unrelated index patients and two siblings in two unrelated families with p.R302Q. The mean age at onset for cognitive impairment was 51.1 years. An autopsied case with p.G283E showed arteriopathy in cerebral small arteries. These mutant HTRA1s showed markedly decreased protease activities and inhibited wild-type HTRA1 activity in a mixture condition, whereas two of three mutant HTRA1s reported in CARASIL (A252T and V297M) did not inhibit wild-type HTRA1 activity.

The P285L and R302Q HTRA1, observed in manifesting heterozygotes, formed trimers and have mutations in the domains that are important for trimer-associated HTRA1 activation. In contrast, A252T and V297M HTRA1, observed in CARASIL, also formed trimers but have mutations outside the domains. The other mutant HTRA1s observed in manifesting heterozygotes (G283E and T319I) failed to properly fold.[Conclusion]The mutant HTRA1s observed in manifesting heterozygotes have distinct molecular features compared to mutant HTRA1s observed in CARASIL.

18日

㈬ ポス ター

（英 語) Pe-003-1

TNF-α is associated with the mechanism for leptomeningeal arteriogenesis impairment in db/db mice

1Department of Neurology, Osaka University Graduate School of Medicine，²Department of Stroke Medicine, Kawasaki Medical School，³Department of Neurology, Tokyo Women’s Medical University

○Toshiro Yukami¹，Yoshiki Yagita²，Hideaki Kanki¹，Akihiro Watanabe¹，Naoki Oyama¹， Yasukazu Terasaki¹，Tsutomu Sasaki¹，Manabu Sakaguchi¹，Kazuo Kitagawa³， Hideki Mochizuki¹

[Objective] Leptomeningeal arteriogenesis is a key factor that defines the severity of ischemic stroke. Patients with stroke generally have vascular risk factors, such as diabetes. We found that leptomeningeal arteriogenesis 14 days after unilateral common carotid artery occlusion was impaired in db/db mice. This study was performed to examine the mechanism for leptomeningeal arteriogenesis impairment in db/db mice. [Methods] Macrophages play an important role in the process of arteriogenesis. We performed immunostaining using Mac-2 antibody to assess the degree of macrophage accumulation on the dorsal surface of the brain 7 days after CCA occlusion in db/+ and db/db mice. Next, we investigated the mRNA expression of several macrophage-related factors in the left cerebral cortex, including leptomeningeal anastomoses, in db/+ and db/db mice using real-time polymerase chain reaction. Finally, we tested whether the leptomeningeal arteriogenesis could be restored by pharmaceutical intervention in the db/db mice. [Results] The number of Mac-2-positive cells was increased and TNF-α mRNA expression was induced after common carotid artery occlusion in the db/+ mice.

However, these responses were not observed in the db/db mice. Administration of etanercept (TNF-α inhibitor) before common carotid artery occlusion restored the hypoperfusion-induced leptomeningeal arteriogenesis in db/db mice. [Conclusions] These results indicate that suppression of the TNF-α response to hypoperfusion is the major contributing factor for the leptomeningeal arteriogenesis impairment in db/db mice.

Pe-003-2

Heteroduplex oligonucleotide reduced gene expression in focal ischemic brain in mice

1Department of Neurology and Neurological Science, Tokyo Medical and Dental University，²Institute Multidisciplinary Research for Advanced Materials, Tohoku University

○Fuying Li¹，Satoru Ishibashi¹，Kotaro Yoshioka¹，Takehiko Wada²， Masahiko Ichijo¹，Eri Iwasawa¹，Jindong Song¹，Yongquan Zhang¹， Takanori Yokota¹

[Backgrounds]Tocopherol-conjugated heteroduplex oligonucleotide (Toc-HDO) exerts a strong gene silencing effect on many organs, but the delivery of Toc-HDO to the brain is very limited by the blood brain barrier (BBB). Since BBB is disrupted at the acute phase of stroke, intravenously administered (i.v.) Toc-HDO may further penetrate the BBB and reduce target gene expression primarily in ischemic region.[Methods]The permanent middle cerebral artery occlusion (pMCAO) was induced in male C57BL/6 mice (n=60). Toc-HDO targeting mMALAT1 was administrated via tail vein at 24 hours after pMCAO, and the MALAT1 gene expression of brain was quantified by RT-PCR. The distribution of fluorescence labeled Toc-HDO in the ischemic brain was investigated under confocal microscopy with cell specific markers.[Results]Three days after Toc-HDO administration, MALAT1 mRNA levels decreased in the ischemic region at 24 hours after pMCAO; the mMALAT1 levels in pMCAO+Toc-HDO group was significantly reduced to 30.1±9.5％ compared with pMCAO+PBS controls (p ＜ 0.01). Fluorescence labeled Toc-HDO distributed in the ischemic brain, primarily detected in neuronal and endothelial cells, but not in astrocytes. Total infarction volume was significantly larger in pMCAO+Toc-HDO group compared with controls.[Conclusion]We found a successful gene silencing effect of the ischemic brain by means of i.v. administration of Toc-HDO targeting mMALAT1, which exacerbates ischemic injury. Our findings establish proof of concept for the application of HDO to gene silencing therapy for acute stroke.

Pe-003-3

Serine racemase inhibition induces NO-mediated neurovascular protection during cerebral ischemia

Department of Neurology, Osaka University Graduate School of Medicine

○Akihiro Watanabe，Tsutomu Sasaki，Toshiro Yukami，Hideaki Kanki，

Manabu Sakaguchi，Hideki Mochizuki

[Purpose] There are no effective neuroprotectant drugs for acute cerebral ischemia. Serine racemase (SR) synthesizes D-serine, which is involved in N- methyl-D-aspartate (NMDA) receptor-induced neurotoxicity. However, regulatory mechanisms controlling SR-activity in the neurovascular unit during cerebral ischemia remain to be clarified. So, we investigated the effects of SR inhibition on neurovascular protection after ischemia.[Methods] We investigated whether the SR inhibitor phenazine methosulfate (PMS) could alleviate neuronal damage in an in vitro ischemic model (oxygen glucose deprivation [OGD]) using neuron cultures and in an in vivo mouse model of ischemia (middle cerebral artery occlusion [MCAO]) . We examined the effect of PMS on SR expression and phosphorylation by western blotting. Changes in cerebral blood flow (CBF) associated with administration of PMS were detected by laser speckle imaging.

Finally, we tested whether NO production was affected by PMS-administration after ischemia both in vitro and in vivo.[Results] PMS decreased cell death after OGD in neuron cultures. Infarct volumes were also reduced in PMS-treated mice.

PMS inhibited SR phosphorylation after ischemia. Reductions in regional CBF after MCA occlusion were improved by administration of PMS. Treatment with PMS increased both phosphorylation of eNOS and NO production.[Conclusions]

These findings indicate that SR inhibition acts as a neuroprotectant in the neurovascular unit and ameliorant of CBF abnormalities post-stroke. Thus, pharmacologic SR inhibition has potential clinical applications.

Pe-003-4

Telmisartan Treatment Strongly Improved ApoE/LDL-R Signals in Post-Stroke SHR-SR

Department of Neurology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences

○Toru Yamashita，Yun Zhai，Kota Sato，Mami Takemoto，

Nozomi Hishikawa，Yasuyuki Ohta，Koji Abe

Background: Telmisartan, an angiotensin receptor blocker also called metabosartan,is a promising solution for preventing cognitive decline or the incidence of dementia.Methods:We examined the effects of telmisartan on cholesterol transport- related proteins (apolipoprotein E (ApoE)/low-density lipoprotein receptor (LDL- R) ) and microtubule-associated protein 2 (MAP2) in the brain of spontaneously hypertensive stroke resistant (SHR-SR). SHR-SR received transient middle cerebral artery occlusion (tMCAO) for 90 minutes at 12 weeks of age and then was divided into 3 experiment groups including a vehicle, low-dose telmisartan (0.3 mg/kg/day), and high-dose telmisartan (3 mg/kg/day).Results: The low dose served to improve the metabolic syndrome of SHR-SR without lowering the blood pressure (BP) whereas the high dose was used to improve metabolic syndrome while lowering BP.

Immunohistologic analysis showed that ApoE expression of cortical neurons was strong in the vehicle group at 6, 12, and 18 months of age. On the other hand, LDL-R expression was transiently increased at 6 months of age only on the ipsilateral side.

Telmisartan dramatically suppressed the expression of ApoE/LDL-R at both doses.

There was no remarkable difference in neuronal MAP2 staining between the 3 groups.Conclusions: These findings suggest that both low and high doses of telmisartan prevented the activation of ApoE/LDL-R in SHR-SR after tMCAO, and that the antimetabolic effect was regarded as the most important mechanism with few additional benefitsby lowering BP in this transient stroke model.

Pe-003-5

Reducing Hemorrhagic Transformation by Rivaroxaban and Apixaban with tPA in Ischemic Stroke of Rat

1Department of Neurology, Okayama National Hospital Medical Center，

2Department of Neurology, Okayama University

○Syoichiro Kono¹，Toru Yamashita²，Kota Sato²，Nozomi Hishikawa²， Yasuyuki Ohta²，Yasuhiro Manabe¹，Koji Abe²

Background and Purpose:This study assesses the risks and benefits of tissue plasminogen activator (tPA) treatment under oral anticoagulation with new oral anticoagulants (NOACs) in transient middle cerebral artery occlusion (tMCAO) .Methods: After pretreatment with warfarin (0.2 mg/kg/day), rivaroxaban (2 mg/kg/day), apixaban (10mg/kg/day) or vehicle (0.5％ carboxymethyl cellulose sodium salt) for 7 days, tMCAO was induced for 120 min, followed by reperfusion and tPA (10 mg/kg/10 ml). At 24 hr after reperfusion, markers for the neurovascular unit at the peri-ischemic lesion were immunohistochemically examined in brain sections, and MMP-9 activity was measured by zymography.Results:Intracerebral hemorrhage volume was significantly improved in the NOACs-pretreated group compared with the warfarin-pretreated group. A marked dissociation between astrocyte foot processes and the basal lamina or pericyte was observed in the warfarin-pretreated group, which was greatly improved in the NOACs- pretreated group. Furthermore, a remarkable activation of MMP-9 in the ipsilateral warfarin-pretreated rat brain was greatly reduced in NOACs-pretreated rats.Conclusion:

The present study reveals that the mechanism of intracerebral hemorrhage with warfarin- pretreatment plus tPA in ischemic stroke rats is the dissociation of the neurovascular unit, including the pericyte. Neurovascular protection by NOACs could partially explain the reduction in hemorrhagic complication by NOACs reported from clinical study.

Pe-003-6

Effect of thrombin and cAMP-PKA signaling on the angiogenesis in brain microvasculature

Department of Neurology, Osaka University Graduate School of Medicine

○Hideaki Kanki，Tsutomu Sasaki，Akihiro Watanabe，Toshiro Yukami，

Manabu Sakaguchi，Hideki Mochizuki

[Objective] Angiogenesis play crucial roles in cerebrovascular disease including stroke for improving post stroke recovery coupled with neurogenesis. cAMP-PKA activity regulate angiogenic responses. Thrombin which is well known the factor involved in blood coagulation and is thought to act on endothelial cells has been also reported to play a pivotal role in the initiation of angiogenesis in cancer. Various discussion have been done about angiogenesis, but detailed mechanism remain incompletely understood, particularly under ischemic conditions. Therefore, we examined the effect of thrombin and cAMP-PKA on both angiogenesis and endothelial function. [Methods]

We used bovine brain microvascular endothelial cells (BBMC) . For evaluating angiogenesis, proliferation, tube formation and migration assay was performed. Tube formation assay with a Matrigel and wound migration assay were performed under serum deprivation or hypoxic conditions. Forskolin, p8CPT cAMP (cyclic AMP analogue), and thrombin was used. We have examined cAMP-mediated angiogenesis via CREB (cAMP response element binding protein), using CREB mutant (S133A, R314A, 133/314 double mutant) . [Results] BBMC plated on the Matrigel formed capillary like structures. Both Forskolin and p8CPT cAMP enhanced tube formation and migration. These effects were blocked by either pretreatment with H89 (PKA inhibitor) or CREB mutant. We have also examined angiogenic responses of BBMC in thrombin treatment. [Conclusions] In brain microvascular endothelial cells, cAMP- PKA-CREB pathway has essential role for angiogenesis.

- 481 -

18日

㈬ ポス ター

（英 語) Pe-003-1

TNF-α is associated with the mechanism for leptomeningeal arteriogenesis impairment in db/db mice

1Department of Neurology, Osaka University Graduate School of Medicine，²Department of Stroke Medicine, Kawasaki Medical School，³Department of Neurology, Tokyo Women’s Medical University

○Toshiro Yukami¹，Yoshiki Yagita²，Hideaki Kanki¹，Akihiro Watanabe¹，Naoki Oyama¹， Yasukazu Terasaki¹，Tsutomu Sasaki¹，Manabu Sakaguchi¹，Kazuo Kitagawa³， Hideki Mochizuki¹

[Objective] Leptomeningeal arteriogenesis is a key factor that defines the severity of ischemic stroke. Patients with stroke generally have vascular risk factors, such as diabetes. We found that leptomeningeal arteriogenesis 14 days after unilateral common carotid artery occlusion was impaired in db/db mice. This study was performed to examine the mechanism for leptomeningeal arteriogenesis impairment in db/db mice. [Methods] Macrophages play an important role in the process of arteriogenesis. We performed immunostaining using Mac-2 antibody to assess the degree of macrophage accumulation on the dorsal surface of the brain 7 days after CCA occlusion in db/+ and db/db mice. Next, we investigated the mRNA expression of several macrophage-related factors in the left cerebral cortex, including leptomeningeal anastomoses, in db/+ and db/db mice using real-time polymerase chain reaction. Finally, we tested whether the leptomeningeal arteriogenesis could be restored by pharmaceutical intervention in the db/db mice. [Results] The number of Mac-2-positive cells was increased and TNF-α mRNA expression was induced after common carotid artery occlusion in the db/+ mice.

However, these responses were not observed in the db/db mice. Administration of etanercept (TNF-α inhibitor) before common carotid artery occlusion restored the hypoperfusion-induced leptomeningeal arteriogenesis in db/db mice. [Conclusions] These results indicate that suppression of the TNF-α response to hypoperfusion is the major contributing factor for the leptomeningeal arteriogenesis impairment in db/db mice.

Pe-003-2

Heteroduplex oligonucleotide reduced gene expression in focal ischemic brain in mice

1Department of Neurology and Neurological Science, Tokyo Medical and Dental University，²Institute Multidisciplinary Research for Advanced Materials, Tohoku University

○Fuying Li¹，Satoru Ishibashi¹，Kotaro Yoshioka¹，Takehiko Wada²， Masahiko Ichijo¹，Eri Iwasawa¹，Jindong Song¹，Yongquan Zhang¹， Takanori Yokota¹

[Backgrounds]Tocopherol-conjugated heteroduplex oligonucleotide (Toc-HDO) exerts a strong gene silencing effect on many organs, but the delivery of Toc-HDO to the brain is very limited by the blood brain barrier (BBB). Since BBB is disrupted at the acute phase of stroke, intravenously administered (i.v.) Toc-HDO may further penetrate the BBB and reduce target gene expression primarily in ischemic region.[Methods]The permanent middle cerebral artery occlusion (pMCAO) was induced in male C57BL/6 mice (n=60). Toc-HDO targeting mMALAT1 was administrated via tail vein at 24 hours after pMCAO, and the MALAT1 gene expression of brain was quantified by RT-PCR. The distribution of fluorescence labeled Toc-HDO in the ischemic brain was investigated under confocal microscopy with cell specific markers.[Results]Three days after Toc-HDO administration, MALAT1 mRNA levels decreased in the ischemic region at 24 hours after pMCAO; the mMALAT1 levels in pMCAO+Toc-HDO group was significantly reduced to 30.1±9.5％ compared with pMCAO+PBS controls (p ＜ 0.01). Fluorescence labeled Toc-HDO distributed in the ischemic brain, primarily detected in neuronal and endothelial cells, but not in astrocytes. Total infarction volume was significantly larger in pMCAO+Toc-HDO group compared with controls.[Conclusion]We found a successful gene silencing effect of the ischemic brain by means of i.v. administration of Toc-HDO targeting mMALAT1, which exacerbates ischemic injury. Our findings establish proof of concept for the application of HDO to gene silencing therapy for acute stroke.

Pe-003-3

Serine racemase inhibition induces NO-mediated neurovascular protection during cerebral ischemia

Department of Neurology, Osaka University Graduate School of Medicine

○Akihiro Watanabe，Tsutomu Sasaki，Toshiro Yukami，Hideaki Kanki，

Manabu Sakaguchi，Hideki Mochizuki

[Purpose] There are no effective neuroprotectant drugs for acute cerebral ischemia. Serine racemase (SR) synthesizes D-serine, which is involved in N- methyl-D-aspartate (NMDA) receptor-induced neurotoxicity. However, regulatory mechanisms controlling SR-activity in the neurovascular unit during cerebral ischemia remain to be clarified. So, we investigated the effects of SR inhibition on neurovascular protection after ischemia.[Methods] We investigated whether the SR inhibitor phenazine methosulfate (PMS) could alleviate neuronal damage in an in vitro ischemic model (oxygen glucose deprivation [OGD]) using neuron cultures and in an in vivo mouse model of ischemia (middle cerebral artery occlusion [MCAO]) . We examined the effect of PMS on SR expression and phosphorylation by western blotting. Changes in cerebral blood flow (CBF) associated with administration of PMS were detected by laser speckle imaging.

Finally, we tested whether NO production was affected by PMS-administration after ischemia both in vitro and in vivo.[Results] PMS decreased cell death after OGD in neuron cultures. Infarct volumes were also reduced in PMS-treated mice.

PMS inhibited SR phosphorylation after ischemia. Reductions in regional CBF after MCA occlusion were improved by administration of PMS. Treatment with PMS increased both phosphorylation of eNOS and NO production.[Conclusions]

These findings indicate that SR inhibition acts as a neuroprotectant in the neurovascular unit and ameliorant of CBF abnormalities post-stroke. Thus, pharmacologic SR inhibition has potential clinical applications.

Pe-003-4

Telmisartan Treatment Strongly Improved ApoE/LDL-R Signals in Post-Stroke SHR-SR

Department of Neurology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences

○Toru Yamashita，Yun Zhai，Kota Sato，Mami Takemoto，

Nozomi Hishikawa，Yasuyuki Ohta，Koji Abe

Background: Telmisartan, an angiotensin receptor blocker also called metabosartan,is a promising solution for preventing cognitive decline or the incidence of dementia.Methods:We examined the effects of telmisartan on cholesterol transport- related proteins (apolipoprotein E (ApoE)/low-density lipoprotein receptor (LDL- R) ) and microtubule-associated protein 2 (MAP2) in the brain of spontaneously hypertensive stroke resistant (SHR-SR). SHR-SR received transient middle cerebral artery occlusion (tMCAO) for 90 minutes at 12 weeks of age and then was divided into 3 experiment groups including a vehicle, low-dose telmisartan (0.3 mg/kg/day), and high-dose telmisartan (3 mg/kg/day).Results: The low dose served to improve the metabolic syndrome of SHR-SR without lowering the blood pressure (BP) whereas the high dose was used to improve metabolic syndrome while lowering BP.

Immunohistologic analysis showed that ApoE expression of cortical neurons was strong in the vehicle group at 6, 12, and 18 months of age. On the other hand, LDL-R expression was transiently increased at 6 months of age only on the ipsilateral side.

Telmisartan dramatically suppressed the expression of ApoE/LDL-R at both doses.

There was no remarkable difference in neuronal MAP2 staining between the 3 groups.Conclusions: These findings suggest that both low and high doses of telmisartan prevented the activation of ApoE/LDL-R in SHR-SR after tMCAO, and that the antimetabolic effect was regarded as the most important mechanism with few additional benefitsby lowering BP in this transient stroke model.

Pe-003-5

Reducing Hemorrhagic Transformation by Rivaroxaban and Apixaban with tPA in Ischemic Stroke of Rat

1Department of Neurology, Okayama National Hospital Medical Center，

2Department of Neurology, Okayama University

○Syoichiro Kono¹，Toru Yamashita²，Kota Sato²，Nozomi Hishikawa²， Yasuyuki Ohta²，Yasuhiro Manabe¹，Koji Abe²

Background and Purpose:This study assesses the risks and benefits of tissue plasminogen activator (tPA) treatment under oral anticoagulation with new oral anticoagulants (NOACs) in transient middle cerebral artery occlusion (tMCAO) .Methods: After pretreatment with warfarin (0.2 mg/kg/day), rivaroxaban (2 mg/kg/day), apixaban (10mg/kg/day) or vehicle (0.5％ carboxymethyl cellulose sodium salt) for 7 days, tMCAO was induced for 120 min, followed by reperfusion and tPA (10 mg/kg/10 ml). At 24 hr after reperfusion, markers for the neurovascular unit at the peri-ischemic lesion were immunohistochemically examined in brain sections, and MMP-9 activity was measured by zymography.Results:Intracerebral hemorrhage volume was significantly improved in the NOACs-pretreated group compared with the warfarin-pretreated group. A marked dissociation between astrocyte foot processes and the basal lamina or pericyte was observed in the warfarin-pretreated group, which was greatly improved in the NOACs- pretreated group. Furthermore, a remarkable activation of MMP-9 in the ipsilateral warfarin-pretreated rat brain was greatly reduced in NOACs-pretreated rats.Conclusion:

The present study reveals that the mechanism of intracerebral hemorrhage with warfarin- pretreatment plus tPA in ischemic stroke rats is the dissociation of the neurovascular unit, including the pericyte. Neurovascular protection by NOACs could partially explain the reduction in hemorrhagic complication by NOACs reported from clinical study.

Pe-003-6

Effect of thrombin and cAMP-PKA signaling on the angiogenesis in brain microvasculature

Department of Neurology, Osaka University Graduate School of Medicine

○Hideaki Kanki，Tsutomu Sasaki，Akihiro Watanabe，Toshiro Yukami，

Manabu Sakaguchi，Hideki Mochizuki

[Objective] Angiogenesis play crucial roles in cerebrovascular disease including stroke for improving post stroke recovery coupled with neurogenesis. cAMP-PKA activity regulate angiogenic responses. Thrombin which is well known the factor involved in blood coagulation and is thought to act on endothelial cells has been also reported to play a pivotal role in the initiation of angiogenesis in cancer. Various discussion have been done about angiogenesis, but detailed mechanism remain incompletely understood, particularly under ischemic conditions. Therefore, we examined the effect of thrombin and cAMP-PKA on both angiogenesis and endothelial function. [Methods]

We used bovine brain microvascular endothelial cells (BBMC) . For evaluating angiogenesis, proliferation, tube formation and migration assay was performed. Tube formation assay with a Matrigel and wound migration assay were performed under serum deprivation or hypoxic conditions. Forskolin, p8CPT cAMP (cyclic AMP analogue), and thrombin was used. We have examined cAMP-mediated angiogenesis via CREB (cAMP response element binding protein), using CREB mutant (S133A, R314A, 133/314 double mutant) . [Results] BBMC plated on the Matrigel formed capillary like structures. Both Forskolin and p8CPT cAMP enhanced tube formation and migration. These effects were blocked by either pretreatment with H89 (PKA inhibitor) or CREB mutant. We have also examined angiogenic responses of BBMC in thrombin treatment. [Conclusions] In brain microvascular endothelial cells, cAMP- PKA-CREB pathway has essential role for angiogenesis.

- 482 -

18日

㈬ ポス ター

（英 語)

Pe-003-7

A Quantitative proteomic analysis of mesenchymal stem cell transplantation on ischemic stroke rats

1Department of Neurology, Shimane University School of Medicine，

2Department of Laboratory Medicine, Shimane University School of Medicine，

3Department of Biosignaling and Radioisotope Experiment, Interdisciplinary Center for Science Research, Organization for Research, Shimane University

○Shingo Mitaki¹，Atsushi Nagai²，Kazumi Satoh³，Sheikh Abdullah²， Keiichi Onoda¹，Ken-ichi Matsumoto³，Shuhei Yamaguchi¹

[Introduction] Previously, we demonstrated that transplantation of a human mesenchymal stem cell (B10) decreased lesion size and neurological deficits accompanied by decreased expression of proinflammatory factors in a rat model of transient middle cerebral artery occlusion (MCAO). Given the multipotency of B10, other mechanisms of B10 in transplantation-induced neurological improvement may exist. The purpose of this study was to identify key proteins involved in the therapeutic effects of B10 transplantation using a multiplex proteomics approach.[Methods] One day after MCAO, PBS or B10 was transplanted intravenously (5 rats, respectively). Seven days after transplantation, a proteomic approach was performed using isobaric tagging and peptide quantification via matrix-assisted laser desorption/ionisation tandem mass spectrometry. [Results]

Twelve proteins in core lesions and 16 proteins in penumbra lesions were found to be differentially regulated following B10 transplantation. In core lesions, 2 proteins (GFAP and Alb) which were overexpressed after PBS injection were significantly down-regulated. In contrast, in penumbra lesions, 3 proteins (EAAT2, neurofilament, and CNPase) were significantly up-regulated. Based on these results, in core lesions, it is possible that B10 transplantation preserves the integrity of the blood-brain barrier (Alb) and in penumbra lesions, suppresses neuronal excitotoxicity (EAAT2), inflammation (CNPase), and increased axonal growth (neurofilament). [Conclusions] This study highlights the pleiotropic actions of B10 for stroke treatment.

Pe-004-1 withdrawn

Pe-004-2

Regional cerebral blood flow patterns associated with depression in early AD and DLB

1Department of Geriatric medicine, Tokyo Medical University Hospital，

2Department of Ultrahigh Field MRI, Institute for Biomedical Sciences, Iwate Medical University

○Kentaro Hirao¹，Soichiro Shimizu¹，Fumio Yamashita²， Tomohiko Sato¹，Hidekazu Kanetaka¹，Takahiko Umahara¹， Hirofumi Sakurai¹，Haruo Hanyu¹

Objectives: To investigate the relationship between depression and regional cerebral blood flow (rCBF) patterns in Alzheimer disease (AD) and dementia with lewy bodies (DLB).

Methods: We performed neuropsychological test, brain MRI and SPECT (I-IMP) of early stage of probable AD (n=25) and probable DLB (n=16). We excluded the subjects with cerebral infarctions and/or severe white matter abnormalities (Fazekas score 3). The voxel-wise correlations of rCBF with geriatric depression scale (GDS) score (full points are 15) was performed in each group, respectively using SPM8. The correlations were reported at p=0.01, spatial extent threshold 50 voxels.Results: The mean PVH/WMH score of Fazekas scale was 1.1/1.6 in AD and 1.1/1.2 in DLB. The mean GDS score was 5.7 in AD and 6.9 in DLB. There were no significant differences of PVH/WMH and GDS score between 2 group. The regression analysis showed that there was a significant negative correlation of rCBF of lt.inferior frontal, lt.medial frontal, lt.superior frontal gyrus, lt.rectal gyrus, rt.anterior cingulate, lt.inferior parietal lobule with GDS score in AD, while in more extent frontal area(BA47,BA6,BA9,BA46,BA10,BA8), rt.insula, rt.inferior temporal gyrus and rt.superior occipital gyrus in DLB.Conclusions: There was some different rCBF patterns associated with depression between AD and DLB, which suggests that neurobiological mechanisms associated with depression might be different in each group.

Pe-004-3

How we can predict ADL-decline from neuropsychological test in early-onset Alzheimer’s disease

Department of Neurology, JCHO Tokyo Takanawa Hospital

○Manabu Tsumoto

[OBJECTIVE] To examine the predictive factors of ADL-decline in early-onset Alzheimer’s disease patients, whose MMSE scores show few changes at follow- up, we use multiple regression analysis of the neuropsychological test battery.

[METHODS] The study was approved by our hospital research ethics committee.

Four patients (mean age 61.5±4.1) who first visited our hospital and diagnosed as early-onset Alzheimer’s disease participated in the study. Their test score was as follows: MMSE 25.8 ± 3.2, Montreal Cognitive Assessment (MoCA) 20.0 ± 3.3, Addenbrooke’ s Cognitive Examination Revised (ACE-R) 79.5 ± 6.9, Frontal Assessment Battery (FAB)13.8±1.7, and Rivermead Behavioural Memory Test (RBMT) SPS 6.5 ± 2.9, SS 2.3 ± 1.0. The Functional Activities Questionnaire (FAQ) measured for instrumental activities of daily living was 4.8±3.0. After one year follow-up, we calculated their test score change. [RESULTS] ΔRBMT SS (R2 = 0.474) could be useful as an explanatory variable for FAQ decline in comparison with Δ MMSE (R2 = 0.356) and others. [CONCLUSIONS] It was suggested that RBMT is helpful for following-up in early-onset Alzheimer’ s disease patients.

Pe-004-4

HLA-A2 alleles mediate Alzheimer’s disease by altering hippocampal volume in ADNI subjects

Qingdao Municipal Hospital

○Zixuan Wang，Huifu Wang，Lin Tan，Jintai Yu，Lan Tan Objective:HLA-A alleles have been shown to be associated with Alzheimer’s disease (AD) . In this study, we firstly investigated the association of gene variants in HLA-A and brain structure on magnetic resonance imaging (MRI) in the Alzheimer’s Disease Neuroimaging Initiative (ADNI) subjects to explore the effects of HLA-A on AD.Methods:We included 712 subjects from the ADNI dataset and evaluated the impact of the HLA-A loci and haplotype on the brain atrophy of hippocampus, parahippocampus, posterior cingulate, precuneus, middle temporal, entorhinal cortex and amygdala on MRI in a multiple linear regression model. Eight SNPs in HLA-A were identified in the dataset and passed quality control.Results:In hybrid population analysis, we found a marginally significant association between rs9260168 and the atrophy of the left parahippocampus (Pc=

0.054), rs3823342 and the atrophy of the left parahippocampus (Pc = 0.054), rs76475517, which only exists in Caucasians with HLA-A23 or HLA-A24 alleles, and the atrophy of the right amygdala (Pc=0.085) at baseline. In particular, the haplotype (TGACAAGG), as a surrogate marker of HLA-A2, was founded to be associated with the baseline atrophy of the right hippocampus (P = 0.047) . Furthermore, we detected the above four associations in mild cognitive impairment (MCI) sub-population analysis. Conclusion: Our study provided preliminary evidences supporting HLA-A2 in Caucasians contribute to the risk of AD by modulating the alteration of right hippocampal volume.

Pe-005-1

Intravenous sedation assisted dental treatment on patients with dementia

1Department of Neurology, Shuang-Ho Hospital, Taipei Medical University，

2Department of Dentistry, Shuang-Ho Hospital, Taipei Medical University

○Yao-hsien Huang¹，Ta-San Huang²，En-Sheng Ko²，Pung-Fei Tsai²， Shu-Hsien Huang²

ObjectivePeople with dementia are susceptible to dental diseases. Abundant clinical evidences show that oral health deteriorates as the severity of dementia progresses.

However, dental treatment is challenging due to the progressive cognitive impairment. Our study assesses the efficacy and tolerability of intravenous sedation (IVS) on patients with dementia undergoing dental treatment, compared to non- dementia patients.MethodsPatients with special needs who have undergone dental treatment controlled with IVS between June 2013 to May 2014 were included in our cohort.ResultsA total of 293 patients received dental treatment IVS-assisted with propofol. Of these, 122 were patients with multiple disabilities, 75 patients with mental disabilities, and 43 with autism spectrum disorders. Only 14 of 293 patients were diagnosed with dementia. Patients with dementia undergoing dental treatment assisted with IVS required higher doses of sedative drugs. In terms of tolerability, the most commonly encountered complication included inadequate sedation, cough and hypertension.ConclusionsPatients with moderate to severe dementia undergoing dental treatment can be properly managed with IVS. The main advantages include easy induction of sedation, smooth handling during dental management, and shorter recovery time. However, a higher dose of sedatives might be required in patients with dementia. Pre-operative evaluation of comorbidities as well as concomitant medications is of utmost importance to avoid undesirable drug-drug interaction, and unwanted complications during the procedure.