21日
㈯ ポス ター
（英 語) Pe-053-1
Pe-053-2 WIthdrawn
Pe-053-3 Withdrawn
Pe-053-4
Effects of Tranexamic Acid on Hematoma Volume in Patients with Hypertensive Intracerebral Hemorrhage
Iloilo Mission Hospital
○Ma. Daisy P. Tabuena，Kay Hazell E. Duran，Shake G. Lumaque Background:Intracerebral hemorrhage accounts for 10-15％ of all strokes.It has the highest mortality with a 1-year survival rate of ＜ 50％．Hematoma volume is a critical determinant of outcome in these patients.Objectives:To determine the effect of Tranexamic Acid on intracerebral hematoma volume in patients with hypertensive intracerebral hemorrhage admitted at our institution.To compare the change from baseline in hematoma volume between patients given Tranexamic acid + standard management and patients given standard management alone.Methods: This is a retrospective cross sectional study conducted among patients admitted at our institution from January 2012 to December 2014 with a diagnosis of Intracerebral Hemorrhage confirmed by cranial CT scan.Medical records were reviewed and patients were grouped as to whether they received tranexamic acid + standard management and standard management alone.Hematoma volume at baseline and follow-up were determined.The change in hematoma volume between the 2 groups were compared using the paired t test.Results:There were more male patients,age range is from 29-87 years old. Patients given tranexamic acid had significant reduction in hematoma volume from baseline (39.8ml to 28.7ml) compared to patients not given tranexamic acid 43.08ml to 43.91m).The difference in volume reduction between the 2 groups was statistically significant with a p value of .005.Conclusion: Tranexamic acid when added to standard therapy reduces hematoma volume in patients with hypertensive Intracerebral Hemorrhage.
Pe-053-5
PREDICTORS OF SIGNIFICANT CAROTID STENOSIS AMONG PATIENTS UNDERGOING CAROTID DUPLEX ULTRASONOGRAPHY
The Medical City, Philippines
○Maria Angelica M. Geronimo，Artemio A. Roxas
Background:Carotid duplex scan (CDS) is a non-invasive screening tool used to document carotid artery stenosis (CAS). Factors such as costs, lower prevalence of significant CAS among Asians, and disproportionate number of patients who undergo revascularization techniques are considered in the selection of ideal patients that would benefit from performing CDS. The study aimed to determine the clinical predictors that would result to findings of a significant CAS, defined as 50％ and above.Methods:This is a cross-sectional observational study conducted in a tertiary hospital. CDS results of all consecutive patients from a total of 5 years were retrieved. Non-Asian patients were excluded in this study. Statistical analysis of clinical indicators used the Mann-Whitney U Test while risk ratio calculations used Chi Square.Results:A total of 3,717 CDS were retrieved from all consecutive patient records. There was a low rate of 8％ of significant findings of CAS (moderate CAS 6％，severe CAS 1.3％，and total occlusion 1.2％) among these patients. Patients aged 50-69 years old; female; with prior history of stroke, AF/dysrhythmia, and dyslipidemia; and presenting weakness, numbness, slurred speech, and headache in the clinic are likely to have findings of significant CAS on CDS and should therefore undergo CDS.Conclusion:In clinical practice, CDS is an over-utilized tool. The study population have a low rate of significant CAS findings on CDS. Therefore, CDS as a relatively expensive tool should be reserved only for patients likely to have significant CAS.
Pe-053-6
Men with primary OSA have a higher degree of a Neurological Condition than women of same age group
EENA Comprehensive Neurology and Sleep Center
○Edith Mensah-osman，Nadir Ishag-Osman
INTRODUCTION:Obstructive sleep apnea (OSA) affects a majority of the population and is associated with hypertension, obesity, diabetes, and an increased risk of heart attack and stroke. The overall objective of this study is to conduct a 2 year review and analysis of over 1000 patients in other to evaluate the comorbid conditions associated with the primary diagnosis of obstructive sleep apnea.METHODS: We identified and diagnosed patients with OSA using polysomnography and determined the prevalence of a secondary neurological condition at initial diagnosis and/or after CPAP treatment.KEY RESULTS:
Approximately 2-3 new patients present daily and are subsequently diagnosed with obstructive sleep apnea. Majority of the newly diagnosed OSA patients present with secondary co-morbid pathology. In men over 55 years of age the primary diagnosis of OSA is associated with a secondary neurological condition, whereas there was a greater variation in women of the same age group.CONCLUSIONS:OSA is associated with neurological conditions in middle-aged men compared to women. Ongoing analysis is focused on other factors that may cause this predisposition in men compared to women, and evaluate if there is a direct link between neurological conditions and the development of OSA.
Withdrawn
539
-21日
㈯ ポス ター
（英 語) Pe-053-1
Pe-053-2 WIthdrawn
Pe-053-3 Withdrawn
Pe-053-4
Effects of Tranexamic Acid on Hematoma Volume in Patients with Hypertensive Intracerebral Hemorrhage
Iloilo Mission Hospital
○Ma. Daisy P. Tabuena，Kay Hazell E. Duran，Shake G. Lumaque Background:Intracerebral hemorrhage accounts for 10-15％ of all strokes.It has the highest mortality with a 1-year survival rate of ＜ 50％．Hematoma volume is a critical determinant of outcome in these patients.Objectives:To determine the effect of Tranexamic Acid on intracerebral hematoma volume in patients with hypertensive intracerebral hemorrhage admitted at our institution.To compare the change from baseline in hematoma volume between patients given Tranexamic acid + standard management and patients given standard management alone.Methods: This is a retrospective cross sectional study conducted among patients admitted at our institution from January 2012 to December 2014 with a diagnosis of Intracerebral Hemorrhage confirmed by cranial CT scan.Medical records were reviewed and patients were grouped as to whether they received tranexamic acid + standard management and standard management alone.Hematoma volume at baseline and follow-up were determined.The change in hematoma volume between the 2 groups were compared using the paired t test.Results:There were more male patients,age range is from 29-87 years old. Patients given tranexamic acid had significant reduction in hematoma volume from baseline (39.8ml to 28.7ml) compared to patients not given tranexamic acid 43.08ml to 43.91m).The difference in volume reduction between the 2 groups was statistically significant with a p value of .005.Conclusion: Tranexamic acid when added to standard therapy reduces hematoma volume in patients with hypertensive Intracerebral Hemorrhage.
Pe-053-5
PREDICTORS OF SIGNIFICANT CAROTID STENOSIS AMONG PATIENTS UNDERGOING CAROTID DUPLEX ULTRASONOGRAPHY
The Medical City, Philippines
○Maria Angelica M. Geronimo，Artemio A. Roxas
Background:Carotid duplex scan (CDS) is a non-invasive screening tool used to document carotid artery stenosis (CAS). Factors such as costs, lower prevalence of significant CAS among Asians, and disproportionate number of patients who undergo revascularization techniques are considered in the selection of ideal patients that would benefit from performing CDS. The study aimed to determine the clinical predictors that would result to findings of a significant CAS, defined as 50％ and above.Methods:This is a cross-sectional observational study conducted in a tertiary hospital. CDS results of all consecutive patients from a total of 5 years were retrieved. Non-Asian patients were excluded in this study. Statistical analysis of clinical indicators used the Mann-Whitney U Test while risk ratio calculations used Chi Square.Results:A total of 3,717 CDS were retrieved from all consecutive patient records. There was a low rate of 8％ of significant findings of CAS (moderate CAS 6％，severe CAS 1.3％，and total occlusion 1.2％) among these patients. Patients aged 50-69 years old; female; with prior history of stroke, AF/dysrhythmia, and dyslipidemia; and presenting weakness, numbness, slurred speech, and headache in the clinic are likely to have findings of significant CAS on CDS and should therefore undergo CDS.Conclusion:In clinical practice, CDS is an over-utilized tool. The study population have a low rate of significant CAS findings on CDS. Therefore, CDS as a relatively expensive tool should be reserved only for patients likely to have significant CAS.
Pe-053-6
Men with primary OSA have a higher degree of a Neurological Condition than women of same age group
EENA Comprehensive Neurology and Sleep Center
○Edith Mensah-osman，Nadir Ishag-Osman
INTRODUCTION:Obstructive sleep apnea (OSA) affects a majority of the population and is associated with hypertension, obesity, diabetes, and an increased risk of heart attack and stroke. The overall objective of this study is to conduct a 2 year review and analysis of over 1000 patients in other to evaluate the comorbid conditions associated with the primary diagnosis of obstructive sleep apnea.METHODS: We identified and diagnosed patients with OSA using polysomnography and determined the prevalence of a secondary neurological condition at initial diagnosis and/or after CPAP treatment.KEY RESULTS:
Approximately 2-3 new patients present daily and are subsequently diagnosed with obstructive sleep apnea. Majority of the newly diagnosed OSA patients present with secondary co-morbid pathology. In men over 55 years of age the primary diagnosis of OSA is associated with a secondary neurological condition, whereas there was a greater variation in women of the same age group.CONCLUSIONS:OSA is associated with neurological conditions in middle-aged men compared to women. Ongoing analysis is focused on other factors that may cause this predisposition in men compared to women, and evaluate if there is a direct link between neurological conditions and the development of OSA.
Withdrawn
540
-21日
㈯ ポス ター
（英 語)
Pe-053-7
Challenges of a Comprehensive Neurology & Sleep Center-from Setup to Reimbursements
EENA Comprehensive Neurology and Sleep Center
○Mensah-osman Edith
Introduction: Setting up a solo medical private practice is a nightmare that physicians dare not venture. Most will rather join a practice for the first time after training, or from academia, since we are not sufficiently taught to manage the nuances of this new arena. Indeed, even an MBA geared towards executive healthcare management did not prepare for the challenges encountered.Methods:Layout of initial concepts;
Update of concepts from practical use, Modification/Change of concepts from experience -"Building the Bridge As you Walk On It"-Robert E. Quinn.Key Results:
Define practice model;Determinelocation;DesignEMR/PM;Developstandard operating protocols;Negotiatecontracts; Recognizeissues with reimbursements;
CreateCE programs;Participatein community outreach;Attainrecognition and accreditation;Recruityour staff;Identifyoffice bottlenecks;Researchprograms;
Marketyour niche.Conclusions:Success is not solely a function of reportable income tax but a combination of tangible and intangible financial attributes. Respect by colleagues, patients and staff are the best marketing tools in the first two years.
Participation in research programs can be the "life-line" needed to stay "afloat" in this period.
Pe-054-1
Validation of the Filipino Version of National Institute of Health Stroke Scale
1Philippine General Hospital，2Critical Care Nurses Association of the Philippines, Inc.○Ela A. Barcelon1，Mari Anne Kristin Dale N. Moll1，Diana J. Serondo2， Ma. Epifania V. Collantes1
Significance & Objective: Several countries have translated and culturally adapted the English NIHSS to their native language. This study aimed to validate the recently translated and culturally adapted Filipino version of NIHSS.Methods:
Using consecutive sampling, two independent NIHSS certified raters administered the Fil-NIHSS to 53 stroke patients, with informed consent, consulting the emergency and outpatient department in a tertiary hospital. The Fil-NIHSS were administered within 24 hours from the administration of the English NIHSS. Correlation between the English and Filipino NIHSS was measured using Pearson’ s Product-Moment Correlation and presented with a scatterplot. Validity of the Fil-NIHSS were tested using paired t-test. Internal consistency of the questionnaire was measured using Cronbach’s alpha while the inter-rater reliability were measured and tested using Cohen’s kappa test. The level of significance was set at 0.05.Results and Conclusion: The high correlation (0.9451) to English NIHSS, validity and excellent internal consistency (α= 0.91) of the Fil-NIHSS makes it suitable to be administered to Filipino-speaking patients who are unable to understand the English language. The low interrater reliability may imply, the need to construct a formal course training for Fil-NIHSS certification. Nevertheless, while we still do not have such, the Fil-NIHSS is a valid and reliable stroke scale that can administered to Filipino-speaking stroke patients.
Pe-054-2 withdrawn
Pe-054-3
Pattern and Risk Factors of Stroke in a District Hospital of Johor, Malaysia
1Department of Medicine, Hospital Enche’ Besar Hajjah Khalsom, Kluang, Johor, Malaysia，2Department of Medicine,Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia
○Yang Liang Boo1，Hong Tak Lim1，Chee Ming Goh1，
Nai Chien Huan1，Pek Woon Chin1，Fan Kee Hoo2，Siew Mooi Ching2， Hamidon Basri2
AimStroke is one of the leading causes of death and disability in developing countries.
Epidemiology data on acute stroke is very limited in Malaysia. We aimed to explore the demographic data, risk factors and patterns of stroke in a Malaysia district hospital.MethodsAn observation study was conducted in a district hospital from July 2014 to December 2014. Risk factors, clinical characteristics and details of the acute stroke were analyzed.Results143 patients were recruited with mean age of 62±14 years old. 61.5％ were male with 70.6％ had primary education level and below. Risk factors identified were hypertension(76.2％), diabetes (51.0％), dyslipidaemia(18.2％), prior history of stroke or TIA(18.6％), atrial fibrillation(8.4％), smoking(26.3％) and family history of stroke(1.4％). CT brain showed infarction in 69(48.3％) patients, hemorrhagic in 23 patients(16.1％) and normal in 50(35％) patients. The median hospital arrival time was 13.6 hours (IQR: 13.5 hours) with 43(30.1％) patients arrived hospital less than 4.5 hours. The median time of the door to CT scanning was 95 minutes (IQR: 132 minutes). The mean NIHSS score was 8.7. 82(52.7％) patients had small-vessel occlusion disease while 28(19.6％) patients were large artery disease based on TOAST classification.Conclusion With one-third of the patients arriving the hospital within 4.5 hours, the option of thrombolysis as the mainstay of treatment in acute ischemic stroke needs to be explored in a district hospital.
Public awareness on signs and symptoms of stroke is important to promote the eligibility of thrombolysis therapy.
Pe-054-4
Patient-level and hospital-level determinants of rehabilitation during acute stroke care in Taiwan
1Neurological Center, Cardinal Tien Hospital, New Taipei City, Taiwan，2School of Medicine, College of Medicine, Fu-Jen Catholic University, Xinzhuang District, New Taipei City, Taiwan
○Vinchi Wang1,2，Tzu-Hao Chao1，Tsung-Tai Chen2
Objectives: During acute stroke care, rehabilitation usage may be influenced by patient- and hospital-related factors. We would like to identify patient- and hospital-level determinants of population-level inpatient rehabilitation usage associated with acute stroke care.Methods: From data obtained from the claim information from the National Health Insurance Administration (NHIA) in Taiwan (2009-2011), we enrolled 82, 886 stroke patients with intracerebral hemorrhage and cerebral infarction from 207 hospitals. A generalized linear mixed model (GLMM) analyses with patient-level factors specified as random effects were conducted (for cross-level interactions) .Results: The rate of rehabilitation usage was 51％ during acute stroke care. The hospital-related factors accounted for a significant amount of variability (intraclass correlation, 50％) . Hospital type was the only significant hospital-level variable and can explain the large amount of variability (58％) . Patients treated in smaller hospitals experienced few benefits of rehabilitation services, and those with surgery in a smaller hospital used fewer rehabilitation services. All patient-level variables were significant.Conclusions:With GLMM analyses, we identified the hospital type and its cross-level interaction, and explained a large portion of variability in rehabilitation for stroke patients in Taiwan.
Pe-054-5 Withdrawn
21日
㈯ ポス ター
（英 語) Pe-054-6
WIthdrawn
Pe-055-1
Establishment of 6-OHDA induced-parkinsonian model rat with chimeric GFP+
bone marrow
Department of Neurology, School of Medicine, Sapporo Medical University
○Syuuichirou Suzuki，Hiromi Suzuki，Mai Fujikura，Tatsuo Manabe，
Akihiro Iwahara，Akihiro Matsumura，Takashi Matsushita，
Shin Hisahara，Jun Kawamata，Shun Shimohama
[Background] We previously reported the therapeutic effects of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) in a 6-OHDA induced-parkinsonian rat model via modulation of microglial activation. However, survival of the transplanted hBM-MSCs was not detected at the nigrostriatal sections of lesioned rats. We therefore thought that the intravenous injected-hBM-MSCs inhibit immunocytes in the blood vessel to infiltrate into the brain. To test the hypothesis, we produced transplanted green fluorescent protein-positive bone marrow cells (GFP-BMCs) into 6-OHDA induced-parkinsonian rats to distinguish microglia from macrophages which are sharing similar gene expression profiling. [Methods] We obtained GFP-BMCs from tibias and femurs of GFP-Tg rats. 5-week-old SD rats (n=9) were received whole-body irradiation with lead head caps. Subsequently, GFP-BMCs obtained by the above method (5.0 × 107cells/animal) were injected into the femoral vein of the irradiated rats. Four weeks after transplantation, 6-OHDA was transcranially injected into the left striatum. GFP-BMCs-transplanted rats were administered methamphetamine to count the number of rotations for 120 min. Behavioral analyses were performed on 7, 14, 21 and 28 days after 6-OHDA administration.[Results] GFP-BMCs-transplanted rats did not reduce the number of rotations compared to non-transplanted rats.
[Conclusions] We established 6-OHDA induced-parkinsonian model rat with chimeric GFP + BM, which would elucidate therapeutic mechanism of hBM-MSCs transplantation.
Pe-055-2
Expression of mRNA for adenosine A2A receptor in rat model of levodopa-induced dyskinesia
Department of Neurology, Kansai Medical University
○Mitsuaki Oki，Satoshi Kaneko，Satoshi Morise，Masataka Nakamura，
Reika Wate，Norihiro Takenouchi，Hirofumi Kusaka
Purpose It have been considered continuous dopaminergic stimulation is important to reduce dyskinesia induction in Parkinson’s disease (PD) treatment.
To investigate the difference according to the mode of levodopa administration, we analyzed mRNA expression of dopaminergic and non-dopaminergic receptors in relation to levodopa-induced dyskinesia (LID) . Methods 6-OHDA was stereotaxically injected into the unilateral medial forebrain bundle of SD rats to make unilateral PD model rats. These rats were subdivided into three groups and treated as follows; 1) no medication (N), 2) continuous levodopa infusion (C), and 3) intermittent levodopa injection (I). Two weeks after the treatment, severity of LID was scored. Expression of mRNA on the operated side was compared to that on the unoperated side by real time RT-PCR.ResultsLID was observed in group I. Both D1 and D2 receptor mRNAs were elevated in group I, but only D2 receptor mRNA expression was elevated in group C. Enkephalin mRNA in group C and dynorphin mRNA in group I were elevated, indicating that the direct pathway is conversely dominant over the indirect pathway in LID. Adenosine A2A receptor mRNA was elevated and enkephalin mRNA was not elevated in group I.
Therefore, elevated A2A receptor may be in association with LID expression through inhibiting the indirect pathway.ConclusionContinuous or intermittent mode of levodopa administration affects dopaminergic and non-dopaminergic receptor expression. Elevated A2A receptor expression by intermittent levodopa administration may be related to LID pathogenesis.
Pe-055-3
Temporarily restricted expression of alpha-synuclein using Tet-off system to establish PD mice model
1Department of Neurology, Neurological Institute, Graduate school of Medical Sciences, Kyushu University，2Department of Neuropsychiatry, School of Medicine, Keio University
○Hiroo Yamaguchi1，Atsushi Fujita1，Hayato Une1，Yinan Zhao1， Kenji Tanaka2，Jun-ichi Kira1
Object: Various animal models, in which α-synuclein is overexpressed in DA neurons, have been tried for Parkinson’s disease (PD) animal model. For true animal models of PD，α-synuclein should be overexpressed in DA neurons not in the developmental stage but rather in the adult stage. Among several inducible transgene systems, Tet-off system is more efficacious for regulating gene expression in a temporally restrictive manner. To investigate the molecular mechanism of DA neuron death related to α-synuclein pathology and to establish PD mice model, we aimed to establish the mice, in which α-synuclein is overexpressed in DA neurons in a temporarily restrictive manner using Tet-off system.Methods: We used TetO-α -SynA53TTg/+ mice and DAT-tTATg/+ mice. TetOα -SynA53TTg/+mice express mutant A53T human α -synuclein (α -SynA53T) regulated by a tetracycline operator. DAT-tTATg/+mice express a tetracycline-controlled transactivator protein (tTA) driven by the mouse dopamine transporter (DAT) promoter. We mated TetOα -SynA53TTg/+mice with DAT-tTATg/+mice and analyzed their offspring including TetOα -SynA53TTg/+; DAT-tTATg/+, TetO-α -SynA53TTg/+; DAT-tTA+/+, TetO-α -SynA53T+/+; DAT-tTATg/+, and TetO-α-SynA53T+/+;DAT-tTA+/+mice.Results and conclusion: Immunohistochemical analysis showed that TetO-α-SynA53TTg/+;DAT-tTATg/+mice successfully expressed α-SynA53T in DA neurons in the SN in the absence of doxycycline (Tet-off). We will analyze these mice whether they show DA neurons loss. TetO-α-SynA53TTg/+;DAT-tTATg/+mice are useful for temporarily restricted expression of α-synuclein.
Pe-055-4
Can exercise modify Parkinson’s disease?-using a new mouse model of Parkinson’s
disease-Department of Neurology, Graduate School of Medicine, Kyoto University
○Shinya Okuda，Norihito Uemura，Ryosuke Takahashi
[Background and Purpose] It has been shown that exercise improve the cognitive dysfunction and memory loss. In terms of the molecular mechanisms, exercise affects the expression of brain-derived neurotrophic factor (BDNF) through the PGC-1α/FNDC5 pathway in the hippocampus. Several clinical data showed that exercise is also effective for Parkinson’s disease, but the molecular mechanisms are unknown. Our purpose of this research is to validate the disease modifying effect of exercise on Parkinson’s disease, using a new mouse model. [Methods]
First, to establish new disease model, we injected α-Synuclein (α-Syn) fibrils into the striatum of the α-Syn BAC (bacterial artificial chromosome) transgenic mouse that overexpress α-Syn. We’ll analyze the α-Syn-positive Lewy Body pathology and the number of dopaminergic neurons and evaluate their motor function one, three, and six months after injection. Further, we plan to force these model mice to exercise with running wheels and to evaluate whether their pathology and motor function can be improved or not. [Result] In preliminary experiments, injection of α-Syn fibrils caused the propagation of α-Syn-positive Lewy Body pathology. [Conclusion] We’ve confirmed that inoculation of α-Syn fibrils induce Lewy Body pathology. Hereafter, we begin to establish the new disease model.
Pe-055-5
Pathological role of GBA2 in GBA1-deficient neuronopathic Gaucher’s disease model of medaka
1Department of Neurology, Graduate School of Medicine, Kyoto University，
2Laboratory for Molecular Membrane Neuroscience, RIKEN Brain Science Institute，3Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University，4Department of radiation Genetics, Graduate School of Medicine, Kyoto University
○Etsuro Nakanishi1，Norihito Uemura1，Hisako Akiyama2， Masato Kinoshita3，Hodaka Yamakado1，Shunichi Takdeda4， Yoshio Hirabayashi2，Ryosuke Takahashi1
[Objective] Recent genetic studies have identified heterozygous mutations in the GBA1 gene as a strong risk factor for sporadic Parkinson’s disease (PD). The GBA1 mutations are responsible for Gaucher’s disease (GD). We have reported that GBA1 knock-out (KO) medaka can survive long enough for pathological analysis of disease progression in contrast to the perinatal death of GBA1 KO mice. This neuronopathic GD medaka model displays α-synuclein accumulation in the brains and has a potential to investigate the mechanisms of α-synuclein accumulation in GD and GBA1-related PD. The non-lysosomal β-Glucosidase (GBA2), which is localized at the endoplasmic reticulum and Golgi apparatus, also cleaves glucosylceramide to glucose and ceramide. A recent study has reported that the deletion of GBA2 rescues the visceral manifestation of type1 GD mice model. To date, the relationship between GBA1 and GBA2 in the central nervous system in GD and GBA1-related PD remains unclear. Our objective is to clarify the pathological role of GBA2 in the central nervous system in GD and GBA1-related PD. [Methods] We planned to generate GBA2 KO medaka by clustered regularly interspaced short palindromic repeat (CRISPR) / CRISPR-associated nuclease (Cas9) system. [Results] We have successfully generated GBA2 KO medaka by CRISPR / Cas9.
GBA2 KO medaka appear to show no developmental abnormalities compared to wild-type medaka. [Conclusions] We will cross GBA2 KO medaka with GBA1 KO medaka to examine the genetic interaction between GBA1 and GBA2 in GD and GBA1-related PD.
541
-21日
㈯ ポス ター
（英 語) Pe-054-6
WIthdrawn
Pe-055-1
Establishment of 6-OHDA induced-parkinsonian model rat with chimeric GFP+
bone marrow
Department of Neurology, School of Medicine, Sapporo Medical University
○Syuuichirou Suzuki，Hiromi Suzuki，Mai Fujikura，Tatsuo Manabe，
Akihiro Iwahara，Akihiro Matsumura，Takashi Matsushita，
Shin Hisahara，Jun Kawamata，Shun Shimohama
[Background] We previously reported the therapeutic effects of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) in a 6-OHDA induced-parkinsonian rat model via modulation of microglial activation. However, survival of the transplanted hBM-MSCs was not detected at the nigrostriatal sections of lesioned rats. We therefore thought that the intravenous injected-hBM-MSCs inhibit immunocytes in the blood vessel to infiltrate into the brain. To test the hypothesis, we produced transplanted green fluorescent protein-positive bone marrow cells (GFP-BMCs) into 6-OHDA induced-parkinsonian rats to distinguish microglia from macrophages which are sharing similar gene expression profiling. [Methods] We obtained GFP-BMCs from tibias and femurs of GFP-Tg rats. 5-week-old SD rats (n=9) were received whole-body irradiation with lead head caps. Subsequently, GFP-BMCs obtained by the above method (5.0 × 107cells/animal) were injected into the femoral vein of the irradiated rats. Four weeks after transplantation, 6-OHDA was transcranially injected into the left striatum. GFP-BMCs-transplanted rats were administered methamphetamine to count the number of rotations for 120 min. Behavioral analyses were performed on 7, 14, 21 and 28 days after 6-OHDA administration.[Results] GFP-BMCs-transplanted rats did not reduce the number of rotations compared to non-transplanted rats.
[Conclusions] We established 6-OHDA induced-parkinsonian model rat with chimeric GFP + BM, which would elucidate therapeutic mechanism of hBM-MSCs transplantation.
Pe-055-2
Expression of mRNA for adenosine A2A receptor in rat model of levodopa-induced dyskinesia
Department of Neurology, Kansai Medical University
○Mitsuaki Oki，Satoshi Kaneko，Satoshi Morise，Masataka Nakamura，
Reika Wate，Norihiro Takenouchi，Hirofumi Kusaka
Purpose It have been considered continuous dopaminergic stimulation is important to reduce dyskinesia induction in Parkinson’s disease (PD) treatment.
To investigate the difference according to the mode of levodopa administration, we analyzed mRNA expression of dopaminergic and non-dopaminergic receptors in relation to levodopa-induced dyskinesia (LID) . Methods 6-OHDA was stereotaxically injected into the unilateral medial forebrain bundle of SD rats to make unilateral PD model rats. These rats were subdivided into three groups and treated as follows; 1) no medication (N), 2) continuous levodopa infusion (C), and 3) intermittent levodopa injection (I). Two weeks after the treatment, severity of LID was scored. Expression of mRNA on the operated side was compared to that on the unoperated side by real time RT-PCR.ResultsLID was observed in group I. Both D1 and D2 receptor mRNAs were elevated in group I, but only D2 receptor mRNA expression was elevated in group C. Enkephalin mRNA in group C and dynorphin mRNA in group I were elevated, indicating that the direct pathway is conversely dominant over the indirect pathway in LID. Adenosine A2A receptor mRNA was elevated and enkephalin mRNA was not elevated in group I.
Therefore, elevated A2A receptor may be in association with LID expression through inhibiting the indirect pathway.ConclusionContinuous or intermittent mode of levodopa administration affects dopaminergic and non-dopaminergic receptor expression. Elevated A2A receptor expression by intermittent levodopa administration may be related to LID pathogenesis.
Pe-055-3
Temporarily restricted expression of alpha-synuclein using Tet-off system to establish PD mice model
1Department of Neurology, Neurological Institute, Graduate school of Medical Sciences, Kyushu University，2Department of Neuropsychiatry, School of Medicine, Keio University
○Hiroo Yamaguchi1，Atsushi Fujita1，Hayato Une1，Yinan Zhao1， Kenji Tanaka2，Jun-ichi Kira1
Object: Various animal models, in which α-synuclein is overexpressed in DA neurons, have been tried for Parkinson’s disease (PD) animal model. For true animal models of PD，α-synuclein should be overexpressed in DA neurons not in the developmental stage but rather in the adult stage. Among several inducible transgene systems, Tet-off system is more efficacious for regulating gene expression in a temporally restrictive manner. To investigate the molecular mechanism of DA neuron death related to α-synuclein pathology and to establish PD mice model, we aimed to establish the mice, in which α-synuclein is overexpressed in DA neurons in a temporarily restrictive manner using Tet-off system.Methods: We used TetO-α -SynA53TTg/+ mice and DAT-tTATg/+ mice. TetOα -SynA53TTg/+mice express mutant A53T human α -synuclein (α -SynA53T) regulated by a tetracycline operator. DAT-tTATg/+mice express a tetracycline-controlled transactivator protein (tTA) driven by the mouse dopamine transporter (DAT) promoter. We mated TetOα -SynA53TTg/+mice with DAT-tTATg/+mice and analyzed their offspring including TetOα -SynA53TTg/+; DAT-tTATg/+, TetO-α -SynA53TTg/+; DAT-tTA+/+, TetO-α -SynA53T+/+; DAT-tTATg/+, and TetO-α-SynA53T+/+;DAT-tTA+/+mice.Results and conclusion: Immunohistochemical analysis showed that TetO-α-SynA53TTg/+;DAT-tTATg/+mice successfully expressed α-SynA53T in DA neurons in the SN in the absence of doxycycline (Tet-off). We will analyze these mice whether they show DA neurons loss. TetO-α-SynA53TTg/+;DAT-tTATg/+mice are useful for temporarily restricted expression of α-synuclein.
Pe-055-4
Can exercise modify Parkinson’s disease?-using a new mouse model of Parkinson’s
disease-Department of Neurology, Graduate School of Medicine, Kyoto University
○Shinya Okuda，Norihito Uemura，Ryosuke Takahashi
[Background and Purpose] It has been shown that exercise improve the cognitive dysfunction and memory loss. In terms of the molecular mechanisms, exercise affects the expression of brain-derived neurotrophic factor (BDNF) through the PGC-1α/FNDC5 pathway in the hippocampus. Several clinical data showed that exercise is also effective for Parkinson’s disease, but the molecular mechanisms are unknown. Our purpose of this research is to validate the disease modifying effect of exercise on Parkinson’s disease, using a new mouse model. [Methods]
First, to establish new disease model, we injected α-Synuclein (α-Syn) fibrils into the striatum of the α-Syn BAC (bacterial artificial chromosome) transgenic mouse that overexpress α-Syn. We’ll analyze the α-Syn-positive Lewy Body pathology and the number of dopaminergic neurons and evaluate their motor function one, three, and six months after injection. Further, we plan to force these model mice to exercise with running wheels and to evaluate whether their pathology and motor function can be improved or not. [Result] In preliminary experiments, injection of α-Syn fibrils caused the propagation of α-Syn-positive Lewy Body pathology. [Conclusion] We’ve confirmed that inoculation of α-Syn fibrils induce Lewy Body pathology. Hereafter, we begin to establish the new disease model.
Pe-055-5
Pathological role of GBA2 in GBA1-deficient neuronopathic Gaucher’s disease model of medaka
1Department of Neurology, Graduate School of Medicine, Kyoto University，
2Laboratory for Molecular Membrane Neuroscience, RIKEN Brain Science Institute，3Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University，4Department of radiation Genetics, Graduate School of Medicine, Kyoto University
○Etsuro Nakanishi1，Norihito Uemura1，Hisako Akiyama2， Masato Kinoshita3，Hodaka Yamakado1，Shunichi Takdeda4， Yoshio Hirabayashi2，Ryosuke Takahashi1
[Objective] Recent genetic studies have identified heterozygous mutations in the GBA1 gene as a strong risk factor for sporadic Parkinson’s disease (PD). The GBA1 mutations are responsible for Gaucher’s disease (GD). We have reported that GBA1 knock-out (KO) medaka can survive long enough for pathological analysis of disease progression in contrast to the perinatal death of GBA1 KO mice. This neuronopathic GD medaka model displays α-synuclein accumulation in the brains and has a potential to investigate the mechanisms of α-synuclein accumulation in GD and GBA1-related PD. The non-lysosomal β-Glucosidase (GBA2), which is localized at the endoplasmic reticulum and Golgi apparatus, also cleaves glucosylceramide to glucose and ceramide. A recent study has reported that the deletion of GBA2 rescues the visceral manifestation of type1 GD mice model. To date, the relationship between GBA1 and GBA2 in the central nervous system in GD and GBA1-related PD remains unclear. Our objective is to clarify the pathological role of GBA2 in the central nervous system in GD and GBA1-related PD. [Methods] We planned to generate GBA2 KO medaka by clustered regularly interspaced short palindromic repeat (CRISPR) / CRISPR-associated nuclease (Cas9) system. [Results] We have successfully generated GBA2 KO medaka by CRISPR / Cas9.
GBA2 KO medaka appear to show no developmental abnormalities compared to wild-type medaka. [Conclusions] We will cross GBA2 KO medaka with GBA1 KO medaka to examine the genetic interaction between GBA1 and GBA2 in GD and GBA1-related PD.
542
-21日
㈯ ポス ター
（英 語)
Pe-055-6
Oxidative stress and iron accumulation in the brain of the Zitter rat
1Department of Neurology, Dokkyo Medical University，2Department of Neuroimmunology, Medical University of Vienna，3Department of Histology and Neurobiology, Dokkyo Medical University
○Taro Kadoaki1,2,3，Hans Lassmann2，Shuichi Ueda3，Cornelia Schuh2， Koichi Hirata1
[Objective] Neurodegenerative diseases such as Parkinson’ s disease, multiple system atrophy and multiple sclerosis have much iron accumulation in the brain.
We investigated relationship between iron accumulation, inflammatory cells and oxidative stress in the brain of the zitter rat, which is one of neurodegenerative disease model. [Methods] From 7 to 9-month-old of Zitter rats (n=10) and age-matched Lewis rats were used. ED1 staining, CD3 staining, Turnbull’ s blue (TBB) and p22phox staining, were performed. We created an iron accumulation score from 0 to 3 in order to measure the degree of iron accumulation. The density of ED1-positive cells and CD3-positive cells were measured using ImageJ densitometry. And we counted the number of p22phox positive cells. [Results] Iron accumulation was prominent in the cerebellum and substantia nigra (SN) in all Zitter rats. Iron deposition score of Zitter rats was significantly higher than that of Lewis rats in the SN and cerebellum. ED1 was significantlly more in the cerebellum in the Zitter rat. There were no significant differences in the CD3 and p22phox cells. [Conclusions] Much iron deposition in the brain of the aging Zitter rat did not yield oxidative stress. It might be gone in young period.
Pe-055-7
Neurotoxic roles of astrocytic gap junctional protein connexins 30 in MPTP toxicity
Department of Neurology, Neurological Institute, Graduate school of Medical Sciences, Kyushu University
○Atsushi Fujita，Hiroo Yamaguchi，Hayato Une，Yinan Zhao，
Jun-ichi Kira
Aim: Connexin (Cx) 30 is a major gap junctional protein in astrocytes. Gap junctional intercellular communication (GJIC) plays an important role in pathogenesis of various neurological diseases. The association of GJIC with the pathogenesis of Parkinson’s disease has been reported using various animal models. MPTP is a widely used toxic agent for dopaminergic neurons mainly by inhibiting mitochondrial complex I.
Previous studies have shown that conversion from MPTP to MPP+ in astrocytes and transportation of MPP+ from astrocytes to neurons is an essential step for neural damage in acute stage, which is followed by astrogliosis in the striatum and substantia nigra along with dopaminergic neurons loss in chronic stage. We aimed to clarify the roles of Cx30 in MPTP toxicity using Cx30 knockout mice.Methods: MPTP (30 mg/kg) or saline was administered i.p to wild type and Cx30 knockout mice every 24 hours for 5 consecutive days. At 6 hours after the last injection, these mice were killed for analyses. Immunohistochemical and biochemical analyses were performed to compare glial reaction and dopaminergic neuron damage between the groups.Results:
Cx30 knockout mice showed less bradykinesia and dopaminergic neuron loss together with milder astrocytic reaction (less GFAP and Cx43 expression), as compared with wild type mice.Discussion: It is assumed that in natural state various toxic substances from damaged or dead neurons spread via gap junction as "death signaling." The hampered signaling by knockout of gap junctional protein might produce the protective effect in MPTP toxicity
Pe-056-1
Mutations of a mitochondrial protein CHCHD2 lead to Parkinson’s disease-like phenotypes
1Research institute for diseases of old age, Juntendo University Graduate School of Medicine，2Department of Neurology, Juntendo University Graduate School of Medicine，3Department of Research for Parkinson’ s Disease, Juntendo University Graduate School of Medicine
○Hongrui Meng1，Chikara Yamashita2，Kahori Shiba2，Yuzuru Imai2,3， Hattori Nobutaka1,2,3
[Object] Mitochondrial dysfunction has been implicated in the etiology of Parkinson’s disease (PD), the second most common movement disorder characterized by the loss of dopaminergic neurons.
Mutations in the CHCHD2 gene, which code a mitochondrial inner membrane protein are linked to autosomal dominant form of familial PD. However, it is unclear how mutations in CHCHD2 produce neurodegeneration in familial cases of PD. In order to understand the physiological and pathological roles of CHCHD2, we produced transgenic Drosophila fly lines to model PD. [Methods] We produced transgenic fly lines that loss function of CHCHD2 and by using tissue- and cell-type-specific transcriptional system to express normal and T61I or R145Q of CHCHD2 mutants in neuronal and non-neuronal tissues. A variety of behavioral phenotypes associated with PD, viability and morphology appearances were observed. [Results] The deficiency of CHCHD2 in flies show mitochondrial and neuronal phenotypes associated with PD pathology, which include increased sensitivity to oxidative stress and loss of dopaminergic neurons with age. Furthermore, down regulation of CHCHD2 expression causes mitochondrial cristae defects and decreases ATP synthesis, these phenotypes are rescued by expression of hCHCHD2, but not T61I and R145Q PD mutants. [Conclusion] Our study suggests that the etiology of PD linked to CHCHD2 might contain both gain- and loss-of-function aspects. The Drosophila model thus recapitulates the essential features of the human disorder, and makes possible a powerful genetic approach to PD.
Pe-056-2
PD-linked mutation in DNAJC13 causes specific trafficking defect in endosomal pathway
1Division of Neurology, Department of Neuroscience & Sensory Organs, Tohoku University Graduate School of Medicine，2Department of Neurology, Sendai-Nishitaga Hospital
○Shun Yoshida1，Takafumi Hasegawa1，Ryuji Oshima1，
Junpei Kobayashi1，Naoto Sugeno1，Akio Kikuchi1，Atsushi Takeda2， Masashi Aoki1
Background: Recently, a missense mutation (p.N855S) in DNAJC13 gene has been identified in patients with familial forms of PD. DNAJC13 is associated with endosomal membrane.Objective: The aim of this study is to investigate the effect of mutant DNAJC13 on the vesicle transport machinery using cultured cellular model.Methods: COS7 cells were transfected with DNAJC13 together with Rab GTPase constructs. The expression and subcellular localization of DNAJC13 were examined using laser scanning microscope and Western blot analyses. To determine how the DNAJC13 mutant affects on the different endosomal pathway, cells over-expressing wt or mutant DNAJC13 were incubated in the culture media containing reference molecules including EGF and transferrin, and time-lapse images were acquired. To assess the subcellular distribution of α-synuclein in the presence of mutant DNAJC13, cells expressing α-synuclein were co-transfected either with wt or mutant DNAJC13 under the existence of cycloheximide and subjected to subcellular fractionation analysis.Result: While wt-DNAJC13 was co-localized with rab5A-positive endosome, N855S mutant was co-localized with not only rab5A but also Rab7 or Rab11A-positive endosome. The cargo-trafficking from early endosome to late endosome and/or recycling endosome was impaired in cells over-expressing N855S DNAJC13.
Moreover, endosomal retention of α -synuclein was observed in the cells expressing mutant DNAJC13.Conclusion: DNAJC13 mutation impairs endosomal cargo trafficking including α-synuclein and would contribute to the pathogenesis of the disease.
Pe-056-3
Parkin is involved in miR 132 processing
1Department of Neurology, Tokushima National Hospital，2Department of Clinical Reserch, Tokushima National Hospital
○Takao Mitsui1,2，Yukiko Maki2，Miki Fujimoto2，Masako Sogo2， Kayo Nagahama2
IntroductionParkin is known to be a multifunctional protein. We previously reported that parkin was preferentially localized in mitochondria and enhanced mitochondrial.
On the other hand, Parkin reportedly interacts with p53, a representative oncogene, which promotes post-trancriptional modification of several kinds of miRNA. In this study, we identified miRNAs associated with Parkin and examined involvement of Parkin for the processing of miRNA in mitochondria and the nucleus.Materials and MethodsUsing RD cells that stably expressing His-Parkin, UV cross-linking and immunoprecipitation (CLIP) were carried out. We cloned miRNA which bound to His-Parkin in nuclear and mitochondrial fraction. Furthermore, the expression was assayed using real-time PCR in obtained miRNA and pri-miRNA. We examined the expression of mature miRNA and pre-miRNA in primary fibroblasts obtained from PARK2 patients and normal controls.ResultsThe Drosha protein was detected in not only the nuclear fraction but also the mitochondrial fraction. Furthermore, Drosha which bound to Parkin was detected in both fraction. We identified miR132, miR638, miR26a-2 by cloning of miRNA. In Particular, miR132 and pri-miR132 were detected in mitochondria and nuclei. Mature miR132 and pre-miR132 were significantly decreased in fibroblasts from 4 PARK2 patients compared to those from controls.ConclusionsParkin seems to be involved in the processing of miR132 in the mitochondria and nuclei and the reduction of miR132 may play a pathophysiological role in neuronal dysfunction of PARK2.
Pe-056-4
Regulation of Parkin/ChPF expression
1Department of Clinical Research, Tokushima National Hospital，2Department of Neurology, Tokushima National Hospital
○Yukiko Maki1，Takao Mitsui1，Miki Fujimoto1，Kayo Nagahama1， Ryuji Kaji2
[Background]We reported that Parkin was located to intracellular mitochondria and promoted mitochondrial biogenesis. Parkin bound to Klokin 1, which carried it to mitochondria. Klokin 1 was a splicing variant of Chondroitin Polymerizing Factor (ChPF). ChPFΔ996 was detected as a different variant. We examined the association of Parkin with ChPF family and the effect of ChPF family on transcription of parkin gene.[Methods]Halo-ChPF family and His parkin were transfected to cultured cells.
The association was examined by coimmunoprecipitation. Also, we transfected ChPF family in cultured cells. The localization in cells of the endogenous Parkin was examined and parkin mRNA was assayed by real-time PCR. In addition, we constructed the reporter vector containing parkin promoter domain for a luciferase assay.[Results]In the coimmunoprecipitation assay, ChPF, ChPFΔ996, and Klokin1 were associated with parkin. The intracellular locatization of Parkin completely accorded with that of Klokin 1, corresponding to the mitochondria. On the other hand, ChPF and ChPF Δ996 were found inside and outside the mitochondria. In the cells with overexpression of Klokin 1, ChPF and ChPF Δ 996, the expression of the endogenous Parkin increased. In these cells, Parkin mRNA increased and transcription activity of the parkin gene also increased. When parkin was overexpressed, ChPF protein and its mRNA were not increased.[Conclusions]ChPF family promoted the Parkin transcription. But Parkin did not enhanced that of ChPF family.