成熟ラット上衣下脳組織由来神経系幹細胞からのドパミン産生神経細胞の誘導

金沢大学医薬保健研究域医学系昨年度は,成熟ラット上衣下脳組織由来の細胞を培養し,免疫組織化学染色により,それらの細胞の約90%がnestin 陽性で神経系幹細胞であることが確認された.平成13年度は,この神経系幹細胞からドパミン産生神経細胞を誘導する実験を行った.1)ドパミン産生能の評価:前年度の実験で得られた細胞を培養し,培養液中に遊離されるDOPAおよびdopamineの量をHELC-PC法で測定したが,その値は測定限界以下であった.しかしながら,本実験で得られた神経系幹細胞が脳への移植により,生着可能かどうかを確認するため以下の実験を行った.2)lacZ遺伝子の導入:ドパミン産生能を獲得した培養神経細胞に,lacZ遺伝子を組み込んだアデノウイルスベクターを感染させ,移植細胞とした.3)細胞移植:lacZ遺伝子を導入された培養神経細胞を1×10^4に調節し,定位脳装置を用いてParkinson病モデルラットの線条体に移植した.4)組織学的検索:移植後4週のラットを用いた.・移植細胞の生存の確認:β-galactosidaseに対する免疫化学染色では一部細胞が陽性であり,移植細胞が脳内で生存していた.・ドパミン産生能の確認:tyrosine hydroxylase(TH)に対する免疫化学染色を行った.宿主側に元々存在するTH陽性細胞との識別を行うため,β-galactosidaseとの2重染色を行ったが,両者が陽性の細胞はほとんどみられなかった.つまり,今回の実験では神経系幹細胞は移植により脳内で生存可能であることは確認されたが,特異的にドパミンを産生する細胞への分化は確認できなかった.研究課題/領域番号:12770746, 研究期間(年度):2000-2001出典：「成熟ラット上衣下脳組織由来神経系幹細胞からのドパミン産生神経細胞の誘導」研究成果報告書　課題番号 12770746（KAKEN：科学研究費助成事業データベース（国立情報学研究所））（https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-12770746/)を加工して作

感覚伝導路の形成・再生における細胞接着分子および細胞外基質の機能に関する研究

金沢大学附属病院新生マウス(0日齢)および幼若マウス(7,14,28日齢)をパラフォルムアルデヒドにて経心臓的に潅流固定を行った後,脊髄後根,神経節,後根脊髄を一塊として摘出した.取り出した組織をOTC compoundに包埋し液体窒素で凍結して,厚さ6μmの凍結切片を作成した.この切片上で,E-cadherin,L1,NCAMおよびagrinの発現を免疫組織化学的手法により確認した.NCAM,L1は光顕的観察では,脊髄後根神経節,後根,脊髄上でびまん性に陽性であり,免疫電顕での観察では,ほとんどすべての無髄神経の軸索膜に陽性であった.E-cadherinは,光顕的観察で後根神経節の一部,脊髄後角の第II層で陽性であり,免疫電顕での観察では,無髄神経線維の一部(約20%)に陽性であった.これまでの研究から,E-cadherinは,一部の無髄感覚神経線維に陽性であることが確認された.発生段階におけるE-cadherinの発現様式は,ある特定の知覚伝導を司る感覚神経線維にのみ特異的に発現していると考えられ,痛覚伝導路形成における軸索束形成に重要な役割を果たしていると考えられた.また,agrinに関しては,光顕的観察において脊髄後角内でわずかに陽性であった.Agrinはneuromuscular junctionにおけるacetylcholine receptorの集簇を司る細胞外基質である.現時点において,免疫電顕の手法を用いた場合,抗体の浸透性および細胞外基質の固定性の問題から,光顕的鑑札でみられた脊髄後角でのagrin陽性の所見が,どの種類の細胞由来かは確認できなかった.今後は潅流固定法,切片作成法,抗体濃度および抗原の賦活法を改善し,痛覚伝導路形成におけるagrinの役割を解析する予定である.研究課題/領域番号:10770675, 研究期間(年度):1998 – 1999出典：「感覚伝導路の形成・再生における細胞接着分子および細胞外基質の機能に関する研究」研究成果報告書　課題番号10770675（KAKEN：科学研究費助成事業データベース（国立情報学研究所））（https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-10770675/)を加工して作

マウス知覚神経における接着分子(上皮型カドヘリン)の発現: 特に軸索束形成における役割について

取得学位 : 博士（医学）, 学位授与番号 : 医博甲第1081号, 学位授与年月日：平成5年3月25日,学位授与年：199

Structures of SMG1-UPFs Complexes: SMG1 Contributes to Regulate UPF2-Dependent Activation of UPF1 in NMD

SummarySMG1, a PI3K-related kinase, plays a critical role in nonsense-mediated mRNA decay (NMD) in mammals. SMG1-mediated phosphorylation of the UPF1 helicase is an essential step during NMD initiation. Both SMG1 and UPF1 are presumably activated by UPF2, but this regulation is incompletely understood. Here we reveal that SMG1C (a complex containing SMG1, SMG8, and SMG9) contributes to regulate NMD by recruiting UPF1 and UPF2 to distinct sites in the vicinity of the kinase domain. UPF2 binds SMG1 in an UPF1-independent manner in vivo, and the SMG1C-UPF2 structure shows UPF2 recognizes the FRB domain, a region that regulates the related mTOR kinase. The molecular architectures of several SMG1C-UPFs complexes, obtained by combining electron microscopy with in vivo and in vitro interaction analyses, competition experiments, and mutations, suggest that UPF2 can be transferred to UPF1 within SMG1C, inducing UPF2-dependent conformational changes required to activate UPF1 within an SMG1C-UPF1-UPF2 complex

Prediction of carotid artery in-stentrestenosis by quantitative assessment ofvulnerable plaque using computed tomography

Background and purpose To assess the relationship between plaque volume evaluated by multidetector computed tomographic angiography (MDCT) and in-stent restenosis (ISR) after carotid artery stenting (CAS). Materials and methods From a retrospectively maintained database, data were collected for 52 patients with carotid artery stenosis treated with CAS between 2007 and 2012. We defined ISR of ≥ 50% as a peak systolic velocity ≥ 200 cm/s on echo-duplex scan. Carotid plaques were subdivided into four components according to radiodensity in Hounsfield units (HU) as follows: 600 HU. Risk factors that influenced ISR were compared using univariate and multivariate Cox regression analyses. Results During a median follow-up period of 36 months, ISR of ≥ 50% was detected in five patients (9.6%). In the univariate Cox proportional hazard regression analysis, renal insufficiency, coronary artery disease, total plaque volume, and plaque volumes with radiodensities < 0 and ≥ 600 HU increased the risk for ISR (P < 0.10). When the significant risk factors determined from the univariate analysis were subjected to a multivariate analysis, only the volumes of the plaque components with radiodensities < 0 HU independently predicted the development of ISR (hazard ratio: 1.041; 95% confidence interval: 1.006–1.078; P = 0.021). Conclusion Our data suggest that the high volume of the plaque components with radiodensities < 0 HU was independently associated with the increased risk of ISR after CAS. Quantitative and qualitative tissue characterizations of carotid plaques using MDCT might be a useful predictive tool of the development of ISR.Embargo Period 12 month

Unique Venous Drainage of a Sphenoid Wing Dural Arteriovenous Fistula with Ocular Symptoms

Background: Dural arteriovenous fistulas (DAVFs) presenting with ocular symptoms, such as exophthalmos and chemosis, are commonly situated in the cavernous sinus (CS). DAVFs at the sphenoid wing with a drainage route into the superior orbital vein (SOV) should be considered as one of the differential diagnoses of ocular symptoms. Case Description: A 41-year-old woman presented with progressive left-sided chemosis and proptosis after left pulsating tinnitus that disappeared spontaneously. Cerebral angiography showed that the fistula was situated along the inferior edge of the superior orbital fissure on the greater sphenoid wing and drained solely into the SOV without flowing into the CS that caused ocular symptoms. Transvenous selective catheterization was performed via the facial vein and SOV. The fistula was then embolized using detachable coils. Conclusions: After embolization, the ocular symptoms resolved, and the patient was discharged without neurologic deficit. Herein, we discuss the developmental mechanism of the unique drainage pattern, including the clinical symptoms and anatomic features of greater sphenoid wing DAVFs. © 2016 Elsevier Inc.Embargo Period 12 month

Intraparenchymal pneumocephalus caused by ethmoid sinus osteoma

金沢大学医薬保健研究域医学系We report a 57-year-old man with intraparenchymal pneumocephalus caused by ethmoid sinus osteoma. He had a history of severe allergic rhinitis, which caused him to frequently blow his nose, and he was referred to our hospital with headache and mild left hemiparesis. CT scans revealed a large volume of intraparenchymal air entrapped in the right frontal lobe related to an osteoma in the ethmoid sinus. The osteoma eroded the upper wall of the sinus and extended into the anterior cranial fossa. At operation, we observed that the osteoma had protruded intracranially through the skull base, disrupted the dura and extended into the frontal lobe. To our knowledge, this is the first report of a patient with intraparenchymal pneumocephalus caused by an ethmoid sinus osteoma. © 2009 Elsevier Ltd. All rights reserved

Long-term predictive factors of the morphology based outcome in bare platinum coiled intracranial aneurysms: Evaluation by pre- and post-contrast 3D time-of-flight MR angiography

Purpose Our aim was to identify long-term predictive factors of the morphology-based outcome (MBO) of bare platinum coiled intracranial aneurysms. Materials and Methods A retrospective analysis of 96 bare platinum coiled intracranial aneurysms followed up from 1997 to 2016 using pre- and post-contrast 3D time-of-flight MR angiography (MRA) was performed. Logistic regression analysis was used to identify factors associated with a positive history of surrounding coil mass enhancement (SCME) and poor MBO. Spearman's rank correlation test was used to analyze the relationship between the initial angiographic result (IAR) class, sequential change of the SCME category, and MBO grade. Results Factors independently associated with poor MBO were incomplete IAR (OR=14.94, 95%CI: 2.46, 289.21, P=0.002) and a history of SCME (OR=4.13, 95% CI: 1.05, 18.65, P=0.043). The MBO grade strongly correlated with the IAR class (correlation coefficient [r]=0.84, P&lt;0.0001). MBO grade correlated with sequential change of the SCME category (r=0.56, P&lt;0.0001). The sequential change of the SCME category correlated with IAR class (r=0.53, P&lt;0.0001). Conclusion Although IAR and its class were strong long-term predictive factors of MBO, a history of SCME and upgrading of sequential change of SCME category were also long-term predictive factors of the MBO of bare platinum coiled intracranial aneurysms

Aneurysm clipping after partial endovascular embolization for ruptured cerebral aneurysms

The aim of this study was to investigate the advantages and disadvantages of a two-stage treatment for ruptured cerebral aneurysms; partial embolization in acute stage followed by clipping in chronic stage of subarachnoid hemorrhage. Between April 1997 and August 1999, twenty ruptured cerebral aneurysms were initially treated endovasculary using Guglielmi detachable coils in our institution. Among them, complete embolization could not be achieved in 6 lesions. For these lesions, subsequent clipping was added. The radiological and operative findings, and outcomes of these cases were retrospectively reviewed. In 1 case, rerupture occurred during the endovascular procedure. Rerupture was not observed in any cases in the postembolization period. In 2 cases, complications related to the clipping but not the endovascular procedure occurred. These complications included impaired visual acuity for unverified reasons, and memory disturbance due to sacrifice of a perforator arising from the anterior communicating artery. In 3 cases, coil extraction was needed during the clipping, because the loops of the coil extended into the residual neck. Complications related to coil extraction were not observed in these 3 cases. Acute partial embolization of ruptured aneurysm appears to be effective for the prevention of subsequent rerupture during the subacute period, in which treatment for vasospasm should be performed, and the clipping procedure. However, in the case of relatively large aneurysms, small arteries or other normal structures behind the aneurysm cannot be observed directly during surgery, because of the immovability of the embolized aneurysm. Further, complete clip closure is impossible when loops of coil herniate into the neck. In such situations, coil extraction with or without resection of the aneurysm might be necessary, and care must be taken not to damage parent artery and surrounding vessels

Significance of volume embolization ratio as a predictor of recanalization on endovascular treatment of cerebral aneurysms with Guglielmi detachable coils

The purposes of this study are, firstly, to define the relationship between volume embolization ratio (VER) and degree of angiographical occlusion in endovascular treatment with Guglielmi detachable coils, and secondly, to examine influences of neck and dome sizes of aneurysms on the VER and the angiographical treatment result, and thirdly, to determine the relationship between the VER and the recanalization of coiled aneurysms. Fifty-two aneurysms in 46 patients were examined. VER ranged 8.1-31.9% (mean 18.5%). The mean VERs of each categories based on angiographical treatment results were 23.1% in complete occlusion, 16.1% in neck remnant and 12.2% in incomplete occlusion, respectively. The VER correlated significantly with both neck and dome size, while the angiographical treatment result was only affected by neck size. Five aneurysms showed aneurysmal recanalization among followed-up 41 aneurysms. All recanalized aneurysms were large, and their VERs were in range of 10.4-17.6%. Measurement of VER is useful to estimate the degree of occlusion objectively and to predict the aneurysmal recanalization. A small aneurysms with a small neck is relatively easy to achieve high VER and angiographical complete occlusion, with the consequence of less recanalization. On the other hand, a large aneurysm is liable to recanalize due to low VER, even if there was little filling of contrast medium in the aneurysmal cavity