Ehime University Institutional Repository

Ehime University (愛媛大学)博士(工学)doctoral thesi

ZULKIFLI, TAHIR

English

The Study on Automated HTML5 Offline Services to Overcome Low Reliability of Network Connectivity for Web-based Decision Support System Applications

中曽, 久雄

Japanese

アメリカ憲法における尊厳概念の展開とその意義(1)

ロックウッド・カレン,

小田, 敬美[訳]

「経験的に」教える経験を積む : いかにして教員はロースクールを法実務につなぐのか?

Ehime University (愛媛大学)博士(農学)thesi

長谷川, 雄基

HASEGAWA, Yuki

 
 
（第５号様式）  
                        学位論文審査の結果の要旨 
 
    氏    名 長谷川 雄基 
審 査 委 員 
       
 
          
論  文  名 
 
摩耗が生じた農業用水路コンクリートの劣化診断手法に関する研究 
 
審査結果の要旨 
 
農業水利施設の維持管理を的確に遂行するためには、施設の現況性能の正確な診断と性能低下に
つながる変状特性の定量的な評価が不可欠である。農業用コンクリート水路の代表的な変状として、
ひび割れと骨材露出がある。比較的小規模な水路を対象として、ひび割れの挙動特性を調査したと
ころ、底版による拘束のみの無筋水路においては、ひび割れ幅が年間に 1mm 程度変動する場合も
あることが分かった。無筋水路と RC 水路において、ひび割れ幅の季別の日変動量と年変動量を比
較すると、RC 水路は水路躯体と地盤との拘束や躯体内の鉄筋が気温変動による収縮・膨張に対し
て拘束効果を発揮した。一方、無筋水路では、ひび割れ幅の変動を抑制する要因が水路躯体と地盤
との拘束条件のみであることから、ひび割れ幅の変動量は無筋水路の方が大幅に大きくなることが
明らかとなった。 
一方、骨材露出の原因となる摩耗は、すべての環境下で普遍的に進行する劣化である。現行の水
利施設の維持管理に係るマニュアル・書籍類では、摩耗のメカニズム、供用年数の経過と摩耗の進
行との関係や摩耗が要求性能に及ぼす影響について、概念的には示されているものの、劣化の定量
的な評価指標や診断手法の確立には至っていないのが現状である。 
本研究は、農業用水路コンクリートの摩耗に対する劣化診断手法について検討したものである。
まず現地調査を通して、長期供用された実水路コンクリートの摩耗特性を評価した。続いて、実水
路の調査結果から得られた課題の解決を目的として、詳細な摩耗の進行過程と粗骨材の脱落条件な
らびに劣化診断を行う際の評価指標について、実験的に検証した。さらに、複合劣化に対する診断
手法の一例として、摩耗が生じた農業用水路コンクリートにおける中性化の評価手法について検討
した。最後に、本研究で得られた成果を活用し、摩耗に対する劣化診断手法の具体的な方法および
手順を提案した。 
実水路の調査結果として、同一水路の側壁と底版とでは、側壁の方が表面粗さは大きいことが確
認できた。側壁と底版とでは摩耗のメカニズム自体が異なり、側壁では流水に起因したエロージョ
ン作用および溶脱作用が卓越し、底版では堆積した砂礫によるすり磨き作用と転がり摩擦作用が卓
 
 
越すると考えられた。また、摩耗が生じた水路コンクリートの劣化診断を行う際には、取得できる
データのばらつきが比較的小さい側壁の水中部では、測定位置を任意に設定することが有効である
と考えられた。本調査対象水路においては、表面粗さの数値は側壁の方が大きかったものの、劣化
の進行程度は粗骨材が部分的に脱落している底版の方が進行していることが確認された。よって、
水中部という同一の環境であっても、部材位置ごとに劣化診断を行う必要があると考えられた。 
室内実験で得られた結果として、摩耗が生じた水路コンクリート壁面の劣化診断および劣化予測
を行う上で、粗骨材の脱落開始時期を推定できる実験式を示した。また、粗骨材の露出により表面
粗さが増加した供試体の粗度係数を推定したところ、粗度係数は設計時に使用される基準値と同程
度であった。このことから、摩耗に伴う水路コンクリートの粗骨材の露出は、必ずしも通水性能の
低下要因にはなり得ないと推察された。実水路において比較的簡便に取得可能な表面粗さから摩耗
深さを推定する手法を検討したところ、表面粗さ指標と摩耗深さの指標は、すべて継時的に同一の
傾向で増加することが確認できた。それぞれの指標同士の関係を分析し、実水路の表面粗さを計測
することで、間接的に摩耗深さを推定できる実験式を示した。 
複合劣化に対する診断手法の一例として、摩耗が生じた農業用水路コンクリートにおける中性化
の診断手法について検討したところ、水路の水中部における中性化速度係数と表面粗さ指標の一つ
である表面積比との間には、非常に強い負の相関関係のあることが確認できた。この理由として、
表面粗さが大きい水路ほど、コンクリート表面からは中性化が進行したモルタルが多く消失し、見
かけの中性化深さが小さくなったためと考えられた。一方、表面粗さが小さい水路においては、コ
ンクリート表面近傍における pH の低下が生じたモルタル部分がコンクリートの表面に多く残存し
ていたため、中性化深さが大きくなったと推察された。 
以上の研究成果を活用し、実水路コンクリートの摩耗に対す劣化診断のフローを示した。対象と
する劣化の進行状態は粗骨材の露出開始以降であり、評価項目は、通水性能に関わる粗度係数、構
造耐力に関わる断面の消失量、他の劣化の進行評価に必要な摩耗深さである。本研究から得られた
実験式と既往の研究で示された実験式の両者を組み合わせることで、原位置において比較的簡便に
取得可能な表面粗さの計測結果のみから、上記の項目をそれぞれ評価することが可能な手法を提案
した。 
 
本論文の学位論文公開審査会は、平成 28 年 2 月 6 日に愛媛大学農学部において開催され、論文
発表と公開審査が行われた。引き続いて、学位論文審査会が開かれ、論文の内容について審査した
結果、審査員全員一致して、本論文は博士（農学）の学位を授与するに値すると判定した。 


摩耗が生じた農業用水路コンクリートの劣化診断手法に関する研究

真鍋, 邦男

MANABE, Kunio

（様式５）(Style5) 
学位論文全文に代わる要約 
Extended Summary in Lieu of Dissertation 
 
 
氏名： 
Name 
真鍋 邦男 
 
学位論文題目： 
Title of Dissertation 
出芽酵母Saccharomyces cerevisiae液胞塩基性アミノ酸輸送系に関する研究 
 
 
学位論文要約： 
Dissertation Summary 
出芽酵母（Saccharomyces cerevisiae）の液胞は能動的アミノ酸輸送により細胞全体の約70〜90 %の
塩基性アミノ酸（リジン／ヒスチジン／アルギニン）を蓄積する[1, 2]。その分子機構についてMajor 
Facilitator Superfamily中のVBAサブファミリーに属するVba1p, Vba2p, Vba3pが液胞への塩基性アミ
ノ酸取り込みに機能し[3]、Amino Acid/Auxin Permease Superfamilyに属するAvt1pが液胞プロトン/ヒ
スチジンアンチポーターとして機能することが報告されている[4, 5]。しかし、VBA及びAVT多重破
壊株由来の液胞膜小胞でも依然として各塩基性アミノ酸の取り込み活性が検出されたことから、未
知のトランスポーターの存在が示唆されていた。最近、Transporter-Opsin-G protein coupled receptor 
(TOG) Superfamilyに属するラットのPQLC2及び線虫のLAAT-1がリソソームの塩基性アミノ酸輸送
に関与することが報告された[6, 7]。これらはプロリン-グルタミン配列を含むPQループとよばれる保
存配列を2カ所有する7回膜貫通型タンパク質である[8]。本研究では、出芽酵母のTOG Superfamilyに
属する液胞膜タンパク質Ypq1p、Ypq2p、Ypq3pの塩基性アミノ酸輸送能を単離液胞膜小胞のATP依
存的な14C標識アミノ酸輸送活性測定により検討した。 
 Ohsumi ら[9]の方法に従って単離した液胞膜小胞は YPQ1、YPQ2、YPQ3 遺伝子のいずれを破壊し
ても ATP 依存的なチロシンの取り込み活性に大きな変化はなかったが、リジンの取り込み活性が
YPQ1遺伝子破壊により大幅に低下した（Fig. 1A, D）。液胞膜小胞のV-ATPaseレベルや小胞内のATP
依存的な酸性化はYPQ1遺伝子破壊によってほとんど変化せず（Fig. 2A, B）、YPQ1を過剰発現する
と液胞膜小胞のリジン取り込みが大幅に増加し、この活性はCCCP添加により大きく阻害されたこと
から（data not shown）、Ypq1pはプロトン濃度勾配を駆動力とするリジンの取り込みに特異的に機能
することが示唆された。 
 
 
（様式５）(Style5) 
 
  
液胞膜小胞のアルギニンの取り込み活性はYPQ1とYPQ2各遺伝子破壊により部分的に低下し、
YPQ1とYPQ2の二重破壊によってさらに低下した。一方、YPQ3遺伝子破壊ではアルギニン取り込み
活性に大きな変化は見られなかった（Fig. 1B）。液胞膜小胞のアルギニン取り込みについてはアル
ギニン/ヒスチジン交換輸送活性が報告されており[10]、この性質に注目した。野生株及びypq1∆株か
ら単離した液胞膜小胞のATP依存的なアルギニン取り込みはヒスチジン添加によって増加したのに
対し、ypq2∆株由来の小胞ではほとんど増加しなかった（Fig. 3）。また、ヒスチジンを前負荷した
野生株由来の小胞は、 
 
希釈により小胞内外でヒスチジン濃度勾配を形成させるとATP非存在下でもアルギニンを取り込ん
だが、YPQ2遺伝子を破壊するとこの取り込みが大きく抑えられ（Fig. 4A）、異種プロモーターより
YPQ2を発現させると、発現量に依存して取り込み活性が変化した（Fig. 4B）。以上の結果は、Ypq2p
がアルギニン/ヒスチジン交換輸送の本体であることを示唆している。さらに、14C標識ヒスチジンを
前負荷した小胞への未標識アルギニン添加によって排出されたヒスチジン量の測定より交換輸送の
アルギニンとヒスチジンの量比は約1:1であることが示された（data not shown）。 
（様式５）(Style5) 
 
 
液胞膜小胞のヒスチジン取り込み活性は YPQ3遺伝子破壊により部分的に低下した一方、YPQ1及
びYPQ2遺伝子の破壊によってはほとんど変化しなかった（Fig. 1C）。ypq1∆ypq2∆ypq3∆avt1∆株から
単離した液胞膜小胞は各塩基性アミノ酸（リジン、アルギニン、ヒスチジン）の取り込み活性がほぼ
消失したが、この株に YPQ3をGPDプロモーターにより過剰発現すると小胞のヒスチジン取り込み
のみが検出された（Fig. 5）。このYpq3p依存的なヒスチジン取り込みは未標識ヒスチジンの添加によ
って顕著に阻害されたが、その他の未標識アミノ酸の添加ではほとんど阻害されなかった。また、
Concanamycin A、CCCP、Nigericin添加によっても大幅に阻害された（Fig. 6A, B）。以上の結果から、
Ypq3pがプロトン濃度勾配に依存してヒスチジンを特異的に液胞内へと取り込み、Avt1pと重複して
機能することが示唆された。また、ネイティブプロモーターにより発現したYpq3p-HA3の細胞内レベ
ルはヒスチジン飢餓によって増加した一方、リジンもしくはアルギニンの飢餓では大きく変化しなか
ったことから、Ypq3pはその輸送基質によって特異的な発現制御を受けることが示唆された（Fig. 7）。 
 
さらにアルギニン/ヒスチジン交換輸送の駆動力となるヒスチジン取り込みへのAvt1pとYpq3pの
関与について検討したところ、ypq3∆株由来小胞のATP依存的なアルギニン取り込みはヒスチジン添
加によって野生株由来の小胞と同様に増加したが、avt1∆株由来の小胞ではypq2∆株由来の小胞同様
増加しなかった（Fig. 3）。その一方で、avt1∆株由来の小胞はヒスチジン濃度勾配を形成させると野
生株由来の小胞と同様にアルギニンを取り込んだ（Fig. 4A）。したがって、Ypq2pによるアルギニ
ン/ヒスチジン交換輸送の駆動力となるヒスチジン濃度勾配はin vitroではAvt1pによって形成される
ことが示唆された。 
（様式５）(Style5) 
 
 
 
 本研究では、液胞膜タンパク質である Ypq1p 及び Ypq3p がプロトン濃度勾配を利用してリジン・
ヒスチジンの液胞内への取り込みにそれぞれ機能することが示唆された。一方、Ypq2pは長年実体が
不明であったアルギニン/ヒスチジン交換輸送体として機能し、その駆動力となるヒスチジン濃度勾
配の形成にAvt1pが機能することが明らかとなった。ypq1∆ypq2∆ypq3∆avt1∆株から単離した液胞膜小
胞において塩基性アミノ酸の取り込みがほぼ消失したことから、in vitro では Ypq タンパク質及び
Avt1pは主要な塩基性アミノ酸取り込み系として機能すると考えられる。最近、Ypq1pがリジン飢餓
条件でユビキチン化され、液胞内で分解されることが報告された[11]。本研究においても、Ypq3p が
ヒスチジン飢餓により細胞内レベルが増加することを新たに見出し、Ypqタンパク質の合成と分解の
調節が細胞内外のアミノ酸濃度の変化に応答して細胞の環境適応に機能する可能性が示唆された。
Ypq タンパク質による基質輸送分子機構のさらなる解析と細胞内レベルの調節機構の解明はその生
理機能の理解につながるとともに、液胞内に塩基性アミノ酸を高度に蓄積する意義についても重要な
情報を提供することが期待される。 
（様式５）(Style5) 
【参考文献】 
 
[1] Wiemken A, and Durr M. (1974). Characterization of amino acid pools in the vacuolar compartment 
of Saccharomyces cerevisiae. Arch. Microbiol. 101, 45-57. 
 
[2] Kitamoto K, Yoshizawa K, Ohsumi Y, and Anraku Y. (1988). Dynamic aspects of vacuolar and 
cytosolic amino acid pools of Saccharomyces cerevisiae. J. Bacteriol. 170, 2683-2686. 
 
[3] Shimazu M, Sekito T, Akiyama K, Ohsumi Y, and Kakinuma Y. (2005). A family of basic amino acid 
transporters of the vacuolar membrane from Saccharomyces cerevisiae. J. Biol. Chem. 280, 
4851-4857. 
 
[4] Russnak R, Konczal D, and McIntire S. L. (2001). A family of yeast proteins mediating bidirectional 
vacuolar amino acid transport. J. Biol. Chem. 276, 23849-23857. 
 
[5] Tone J, Yoshimura A, Manabe K, Murao N, Sekito T, Kawano-Kawada M, and Kakinuma Y. (2015). 
Characterization of Avt1p as a vacuolar proton/amino acid antiporter in Saccharomyces cerevisiae. 
Biosci. Biotechnol. Biochem. 79, 782-789. 
 
[6] Liu B, Du H, Rutkowski R, Gartner A, and Wang X. (2012). LAAT-1 is the lysosomal lysine/arginine 
transporter that maintains amino acid homeostasis. Science. 337, 351-354 
 
[7] Jézégou A, Llinares E, Anne C, Kieffer-Jaquinod S, O'Regan S, Aupetit J, Chabli A, Sagné C, 
Debacker C, Chadefaux-Vekemans B, Journet A, André B, and Gasnier B. (2012). Heptahelical protein 
PQLC2 is a lysosomal cationic amino acid exporter underlying the action of cysteamine in cystinosis 
therapy. Proc. Natl. Acad. Sci. USA. 109, E3434-3443. 
 
[8] Yee DC, Shlykov MA, Västermark A, Reddy VS, Arora S, Sun EI, and Saier MH Jr. (2013). The 
transporter-opsin-G protein-coupled receptor (TOG) superfamily. FEBS J. 280, 5780-5800. 
 
[9] Ohsumi Y, and Anraku Y. (1981). Active transport of basic amino acids driven by a proton motive force 
in vacuolar membrane vesicles of Saccharomyces cerevisiae. J. Biol. Chem. 256, 2079-2082. 
 
[10] Sato T, Ohsumi Y, and Anraku Y. (1984). An arginine/histidine exchange transport system in 
vacuolar-membrane vesicles of Saccharomyces cerevisiae. J. Biol. Chem. 259, 11505-11508. 
 
[11] Li M, Rong Y, Chuang YS, Peng D, and Emr SD. (2015). Ubiquitin-dependent lysosomal membrane 
protein sorting and degradation. Mol. Cell. 57, 467-478. 
 


出芽酵母Saccharomyces cerevisiae液胞塩基性アミノ酸輸送系に関する研究

栗田, せりか

KURITA, Serika

（様式５）(Style5) 
Fig. 2. Structure of trans-resveratrol (1) 
学位論文全文に代わる要約 
Extended Summary in Lieu of Dissertation 
 
 
氏名： 
Name 
 栗田 せりか 
 
学位論文題目： 
Title of Dissertation 
Study on the Functionality of Polygonum Cuspidatum 
（イタドリの機能性に関する研究） 
 
 
学位論文要約： 
Dissertation Summary 
 
 Polygonum cuspidatum, commonly known as Japanese knotweed, is a rhizomatous perennial geophyte 
of family Polygonaceae and is originated in East Asia including Japan, Korea and China (Fig. 1) [1]. P. 
cuspidatum has distributed worldwide, and it is considered as one of the most invasive species [3]. Even though P. 
cuspidatum is viable and invasive, this plant is widely utilized by people in the East Asian countries. Its rhizome, 
Hu Zhang, is a traditional Chinese medicine and used to treat various illnesses which include infectious diseases 
as well as lifestyle-related diseases [7, 8]. Numbers of bioactive compounds 
have been identified to explain such effects. One of the notable compounds 
found in this plant is resveratrol (trans-3,5,4'-trihydroxystilbene) (1) (Fig. 2). 
Resveratrol with its various health promoting effects have become popular 
among people in modern society, and P. cuspidatum rhizomes which 
abundantly contain resveratrol are utilized as a source for resveratrol 
supplements [19]. On the other hand, the above ground parts such as stems and 
leave are recognized as food ingredients in Japan, especially in Kochi where 
the stems are traditionally pickled and cooked as a familiar food. Although its 
rhizomes have been well-researched for their effectiveness, the above ground, 
edible parts have rarely been the subject of study, so that there are few studies 
to evaluate the functionality of those parts of P. cuspidatum. In this study, the 
functionality of these edible parts of P. cuspidatum were investigated and the 
bioactive compounds related to such effects were elucidated. 
 
 
 
Quantification of resveratrol contents 
 
 To evaluate the functionality of the edible parts, the resveratrol content in each part of P. cuspidatum was 
first quantified. The fresh samples were collected from three different locations in Kochi prefecture (Shimanto, 
Kagami, and Muroto) in each season, and regional and seasonal 
differences in contents were observed. P. cuspidatum was separated into 
7 different parts, and the extracts were prepared with 80% MeOH 
extraction, liquid-liquid partition, and octadecylsilane (ODS) Sep-pak® 
cartridge. High performance liquid chromatography (HPLC) analysis 
was performed to see the amount of free resveratrol and total resveratrol 
which was obtained by hydrolysis using β-glucosidase. As a result of 
the study, the edible part was found to contain small amount of 
Fig. 1. Polygonum cuspidatum 
（様式５）(Style5) 
resveratrol (Table 1). In the edible young stem collected in spring, a trace amount of resveratrol was detected 
(~0.40 µg/g f.w.eq.). The leaf part contained resveratrol up to 7.35 µg/g f.w.eq.. The underground parts, tuber and 
rhizome, contained significant amount of resveratrol (~5260 µg/g f.w.eq.). The content of resveratrol changed by 
seasons and the tendency was different depending on the parts and the grown place. The above-ground parts 
showed increased amount in fall whereas the subterranean parts tended to increase in spring. Among three 
different regions, Muroto had a different tendency compared with other regions, probably due to difference in its 
weather and climate. This result suggested that the environmental factors greatly affect to the content of 
resveratrol as they do for other polyphenols. The antioxidant capacity of resveratrol and its contribution were 
measured using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity assay and superoxide anion 
scavenging activity assay. As an antioxidant, resveratrol contributed very little to the above-ground parts (< 
0.01%). Even in the tuber and rhizome which contained significant amount of resveratrol, its contribution was 
found to be small (1.9% and 4.0%, respectively), indicating the presence of other antioxidants which are greatly 
contributing to each part of P. cuspidatum.  
 
 
Table 1. Content of free trans-resveratrol and trans-resveratrol glycoside in each part of P. cuspidatum 
  
Content [µg/g f.w.eq.] 
Spring  Summer  Fall  Winter 
  free glycoside  free glycoside  free glycoside  free glycoside 
Shimanto             
Stem Whole  -  0.36 0.19  0.48 tr   - 
Lower tr 0.11   -   -   - 
Middle tr 0.17   -   -   - 
Upper tr 0.26   -   -   - 
Skin 0.18 0.10   -   -   - 
Leaf 
 
2.37 4.98  0.43 1.46  1.04 0.05   - 
Subterranean Tuber 744 2300  147 800  211 3036  360 2162 
Rhizome 1230 2690  61.5 1094  92.5 2036  97.8 780 
Kagami             
Stem Whole  -  tr 0.44  0.12 tr   - 
Lower tr 0.25   -   -   - 
Middle tr tr   -   -   - 
Upper tr 0.40   -   -   - 
Skin 0.21 0.24   -   -   - 
Leaf 
 
0.23 3.33  1.18 0.87  1.40 tr   - 
Subterranean Tuber 21.0 284  714 55.9  181 1420  53.7 937 
Rhizome 2120 3140  1010 84.1  302 1940  91.7 1450 
Muroto             
Stem Whole  -  tr 0.23  0.15 0.48   - 
Lower tr 0.34   -   -   - 
Middle tr 0.19   -   -   - 
Upper tr 0.39   -   -   - 
Skin tr 0.33   -   -   - 
Leaf 
 
1.95 0.42  2.24 3.65  2.79 2.44   - 
Subterranean Tuber  -  75.4 1230  39.2 1600  67.3 987 
Rhizome  -  177 2200  109 590  248 2270 
 
Note: “-” indicates not determined. “Whole” indicates the combination of lower, middle, upper and skin parts. “tr” indicates 
the content less than 0.1 µg/g f.w.eq. 
 
（様式５）(Style5) 
 
 
Antioxidant Activity of P. cuspidatum 
 
 Antioxidant capacity of the rhizome, 
stem and leaf of the plant was measured using 
DPPH radical scavenging activity assay, and the 
leaf showed the highest activity among three 
parts. Even well-known antioxidant, resveratrol, 
contributed less than 0.01% in the leaf; 
therefore, the edible leaf part was further 
fractionated in order to find out its predominant 
antioxidant. The predominant antioxidant was 
present in the ODS 20% MeOH fraction of the 
aqueous layer (Fig. 3, 4). The antioxidant was 
determined to be 3-caffeoylquinic acid 
(neochlorogenic acid) (2) which consisted of a 
quinic acid and caffeic acid ester as a result of 
nuclear magnetic resonance (NMR) and liquid 
chromatography-mass spectroscopy (LC-MS) 
analysis. In 1 g of the flesh P. cuspidatum leaves, 2.31 mg of neochlorogenic acid was contained which accounted 
for 12.8% of the total phenolic content of the leaves measured by Folin-Ciocalteu method. The antioxidant 
contribution was found to be 16.5% of trolox-equivalent (TE) antioxidant capacity (TEAC) and 36.5% of 
superoxide anion scavenging activity (SOSA) as examined by DPPH radical scavenging assay and superoxide anion 
scavenging assay, respectively. This result indicated that neochlorogenic acid works for a large part of SOSA among all 
other existing antioxidants. Neochlorogenic acid is a well functional compound, and P. cuspidatum leaves may be a 
good source for this compound. 
 
Fig. 3. Fractionation of leaf extract and TEAC of each fraction 
Fig. 4. HPLC profile for 20% MeOH fraction of aqueous layer of leaf extract 
（様式５）(Style5) 
 
 
Tyrosinase Inhibitory Activity of P. cuspidatum 
 
 Tyrosinase inhibitory activity was tested using 
mushroom tyrosinase. The leaf crude extract of P. 
cuspidatum showed inhibitory activity at 0.1 g/g f.w.eq. 
(63.0%). According to Lineweaver-Burk plot, the 
inhibitors from the leaf extract were speculated to be 
non-competitive inhibitors (Fig. 5). The leaf crude extract 
was further fractionated to elucidate the tyrosinase 
inhibitors. Upon the liquid-liquid separation, the inhibitory 
activity was observed in both BtOH and aqueous layers; 
however, inhibitors in BtOH layer was elucidated due to 
the difficulty in fractionation of the aqueous layer. The 
inhibitory activity was seen in the ODS 20% MeOH 
fraction of the BtOH layer and found to be smaller than 10 
kDa. In the structural analysis using NMR and LC-MS, 
the inhibitor found in this fraction was identified as 
luteolin 6-C-glucoside (isoorientin) (4) (Fig. 6), which has 
previously been reported to have melanogenesis. The 
content of isoorientin was 1.99 mg per 1 g of P. 
cuspidatum leaves, which is relatively high amount when compared with other plant leaves. Interestingly, the 
promoters of tyrosinase activity were also found in this fraction. These co-existing promoters also had flavone 
backbone in their structures and were identified as luteolin 8-C-β-glucopyranside (orientin) (3) and apigenin 
8-C-β-glucopranside (vitexin) (5) (Fig. 6). These flavonoids were structurally similar to isoorientin. However, 
they had glycosidic linkage at C-6 while isoorientin had glycosidic linkage at C-8, indicating that glycoside 
position may be the key to promote or inhibit the activity. The presence of isoorientin as an inhibitor of tyrosinase 
activity along with orientin and vitexin as promoters adds a new finding to P. cuspidatum research.  
 
 
 
 
 
 
 Fig. 6. Structures of orientin (3) (left), isoorientin (4) (middle), and vitexin (5) (right) 
Fig. 5. Lineweaver-Burk plot of tyrosinase activity with 
different concentration of aqueous layer of leaf extract 
（様式５）(Style5) 
 
Hyaluronidase Inhibitory Activity and Anti-allergic Effect of P. cuspidatum 
 
 The leaves of P. cuspidatum also exhibited 
hyaluronidase inhibitory activity. Strong activity (98.5%) 
was observed in the leaf crude extract at 50 mg f.w.eq./mL. 
To elucidate the active compound, the leaf crude extract 
was separated using liquid-liquid partitioning, ODS column 
chromatography and ultrafiltration. The inhibitor was found 
to be a > 10 kDa compound from ODS 50% MeOH 
fraction of the precipitate of the aqueous layer, and 
identified as proanthocyanidin (6) (Fig. 7), a polymerized 
polyphenol consisting of epicatechins with B type linkage 
of 4β→8 bonds. The number and weight average molecular 
weights of this compound were 2.82 × 104 and 6.97 × 104, 
respectively, and its mean degree of polymerization was 
determined to be 97.2. This highly polymerized compound 
exhibited uncompetitive inhibitory pattern, interfering with 
the enzyme-substrate complex in hyaluronidase activity 
(Fig. 8). The content of proanthocyanidins was 9.03 mg/g 
f.w.eq., and proanthocyanidins accounted for 24.6% of the 
entire inhibitory activity of P. cuspidatum leaves. Because 
hyaluronidase inhibitors often have suppressive effect on 
degranulation of mast cells or basophilic cells, the leaf crude 
extract and the isolated proanthocyanidins were tested for 
antigen-induced degranulation assay on RBL-2H3 cells. As 
a result, both the leaf crude extract and the isolated 
proanthocyanidins significantly reduced granulation of 
RBL-2H3 cells in a dose-dependent manner, indicating that 
they have alleviative effect on type I allergic symptoms (Fig. 
9). This is the first study to report about the hyaluronidase 
inhibitory and anti-allergic effect of P. cuspidatum leaves 
and about proanthocyanidins as effective compounds. 
 
Fig. 7. Structure of proanthocyandin (6) 
Fig. 8. Lineweaver-Burk plot for hyaluronidase activity 
with various concentrations of proanthocyanidins  
Fig. 9. Degranulation inhibitory effect of proanthocyanidins on sensitized RBL-2H3 cells (*p <0.05) 
* 
* 
（様式５）(Style5) 
 
 In conclusion, the functionality of the edible parts of P. cuspidatum was explored in this study, and 
especially the leaf part was found to possess various effects which can promote human health. Its antioxidant, 
tyrosinase inhibitory, hyaluronidase inhibitory activities as well as anti-allergic effect were evaluated, and the 
relating compounds were newly identified. Even though the leaves are not recognized as usable materials as the 
rhizomes, they can be as functional and valuable as the rhizomes. As a food ingredient or medicinal use, P. 
cuspidatum leaves potentially provides beneficial effects to our health in various ways. 
 
 
References: 
 
[1] Barney, J. N.; Tharayil, N.; DiTommaso, A.; Bhowmik, P. C., The biology of invasive alien plants in Canada. 5. 
Polygonum cuspidatum Sieb. & Zucc. [= Fallopia japonica (Houtt.) Ronse Decr.]. Canadian Journal of Plant Science 
2006, 86, 887-905. 
[2] Funkenberg, T.; Roderus, D.; Buhk, C., Effects of climatic factors on Fallopia japonica s.l. seedling establishment: 
Evidence from laboratory experiments. Plant Species Biology 2012, 27, 218-225. 
[3] Peng, W.; Qin, R.; Li, X.; Zhou, H., Botany, phytochemistry, pharmacology, and potential application of Polygonum 
cuspidatum Sieb.et Zucc.: A review. Journal of ethnopharmacology 2013, 148, 729-745. 
[4] Zhang, H.; Li, C.; Kwok, S. T.; Zhang, Q. W.; Chan, S. W., A review of the pharmacological effects of the dried root of 
Polygonum cuspidatum (Hu Zhang) and its constituents. Evidence-based Complementary and Alternative Medicine 
2013, 2013. 
[5] Szkudelska, K.; Szkudelski, T., Resveratrol, obesity and diabetes. European Journal of Pharmacology 2010, 635, 1-8. 
 
 
 
 


Study on the Functionality of Polygonum Cuspidatum

Ehime University (愛媛大学)博士(学術)thesi

Anastasia, Wheni Indrianingsih

（様式５）(Style5) 
学位論文全文に代わる要約 
Extended Summary in Lieu of Dissertation 
 
 
氏名： 
Name 
 
Anastasia Wheni Indrianingsih 
学位論文題目： 
Title of Dissertation 
 
Studies on Isolation and Characterization of Antioxidant and α-Glucosidase Inhibitory 
Compounds from Woody Plants and Endophytic Fungi 
(木本植物およびその内生菌からの抗酸化物質および α-グルコシダ―ゼ阻害
物質の単離と特性評価に関する研究) 
 
学位論文要約： 
Dissertation Summary 
 
Antioxidants are important since it protect the human body from free radicals and preventing 
diseases. Plants such as medicinal herbs, vegetables and fruits have been used as medicines since 
ancient time. Several studies demonstrated that medicinal herbs and plants are a rich source of 
antioxidant compounds such as phenolics, alkaloids and vitamins. Meanwhile, several synthetic drugs 
have been used for diabetes treatment since diabetes mellitus is the major health problem worldwide. 
However, several side effects such as drug resistane and weight gain had been observed. Some 
traditional medicines was demonstrated potential for diabetes treatment with less side effects, thus an 
effort to search natural antidiabetic medicines is important. The objective of this research was to search 
for antioxidant and α-glucosidase inhibitors from several woody plants.  The screening of antioxidant 
and α-glucosidase inhibitors was conducted and selected plants having the highest antioxidant and 
α-glucosidase inhibitory activity was then studied further for the isolation and characterization of the 
antioxidant and α-glucosidase inhibitor compounds.  Endophytic fungi from selected plants that 
having the highest antioxidant and α-glucosidase inhibitory activity were also studied.  
The antioxidant activity of the methanolic extracts of the 14 woody plants was determined 
using several assays: DPPH radical scavenging activity, H2O2 radical scavenging activity, β-carotene 
（様式５）(Style5) 
bleaching assay, and reducing power. The scavenging capacity of extracts ranged from 25.0 to >500 
µg/mL. Quercetin as a standard exhibited an IC50 of 8.4 µg/mL. Among the plant extracts, the highest 
DPPH scavenging capacity was shown by the extracts of Q. phillyraeoides A. Gray and Q. gilva 
Blume with an IC50 of 25.0 and 38.5 µg/mL, respectively. The scavenging capacity of extracts (IC50) 
on hydrogen peroxide ranged from 104.2 to >300 µg/mL. Q. phillyraeoides A. Gray exhibited the 
highest scavenging activity with 104.2 µg/mL, while gallic acid had a scavenging activity of 52.0 
µg/mL. The reducing power of the plant extracts ranged from 18.4 to 99.9 µg/mL. Overall, the results 
indicate that the subtropical plant Q. phillyraeoides A. Gray had the strongest reducing power of the 
extracts investigated, with 99.9 mg/g (gallic acid equivalent). Q. phillyraeoides A. Gray, Q. gilva 
Blume and M. japonicus had the highest ability in protecting β-carotene bleaching compared with 
other extracts with an activity of 52.1%, 52.4%, and 51.2%, respectively. These results were higher 
than the ascorbic acid as standard that has antioxidant ability of 25.2%. M. japonicus and Q. 
phillyraeoides A. Gray had the highest α-glucosidase inhibitory activity with an IC50 of 8.4 and 9.8 
µg/mL, respectively. The difference in these values was not statistically significant with quercetin as 
standard, which had an α-glucosidase inhibitory activity (IC50) of 4.2 µg/mL. The results of the present 
work showed that Q. gilva Blume, Q. phillyraeoides, and M. japonicus are potentially rich sources of 
natural antioxidants and anti-diabetes medicine. 
Quercus gilva Blume of the family Fagaceae is a tall evergreen tree distributed in the lowland 
mountain regions of Jeju Island in Korea. Moreover, Q. gilva Blume as an oak species in warm 
temperate regions also grows in Japan, in which it is mainly distributed in the southern part of the 
country. In the present study, the antioxidant and α-glucosidase inhibitor activities of isolated 
compounds from the leaves of Q. gilva Blume were tested. An in vitro assay of α-glucosidase 
（様式５）(Style5) 
inhibitory activity was conducted using α-glucosidase enzyme from S. cerevisiae yeast while an in 
vitro antioxidant activity assay was conducted using several methods: DPPH free radical scavenging, 
β-carotene bleaching assay, H2O2 radical scavenging assay, and reducing power assay. 
The methanol soluble fraction in the methanolic extract of Q. gilva Blume was fractionated using 
silica gel chromatography and followed by recrystallization to give compounds 1, 2, and 3 (Fig. 3.2). 
The 1H NMR and 13C NMR spectral data of all isolated compounds were compared with reported 
data, and their structures were identified as catechin (1), epicatechin (2), and tiliroside (3). 
The scavenging capacity of isolated compounds in DPPH free radical scavenging activity 
ranged between 22.55 and 160.24 µM. Of the isolated compounds, the highest DPPH scavenging 
capacity was shown by compound 2, followed by compounds 1 and 3 with IC50 of 22.55, 40.86, and 
160.24 µM, respectively. Quercetin and gallic acid were used as positive controls in this experiment 
and exhibited an IC50 of 28.08 and 20.01 µM, respectively. Compound 2 exhibited the highest 
scavenging activity on hydrogen peroxide assay of 116.71 µM, while gallic acid as a standard had 
scavenging activity of 308.24 µM. Compounds 1 and 3 had scavenging activities of 122.41 and 119.95 
µM, respectively. The reducing power of isolated compounds ranged between 41.60 and 61.47 µg/mL 
in gallic acid equivalents and between 78.77 and 98.96 µg/mL in ascorbic acid equivalents. Overall, the 
results indicate that compound 2 had the strongest reducing power among the isolated compounds 
investigated, with 61.47 mg/g (gallic acid equivalent) and 98.96 mg/g (ascorbic acid equivalent). The 
antioxidant activities of compounds 1 to 3 from Q. gilva at 40 µg/mL in the β-carotene-linoleate model 
system resulted in compound 1 having the highest ability in protecting β-carotene bleaching followed 
by compound 2 and compound 3, which still retained antioxidant activities of 14.93%, 10.44%, and 
1.49%, respectively, after 60 min of the assay. These results were higher than that for ascorbic acid as 
（様式５）(Style5) 
the standard, which had an antioxidant ability of 1.47%. Compound 3 had the highest α-glucosidase 
inhibitory activity (IC50) of 28.36 µM, followed by compounds 1 and 2 with 168.6 and 920.6 µM, 
respectively. These results for α-glucosidase inhibitory activity (IC50) were consistent with previous 
findings on catechin and epicatechin in green tea. Compound 1 and compound 2 exhibited an 
uncompetitive type of inhibition (Fig. 3.5), as shown by the straight parallel lines in the plot of 1/V 
versus 1/[S]. The Ki (inhibition constant) values of compounds 1 and 2 were determined to be 129.03 
and 215.05 μM, respectively (Table 3.2). Compound 3 showed non-competitive inhibition, which 
indicated that it bound to a site other than the active site of the α-glucosidase enzyme (Fig. 3.6a) with a 
Ki value of 91.64 μM. In conclusion, three compounds were isolated from the leaves of Q. gilva Blume. 
Of the isolated compounds, catechin (1) and epicatechin (2) showed potent antioxidant activities, while 
tiliroside (3) and catechin (1) showed potent α-glucosidase inhibitory activities. 
Quercus phillyraeoides A. Gray (Q. phillyraeoides) of the family Fagaceae is an evergreen 
tree that is distributed in the limestone mountains and acid bed rocks of East Asia (Korea, China and 
Japan). The leaves of Quercus species have been used in Korean folk medicine for dysentery, diarrhea, 
hemorrhage, dermatitis, and the exclusion of extravasated blood. Previous phytochemical studies on Q. 
phillyraeoides led to the identification of several tannins from the leaves of Q. phillyraeoides. However, 
the bioactivities of Q. phillyraeoides have not yet been examined. The present study was conducted in 
order to evaluate the antidiabetic and antioxidant potentials of constituents isolated from Q. 
phillyraeoides. Antidiabetic properties were assessed in terms of the ability to inhibit an intestinal 
carbohydrate-digesting enzyme, namely α-glucosidase. 
The methanol-soluble fraction in the methanolic extract of Q. phillyraeoides was fractionated 
using silica gel column chromatography and followed by recrystallization to give compound 4, while 
（様式５）(Style5) 
Sephadex LH-20 column chromatography led to the isolation of condensed tannin fractions 5 to 9. 
Compound 4 was identified as β-sitosterol-D-glucoside. The acid hydrolysis of 4 resulted in 
β-sitosterol and D-glucose. Fractions 5 to 9: light brown powders. The acid-butanol assay of fractions 5 
to 9 resulted in red colored anthocyanidin, which indicated that the fractions were condensed tannins 
(Schofield et al., 2001). The retention time of fractions 5 to 9 on GPC were 6.81, 12.78, 15.36, 10.55, 
and 15.87 min, respectively, therefore the molecular weights of fractions 5 to 9 were 5700, 4200, 3500, 
4700 and 3400 g/mol, respectively, which were estimated using polystyrene as the standard. The 
condensed tannin fractions (5 to 9) had high α-glucosidase IC50 values in the range of 2.60 to 3.14 
µg/mL, and no significant difference was observed from that of quercetin as the standard (P < 0.05) 
with an IC50 value of 4.81 µg/mL. Compound 4 showed moderate inhibitory activity against 
α-glucosidase with an IC50 value of 118.8 µg/mL, which was significantly lower than those of 
quercetin and the other isolated fractions. The α-glucosidase inhibitory activities of fractions 5 to 9 
were higher than that of compound 4, and this may have been because they consisted of more hydroxyl 
groups. The inhibitory mechanisms of the isolated constituents were analyzed further using 
Lineweaver-Burk plots. The results obtained showed that all of the isolated constituents and quercetin 
as the standard exhibited a mixed type of inhibition. 
The antioxidant activities of the six isolated constituents were determined using several assays: 
DPPH free radical scavenging activity, hydrogen peroxide radical scavenging activity, reducing power, 
and β-carotene-linoleate model assays. The scavenging capacities of the isolated constituents in the 
DPPH free radical scavenging activity assay ranged between 9.34 and > 500 μg/mL. Of the isolated 
constituents, the highest DPPH scavenging capacity was exhibited by fraction 8, followed by fractions 
6, 5, 9, and 7 with IC50 values of 9.34, 10.53, 10.84, 12.98, and 13.12 μg/mL, respectively. Quercetin 
（様式５）(Style5) 
and gallic acid were used as positive controls in this experiment and had IC50 values of 8.45 and 3.41 
μg/mL, respectively. The weakest activity was observed for compound 4 with an IC50 value of > 500 
μg/mL in the DPPH free radical scavenging activity assay, which was significantly lower than those of 
the standard and other isolated fractions. The scavenging abilities of the isolated constituents on 
hydrogen peroxide were compared with gallic acid as the standard. Fraction 9 exhibited the highest 
scavenging activity (IC50) of 84.05 μg/mL, followed by fractions 7, 6, 8, 5, and compound 4 with IC50 
values of 84.51, 86.67, 86.94, 89.28, and > 500 μg/mL, while gallic acid as the standard had 
scavenging activity of 52.36 μg/mL. Based on a statistical analysis, no significant differences were 
observed in the IC50 values of fractions 5 to 9. The reducing powers of the isolated constituents ranged 
between 19.34 and 54.35 μg/mL in gallic acid equivalents and between 56.16 and 91.72 μg/mL in 
ascorbic acid equivalents (Fig. 4.5a). The results indicated that fractions 5 to 9 actively reduced Fe3+ to 
Fe2+ with values of 52.59, 54.35, 51.72, 53.47, and 54.05 mg/g (gallic acid equivalent) and 89.94, 
91.72, 89.05, 90.83, and 91.42 mg/g (ascorbic acid equivalent), respectively, while compound 4 
showed the lowest activity at 19.34 mg/g (gallic acid equivalent) and 56.16 mg/g (ascorbic acid 
equivalent). These potential reducing powers may have been due to the presence of the dihydroxy type 
of benzene derivatives, catechin and epicatechin, which are integral parts of condensed tannins. The 
antioxidant activities of compound 4 and fractions 5-9 from Q. phillyraeoides at 40 μg/mL in the 
β-carotene linoleate model system resulted in fraction 9 having the strongest ability to protect against 
β-carotene bleaching, followed by fractions 6, 8, 7, 5, and compound 4, which still retained antioxidant 
activities of 20.01%, 25.78%, 22.10%, 18.42%, 14.73%, and 1.05%, respectively, after 120 min of the 
assay. The results obtained for fractions 5 to 9 were higher than that of gallic acid as the standard, 
which had an antioxidant ability of 6.32%. In conclusion, β-Sitosterol-D-glucoside (4) and five 
（様式５）(Style5) 
condensed tannin fractions (5, 6, 7, 8, and 9) were isolated from the leaves of Q. phillyraeoides. Of the 
isolated constituents, condensed tannin fractions (5 to 9) showed potent α-glucosidase inhibitory 
activities and antioxidant activities, while β-sitosterol-D-glucoside (4) showed moderate inhibitory 
activity against α-glucosidase. 
Several research indicate endophytic fungi have bioactive compounds that could potentially be 
applied in various applications such as antioxidant, antifungal, antiviral, antibacterial and cyotoxic. In 
our previous study, we found that several bioactive compounds were isolated from Quercus gilva 
Blume and Quercus phillyraeoides A. Gray therefore in this study we conducted screening of 27 
endophytic fungi from Quercus gilva Blume and Quercus phillyraeoides A. Gray. Endophytic fungi 
QGS 01 from Q. gilva was found to have a strong α-glucosidase inhibitory activity. This QGS 01 
fungus was identified as a Xylariaceae sp. In this study, the isolation of α-glucosidase inhibitors from 
the ethyl acetate extract of this fungus was reported. 
The ethyl acetate extract of QGS 01 fungus mycelium was fractionated using silica gel 
chromatography to give constituents 10 and 11. Their spectral data were compared with reported data 
and their structure were identified as isocoumarin derivative (10) and fatty acid fraction (11). 
Compound 10 was identified as 8-hydroxy-6,7-dimethoxy-3-methylisocoumarine. Constituents 11 
(fatty acid fraction) was isolated as major active constituents of Xylariaceae sp and was the most active 
constituents on α-glucosidase inhibitory activity. Analysis using GC-MS and comparison with 
authentic standards showed that constituents 11 had two saturated fatty acids, palmitic acid (C16:0) and 
stearic acid (C18:0) and three unsaturated fatty acids, namely oleic acid (C18:1), linoleic acid (C18:2) 
and linolenic acid (C18:3). The α-glucosidase activity of compound 10 and constituents 11 (IC50) 41.75 
and 5.95 μg/mL, respectively.  The fatty acids constituent (11) consists of 33.7% of linoleic acid, 
（様式５）(Style5) 
29.8% of oleic acid, 24.1% of palmitic acid, 7.5% of stearic acid and 3.6% of linolenic acid. These 
results indicated that fatty acids with a double bond showed stronger inhibition than fatty acids without 
double bond. It is suggested that the existence of a double bond was important factor of inhibition 
α-glucosidase acitivity. In conclusion, endophytic fungi Xylariaceae sp QGS 01 was isolated from the 
stem of Quercus gilva Blume and showed the highest α-glucosidase inhibitory activity. Two active 
constituents, 8-hydroxy-6,7-dimethoxy-3-methylisocoumarine (10) and fatty acid fraction (11), were 
isolated from endophytic fungi Xylariaceae sp QGS 01. The results of the present work showed that Q. 
gilva Blume, Q. phillyraeoides A. Gray and endophytic fungi Xylariaceae sp QGS 01 from Q. gilva 
Blume are potentially a rich source as antidiabetic medicine. 
 
(注) 要約の文量は，学位論文の文量の約１０分の１として下さい。図表や写真を含めても構いません。
(Note)  The Summary should be about 10％ of the entire dissertation and may include illustrations 


Studies on Isolation and Characterization of Antioxidant and α-Glucosidase Inhibitory Compounds from Woody Plants and Endophytic Fungi

高橋, 尚志

TAKAHASHI, Shoji

アルミノケイ酸塩ガラスの特性と構造に及ぼすAL_2O_3の影響

寺谷, 亮司

愛媛大学地域創成研究センター活動報告－平成 26 年度（2014 年 4 月～2015 年 3 月） 
 
 2014 年度の地域創成研究センター教員は，センター長・寺谷亮司教授，副センター長・
和田寿博教授，佐藤亮子准教授，川岡勉教授（教育学部），松野尾裕教授（教育学部），野
崎賢也准教授（法文学部）の６名であり，前３者が当センター専任教員である。2014 年度
は，下記２部門において，多くの活動を実施した。  
 １．学術文化創造部門  
 各センター教員は，地域に根ざした研究や教育効果を意識した学生との協同調査を実施
している。前者には，「松山空襲など戦争体験記録プロジェクト」などがある。後者の成果
の一部は，日振島調査報告など，本年度に刊行した地域創成研究年報第 10 号にも掲載され
ている。また，今年度誕生した「松山アーバンデザインセンター主催の「U30 都市計画－
都市設計競技 風景づくり夏の学校 2014－」（道後温泉都市計画コンテスト）に学生とチー
ムを組んで応募し，そのポスター成果は道後温泉商店街や坂の上の雲ミュージアムで展示
された。  
 共同研究の推進・募集と支援に関しては，「えひめ３１１」，「愛媛グローバルネットワー
ク」などの NPO との交流活動を実施し，また「四国遍路 と世界の巡礼研究会」などの講
演会を支援・後援した。  
 地域の文化・芸術事業の推進としては，2 月 18 日に，「発酵茶の可能性を探る－世界に誇
る国内産地が愛媛に集結－」シンポジウムを開催し，全国から 100 名を超える参加者を得
た。  
 ２．地域政策部門  
 同部門におけるシンクタンク機能の推進に関しては，松山市との「道後温泉活性化ワー
クショップ開催・運営業務」，「道後温泉活性化懇談会」，「松山市清水地区まちづくり協議
会」，「松山市中心市街地活性化事業」，「松山市中小企業振興基本条例検討委員会」，「宇和
島市津島町御槇地区集落コミュニティ支援事業」，「上島町魚島地域元気アッププロジェク
ト」，「西条市市民憲章等制定事業」を継続・実施した。特に，「宇和島市津島町御槇地区集
落コミュニティ支援事業」では，「元気な集落づくり支援事業」を土台に立ち上がった「み 
まきガーデン」の運営を軌道にのせるための支援活動として，7・10・11・12・2 月にワー
クショップを実施し，地域住民と運営体制や事業内容についての検討を行った。  
 地域貢献事業では，「mit まちなか大学」を開催し，市民に向けた「知」の発信に努めた。
今年度は，第１回「松山中心部のオフィスビルの動向と課題」（12 月 18 日），第２回「愛媛
らしいお酒を考える－「しずく媛」酒を中心に－」（1 月 19 日），第３回「元気な島づくり
を考える」（1 月 27 日），第４回「愛媛 大学法文学部地域コース・観光まちづくりコースに
よるまちづくり報告会」（3 月 25 日）を内容とする講演会を実施した。  
 松山市との共同事業「地域リーダー養成セミナー」では，今年度は５～７月にカリキュ
ラムを集中的に実施する日程とし，7 月 16 日には清水公民館にて，地元住民の前で「ミニ
まち改善プロジェクト」成果を報告した。今年度の同事業の修了生は 37 名であり，地域人
材の養成に貢献した。 
                                                             （地域創成研究センター長 寺谷 亮司） 
110
地域創成研究年報第１０号（2015年）


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