Hachisuka Mochiaki's Overseas Achievements

蜂須賀茂韶の海外での功績

Hachisuka Mochiaki (1846-1918) was the last lord of the Awa (Tokushima) domain, and he became a successful businessman and a statesman after the Meiji Restoration. He went to the UK to study when he was 25 years old, and stayed there for seven years, during which he graduated from college at the Balliol College, University of Oxford. Three years after returning to Japan, he was appointed to an envoy extraordinary and minister plenipotentiary of Japan in France (also serving as a minister in Spain, Portugal, Switzerland and Belgium) and he stayed in Paris three years. Despite ten years of foreign life, little has been known about his overseas activities. In this study, we investigated his overseas activities using digital archives of European libraries, and could find information from British Newspaper Archive, Welsh Newspaper Online, and Gallica. He began to attend public events in the UK after graduating from the Oxford University. He received in an audience by Queen Victoria and the Prince of Wales in 1877 and 1878, and attended to parties hosted by the Ministry of Foreign Affairs. He also participated in the launching ceremony of three Japanese warships Kongo, Fuso, and Hiei in 1877. He worked variously as a diplomat in Paris during 1884-1886. He signed three treaties, the Geneva Convention, the treaty on remittance by postal money order, and the International Meter Convention. Conpared with signing the Geneva Convention remaining two have not received much attention so far. In this study we found that he encouraged the Japanese government to join the International Meter Convention and negotiated with the Comité International des Poids et Mesures many times. He organized Japanese exhibition at a museum to introduce Japanese culture to Parisians, and he contributed to the academic exchange between France and Japan with the Geographical Society. And he and his wife Yoriko participated in many social events, and they held parties, concerts and theaters in Paris and Brussels. Several French newspapers mentioned elegant behavior of his wife Yoriko at those parties. And He focused on politics and business after returning to Japan, but also worked to promote exchange with foreign countries. In particular, he established the Welcome Society, which was the first organization in Japan for attracting and accommodating foreign tourists, with other of founders, and became the chairman of the organization. His abundant overseas experience and wide-ranging personal connections helped to establish this association.

蜂須賀茂韶(はちすかもちあき)は，藩政期の終わりに徳島藩の最後の藩主となり，明治維新後は外交官，実業家，政治家として活躍した人物であるが，彼のイギリス遊学時代や外交官使時代の活動については，これまで情報が乏しかった．本研究はヨーロッパの図書館のデジタルアーカイブを利用して彼に関する情報を調べ，多数の資料を発見した．茂韶はオックスフォード大学を卒業してからもしばらくイギリスに滞在し，ヴィクトリア女王や皇太子に拝謁したり，軍艦「金剛」「扶桑」「比叡」の進水式に出席したりと公的な行事に現れるようになった．パリでの公使在任中の活動については多くの資料が見つかり，多彩な活動内容が明らかになった．条約関連では，ジュネーブ条約，日仏間郵便為替条約，メートル条約に署名した．この中ではジュネーブ条約署名がよく知られているが，メートル条約も日本の社会や産業に及ぼした影響は計り知れないものがある．国際度量衡委員会の活動報告書CIPM 1885(111)から，茂韶が国際度量衡委員会や日本政府と何度もやりとりを重ねて条約加盟にこぎつけたことが明らかになった． また，デジタルアーカイブには古新聞が多く含まれているため社交界の情報も多く，茂韶夫妻が多数の催しに出席したことが分かった．夫妻は主催することにも熱心で，ベルギーのブリュッセルで開いた舞踏会や，パリでの晩餐会やコンサートや観劇は評判になった．

Regional Science Research, the University of Tokushima


Wenceslau de Moraes's Proficiency of Japanese Language --- His Speaking Ability and Personality

ヴェンセスラウ・デ・モラエスの日本語会話能力 --- 会話能力の検証および会話内容からみえる人物像について

A Portuguese writer Wenceslau de Moraes spent his later life in Japan since 1897; he never left Japan until 1929 when he died. It is said that his speaking ability of Japanese language was not good although he lived in Japan more than thirty years. In this study, we investigated his speaking ability of Japanese based on the written memories of those who had direct discourse with him. The records of his speech were limited to the memories of his neighbors, a Buddhist nun who regularly came to his house, and a newspaper reporter from Osaka who interviewed him. Except for the words of affirmation or denial, the word that appeared most frequently in his speech was "kawaisou'' or ``kinodoku'' that means ``pity'' (``piedade'' in Portuguese). ``Piedade'' is the important key word of his masterpiece Ó-Yoné e Ko-Haru. In the preface to the book he quoted the sentence ``A litteratura do futuro será a Litteratura da piedade'' from Pierre Loti's work, and he wrote Ó-Yoné e Ko-Haru as a literature of pity. Therefore, his deep sympathy for vulnerable and oppressed existence was the propensity of his character shown in both his works and his real life. ``Kawaii / kawairashii'' (lovely) and ``shinsetsu'' (kind) are also the words he used frequently in his conversation with his neighbors. As for the grammar of Japanese language, Moraes did not seem to have mastered postpositional particles and conjugation of verbs. Two persons referred to his speaking ability of Japanese as follows: a newspaper reporter who interviewed Moraes wrote that ``he speaks in simple Japanese'' in his article, and a pastor who sometimes visited Moraes's house said that ``his Japanese was not good, but he spoke familiar Japanese slowly''. The reason why Moraes's Japanese did not improve is said that he did not want to associate with intellectual people here in Tokushima. On the other hand, he greeted familiarly and often exchanged gifts with his neighbors. He pointed out that Japanese people is a pleasant neighbor in his work Relance da Alma Japoneza, and he also tried to be a good neighbor to those he knew as well. There would have been no need for a formal and complicated conversation for him for that purpose.

ヴェンセスラウ・デ・モラエスは，1897年以来，日本に30年以上日本に住んでいたにもかかわらず，日本語の話す能力は良くなかったと言われている．本研究では，直接会話をした人々の記憶に基づいて，彼の日本語の話す能力を調査した．肯定や否定の言葉を除いて，彼の発話で最も頻繁に現れた言葉は「かわいそう」「気の毒」であった．ポルトガル語ではpiedadeが対応する言葉であり，彼の著作の中でも頻繁に現れ，彼を理解する上で重要なキーワードである．また 「カワイイ/カワイらしい」や「しんせつ」なども，会話によく使われた．日本語の文法に関しては，モラエスは助詞や動詞の活用形を習得していなかった．モラエスの日本語が上達しなかったのは，彼が徳島で知的な人々との付き合いを避けたためであろう．その一方で，彼はよく挨拶し，彼の近所の人たちと贈り物をしばしば交換していた．彼は日本人が心地よい隣人であることを自分の作品の中で述べており，彼もまた良い隣人であろうと努めた．彼の生活において知的で複雑な会話は必要なかったのである．


First report of `Candidatus Phytoplasma malaysianum' associated with Elaeocarpus yellows of Elaeocarpus zollingeri

``Elaeocarpus yellows'' (ELY) is a widely reported phytoplasma disease of Elaeocarpus zollingeri trees in Japan. The phytoplasma associated with ELY (ELY phytoplasma) had not been identified at the species level because its 16S rRNA sequence had yet to be reported. Here, we report the results of a sequence analysis based on 16S rRNA and secA gene sequences, which showed that the ELY phytoplasma is related to `Candidatus Phytoplasma malaysianum'. To our knowledge, this is the first report showing the occurrence of `Ca. P. malaysianum' outside Malaysia and the infection of E. zollingeri by the phytoplasma.


Present situation of Elaeocarpus zolloingeri tree planted in Mt. Shiroyama in 2006 --- survival and infection status of Elaeocarpus yellows

Elaeocarpus zollingeri (synonym: E. sylvestris) was one of dominant trees in Mt. Shiroyama in the Tokushima City until 1970s, however, most of adult trees have died by Elaeocarpus yellows. To prevent extinction 300 nursery trees were planted at the foot of the mountain by local volunteers in 2006. In this study we investigated their survival, growth, and infection status of the disease. We found 40 nursery trees have survived (survival rate was 13%). Survived trees were abundant in areas facing the south, and most trees disappeared in areas facing the north and the west. Growth of the trees was well also in areas facing the south. Among the survived 40 nursery trees, we found three trees infected with phytoplasma, the pathogen of Elaeocarpus yellows, based on PCR analysis. It is not clear when they were infected with phytoplasma, whether they have infected after the plantation or they already had phytoplasma in the seed. Anyway, the fact that only a few tree is infected must be gratifying in considering the conservation E. zollinger in Mt. Shiroyama.

2006年(平成18)6月25日にNPO「徳島城址を愛する会」の主催により城山山麓に植えられた300本のホルトノキの苗木の調査を行った．調査の結果，これらのうち40本の苗木の生存が確認された(13.3%)．エリア別にみると，南側山麓に比較的多く残っていた．また,植樹記録のない苗木も2本見つかった．これらがホルトノキ萎黄病の原因菌ファイトプラズマに感染しているかどうかをnested PCRにより確認した結果，3本がファイトプラズマに感染していることが明らかになった．

Regional Science Research, the University of Tokushima


Bridges and a Park Made by German Prisoners in The Bandō Prisoner-of-War (POW) Camp

板東俘虜収容所のドイツ兵が大麻比古神社境内に造った橋と公園

German prisoners in the Bandō prisoner-of-war (POW) camp made bridges and a park in the forest of the Ooasahiko-Jinja Shrine during 1917-1919. Details of the constructions were a 15 m-long wood bridge and five small wood bridges, four stone bridges, road with a total length of 1,130 m, stone embankments, slopes, two flights of stone steps with a length of 8 m and 3 m according to the record written by Adolf Deutschmann who planned and directed the work. Two stone bridges called Doitsu-bashi (German bridge) and Megane-bashi (glasses bridge) remain and the Doitsu-bashi has been designated as a cultural asset of Tokushima prefecture. However, other constructions have been disappeared or became unclear now. In this study, we tried to clarify their precise place and history using an old map of the Ooasahiko-Jinja Shrine, photos and pictures of German prisoners, and interview to an old resident who participated in improvement of the forest around 1970. Results indicated that the road and the stone bridge still remain in some part. In addition to Adolf Deutschmann we found that Max Bunge was an important member of the work. He has not been noticed in previous studies of the Bandō POW camp, although he was a famous person in the German community in Qingdao by some heroic behaviors, and that he became a mayor of his hometown Heiligenhafen after he was released from the camp. He wrote about beautiful nature of the shrine's forest and about his sympathy to faith of Japanese pilgrims who he saw during construction work.

鳴門市大麻町板東にある大麻比古神社の本殿裏の林内に「ドイツ橋(独逸橋)」，「めがね橋」と呼ばれる2つの石橋がある．これらは板東俘虜収容所に収容されていたドイツ兵が1917年から1919年にかけて橋造り・公園造りの一環として築いたものである．本研究はドイツ兵が造った公園がどのようなものだったのか具体的に明らかにすることを目的として行なった．ドイツ兵が残した記録や写真，地元の古老への聞きとり，大麻比古神社の古境内図等を基に，ドイツ兵が何をどこに造ったのか推測・推定した．そして，2つの石橋の他に，石段と道にドイツ兵が築いた名残を留めていることが分かった．また，調査の過程で，これらを設計し作業を指揮したドイッチュマン築城少尉の他にマックス・ブンゲ曹長が関わっていることが分かった．彼は，青島ドイツ人社会において英雄と称えられた人物であり，収容所から解放された後，帰国して故郷の町ハイリゲンハーフェンで町長を12年務めた．彼については今日まで板東俘虜収容所を題材にした一連の調査や作品群において注目されることがなかったが，このように傑出した人物であり，橋や公園の保存・管理にあたり彼の存在についても記憶に留めるべきであると考える．


Prevalence of Elaeocarpus Yellows in Mt. Shiroyama in Tokushima City

徳島市城山のホルトノキの衰弱・枯死の原因について — ホルトノキ萎黄病を引き起こすファイトプラズマの深刻な感染状況 —

Elaeocarpus sylvestris var. ellipticus was a dominant tree species in Mt. Shiroyama (castle mountain) in the Tokushima Central Park until 1970s, however, most of the E. sylvestris trees have died. In this study we investigated whether the death has been caused by Elaeocarpus yellows. Results of nested PCR revealed that DNA of phytoplasma, the pathogen of Elaeocarpus yellows, was found in all E. sylvestris trees in Mt. Shiroyama and about 80% E. sylvestris trees in the Tokushima Central Park. Results of PCR-RFLP showed that the all the trees had the same DNA type of phytoplasma, and it was identical to the one that had been found in Japan. These results indicated that the recent decrease of E. sylvestris in Mt. Shiroyama is due to Elaeocarpus yellows.

徳島市の「城山原生林」のホルトノキ群落は1980年代以降，急速に衰弱・枯死が進み，200本以上あった大径木が11本しか残っていない．その原因が全国的なホルトノキの枯死を引き起こしているホルトノキ萎黄病かどうかを検証した．その結果，分析した城山のホルトノキすべてにホルトノキ萎黄病の原因細菌であるphytoplasmaが検出され，ホルトノキ萎黄病の被害が深刻であることが明らかになった．


Triclocarban-induced change in intracellular Ca2+ level in rat thymocytes: Cytometric analysis with Fluo-3 under Zn2+-free conditions

Triclocarban (TCC) is an antimicrobial used in personal hygiene products. Recent health concerns arose after TCC was detected in the blood of human subjects who showered with soap containing TCC. In this study, the effect of TCC on intracellular Ca2+ concentration in rat thymocytes was examined using Fluo-3, an indicator of intracellular Ca2+. TCC at concentrations ranging from 0.1 µM to 3 µM increased intracellular Ca2+ concentration biphasically: first by releasing Ca2+ from intracellular Ca2+ stores and then inducing Ca2+ influx through store-operated Ca2+ channels. The threshold TCC concentration to increase intracellular Ca2+ concentration in this study was lower than the maximum TCC concentrations reported in human blood samples. Therefore, we anticipate that TCC at concentrations reported in human blood samples might disturb intracellular Ca2+ signaling in human lymphocytes.

Environmental Toxicology and Pharmacology


Induction of non protein thiols by chromium in cyanobacteria isolated from polluted areas

Two cyanobacterial strains Synechocystis ``AHZ-HB-MK'' and ``AHZ-HB-P2A'' were isolated from tannery effluent form Lahore, Pakistan and their tolerance against K2CrO4 was examined. Although growth was completely inhibited at 10 μg mL-1 in AHZ-HB-P2A strain, cell growth of AHZ-HB-MK strain was observed even at a concentration of 100 μg mL-1. To show reasons for different tolerance between the two strains, we compared changes in nonprotein thiols, glutathione (GSH and GSSG), cysteine and cystine, after K2CrO4 treatment in both strains. After K2CrO4 treatment GSH completely disappeared in both strains, suggesting that GSH was involved in detoxification. GSSG content in former strain remarkably increased almost three and six times after 7-days treatment with 10 and 100 μg mL-1 K2CrO4, respectively while it decreased in the later strain. Cysteine content in the former strain increased almost three and eleven times after 1 day and 7 days treatment with 100 μg m L-1 K2CrO4, respectively, such increase in cysteine content was not observed in later strain. These data revealed that former strain could induce glutathione and cysteine but later strain could not. The difference in the ability to induce the nonprotein thiols between two strains must be responsible for differential tolerance in K2CrO4 observed in the growth inhibition.

African Journal of Microbiology Research


Effect of triclocarban on membrane potential of rat thymocytes: Assessment with bis-(1,3-dibutylbarbituric acid)trimethine oxonol

The effect of triclocarban (TCC), an environmental pollutant from household items and health care products, on membrane potential of rat thymocytes was examined by a flow cytometry with a fluorescent probe sensitive to membrane potential, bis-(1,3-dibutylbarbituric acid)trimethine oxonol, because TCC changes intracellular ionic circumstance that may affect the membrane potential. TCC at 0.3 µM or more (up to 3 µM) depolarized the membranes. This TCC-induced phenomenon was against our prediction because TCC increases intracellular Ca2+ concentration that activates Ca2+-dependent K+ channels, resulting in a hyperpolarization. The depolarization was still observed under Ca2+-free condition, but not under Na+-free condition. Furthermore, TCC hyperpolarized the membranes under Ca2+- and Na+-free condition. To see if TCC inhibits Ca2+-dependent hyperpolarization, the effect of A23187, a calcium ionophore, on the membrane potential was examined in the cells treated with TCC. A23187 induced large depolarization in the cells treated with 0.3 3 µM TCC. The A23187-induced depolarization in the presence of TCC was greatly attenuated under Na+-free or Ca2+-free condition whereas A23187 elicited hyperpolarization in the cells treated with 0.3 3 µM TCC under Ca2+- and Na+-free condition. Results suggest that 0.3 3 µM TCC increases membrane permeability of Na+ and Ca2+, resulting in the depolarization. Large depolarization induced by TCC in the presence of external Ca2+ and Na+ may mask the hyperpolarization elicited via the increase in intracellular Ca2+ concentration by TCC. Thus, there is a possibility that TCC depolarizes membranes of lymphocytes, resulting in alteration of cellular functions of lymphocytes.


Synergistic increase in cell lethality by dieldrin and H2O2 in rat thymocytes: Effect of dieldrin on the cells exposed to oxidative stress

Dieldrin, one of persistent pesticides, is highly resistant to biotic and abiotic degradation. It is accumulated in organisms. Recent studies suggest that dieldrin exerts a potent cytotoxic action on cells exposed to oxidative stress. In this study, the effect of dieldrin on rat thymocytes exposed to hydrogen peroxide (H2O2)-induced oxidative stress was examined. Dieldrin at 5 µM and H2O2 at 300 µM slightly increased cell lethality from a control value of 5.4 ± 0.5% (mean ± standard deviation of four experiments) to 7.8 ± 1.3% and 9.0 ± 0.3%, respectively. Simultaneous application of dieldrin and H2O2 significantly increased cell lethality to 46.2 ± 1.8%. The synergistic increase in cell lethality was dependent on dieldrin concentration (0.3 5 µM) but not on H2O2 concentration (30 300 µM). Dieldrin accelerated H2O2-induced cell death, which was estimated with the help of annexin V-FITC and propidium iodide. Presence of either dieldrin or H2O2 decreased the cellular content of nonprotein thiol and increased intracellular Zn2+ concentration. The combination of dieldrin and H2O2 further pronounced these effects. TPEN, a chelator of intracellular Zn2+, significantly attenuated the synergistic increase in cell lethality induced by dieldrin and H2O2. It is, therefore, suggested that dieldrin augments the cytotoxicity of H2O2 in a Zn2+-dependent manner.


Nanomolar concentration of triclocarban increases the vulnerability of rat thymocytes to oxidative stress

It was recently reported that triclocarban was absorbed significantly from soap used during showering in human subjects and that its Cmax in their whole blood ranged from 23 nM to 530 nM. We revealed that a nanomolar concentration (300 nM) of triclocarban potentiated the cytotoxicity of 300 µM H2O2 in rat thymocytes by using cytometric techniques with appropriate fluorescent probes. Although 300 nM triclocarban did not itself increase the population of dead cells (cell lethality), it facilitated the process of cell death induced by H2O2, resulting in a further increase in the population of dead cells. Nanomolar concentrations (300 nM or higher) of triclocarban significantly decreased the cellular content of nonprotein thiol (glutathione), which has a protective role against oxidative stress. Triclocarban at 300 nM or higher increased the cell vulnerability to oxidative stress. The results may suggest that nanomolar concentration (300 nM or higher) of triclocarban affects some cellular functions although there is no evidence for adverse effects of triclocarban in humans at present.


Landscape Architecture of Tokushima Central Park --- Comparison with Hibiya Park and Seifersdorf Castle

徳島公園(徳島中央公園)の造園設計について --- 日比谷公園及びザイファースドルフ城との比較

Tokushima Park (originally named Tokushima Central Park) is the Japan's second western-style park that was opened in 1906. We investigated landscape architecture of Tokushima Park based on a blueprint made in 1905 to understand its purpose and function of the park, and compared with Hibiya Park that is the Japan's first western-style park. Tokushima Park consisted of five areas. The central area included Mt. Shiroyama (Castle Mountain), and primeval forest was protected without allowing to make a big building within the area. A commercial museum, an athletic field, and a botanical garden and a library were placed in the southern, western and eastern areas respectively, so that each area was designed to exhibit each function. Tokushima Park and Hibiya Park were designed by the same two persons Seiroku Honda and Takanori Hongo. The two parks were equipped with a wide road, an athletic field, a botanical garden and so on, and these facilities were adopted to the park made since them. Because Seiroku Honda adopted three design drawings of German parks from the book Gärtnerisches Planzeichnen into a blueprint of Hibiya Park, we investigated the book to ascertain whether any design drawing was also used in Tokushima Park. We found that Seifersdorf Castle, the castle of count Brühl that was built at Seifersdorf in Germany in 13th century, is similar to the southern area of Tokushima Park.

徳島中央公園は明治39年(1906)に旧徳島藩主の居城跡に「徳島公園」として開設された我が国で二番目の西洋風近代公園であり，市の中心部に位置する総合公園として多くの人々に利用され親しまれてきた．徳島公園について設計の詳細に踏込んだ研究はこれまで行われてこなかったが，それは設立前後の公園の設計図や図面が残っていないことによる．演者らは明治38年(1905)の新聞紙上に徳島公園の設計図やその解説が掲載されていることを見いだし，それに基づいて徳島公園設計の意図や理念を探った． 徳島公園の設計を担当したのは，我が国初の西洋風近代公園として作られた日比谷公園の設計を担当した本多静六と弟子の本郷高徳である．徳島公園と日比谷公園の設計図の調査から，日本における初期の西洋風近代公園の特質を浮き彫りにした． また，本多は日比谷公園の設計にあたり，造園家マックス・ベルトラムによるドイツの公園の設計図集『造園設計図案』(Gärtnerisches Planzeichnen)を取り入れている．そこで，徳島公園の設計に際してもこれを参考にしている可能性があると考え，調査した結果，XXXII番の``Die Parkanlagen des Herrn Graf Brühl zu Seifersdorf bei Radeberg''と題された図が徳島公園と類似していることが分かった．この図はドイツのザクセン州ラーデベルク近くのザイファースドルフ(Seifersdorf)にある13世紀に作られたBrühl 伯爵の城である． また，林学者であった本多は，城山について「天然林として甚貴重すべきもの」とその価値を認め，「林木の伐採は道路の開鑿及び眺望を得るための他は之を避け」と述べているように，本丸跡の招魂碑と東二の丸跡の喫茶店の他に建築物は設けなかった．さらに彼は「徳島公園は公園としての資質を備ふる点に於て蓋し日本の各公園中第一位」と述べ，その理由を三つ挙げているが，いずれも城山に関係したものである．

Regional Science Research, the University of Tokushima


Nature and landscape of Mt. Bizan based on ``Awa meisho zue''

『阿波名所図会』における眉山の自然と景観

In this paper the nature and the landscape of the Mt. Bizan are investigated based on ``Awa meisho zue'' which was a guidebook of Awa (Tokushima) published about 200 years ago. In the picture entitled ``Mt. Bizan'' a tower at the Jimyoin Temple and two buildings are drawn in the mountain. The mountain is covered with pine trees, and there is a waterfall near the tower. Besides pine trees, three types of trees, Japanese cedar and/or Japanese cypress, trees that presumably cherry, and unknown broadleaf trees are drawn in Mt. Otakiyama that is a part of Mt. Bizan based on the picture entitled ``Otakisan Jimyoin''. Other historical records in Edo era coincide with the composition of plant species in Mt. Bizan. Though the vegetation of Mt. Bizan was defended by laws throughout Edo era, it was deforested and the mountain became bald after the Meiji Restoration. Afterwards, it was protected by specifying it as the protection forest and by making it as a park. Now, pine trees and Japanese cedar and/or Japanese cypress have almost been lost, and pasania and live oak are well growing in the mountain. The tower at Jimyoin was burned down by the air raid in the World War II, and the waterfall has thinned. On the other hand, mountain trails and a ropeway were made, and a lot of buildings were built at the top of the mountain. Thus, the nature and the landscape of Mt. Bizan greatly changed in 200 years. However, citizens are still familiar with it as the symbol of Tokushima City.

『阿波名所図会』は，江戸時代後期に庶民の間で旅行ブームが興った時期に作られた阿波の名所を紹介したガイドブックである．同書巻頭の附言に「文化辛未之歳中冬」とあることから，文化8年(1811)に作成されたことが分かり，今年はちょうど200年目にあたる．本研究は『阿波名所図会』において眉山の景観や自然がどのように描かれているか分析するとともに，それ以降現在までの変化についても他の資料を基に調査した． 『阿波名所図会』において眉山が描かれた挿絵には，山中の樹木が4種類に描き分けられている．最も目立つのは，山中に生い茂っている松である．次に，松とは異なる針葉樹が多数描かれているが，これと同じ樹は同書にしばしば登場し，挿絵「舎心山太龍寺」では山中にこの樹が優占している．太龍寺の境内は現在，杉や檜の古木で覆われており，『阿波名所図会』においても杉または檜を描いていると考えられる．江戸時代に作成された別の史料『四国偏礼霊場記』や『四国遍札名所図会』との比較からも杉・檜であることが支持される．他の2種類の樹木は広葉樹であり，形態的特徴から樹種を特定することは難しいが，他の挿絵や別の史料との比較からそのうちの一つは桜であると考えられる． このような眉山の植生景観は明治期に一変する．大正5年(1916)版の『阿波名勝案内』には，明治維新後に眉山の樹木が伐採されて禿げ山となったことが記されている．その後，大正期から太平洋戦争前の間に作られた古絵葉書「徳島・新町橋」を見ると，再び眉山は木々で覆われている．樹種は判然としないが，広葉樹が主であり，松や杉・檜は目立たたない．また，同時期の別の古絵葉書「新町橋と眉山」では，山腹に白く桜並木が，筋状に延びているのが分かる． 平成16年の『徳島県環境基本計画(資料編)』には現在の眉山は，山地のアカマツ林はほとんど枯れてコナラ林に変わったと記されている．明治維新後の伐採で大きく数を減らした眉山の松は，マツ枯れによりさらにその数を減らしたのである．同資料は眉山の自然植生を，ヤブツバキ域にまとめられる照葉樹林「シイ - タブ林」に含まれる「ミミズバイ - スダジイ群集」に分類している．かつて眉山に広がっていた松林は，人間の影響下で自然植生の代償植生として成立していたものである． このように眉山の植生は，人為的影響やマツ枯れにより時代によって大きく変化してきた．現在は保安林，風致地区，鳥獣保護区に指定され，将来に残すべく保護対策がとられている．


Giant trees of Japan and the Korean Peninsula: comparison of species and folklore

日本と朝鮮半島の巨樹 – 樹種および巨樹にまつわる伝承の比較

Dai-Nippon Rouju Meiboku-shi is a book on trees in Japan published in 1913, when a trend to protect natural monuments was prevailing. It contains the address, trunk circumference at 1.5 m above ground, height, age, and legends and manners of 1500 famous trees. Chosen Kyoju Rouju Meiboku-shi, published in 1919, is a similar book of 3168 trees in the Korean Peninsula. In this study we tried to understand trees of Japan and the Korean Peninsula botanically and culturally based on these books. The major trees in Dai-Nippon Rouju Meiboku-shi are Pinus spp., Cryptomeria japonica, Cinnamomum camphora, Zelkova serrata, Prunus subg. cerasus, and Ginkgo biloba, and those in Chosen Kyoju Rouju Meiboku-shi are Zelkova serrata, Celtis sinensis and Aphananthe aspera, Ginkgo biloba, Pinus densiflora, Fraxinus mandshurica, and Styphnolobium japonicum. Major owners of the trees are shrines and temples in Japan, and regional communities in the Korean Peninsula. Legends of the trees vary depending on the species and seem to have partly originated from their morphological and physiological features. Moreover, comparison of the folklore about the trees brought out the difference in history, religion, and culture between Japan and the Korean Peninsula.

明治時代末の天然記念物保護の機運の高まり受け，大正2年(1913)に刊行された『大日本老樹名木誌』は，各地の著名な樹1500本について，所在地，地上五尺の幹周囲，樹高，樹齢，伝説が記されており，日本の巨樹研究において極めて重要な資料である．また，その6年後の大正8年(1919)には朝鮮総督府から『朝鮮巨樹老樹名木誌』が刊行され，朝鮮半島の3168 本の樹について同様のデータが記されている．本研究は，この二つの資料を基に，記載されている樹種や樹の所有者(所在地)，伝承を整理し比較した．樹種でいえば，掲載本数の多い順に『大日本老樹名木誌』ではマツ，スギ，クスノキ，ケヤキ，サクラ，イチョウと続き，『朝鮮巨樹老樹名木誌』ではケヤキ，エノキ・ムクエノキ，イチョウ，チョウセンアカマツ(アカマツのこと)，ヤチダモ，エンジュと続く．樹の所有は日本では，神社や寺院に植えられている割合が高い．一方，朝鮮半島では公有地，地域共同体である「里」や「洞」が圧倒的に多い．これは，地域で樹を祭る習慣があるためである．樹にまつわる伝承をその内容に基づいて分類した結果，樹種による違いが明確に表れた．それぞれの樹種が有する形態的，生理的特徴に起因すると考えられるものも多い．また，日本と朝鮮半島を比較すると，共通点もみられるが，むしろ異なる部分が目立ち，歴史，宗教，文化の違いが反映されている．


Triclosan, an antibacterial agent, increases intracellular Zn2+ concentration in rat thymocytes: Its relation to oxidative stress

Triclosan is used as an antibacterial agent in household items and personal care products. Since this compound is found in maternal milk of humans and bodies of wild animals, there is growing concern among some consumer groups and scientific community that triclosan is adverse for humans and wild animals. In order to estimate adverse actions of triclosan, the effects of triclosan on intracellular Zn2+ concentration and cellular thiol content were studied in rat thymocytes by the use of flow cytometer with appropriate fluorescent probes. Triclosan at 1-3 µM (sublethal concentrations) increased the intensity of FluoZin-3 fluorescence (intracellular Zn2+ concentration) and decreased the intensity of 5-chloromethylfluorescein (5-CMF) fluorescence (cellular thiol content). Negative correlation (r = -0.985) between triclosan-induced changes in FluoZin-3 and 5-CMF fluorescences was found. Removal of external Zn2+ did not significantly affect the triclosan-induced augmentation of FluoZin-3 fluorescence, suggesting an intracellular Zn2+ release by triclosan. These actions of triclosan were similar to those of H2O2 and triclosan significantly potentiated the cytotoxicity of H2O2. Therefore, the results may suggest that triclosan at sublethal concentrations induces oxidative stress that decreases cellular thiol content, resulting in an increase in intracellular Zn2+ concentration by Zn2+ release from intracellular store(s). Since recent studies show many physiological roles of intracellular Zn2+ in cellular functions, the triclosan-induced disturbance of cellular Zn2+ homeostasis may induce adverse actions on the cells.


Elevation of intracellular Ca2+ level by triclosan in rat thymic lymphocytes: Increase in membrane Ca2+ permeability and induction of intracellular Ca2+ release

Triclosan is an antibacterial agent used in household items and personal care products. Because wild animals and humans can harbor this compound in their systems, the toxic effects of triclosan are a possibility and are suspected. Therefore, we examined the effects of triclosan on intracellular Ca2+ concentration in rat thymocytes by cytometric techniques using fluorescent probes. Triclosan doses of 1 10 µM significantly increased the intensity of Ca2+-detecting Fluo-3 fluorescence, indicating an increase in intracellular Ca2+ concentrations. The augmentation of Fluo-3 fluorescence became more profound in a dose-dependent manner after the addition of an external source of Ca2+. Conversely, the removal of external Ca2+ greatly attenuated the triclosan-induced augmentation of Fluo-3 fluorescence. These results suggest that triclosan treatment allows external Ca2+ to pass through cell membranes. This phenomenon was not specific for Ca2+ because external Mn2+ quenched the triclosan-induced augmentation of Fluo-3 fluorescence, indicating that triclosan can also mediate Mn2+ permeation across membranes. Therefore, these results suggest that triclosan increases membrane permeability to divalent metal cations. Furthermore, triclosan induces Ca2+ release from intracellular stores because the Fluo-3 fluorescence intensity still increased slightly after triclosan treatment, even under conditions free from external Ca2+. Additionally, triclosan did not increase the intensity of Fluo-3 fluorescence when Ca2+ was depleted from intracellular Ca2+ stores by A23187 under the external Ca2+-free condition. Taken together, these data suggest that micromolar concentrations of triclosan affect intracellular Ca2+ homeostasis in thymocytes, possibly resulting in cellular malfunction.


Some characteristics of membrane Cd2+ transport in rat thymocytes: An analysis using Fluo-3

Although cadmium-induced apoptosis of lymphocytes is one of common features in the immunotoxicity of cadmium, the membrane pathway for intracellular cadmium accumulation is not fully elucidated. To characterize membrane Cd2+ transport of rat thymocytes, the change in intracellular Cd2+ concentration under various conditions was examined by the use of Fluo-3, a fluorescent probe for monitoring the change in intracellular concentration of divalent metal cations. The membrane Cd2+ transport was estimated by the augmentation of Fluo-3 fluorescence induced by bath application of CdCl2. Lowering temperature strongly suppressed the augmentation of Fluo-3 fluorescence by CdCl2, suggesting that the metabolic process can be involved in membrane Cd2+ transport. External acidification (decreasing pH) and membrane depolarization by adding KCl attenuated the augmentation, indicating the requirement of electrochemical driving force for membrane Cd2+ transport into the cells. Bath application of CaCl2 and ZnCl2 equally decreased the augmentation, suggesting their competition with Cd2+ at the membrane transport. The augmentation by CdCl2 was lesser in the cells treated with N-ethylmaleimide inducing chemical depletion of cellular thiols. The result suggests the contribution of sulfhydryl groups to membrane Cd2+ transport. Taken together, it is suggested that the cells possess a temperature-sensitive membrane Cd2+ pathway, driven by electrochemical gradient of Cd2+ and transmembrane potential, with competitive binding site. Based on the characteristics described above, it is unlikely that the membrane Cd2+ transport in rat thymocytes is attributed to a single transport system although it has characteristics that are similar to those of divalent cation transporter 1.


Putative role of intracellular Zn2+ release during oxidative stress: A trigger to restore cellular thiol content that is decreased by oxidative stress

Although the ability of zinc to retard oxidative process has been recognized for many years, zinc itself has been reported to induce oxidative stress. In order to give some insights to elucidate the role of intracellular Zn2+ in cells suffering from oxidative stress, the effects of N-ethylmaleimide (NEM) and ZnCl2 on cellular thiol content and intracellular Zn2+ concentration were studied by use of 5-chloromethylfluorescein diacetate (5-CMF-DA) and FluoZin-3 pentaacetoxymethyl ester (FluoZin-3-AM) in rat thymocytes. The treatment of cells with NEM attenuated 5-CMF fluorescence and augmented FluoZin-3 fluorescence in a dose-dependent manner. These NEM-induced phenomena were observed under external Zn2+-free condition. Results suggest that NEM decreases cellular thiol content and induces intracellular Zn2+ release. Micromolar ZnCl2 dose-dependently augmented both FluoZin-3 and 5-CMF fluorescences, suggesting that the elevation of intracellular Zn2+ concentration increases cellular thiol content. Taken together, it is hypothesized that intracellular Zn2+ release during oxidative stress is a trigger to restore cellular thiol content that is decreased by oxidative stress.


Methylmercury elicits intracellular Zn2+ release in rat thymocytes: Its relation to methylmercury-induced decrease in cellular thiol content

We have previously revealed that thimerosal, an organomercurial preservative, increases intracellular Zn2+ concentration in rat thymocytes. Because thimerosal contains ethylmercury that confers the toxicity, it is a possibility that methylmercury (MetHg), an environmental pollutant, also increases intracellular Zn2+ concentration. This possibility was tested by measuring intracellular Zn2+ level with FluoZin-3, a fluorescent probe for intracellular Zn2+. MetHg at concentrations ranging from 100 nM to 1 µM significantly increased the intensity of FluoZin-3 fluorescence, an indicator for intracellular Zn2+ concentration, under external Ca2+- and Zn2+-free condition in a concentration-dependent manner. TPEN, a chelator for intracellular Zn2+, completely diminished the MetHg-induced augmentation of FluoZin-3 fluorescence. MetHg at 100 nM or more significantly decreased the intensity of 5-chlormethylfluorescein fluorescence, an indicator for cellular thiol content. Such MetHg-induced changes in the fluorescence were correlated with a coefficient of -0.917. Taken together, it is suggested that submicromolar MetHg releases Zn2+ from intracellular thiol, resulting in the increase in intracellular Zn2+ concentration. However, it is unlikely that MetHg at critical maternal blood concentration (27 nM) affects intracellular Zn2+ homeostasis.


Tri-n-butyltin increases intracellular Zn2+ concentration by decreasing cellular thiol content in rat thymocytes

Effect of tri-n-butyltin (TBT), an environmental pollutant, on intracellular Zn2+ concentration was tested in rat thymocytes to reveal one of cytotoxic profiles of TBT at nanomolar concentrations using a flow cytometer and appropriate fluorescent probes. TBT at concentrations of 30 nM or more (up to 300 nM) significantly increased the intensity of FluoZin-3 fluorescence, an indicator for intracellular Zn2+ concentration, under external Ca2+- and Zn2+-free condition. Chelating intracellular Zn2+ completely attenuated the TBT-induced augmentation of FluoZin-3 fluorescence. Result suggests that nanomolar TBT releases Zn2+ from intracellular store site. Oxidative stress induced by hydrogen peroxide also increased the FluoZin-3 fluorescence intensity. The effects of TBT and hydrogen peroxide on the fluorescence were additive. TBT-induced changes in the fluorescence of FluoZin-3 and 5-chloromethylfluorescein, an indicator for cellular thiol content, were correlated with a coefficient of -0.962. Result suggests that the intracellular Zn2+ release by TBT is associated with TBT-induced reduction of cellular thiol content. However, chelating intracellular Zn2+ potentiated the cytotoxicity of TBT. Therefore, the TBT-induced increase in intracellular Zn2+ concentration may be a type of stress responses to protect the cells.


Mercury-induced oxidative stress in marine phytoplankton Tetraselmis tetrathele (Prasinophyceae).

Mercury-induced oxidative stress was observed in the cells of marine phytoplankton Tetraselmis tetrathele (Prasinophyceae) using the fluorescent probes 2' 7'-dichlorofluorescin diacetate (DCFH-DA) and fluorescein diacetate (FDA). When the cells were exposed to 1 μM of HgCl2, the fluorescence intensity of 2' 7'-dichlorofluorescin (DCF) was three times higher than that of the control level 60 min after the treatment, indicating H2O2 accumulation, and it gradually decreased. Moreover, exposure to 0.3 μM of HgCl2 caused an increase in fluorescence 6 h after the treatment. However, exposure to HgCl2 at concentrations of 3.0 μM or greater did not cause an increase in fluorescence. It is thought that H2O2 was also generated at these concentrations, but could not be detected due to inactivation of the cellular esterase necessary to convert DCFH-DA to DCFH, and/or leakage from the cells due to membrane damage caused by HgCl2. The maximum level of fluorescence was approximately two times higher under light conditions than under dark conditions, suggesting that H2O2 was derived partly from photosynthesis. These results indicate that accumulation of H2O2 is involved inconsequence of the toxicity of mercury.

広く海洋に生息し，海洋生態系の第一次生産者である植物プランクトン であるプラシノ藻類 Tetraselmis tetrathele を研究材料としてHg処理後の細胞の変化について，特にH2O2の発生に注目して調べた． T. tetratheleを様々な濃度のHgCl2で処理し，DCFH-DAで染色したところ，1時間及び3時間Hg処理において，1 μMのHgCl2濃度のときに強い蛍光がみられた．また6時間処理した細胞でも0.3 μMのHgCl2濃度で同様の強い蛍光がみられた．これは，細胞がHgの影響を受け，細胞内にH2O2が発生していることを示している． また，明条件下あるいは暗条件下において，1 μMのHgCl2で処理した細胞のDCF蛍光強度を時間を追って調べると，明条件では，60分から70分で最大の蛍光を示したが，暗条件では50分で最大の蛍光を示した．このことから，暗条件下では明条件下よりHgの影響をより早く受けることがわかった． 次にFDA染色により明条件と暗条件で細胞内エステラーゼ活性を比較した．その結果，共に1 μMのHgCl2において，酵素活性が著しく低下していることがわかった．また，明条件よりも，暗条件のほうが低い濃度のHgで酵素活性が低下した．

Natural Science Research, Faculty of Integrated Arts and Sciences, The University of Tokushima


The 88-temple pilgrimage in Shikoku Island and the nature around the pilgrimage route: histories of sacred trees, landscape and crests. --- 神木，自然景観，寺紋について

四国霊場と自然 --- 神木，自然景観，寺紋について

阿讃山地沿いに分布する四国霊場(1番札所霊山寺から5番札所地蔵寺)および大麻比古神社において，境内や周辺の自然景観の変遷，遍路道の変遷，そして神木の歴史，寺紋について調査を実施した．

徳島大学総合科学部自然科学研究


Tri-n-butyltin-induced blockade of store-operated calcium influx in rat thymocytes

Tri-n-butyltin (TBT), one of environmental pollutants, disturbs intracellular Ca2+ homeostasis by increasing intracellular Ca2+ concentration ([Ca2+]i). Effect of TBT on oscillatory change in [Ca2+]i (Ca2+ oscillation) of rat thymocytes was examined using a laser microscope with fluo-3-AM in order to further elucidate the TBT toxicity related to intracellular Ca2+. The Ca2+ oscillation was completely attenuated by 300 nM TBT. Since store-operated Ca2+ channels are involved in the generation of Ca2+ oscillation, the action of TBT on an increase in [Ca2+]i by Ca2+ influx through store-operated Ca2+ channels was examined. The increase in [Ca2+]i by the store-operated Ca2+ influx was not affected by 3 nM TBT. However, TBT at 10 nM or more significantly reduced the increase in [Ca2+]i. It is likely that TBT attenuates the Ca2+ oscillation by reducing the Ca2+ influx through store-operated Ca2+ channels.

Environmental Toxicology and Pharmacology


Increase in intracellular Cd2+ concentration of rat cerebellar granule neurons incubated with cadmium chloride: Cadmium cytotoxicity under external Ca2+-free condition

In order to examine the cadmium cytotoxicity unrelated to external Ca(2+), the effects of micromolar CdCl(2) on intracellular Cd(2+) concentration, cellular content of glutathione, and cell viability of rat cerebellar granule neurons were examined under normal Ca(2+) and external Ca(2+)-free conditions, using a laser confocal microscope with fluorescent probes, fluo-3-AM, 5-chloromethylfluorescein (CMF) diacetate, and propidium iodide. CdCl(2) (10-300muM) dose-dependently increased the intensity of fluo-3 fluorescence. Exposure to CdCl(2) equally enhanced the fluo-3 fluorescence under both Ca(2+) conditions and MnCl(2) did not quench the CdCl(2)-enhanced fluorescence. The results indicate that the enhancement of fluo-3 fluorescence is due to the increase in intracellular Cd(2+) concentration. CdCl(2) at 100-300muM decreased the intensity of CMF fluorescence, indicating the decrease in cellular content of glutathione. The population of cells stained with propidium (dead cells) was increased by 100-300muM CdCl(2). Similar results described above were also observed under external Ca(2+)-free condition. It is suggested that some of cytotoxic actions of CdCl(2) on neurons are unrelated to external Ca(2+), one of main sources for increasing intracellular Ca(2+) concentration.

カドミウム毒性はカルシウムイオンの細胞機能の阻害との関連で示唆されることが多い．そこでカルシウムイオン除去条件下で，カドミウムイオンそれ自体が細胞レベルで同様な細胞毒性を示すことを明らかにした．細胞内でカドミウムイオン濃度の上昇を画像化して，カルシウムイオンの有無に関わらず，カドミウムイオンは同程度の細胞毒性を示した．


Increase in number of annexin V-positive living cells of rat thymocytes by intracellular Pb2+

鉛は世界各地で健康被害を起こしている代表的な環境汚染物質の一つである．しかしながら，鉛の細胞レベルでの作用は多様であり研究が続いている．この論文では細胞内に入った鉛が細胞にアポトーシス様膜変化を起こすことを詳細に検討した．現在，米国でも90万人程度は血中鉛濃度がかなり高いことが報告されている．これに近い濃度で細胞内に入った鉛が細胞死のトリガーになる可能性を示した．


Cytotoxic effects of triphenylbismuth on rat thymocytes: Comparison with bismuth chloride and triphenyltin chloride

近年，有用性が増しているトリフェニルビスマスの環境毒性を考察する上で必要な細胞レベルでの毒性を検討した．環境汚染物質のトリブチルスズと比べると毒性は低いものの，μMのオーダーでは明らかに毒性が確認された．よって，その使用に於いては環境(自然環境，研究室環境)への配慮が必要である．


Determination of cellular levels of nonprotein thiols and their correlations with susceptibility to mercury in phytoplankton

We determined the intracellular contents and concentrations of cysteine and glutathione in five species of marine phytoplankton and examined relationships to mercury susceptibility. Intracellular concentrations of nonprotein thiols in mM level in the five species. Flow cytometric analyses with the dye, fluorescein diacetate (FDA), was used to monitor the effect of HgCl2 on esterase, and the 50%-effect concentrations (EC50s) were compared in the five species. The EC50s after 3 h exposure to HgCl2 correlated well with the GSH concentrations.

海洋性の植物プランクトンにおいて，解毒物質であるグルタチオンとシステインの定量を行い，細胞あたりの含量と細胞中の濃度を計算した．その結果，これらの物質がmMレベルで存在することが分かり，また細胞中濃度の高い種ほど酸化度が低いことも明らかになった． また，FDA染色法により，各植物プランクトンの水銀耐性を測定し，グルタチオンやシステインの含量との関係を調べた結果，グルタチオンやシステイン濃度の高い種ほど水銀に抵抗力を持つことが明らかになった．これは異なる種を用いてこれらの物質の含量と重金属抵抗性の相関を明らかにした初めての報告である．


Tri-n-butyltin-induced decrease in cellular level of glutathione in rat thymocytes: a flow cytometric study using 5-chloromethylfluorescein diacetate


Toxicity of methylmercury conjugated with L-cysteine on rat thymocytes in comparison with that of methylmercury


Flow cytometric analysis on tri-n-butyltin-induced increase in annexin V binding to membranes of rat thymocytes


Heavy-metal induced changes in cellular nonproteinaceous thiol levels and heavy metal-binding peptides in Tetraselmis tetrathele (Prasinophyceae)

The effects of mercury and cadmium on the intracellular level of nonprotein thiols in a unicellular green alga Tetraselmis tetrathele (Prasinophyceae) were investigated by using a fluorescent dye, 5-chloromethylfluorescein (5CMF), as a probe for nonprotein thiols. Depletion of nonprotein thiols began within 30 min, and completely depleted at 2 h, and most cells died at 24 h after exposure the cells to more than 3.0 M HgCl2. HPLC analyses revealed that glutathione was a major nonprotein thiol in T. tetrathele. A tripeptide, Arg-Arg-Glu, was found to be abundant and it showed ability to bind Hg2+, suggesting that it functions to scavenge heavy metals as well as thiol molecules.


Spermine is a salicylate-independent endogenous inducer for both tabacco acidid pathogenesis-related proteins and resistance against tobacco mosaic virus infection

タバコモザイクウィルスを感染させたタバコ葉の病斑部位の細胞間隙に，スペルミンが健康葉の20倍も蓄積していることを見いだした．そして，健康葉にスペルミンを投与することにより，PR protein の発現が誘導されること，およびタバコモザイクウィルスによる病斑が縮小することが分かった．このスペルミンによる防御メカニズムは，防御反応誘導物質であるサリチル酸とまったく独立に働く新規のメカニズムであることが明らかになった．


Methylmercury toxicity in rat brain neurons: modificatons by compounds possessing SH-group and comparison with dimethylmercury, N-ethylmaleimide, and ionomycin

Environmental Toxicology and Pharmacology


Effects of triphenyltin on growth and viability of K562 leukemia cells


Isolation of organellar DNA from Codium fragile (Codiaceae, Codiales, Ulvophyceae)

緑藻類ミルCodium fragile から塩化セシウム密度勾配超遠心によりオルガネラDNAを単離した．超遠心の結果，三本のバンドが存在し，サザン法により核DNA，葉緑体DNA，ミトコンドリアDNAを同定した．また核のrRNAはミトコンドリアDNAと同じ位置にバンドを形成した．ミトコンドリアDNAのゲノムサイズは141.6 kb以上であり，微細藻類より大きく，陸上植物より小さい値であった．


Isolation of eyespots of green algae and analyses of pigments

遊走性を示す植物プランクトンの多くは，眼点と呼ばれる細胞内小器官を有し，走光性を示す．本研究は緑色藻類を用いて眼点の単離·精製法を確立し，眼点の構造と眼点中の色素組成を調べた．その結果，Pyramimonasではβ-カロテンが，Mesostigmaではγ-カロテンが，Bryopsisではα-カロテンが主成分であった．これらの成分の違いは，走光性のメカニズムとは関係なく，カロテンの代謝メカニズムによると推察した．


Organization of heterogeneous mitochondrial DNA molecules in mitochondrial nuclei of cultured tobacco cells

高等植物のミトコンドリアは多数のDNA分子種が存在し，個々のミトコンドリアにおいてどのように統合されているのか問題となっていた．そこで，タバコ培養細胞において，ミトコンドリアに含まれるDNA量，ミトコンドリアゲノムサイズ，ミトコンドリアDNA分子種の長さについて調べた．その結果，個々のミトコンドリアは異なるDNA分子種を複数もつが，総ての分子種を持つわけではないこと，すなわちゲノム構成が異なるミトコンドリアが存在することを証明した．


Inter- and intraspecific variations of chloroplast DNA of the siphonous green algal genus Caulerpa (Caulerpales, Chlorophyta)

緑藻類イワヅタ葉緑体DNAの精製法を確立し，日本産のイワヅタ4種についてその制限酵素パターンを比較した．その結果，大きな種間変異がみられたが，外部形態や葉緑体の構造で共通特徴を持つ2種については制限酵素パターンに類似性がみられた．また，この2種については葉緑体DNAのゲノムサイズが高等植物に比べて著しく小さいことが明らかになった．


Organization of multiple nucleoids and DNA molecules in mitochondria of human cell

ヒト卵巣癌由来A2780培養細胞におけるミトコンドリアDNAおよびミトコンドリア核の存在様式を明らかにした．すなわち，ミトコンドリアあたり平均4.6コピーのミトコンドリアDNAが存在すること，細胞あたり約500コピーのミトコンドリアDNAが存在することを明らかにした．また，ミトコンドリアのうち70%は複数のミトコンドリア核を持つ''多核''であることを初めて明らかにした．


Present situation of Elaeocarpus zolloingeri tree planted in Mt. Shiroyama in 2006 --- survival and infection status of Elaeocarpus yellows (Second Report)

2006年に徳島市城山に植樹されたホルトノキの苗木の現状 --- 生育状況とホルトノキ萎黄病感染状況について (第二報)

This paper is an additional report of the 2016 survey report on the saplings of Elaeocarpus zollingeri (synonym: E. sylvestris) in Mt. Shiroyama in the Tokushima City. Since 1970s community of E. zollingeri in Mt. Shiroyama has continued to decline by Elaeocarpus yellows caused by phytoplasma. To prevent extinction 300 saplings were planted at the foot of the mountain by volunteers in 2006. In 2016, 10 years after planting, 40 trees had been alive, and the number has decreased to 29 in 2022. There were two other saplings near the planted area, presumably naturally occurring seedlings. The most grown tree was over 5 m in height, while there were 4 trees less than 1 m. These saplings were checked for phytoplasma infection by nested PCR, and 9 saplings were found to be infected. However, none of them shows symptoms of the disease in appearance, and there is no difference in tree height compared to the uninfected saplings. To prevent further loss in the future, poorly growing saplings should be transplanted into better conditions. Also, in areas with many surviving saplings, it is advisable to transplant several saplings to avoid overcrowding.

ホルトノキ萎黄病により消滅の危機にある城山のホルトノキ群落を再生するため，城山のホルトノキの消滅を防ぐために，2006年にNPOらによって300本のホルトノキの苗木が城山山麓に植えられた．本研究はその追跡調査を行った．2022年の調査では300本の苗木のうち29本が生存していた．樹高は最も高い個体が5.2 m，最も低い個体が82 cm，平均は194 cmであった．また，これら29本および周辺の稚樹2本についてファイトプラズマに感染しているかどうかをnested PCRにより確認した結果，9本の感染が判明した．これ以上の消失を防ぐため，移植や新たな苗木の育成などの対策が急務である．

Regional Science Research, the University of Tokushima


Moraes's Garden --- (6)Moraes's Eyes: Tokushima Landscape through his Eyes

モラエスの庭 --- (6)モラエスの目:徳島の風景

This paper is part of the results from the social action activities financed by the Faculty of Integrated Arts and Sciences, and Tokushima University, 2015. The activities are mainly focused on the reevaluation by public lectures and other exhibitions of Wenceslau de Moraes 1854-1929, a Portugal naval officer and consul general who lived and died in Tokushima. This is also part of the outcomes of the Project Studies by Moraes's Studies Group launched on July 31, 2010.The members of Moraes's Studies Group, T. Miyazaki (English Literature, Comparative Literature), E. Ishikawa (German Literature, Comparative Literature), M. Satoh (Plant Physiology), M. Sakai (Clinical Psychology), all at the Institute of Socio-Arts and Sciences, Tokushima University, have been continuing to try to analyze Moraes's works and to approach new facets of Moraes's biographical aspects. Moraes was fascinated by the far-east Japan and fell in love with Ó-Yoné, who died soon after the marriage. After her death, Moraes decided to live in Tokushima, which was Ó-Yoné's hometown. He lived with Ko-Haru, Ó-Yoné's niece, for a while until she died from tuberculosis at the age of 21. His life until his death in Tokushima was a kind of a hermit, disregard of his fame as Consul General and Navy high-rank Officer of Portugal, and other financial merits entailed with them. Moraes published O ‟Bon-odori„ em Tokushima in 1916 after Ó-Yoné died, and Ó-Yoné é Ko-haru afterwards. In both works Moraes depicted Tokushima landscape and everyday things in general which his eyes caught through his keen sensitivity. He worshipped Japan's unique beautiful landscape, but he was disappointed at the rapidly westernized aspects of large cities like Kobe, Osaka and Tokyo. He had been, in a sense, allured and attracted by the old and beautiful landscape through the writings by his foregoing visitors to Japan. His choice of Tokushima as his residence for his remaining days might be considered to search for the old and traditional beautiful aspects of Japan that seemed to him to be remaining in a local city like Tokushima far away from the large cities. Moraes's sense of beauty through his eyes to Tokushima landscape may lead us Japanese to consider our identity in this now globalized society.


Moraes's Garden --- (5) Writings in the Time and the 5th National Industrial Exhibition

モラエスの庭 --- (5) モラエスの著作の位置づけと第五回内国勧業博覧会

This paper and report is based on the oral presentation delivered on 29th, November 2014, as part of Symposium `Moraes in Kobe and Tokushima' held at the Faculty of Integrated Arts and Sciences, Tokushima University, in Tokushima. This is also part of the outcomes of the Project Studies by the activities in 2014 of Moraes's Studies Group launched on July 31, 2010. The members of Moraes's Studies Group, T. Miyazaki (English Literature, Comparative Literature), E. Ishikawa (German Literature, Comparative Literature), M. Satoh (Plant Physiology), M. Sakai (Clinical Psychology), all at the Institute of Socio-Arts and Sciences, Tokushima University, have been continuing to try to analyze Moraes's works and to explore new facets of Moraes's biographical facts. Moraes was fascinated by the far-east Japan and fell in love with Ó-Yoné, who died young at the age of 39 in Kobe. After her death, Moraes decided to leave Kobe and live in Tokushima, which was Ó-Yoné's hometown. He lived with Ko-Haru, Ó-Yoné's niece, for a while until she died from tuberculosis at the age of 21. He led his life until his death in Tokushima for 16 years as a kind of hermit, neglecting his fame as the Consul General and Navy high-rank Officer of Portugal, and other financial merits entailed with the post. Though Moraes is often regarded as a kind of hermit in Tokushima, we should pay more attention to and reevaluate his aspect of a diplomat and consul in Kobe days before coming to Tokushima. He was a very able diplomat and consul, and once actively involved with the 5th National Industrial Exhibition held in 1903. In Tokushima he wrote and published O Bon-odori em Tokushima and afterwards Ó-Yoné e Ko-haru. These works might be regarded as based on the forms of diary and essay, seemingly as reports from Tokushima to Bento Carqueja, editor of Comércio do Porto (Porto Commercial Newspaper) in Portugal. Concerning these works as such there seems to be an undercurrent of the image of garden, or paradise, which is strongly connected with the Exhibition he was concerned. In this paper, a tentative reevaluation of his works in Tokushima and the undercurrent of the image of garden related with the 5th National Industrial Exhibition he was involved with in Kobe days.


Moraes's Garden --- (4) Looking to Life, and Looking to Death

モラエスの庭 --- (4)生へのまなざし，死へのまなざし

This paper is an essay on Moraes's O Bon-odori em Tokushima and Ó-Yoné e Ko-haru, part of the outcomes of the Project Studies by the activities in 2013 of Moraes's Studies Group launched in July 31, 2010. The members of Moraes's Studies Group, T. Miyazaki (English Literature, Comparative Literature), E. Ishikawa (German Literature, Comparative Literature), M. Satoh (Plant Physiology), M. Sakai (Clinical Psychology), all at the Institute of Socio-Arts and Sciences, The University of Tokushima, have been continuing to try to analyze Moraes's works and to approach new facets of Moraes's biographical aspects. Moraes was fascinated by the far-east Japan, and fell in love with Ó-Yoné, who died soon after the marriage. After her death Moraes decided to live in Tokushima, which was Ó-Yoné's hometown. He lived with Ko-Haru, Ó-Yoné's niece, for a while until she died from tuberculosis at the age of 21. His life until his death in Tokushima was a kind of hermit, disregard of his fame as Consul General and Navy high-rank Officer of Portugal, and other financial merits entailed with them. Moraes published O Bon-odori em Tokushima in 1916 after Ó-Yoné died, and Ó-Yoné e Ko-haru afterwards. This work might be regarded as based on the forms of diary and essay, seemingly as reports from Tokushima to Bento Carqueja, editor of Comércio do Porto (Porto Commercial Newspaper) in Portugal. He consistently wrote these installment reports from Tokushima in the eyes of a stranger, putting some distance between him and the people in there. Everything seen in the eyes of Moraes wore some beautiful visional aspect because of his memory of Ó-Yoné. He expressed his thoughts on life and death throughout O Bon-odori em Tokushima and Ó-Yoné e Ko-haru with fragmentary memories of his own as objective correlatives for the readers of his writings. This paper is based on the presentation in the Symposium at the 49th Annual Conference of Japan Comparative Literature Association Kansai Branch held at the Faculty of Integrated Arts and Sciences, here in Tokushima.


Moraess tomb, funeral, and rooms reported in the January 1930 issue of the Portuguese popular magazine CIVILIZAÇÃO

ポルトガルの大衆紙"CIVILIZAÇÃO''が1930年1月号で伝えたモラエスの墓， 告別式，彼の部屋に関する記事について

Portuguese popular magazine ``CIVILIZAÇÃO'' published an article of the tomb, funeral, and rooms of Wenceslau de Moraes in the January 1930 issue after about half a year when he died at his house in Tokushima on July 1, 1929. This article has not been introduced to Japan. Even though it is short, we can understand high evaluation of him in Portugal. Furthermore, it contains seven photos; four of them have not been published in any Japanese documents. These photos, three are of his rooms and one is taken at the funeral, tell us Moraes's life vividly with new findings about his belongings and acquaintanceship.

1929年7月1にヴェンセスラウ・デ・モラエスが亡くなってからおよそ半年後，ポルトガルの月刊大衆誌``CIVILIZAÇÃO'' が彼の墓，葬儀，部屋の様子を紹介する記事を1930年1月号に掲載した．この記事は，日本ではこれまで紹介されたことはない．短い記事ではあるが，ポルトガルの大衆雑誌が彼の死をどのように伝えているか分かる興味深い資料である．さらに，この記事に掲載されている7枚の写真は，日本の資料では見ることのなかったものや，より鮮明に映っているものが含まれており，モラエスの生活を窺い知ることのできる貴重な資料である．本論文は，記事の文章や写真から読み取れる情報を整理し，モラエスに関する新たな知見を紹介した．また，今後明らかにすべき点についても述べた．


Moraes's Garden --- (3) In the Eyes of a Stranger

モラエスの庭 --- (3)異邦人のまなざし

This paper is an essay on Moraes's O Bon-odori em Tokushima, part of the outcomes of the Project Studies by the activities in 2012 of Moraes's Studies Group launched in July 31, 2010. The members of Moraes's Studies Group, T. Miyazaki (English Literature), M. Satoh (Plant Physiology), M. Sakai (Clinical Psychology), all at the Institute of Socio-Arts and Sciences, The University of Tokushima, have been continuing to try to analyze Moraes's works and to approach a new facet of Moraes's biographical aspects. Moraes was fascinated by the far-east Japan, and fell in love with Ó-Yoné, who died soon after the marriage. After her death Moraes decided to live in Tokushima, which was Ó-Yoné's hometown. He lived with Ko-Haru, Ó-Yoné's niece, for a while until she died from tuberculosis at the age of 23. His life until his death in Tokushima was a kind of hermit, disregard of his fame as Consul General and Navy high-rank Officer of Portugal, and other financial merits entailed with them. Moraes published O Bon-odori em Tokushima in 1916 after O-Yoné died. This work might be regarded as based on the forms of diary and essay, seemingly as reports from Tokushima to Bento Carqueja, editor of Comércio do Porto (Porto Commercial Newspaper) in Portugal. He consistently wrote these installment reports from Tokushima in the eyes of a stranger, putting some distance between him and the people in there. Everything seen in the eyes of Moraes wore some beautiful visional aspect because of his memory of Ó-Yoné. He expressed his distress at the attitudes of Tokushima people at some sections in this book; that is, he was seen as a`ke-tojin,'an alien. This discrepancy and distancing from the people among whom he lived as a hermit, he seemed to see the deep gap between him and the people he loved, leading to the pathetic outcry at the final part of his letters to Bento Carqueja, the editor. This tentative paper intends to open a new perspective in a rather fixed image of Moraes and studies about him.


Three Books of Moraess Picture Postcards Living Areas, Tourist Resorts, and Religious Piety

モラエスの三つの絵葉書書簡集 — 絵葉書書簡からみえるモラエスの生活圏，旅行，信仰について —

Portuguese writer Wenceslau de Moraes sent a large amount of picture postcards to Portugal from Japan. 609 of them were donated to Tokushima city in 1989, and they are stored in the Moraes Museum located at the summit of Mt. Bizan. They were published as Moraes no Ehagaki-shokan and Moraes Ehagaki-shu I - IV respectively in 1994 and 2004, in Japan. Independent from the collection in Tokushima, a book of picture postcard-collection entitled "Permanências e Errâncias no Japão" was published from Fundação Oriente in 2004 in Portugal. In this paper, we characterized these books and summarized the data of the picture postcards concerning his living areas and tourist resorts where he visited. He was outing vigorously to famous temples, shrines, and tourist resorts in holiday while he lived in Kobe. However, such opportunity extremely reduced after he moved to Tokushima when he started a cloistered life. Although Buddhism gave a significant impact on his religious piety, that is able to be understood from his writings, his feeling to Shinto has been obscured. However, some picture postcards revealed his attraction to Shinto. There is a Moraess photo that was taken at a waterfall when he lived in Kobe. The place is often explained as Nunobiki-no-taki (Kobe Nunobiki Waterfall), but we found that the it was the Tsutsumi-ga-taki at Arima, now it is called Tsuzumigataki Waterfall in the Tsuzumigataki Park in Arima, Kobe.

ヴェンセスラウ・デ・モラエス(Wenceslau de Moraes)は，親族や友人に頻繁に絵葉書や書簡を送った．これらのうち，ポルトガルから徳島市に寄贈されたモラエス館所蔵の609通は，1994年(平成6)には『モラエスの絵葉書書簡』として，2004年(平成16)には『モラエス絵葉書集I』~『〃 IV』として刊行された．また，ポルトガルのコレクターが集め，2004年に``Permanências e Errâncias no Japão''として刊行された420通の絵葉書集もある．本研究は，これらの中から風景絵葉書や各地の風物の絵葉書に注目し，モラエスの生活圏や旅行に関するデータをまとめ，神戸時代と徳島時代で比較した．その結果，モラエスは，神戸での外交官時代，休日に頻繁に各地に旅行に出かけていたが，徳島に移り住んでからは滅多に遠出をせず，墓参を日課とする隠遁生活を送ったことが確認できた．また，彼は頻繁に寺社に参詣に出かけており，絵葉書の文面から見て取れる日本の宗教に対する彼の本音を探ったところ，著作では触れられることのなかった神道への愛着を認めることができた． また，数少ないモラエスの写真の中に，神戸時代に撮影された彼が滝を背景にして写っている写真があり，従来それは「布引の滝」とされていた．しかし，絵葉書データを詳細に分析した結果，これは有馬の「皷ヶ瀧」(現 鼓ヶ滝)であることが判明した．


Moraes's Garden --- (2) Change in the Quality of 'Essay'

モラエスの庭 --- (2)「随筆」の変質

This paper is an essay on Moraes's Tokushima no Bon-odori, part of the outcomes of the Project Studies by the activities in 2011 of Moraes's Studies Group launched in July 31, 2010. The members of Moraes's Studies Group, T. Miyazaki (English Literature), M. Satoh (Plant Physiology), M. Sakai (Clinical Psychology), all at the Institute of Socio-Arts and Sciences, The University of Tokushima, have been continuing to try to analyze Moraes's works and to approach a new facet of Moraes's biographical aspects. Moraes was fascinated by the far-east Japan, and fell in love with Oyoné, who died soon after the marriage. After her death Moraes decided to live in Tokushima, which was Oyoné's hometown. He lived with Koharu, Oyoné's niece, for a while until she died from tuberculosis at the age of 23. His life until his death in Tokushima was a kind of hermit, disregard of his fame as Consul General and Navy high-rank Officer of Portugal, and other financial merits entailed with them. Moraes published Tokushima no Bon-odori in 1913 after Oyoné died. This work might be regarded as based on the forms of diary and essay, seemingly as reports from Tokushima to Bento Carqueja, editor of Porto Commercial Newspaper in Portugal. His interest in Kino Tsurayuki's Tosa-Nikki (Tosa Diary), which was written in the persona of a woman, seems to be the key to understand the modification in the quality of Tokushima no Bon-odori. Though this work was written as a diary and an essay in the persona of alien people to Tokushima, the tone of this work was quite changed at the final part of his letters to Bento Carqueja, the editor. This tentative paper intends to open a new perspective in a rather fixed image of Moraes and studies about him.


The Firefly Which Wenceslau De Moraes Saw

A Portuguese writer, Wenceslau de Moraes (1854-1929), who lived in Tokushima 1913-1929, wrote the essay ``Será Ó-Yoné... Será Ko-Haru?...'' in June, 1918. In this essay there is a scene in which Moraes heard some children singing an old song of firefly-catching popular among people here in Tokushima in those days. He heard and wrote down the word ``tané-mushi'' in the song, and he seemed to have thought this word meant a firefly. However, it must have been his misunderstanding of ``ta-no-mushi'' which is found in an old children's song handed down in Tokushima district. The word ``ta-no-mushi'' means an insect of a rice field, indicating a kind of firefly, Luciola lateralis. In the essay, Moraes wrote that just after hearing the children's song, he saw a firefly in front of his house that was located near Mt. Bizan, and the firefly helped him find the keyhole to open the door of his house by its illumination. Today four species of firefly are found to live in Mt. Bizan: Luciola cruciata, L. lateralis, Lucidina biplagiata and Pyrocoelia disciollis. According to the interviews with some of the elder people in this area, and taking into consideration of the habits of the four firefly species, L. cruciata seems to be the most probable species which Moraes saw in the essay.

ヴェンセスラウ・デ・モラエスが大正7年(1918)6月に書いた短編 ``Será Ó-Yoné... Será Ko-Haru?...'' (邦題「おヨネだろうか⋯⋯コハルだろうか⋯⋯」)は，徳島での自らの生活を描くとともに，亡くなった二人の女性おヨネとコハルを懐かしむ内容の随想である．この作品の中で，蛍狩りからの帰りと思われる子供たちが歌う「蛍こい」の歌詞が "Hotaru-ko, tané-mushi koi!"(ホタル コ タネムシ コイ!)と記されている．「タネムシ」の用例について徳島に伝わるわらべ歌や民謡を調べたところ，「タネムシ」は見つからず，「タノムシ(田の虫)」であることが分かった．さらに京都でも「田の虫」と歌われるものがあった．「タネムシ」はモラエスの聴き間違いであると考えられる．「田の虫」は最近は，使われなくなったがホタルの別称であり，ホタルの中でも止水を好み，水田，池，湿原に生息する田の代表的な昆虫の一つヘイケボタルを指す． 本作では，子供たちが歌う蛍の歌を聞いた後で，モラエスは自分の家の戸口で一頭のホタルに遭遇する．徳島市の眉山周辺には，ゲンジボタル，ヘイケボタル，オバボタル，オオマドボタルの4種が生息する．これらの習性や文献調査および古老の話を総合すると，モラエスが見た蛍はゲンジボタルであった可能性が高い．

Regional Science Research, the University of Tokushima


Moraes's Garden --- (1) On the Forms of Diary and Essay

モラエスの庭 --- (1)日記文学・随筆文学ということ


A practical training in environmental education for traversal thinking of a historical heritage of Tokushima,Daiju dam in Yoshino river --- a educational significance of environmental education for symbiosis

体験型ゼミ『徳島の文化遺産「吉野川第十堰」から学ぶ自然と人間の共生』の実施と共生環境教育としての意義 --- 現代GP「豊饒な吉野川を持続可能とする共生環境教育」の一環として

徳島大学総合科学部では，昨年度の試行ゼミに引き続き，今年度より現代的教育ニーズ取組支援プログラムの一環として，環境教育のための体験型ゼミ「豊饒な吉野川を持続可能とする共生環境教育」の授業を開始した．現在，持続可能な社会システムの構築は，地球レベルの課題となってきている．環境問題は，国家レベルの政策と共に，地域社会の果たす役割が大きく，今後はさらに特色ある地域の取り組みが求められることが予測されている．この体験ゼミでは，環境問題が時代を超えた総合的な問題であることを体得するために，徳島の文化遺産である吉野川第十堰，地域の天然記念物である巨樹イチョウの調査，さらに伝統文化としての藍染めや食文化体験を吉野川周辺の動植物生態調査と組み合わせた総合体験学習型とした．総合科学の必要性を体験的に学ぶためにも，このように，地域に根ざした総合体験型学習を取り入れた環境教育プログラムは，総合科学教育の主要な柱の一つとして体系化してゆく必要性があると考えられる．

Journal of University Education Research


History and Folklores of Two Giant Trees of Ginkgo, "Yagami-no-icho" and "Tenjin-no-icho", in Ishii Town

ライバルイチョウ ∼「矢神のイチョウ」と「天神のイチョウ」の歴史∼

徳島県石井町には，約900 m離れて幹の周囲が10 mを超えるイチョウの樹が2本あり，ともに県の天然記念物に指定されている．また，これら2本の樹はそれぞれ那須与一，菅原道真にゆかりの伝説を有している．これらのイチョウに関する資料を収集・整理し，樹の歴史や古写真による樹の姿の変遷，伝説の形成課程について調べた．その結果，大正期に撮影された天神のイチョウの写真は現在の姿とよく似ているが，「乳」と呼ばれる気根の多くは大正期以降に切断された形跡があること，イチョウと那須与一や菅原道真を結びつけた伝説の形成には，2つの神社の間の対抗意識が働いたように思われること，などが明らかになった．

Journal of Human Sciences and Arts Faculty of Integrated Arts and Sciences The University of Tokushima

徳島大学総合科学部人間社会文化研究


History of the Giant Camphor Tree at Yagami in Tokushima

徳島県藍住町春日神社の「矢上の大クス」と呼ばれる幹の周囲が約13 mのクスノキの巨樹の歴史について調べた．この樹は江戸時代の文化8年(1811)の『阿波名所図会』に描かれている有名な樹であるが，樹に関する資料は，単発的な調査によるものばかりであり，過去からの記録を整理しまとめてたものがこれまでなかった．そこで，史料を蒐集し読解したところ，大変貴重で面白い史料であってもこれまで引用されることなく埋もれてしまっているものがいくつも見つかったり，あるいは誤った内容が今日まで伝えられていることを見いだした．本樹については，上に挙げた江戸時代の絵図の他，大正時代にそれと同じ角度から撮影された写真が存在し，それらと現在の姿と比較することにより，一世紀毎，およそ200年間の樹の姿の変化が明らかになった．

徳島大学総合科学部人間社会文化研究


Comparative study on the primary structure of globin chains of the extracellular giant hemoglobin from the polychaet Perinereis aibuhitensis and Tylonrrhynchus heterochaetus

Natural Science Research, Faculty of Integrated Arts and Sciences, The University of Tokushima

徳島大学総合科学部自然科学研究


Nonprotein thiol levels and susceptibility to HgCl2 of harmful phytoplankton Heterocapsa circularisquama (Dinophyceae)

Nonprotein thiol levels of harmful marine phytoplankton Heterocapsa circulari-squama Horiguchi (Dinophyceae) were measured and compared to those of other phytoplankton. The concentrations of cysteine and GSH, reduced forms of nonprotein thiols, of H. circularisquama were 0.69 and 0.12 mM and were in a low range among the six phytoplankton previously reported. However, concentrations of cystine and GSSG, oxidized forms of nonprotein thiols, were 0.55 and 0.20 mM, and were not in a low level, and as a result, oxidation ratios of cysteine-cystine and GSH-GSSG were in a high range among the six phytoplankton. The oxidation ratios of nonprotein thiols in the phytoplankton were strongly correlated with the concentrations of reduced forms of nonprotein thiols, and the result confirmed the previous observation that the higher the concentration of the reduced form of nonprotein thiols the less they are oxidized. Susceptibility of H. circularisquama to HgCl2 was measured by a flow cytometric analysis using FDA staining, and H. circularisquama was found to be weak against HgCl2 compared to the other five phytoplankton perhaps due to its low concentrations of the nonprotein thiols. EC50s of 3 h exposure to HgCl2 in the six phytoplankton including H. circularisquama significantly correlated with the concentrations of cysteine and GSH. This result reinforced the previous observation that the intracellular concentrations of the nonprotein thiols reflect susceptibility to HgCl2.

二枚貝に対して毒性をもつ植物プランクトン Heterocapsa circularisquama について細胞内のシステインおよびグルタチオンを定量し，さらにFDA染色法により水銀感受性を測定した．その結果， H. circularisquama のシステインおよびグルタチオンの細胞内レベルは他のの植物プランクトンに比べて低く，酸化度は高かった．また，水銀に対して高い感受性を示した．よって先に報告したnonprotein thiol濃度の低い種ほど酸化度が高く，水銀に弱いという傾向がより強化された．

Natural Science Research, Faculty of Integrated Arts and Sciences, The University of Tokushima


Improved dehydroascobate assay using ascorbate oxidase

アスコルビン酸の定量法の1つとして，アスコルビン酸オキシダーゼを用いる方法がある．アスコルビン酸の酸化物であるデヒドロアスコルビン酸を測定する場合は，DTTによる還元反応と組み合わせる方法がとられるが，従来用いられている方法では，DTTとアスコルビン酸オキシダーゼが混在することにより，デヒドロアスコルビン酸を正確に測定できないことを見いだした．そこで，この方法の改良を試みた．そして，DTTの還元反応を行った後，水銀処理により未反応のDTTを不活性化することにより，この問題を解決し，デヒドロアスコルビン酸の正確な定量が可能となった．


Cytotoxic action of tri-n-butyltin hydride, a nucleaphilic triorganotin, on rat thymocytes: a flow cytometric study


Protective action of tri-n-butyltin on the cells suffering from oxidative strss: Its implication in bioaccumulation


Relationship between changes in biota and temprature of seawater at Ryuuguunoiso in Naruto

鳴門市竜宮の磯における生物相の変化と海水温度変化について

During 1996-2000 three species of green algae, Monostroma latissimum, Enteromorpha linza, and Ulva pertusa remarkably changed in both biomass and in periods of thallus stage at Ryuuguunoiso in Naruto City in Tokushima Prefecture. Large communities of M. latissimum and E. linza were found from December to spring in 1995-1996 and 1996-1997 seasons, but they could not be found in 1997-1998 and 1998-1999 seasons. In 2000 they were found again about two month later than before. Because temperatures of seawater at Naruto were higher from winter to spring in 1997-1998 and 1998-1999 seasons, when the thalli could not be found, than those in other three seasons, the high temperatures may have inhibited the growth of the two species. Large communities of U. pertusa were found from spring to summer in 1996, and communities became smaller and their existence period became shorter in 1997 and 1998 seasons, and it could not be found in 1999 and 2000 seasons. This disappearance corresponds to a tendency toward a rise of the mean temperatures of the seawater a year. In animals, a mollusk Aplysia kurodai showed a remarkable change in the day of the first observation of adult individuals in respective seasons. The first observation days were late in April in 1996 and 1997, and early in March in 1998, and late in January in 1999 and 2000, so it is about three months earlier in the last two years than the first two years. However, days of disappearance of adult individuals were not so different in the five years. Therefore, growth rate in an early life stage, when A. kurodai is small and can not be visible to eye observation, must be related to the change in the first observation days of the adults. The mean temperature of seawater in August-December was lower in 1996 and 1997 than those in other three years, and it was higher in 1999 and 2000 than other three years, suggesting that the high temperature in 1999 and 2000 promoted the growth of A. kurodai.

徳島県鳴門市竜宮の磯において個体数や出現時期が過去5年間に大きく変化した生物種を記載した．さらに海水温度との関係を調べた結果，緑藻類3種(ヒロハノヒトエグサ，ウスバアオノリ，アナアオサ)では高海水温度が成長を抑制し，軟体動物アメフラシでは，アメフラシでは逆に高海水温度が成長を促進する傾向がみられた．

徳島大学総合科学部自然科学研究


Effects of organic and inorganic mercuries on growth and viability of K562 human leukemia cells


Species variety and seasonal changes of macroalgae at Ryuuguunoiso in Tokushima

鳴門市竜宮の磯における大型藻類の種類と季節変化

鳴門市にある竜宮の磯における大型藻類の種類と季節変化について調べた．その結果，緑藻類は4目5科6属8種を，褐藻類は7目10科15属20種を，紅藻類は6目20科30属47種を確認した．淡路島や県南部地域，室戸岬の予備的な調査との比較では，各地域毎に種類の違いが見られ，また同じ種においても季節変化に若干の違いが見られた．

徳島大学総合科学部自然科学研究


Effects of N,N-dimethyldodecylamine-N-oxide on some cellular parameters of rat thymocytes

N,N-Dimethyldodecylamine-N-oxide (DDAO) is an amphoteric surfactant used in many detergents for kitchens. In this study, the effects of DDAO (10 100 µM) on cell lethality, intracellular Ca2+ level, intracellular Zn2+ level, and cellular content of nonprotein thiol were examined in mammalian cells (rat thymocytes) to further characterize its cytotoxicity. DDAO at the concentration of 100 µM (22.9 mg/L) slightly, but significantly, increased the parameters described above, and it showed no significant effect at the concentrations of 30 µM (6.87 mg/L) or less. Therefore, it is unlikely that DDAO at environmentally-relevant concentrations (< 10 70 ng/L) exerts toxic actions on wild mammals and humans.

N,N-Dimethyldodecylamine-N-oxide (DDAO) is an amphoteric surfactant used in many detergents for kitchens. In this study, the effects of DDAO (10–100 μM) on cell lethality, intracellular Ca2+ level, intracellular Zn2+ level, and cellular content of nonprotein thiol were examined in mammalian cells (rat thymocytes) to further characterize its cytotoxicity. DDAO at the concentration of 100 μM (22.9 mg/L) slightly, but significantly, increased the parameters described above, and it showed no significant effect at the concentrations of 30 μM (6.87 mg/L) or less. Therefore, it is unlikely that DDAO at environmentally-relevant concentrations (< 10–70 ng/L) exerts toxic actions on wild mammals and humans.


Nonivamide, a natural analog of capsaicin, affects intracellular Ca2+ level in rat thymic lymphocytes

Effect of nonivamide, a natural analog of capsaicin, on intracellular Ca2+ level of rat thymocytes was examined using a flow-cytometric technique with appropriate fluorescent probes in order to further characterize the cytotoxicity because nonivamide can be used as an active intergradient of antifouling paints. Nonivamide at concentrations ranging from 30 µM to 300 µM significantly increased the intensity of Fluo-3 fluorescence. The potency of 100 µM nonivamide to increase the fluorescence was similar to that of 100 µM capsaicin. The increase in Fluo-3 fluorescence by 100 µM nonivamide was attenuated under an external Ca2+-free condition. Nonivamide at 100 µM also increased the intensity of Fluo-3 fluorescence in the continued presence of 100 µM capsaicin. It is suggested that nonivamide at high micromolar concentrations increases intracellular Ca2+ level via the activation of vanilloid receptors. Nonivamide concentrations (30 µM or more) that increase intracellular Ca2+ level in rat thymocytes are comparable to those in algal cells. However, it is something hard to argue the implications in environmental science because nonivamide doesn't seem to be released into environment in such a high concentration, and because bioaccumulation of nonivamide has not been reported.