1.

論文

論文
矢倉, 公隆
出版情報: 平成2年科学研究費補助金 一般研究(C) 研究成果報告書 = 1990 Fiscal Year Final Research Report Summary.  1989-1990  pp.16p.-,  1991-03-01.  金沢大学教育学部
URL: http://hdl.handle.net/2297/46422
概要: 本研究は、植物の特定DNA配列がゲノム内でどのような分布をしているかを明らかにする事を目的とし、ソラマメ由来の高度反復配列であるBamHlファミリ-(250bp及び1500bp配列)あるいはタバコゲノムより単離した自己複製能をもった配列(A RS)をプロ-ブとしたサザン・ハイブリダイズ法及びin situハイブリダイズ法による解析を行い以下のような結果を得た。 1.ソラマメ(vicia faba)BamHl ファミリ-の250bpと1500bpの相同配列が、カラスノエンドウ(Vicia angustifolia)およびスズメノエンドウ(Vicia hirsuta)のソラマメ属の野生種にも分裂散型反復配列として存在することが明かとなり、さらに、250bp配列と1500bp配列は古くは互いに隣接した形でのみゲノム内に存在したいたものが、進化の過程で分離しながら別のファミリ-を形成するようになったということが示唆された。またカラスノエンドウ染色体における1500bp相同配列の分布に注目すると、ほとんどの場合、長腕あるいは短腕のテロルメもしくは動原体付近に見出され、長腕の中間の位置には存在しないか、存在していてもわずかであることが分かった。1500bp配列が全ての染色体の全長にわたって存在しているソラマメゲノムにおける分布様式とこの結果を対比させて考えると、この配列が進化と共に分散化が強まっていったことが示唆される。2.タバコ ARSと相同な配列はタバコ以外のいくつかの植物種にも存在することが分かった。その中で、ソラマメについてin situ ハイブリダイゼイションを行ったところ、明確な結果は未だ得られていないが、傾向として、銀粒子が各染色体に存在することを示唆するパタ-ンが多数得られた。今後、さらに比活性の高いプロ-ブを得るか、ARSと相同な配列をソラマメから単離しされをプロ-ブに用いることにより確かなデ-タを得る必要があろう。 The purpose of this research is to clarify distributional patterns of specific DNA sequences in plant genomes. The following results were obtained by Southern blot hybridization and in situ hybridization utilizing highly dispersed repeated sequences of Vicia faba BamHl families and a cloned autonomously replicating sequence (ARS) from tobacco nuclear genome as probes. 1. Sequences which have homology to 250bp and 1500bp BamHl family sequences of V. faba were also present as dispersed repeated sequences in genomes of two wild Vicia species, V. angustifolia and V. hirsuta. In addition, it was suggested that all ancester sequences of the 250bp and 1500bp BamHl families were arranged adjacently in the ancient time and some of them were split to generate other sequence families such as 250bp and 1500bp families in the process of evolution. In situ hybridization of the 1500bp sequence to 6pairs of subtelocentric chromosomes of V. angustifolia revealed that this sequence were localized in telomeric regions of short or long arms, or in the region near centromeres of each chromosome but not in the middle regions of long arms of any chromosomes. From the fact that the 1500bp sequence distributes diffusely throughout all the chromosomes of V. faba, it was also suggested that the extent of dispersion of this sequence was increased with the evolution of Vicia genome. 2. By Southern blot hybridization analysis it was found that sequences homologous to tobcco ARS were present in several other plant genomes. Although clear patterns were not obtained yet in situ hybrization using sequence homology to tobacco ARS as a probe showed that silver grains were seemed to be distributed every chromosomes of V. faba. In order to obtain fine results, it is necessary to use probe with higher specific activity or to clone ARS-homologows sequences from V. fava genome and use the probes.<br />The purpose of this research is to clarify distributional patterns of specific DNA sequences in plant genomes. The following results were obtained by Southern blot hybridization and in situ hybridization utilizing highly dispersed repeated sequences of Vicia faba BamHl families and a cloned autonomously replicating sequence (ARS) from tobacco nuclear genome as probes.1. Sequences which have homology to 250bp and 1500bp BamHl family sequences of V. faba were also present as dispersed repeated sequences in genomes of two wild Vicia species, V. angustifolia and V. hirsuta. In addition, it was suggested that all ancester sequences of the 250bp and 1500bp BamHl families were arranged adjacently in the ancient time and some of them were split to generate other sequence families such as 250bp and 1500bp families in the process of evolution. In situ hybridization of the 1500bp sequence to 6pairs of subtelocentric chromosomes of V. angustifolia revealed that this sequence were localized in telomeric regions of short or long arms, or in the region near centromeres of each chromosome but not in the middle regions of long arms of any chromosomes. From the fact that the 1500bp sequence distributes diffusely throughout all the chromosomes of V. faba, it was also suggested that the extent of dispersion of this sequence was increased with the evolution of Vicia genome.2. By Southern blot hybridization analysis it was found that sequences homologous to tobcco ARS were present in several other plant genomes. Although clear patterns were not obtained yet in situ hybrization using sequence homology to tobacco ARS as a probe showed that silver grains were seemed to be distributed every chromosomes of V. faba. In order to obtain fine results, it is necessary to use probe with higher specific activity or to clone ARS-homologows sequences from V. fava genome and use the probes.<br />研究期間(年度):1989-1990, 研究課題/領域番号:01540562 続きを見る
2.

論文

論文
服部, 憲尚
出版情報: 金沢大学十全医学会雑誌.  107  pp.174-182,  1998-04-20.  金沢大学十全医学会
URL: http://hdl.handle.net/2297/9305
概要: 金沢大学医学部医学科
3.

論文

論文
久慈, 一英
出版情報: 金沢大学十全医学会雑誌.  103  pp.252-264,  1994-04-20.  金沢大学十全医学会
URL: http://hdl.handle.net/2297/8530
4.

論文

論文
Kawahara, Ei ; Akasofu-Imai, Miwa ; Mizutani, Tomohiro ; Umei, Tamiko ; Kanitani, Tomokatsu ; Takeuchi, Yuzuru
出版情報: 金沢大学つるま保健学会誌 = Journal of the Tsuruma Health Science Society Kanazawa University.  33  pp.67-71,  2009-12-25.  金沢大学つるま保健学会 = the Tsuruma Health Science Society, Kanazawa University
URL: http://hdl.handle.net/2297/20311
概要: 高感度 in situ ハイブリゼーション(ISH)法と高感度 PCR 法を用いて子宮頸部扁平上皮病変と腺病変での16型と18型のヒト乳頭種ウイルス感染頻度検索し、核への組み込みの頻度を ISH 法で決定した。16, 18型は PCR と ISH でともに、扁平上皮系病変のグレードが高まるにつれて感染頻度が上がった。浸潤扁平上皮癌のすべての例で ISH で組み込み型を示し、CIN1 の全てでエピソーム型を示した。腺系病変では PCR でも ISH でも16型より18型が多く検出され、そのほぼすべてが組み込み型だった。CIN2ではエピソーム型が多くCIN3では組み込み型が多かった。これらから16, 18型の核への組み込みは扁平上皮病変でも腺病変でも癌の初期に現れる変化、あるいは癌への進展を強く示唆する所見と考えられた。Highly sensitive in situ hybridization (ISH) and highly sensitive PCR methods were used to determine the infection of HPV type 16 (HPV 16) and type 18 (HPV 18) and the integration of them into the cells of squamous and glandular lesions of the uterine cervix. The frequency of HPV infection detected by ISH and PCR increased with progression of squamous lesions. All cases of ICC showed the integrated pattern of positive signals with ISH and all positive cases in CIN1 showed the episomal pattern. In CIN2 episomal patterns were more frequent than integrated pattern and in CIN3 integrated pattern were more frequent. Glandular lesions showed a greater frequency of HPV 18 than HPV 16 on nested PCR and ISH, and almost all of ISH-positive cases showed the integrated pattern. These data suggested that integration of HPV 16 and 18 is required for carcinogenesis in both squamous cell carcinoma and adenocarcinoma of the uterine cervix. 続きを見る
5.

論文

論文
水上, 勇治 ; Mizukami, Yuji
出版情報: 平成11(1999)年度 科学研究費補助金 基盤研究(C) 研究成果報告書 = 1999 Fiscal Year Final Research Report Summary.  1998-1999  pp.14p.-,  2000-03.  金沢大学保健学系
URL: http://hdl.handle.net/2297/00049261
概要: 本研究ではmixed medullary-follicular carcinomaの組織発生を明らかにするために臨床病理学的,形態学的,生化学的検討を行った.結果は次の様であった.1.臨床病理学的には,本症は通常の髄様癌に比し,1).男性に 多い(6:1 vs 1:1).2).より若年者に多い(平均年齢36歳 vs 44歳).3).予后が良好である(5年生存率100% vs 78%).などの特徴がみられ,通常の髄様癌とは異なる臨床病理学的性格を示唆するものであった.2.髄様癌,mixed medullary-follicular carcinomaおよび種々の疾患の人甲状腺組織を用い,カルシトニン,サイログロブリンmRNA発現,蛋白発現をnorthern blot法,in situ hybridization法,免疫組織化学法を用い検討した.3.Northern blot法については甲状腺組織よりのRNA抽出に不備があり,このためカルシトニン,サイログロブリンmRNAの定量的観察が出来なかった.しかしin situ hybridization法,免役組織化学法についてはほぼ一致した結果が得られた.4.In situ hybridization法,免役組織化学法ともに,mixed medullary-follicular carcinomaでは髄様癌部にカルシトニンmRNA,蛋白共に発現しており,一方濾胞癌の部分ではサイログロブリンmRNA,蛋白共に発現していた.5.しかし,mixed medullary-follicular carcinomaの腫瘍細胞において,カルシトニン,サイログロブリンmRNA,蛋白は同一細胞において同時の発現は認められなかった.6.甲状腺髄様癌,甲状腺腺腫,乳頭癌等においても腫瘍細胞にカルシトニン,サイログロブリンmRNA,蛋白レベルとも同時発現は観察されなかった.<br />We studied to clarify the histogenesis of mixed medullary-follicular carcinoma of the thyroid using methods of clinicopathology, immunohistochemistry, in situ hybridization and northern blot. The results are as follows :1). Clinicopathologically, mixed medullary-follicular carcinomas are more frequently occurs compared with usual medullary carcinoma : in male patients (6 : 1 vs 1 : 1), in younger patients (36yrs vs 44yrs) and with better prognosis (100% vs 78% in 5-year survival).2). We failed to detection of calcitonin and thyroglobulin mRNA using northern blot, probably due to error of extraction of mRNA from thyroid tissues.3). Using in situ hybridization and immunohistochemistry, mRNA and protein of calcitonin or thyroglobulin are detected in mixed medullary-follicular carcinoma.4). We initially expected that mRNA or protein of calcitonin and thyroglobulin are concomitantly express in tumor cells of mixed medullary-follicular carcinoma. However, in the present study, such a concomitant expression of calcitonin and thyroglobulin in the same tumor cell was not observed.5). The concomitant expression of calcitonin and thyroglobulin was not observed in usual medullary carcinomas and other types of thyroid cancers.We will continue intensively the research for clarification of histogenesis of medullary-follicular carcinoma of thyroid.<br />研究課題/領域番号:10670156, 研究期間(年度):1998-1999 続きを見る
6.

論文

論文
水上, 勇治 ; Mizukami, Yuji
出版情報: 平成9(1997)年度 科学研究費補助金 基盤研究(C) 研究成果報告書 = 1997 Fiscal Year Final Research Report Summary.  1996-1997  pp.15p.-,  1998-03.  金沢大学保健学系
URL: http://hdl.handle.net/2297/00049262
概要: 今回の研究において私達は種々疾患の甲状腺組織を用いノーザンブロット法,in situ hybridization法および,モノクローナル抗体TSH受容体を用いた免疫組織化学的染色法を行いTSH受容体遺伝子のmRNAレベルと蛋白レベルにおける 発現状態を検索し,種々の甲状腺疾患における甲状腺濾胞細胞でのTSH受容体発現と濾胞細胞の機能さらに甲状腺ホルモン全体におよぼす影響を検討した.A.材料.1996年から1997年にかけて金沢大学第2外科にて手術で得られた甲状腺組織を対象とした.組織は切除後直ちに凍結して-80℃に保存された.B.プローブと抗体.TSH受容体遺伝子の検索に用いたプローブはTSH receptor cDNA vectorである。免疫組織化学染色に用いた抗体はマウス抗TSH受容抗体であり,これらはParis South大学Dr.E.Milgromより供与された.C.結果1.種々の疾患の人甲状腺組織を用い、TSH受容体遺伝子のmRNA発現、蛋白質発現をnothernblot法,in situ hybridization法,免疫組織化学法用い検討した.2.これら3種の方法を用いたTSH受容体発現は各疾患別にみてほぼ一致して結果が得られた.3.バセドー病甲状腺ではmRNAレベル,蛋白レベル共に発現が高度に増加し,バセドー病甲状腺におけるTSH受容体のup-regulation機構が作動していることが推測された.4.非機能性の甲状腺腺腫,乳頭癌においてもTSH受容体はmRNAレベル,蛋白レベルで発現が増加していた.5.未分化癌のTSH受容体はmRNAレベル,蛋白レベル共に発現が認められなかった.<br />In the present study, we investigated TSH-receptor expression in hyman thyroids using immunohistochemistry, in situ hybridization and northern blot. The relationship between the TSH-receptor expression and the thyroid function were examined.A.Materials and MethoedsThe surgically resected thyroid tissues of various thyroid diseases were used in this study. The thyroid specimens were immediately frozed at -80C and stored. The TSH-receptor cDNA vector and the anti-human mouse monoclomnal antibody against TSH-receptor protein were used. These were kindly supplied by Dr.E.Milgrom (paris South University, France).B.ResultsThe obtained results were as follows :1. The immunohistochemical, in situ hybridization and northern blot study showed the same results in the expression of TSH-receptor in human thyroids of each case. examined.2. In Graves' thyroids, the elevated TSH-receotor expression was observed in both of protein level and mRNA level. This finding supported the theory that the up-regulation mechanism of TSH-receptor is one of the major causes of Graves' hyperthyroidism.3. In the thyroid adenomas and the carcinomas which are functionally inactive, the elevated TSH-receptor expression was observed in the protein level and mRNA level.4. In the undifferentiated thyroid carcinomas, The TSH-receptor expression was not detected in the protein level and mRNA level.<br />研究課題/領域番号:08670196, 研究期間(年度):1996-1997 続きを見る
7.

論文

論文
山本, 美由紀 ; 井関, 尚一
出版情報: Acta Histochemica et Cytochemica.  36  pp.377-383,  2003-01-01.  日本組織細胞化学会
URL: http://hdl.handle.net/2297/6776
概要: 金沢大学大学院医学系研究科がん細胞学<br />The expression and localization of NGF and its high-affinity receptor, TrkA, in the rat ca rotid body was investigated by use of in situ hybridization and immunohistochemistry, respectively. The immunoreactivity for TrkA was detected in most of chief cells and sustentacular cells, as well as in some nerve axons and perineurial cells, but not in Schwann cells. In electron microscopy, the immunoreactivity was localized to the cytoplasm. The mRNA for NGF was localized to most of chief cells and possibly also to sustentacular cells. These results demonstrated that carotid body chief cells, having the neural crest origin, produce both NGF and TrkA, implying a paracrine-autocrine role of NGF in the carotid body. 続きを見る