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AOKI Yasuhiro

    Graduate School of Medical Sciences Department of Forensic Medicine　Professor
Contact： aokiymed.nagoya-cu.ac.jp
Last Updated :2024/04/26

Researcher Information

URL

J-Global ID

Research Interests

  • 法病理学   法医人類学   法医遺伝学   

Research Areas

  • Life sciences / Forensic medicine

Education

  • 1984/04 - 1988/03  東北大学大学院  医学研究科  社会医学系
  •        - 1984  Gifu University  医学部  School of Medicine

Association Memberships

  • Japanese Society for DNA Polymorphism Research   Japanese Sociaty of Legal Medicine   

Published Papers

MISC

Research Grants & Projects

  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research
    Date (from‐to) : 2022/04 -2025/03 
    Author : 青木 康博; 福田 真未子; 琵琶坂 仁; 舟山 眞人
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research
    Date (from‐to) : 2019/04 -2022/03 
    Author : AOKI Yasuhiro
     
    We conducted a three-dimensional (3D) morphological analysis of the pelvis incorporating multivariate analysis and machine learning, including deep learning methods to develop new ex and age estimation strategies. We created 3D polygonal images consisting of the same number of points (facets) and the same topology and analyzed them using the methods mentioned above to verify the accuracy of sex and age estimation. As a result, a high accuracy rate of 99% to 92% was obtained for sex determination. On the other hand, for age estimation, the accuracy rate for the two group classifications was at most 75-80%, and the average error margin for age estimation by regression was ±9 years or more. Although the methods examined in this study could be feasible for forensic anthropological measurements, further research is required for the practical implementation.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research
    Date (from‐to) : 2016/04 -2019/03 
    Author : AOKI Yasuhiro; TANISJIRI Toyohisa
     
    Sexual dimorphisms and age-dependent morphological features of the human coxal bone were quantitatively analyzed using homologous models created from three-dimensional CT images of the pelvis (male: 514, female: 388 samples, age 16-100). Average coxal images of each sex and age decade were generated through principle component analyses (PCA). By measuring average point-topoint distances of 8472 corresponding points (ACPDs) between each homologous coxal image and the average images, the sex of more than 93% of the samples was correctly assigned. Some principal components (PCs) detected in PCA of the homologous models of the samples correlated fairly well with age and are affected by features of the curvature of the iliac crest, the arcuate line and the greater sciatic notch. However, neither multiple regression analysis using PCs related to age nor comparison of ACPDs with the average images of each age decade could produce accurate results for age decade assignment of unknown samples.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research
    Date (from‐to) : 2013/04 -2016/03 
    Author : Aoki Yasuhiro; FUKUTA MAMIKO
     
    Japanese population data of 323 female and 424 male samples on 27 loci of X-chromosomal short tandem repeats (X-STRs) were collected and analysed. A simulation study using the population data elucidated that X-STRs are useful to confirm the father-daughter relationships and female sibships sharing the father, namely full sibships and paternal half shibships. For the calculation of the likelihood ratio of female sibship, a linkage model on the basis of identity by descent approach were presented. In this model, 1) the linkage disequilibrium was taken into account regarding a cluster as a haplotype. and 2) the mutation was considered only when there was an apparent contradiction. This computation method is less complicated and could obtain reasonable likelihood ratios, therefore, should be effective to evaluate the kinship with X chromosomal polymorphic markers.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research
    Date (from‐to) : 2010 -2012 
    Author : AOKI Yasuhiro; BIWASAKA Hitoshi
     
    Three dimensional pubic bone images, synthesized with 3D scanner and reconstructed from multi-slice CT images, were analyzed to quantify any age-dependent morphological changes of the symphyseal faces. Average values of absolute mean curvatures of pubic symphyseal surfaces under 30 years old were higher than those of older individuals, of which the values were approximately constant. The ratio of the concave areas increased gradually with progressing age classes. In the prinsipal component analysis performed on the homology models of the pubic symphyseal faces, some components specific to each age class were detected. Furthermore, among the specimens of under 35 years old, a few age-correlated components were also identified.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research
    Date (from‐to) : 2006 -2007 
    Author : AOKI Yasuhiro; TAKAMIYA Masataka
     
    Dermal wound age provides forensic pathologists with valuable information for the reconstruction of crime scenes and determination of the cause of death. In this study, cytokine expressions in murine skins were analyzed using immunohistochemical methods, real-time quantitative PCR, and in situ hybridization techniques. Wound age estimation for human dermal wounds was also performed based on quantification of 17 cytokines. The potential parameters would be IL 5, IL 10, IL 12 p 70, IL 13, IL 17, GM-CSF, IFN γ, TNF α in the early phase of dermal wound healing, IL 6, MCP 1 in the middle phase, and IL 1 β, IL 2, IL 4, IL 8, G-CSF, and MIP 1β in the middle and late phases. In addition, an outsourced examination of IL 1 β, IL 4, IL 6, IL 8, IL 10, G-CSF, GM-CSF, IFN γ, MCP1, TNFα, which could be available to any forensic institute, was performed for confirmative purposes. Many factors have been proposed as markers for dermal wound age estimation, but the set of cytokines selected for the outsourced examination in the present study wound be useful in daily forensic practice.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research
    Date (from‐to) : 2005 -2006 
    Author : NAKAYASHIKI Nori; AOKI Yasuhiro; HASHIYADA Masaki
     
    1.In-the region 5'upstream-of the H19 gene, eight haplotypes composed of 14 SNPs were discovered to be applicable to the detection method of differentially methylated parental allele (DMPA). Application of DMPA detection method to new candidate SNPs in eight imprinted genes was failed. 2. Allele distributions of SNPs in four imprinted genes (H19, SNRPN, PEG3 and MEST) were examined. As a result, haplotype polymorphism of H19FR and SNPs in SNRPN and PEG3 revealed higher values of expected hetelozygosity. Multiplex PCR and mini sequencing method was used to simultaneous genotyping of above three loci plus HYMAI SNP. All seven SNPs in the four loci were successfully typed not only in native DNA but also in enzyme-digested DNA for discrimination of multilocus DMPA. 3. Methylated situation of CpGs around several SNPs in five imprinted genes was analyzed by using bisulfite-sequencing method at clonal level. In general, either paternal or maternal allele was expectedly classified into highly methylated or hypo-methylated situation according to each expression manner of the gene. However, some CpGs in the tested region showed opposite methylation status in several clones. Inconsistency of CpG methylation was considerably observed in PEG3 clones.
  • 日本学術振興会：科学研究費助成事業
    Date (from‐to) : 2005 -2005 
    Author : 金武 潤; 青木 康博; 舟山 眞人; 平岩 幸一; 那谷 雅之; 中園 一郎
     
    5県(岩手・宮城・福島・三重・長崎)の警察医会・検案医会に所属する医師(一部歯科医師を含む)を対象にプレゼンテーションを行った後、アンケート調査を行った。本システムに対する関心は高かったが、携帯電話のカメラ・ウェブ機能の利用率は低かった。 6大学(札幌医大・岩手医大・東北大・福島県立医大・三重大・長崎大)の卒業後3〜5年目の医師を対象にアンケート調査を行った。若手臨床医の多くが死体検案の経験とその指導を受けた機会が少なく、可能な限り死体検案を引き受けたくないという実態が判明した。費用に関しては、無料あるいは勤務先の病院が負担すべきとの意見が大半を占めたが、少数ながら個人負担でも構わないとする意見がみられた。その一方で、携帯電話利用に対する拒否反応はなく、本システムに対する需要と期待があることが確認できた 全国79大学の法医学教室を対象に、アンケート調査を行った。死体検案に関する電話相談件数は年間数件である施設が大多数を占めた。携帯電話による画像の質は助言を目的とする場合には十分であるが、受け手側の体制整備とセキュリティの確保が最大の問題であると指摘された。 セキュリティの確保については、パスワード、暗号化、指紋認証、遠隔情報削除機能、画像ファイルの自動消去といった既存の技術を組み合わせることにより可能と判断された。また情報の二次利用の制限についても考慮する必要性が認められた。 本システムの運用形態として、(1)病院単位、(2)個人利用、(3)警察署へ配備しての共同利用等が考えられるが、最も効果的なのは研修医の教育を目的とした病院単位での利用であると考えられた。本システムの基本構想は既に完成しており、運用ガイドラインの策定に着手し、実際の運用を想定したシステム開発および実証実験を行う段階にあると考えられた。
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research
    Date (from‐to) : 2002 -2003 
    Author : AOKI Yasuhiro; TAKAMIYA Masataka
     
    An animal experimental study was carried out to investigate availability of mRNA expressions of tissue-type plasminogen activator (tPA) and fibronectin, and inflammatory cell dynamics for wound age estimation of bruises. The bruises ware made by compressing the dorsal skin of mice with 2 iron plates with holes, 3mm in diameter. The mice were killed at 1, 3, 8, 24, 72, 144, or 240 hours post-injury. tPA mRNA expression peaked at 1 hour, and increased slightly at 72 hours post-injury. It was detected in epidermal cells, fibroblasts, and endothelial cells before and after injury, and from 3 hours in neutrophils and from 72 hours in macrophages, respectively. tPA was considered to play an essential role in the first step of tissue remodeling. A slight increase of tPA mRNA expression at later phase might be related to epidermal proliferation. On the other hand, expression of fibronectin mRNA was biphasic. It peaked at 1 hour, and significantly increase at 144 hours post-injury. Acute increase of fibronection would be due to tissue reaction of mechanical injury, and activation of coagulation cascade. Proriferation of the fibroblasts would be major cause of the late phase peak. Examination of the correlation of microscopic changes, such as inflammatory cell dynamics and epidermal thickening, with mRNA expression of tPA, fibronectin and other cytokines, would be informative for bruise healing and dating in the acute and chronic phases.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research
    Date (from‐to) : 2000 -2001 
    Author : AOKI Yasuhiro
     
    An animal experimental study was carried out to investigate availability of mRNA expressions of cytekines as markers for wound aging. A 2 cm full thickness incision was made on the dorsal skin of male DDY mice of 5 to 6 weeks old, and they were killed 0.5, 1, 3, 8, 24, 72, 144, or 240 hours after the incision. The expression of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) were examined immunohistochemically using specific antibodies. For wound age determination, bFGF, which was detected in the early 0.5- to 1-hour phases and the late 24- to 144-hour phases, seemed to be superior to VEGF because of its earlier expression and because of its more persistent expression in dermal fibroblasts with increasing postomortem interval. Therefore, expressions of bFGF mRNA during wound healing were subsequently analyzed using quantitative PCR and in situ hybridization techniques. Beside the dermal wound, mRNA dynamics of dermal, cerebral, hepatic and renal ones were also examined. bFGF mRNA expressions in the injured skin peaked at 1 hour, and they were detected in epidermal cells, fibroblasts, endothelial cells and neutrophils. Those of the injured cerebrum increased from 1 hour and peaked at 48 hours. The astrocytes were main source of vFGF mRNA supply in the injured brain. In hepatic and renal injuries, however, bFGF mRNA expressions increased slightly at 24 hours in endothelial cells, in neutrophils of the liver, and in glomeruli of the kidney. These results suggested that the mRNA dynamics of skin and brain derived from ectoderm were quite different from those of liver and kidney. In other words, time-dependent expressions of bFGF mRNA could be useful wound aging marker for the organs of ectodermal origin.
  • 日本学術振興会：科学研究費助成事業
    Date (from‐to) : 1999 -1999 
    Author : 中屋敷 徳; 熊谷 礼子; 青木 康博
     
    インプリンティング領域内における多型現象を法医学的に応用するべく検討した。染色体11p15.5におけるIGF2(父性アレル発現)ならびにH19(母性アレル発現)遺伝子内の塩基置換に基づく既知RFLP多型について,まず基本となるデータを得るために,岩手県集団の頻度調査および親子鑑定事例での遺伝様式を調べた。IGF2多型を報告にあるPCR-RFLP法に代えてPCR-SSCP法で調べたところ,簡単かつ確実な判定が可能となった(RFLP法による判定は時に誤判定を伴う)。一方,H19多型については,報告されているPCR-RFLP法が有効であった。それぞれの対立遺伝子頻度は報告されている数値と似ていたものの,期待値と観察値に若干の差が見られ,サンプル数を増やして追加・検討している。親子30例において遺伝的矛盾は全く見られなかった。 各々の多型の塩基配列に基づき、メチル化シトシン部位を推測してプライマーを合成した。DNAをbisulfite処理してmethylation specific PCR(MSP)を試みたが,両多型領域の特異的増幅は困難であった。これは,メチル化シトシン部位を特定するsequencingをすることで適切なプライマーデザインが可能となると考えられる。末消血中の両遺伝子の発現状態をmRNAのRT-PCRにより確認することと併せて,今後も研究を進める予定である。
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research
    Date (from‐to) : 1997 -1999 
    Author : AOKI Yasuhiro; NAKAYAMA Yumi
     
    To investigate mechanical effect of repeated minor impact to the pathobiology of the brain, 2 experimental head injury models were designed. In vivo, adult rats were injured utilizing weight-drop device consisting of brass weight(450g) falling through a acrylic guide tube(1m). After 3 insults at an interval of 24 hours, the brain were perfused with 4% Paraformaldehyde in phosphate buffer solution and removed. Histopathological and immunohistochemical studies disclosed neuronal injury in the cerebral cortex, and brain edema in the corpus callosum. Despite the lack of diffuse axonal changes, this model would be suitable for studying neuronal, changes associated with minor head trauma. In vitro model of axonal injury using cultured cells was designed to introduce traumatic alterations on neuronal processes and to identify mechanisms responsible for the formation of focal swellings by observation with phase-contrast and transmission electron microscopes. The culture dishes were oscillated one-dimensionally and horizontally 1, 2 or 3 times; repeating at intervals of an hour, 5 seconds a time; on a heat controlled shaker. The amplitude and frequency of the oscillation was 40 mm and 1 Hz, respectively. The injured processes showed two forms: the terminal increase in diameter of the processes and beading depending on injured portions. The microscopic finding suggests that the stress would destroy a cytoskeletal network, and then cause the spherical deformation of the processes. Ultramicroscopically, the beads contained compactly and sparsely misaligned cytoskeletons and prominently gathered densecore vesicles. The cytoskeletal reconstruction within the regenerated growth cones was also noted in the terminal swellings and beads. The destruction of these linkers would be responsible to initiate the cytoskeletal destruction. The initiating point of the cytoskeletal alterations, however, must be confirmed using three dimensional imaging of samples without soluble proteins and immunohistochemical study.
  • 日本学術振興会：科学研究費助成事業
    Date (from‐to) : 1995 -1995 
    Author : 青木 康博
     
    非放射性物質標識による核酸の検出を容易に行うことを目的として、チミン・ダイマーおよびペルオキシダーゼに特異的なbispecific抗体の開発を試みた。報告者は抗ペルオキシダーゼ抗体産生細胞株は確立・保持していたが、抗チミンダイマー抗体産生細胞株を保有していなかったため、単クローン抗チミン・ダイマー抗体を作製した。抗原にはサケ精子DNAに紫外線を照射してチミン・ダイマー化したものを用いBALB/Cマウスを免疫し、脾細胞をX63-Ag8-653と融合した。その結果数株のIgGおよびIgM抗体産生細胞を得たが、dot ELISA法でサケ精子DNAに対しても抗体活性を示すものが多く、特異性の高い株の再クローニング等を行っており、チミン・ダイマー化したブローブの検出に関しての検討は十分になしえなかった。一方合成DNA(ATTT)_5にUV照射したもの抗原としてin vitro免疫・細胞融合により抗チミン・ダイマー抗体作製を試みたが、抗体産生細胞株確立には至らなかった。これと並行して4量体によるbispecific抗体産生細胞株作製法について抗ペルオキシダーゼ抗体産生細胞株(P445)および抗A単クローン抗体産生細胞株(2A1)を用いて検討した。後者をウワバイン含有培地で馴化培養し、10^<-3>ウワバインに対する耐性株を選択し、さらに100μg/mlまでの6-アミノメルカプトプリン含有培地にて順次培養し、HAT感受性株を選択して前者と融合した。融合後一時的にbispecificityを有する細胞の発育が認められたが、経代培養中に抗体産生能を失うものが多かった。ハイブリドーマ同士の融合は、融合効率・増殖環境等に改良の余地があると考えられた。
  • 日本学術振興会：科学研究費助成事業
    Date (from‐to) : 1994 -1994 
    Author : 青木 康博
     
    ABO式血液型に対する単クローン抗体産生細胞株とペルオキシダーゼに対する抗体産生細胞株とを融合させ、血液型抗原および酵素と反応するbispecificな抗体の作製を試みた。報告者は単クローン抗酵素抗体産生細胞株を保有していなかったため、まず単クローン抗ペルオキシダーゼ抗体を作製した。抗原は市販の不活性化されたペルオキシダーゼを用いBALB/Cマウスを免疫した。その結果数株のIgGおよびIgM抗体産生細胞を得たが、このうち酵素と結合した際にその酵素活性を比較的阻害しない株(P445、IgG1)をハイブリドーマ融合に用いた。一方抗A・抗B単クローン抗体産生細胞株(2A1、B1)をウワバイン含有培地で馴化培養し、10^<-3>Mウワバインに対する耐性株を選択し、さらに100μg/mlまでの6-アミノメルカプトプリン含有培地にて順次培養し、HAT感受性株の選択を試みた。この操作によりB1についてはウワバイン耐性・HAT感受性株B1UHを確立することができたが、2A1については、培養中に抗体産生能を失うものが多く、現在のところ融合に好適なハイブリドーマを得ていない。 P445とB1UHを融合して4量体とし、ウワバイン・HAT加IMDM選択培地で培養を行い、2株のbispecific抗体産生細胞株PB9およびPB10を得たが、いずれの産生する抗体も抗B活性に比して抗ペルオキシダーゼ活性がやや低いため、現在再クローニング中である。この抗体は抗IgG・抗IgMのいずれとも反応する。酵素活性を阻害するか否かなどの詳細な検討は完了していない。2A1については現在もウワバイン・6-アミノメルカプトプリン含有培地で培養中である。一方ハイブリドーマ同士の融合は、4量体形成率の向上について技術的に改良の余地があると考えられた。さらに、今回扱った4量体細胞の増殖速度はハイブリドーマの1/2ないし1/3程度であり、抗体産生細胞株の確立に時間を要するのが問題点として挙げられる。 Monoclonal antibody ABO blood group Hybrid hybridoma
  • 日本学術振興会：科学研究費助成事業
    Date (from‐to) : 1993 -1993 
    Author : 青木 康博
     
    羊水中のタンパクの性状について検討し、羊水特異的にタンパク、特にABO式血液型物質担荷タンパクに対する単クローン抗体の作製を試みた。 臨床診断のため妊娠第16週の妊婦より得た羊水中のタンパクをSDS-PAGEにより分離し、免疫ブロッティングによりABO式血液型活性の有無について検討したところ、170KDaおよび200KDa以上の分画に活性を認めた。血清・唾液・精液・膣液等ではこれらの分子量領域には型活性がみられなかったので、この分画を羊水特異的なABO式血液型物質担荷タンパクを含む分画と考えた。O型羊水からゲル濾過によってこの分画を分離し、抗原としてBALB/c系マウスを免疫し、細胞融合・クローニングを行った。1回の免疫の抗原量は10μgで、週1回・3回腹膣内に投与し、細胞融合3日前にブ-スター投与を行った。アッセイはA型羊水を固相化したELISAによった。クローニングにより抗体生産細胞株を2株分離し、それぞれ産生された抗体の特異性について検討したところ、このうち1株が羊水と特異的に反応する抗体を産生していることが確認された。この抗体はIgG1抗体で全羊水と反応し、血清・精液その他の体液とは反応しないが、羊水中のどのタンパクに対し反応するか等の詳細な検討は完了していない。今後、抗体を精製し、その認識するエピトープについて検討するとともに、微量試料における羊水の証明および血液型検出等への法医学的応用を試みたい。

Social Contribution

  • 「女性なごや」に寄稿
    Date (from-to) : 2013/02-2013/02
    Sponser, Organizer, Publisher  : 行政
    ― 「女性なごや」に「名市大だより 入浴中の異状死について」を寄稿した。
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    Date (from-to) : 2012/05-2013/02
    Sponser, Organizer, Publisher  : 行政
    愛知県医師会館 愛知県警察、愛知県医師会とともに大規模災害時の検視・検案体制の整備について検討を行った。
  • 公開シンポジウムのシンポジスト
    Date (from-to) : 2012/06/09-2012/06/09
    Sponser, Organizer, Publisher  : 地域団体・NPO
    アクトシティ浜松 日本法医学会公開シンポジウム「東日本大震災における検案支援の実際と課題」のシンポジストを務めた。

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