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In vivo model mouse study to examine the effect of alteration of endogenous endocrine status in thyroid tumorigenesis

Research Project

Project/Area Number 18591030
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Endocrinology
Research InstitutionNagasaki University

Principal Investigator

NAMBA Hiroyuki  Nagasaki University, Graduate School of Biomedieal Sciences, Associate Professor (80237635)

Co-Investigator(Kenkyū-buntansha) NORISATO Mitsutake  Nagasaki University, Graduate School of Biomedical Sciences, Assistant Professor (50404215)
NAKASHINA Masahiro  Nagasaki University, Graduate School of Biornedical Sciences, Associate Professor (50284683)
Project Period (FY) 2006 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥3,890,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥390,000)
Fiscal Year 2007: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2006: ¥2,200,000 (Direct Cost: ¥2,200,000)
KeywordsThyroid cancer / Radiation / Oncogene / Animal Model / 動物モデル
Research Abstract

The aim of the study is to establish transgenic mice that develop thyroid cancer and then to examine the effects of a novel selective nuclear factor kappa B(NF-κB)inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ), which change various cytokines status in vivo, in the mouse model.
First Experiment : We prepared two types of transgenic mice. One is BRAF transgenic mice and another is Ret/PTC transgenic mice. Both transgenic mice develop papillary thyroid cancer within 4 weeks after birth. We examined the effect of DHMEQ, which is novel NF-κB inhibitor, to thyroid carcinomas developed in BRAF or RET-transgenic mice in vivo.
Results : Treatment with DHMEQ inhibited NF-κB DNA binding activity and induced apoptosis in dose and time dependent manner in thyroid carcinoma cells in vitro. However, in the in vivo study, DHMEQ could not suppress the NF-κB activity in thyroid carcinomas in the transgenic mice. Through the animal experiments, DHMEQ did not show any side effects in treated mice.
Second Experiment : We tried to establish novel thyroid carcinoma model mice using Cre-Lox-P system, which is allowed to express the interest gene at any time we want. We could establish the double transgenic mice that have Cre gene and BRAF-Lox-P gene. However, the mice could not develop thyroid carcinoma after induction.

Report

(3 results)
  • 2007 Annual Research Report   Final Research Report Summary
  • 2006 Annual Research Report
  • Research Products

    (61 results)

All 2007 2006

All Journal Article (33 results) (of which Peer Reviewed: 15 results) Presentation (27 results) Book (1 results)

  • [Journal Article] Different structural componentts of conventional papillary thyroid Carcinoma display mostly identical BRAF status2007

    • Author(s)
      Abrosimov A, et. al.
    • Journal Title

      International Journal of Cancer 120

      Pages: 196-200

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Characterization of side population in thyroid cancer cell lines:cancer stem-like cells are enriched partly but not exclusively2007

    • Author(s)
      Mitsutake N, et. al.
    • Journal Title

      Endocrinology 148

      Pages: 1797-1803

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] RET oncogene amplification in thyroid cancer:correlations with adiation-associated and high-grade malignancy2007

    • Author(s)
      Nakashima M, et. al.
    • Journal Title

      Human Pathology 38

      Pages: 621-628

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Clinical Implications of Pre-Operative Rapid BRAF Analysis for Panillarv Thyroid Cancer2007

    • Author(s)
      Kumagai A, et. al.
    • Journal Title

      Endocine Joumal 54

      Pages: 399-405

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Different structural components of conventional papillary thyroid carcinoma display mostly identical BRAF status2007

    • Author(s)
      Abrosimov, A., et. al.
    • Journal Title

      International Journal of Cancer 120・1

      Pages: 196-200

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Characterization of side population in thyroid cancer cell lines : cancer stem-like cells are enriched partly but not exclusively2007

    • Author(s)
      Mitsutake, N., et. al.
    • Journal Title

      Endocrinology 148・4

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] RET oncogene amplification in thyroid cancer : correlations with radiation-associated and high-grade malignancy2007

    • Author(s)
      Nakashima, M., et. al.
    • Journal Title

      Human Pathology 38・4

      Pages: 621-628

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Clinical Implications of Pre-Operative Rapid BRAF Analysis for Papillary Thyroid Cancer2007

    • Author(s)
      Kumagai, A., et. al.
    • Journal Title

      Endocrine Journal 54・3

      Pages: 399-405

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Different structural components of conventional papillary thyroid carcinoma display mostly identical BRAF status.2007

    • Author(s)
      Abrosimov A, et. al.
    • Journal Title

      International Journal of Cancer 120

      Pages: 196-200

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Characterization of side population in thyroid cancer cell lines: cancer stem-like cells are enriches partly but not exclusively.2007

    • Author(s)
      Mitsutake N, et. al.
    • Journal Title

      Endocrinology 148

      Pages: 1797-1803

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] RET oncogene amplification in thyroid cancer:correlations with radiation-associated and high-grade malignancy.2007

    • Author(s)
      Nakashima M, et. al.
    • Journal Title

      Human Pathology 38

      Pages: 621-628

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Clinical Implications of Pre-Operative Rapid BRAF Analysis for Papillary Thyroid Cancer.2007

    • Author(s)
      Kumagai A, et. al.
    • Journal Title

      Endocrine Journal 54

      Pages: 399-405

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] RET oncogene amplification in thyroid cancer : correlations with radiation-associated and high-grade malignancy.2007

    • Author(s)
      Nakashima M et al.
    • Journal Title

      Human Pathology (In press)

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Inhibition of ABL tyrosine kinase potentiates radiation-induced terminal growth arrest in anaplastic thyroid cancer cells2006

    • Author(s)
      Podtcheko A, et. al.
    • Journal Title

      Radiation Research 165

      Pages: 35-42

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Selective pharmacologic inhibition of c-Jun NH-2-terminal kinase radiosensitizes thyroid anaplastic cancer cell lines via induction of terminal erowth arrest2006

    • Author(s)
      Bulgin D, et. al.
    • Journal Title

      Thyroid 16

      Pages: 217-224

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] TP53 codon 72 polymorphism in radiation associated human papillarythyroid cancer2006

    • Author(s)
      Rogounovitch TI, et. al.
    • Journal Title

      Oncology Reports 15

      Pages: 946-956

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Childhood thyroid carcinoma with BRAFT1799A mutation shows unique pathological features of poor differentiation2006

    • Author(s)
      Kumagai A, et. al.
    • Journal Title

      Oncology Reports 16

      Pages: 123-126

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] No evidence of ARAF,CRAF,and MET mutations in BRAF(T1799A)negative human papillary thyroid carcinoma2006

    • Author(s)
      Kumagai A, et. al.
    • Journal Title

      Endocrine Journal 53

      Pages: 615-620

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] BRAFV600E promotes invasiveness of thyroid cancer cells through NF-kappaB activation2006

    • Author(s)
      Palona I, et. al.
    • Journal Title

      Endocrinology 147

      Pages: 5699-5707

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Pediatric CT scan usage in Japan:results of a hospital survey2006

    • Author(s)
      Ghotbi N, et. al.
    • Journal Title

      Radiation Medicine 24

      Pages: 560-567

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Inhibition of ABL Tyrosine Kinase Potentiates Radiatio-Induced Terminal Growth Arrest in Anaplastic Thyroid Cancer Cells2006

    • Author(s)
      Podtcheko, et. al.
    • Journal Title

      Radiation Research 165・1

      Pages: 35-42

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Selective pharmacologic inhibition of c-Jun NH2-terminal kinase radiosensitizes thyroid anaplastic cancer cell lines via induction of terminal growth arrest2006

    • Author(s)
      Bulgin, et. al.
    • Journal Title

      Thyroid 16・3

      Pages: 217-224

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] TP53 codon 72 polymorphism in radiation-associated Human papillary thyroid cancer2006

    • Author(s)
      Rogounovitch, T., et. al.
    • Journal Title

      Oncology Reports 15・4

      Pages: 949-956

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Childhood thyroid carcinoma with BRAFT1799A mutation shows unique pathological features of poor differentiation2006

    • Author(s)
      Kumagai, A., et. al.
    • Journal Title

      Oncology Reports 16・1

      Pages: 123-126

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] No Evidence of ARAF, CRAF and MET Mutations in BRAF(T1799A) Negative Human Papillary Thyroid Carcinoma2006

    • Author(s)
      Kumagai, A., et. al.
    • Journal Title

      Endocrine Journal 53・5

      Pages: 615-620

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] BRAFV600E Promotes Invasiveness of Thyroid Cancer Cells through NF-{kappa}B Activation2006

    • Author(s)
      Palona, I., et. al.
    • Journal Title

      Endocrinology 147・12

      Pages: 5699-5707

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Pediatric CT scan usage in Japan results of a hospital survey2006

    • Author(s)
      Ghotbi, N., et. al.
    • Journal Title

      Radiation Medicine 24・8

      Pages: 560-567

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Transworld Research Network2006

    • Author(s)
      H., Namba
    • Journal Title

      Recent Advance in Thyroid Cancer Research

      Pages: 61-91

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] TP53 codon 72 polymorphism in radiation-associated human papillary thyroid cancer.2006

    • Author(s)
      Rogounovitch T et al.
    • Journal Title

      Oncology Report 15・4

      Pages: 949-956

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Childhood thyroid carcinoma with BRAFT1799A mutation shows unique pathological features of poor differentiation.2006

    • Author(s)
      Kumagai A et al.
    • Journal Title

      Oncology Report 16・1

      Pages: 123-126

    • Related Report
      2006 Annual Research Report
  • [Journal Article] No Evidence of ARAF, CRAF and MET Mutations in BRAF(T1799A) Negative Human Papillary Thyroid Carcinoma2006

    • Author(s)
      Kumagai A et al.
    • Journal Title

      Endocrine Journal 53・5

      Pages: 615-620

    • Related Report
      2006 Annual Research Report
  • [Journal Article] BRAFV600E Promotes Invasiveness of Thyroid Cancer Cells through NF-{kappa}B Activation2006

    • Author(s)
      Palona I et al.
    • Journal Title

      Endocrinology 147・12

      Pages: 5699-5707

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Different structural components of conventional papillary thyroid carcinoma display mostly identical BRAF status.2006

    • Author(s)
      Abrosimov A et al.
    • Journal Title

      International Journal of Cancer 120・1

      Pages: 196-200

    • Related Report
      2006 Annual Research Report
  • [Presentation] Overexpression of wild-type THRB1 suppresses the growth and invasiveness of human papillary thyroid cancer cells2007

    • Author(s)
      Sedliarou Ilya
    • Organizer
      第80回日本内分泌学会学術総会
    • Place of Presentation
      東京
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] A case of thyroid hormone resistance with novel type of abnormal THRB1 transcripts2007

    • Author(s)
      Akulevich Natallia
    • Organizer
      第80回日本内分泌学会学術総会
    • Place of Presentation
      東京
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Peculiarities of distribution of genetic pattems of mutant BRAF in papillary thyroid carcinoma tissues2007

    • Author(s)
      Rogounovitch Tatiana
    • Organizer
      第80回内分泌学会学術総会
    • Place of Presentation
      東京
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Overexpression of wild-type THRB1 suppresses the growth and invasiveness of human papillary thyroid cancer cells2007

    • Author(s)
      Sedliarou, Ilya
    • Organizer
      The 80th JES Scientific Meeting
    • Place of Presentation
      Tokyo, Japan
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] A case of thyroid hormone resistance with novel type of abnormal THRB1 transcripts2007

    • Author(s)
      Akulevich, Natallia
    • Organizer
      The 80th JES Scientific Meeting
    • Place of Presentation
      Tokyo, Japan
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Peculiarities of distribution of genetic patterns of mutant BRAF in papillary thyroid carcinoma tissues2007

    • Author(s)
      Rogounovitch, Tatiana
    • Organizer
      The 80th JES Scientific Meeting
    • Place of Presentation
      Tokyo, Japan
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Overexpression of wild-type THRB1 suppresses the growth and invasiveness of human papillary thyroid cancer cells2007

    • Author(s)
      Sedliarou Ilya
    • Organizer
      第80回 日本内分努学会学術総会
    • Place of Presentation
      東京
    • Related Report
      2007 Annual Research Report
  • [Presentation] A case of thyroid hormone resistance with novel type of abnormal THRB1 transcripts2007

    • Author(s)
      Akulevich Natallia
    • Organizer
      第80回 日本内分泌学会学術総会
    • Place of Presentation
      東京
    • Related Report
      2007 Annual Research Report
  • [Presentation] Peculiarities of distribution of genetic patterns of mutant BRAF in papillary thyroid carcinoma tissues2007

    • Author(s)
      Rogounovitch Tatiana
    • Organizer
      第80回 日本内分泌学会学術総会
    • Place of Presentation
      東京
    • Related Report
      2007 Annual Research Report
  • [Presentation] Mutational and molecular epidemiology studies in radiation-associated human thyroid cancers2006

    • Author(s)
      Vladimir Saenko
    • Organizer
      日本放射線影響学会第49回大会
    • Place of Presentation
      札幌
    • Year and Date
      2006-09-08
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Mutational and molecular epidemiology studies in radiation-associated human thyroid cancers2006

    • Author(s)
      Vladimir, Saenko
    • Organizer
      The 49th Annual Meeting of The Japan Radiation Research Society
    • Place of Presentation
      Sapporo, Japan
    • Year and Date
      2006-09-08
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] SNPs profiling in post-Chernobyl thyroid cancers2006

    • Author(s)
      Namba H.
    • Organizer
      The 6th Meeting of the European Thyroid Association Cancer Research Network
    • Place of Presentation
      Naples,Italy
    • Year and Date
      2006-09-02
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] SNPs profiling in post-Chernobyl thyroid cancers2006

    • Author(s)
      Namba, H
    • Organizer
      The 6th Meeting of the European Thyroid Association Cancer Research Network
    • Place of Presentation
      Naples, Italy
    • Year and Date
      2006-09-02
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Re-expression of wild-type THRB1 in human papillary thyroid cancer cell lines suppresses growth and invasiveness of tumor cells2006

    • Author(s)
      Sedliarou I.
    • Organizer
      第79回日本内分泌学会学術総会
    • Place of Presentation
      兵庫
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Usefulness and significance of clinical application of pre-operative rapid BRAF analysis-results of Kazakhstan and Japanese clinical trial2006

    • Author(s)
      Kumagai A.
    • Organizer
      31st Annual Metting of the EUROPEAN THYROID ASSOCIATION
    • Place of Presentation
      Naples,Italy
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Therapeutic implication of miR17-5p and miR-19a for anaplastic thyroid cancer2006

    • Author(s)
      Takakura S.
    • Organizer
      31st Annual Meeting of the EUROPEAN THYROID ASSOCIATION
    • Place of Presentation
      Naples,Italy
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Radiation emergency medical project in Nagasaki University2006

    • Author(s)
      Akita S.
    • Organizer
      The 7th European Congress of Trauma and Emergency Surgery
    • Place of Presentation
      Malmo,Sweden
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Evaluation of Usage of Whole-body PET&PET/CT Cancer Screening in healthy Japanese People:Validity and Radiation Dose2006

    • Author(s)
      Iwanaga M.
    • Organizer
      The 1st World Congress on Ningen Dock
    • Place of Presentation
      Okinawa,Japan
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Singular BRAF status in structural components of PTC2006

    • Author(s)
      ウラジミール サエンコ
    • Organizer
      第49回日本甲状腺学会学術集会
    • Place of Presentation
      香川
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] BRAFV600E Promotes Invasiveness of Thyroid Cancer Cells through NF-kB Activation2006

    • Author(s)
      パローナ イリーナ
    • Organizer
      第49回日本甲状腺学会学術集会
    • Place of Presentation
      香川
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Re-expression of wild-type THRB1 in human papillary thyroid cancer cell lines suppresses growth and invasiveness of tumor cells2006

    • Author(s)
      Sedliarou, I
    • Organizer
      The 79th JES Scientific Meeting
    • Place of Presentation
      Hyogo, Japan
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Usefulness and significance of clinical application of pre-operative rapid BRAF analysis-results of Kazakhstani and Japanese clinical trial2006

    • Author(s)
      Kumagai, A
    • Organizer
      3lat Annual Meeting of the EUROPEAN THYROID ASSOCIATION
    • Place of Presentation
      Naples, Italy
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Therapeutic implication of miR17-5p and miR-19a for anaplastic thyroid cancer2006

    • Author(s)
      Takakura, S
    • Organizer
      31st Annual Meeting of the EUROPEAN THYROID ASSOCIATION
    • Place of Presentation
      Naples, Italy
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Radiation emergency medical project in Nagasaki University2006

    • Author(s)
      Akita, S
    • Organizer
      The 7th European Congress of Trauma and Emergency Surgery
    • Place of Presentation
      Malmo, Sweden
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Evaluation of Usage of Whole-body PET & PET/CT Cancer Screening in healthy Japanese People : Validity and Radiation Dose2006

    • Author(s)
      Iwanaga, M
    • Organizer
      The 1st World Congress on Ningen Dock
    • Place of Presentation
      Okinawa, Japan
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Singular BRAF status in structural components of PTC2006

    • Author(s)
      Vladimir, Saenko
    • Organizer
      The 49th Japan Thyroid Association
    • Place of Presentation
      Kagawa, Japan
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] BRAFV600E Promotes Invasiveness of Thyroid Cancer Cells through NF-κB Activation2006

    • Author(s)
      Palona, Iryna
    • Organizer
      The 49th Japan Thyroid Association
    • Place of Presentation
      Kagawa, Japan
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Book] Recent Advances in Thyroid Cancer Research2006

    • Author(s)
      H. Namba(分担著者)
    • Publisher
      Transworld Research Network
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary

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Published: 2006-04-01   Modified: 2016-04-21  

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