RESEARCH ACTIVITIESGraduate School of Medicine
The University of Tokyo

March 2003

Assembled and Edited by
the Planning Committee for External Review
Hiroto Okayama
Nobutaka Hirokawa
Ryozo Nagai
Hirokazu Nagawa
Hiroshi Kurita
Susumu Wakai＜2＞Preface

The University of Tokyo Faculty of Medicine was established in 1877, under Japan's
modernizing Meiji government. Its roots go back to the vaccination center established with donations from 82 private practitioners in 1858, when Tokyo was still
called Edo, and over the past 145 years it has trained some 20,000 doctors.
While we are conscious and proud of our long tradition, we are acutely aware of theneed to reform our curriculum to meet the needs of a new age. We are determined toprepare our students to become tomorrow's leaders in international medicine
through the best possible undergraduate training.
So it is too in the realm of research, where we intend to uphold the highest standards
to solve the most challenging problems. For many years we have worked to consolidate and improve our strengths in this area.
The 1991 revision of Japan's Standards for University Establishment called on universities to examine and evaluate their own performance. Internal reviews have begun
at many universities. The newly established National Institution for Academic
Degrees began reviewing Japan's universities in 2000.
In undergraduate education, we now regularly carry out evaluations of our own programs, including through course evaluations by students. Formerly in research, weevaluated our work by publishing biannual lists of each department's publications;
we are taking steps to ensure that our evaluation will be more objective and rigorousin the future.
The designation in 1997 of the University of Tokyo as an institution centered on its
graduate schools was a turning point. It is vital that we give the future developmentof postgraduate education here a sound foundation. Universities must see themselves as they are seen by others. Hence the present external evaluation of ourresearch programs.
Research, teaching, and clinical practice are of course closely linked in a graduate
school of medicine, and all three require evaluation, yet this time, we restricted ourscope to research. Needless to say, this can under no circumstances be construed as
the relegation of teaching and clinical practice to ancillary roles. Simply, we hoped toreceive the most useful possible suggestions by narrowing our area of focus.
Research is of course, by itself, a vast field.

＜3＞

The review process was meticulously prepared and executed by a committee headedby Professor Hiroto Okayama. We invited world-renowned scientists of the highest
caliber to serve on the committee and review our faculty's work. Professor Erwin
Neher and nine other distinguished scientists from overseas worked in collaboration
with eleven eminent colleagues from Japan. I wish here to express my sincere gratitude to the evaluation team, who devoted so much of their valuable time to this task.
Whether internal or external, the principal meaning of any review lies in the willingness of those reviewed to listen and respond to fair criticism. Without vigorous
implementation of changes -- should any be shown to be necessary -- external review
will be sterile.
Greater openness is yet another major aim. Through outside evaluation we hope tomake our research activities as widely known as possible and to arouse the public's
interest in our work by publicizing both our findings and the outcome of peer review.
Exposure to outside scrutiny and a wider audience will encourage us to reflect on
future directions.
I earnestly hope that the findings of this external evaluation will, in these and manyother ways, be a powerful guiding force helping us to build better research structuresand programs at the University of Tokyo.
March 2003

Takaaki Kirino, MD, PhD
Dean, Graduate School andFaculty of Medicine
The University of Tokyo＜4＞

CONTENTS

Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .<3>
Planning and Implementation of the External Review . . . . . . . .<7>
Report of the External Review
Executive Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .<15>
Group Summaries
Group 1 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .<21>Group 2 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .<25>Group 3 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .<29>Group 4 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .<35>
Research Activities
Graduate School of Medicine, The University of Tokyo . . . . . . . . . .1
February 2003

＜5＞

＜6＞

Planning and Implementation of the External Review

In April 2002, on the decision of Dean Takaaki Kirino and with the agreement of seniorfaculty members, a planning committee for external review was formed. The planningcommittee comprised Professors Nobutaka Hirokawa, Ryozo Nagai, Hirokazu Nagawa,
Hiroshi Kurita, Susumu Wakai, and myself. We began designing the structure of theexternal review and selecting the reviewers. The purpose of the external review was toobtain information that would help us improve our research activities and maintainthem at the highest international level. All senior faculty members chairing departmentsin the Graduate School of Medicine were subject to review; they were asked to prepare
descriptions of the research activities of their group, providing a list of the current members of the department, an outline of the department's past and current research,
descriptions of up to five major research grants obtained in the past five years, and a listof no more than 50 of the best papers published by the group. These documents wereassembled and edited by the planning committee and printed as a single volume. In
addition, all reviewees were asked to provide copies of their ten most important papers.
The committee of reviewers comprised 21 experts of high international repute, 11 fromJapan and 10 from other countries, headed by Professor Erwin Neher. (The committee
members are listed on page 8.) The reviews covered activities in all areas of research,from basic and clinical medicine to public health and nursing. Each reviewee wasreviewed by the two most appropriate reviewers, one from Japan and one from anothercountry. In December 2002, each reviewer received a copy of the bound volume outlining research activities, copies of the ten most important papers of their reviewees, andinstructions regarding the procedures and scope of the evaluation. Each reviewer was
requested to begin an in-depth examination of their reviewees' research activities, on thebasis of these materials. On February 18 and 19, 2003, the external review committee visited the Graduate School of Medicine to hear presentations by all reviewees and to maketheir final evaluations. The presentations were grouped into four areas: basic medicine,internal medicine and related fields, surgical sciences, and social medicine and nursing.
Presentations in the four areas were given in parallel in the Medical Research andTeaching Building, as shown in the schedule on page 10.
All evaluations were collected at the end of the visit, and were sent immediately to
Professors Ito in Group 1, Saruta in Group 2, Kakita in Group 3, and Aoki in Group 4,
who were asked to prepare summaries for their groups, and to Professor Neher, who
prepared the executive summary. The summaries are published here.
Hiroto Okayama, MD, PhD
Chair
Planning Committee for External Review＜7＞

Alphabetical List of External Reviewers
Aoki, Yoshiki MD & PhD

Professor, Chairman and Director
Nagasaki University, Institute of Tropical Medicine, Japan"Parasitology"

Holzemer, William L. RN, PhD, FAAN Professor, Chair and Associate Dean for International
Programs
Department of Community Health Systems, School of
Nursing, University of California San Francisco, USA"AIDS Nursing Care"
Ito, Masao MD & PhD

Director
Brain Science Institute, RIKEN, Japan"Brain"Julien, Jean Pierre PhD

Professor
Department of Neurology, McGill University Faculty of
Medicine, Canada"Neuronal Intermediate Filaments and Amyotrophic
Lateral Sclerosis"

Kakita, Akira MD & PhD

Professor, Chairman and Hospital Director
Department of Surgery, Kitazato University School of
Medicine, Japan"Gastrointestinal Surgery and Liver Transplantation"

Kakizoe, Tadao MD & PhD

President
National Cancer Center, Japan"Urology and Bladder Cancer"

Kitamura, Yukihiko MD & PhD

Professor and Chairman
Department of Pathology, Osaka University Medical
School, Japan"Hematopoiesis, Hematopoietic Stem Cell and Tumor"

McCarley, Robert William MD

Professor and Head
Department of Psychiatry, Harvard Medical School, USA"Neurobiology and Behavior: Basic Sleep Research &Brain Imaging in Schizophrenia"

McGarvey, Stephen PhD

Professor of Community Health and Director ofInternational Health InstituteBrown University Medical School, USA"Human Population Biology and International Health"Neher, Erwin PhD

Professor and Director
The Membrane Biophysics Department, The Max-PlanckInstitut für biophysikalische Chemie, Germany"Patch Clamp and Exocytosis"

Nilsson, Kenneth PhD & MD

Professor
Department of Cellular Pathology, University of Uppsala,Sweden"Immunology, Tumor Biology and Virology"＜8＞

Nozawa, Shiro MD & PhD

Professor and Chairman
Department of Obstetrics and Gynecology, School of
Medicine, Keio University, Japan"Gynecologic Cancer"

Ogihara, Toshio MD & PhD

Professor, Chairman and Hospital Vice-Director
Department of Geriatric Medicine, Osaka University
Medical School, Japan"Hypertension"

Park, Jae-Gahb MD & PhD

Professor
Department of Surgery, Seoul National University Collegeof Medicine
President
National Cancer Center, Korea"Surgical Oncology"

Podolsky, Daniel K. MD

Mallinckrodt Professor of Medicine
Harvard Medical School, Massachusetts General Hospital,
USA"Hormonal Signaling in the Gastrointestinal Tract"

Saruta, Takao MD & PhD

Professor and Chairman
Department of Internal Medicine, School of Medicine,
Keio University, Japan"Hypertension, Adrenal Disorders and Water-Electrolyte
Balance"

Schmelzeisen, Rainer MD & DDS

Professor and Chairman
Oral and Maxillofacial Surgery, Plastic Surgery, UniversityClinic Freiburg
Albert-Ludwigs-University, Germany"Bone Metabolism and Cancer"

Sonenberg, Nahum PhD

Professor
Department of Biochemistry and McGill Cancer Center,
McGill University Faculty of Medicine, Canada"Eukaryotic Translation Control"

Toyoshima, Kumao MD & PhD

Director
SNP Research Center, RIKEN, Japan"Cancer Biology and Virology"

Tsumoto, Tadaharu MD & PhD

Professor and Chairman
Department of Neurophysiology, Osaka University,
Graduate School of Medicine, Japan"Neurophysiology and Optical Center"

Ueda, Reiko PhD

President
Okinawa Prefectural Collage of Nursing, Japan"Child Health Nursing"＜9＞

Schedule of External Review Presentation
Feb. 179:00-9:20

Introduction of reviewers (Tetsumon Memorial Hall)Group 1

Group 2

Room 1305
Room 1304B
Speakers
Reviewers
Speakers
Reviewers
9:30-10:00 Takao Shimizu,
Jean Pierre Julien Satoshi Kimura,
Daniel K. Podolsky
Cellular Signaling
Tadaharu Tsumoto Infection Control &Takao Saruta
Prevention
10:00-10:30 Hiroto Okayama,
Nahum Sonenberg Kuni Ohtomo,
Daniel K. Podolsky
Molecular Biology
Kumao Toyoshima Radiology
Takao Saruta

Group 3

Group 4

Room 1303
Room 1302
Reviewers
Speakers
Reviewers
Rainer Schmelzeisen Yasuki Kobayashi,
Stephen McGarvey
Shiro Nozawa
Public HealthYoshiki Aoki

Speakers
Yuji Taketani,
Obstetrics &Gynecology
Osamu Tsutsumi,
Gynecologic Surgery

10:30-11:00 Nobutaka Hirokawa,
Cell Biology & Anatomy

Jean Pierre Julien Nobumasa Kato,
Tadaharu Tsumoto Neuropsychiatry

Robert W. McCarley Kohei Hashizume,Toshio Ogihara
Pediatric Surgery

Rainer Schmelzeisen Kazuhiko Ohe,
Shiro Nozawa
Medical Informatics
and Economics
Tadashi Kakizoe
Hiroshi Kurita,
Akira Kakita
Mental Health

Coffee break (20min)
11:20-11:50 Hiroki Kurihara,
Physiological Chemistry

Jean Pierre Julien Shoji Tsuji,
Tadaharu Tsumoto Neurology

Robert W. McCarley Shinichi Takamoto,Toshio Ogihara
Cardiothoracic Surgery

Jae-Gahb Park
Tadao Kakizoe

Ichiro Kai,
Social Gerontology

11:50-12:20 Yasushi Miyashita,
Integrative Physiology& Metabolism

Erwin Neher
Masao Ito

Kazuhiko Yamamoto,Allergy andRheumatology

Daniel K. Podolsky Michio Kaminishi,
Takao Saruta
Gastrointestinal
Surgery

Jae-Gahb Park
Tadao Kakizoe

12:20-12:50 Kensaku Mori,
Cellular & Molecular
Physiology

Erwin Neher
Masao Ito

Masao Omata,Gastroenterology

Daniel K. Podolsky Masatoshi Makuuchi,
Takao Saruta
Hepato-BiliaryPancreatic Surgery

Jae-Gahb Park
Tadao Kakizoe

William L. Holzemer
Akira Akabayashi,
Health Economics/Health Reiko Ueda
Promotion Science/
Biomedical Ethics
William L. Holzemer
Yasuo Ohashi,
Biostatistics/Epidemiology Reiko Ueda
Epidemiology & Preventive
Health Sciences

Erwin Neher
Masao Ito

Toshiro Fujita,
Nephrology and
Endocrinology

Daniel K. Podolsky Tadaichi Kitamura,
Takao Saruta
Urology

Jae-Gahb Park
Akira Kakita

14:30-15:00 Masamitsu Iino,
Cellular & Molecular
Pharmacology

Erwin Neher
Masao Ito

Ryozo Nagai,
Cardiovascular
Medicine

Daniel K. Podolsky Hirokazu Nagawa,Takao Saruta
Surgical Oncology

15:00-15:30 Masayoshi Mishina,
Molecular Neurobiology

Erwin Neher
Masao Ito

Tomifusa Kuboki,
Psychosomatic
Medicine

Lunch (70min)
14:00-14:30 Tomoyuki Takahashi,
Neurophysiology

Stephen McGarvey
Yoshiki Aoki
William L. Holzemer
Reiko Ueda

William L. Holzemer
Reiko Ueda

William L. Holzemer
Reiko Ueda

Robert W. McCarley Kunihiko Tamaki,
Toshio Ogihara
Dermatology

Katsuya Kanda,
Advanced ClinicalNursing/ Nursing
Administration
Jae-Gahb Park
Keiko Kazuma,
Tadao Kakizoe
Adult Nursing/
Terminal & Long-term
Care Nursing
Rainer Schmelzeisen Chieko Sugishita,
Shiro Nozawa
Family Nursing

Kenneth Nilsson
Kazuhiko Nakahara,
Yukihiko Kitamura Clinical Laboratory
Medicine
Kenneth Nilsson
Yasuyoshi Ouchi,
Yukihiko Kitamura Geriatric Medicine/
Aging Research
Nahum Sonenberg
Kumao Toyoshima

Robert W. McCarley Kiyonori Harii,
Toshio Ogihara
Plastic & Reconstructive
Surgery
Robert W. McCarley Tsuyoshi Takato,
Toshio Ogihara
Oral & Maxillofacial
Surgery
Kozo Nakamura,
Orthopaedic Surgery

Rainer Schmelzeisen Sachiyo Murashima,
Shiro Nozawa
Community HealthNursing
Rainer Schmelzeisen Susumu Wakai,
Shiro Nozawa
International
Community Health
Jae-Gahb Park
Katsushi Tokunaga,
Akira Kakita
Human Genetics

William L. Holzemer
Reiko Ueda

Daniel K. Podolsky Makoto Araie,
Takao Saruta
Ophthalmology

Rainer Schmelzeisen Hiroshi Ushijima,
Shiro Nozawa
Developmental
Medical Sciences
Rainer Schmelzeisen Ryutaro Ohtsuka,
Shiro Nozawa
Human Ecology

Stephen McGarvey
Yoshiki Aoki

10:30-11:00 Takaaki Kirino,
Neurosurgery

Nahum Sonenberg Teruhiko Toyo-oka,
Kumao Toyoshima Organ Pathophysiology& Internal Medicine
Nahum Sonenberg Takashi Igarashi,
Kumao Toyoshima Pediatrics/Developmental Pediatrics
Jean Pierre Julien
Tadaharu Tsumoto

Jae-Gahb Park
Akira Kakita

Stephen McGarvey
Yoshiki Aoki

Coffee break (20min)
11:20-11:50 Joji Ando,
System Physiology

Kenneth Nilsson
Yukihiko Kitamura

Kazuo Hanaoka,
Anesthesiology

Tadao Kakizoe
Akira Kakita

Kenneth Nilsson
Yukihiko Kitamura

Naoki Yahagi,
Emergency & Critical
Care Medicine

Tadao Kakizoe
Akira Kakita

Coffee break (20min)
15:50-16:20 Masashi Fukayama,
Human Pathology
16:20-16:50 Kohei Miyazono,
Molecular Pathology
16:50-17:20 Kouji Matsushima,
Molecular Preventive
Medicine
Feb. 18
9:30-10:00

Akio Nomoto,
Microbiology

10:00-10:30 Tadatsugu Taniguchi,
Immunology

11:50-12:20 Shoogo Ueno,
Bioimaging &Biomagnetics
12:20-12:50 Kou Imachi,
Biosystem Construction& Control

Kenneth Nilsson
Yukihiko Kitamura

Lunch (70min)
14:00-14:30 Yasuo Ihara,
Neuropathology

Jean Pierre Julien
Tadaharu Tsumoto

14:30-15:00 Morihiro Sugishita,
Cognitive Neuroscience

Jean Pierre Julien
Tadaharu Tsumoto

15:00-15:30 Norio Suzuki,
Radiation Oncology

Nahum Sonenberg
Kumao Toyoshima

Coffee break (20min)
15:50-16:20 Ken-ichi Yoshida,
Forensic Medicine

Kenneth Nilsson
Yukihiko Kitamura

16:20-16:50 Kunio Shinohara,
Radiation ResearchInstitute

Nahum Sonenberg
Kumao Toyoshima

18:00-20:00 Reception at Seiyoken, 15th floor of University Hospital＜10＞

Robert W. McCarley Kimitaka Kaga,
Toshio Ogihara
Otorhinolaryngology,
Head and Neck Surgery
Fumio Eto,
Rehabilitation Medicine

Kiyoshi Kita,
Biomedical Chemistry

William L. Holzemer
Reiko Ueda
William L. Holzemer
Reiko Ueda

Stephen McGarvey
Yoshiki Aoki
Stephen McGarvey
Yoshiki Aoki

Stephen McGarvey
Yoshiki Aoki

Report of the External Review

＜11＞

＜12＞

Executive Summary

＜13＞

＜14＞

Executive Summary
March 2003

Prof. Dr. Erwin Neher
Chair
The External Review Committee

The University of Tokyo Graduate School and Faculty of Medicine is among the leading institutions of biomedical research and education in Japan. In anticipation of pending structural reforms the senior faculty members have decided to perform a review of
its research activities. For this purpose 21 experts from top biomedical institutions
throughout the world and Japan have been invited for a two-day evaluation session.
The conclusions and recommendations of this committee are being presented here. The
summary of results is subdivided into four groups, reflecting the review process,which was conducted by the four subgroups of the panel
Group 1 comprises 23 departments dedicated to the basic biomedical sciences, andresearch oriented clinical departments. The panel noted that the classical disciplines
of medicine have well been reorganized during recent years around the methodology and concepts of modern biomedical research. It was concluded that most departments are engaged in very original research, in which they are internationally highly visible, some of them leading their respective fields. Although the research work
reviewed was mainly basic in nature outstanding clinical implications wereobserved, which are expected to redirect clinical practice in the future. The panel
also identified a minority of research efforts for which reorientation and focusing
was recommended.
Group 2 consists of 13 departments in clinical medicine. These departments are quite
heterogeneous with respect to size and the subdivision of work between clinicaleducational and research duties. Research efforts include work on pathogenesis,
pathophysiology, diagnosis, therapy and prognosis of diseases in the respective
areas. The panel was impressed by the high academic motivation of the staff and bynumerous important publications in internationally highly reputed journals.
Furthermore, the panel was delighted to note fruitful collaborations with basic science departments within the University of Tokyo, but also with other facilities in＜15＞

Japan and with prestigious academic institutions abroad. Note was taken of abundant grant money from outside sources, reflecting superb academic achievements.
Regarding the overall organization of the clinical departments it was recognizedthat a better balance would be needed between the clinical and research staff withcareful consideration of workloads regarding patient care and educational tasks.
Such reorganisation should aim for adequate time for research activities in all areas.
On the other hand research activities in some departments were found to cover too
wide a range of topics. In such instances focusing and reorganisation is indicated.
Group 3 consists of 17 clinical departments centered around such fields as Surgery andReproductive, Developmental and Aging Sciences. The panel was impressed by thefact that each department has succeeded over the past decade to successfully
engage in basic as well as clinical research in spite of a ‘terribly busy clinical practice‘ at the University Hospital. Research activities of the vast majority of departments were judged as excellent with respect to originality, productivity, scientificimpact and clinical impact. In a few cases the panel pointed out that reorientation
towards more specific research topics with strong clinical implications might beindicated for clinical research units, in which not only research but also clinicalpractice is conducted on a daily basis.
Group 4 comprises 15 departments in the fields of Social Medicine, Health Science andNursing, and International Health. The panel concluded that, on the whole, theGraduate School and Faculty of Medicine at the University of Tokyo assumes aleading role in research in these fields in Japan. Some departments are regarded asworld leaders in their respective disciplines. It was pointed out that the Universityof Tokyo has pioneered in Japan in establishing Biostatistics and Biomedical Ethics
as newly required research fields in Health Research, and that professors of Nursing
have taken a national lead in many aspects of their discipline. However, it was also
recognized that the methods of the present review process may not be adequate to
fully appreciate the quality of some of the departments in Health Sciences andNursing. The panel noted that the International Health departments cover a wide
range of medical and epidemiological topics of relevance both to the developed anddeveloping world and concluded that the achievements apparent today clearly
identify the University of Tokyo as a promising center of excellence in research on
International Health. Closer inter-disciplinary and trans-disciplinary efforts willallow to further develop this role.
In summary it can be stated that the University of Tokyo Graduate School of Medicine
combines world-class research in the most important biomedical research fields withexcellent medical practice. It managed to make the transition in most disciplines from asolid-base traditional medical practice to research-oriented biomedicine, taking advantage of recent progress in molecular medicine and advanced biomedical engineering.
The University of Tokyo Graduate School of Medicine is committed to complete this＜16＞

process and is in a good position to do so, given the fact that it harbors world-classexperts in many of the relevant disciplines.
The panel felt compelled to emphasize that the strength of the Medical School lies in an
ideal balance between basic, knowledge-oriented research and applied or disease-oriented research - a balance, which emphasizes the importance of the former for the latter. Numerous examples were presented in which progress in basic research unexpectedly opened new insights into longstanding clinical problems, such as the discovery ofgenes responsible for malignant hyperthermia or for a hereditary neuropathy. It is in
the very nature of knowledge-generating research that problem-solving insights happen unexpectedly and that they occur the more often the more knowledge-oriented
research is allowed to follow its own path. The panel recommends that the Universityof Tokyo Graduate School of Medicine should continue to support its world-class biomedical research and this way build on its foundations for excellence in teaching andhealth care.
Addendum
The Process and Remit of the Review
Background: In view of pending structural reforms of the Japanese health care anduniversity system and considering dramatic advances in knowledge in the biomedical
sciences the Graduate school of Medicine of the University of Tokyo decided to conduct a review of its research efforts. The purpose of this exercise was supposed to be
twofold: First, to provide guidance for the faculty in pending decisions on the further
development of the School and second to document the international standing of thefaculty within the rapidly changing international research scene. The review was
intended to concentrate on research performance, with teaching and infrastructure
issues covered only to the extent, which was considered necessary for optimal research
conditions.
The Planning Committee: The review was prepared by a Planning Committee consisting of 6 senior faculty members and chaired by Prof. H. Okayama. The planning committee interacted with both the faculty and the review panel members by preparing thewritten materials (see below), formulating guidelines on the procedures of the review
process, and to guarantee an efficient review process within the time available.
Specifically, two panel members were assigned to each of the departments to bereviewed.
Materials and Documentation: Prior to the site visit panel members received twopieces of documentation besides general information regarding the University ofTokyo Graduate School of Medicine. One is a volume covering all of the departmentswith concise texts regarding their major research achievements, ongoing research, aswell as future plans. In addition it included a reference list of up to 50 select scientific＜17＞publications and a listing of personnel as well as current grant support. The other is acollection of the ten most important publications and evaluation sheets on thosedepartments, which had been assigned to a given panel member.
A schedule of the presentation and other events was mailed prior to the meeting. Onsite panel members received information material on the University as a whole and on
the Medical School. This included- The March 2002 Edition of ‘Tansei‘, The University of Tokyo Magazine- the 2003 prospectus of the Faculty of Medicine/Graduate School of Medicine- a site map
The review process: On the evening prior to the review, panel members met with the
Dean of the Medical School and the Planning Committee for a decision of the procedures and aims of the review. One of the panel members, E. Neher, was designated aschairman of the panel. Furthermore, four additional members (Profs. M. Ito, T. Saruta,
A. Kakita and Y. Aoki) were identified to cooperate with the chairman in the generation of the Review Summary Report by providing summary results of the four thematic subgroups (see page 19-37).
The main reviewing task was accomplished during the scientific presentations of the
68 departments, which were held in four parallel subgroups sessions on Monday,
February 17th and Tuesday, February 18th, 2003. Each head of department was allowed
20 minutes of presentation time followed by up to 10 minutes of discussion.
The report: The evaluation sheets (two from independent panel members for eachprincipal investigator) were made available to the chairman and to those responsible
for the summary reports of the four subgroups. The reports were sent after the panel
meeting to the chairman as the basis for this report. Drafts of the reports were circulated among the panel members and the final version was submitted to the Dean of theMedical School on 12th of March 2003.

＜18＞

Group Summaries

＜19＞

＜20＞

Group 1

Reviewers

Ito, Masao (RIKEN, Japan)
Julien, Jean Pierre (McGill University, Canada)
Kitamura, Yukihiko (Osaka University, Japan)
Neher, Erwin (Max-Planck-Institute, Germany)
Nilsson, Kenneth (Uppsala University, Sweden)
Sonenberg, Nahum (McGill University, Canada)
Toyoshima, Kumao (RIKEN, Japan)
Tsumoto, Tadaharu (Osaka University, Japan)

Summarized by

Ito, Masao

General Comments and Suggestions:
While reviewing the twenty-three departments of Group 1, we were deeply impressed by the highstandards and the extensive coverage of the topics presented to us. Through this review, we realize that theclassic disciplines of medicine such as anatomy, physiology, biochemistry, pharmacology and pathology
have been well reorganized during the past ten years around the methodology and conceptualization ofmodern biomedical research at molecular, cellular, system and cognitive levels.
We used a five-grade rating system for evaluation (A: excellent, B: very good, C: good, D: fair, E:poor) of each of the four aspects of research, namely, originality, productivity, scientific impact and general
aspect. Two assigned members of the review committee rated each of the 23 departments of group 1. The
highest grade, AA, was given to 13 departments for originality, 11 for productivity, 7 for scientific impact,
and 9 for general aspect. Hence, about one-half of the reviewed departments were deemed excellent. When
we counted all of the AA, AB and BB grades, 19 departments were evaluated as excellent or very good fororiginality, 19 for productivity, 17 for scientific impact and 20 for general aspect. We conclude that mostdepartments have chosen highly original research topics and conducted productive research with highscientific impact. These departments published numerous original papers in English, many of whichappeared in highly prestigious journals. The heads of the departments are well-established leaders and are
frequently invited to international symposia or lectures. We applaud the remarkable achievements of thereviewed departments and their heads, and wish that the Faculty of Medicine, University of Tokyo continues
to lead in biomedical research worldwide.
The research works we reviewed are mostly basic in nature. However, when the clinicalimplication was rated optionally, 16 departments were given at least one A grade. Therefore, we observe an
outstanding clinical implication in the research we reviewed, even though its strength varies fromdepartment to department.
We, however, note that six of the 23 reviewed departments received a low grade for one or more
aspects. Three of them receiving C grades may require improvement of their research performance. Two
departments received D grades, one for the relatively low productivity represented by the publication output
and the other for the lack of innovative approaches. Another department received E grades because its
research outcome was disappointing. We recommend that these three departments reconsider their researchplans and strategies.
Specific Comments and Suggestions for Research Activities in Each Field within the Group:
I. Cellular Signaling, Molecular Biology, Cell Biology and Anatomy, Physiological Chemistry &Metabolism, Integrative Physiology, Cellular & Molecular Physiology, Neurophysiology, Cellular &Molecular Pharmacology, and Molecular Neurobiology
Modern medicine is firmly based on biomedical research that clarifies molecular, cellular andsystem mechanisms of a normal living body. The nine departments in field I constitute a major core of suchresearch activities in the Faculty of Medicine, University of Tokyo. For the general aspect, four of them wererated with AA, another four with AB, and the remaining one with BB; hence, the nine departments of field I
were all evaluated as either excellent or very good.
Prof. Shimizu's group (Department of Cell Signaling) mainly focuses on an increasingly promisingarea, namely, lipid signaling. They cloned leukotriene A4 hydrolase, a key enzyme in the leukotriene
biosynthesis, and successfully produced various types of gene-manipulated mouse deficient in the1

leukotriene receptor, PAF receptor or cytosolic phospholipase A2, or mouse over-expressing the PAF
receptor. Prof. Okayama's group (Department of Molecular Biology) targets the cell cycle control,
differentiation control and anchorage-dependent growth, using the technology of the full-length cDNA
cloning, which Prof. Okayama and Prof. Paul Berg reported in 1982 and revolutionized the field of genecloning. This group continues to produce interesting results linking the cell cycle to oncogenesis. Prof.
Hirokawa (Department of Cell Biology and Anatomy) heads a large dynamic group whose productivity has
been truly outstanding. They have successfully analyzed motor proteins functioning in axonal and dendritictransport and identified most members of the kinesin gene family and clarified how they function as
molecular motors. They have made several epoch-making discoveries; e.g., a deficiency in KIF1 Bβ causes
type 2A Charcot-Marie-Tooth neuropathy and KIF3 knockout leads to the loss of the right-left asymmetry innormal body formation.
Using gene manipulation, Prof. Kurihara's group (Department of Physiological Chemistry &Metabolism) discovered that endothelin (ET-1) is required for the formation of craniofacial and cardiac
neural crest structures including brachial arches and great vessels, and that andrenomedullin (AM) is
indispensable for vascular morphogenesis during the development and regulation of blood pressure by NO
production. These provide the bases for their further work on the development of neural crest and vascularsystems. Prof. Miyashita's group (Department of Integrative Physiology) takes the challenge of clarifying
neural mechanisms of learning and memory, ingeniously combining molecular approaches with synaptic
plasticity, microelectrode recording from monkey brains and fMRI imaging of human brains. They
previously discovered specific memory neurons in the monkey ventral infratemporal cortex and more
recently have shown that these neurons operate using the brain-derived neurotrophic factor (BDNF). They
have recently demonstrated the top-down signals from the prefrontal cortex for retrieving memory from theinfratemporal cortex. Prof. Mori's group (Department of Cellular & Molecular Physiology) developed
genetic and imaging techniques for investigating specific neuronal connections that form the odor maps in
the olfactory system, including the olfactory bulb, pyriform cortex and olfactory tubercle. Their work is now
extended to clarify how specific connections are generated through neurogenesis by axonal recognition oftarget neurons.
Prof. Takahashi's group (Department of Neurophysiology) conducts excellent cell-physiologicalanalyses of synaptic transmission, using the unique preparation of the Calyx of Held, a synapse in theauditory pathway in which both presynaptic and postsynaptic compartments can be patch-clamped. This
laboratory masters such a technically demanding task hardly like any other laboratories, worldwide. Worksby Prof. Iino's group (Department of Cellular & Molecular Pharmacology) excels in its quantitativedescription of cellular processes related to IP3-induced calcium signals in neurons. They characterized
different subtypes of IP3 receptor playing different roles in various cell types. They have recently developed
genetically encoded sensors for IP3. Prof. Mishina' group (Department of Molecular Neurobiology) is
world-renowned for their contribution to the cloning of ion channel proteins, particularly in identifying
diverse NMDA receptor subtypes and their functional roles. His group identified the delta 2 type ofglutamate receptor, specifically located in cerebellar Purkinje cells. His group now attempts to combine
mouse and zebrafish genomics to obtain insights into brain development and function.
II. Pathology, Molecular Pathology, Neuropathology, Forensic Medicine, Molecular Preventive
Medicine, Microbiology, Immunology, and Neurosurgery
The eight departments constituting field II are dedicated to disease-oriented research that identifies
the causes of various diseases such as cancer, virus infections, heart or brain ischemia and Alzheimer's
disease, aiming at developing innovative medical treatments. For the general aspect, five of them were
evaluated as AA, another one AB, and the remaining two BB; hence, all of the eight departments of field II
are evaluated as excellent or very good.
Prof. Fukayama (Department of Human Pathology) conducts high-quality "niche" research on
Epstein-Barr virus-associated neoplasms and also on lung carcinoma. This area, however, needs to introduce
more new methodologies of molecular pathology. Their decision to merge with the Division of Surgical
Pathology of the University Hospital is reasonable and would allow more translational research. Prof.
Miyazono (Department of Molecular Pathology) developed his earlier world class work with Prof. C. -H.
Heldin to a new line of work on carcinogenic effects of the transforming growth factor-β (TGF- β). He is ina position to reformulate traditional pathology to molecular pathology, and appears to need more resources
and support. Prof. Ihara's group (Department of Neuropathology) focuses on molecular mechanisms
underlying Alzheimer's disease pathogenesis, and has made important contributions to the field ofamyloidogenesis with improvement of a method for quantifying amyloid β in human brains. Very recently,
2they have discovered the ε cleavage, distinct from the β- and γ-cleavage thus far established as themechanism underlying amyloidogenesis in the brain. Prof. Yoshida' group (Forensic Medicine) works onreactive oxygen generation, lipid peroxidation, and injury and death of the myocardium or brain after
ischemia repercussion or intake of toxic substances, which are subjects closely relevant to forensic medicine.
This group also aims to contribute to the legal aspects of forensic medicine, and to improve methods inforensic pathology. As an interface between the society and medicine, forensic medicine inevitably covers
broad areas, but for this group, a more focused approach may enhance its research activity.
The works on the molecular pathogenesis of inflammatory and immune diseases of Prof.
Matsushima's group (Department of Molecular Preventive Medicine) were deemed highly original, and if his
presentation had been better organized, the rate would have been even higher. Prof. Nomoto (Department ofMicrobiology), known for his pioneering work on poliovirus, is now extending his group's research toclarifying the mechanisms underlying the shut-off host protein synthesis by poliovirus. His work provides auseful model for hepatoma development in hepatitis C virus (HCV) patients, which is expected to be
applicable to other viral diseases as well. Prof. Taniguchi (Department of Immunology), renowned for his
pioneering work on cytokines, impressed the reviewers by the remarkable breadth and depth, andgroundbreaking significance of his group's research. It is hoped that the complex protein machinery in the
immune system that his group uncovered will be further analyzed using novel technologies (for instance,
cryoelectron microscopy). Prof. Kirino's group (Department of Neurosurgery) has made important
contributions to research on cerebral ischemia as well as on genetic analysis of tumors. They demonstrated
that the proteosome activity plays a key role in neuronal death after brief cerebral ischemia and that mice
deficient in NR2A NMDA receptors are less vulnerable to ischemic insult. A major breakthrough is thefinding that endogenous neuronal progenitors can be induced in situ to replace hippocampal neurons
damaged after ischemia.
III. System Physiology, Bioimaging & Biomagnetics, Biosystem Construction & Control, Cognitive
Neuroscience, Radiation Oncology, and Radiation Research Institute
These six departments are grouped together because of their two common features, namely, theyrequire large instruments and develop physical, rather than chemical, means for diagnosis and therapy. A
general difficulty in establishing this research field III, in spite of its importance as an interface between
biomedical research and clinical medicine, is reflected by the fact that the six departments received no A
grade for the general aspect. Three of them, nevertheless, received BB, that is, "very good", but theremaining three received low scores reflecting problems in their research plans or strategies.
Prof. Ando's group (Department of System Physiology) studies the effects of shear forces on
endothelial cells. The experimental device used to apply shear stress to cultured endothelial cells is valid,
and it provides the basis for their attempt to identify a flow sensor, shear-induced changes in gene
expression, and signal transduction in endothelial cells. The work, however, is still in a descriptive stage,
and a more focused approach may benefit them. The transcranial magnetic stimulation (TMS) performed by
Prof. Ueno's group (Department of Bioimaging and Biomagnetics) is in the front line of this field. Theirfinding that repetitive TMS facilitates regeneration or prevents damage to rat hippocampal neurons may
have important clinical implications. The biological effects of strong static magnetic fields on mammaliancells are interesting, and studies of these effects should be expanded using advanced technologies of cell
physiology, probably in collaboration with other departments. These basic studies may offer a basis forsecuring our health in environments where significant high-frequency electromagnetic waves emanate froman increasing number of commercial products such as mobile telephones . Prof. Imachi's group (Departmentof Biosystem Construction & Control) has inherited the long tradition of artificial heart engineering in the
Faculty of Medicine, University of Tokyo, and extends it to develop an implantable total artificial heart(TAH) that has so far enabled an implanted animal to survive for 63 days. This group will benefit fromcollaborations with clinicians as well as cell biologists, particularly if, as is suggested in their future plans,
they are going to start analyzing ES cells to generate whole organs. In view of the difficulty in hearttransplantation, in particular in Japan, a rapid progress of the TAH project has a great social significance.
Prof. Sugishita's group (Department of Cognitive Neuroscience) primarily focuses on the use of
MRI and magnetoencephalography (MEG) in studies of human cognitive functions. Mapping of brain
regions involved in writing and face recognition is the main contribution of this group. Since Prof. Sugishita
will retire very soon, reformulation of the aim of the Department including a new research plan is essential
for developing the important field of cognitive neuroscience. Prof. Suzuki's group (Department of Radiation
Oncology) group focuses on two research subjects, 1) radiation-induced signal transduction and apoptosis,
and 2) function of DNA protein kinases in double-strand- break repair and cell death. Their studies,3

however, appear to have diverged beyond the capacity of a relatively small number of researchers in thisgroup. It was also pointed out that not only the rather conventional approaches presently adopted but alsoinnovative methods should be introduced. Prof. Kunio Shinohara' group (Radiation Research Institute) is sosmall that the scope of its activity is limited. They developed a new technology of soft X-ray microscopythat may have some potential, but its usefulness has yet to be demonstrated.

4

Group 2

Reviewers

McCarley, Robert William (Harvard Medical School, USA)
Ogihara, Toshio (Osaka University, Japan)
Podolsky, Daniel K. (Harvard Medical School, USA)
Saruta, Takao (Keio University, Japan)

Summarized by

Saruta, Takao

General Comments and Suggestions:Group 2 consists of thirteen clinical medicine departments. They have different historicalbackgrounds, and the clinical and educational duties allotted to them also vary widely. Hence, their sizes andactivity scales are not uniform, and the number of the faculty members and the staffs differ considerably.
Regardless of the number of collaborators and the size of department, productive scientific research work withhigh originality has been conducted in each department within the framework of given conditions.
One of the important tasks of clinical departments is to conduct research on the pathogenesis,
pathophysiology, diagnosis, treatment, and prognosis of the diseases in each specialized area. The research
activities of the majority of the thirteen departments that underwent external review are being conducted withwell-defined objects and are considered satisfactory. It is well appreciated that in some of the thirteen
departments, a highly organized translational medicine-oriented approach is being taken to correlate theresearch and the clinical activities.
The majority of the staff of the departments that underwent review are highly motivated to act as theleading figures in this country through their clinical and research activities. The reviewers were impressed by
the high academic motivation of the staff reflected in the official compendium that introduces their research
activities. It is also impressive that numerous important publications in highly reputed journals have beenmade. In many instances, basic and clinical researches in these thirteen departments were performed incollaboration with basic science departments within the University of Tokyo, with other facilities in Japan, andalso with some prestigious academic institutions outside this country. Undoubtedly, such collaborative works
have been most rewarding. It should also be noted that these thirteen departments have received reasonably
abundant academic grant moneys likely reflecting their superb academic achievements
It is concluded that the research activities in the departments of group 2 are highly satisfactory ingeneral and that they should be encouraged to keep up with their current excellent performances. In some ofthe departments, however, the research activities have covered an excessively wide range of topics with rather
scanty organization. It is advised that the research activities in such departments should be concentrated on amore limited number of topics in the future.
In each of the departments in group 2, the staffs are expected to share various amounts of patientcare duties and education. It is felt that the number of the staffs in each department should better be rearranged
with careful consideration to the work loads that do not pertain directly to research activities: a larger numberof staff personnel should be secured for the departments with greater patient loads and educational tasks toallow an adequate time for research activities.
One final comment should be added to conclude this summary. This has been the first attempt toprovide a comprehensive external review of the research activities of the Graduate School of Medicine of the
University of Tokyo. Each of the professors was asked to make a presentation to the reviewers introducing the
research activities of his department. The form and the method of the presentation had not been predetermined
but were left to each of the presenters, resulting in presentations which differed considerably in organization,
length, and technical quality. It is possible that this situation exerted some influence on the final evaluations
made by the reviewers.
Specific Comments and Suggestions for Research Activities in Each Field within the Group:
1. DEPARTMENT OF INFECTION CONTROL AND PREVENTION:
This department has made important contributions in the prevention and control of hospital infections
within the university hospital as well as in other hospitals throughout Japan. It has also played an
important role as the center of diagnosis and treatment of HIV-infection in this country. The research
activities of the department have been focused on the prevention of MRSA infection, anti-HIV agents, andon basic mechanisms of hepatotrophic viral infections. In view of the significance of the clinical andscientific contributions being made by this department, it is recommended that the number of its faculties
1

be increased in the future.
2. DEPARTMENT OF RADIOLOGY:
The department of radiology has played a leading role in diagnostic radiology, radiation oncology, andnuclear medicine in this country through multiple clinical research projects. Applications of magnetic
resonance imaging and positron emission tomography in the diagnosis and studies on the pathophysiology
of various diseases have yielded contributions highly valued internationally. The department is expected tocontinue its current research and clinical activities, and is also anticipated to extend its efforts to somenewer fields of radiology including interventional radiology.
3. DEPARTMENT OF NEUROPSYCHIATRY:
Since 1994, when the ward and the ambulatory sectors of neuropsychiatry made the historical reunion, themembers of this department have been remarkably productive in their research activities. In particular,
their achievements in the study of post-traumatic disorders (PTSO) have been watched with keen interest.
Application of neuroimaging techniques to the study of human neuropsychiatric disorders is anotherimportant contribution made by them. Basic research works using animal models are also being pursued.
It is hoped that the department keeps up with its highly productive clinical and academic activities.
4. DEPARTMENT OF NEUROLOGY:
It is only six months since the current professor and chairman of this department took his office. The
department, however, is already making new important contributions in the study of molecularmechanisms of neurodegenerative disorders under the superb leadership of the new department chairman.
It is anticipated that the department will continue to play a leading role in the researches in molecular
genetics and gene therapy of neurodegenerative disorders.
5. DEPARTMENT OF CARDIOVASCULAR MEDICINE:
This department has been providing a comprehensive quality patient care in cardiovascular medicine.
Many basic research works have also been conducted, including a series of internationally valued studies
on the molecular biology of cardiac hypertrophy and cardiomyopathy. It is noteworthy that the membersof this department are attempting intensively to integrate the research and clinical activities.
6. DEPARTMENT OF GASTROENTEROLOGY:
This department has been actively involved in the study of pathogenesis, treatment, and prevention of viral
hepatitis and hepatocellular carcinoma that follows HCV infection, and it has undoubtedly played aninternationally recognized leadership in these fields. Important basic researches on the relationshipbetween Helicobacter pylori infection and gastric carcinoma have also been done. This department holds awell-organized system to undertake research works relevant to clinical gastroenterology. It is hoped that it
extends its effort further to the study of gastrointestinal tract malignancies besides gastric cancer.
7. DEPARTMENT OF NEPHROLOGY AND ENDOCRINOLOGY:
The research works of this department have covered a surprisingly wide range of topics yielding many
publications in the journals with high impact factors. However, it is the impression of the reviewer that the
research activities of the department have been spread over an excessively wide range of topics. The
department is advised to take a more organized approach in its research activities that enables strategic
handling of the selected subject matters by the collaborative works of its members.
8. DEPARTMENT OF ALLERGY AND RHEUMATOLOGY:
The research activities of this department have been translational medicine-oriented, and have dealt withthe pathogenesis of autoimmune disorders, basic immunology, and various clinical research works. The
publications made in the journals of basic immunology with high impact factor ratings are impressive,
although it is the opinion of the reviewer that more papers had better been submitted to the journals ofinternal medicine. The current research activity of the department, which is centered on the function of
antigen-specific T lymphocytes, is anticipated to yield discoveries highly important in immunology andimmunotherapy.

2

9. DEPARTMENT OF PSYCHOSOMATIC MEDICINE:
The members of this department have been actively involved in research on eating disorders and panic
disorders. Unfortunately, however, it is the opinion of the reviewer that their research efforts have notalways been most productive. Psychosomatic medicine should play an important role in our modern
society. It is encouraged that the members of this department continue with their efforts to makepublications in the journals with higher impact factor ratings and international reputations.
10. DEPARTMENT OF CLINICAL LABORATORY MEDICINE:
This department holds only two faculty members, both of whom serve concurrently as staff of the ClinicalLaboratory Center. This situation should be somewhat limiting to the research activities of thisdepartment. The research works of this department, however, have covered a surprisingly wide range of
topics. The reviewer is of the opinion that the research activities of this department should be concentrated
on a more limited number of topics. An increase of the personnel of this department should also be
seriously considered.
11. DEPARTMENT OF GERIATRIC MEDICNE/DEPARTMENT OF AGING RESEARCH:
This department was established in 1962 as the first geriatric medicine department of this country, and has
been held as an ideal model of the provider of medical care for the aged and for research in geriatric
medicine. The research works of the department have dealt with a wide range of topics including bonemetabolism, atherosclerosis and respiratory diseases. The opinion was expressed that the research
activities of this department had better been targeted on more selected subject matters relevant to geriatricpatient care.
12. DEPARTMENT OF ORGAN PATHOLOGY, INTERNAL MEDICINE AND HEALTH SERVICECENTER:
The members of this department are assigned to the health care of the faculty, the students and theemployees of the University of Tokyo. Therefore, the time they can spend for research should not be tooabundant. The research activities of this department have been focused on the molecular biology ofcardiac dysfunction. The department is expected to be involved in researches that have relevance tomass health care.
13. DEPARTMENT OF PEDIATRICS:
Pediatric medicine is expected to cover a wide range of health issues of children, and this department hasdone a remarkable job to bring this expectation to effect. Many important papers, both basic and clinical,
have been published in journals of high impact factor ratings. The department is expected to continue with
its currently superb research activities. It is also hoped that it extends its activity to the study of mental
health disorders of the children.

3

4Group 3

Reviewers

Kakita, Akira (Kitasato University, Japan)
Kakizoe, Tadao(National Cancer Center, Japan)
Nozawa, Shiro (Keio University, Japan)
Park, Jae-Gahb (National Cancer Center, Korea)
Shmelzeisen, Rainer (Albert-Ludwigs-University, Germany)
Kakita, Akira

Summarized byGeneral Comments and Suggestions;
The group 3 consisted of 17 departments in various clinical fields such as Surgical Sciences andpart of Reproductive Developmental and Aging Sciences of the University of Tokyo Graduate School of
Medicine. Five members of the review committee took part in the evaluation of these departments. Two of
the 5 members reviewed one department and each referee independently reported his reviewing result of theresearch activities of each department.
Through the entire review process, we were aware of and impressed by the fact that, under thecircumstances of terribly busy clinical practice in the University Hospital, each department had spared nopains to performing basic as well as clinical research during the past decade, despite many restrictions on thetime and money, the number of working staffs and research associates, and the installation for research
activities as well, in each department. In general, there were no remarkable differences among the reviewed
departments, although some departments had a long history as well as a bunch of research projects
accumulated since their foundation, whereas others had been instituted only recently.
Two assigned referees independently gave an assessment of the department after extensively
reviewing a self-documentation on the outline of past accomplishment, present research activities and future
prospect, as well as a hearing done for 2 days on research activities of each department. The referees used afive-graded scale for their evaluation (i.e., A: excellent, B: very good, C: good, D: fair, and E: poor) for eachof the following 5 categories: the originality, productivity, scientific impact, clinical impact, and general
rating. The highest mark AA was given to 9 departments for the originality, to 7 departments each for the
productivity, scientific impact and clinical impact, and to 10 departments for the general rating. All but one
department obtained marks of CC and superior for the 5 categories; only one department obtained the markCD for the category of clinical impact.
To inclusively assess departmentís research activity, scores from 5 to 1 were given to the gradesfrom A to E, respectively, and the sum of 2 refereeís scores in each category (e.g., AA=10, AB=9, BB=8,etc.) was classified into three ranks defined as excellent (the score 8 or larger), ordinary (the score 6 orlarger), and poor (the score less than 6). Thirteen departments were ranked as excellent and 4 departments asordinary for the category of originality. For the productivity, 15 departments were ranked as excellent and 2
as ordinary. For the scientific impact, 13 were evaluated as excellent and 4 as ordinary. For the clinicalimpact, 13 departments were ranked as excellent and 3 as ordinary, whereas 1 department was ranked aspoor. It may be noted that 15 of the 17 reviewed departments (88%) were ranked as excellent in the categoryof productivity, indicating that the prospects for further expansion of research activities in these departments
are promising.
Finally, looking at the assessment result for the category of general rating, 12 departments wereranked as excellent accounting for 70% of the 17 departments reviewed. Thus, it may be concluded thatmost department heads are well supervising their staff and fellow doctors including research associates, andare successfully directing their departments. For some of the remaining 5 departments ranked as ordinary,
however, it would take a bit more time to make any progress in their current research activities. In addition,
there was a suggestion from the reviewers to some of these departments that efforts might better be directed
toward the institution and execution of well-designed research projects on more specific subjects with strong
clinical impact, because these departments are regarded as clinical research units, in which not only basic
research but also clinical research and practice are conducted daily, and also because newly establisheddepartments very often have several limitations to their research environment.

1Specific comments and suggestions to each subspecialty group of Departments in group 3:
1. Surgery
Cardiothoracic Surgery; Gastrointestinal Surgery/Surgical Metabolism and Nutrition and Endocrine Surgery;
Hepato-Biliary-Pancreatic Surgery/Artificial Organs and Transplantation; Urology, and Surgical Oncology
For the category of general rating, all 5 departments in this group were ranked as excellent withthe average score of 8.8 (ranging from 7 to 10) in accordance with the scoring system defined above. It thus
appears that their research activities are to be favorably evaluated as excellent.
Department of Cardiothoracic Surgery (Takamotoís group) has made remarkable contributions to
the field of thoracic and cardiovascular surgery especially in aortic valvular and coronary artery bypass
surgeries, and thoracic surgery as well. In general, very good clinical setting-oriented research projects havebeen conducted to date. It is expected that many patients in the clinic would benefit from their continuous
research activities.
Dept. of Gastrointestinal Surgery (Prof. Professor Kaminishiís group) has focused its researchinterests on the pathophysiology and carcinogenesis of the gastrointestinal tracts system. The developmentof a model for creating chronic gastric ulcer, which was published in Gastroenterology, may be assessed as
an exceedingly unique and creative work. Also, a series of experiments related to Helicobacter pylori
infection and its eradication are very interesting. Research efforts in the future may better be exerted tovigorously continue the investigation on the carcinogenesis of gastric cancer.
Dept. of Hepato-Biliary-Pancreatic Srugery/Artificial Organs and Transplantation (Prof.
Makuuchiís group) has introduced and standardized the systematic subsegmentectomy of the liver, which is
emerging as the most significant idea in recent hepatic surgery, especially in cirrhotic livers. Prof. Makuuchi
also proposed the concept of preoperative portal vein embolization in the event of extensive hepatic
resection in patients with hilar bile duct cancer to minimize the postoperative hepatic dysfunction. Most ofall, he and co-workers are the first in the world to have performed adult-to-adult living related livertransplantation (LRLT), and have greatly contributed to improving the surgical techniques as well as thepost-transplant outcomes. Thus, their accomplishment may be favorably evaluated as excellent and theirendeavor to develop the hepatic surgery be respected, although they appear to attach too much importance toclinical issues.
Dept. of Urology (Prof. Kitamuraís group) has made good contributions to the understanding ofmultifocal carcinogenesis in the urinary bladder and also to the development of dendrite cell immunotherapy
for patients with metastatic cancer. Moreover, a series of studies on urinary polyoma virus JC virus for the
past decade have a great impact on the development of the diagnostic as well as therapeutic modalities.
These excellent findings, which had originated from well designed clinical as well as experimental studies,were recognized by several good international journals, and the results of this research is expected tocontribute to the advancement of the clinical urologic fields. There was a suggestion that these studies on the
carcinogenesis of urinary system should be continued and the knowledge obtained from this research should
be further directed toward clinical applications in the future.
Dept. of Surgical Oncology (Prof. Nagawaís group) has made a contribution to furthering not only
basic sciences such as immunology, molecular biology, and genomics, but also clinical oncology. It provides
comprehensive evaluation, diagnosis, treatment and management for adult patients with both general andoncologic surgical problems, in the ambulatory as well as inpatient settings. Among many excellent studiesof this department, the study that lateral nodes dissection is not necessary for patients with lower rectal
cancer undergoing preoperative radiotherapy is considered really important and very useful for treating
patients with advanced rectal cancer. These outstanding works would provide clinicians and practitioners
with valuable and novel ideas for managing patients. Prof. Nagawaís ardor for scientific knowledge andquality care of the patients should continue to be held.
2. Sensory and Motor System Medicine
Dermatology; Plastic and Reconstructive Surgery; Oral and Maxillofacial Surgery; Orthopedic Surgery;
Ophthalmology; Otorhinolaryngology, Head, and Neck Surgery; Rehabilitation Medicine
For the category of general rating, six of the 7 departments in this group gained the full mark, AA,or were ranked as excellent (the average score of 10 in accordance with the scoring system). The remaining
1 Department was assessed as ordinary, or as good with the mark, CC.
Department of Dermatology (Prof. Tamakiís group) appears as a well-structured, young

2

department showing good cooperation with other departments in the University, and also appears very
productive as indicated by its publications with a very high impact factor. This can be considered as a
successful transition from research to clinic. This group has shown a strong focus in the research front withincreasing applications of research results for clinical diagnosis. It has incorporated both cytokines researchand immunological research into the clinical realm. Research into the basic mechanisms of immunology isapplicable not only to Dermatology but to Cell Biology in general. Results of the research work on
metastasis of melanomas are similarly applicable to the mechanisms of metastasis in other malignancies.
Constructive criticism would be aimed at the application of the results of research works to the treatment ofdiseases in addition to the expanding molecular, enzymatic, and immunological diagnosis of dermatologicaldisorders. It is believed, however, that the direction of its perceptive studies would eventually incorporate
these goals towards actual clinical treatment of diseases.
Prof. Harii and co-workers (Dept. of Plastic and Reconstructive Surgery) have shown exceptional
ability to lead the vanguard in the still emerging field of reconstructive surgery. They have introduced new
surgical procedures and also have incorporated new techniques into the reconstruction of muscle and bonystructures for cancer patients. In addition, they have pursued new research leads into the molecular basis of
disease as well as the possibility of stem cell differentiation into tissues for skin and hair replacement. Their
outstanding research has had extremely strong impact on Plastic Surgery, and their excellent articles thatappeared in high-ranked journals have been appreciated worldwide. This clinical as well as basic researchshould be continued in cooperation with other departments. In particular, the possible use of epidermal andmesenchymal stem cells for replacement of various surface structures holds increasing promise for the
future. Optimization of their transformation and shaping/molding are topics for continuing research.
Moreover, their focus on the genetic basis of congenital anomalies related to plastic/reconstructive surgery iscommendatory. These supportive suggestions reconfirm the general positive trend of research carried out by
this Department.
Department of Oral and Maxillofacial Surgery (Prof. Takatoís group) has taken multi-pronged or
multidisciplinary approaches to both clinical and basic research studies that are very commendable. On theclinical front, in expanding the horizons of maxillofacial surgery, multidisciplinary approaches hold promisefor broadening treatment options for patients with deformities or tumors of the oral region. On the research
front, the collaborations with various academic as well as corporate groups are broad and varied, andcontinue to shape the future for reconstructive oral and maxillofacial surgery. The broad diversification ofresearch is to be lauded. This department has exhibited the delicate balance in stewardship necessary tofinesse the strengths obtained from both corporate and academic collaborations. This department has showna similar approach to translational research; the diversification of oral reconstructive surgery to regenerativetechnology and development are exemplary and show strong promise for the future.
Dept. of Orthopedic Surgery (Prof. Nakamuraís group) has done outstanding research in the fieldof orthopedics, especially of bone destruction, osteoclastogenesis in such areas as rheumatoid arthritis, senile
osteoporosis, and spinal cord or neuronal injuries. The departmentís research targets are very clear, andexcellent results of those research works were published in several prestigious international journals
including Nature. These results have been quite useful in the clinical orthopedic settings. This work is
believed to serve as a firm basis for deepening our understanding about orthopedic research. Studies on thepathogenesis of osteoclastogenesis and therapeutic tools for damaged spinal cord should be better continued
in the future.
Dept. of Ophthalmology (Prof. Araieís group) has shown a strong focus on both the clinical andbasic research forefronts of glaucoma pathophysiology and treatment. In clinical research, thepharmacokinetic evaluation and determination of pharmacological characteristics of various treatment
modalities for glaucoma are substantiated. In basic research, the investigations into the underlying molecularbiology and pathophysiology of glaucoma are similarly tangible foundations for future treatments. It may
appear that this department attaches too much importance to the evaluation of various treatment modalities
for glaucoma; an expansion of these developments and applications to other ophthalmologic disorders isstrongly suggested. Similarly, the research focus of the department is strongly directed toward glaucoma andrelated fields; collaborations with other fields of basic research would be a welcome addition to its research
portfolio. In this respect, the forays into corneal transplantation using regenerative medicine and amniotic
cell replacement are innovative and very promising.
Dept. of Otorhinolaryngology, Head and Neck Surgery (Prof. Kagaís group) has shown a strong
propensity for developing the products of research into innovative therapeutic modalities. On the clinicalforefront, this department continues to expand the pathophysiological understanding of the various sensory
organs while incorporating new treatment strategies towards malignancies. On the research forefront, the3

group continues to focus on the basic underlying mechanisms of disease that may lead to future clinicalapplications. Its work continues to be both broad in scope and deep. Its applications of research harvests toclinical treatments, especially in new substitutive modalities for the sensory organs, are intuitive andoriginal. Prof. Kaga seems to have taken strong leadership in publishing various articles as lead author
during his tenure; it is laudatory that he demonstrates the initiative in publishing to junior members of thedepartment.
Recognized as a new-borne department in clinical medicine field, the Department of
Rehabilitation Medicine (Prof. Etoís group) has been focusing its studies on the field of geriatric and stroke
rehabilitation. This group is especially interested in the functional and motor behavioral assessment ofgeriatric and brain-disordered populations, and has published several good relevant articles. In addition tocommitting themselves to clinical studies, Prof. Kaga and co-workers have recently broadened their research
spectrum to cover animal experiments, tissue and biomechanical studies. Because Prof. Eto has a strong
standing in the field of rehabilitation for geriatric people, research should better be continued in this field.
Moreover, it may be recommended that this department should institute a couple of more specific research
theme in this particular field of clinical medicine.
3. Vital Care Medicine
Anesthesiology; Emergency and Critical Care Medicine
The reviewers gave the mark BC each to these departments for the category of general rating,which is favorably evaluated as good or ranked as ordinary (the average score of 7 each) according to thescoring system.
Department of Anesthesiology (Prof. Hanaokaís group) has concentrated its energies mainly on the
topics and themes that were closely connected to the anesthesiology, namely, respiration and circulation,
pain management, muscle relaxation, development of new devices and techniques for anesthesia,
mechanisms of anesthesia, shock, and perioperative quality of life control. Above all, the establishment of
the guideline for postoperative pain management using local anesthetics combined with narcotics via
epidural catheter to the epidural space may be noted as a remarkable accomplishment of this department.
However, because of the multidisciplinary nature of research activities that this department is involved in,the objectives of basic as well as clinical research tend to be obscure and ambiguous. Therefore, it may berecommended that this department should institute much clearer objectives in the research field.
Dept. of Emergency and Critical Care Medicine (Prof. Yahagiís group) has only a small working
staff serving as clinicians and surgeons at both the Emergency Center and the Critical Care Center. In spiteof this awfully terrible condition, they not only have seen more than 17000 patients per year at theEmergency Ctr., but also have been engaged in many programs in a wide range of subjects related to basic
researches as well as practical problems. Particularly, their works on the structural parts involved in
activation and inactivation of Na channel that appeared in Nature (1989), and the clinical use of electrolyzed
water (Artificial Organs, 2000) are considered very commendable. Their curiosity to as well as enthusiasm
for research should not be discouraged from now on, and it is further suggested that they may better direct
their research interest toward more basic/scientific subjects.
4. Obstetrics and Gynecology
Obstetrics and Gynecology; Gynecologic Surgery
Both of these 2 departments gained the mark AA, or a full score of 10, for the category of general
rating. Thus, it appears that their research activities should be assessed and ranked as excellent.
In the Department of Obstetrics and Gynecology, Prof. Taketani has shown strong leadership in
supporting both diversification and specialization in his department. His research and clinical applications
for treatment of endometriosis are particularly noteworthy as well as his clinical prowess into the various
assisted reproductive technologies. To be noted is the fact that, in basic research field, this department has
been at the forefront of endocrinology and has expanded the knowledge of the underlying mechanisms offolliculogenesis and gametogenesis. Clinically, this department has recently shown strength in the field of
feto-maternal medicine; this is believed to be a strong focus for the department in the future. In addition, it isexpected that the fruits of these basic research works in endocrinology would find their way into clinicalpractice and technologies in the future.
Prof. Tsutsumi (Dept. of Gynecologic Surgery) has shown very active administration in both theclinical and basic research forefronts. This department has continued to lead the field in the application of4

endoscopic surgery for gynecologic disorders. Especially, research works of this department on thedetermination and surgical indications for lymph node spread in gynecologic malignancies are noteworthy.
In basic research, several works in molecular endocrinology and reproductive biology are very unique andintuitive. Any critical suggestions may not be necessary for this department. The work on environmental
estrogens and reproductive biology are perspective and influential; similar collaborations of Prof. Tsutsumiís
expertise in reproduction with experts in other fields may be recommended for the future.
5. Pediatric Sciences
Pediatric Surgery/Pediatric Oncology
Dept. of Pediatric Surgery (Prof. Hashizumeís group) has concentrated its research interests on
subjects in the following three areas, namely transplantation, developmental biology of congenital anomalies
and fetal surgery, and immunology of intestine. The idea of tracheal occlusion was very interesting, although
the clinical relevance was a little bit obscure. It may be recommended that the department should
concentrate its resources on experiments more related to clinical activities. In addition, considering the verysmall departmental staff, it may be suggested to select and institute a couple of much clearer research
objectives; experiments on beta-catenin, for example, seem to be too much.

5Group 4

Reviewers

Aoki, Yoshiki (Nagasaki University, Japan)
Holzemer, William L. (University of California, San Francisco, USA)
McGarvey, Stephen (Brown University, USA)Ueda, Reiko (Okinawa Prefectural Collage of Nursing, Japan)

Summarized by

Aoki, Yoshiki

General Comments and Suggestions:Group 4 includes three research areas encompassing fifteen different departments. The three researchareas are: 1) Social Medicine with two departments; 2) Health Sciences and Nursing with eight departments;and 3) International Health with five departments. Four reviewers were assigned to Group 4. Two reviewers
evaluated the academic achievements of seven departments of Social Medicine and International Health, while
the other pair evaluated eight departments within Health Sciences and Nursing. Evaluation followed themethods used by Groups of 1, 2 and 3. One reviewer assigned to Group 4 performed evaluations based solely
on documents prepared specifically for this external review by the University of Tokyo.
Inter-reviewer rating variation for the evaluation was relatively small among the group scores for all
departments. Of the fifteen departments of Group 4, twelve received the highest or second highest ratings, AA,
AB or BB. These results indicate that, on the whole, the Graduate School and Faculty of Medicine, Tokyo
University, assumes a leading role in research on Social Medicine, Health Sciences and Nursing, andInternational Health in Japan, while some departments are regarded world leaders.
A few departments within Health Sciences and Nursing received somewhat moderate or lower
ratings. Although these departments are encouraged to strengthen the quality and quantity of their research, it
may be that some, especially those belonging to Health Sciences or Nursing, have not been adequately
evaluated by the methods of the present external review.
Specific Comments and Suggestions for Research Activities in Each Field within the Group:
Subgroup I; Department of Public Health, Department of Medical Informatics and Economics
Subgroup I includes the Department of Public Health and the related Department of Medical
Informatics and Economics. Both departments have published often in well-regarded, peer-reviewed journals.
Accordingly, they are recognized as excellent or very good (AB, BB).
Public Health must embrace a wide range of interdisciplinary research. The research areas targetedby Department of Public Health are all essential topics, and are of special significance to Japan. Prof.
Kobayashiís group (Department of Public Health) presented research activities of high quality in varied,significant disciplines. This includes work on disabled elderly care system, tobacco control policy, the supplyof physicians and their demographic and geographic distribution, cardiovascular disease risk factors, sarin
poisoning, and other subjects. The long life expectancy and reduced birth rate has brought about a revolutionof health policy in Japan. The academic achievements of Department of Public Health indicate that Prof.
Kobayashi is making a significant contribution to an appropriate health policy for a changing Japan.
The department of Medical Informatics and Economics currently has large service component as
part of its overall mission in addition to its usual routine scholarly activities. Research interests within the
Department of Medical Informatics and Economics are basic to technical innovation in medical science andhealth care in this country. Prof. Oheís group (Department of Medical Informatics and Economics) has
initiated research on the development of a hospital informatics system and standardization of medical
information exchange. His group provides a useful information to medical staffs.
Subgroup II: Four departments relevant to Health Sciences and four departments relevant to Nursing
Sciences.
Health Sciences: Department of Mental Health, Department of Social Gerontology, Department of
Biomedical Ethics, Department of Biostatistics
Beyond traditional Health Science two new research fields, Biomedical Ethics and Biostatistics.
These have developed to reflect recent thinking about the social context of the conduct of research and newly
required research methods in health research. Anticipating that biomedical ethics and biostatistics will beessential to the evolving Japanese system of medical education across the many school of medicine in the near1

future, Tokyo University has assumed a leading role in these new areas of investigation. The decision toestablish these departments was a visionary one. Many useful papers have appeared in peer-reviewed journals.
However, scientific productivity of one department appears to be only moderate or low. Three departments are
given the mark of excellent or very good (AB, AB, BB) and one is rated as score of good (CC).
Prof. Kurita (Department of Mental Health) significantly contributes to the diagnosis and treatmentof pervasive developmental disorders. He is encouraged to continue promoting research reaching out tocollaborators in nursing and other fields of healthcare.
Professor Kaiís group focuses on health for the elderly, exploring fundamental human capacity,social support exchange, as well as the functional impact of home visiting and activities outside of the house.
Gerontology today is a multidisciplinary field and, although worthwhile, the scope of current efforts here is too
broad to be adequately addressed by the limited staffing now available. Professor Kai is encouraged to sharpen
his focus within the areas of functional capacity and quality of life for the aged.
Prof. Akabayashi (Department of Biomedical Ethics) is investigating the development of social
consensus, functions, and responsibilities of the ethical committee. He should play a vital role in developing anational code of biomedical ethics.
Prof. Ohashiís group (Department of Biostatistics) assumes a supportive role in clinical trials,
admirably serving the biostatistical needs of various principal investigators. Beyond this worthwhile activity
however, the department is challenged to become more engaged in their primary role, the development of new
statistical methodology for biomedical research, clinical trials, and epidemiology. They are also encouraged tolend their supportive expertise to research areas other than clinical trials.
Nursing Sciences: 1) Department of Advanced Clinical Nursing/Nursing Administration, 2)
Department of Adult Nursing/Terminal and Long-term Care Nursing, 3) Department of Family Nursing, 4)
Department of Community Health Nursing.
The professors of nursing have taken the national lead in nursing administration, in adult, terminal,
and long-term care nursing, and in family and community health nursing. There are some research areas which
do not appear to fit into the mission of the respective department. Many papers, published only in Japanese,would have a greater impact if incorporated into the English language literature. Two departments are given amark of excellent or very good (AB, BB), while two are rated as good (BC, CC).
Prof. Kandaís group (Department of Advanced Clinical Nursing/Nursing Administration) studies thenational nursing workforce, with particular emphasis on staffing ratios as predicators of healthcare quality.
They also address the measurement and improvement of quality of nursing care, This work has impacted
national healthcare policy in hospital staffing ratios.
Prof. Kazumaís group (Department of Adult Nursing/Terminal and Long-term Care Nursing) has
made an impressive use of the concentric circle model in research on muscle mass, exercise and fatigue. Her
focus on nursing care in an outpatient setting is an exciting development that is strongly encouraged.
Prof. Sugishitaís group (Department of Family Nursing) demonstrates national leadership in family
nursing. They have published useful investigations in such diverse areas as acupuncture, elderly quality of life,and physiological nursing care actions. However, little attention has been given to the family per se. The
department is encouraged to address both family and pediatric nursing.
Prof. Murashimaís group (Department of Community Health Nursing) offers useful insights into thecare delivery system, quality of care in the community health, and governmental policy. There appear to be
many opportunities for collaboration with the Department of Social Gerontology in the area of health for theelderly and with Nursing Administration in the areas of seamless hospital-community discharge.
Subgroup III: Department of International Community Health, Department of Human Genetics,
Department of Developmental Medical Sciences, Department of Human Ecology, Department of
Biomedical Chemistry
Subgroup III includes five departments whose interests focus on the field of International Health. It
covers a wide range of medical and epidemiological considerations, arising in both the developed anddeveloping world. International Health must embrace the continuing and widening economic and politicaldisparities that underlie poor health in developing nations. The research orientation of the chairman of eachdepartment is clear and appropriate and the research subjects targeted by each of the five departments are ofhigh priority in the field of International Health.
Even though Japan lacks a long history in research of International Health, the chairs of these fivedepartments, in their respective disciplines, have made considerable achievements in international health. All
departments have numerous publications in recognized journals. Two departments are recognized with thehighest scores, AA, and three are given a commendable AB.

2

Prof. Wakaiís group (Department of International Community Health) has made significant progressin the socio-economic, behavioral and biological basis of health using epidemiological and public healthanalytic techniques. Prof. Wakai is an effective advocate for the theoretical and applied aspects of International
Health.
Prof. Tokunagaís group (Department of Human Genetics) addresses some crucial areas of human
genetics, including anthropogenetic studies of associated populations, their histories, and the genetic basis of
disease. Their studies show that molecular genetics is a powerful tool with significant implications not only forInternational Health, but also more broadly for the future of biomedical science.
Prof. Ushijimaís group (Department of Developmental Medical Sciences) has done extensive workon molecular epidemiology of diarrheal diseases and on social epidemiology in maternal child health. Such a
program, linking basic science with maternal and child health, is an admirable innovation.
Prof. Ohtsukaís group (Department of Human Ecology) has made strides within a theoreticalframework linking environment, socioeconomic factors and human populations. Their studies arecharacterized by holistic and multi-disciplinary approaches.
Prof. Kitaís group (Department of Biomedical Chemistry) has demonstrated that basic research,
including molecular biochemistry represents a powerful set of tools holding great potential for public healthintervention. Their works on respiratory chains of eukaryote mitochondria and prokaryote cells may well leadto promising new treatments for parasitic diseases.
Achievements to date place clearly identify the University of Tokyo as a promising center of
excellence in the research of International Health. Nevertheless, close, inter-disciplinary and trans-disciplinary
collaborative efforts have not yet matured. We strongly recommend that these departments interact with one
other more effectively in order to develop their potential for world-class research in the field of International
Health.

3

＜38＞Graduate School of Medicine
The University of Tokyo

Assembled and Edited by
the Planning Committee for External Review
Hiroto Okayama
Nobutaka Hirokawa
Ryozo Nagai
Hirokazu Nagawa
Hiroshi Kurita
Susumu Wakai

Preface
The Graduate School and Faculty of Medicine of The University of Tokyo wasestablished in 1886 as the Imperial University Medical College. In keeping with
its tradition of leadership, it was recently modernized to increase the potential forfurther advances in research and education.
The Graduate School of Medicine has 66 departments, 4 closely associated facilities, and 6 endowed departments. The Graduate School grants doctoral degrees inbasic medical sciences, clinical medicine, and public health, and a master's degree
in medical science. A total of 200 graduate students are accepted each year. In
addition, the faculty members are fully responsible for teaching undergraduate
medical students. The departments are grouped into 11 programs: Molecular Cell
Biology; Functional Biology; Pathology, Immunology, and Microbiology;
Radiology and Biomedical Engineering; Neuroscience; Social Medicine; Internal
Medicine; Reproductive, Developmental, and Aging Science; Surgical Science;
Health Science and Nursing; and International Health. All are actively engaged inadvanced research and education.
With the goal of improving our research activities, the senior faculty members
recently decided to invite an external review. For this task, 21 internationallyrenowned scientists were selected to form the review committee.
This booklet outlines the past and current research status of each professor andeach research group. Committee members may find this background information
useful as they prepare to visit the campus and to hear the researchers' formal presentations. The visit and presentations will be held on February 17 and 18, 2003 atour Hongo campus. We greatly appreciate the committee members' willingness toshare their time and expertise, and we welcome their candid and constructive evaluations of our research activities.
February 2003

Takaaki Kirino, MD, PhD
Dean, Graduate School andFaculty of Medicine
The University of Tokyo

Alphabetical List of External Reviewers
Aoki, Yoshiki (Nagasaki University, Japan)
Holzemer, William L. (University of California, San Francisco, USA)
Ito, Masao (RIKEN, Japan)
Julien, Jean Pierre (McGill University, Canada)
Kakita, Akira (Kitazato University, Japan)
Kakizoe, Tadao (National Cancer Center, Japan)
Kitamura, Yukihiko (Osaka University, Japan)
McCarley, Robert William (Harvard Medical School, USA)
McGarvey, Stephen (Brown University, USA)
Neher, Erwin (Max-Planck-Institute, Germany)
Nilsson, Kenneth (Uppsala University, Sweden)
Nozawa, Shiro (Keio University, Japan)
Ogihara, Toshio (Osaka University, Japan)
Park, Jae-Gahb (National Cancer Center, Korea)
Podolsky, Daniel K. (Harvard Medical School, USA)
Saruta, Takao (Keio University, Japan)
Schmelzeisen, Rainer (Albert-Ludwigs-University, Germany)
Sonenberg, Nahum (McGill University, Canada)
Toyoshima, Kumao (RIKEN, Japan)
Tsumoto, Tadaharu (Osaka University, Japan)
Ueda, Reiko (Okinawa Prefectural Collage of Nursing, Japan)

4

CONTENTS
Molecular Cell Biology
1. Cell Biology and Anatomy
Department of Cell Biology and Anatomy .............................................................................................8
2. Biochemistry and Molecular Biology
Department of Molecular Biology ........................................................................................................14
Department of Cellular Signaling .........................................................................................................20
Department of Physiological Chemistry and Metabolism ...................................................................25

Functional Biology
1. Physology
Department of Integrative Physiology ..................................................................................................30
Department of Cellular and Molecular Physiology ..............................................................................35
Department of Neurophysiology ..........................................................................................................40
2. Pharmacology
Department of Cellular and Molecular Pharmacology ........................................................................45
Department of Molecular Neurobiology ..............................................................................................50

Pathology, Immunology and Microbiology
1. Pathology
Department of Human Pathology .........................................................................................................55
Department of Molecular Pathology ....................................................................................................60
2. Microbiology
Department of Microbiology .................................................................................................................65
Department of Infection Control and Prevention ................................................................................71
3. Immunology
Department of Immunology ..................................................................................................................76

Radiology and Biomedical Engeneering
1. Radiology
Department of Radiology ......................................................................................................................82
Department of Radiation Oncology (Experimental Radiology) ..........................................................88
2. Biomedical Engeneering
Department of System Physiology ........................................................................................................93
Department of Bioimaging and Biomagnetics .....................................................................................99
Department of Biosystem Construction & Control .............................................................................105

Neuroscience
1. Basic Neuroscience
Department of Neuropathology .........................................................................................................111
2. Speech and Congnitive Sciences
Department of Cognitive Neuroscience ..............................................................................................117
3. Clinical Neuroscience
Department of Neuropsychiatry ..........................................................................................................120
Department of Neurology ...................................................................................................................125
Department of Neurosurgery ..............................................................................................................132

5

Social Medicine
1. Occupational, Enviromental and Preventive Medicine
Department of Molecular Preventive Medicine .................................................................................138
Department of Public Health ...............................................................................................................143
2. Forensic Medicine, Medicine Informatics and Economics
Department of Forensic Medicine .......................................................................................................148
Department of Medical Informatics and Economics ..........................................................................153

Internal Medicine
1. Medicine I
Department of Cardiovascular Medicine ............................................................................................157
Department of Gastroenterology .......................................................................................................164
Department of Nephrology and Endocrinology ................................................................................170
2. Medicine II
Department of Allergy and Rheumatology ........................................................................................176
Department of Psychosomatic Medicine ............................................................................................182
3. Clinical Laboratory Medicine and Pathology
Department of Clinical Laboratory Medicine .....................................................................................188
Department of Transfusion Medicine .................................................................................................195

Reproductive, Developmental and Aging Sciences
1. Obstetrics and Gynecology
Department of Obstetrics and Gynecology ........................................................................................198
Department of Gynecologic Surgery ...................................................................................................202
2. Pediatric Sciences
Department of Pediatrics / Developmental Pediatrics .......................................................................205
Department of Pediatric Surgery / Department of Pediatric Oncology ............................................211
3. Aging Sciences
Department of Geriatric Medicine / Department of Aging Research ...............................................217

Surgical Sciences
1. Surgery
Department of Cardiothoracic Surgery ...............................................................................................223
Department of Gastrointestinal Surgery /Department of Surgical Metabolism and Nutrition and Endocrine Surgery ....................................228
Department of Hepato-Biliary-Pancreatic Surgery /Department of Artificial Organ and Transplantation ........................................................................234
Department of Urology .......................................................................................................................240
Department of Surgical Oncology .......................................................................................................245
2. Sensory and Motor System Medicine
Department of Dermatology ...............................................................................................................250
Department of Plastic and Reconstructive Surgery ............................................................................255
Department of Oral and Maxillofacial Surgery ..................................................................................258
Department of Orthopaedic Surgery ..................................................................................................264
Department of Ophthalmology ..........................................................................................................271
Department of Otorhinolaryngology, Head and Neck surgery .........................................................277
Department of Rehabilitation Medicine .............................................................................................282
3. Vital Care Medicine
Department of Anesthesiology ...........................................................................................................288
Department of Emergency and Critical Care Medicine .....................................................................293

6

Health Sciences and Nursing
1. Health Sciences
Department of Mental Health .............................................................................................................298
Department Of Social Gerontology ....................................................................................................303
Department of Health Economics / Health Promotion Sciences / Biomedical Ethics ........................307
Department of Biostatistics / Department of Epidemiology and Preventive Health Sciences .........311
2. Nursing Sciences
Department of Advanced Clinical Nursing / Department of Nursing Administration .....................316
Department of Adult Nursing/ Terminal and Long-term Care Nursing ............................................322
Department of Family Nursing ............................................................................................................327
Department of Community Health Nursing .......................................................................................332

International Health
1. International Social Medicine
Department of International Community Health ..............................................................................337
2. International Biomedical Sciences
Department of Human Genetics .........................................................................................................340
Department of Developmental Medical Sciences ..............................................................................344
Department of Human Ecology ...........................................................................................................350
Department of Biomedical Chemistry .................................................................................................356

Health Service Center
Department of Organ Pathophysiology and Internal Medicine Health Service Center ..................362

Radiation Research Institute......................................................................................................................368

7

Department of
Cell Biology and Anatomy
Outline and Research Objectives
When our institution was reformed from School of Medicine to Graduate School of
Medicine, Department of Cell Biology and Anatomy has been built with the transition fromthe previous department of Anatomy. Therefore, directions of researches in this department
are molecular cell biology based on structural cell biology.
At the same time this department has high responsibility for education of medical students, students of other faculties, and master course and Ph.D. course students (totally 518
hours per year). The teaching covers lectures and laboratory courses of gross anatomy, neuroanatomy, histology, cell biology and developmental biolgy.
Main subjects of researches are focused on 1) the mechanisms of intracellular transport,
2) the mechanism of cell morphogenesis, and 3) roles of cytoskeleton on development.
To solve these problems we combine structural biological approaches such as various
kinds of light microscopy, immunocytochemistry, quick freeze deep etch electron
microscopy, cryoelectron microscopy, X-ray crystallography with molecular biology, biochemistry, molecular biophysics, electrophysiology and molecular genetics.

Faculties and Students
Professor and Chair

Nobutaka Hirokawa, M. D., Ph.D.
(from 1983~)
Associate Professor Yoshimitsu Kanai, M. D., Ph. D.
Takao Nakata, M. D., Ph. D.
Lecturer
Yasuko Noda, M. D., Ph. D.
Sumio Terada, M. D., Ph. D.
Associate .................................8
Postdoctoral Fellow ................2 (Japan),1 (France),.................................................1 (China)
Graduate Students ..................7 (Japan), 2 (China)
Research Student ....................3 (China), 1 (Korea)
Secretary & Technician ...........4

Past Research, Current Research and Major
Accomplishments
I. Rapid Freeze Deep Etch Electron Microscopy
and Identification of New Groups of Crossbridge
Structures Among Microtubules, Intermediate
Filaments, Actin and Membranous Organelles.
Hirokawa has developed a new microscopic
method which rapidly freeze cells and tissues by making contact with a pure copper block cooled with liquid nitrogen (-190 C) or helium (-269 C) in Japan. He
further developed this method in USA with Dr. John
Heuser and visualised as yet unknown aspects of specialized membrane structures such as Gap Junction,
Tight Junction, and Neuromuscular Junctions. Further
he discovered a group of new filamentous crossbridges among microtubules, intermediate filaments,

8

actin filalments and membranous organelles.
Combining immunocytochemistry, biochemistry, andin vitro reconstitution he proved the chemical natureof these structures as 1) fodrin and myosin in brushborders of intestinal cells, a model of cellular cortex,
2) MAP1A, MAP1B, MAP2 and Tau in microtubule
domains in neuronal axon and dendrites, 3) elongated
C-terminus of Neurofilament M and H proteins inneurofilament domains in neuronal axons, 4)Synapsin I associated with actin and synaptic vesicles
in presynaptic terminals.
II. Discovery of Kinesin Superfamily Proteins,
KIFs and Elucidation of Mechanism ofIntracellular Transport.
Hirokawa discovered various kinds of new filamentous structures between distinct kinds of membranous organelles and microtubules. He predicted
these short crossbridges to be microtubule associatedmotor proteins carrying cargo vesicles along the
microtubules.
The nerve axon is frequently very long (frequently~1m long). Because of lack of protein synthesis
machinery in the axon most of the proteins necessary
in the axon and synaptic terminals ought to be transported down the axon as various kinds of membranous organelles and protein complexes after the synthesis in the cell body so that intracellular transport isfundamental for neuronal morphogenesis and functioning. At the same time because similar mechanism
exists in every kinds of cells nerve cells serve as agood model system to elucidate this mechanism.
Using molecular biology we discovered most ofkinesin superfamily motor proteins, KIFs which movealong microtubule rails by hydrolysing ATP. Recently
we identified all 45 kif genes in mammalian such as
human and mouse. We have uncovered the structure,dynamics and functions of many members of KIFs
usig molecular cell biological, biophysical, structural
biological and molecular genetical approaches.
KIFs in Axonal Transport
KIF1A and KIF1Bβ transport synaptic vesicle precursors anterogradely from cell biology to synaptic
terminals. They are fundamental for neuronal functioning and survivals. Further, we showed using
molecular genetic approaches that KIF1Bβ is a responsible gene of human hereditary neuropathy CharcotMarie-Tooth type 2A and proved that CMT2A is due
to haploinsufficiency of functional KIF1Bβ anddecrease of synaptic vesicle protein transport.
KIF1Bβ+/- mice serve as a model for CMT2A and
we found a way of diagnosis and potential therapy ofthe symptom of this desease. KIF1B a and KIF5A, B, C
redundantly convey mitochondria anterogradely.
KIF3, composed of KIF3A and KIF3B heterdimer andassociated protein KAP3, transports vesicles associated with a fodrin and important for neurites elongation through KAP3-α fodrin interaction. KIF2 andKIF4 are expressed specifically in juvenile neurons
and functioning for neurite extension.
Receptor Transport in Dendrites
Our studies revealed the mechanism of transportof receptors in dendrites.
KIF17 transports NMDA type glutamate receptor
containing vesicles in dendrites toward microtubule
plus ends through interaction between KIF17 C-terminal tail-Mint 1 (mLin10)- CASK (mLin2)- Velis (mLin7)
and NR2B subunit of NMDA receptor. Furthermore,
we showed by transgenic mouse strategy that KIF17
plays a significant role on memory and learning.
KIF5A, B, C convey AMPA type glutamate receptor
containing vesicles towards microtubule plus end
through the interaction between KIF5 heavy chain GRIP1 (glutamate receptor interacting protein 1)-GluR
2 subunit of AMPA type glutamate receptors.
These studies elucidated also for the first timelong standing questions how KIFs recognize and bindto cargoes by showing KIF tail - scaffolding protein
complex or adaptor protein complex- membrane protein interaction is a typical way. This idea was supported by our other study revealing that KIF13A
transports Mannose 6- phosphate Receptor containing
vesicles from Golgi to plasma mambrane through theinteraction between KIF13A tail - AP-1 adaptor protein complex and Mannose 6 phosphate receptors.
Our study about the transport of AMPA type receptor
by KIF5 also elucidated a mechanism how motor pro-tein determines its direction, axon vs dendrites.
GRIP1 steers KIF5 to dendrites via KIF5 heavy chain GRIP1 interaction, while JSAP1 steers KIF5 to axon
via KIF5 light chain- JSAP1 interaction.
KIF3 Determines Left-Right Asymmetery of OurBody
Our gene targeting study of KIF3 (KIF3A-/-, andKIF3B-/-) siginificantly contributued for the elucidation of mechanism of determination of left-rightasymmetry of our body, a very important hot problem
in developmental biology. Our study uncovered thatKIF3 motor is essential for left-right determination of
our body through intraciliary transportation of protein complexes for the ciliogenesis of motile primarycilia that generate leftward unidirectional flow of
extraembryonic fluid, "nodal flow", which could produce a concentration gradient of putative secreted
morphogen in the extraembryonic fluid along the leftright axis in the node of early embryo, very importantregion for the determination of our body plan.
Mechanism of Motility of Monomeric Motor,
KIF1A
We have discovered monomeric motors KIF1 subfamily such as KIF1A, KIF1B α and β. Because motors
previously identified are all dimers such as conventional kinesin (KIF5), dyneins and myosin the prevailing hypothesis for how the motor moves processively
for certain distance along rails was the hand overhand model, which means a motor needs two legs tomove as we walk on the rail. We, however, discovered
monomeric motor such as KIF1A. Because themonomeric motor is the simplest motor it is a goodmodel system to study the basic mechanism of motility of KIFs. We showed using molecular biophysics
approach that a single KIF1A motor can move processively for more than 1µm on microtubules.
Furthermore, our study revealed that the processive
movement of KIF1A is based on the biased Brownian
movement. This is the first clear experimental
demonstration showing that motor protein can moveby the biased Brownian movement. We further elucidated why single leg motor can move processively
along microtubules without dissociation. We foundpolylysin loop (K-loop) in loop12 and showed that
this K-loop is critical for processive movement
through the interaction with C-terminal flexible endof tubulin (E-hook) especially at the weak binding
state (ADP state) by biophysics, and cryoelectron
microscopy.
We further analysed how the plus end bias can be
emerged by combination of cryoelectron microscopy
and X-ray crystallography. We found that counterclock wise 20 degree rotation of motor domain from
ADP to ATP like state is critical for plus end biased

9

KIF review(Hirokawa 1998)

KIF17 transports NMDA
receptors in dendrites(Setou et al. 2000)

Kinesin structure(Hirokawa et al. 1989)

Photobleaching recovery
study of fluorescenttubulin (Okabe et al. 1990)

KIF1Atransports synaptic
vesicle precursor(Okada et al. 1995)

Tau induced microtubule
bundles and processelongation (Kanai et al. 1989)

KIF1B is responsible geneof hereditary neuropathy(Zhao et al., 2001)

Tau/MAP1B double
knockout mice
(Takei et al. 2000)

Cryo EM study of KIF1A motor
KIF1A monomer moves domain-microtubule complex(Kikkawa et al. Cell 2000)
processively by biased
Brownian movement(Okada &Hirokawa Science 1999)
KIFs transport various kinds ofcargoes in axon and dendrites

KIFs recognize and bind functional membraneproteins, through KIF
tail-scaffolding protein/adaptor protein complex interaction (Setou et al.
Science 2000, Nakagawa et al. Cell 2001, Setou et al. Nature 2002)

10

X-ray crystallography and cryoEM studyof monomeric KIF1A motor domain(Kikkawa et al. Nature 2001)

movement of KIF1A monomeric motor.
Mechanism of Slow Axonal Transport
Cytoskeletal proteins such as tubulin, actin andneurofilament triplet proteins (NFH, NFM, NFL) andsome cytosolic proteins were known to be transported down the axon slowly at 1~2mm/day. Using 1)
microinjection of fluorescein labelled cytoskeletal proteins and subsequent analysis by fluorescent photobleach recovery, 2) microinjection of caged-fluoresceinlabelled cytoskeletal proteins and subsequent analysis
by UV-photoactivation in cultured neurons weshowed most of cytoskeletal proteins such as tubulin
and actin are transported as small oligomers. Further,recently using microinjection of fluorescein labelledtubulin and cytosolic proteins into squid giant axons
and subsequent analysis with fluorescence confocal
laser scanning microscopy and fluorescence correlation spectroscopy combined with microinjection ofanti KIF antibodies we demonstrated that tubulin istransported as small oligomers by conventional
kinesin (KIF5) as a motor.
III. Microtubule Associated Proteins (MAPs) andMechanism of Neuronal Morphogenesis.
Hirokawa identified various kinds of filamentous
structures associated with microtubules by the quick
freeze deep etch electron microscopy. The next questions were what are the chemical nature and what are
the functions of these new structures. The microtubule associated proteins (MAPs) identified in brains
were good candidates. Therefore, we have studied
MAPs. We biochemically isolated major MAPs ofmammalian brains such as MAP1A, MAP1B, MAP2
and tau, and studied molecular structure and localization. All these MAPs were filamentous flexible strutures from 185 to 50nm in length dependent on the
difference of molecular weight. Immunocytochemistry
and in vitro reconstitution comfirmed that these
MAPs are components of filamentous structures associated with microtubules in neurons, To know thefunction of MAPs, we expressed tau and MAP2 in
non neuronal cells such as fibroblasts and Sf9 cells by
cDNA transfection. These studies showed that tau
and MAP2 induce microtubule polymerization andbundling and process extention. They also revealed
that C-terminal domain are critical for microtubule
polymerization and N-terminal projection domain
determines the different spacings between adjacentmicrotubules in microtubule domains in axon vs dendrites. These data suggest the important role of tau
and MAP2 in formation of axon and dendrites. In
order to elucidate function of MAPs in vivo we generated single knockout mice of tau, MAP1B, and MAP2
and analysed them. Generally, the phenotypes of single knockout mice were subtle. However, tau/MAP1B

and MAP2/MAP1B double knockout mice showed
predominant disturbance of axonal elongation anddendritic elongation respectively. Cellular bases ofthese phenotypes are suppression of microtubule stability and bundle formation in growth cones of axonand dendrites leading to the inhibition of proper neurite extention and neuronal cell migration. Thus, ourstudies revealed Tau/MAP1B and MAP2/MAP1B synergistically play fundamental roles in axonal and dendritic formation respectively.

Future Prospects
The further studies in near future will include following subjects as the continuation of on going
researches.
I. Mechanism of Intracellular Transport
1) The functions of new KIFs especially in neurons,
epithelial cells and fibroblasts to elucidate yet
unknown mechanism of intracellular transport ofvarious kinds of membranous organelles, protein
complexes and mRNA.
2) Understanding how each KIFs recognize and bindtheir specific cargoes.
3) The studies how the binding and unbinding of KIFs
with cargoes are regulated.
4) The mechanism how neurons determine the direction of transport by KIFs, toward axon vs dendrites.
5) The study of relationship between KIFs and neuronal functions such as memory and learning by
molecular genetic approach.
6) The relationship between KIFs and diseases using
molecular genetical approaches.
7) The detailed mechanism of motility of KIF1A motorusing structural biology and molecular biophysics.

II. Mechanism of Neuronal Morphophogenesis
and MAPs
Using molecular genetical approaches we will
unravel the function of MAP1A, one member of major
MAPs whose function has been unknown.

Research Grants
1. Center of Excellence (COE) grant from the Ministryof Education, Culture, Science and Sports, "Molecular
Cell Biological and Molecular Genetical Study of theCytoskeleton: Mechanism of Intracellular Transport,
Signal Transmission and Cellular Morphogenesis."
From April 1996 to March 2001. 1,885,000,000 yen
2. Grant in Aid for Basic Research, "Molecular Cell
Biology of New Molecular Motors, KIFs," from the
Ministry of Education, Culture, Science and Sports.
From April 1996 to Mach 1999. 34,500,000 yen
3. Center of Excellence (COE) grant from the Ministryof Education, Culture, Science, Sports andTechnology, "Mechanism of Intracellular Transport

11

:Molecular Cel Biology, Structure Biology andMolecular Genetics."From April 2001 to March 2006.
2,020,000,000 yen

Publication list
1. Hirokawa N and Kirino T. An ultrastructural study ofnerve and glia cells by freeze-substitution. Journal of
Neurocytology 9, 243-254, 1980.
2. Hirokawa N. Crosslinker system between neurofilaments, microtubules and membranous organelles infrog axons revealed by quick freeze, freeze fracture,
deep etching method. Journal of Cell Biology 94, 129142, 1982.
3. Hirokawa N, and Heuser J. E. The inside and outsideof gap junction membranes visualized by deep etching. Cell 30, 395-406, 1982.
4. Hirokawa N, Cheney R, and Willard M B. Location ofa protein of the fodrin-spectrin-TW 260/240 family
in the mouse intestinal brush border. Cell 32, 953965, 1983.
5. Hirokawa N, M. A. Glicksman, and M. B. Willard.
Organization of mammalian neurofilament polypeptides within the neuronal cytoskeleton. Journal of
Cell Biology 98, 1523-1536, 1984.
6. Shiomura Y and HirokawaN. The molecular structureof microtubule-associated protein 1A (MAP1A) invivo and in vitro. An immunoelectron microscopy
and quick-freeze, deep-etch study. Journal of
Neuroscience 7, 1461-1469, 1987.
7. Hirokawa N, Hisanaga S and Shiomura Y. MAP2 is acomponent of crossbridges between microtubules
and neurofilaments in vivo and in vitro. Quickfreeze, deep etch immunoelectron microscopy andreconstitutio studies. Journal of Neuroscience 8,2769-2779, 1988.
8. Hirokawa N, Pfister K K, Yorifuji H, Wagner MC,
Brady S T and Bloom GS. Submolecular domains ofbovine brain kinesin identified by electron
microscopy and monoclonal antibody decoration.
Cell 56, 867-878, 1989.
9. Kanai Y, Takemura R, Ohshima T, Mori H, Ihara Y,
Yanagisawa M, Masaki T and Hirokawa N.
Expression of multiple tau isoforms and microtubule
bundle formation in fibroblasts transfected with asingle tau cDNA. Journal of Cell Biology 109, 11731184, 1989.
10. Sato-Yoshitake R, Shiomura Y, Miyasaka H and
Hirokawa N. Microtubule-associated protein
1B:Molecular structure,localization, and its phosphorylation-dependent expression in developing neurons. Neuron 3, 229-238, 1989.
11. Okabe S and Hirokawa N. Turnover of fluorescently
labeled tubulin and actin in the axon. Nature 343,
479-482, 1990.
12. Hirokawa N, Yoshida T, Sato-Yoshitake R, andKawashima T. Brain dynein (MAP1C) localizes onboth anterogradely and retrogradely transported
membranous organelles. Journal of Cell Biology 111,
1027-1037, 1990.

12

13. Hirokawa N, Sato-Yoshitake R, Kobayashi N, Pfister
KK, Bloom GH, and Brady ST. Kinesin associateswith anterogradely transported membranousorganelles in vivo. Journal of Cell Biology 114, 295302, 1991.
14. Aizawa H, Sekine Y, Takemura R, Zhang Z, Nangaku
M and Hirokawa N. Kinesin family in murine central
nervous system. Journal of Cell Biology 119, 12871296, 1992.
15. Chen J, Kanai Y, Cowan NJ and Hirokawa N.
Projection domains of MAP2 and tau determine spacings between microtubules in dendrites and axons.
Nature 360, 674-677, 1992.
16. Hirokawa N. Microtubule organization and dynamics
dependent on microtubule-associated proteins.
Current Opinion in Cell Biology 6, 74-82, 1994.
17. Harada A, Oguchi K, Okabe S, Kuno J, Terada S,
Ohshima T, Sato-Yoshitake R, Takei Y, Noda T and
Hirokawa N. Altered microtubule organization insmall-calibre axons of mice lacking tau protein.
Nature 369, 488-491, 1994.
18. Kondo S, Sato-Yoshitake R, Noda Y, Aizawa H,
Nakata T, Matsuura Y and Hirokawa N. KIF3A is a
new microtubules-based anterograde motor in thenerve axon. Journal of Cell Biology 125, 1095-1107,
1994.
19. Sekine Y, Okada Y, Kondo S, Aizawa H, Takemura R
and Hirokawa N. A novel microtubule-based motor
protein (KIF4) for organelle transports, whose
expression is regulated developmentally. Journal of
Cell Biology 127, 187-202, 1994.
20. Nangaku M, Sato-Yoshitake R, Okada Y, Noda Y,
Takemura R, Yamazaki H, and Hirokawa N. KIF1B; A
novel microtubule plus end-directed monomericmotor protein for transport of mitochondria. Cell 79,
1209-1220, 1994.
21. Noda Y, Sato-Yoshitake R, Kondo S, Nangaku M and
Hirokawa N. KIF2 is a new microtubule-based
anterograde motor that transports membranousorganelles distinct from those carried by kinesin
heavy chain or KIF3A/B. Jounal of Cell Biology 129,
157-167, 1995.
22. Okada Y, Yamazaki H, Sekine Y, and Hirokawa N.
The neuron-specific kinesin superfamily protein
KIF1A is a unique monomeric motor for anterograde
axonal transport of synaptic vesicle precursors. Cell81, 769-780, 1995.
23. Kikkawa M, Ishikawa T, Wakabayashi T and
Hirokawa N. Three-dimensional structure of thekinesin head-microtubule complex. Nature 376, 274276. 1995.
24. Yamazaki H, Nakata T, Okada Y, and Hirokawa N.
KIF3A/B: A heterodimeric kinesin superfamily protein that works as a microtubule plus end-directed
motor for membrane organelle transport. Journal of
Cell Biology 130, 1387-1399, 1995.
25. Hirokawa N. Organelle transport along microtubulesthe role of KIFs (Kinesin superfamily proteins).
Trends in Cell Biology 6, 135-141, 1996.

26. Terada S, Nakata T, Peterson A and Hirokawa N.
Visualization of slow axonal transport in vivo.
Science 273, 784-788, 1996.
27. Saito N, Okada Y, Noda Y, Kinoshita Y, Kondo S andHirokawa N. KIFC2 is a novel neuron-specific C-terminal type kinesin superfamily motor for dendritictransport of multivesicular body-like organelles.
Neuron 18, 425-438, 1997.
28. Takei Y, Kondo S, Harada A, Inomata S, Noda T and
Hirokawa N. Delayed development of nervous system in mice homozygous for disrupted microtubuleassociated protein 1B (MAP1B) gene. Journal of Cell
Biology 137, 1615-1626, 1997.
29. Hirokawa N. Kinesin and dynein superfamily proteins and the mechanism of organelle transport.
Science 279, 519-526, 1998.
30. Nakata T, Terada S and Hirokawa N. Visualization of
the dynamics of synaptic vesicle and plasma membrane proteins in living axons. Journal of Cell Biology
140, 659-674, 1998.
31. Yonekawa Y, Harada A, Okada Y, Kanai Y, Takei Y,
Funakoshi T, Terada S, Noda T and Hirokawa N.
Defect in synaptic vesicle precursor transport andneuronal cell death in KIF1A motor protein-deficientmice. Journal of Cell Biology 141, 431-441, 1998.
32. Tanaka Y, Kanai Y, Okada Y, Nonaka S, Takeda S,
Harada A and Hirokawa N. Targeted disruption ofmouse conventional kinesin heavy chain, KIF5B,
results in abnormal perinuclear clustering of mitochondria. Cell 93, 1147-1158, 1998.
33. Nonaka S, Tanaka Y, Okada Y, Takeda S, Harada A,
Kanai Y, Kido M and Hirokawa N. Randamization ofleft-right asymmetry due to loss of nodal cilia generating leftward flow of extraembryonic fluid in micelacking KIF3B motor protein. Cell 95, 829-837, 1998.
34. Okada Y and Hirokawa N. A processive single-headed motor: kinesin superfamily protein, KIF1A.
Science 283, 1152-1157, 1999.
35. Takeda S, Yonekawa Y, Tanaka Y, Okada Y, Nonaka
S and Hirokawa N. Left-right asymmetry and kinesin
superfamily protein KIF3A: New insights in determination of laterality and mesoderm induction by
Kif3A-/- mice analyses. Journal of Cell Biology 145,
825-836, 1999.
36. Okada Y, Nonaka S, Tanaka Y, Saijoh Y, Hamada H
and Hirokawa N. Abnormal nodal flow precedes
situs inversus in iv and inv mice. Molecular Cell 4,459-468, 1999.
37. Okada Y and Hirokawa N. Mechanism of the singleheaded processivity : Diffusional anchoring between"k-loop" of kinesin and the C-terminus of tubulin.
Proceedings of the National Academy of Science of
the USA 97, 640-645, 2000.
38. Kikkawa M, Okada Y and Hirokawa N. 15Å resolution model of the monomeric kinesin motor: KIF1A.
Cell 100, 241-252, 2000.
39. Takeda S, Yamazaki H, Seog D-H, Kanai Y, Terada S
and Hirokawa N. KIF3 motor transports fodrin-associating vesicles important for neurite building.
Journal of Cell Biology 148, 1255-1266, 2000.

40. Setou M, Nakagawa T, Seog D-H and Hirokawa N.
Kinesin superfamily motor protein KIF17 and mLin10 in NMDA receptor-containing vesicle transport.
Science 288, 1796-1802, 2000.
41. Takei Y, Teng J, Harada A and Hirokawa N. Defects
in axonal elongation and neuronal migration in micewith disrupted tau and map1b genes. Journal of Cell
Biology 150, 989-1000, 2000.
42. Terada S, Kinjo M and Hirokawa N. Oligomeric tubulin in large transporting complex is transported via
kinesin in squid giant axons. Cell 103, 141-155,
2000.
43. Nakagawa T, Setou M, Seog D-H, Ogasawara K,
Dohmae N, Takio K and Hirokawa N. A novel
kinesin superfamily motor, KIF13A, transports mannose-6-phosphate receptor to plasma membrane
through direct interaction with AP-1 complex. Cell
10, 569-581, 2000.
44. Kikkawa M, Sablin EP, Okada Y, Yajima H, Fletterick
RJ and Hirokawa N. Switch-based mechanism ofkinesin motors. Nature 411, 439 - 445, 2001.
45. Zhao C, Takita J, Tanaka Y, Setou M, Nakagawa T,
Takeda S, Hong W-Y, Terada S, Nakata T, Takei Y,
Saito M, Tsuji S, Hayashi Y and Hirokawa N.
Charcot-Marie-Tooth Disease type 2A caused bymutation in a microtubule motor KIF1Bβ. Cell 105,587-597, 2001.
46. Miki H, Setou M, Kanashiro K and Hirokawa N. All
kinesin superfamily protein, KIF, genes in mouseand human. Proceedings of the National Academy of
Science of the USA 98, 7004-7011, 2001.
47. Teng J, Takei Y, Harada A, Nakata T, Chen J and
Hirokawa N. Synergistic effects of MAP2 and
MAP1B knockout in neuronal migration, dendritic
outgrowth, and microtubule organization. Journal of
Cell Biology 15, 65-76, 2001.
48. Noda Y, Okada Y, Saito N, Setou M, Xu Y, Zhang Z
and Hirokawa N. KIFC3, a microtubule minus enddirected motor for the apical transport of annexin
XIIIb-associated Triton-insoluble membranes.
Journal of Cell Biology 155, 77-88, 2001.
49. Setou M, Seog D-H, Tanaka Y, Kanai Y, Takei Y,
Kawagishi M and Hirokawa N. Glutamate-receptorinteracting protein GRIP1 directly steers kinesin to
dendrites. Nature 417, 83-87, 2002.
50. Harada A, Teng J, Takei Y, Oguchi K and Hirokawa
N MAP2 is required for dendrite elongation, PKA
anchoring in dendrites, and proper PKA signal transduction. Journal of Cell Biology 15, 541-549, 2002.

13

Department of
Molecular Biology
Outline and Research Objectives
This department was established in 1893 as the first Biochemistry Department in Japan.
Since then, this department has been devoted to biochemical studies on vitamin, oxidative
phosphorylation, lipid, carbohydrate and nucleic acid metabolisms, making remarkable contributions to advancements in medical biochemistry. After H. Okayama succeeded Chair in
1992, research has been focused on cancer, the cell cycle and differentiation in order tounderstand the molecular mechanism of malignant transformation and growth control.

Faculties and Students
Professor and Chair Hiroto Okayama, MD & PhD (1992~)
Associate .................................3
Postdoctoral Fellow ................1
Graduate Student....................5
Research Student ....................5
Secretary .................................2

Past Research and Major Accomplishments
Development of high-efficiency full-length cDNA
cloning method and eukaryotic expression cDNA
cloning system
Most of our past and current research stemmed
from the development of the full-length cDNA
cloning method and the cDNA expression cloning
vector system that was done in the early 1980s by
Hiroto Okayama and Paul Berg (Fig. 1)(1,2).
The full-length cDNA cloning method and thecDNA expression libraries constructed thereby havebeen used to clone numerous functional genes in laboratories over the world, the earliest one of which is
the human Lesch-Nyhan syndrome causative gene (3).
The cDNA expression cloning vector system was originally designed to clone genes on the basis of the

14

functions they express in a wide range of host cells,
from fission yeast up to mammalian cells. This wasenabled by development of highly efficient methods
for introducing cDNA libraries into mammalian cells
and fission yeast and construction of vectors specialized for library transduction into fission yeast (Fig. 1)(8, 10, 11). One of the most successful applications of
this expression cloning system was isolation ofhuman orthologues of several key cell cycle regulators
initially found in fission yeast. Paul Nurse cloned thehuman cdc2 cDNA from one of our expression
libraries on the basis of trans-complementation of afission yeast temperature-sensitive mutant of cdc2+
essential for mitosis, and we cloned human orthologues of wee1+ and cdc25+ genes (13, 14), which critically regulate Cdc2 kinase during this transition, as
described below.

Cell cycle control
Cdc2 kinase is a protein kinase required for the
onset of mitosis, initially found in fission yeast, but
present in all eukaryotes. This kinase, associated withcyclins, is regulated by inhibitory tyrosine phosphorylation and activating dephosphorylation by Wee1
kinase and Cdc25 phosphatase, constituting a coreelement of DNA damage-responsive cell cycle checkpoint control for the G2-M transition in fission yeast.

The identification of human orthologues (cdc2, wee1
and cdc25B) of these fission yeast cell cycle regulators
demonstrated the evolutionary conservation of a cell
cycle control system up to mammals (13, 14, 30).
Using the same expression cloning system, we also
identified an additional mammalian homologue(cdc25A) of cdc25+ gene, which was found to play an
unexpected role (23).
This gene is expressed in early G1 and required forthe cell cycle start. Search for targets for this novel
phosphatase led us to discover that Cdk4, a cyclindependent kinase essential for the cell cycle start, isregulated by tyrosine phosphorylation and that its
regulation constitutes a key part of DNA damageinduced G1 checkpoint arrest during cell cycle entry
from quiescence (23, 28, 37), revealing the presence of
mirror image mechanisms for controlling DNA damage-responsive G1 and G2 checkpoint arrest in mammals (Fig. 2).
In addition to the studies on mammalian cell cycle
control, we identified genes for 11 fission yeast novel
cell cycle factors and cyclin G a novel mammalian
cyclin (22), which control the G1-S transition, DNAreplication checkpoint or the formation and maintenance of sister chromatid cohesion (20, 24-27, 29, 33,
35, 40, 42-44, 48). Recent studies on two newly identified factors Eso1 and Pds5 involved in the formationof sister chromatid cohesion led us to discover thatsister chromatid cohesion is formed and maintainedby a highly unexpected mechanism, which functions
as a molecular zipper and thereby promotes the formation of cohesions only between sister chromatids(48). This finding provided a new clue to the understanding of the mechanism for the genetic instability
associated with malignant transformation.

Differentiation control
One of the aspects of the cell we have been interested in understanding concerning carcinogenesis isdifferentiation control. To this end, we also used fission yeast as a model organism and discovered fivenovel genes controlling the commitment to differenti-

ation. They encode Esc1 a MyoD-like helix-loop-helix
protein, Nrd1 an RNA-binding protein, Phh1 a stress
MAP kinase similar to p38, Rcd1 an evolutionally conserved protein and Pas1 and Cyc17 novel cyclins (21.
26, 31, 33, 36, 43, 44). These proteins were found to
be a modifier of cyclin AMP signaling, a determinantof the threshold to nutrient starvation for the commitment to differentiation, a stress signal transducer
as an absolute requirement for the differentiationcommitment, a key mediator of nitrogen starvation
signal and factors that ensure mutual exclusivenessbetween the cell cycle start and the commitment todifferentiation. In addition, using trans-complementation cloning, we isolated a human functional counterpart of the RNA-binding protein with predicted functions, which were shown in hematopoietic cells (38).
Furthermore, we recently found that the mammalian counterpart of Rcd1 is a novel transcriptional
cofactor mediating retinoic acid-induced cell differentiation and is deeply involved in mouse lung development (50). These findings not only reveal remarkableconservation of the differentiation commitment system as well as the key elements that control the commitment step from yeast up to mammals (Fig. 3), butalso greatly help understand the highly complex
mechanism controlling the differentiation commitment in mammals.

Anchorage-independent cell cycle start: a key
mechanism for malignant transformation
One of the fundamental phenotypes that distinguish malignant from benign cells is the metastatic
capability, which is underlain by the acquirement of
the ability to perform S phase entry in the absence ofanchorage. In the late 1980s, we began studies tounderstand the nature and mechanism of the acquirement of this unique property upon malignant transformation, by using growth factor-transformable NRK
cells as a model system. The generally held concept
was that anchorage-independent S phase entry is amere consequence of excessive activation of growthsignaling or defects in a growth arrest system, which15

are caused by activation of oncogenes and inactivationof recessive oncogenes. For the past 10 years, westudied genetically and cytologically the properties of
oncogenically stimulated NRK cells and came to theseveral remarkable conclusions: 1) In NRK cells, epidermal growth factor and platelet-derived growth factor signals merge into a common pathway composedof CrkII, Ras and Raf-1, through which many oncogenes induce transformation; 2) Activation of thispathway induces anchorage-independent S phaseentry in a synchronized fashion, but is dispensable forcell proliferation in the presence of anchorage; 3)
Oncogenic stimulation recruits Cdk6 to participate inan essential step of anchorage-independent S phase
onset; 4) Activation of G1 cyclin-dependent kinases(Cdk4/6, Cdk2) is insufficient to induce anchorageindependent S phase onset (15, 16, 32, 39). Recently,
we found that Cdc6, a key protein for the activationof replication origins, requires anchorage or oncogenic
stimulation for its expression, which is regulated at
the levels of transcription and proteolysis via a calpain-like protease, and moreover, that G 1 cyclindependent kinases and Cdc6 constitute key targets forcontrolling the G1-S transition by anchorage and oncogenic signals (Fig. 4)(49).
We believe that this latest finding would provide a16major breakthrough toward the understanding of themechanisms for anchorage loss-invoked restriction of
S phase onset and oncogenically induced anchorageindependent S phase onset, the latter being responsible for the metastatic capability of malignant cells. In
addition, all these findings taken together raise thepossibility that oncogenic transformation is not merely resulted from a failure to arrest due to excessive
growth stimulation or defective cell cycle arrest systems, but from constitutive activation of a dormant
mechanism that is present in normal cells and enables
the cell cycle start in the absence of anchorage (46).

Current Research
We are currently devoting most of our efforts tothe understanding of how Cdc6 expression is regulated by anchorage and oncogenic signals.

Requirement for PI3 kinase in Cdc6 expression
It has been known that phosphoinositol-3 (PI3)kinase is required for the cell cycle start and one of its
targets is p27 cyclin-dependent kinase inhibitor. We
recently found that entirely independent from p27
downregulation, Cdc6 requires PI3 kinase for its
expression during anchorage-dependent, but notanchorage-independent, S phase entry. Treatment ofcells with a specific PI3 kinase inhibitor shuts off
Cdc6 expression despite E2F activation, but interestingly this shutoff is overridden by oncogenic stimulation. Further analysis is underway to identify how PI3
kinase and anchorage signal cooperate to express
Cdc6.

Identification of the protease responsible for theCdc6 degradation and its regulation
The most crucial point in understanding the regulatory mechanism of Cdc6 expression by anchorageand oncogenic stimulation is identification of the
responsible protease and its regulatory mechanism(Fig.4). To this end, a human cdc6 cDNA was isolated,tagged with a histidine tag and transcribed in vitro
and expressed in reticulocyte lysates, and theexpressed His-tagged Cdc6 protein was purified byaffinity chromatography in order to establish an invitro assay system. Analysis of the nature of the putative protease suggests that the cells arrested in G1 byanchorage loss contain the proteolytic activity that
seems to be responsible for Cdc6 degradation.
Analysis of the nature of this protease and its regulation is currently in progress.
Cdk6-cyclin D3 as a highly potent sensitizer ofcells to chemical and physical transformation
We recently found that among D-type cyclin (D1,
D2, D3) and partner kinase (Cdk4, Cdk6) combinations, the Cdk6-D3 complex is unique and can evades
inhibition by p27 and p21, consequently enabling thiscomplex to control proliferation competence under
growth suppressive conditions (47). This finding led
us to investigate the possible effects of the elevatedexpression of this complex on cell’s susceptibility tomalignant transformation, and we have found that 25 fold overexpression of Cdk6-D3 elevates 103-106
folds the susceptibility of rodent fibroblasts to UV
irradiation- or 3-methylcholanthrene-induced malignant transformation. Analysis is in progress to understand this sensitization mechanism.

Future Prospects
There are three key questions regarding malignant
transformation: 1) What is the mechanism by whichtransformed cells acquire the ability to performanchorage-independent S phase onset as a basis formetastatic capability?; 2) What causes the genetic
instability of cancer, which is believed to be a driving
force for malignant progression and what are the key
cell cycle factors activated or inactivated during thisprocess?; 3) What is the mechanism for dedifferentiation of malignant cells? We believe that the series ofour research will certainly offer critical clues to solvethese questions. We particularly believe that the finding that Cdc6, a key factor for the activation of replication origins, absolutely requires anchorage or oncogenic stimulation for its expression is a major breakthrough to understanding the central mechanism of
oncogenically induced anchorage-independent S
phase onset.

Research Grants
1. H. Okayama, Grant-in-Aid for Scientific Research (A),
JSPS, 1992-present. ¥10,000,000 a year.
2. H. Okayama, Grant-in-Aid for Scientific Research on
Priority Areas “Advanced Cancer Research”, MEXT,
1992-1999. ¥32,000,000 a year.
3. H. Okayama, Grant-in-Aid for Scientific Research on
Priority Areas “Cell Cycle Control”, MEXT, 2000-present. ¥60,000,000 a year.

Select Publications
1. Okayama, H., and Berg, P. High efficiency cloning offull length cDNA. Mol. Cell. Biol. 2, 161-170, 1982.
2. Okayama, H., and Berg, P. A cDNA cloning vector
that permits expression of cDNA inserts in mammalian cells. Mol. Cell. Biol. 3, 280-289, 1983.
3. Jolly, D. J., Okayama, H., Berg, P., Esty, A. C., Filpula,D., Bohlen, P., Johnson, G., and Friedman, T.
Isolation and characterization of a full length,
expressible cDNA for human hypoxanthine guanine
phosphoribosyl transferase. Proc. Natl. Acad. Sci.
USA 80, 477-481, 1983.
4. Chin, D. J., Gil, G., Russell, D. W., Liscum, L.,
Luskey, K., Basu, S., Okayama, H., Berg, P.,
Goldstein, J. L., and Brown, M. Nucleotide sequenceof 3-hydroxy-3-methyl-glutaryl coenzyme A reductase, a glycoprotein of endoplasmic reticulum. Nature
308, 613-617, 1984.
5. Yang, Y-C., Okayama, H., and Howley, P. Bovine
papillomavirus contains multiple transforming genes.
Proc. Natl. Acad. Sci. USA 82, 1030-1034, 1985.
6. Okayama, H., Kawaichi, M., Brownstein, M., Lee, F.,
Yokota, T., and Arai, K. High-efficiency cloning offull length cDNA; Construction and screening of
cDNA expression libraries for mammalian cells.
Methods Enz. 154, 3-28, 1987.
7. Chen, C., and Okayama, H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol.
Cell. Biol. 7, 2745-2752, 1987.
8. Okayama, H., and Chen, C. Calcium phosphate mediated gene transfer into established cell lines.
Methods Mol. Biol. pp. 15-21, 1990.
9. Tanaka, K., Miura, N., Satokata, I., Miyamoto, I.,
Yoshida, M. Sato, Y., Kondo, S., Yasui, A., Okayama,
H., and Okada, Y. A cDNA sequence of xeroderma
pigmentosum group A gene encoding a protein withzinc finger structure. Nature 348, 73-76, 1990.

17

10. Inoue, H., Nojima, H., and Okayama, H. High efficiency transformation of Escherichia coli with plasmids. Gene 96, 23-28, 1990.
11. Okazaki, K., Okazaki, N., Kume, K., Jinno, S.,
Tanaka, K., and Okayama. H. High-frequency transformation method and library transducing vectors
for cloning mammalian cDNAs by trans-complementation of Schizosaccharomyces pombe. Nuc. Acids
Res. 18, 6485-6489, 1990.
12. Takamizawa, A., Mori, C., Fuke, I., Manabe, S.,
Murakami, S., Fujita, J., Onishi, E., Andoh, T.,
Yoshida, I., and Okayama, H. Structure and organization of the hepatitis C virus genome isolated fromhuman carriers. J. Virol. 65, 1105-1113, 1991.
13. Nagata, A., Igarashi, M., Jinno, S., Suto, K., andOkayama, H. An additional homolog of the fission
yeast cdc25+ gene occurs in human and is highly
expressed in some cancer cells. New Biologist 3, 959968, 1991.
14. Igarashi, M., Nagata, A., Jinno, S., Suto, K., andOkayama, H. Wee1+-like gene in human cells Nature
353, 80-83, 1991.
15. Masuda, A., Kizaka-Kondoh, S., Miwatani, H.,
Terada, Y., Nojima, H., and Okayama, H. Signaltransduction cascade shared by epidermal growth
factor and platelet-derived growth factor is a major
pathway for oncogenic transformation in NRK cells.
New Biologist 4, 489-503, 1992.
16. Kume, K., Jinno, S., Miwatani, H., Kizaka-Kondoh, S.,
Terada, Y., Nojima, H., and Okayama, H. Oncogenic
signal-induced ability to enter S phase in the absenceof anchorage is the mechanism for the growth oftransformed NRK cells in soft agar. New Biologist 4,
504-511, 1992.
17. Kimura, T., Tanizawa, O., Mori, K., Brownstein, M.,
and Okayama, H. Structure and expression of ahuman oxytocin receptor. Nature 356, 526-529,
1992.
18. Tatsuka, M., Mitsui, H., Wada, M., Nagata, A.,
Nojima, H., and Okayama, H. Elongation factor-1agene determines susceptibility to transformation.
Nature 359, 333-336, 1992.
19. Kizaka-Kondoh, S., Sato, K., Tamura, K., Nojima, H.,
and Okayama, H. Raf-1 protein kinase is an integral
component of the oncogenic signal cascade sharedby epidermal growth factor and platelet-derived
growth factor. Mol. Cell. Biol. 12, 5078-5086, 1992.
20. Tanaka, K., Okazaki, K., Okazaki, N. Ueda, T.
Sugiyama, A., Nojima, H. and Okayama, H. A new
cdc gene required for S phase entry of
Schizosaccharomyces pombe encodes a protein similar to the cdc10+ and SWI4 gene products. EMBO J.
11, 4923-4932, 1992.
21. Benton, B. K., Reid, M. S. and Okayama, H. A
Schizosaccharomyces pombe gene that promotes
sexual differentiation encodes a helix - loop - helix
protein with homology to MyoD. EMBO J. 12, 135143, 1993.
22. Tamura, K., Kanaoka, Y., Jinno, S., Nagata, A., Ogiso,
Y., Shimizu, K., Hayakawa, T., Nojima, H., and18

Okayama, H. Cyclin G: A new mammalian cyclinwith a potential tyrosine phosphorylation site.
Oncogene 8, 2113-2118, 1993.
23. Jinno, S., Suto, K., Nagata, A., Igarashi, M., Kanaoka,
Y., Nojima, H., and Okayama, H. Cdc25A is a novel
phosphatase functioning early in the cell cycle.
EMBO J. 13, 1549-1556, 1994.
24. Sugiyama, A., Tanaka, K., Okazaki, K., Nojima, H.,
and Okayama, H. A zinc finger protein controls theonset of premeiotic DNA synthesis of fission yeast ina Mei2-independent cascade. EMBO J. 13, 1881-1887,
1994.
25. Miyamoto, M., Tanaka, K., and Okayama, H. res2+, a
new member of cdc10+/SWI4 family, controls the∞gstart∞h of mitotic and meiotic cycles in fission
yeast. EMBO J. 13, 1873-1880, 1994.
26. Obara-Ishihara, T., and Okayama, H. A B-type cyclin
regulates conjugation via interaction with cell cycle“start” genes in fission yeast. EMBO J. 13, 1863-1872,
1994.
27. Murakami, H., and Okayama, H. A kinase from fission yeast responsible for blocking mitosis in S
phase. Nature 374, 817-819, 1995.
28. Terada, Y., Tatsuka, M., Jinno, S., and Okayama, H.
Requirement for tyrosine phosphorylation of Cdk4 in
G1 arrest induced by ultraviolet irradiation. Nature
367, 358-362, 1995.
29. Nakashima, N., Tanaka, K., Sturm, S., and Okayama,
H. Fission Yeast Rep2 is a putative transcriptional
activator subunit for the cell cycle “start” function of
Res2-Cdc10. EMBO J. 14, 4794-4802, 1995.
30. Okayama, H., Nagata, A., Jinno, S., Murakami, H.,
Tanaka, K., and Nakashima, N. Cell cycle control infission yeast and mammals identification of new regulatory mechanisms. 1996, Adv. Cancer Re. 69, 1762.
31. Kato, Jr., T., Okazaki, K., Murakami, H., Stettler, S.,
Fantes, A., P., and Okayama, H. Stress signal, mediated by a Hog1-like MAP kinase, controls sexual
development in fission yeast. FEBS Let. 378, 207212, 1996.
32. Kizaka-Kondoh, S., Matsuda, M., and Okayama, H.
CrkII signals from epidermal growth factor receptor
to Ras. Proc. Natl. Acad. Sci. USA 93, 12177-12182,
1996.
33. Yamaguchi, S., Murakami, H., and Okayama, H. A
WD repeat protein controls the cell cycle and differentiation by negatively regulating Cdc2/B-type cyclin
complexes. Mol. Biol. Cell 8, 2475-2486, 1997.
34. Okazaki, N., Okazaki, K., Watanabe, Y., KatoHayashi, M., Yamamoto, M., and Okayama, H. Novel
factor highly conserved among eukaryotes controls
sexual development in fission yeast. Mol. Cell. Biol.
18, 887-895, 1998.
35. Tahara, S., Tanaka, K., Yuasa, Y., and Okayama, H.
Functional domains of Rep2, a transcriptional activator subunit for Res2-Cdc10, controlling the cell cycle
start. Mol. Biol. Cell 9, 1577-1588, 1998.
36. Tsukahara, K., Yamamoto, H., and Okayama, H. An
RNA binding protein negatively controlling differen-tiation in fission yeast. Mol Cell. Biol. 18, 4488-4498,
1998.
37. Jinno, S., Hung, S.-C., and Okayama, H. Cell cycle
start from quiescence controlled by tyrosine phosphorylation of Cdk4. Oncogene 18, 565-577, 1999.
38. Yamamoto, H., Tsukahara, K., Kanaoka, Y., Jinno, S.,
and Okayama, H. Isolation of a mammalian homologue of a fission yeast differentiation regulator. Mol.
Cell. Biol. 19, 3829-3841, 1999.
39. Jinno, S., Hung, S.-C., Yamamoto, Y., Lin, J., Nagata,A., and Okayama, H. Oncogenic stimulation recruits
Cdk6 in the cell cycle start in rat fibroblast. Proc.
Natl. Acad. Sci. USA 96, 13197-13202, 1999.
40. Suto, K., Murakami, H., Nagata, A., and Okayama, H.
A double-strand Break repair component is essential
for S phase completion in fission yeast cell cycling.
Mol. Biol. Cell 10, 3331-3343, 1999.
41. Masuda, M., Nagai, Y., Oshima, N., Tanaka, K.,
Murakami, H., Igarashi, H., and Okayama H. Genetic
Studies with the Fission Yeast Schizosaccharomyces
pombe Suggest Involvement of Wee1, Ppa2, andRad24 in Induction of Cell Cycle Arrest by Human
Immunodeficiency Virus Type 1 Vpr. J. Virol. 74,
2636-2646, 2000.
42. Tanaka, K., Yonekawa, T., Kawasaki, Y., Kai, M.,
Furuya, K., Iwasaki, M., Murakami, H., Yanagida, M.,
and Okayama, H. Fission yeast Eso1p is required forestablishing chromatid cohesion during S phase.
Mol. Cell. Biol. 20, 3459-3469, 2000.
43. Yamaguchi, S., Okayama, H. and Nurse, P. Fission
yeast Fizzy-related protein Srw1p is a G(1)-specificpromoter of mitotic cyclin B degradation. EMBO J.
19, 3968-3977, 2000.
44. Tanaka, K., and Okayama, H. A Pcl-like cyclin activates the Res2p-Cdc10p cell cycle `start’ transcriptional factor in fission yeast. Mol. Biol. Cell 11, 28452862, 2000.
45. Nakamura, E., Isobe, H., Tomita, N., Sawamura, M.,
Jinno, S., and Okayama, H. Funtionalized fullerene
as an artificial vector for transfection. Angew. Chem.
112, 4424-4427, 2000.
46. Jinno, S., Lin, J., Yageta, M., and Okayama, H.
Oncogenic cell cycle start control. Mut. Res. 477, 2329, 2001.
47. Lin, J., Jinno, S., and Okayama H. Cyclin D3-Cdk6
complex evades inhibition by inhibitor proteins anduniquely controls cell’s growth competence.
Oncogene 20, 2000-2009, 2001.
48. Tanaka, K., Hao, Z., Kai, M. and Okayama, H.
Establishment and maintenance of sister chromatid
cohesion in fission yeast by a unique mechanism.
EMBO J. 20, 5779-5790, 2001.
49. Jinno, S., Yageta, M., Nagata, A. and Okayama, H.
Cdc6 requires anchorage for its expression.
Oncogene 21, 1777-1784, 2002.
50. Hiroi, N., Jinno, S., and Okayama, H. Mammalian
Rcd1 is a novel transcriptional cofactor that mediates
retinoic acid-induced cell differentiation. EMBO J. 21,
5235-5244, 2002.

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Department of
Cellular Signaling
Outline and Research Objectives
When cells are stimulated, by the serial actions of various enzymes, biologically active
lipid molecules are produced and released. They include prostaglandins, leukotrienes,
platelet-activating factor (PAF), lysophosphatidic acid (LPA) etc., collectively termed lipid
mediators (Fig 1). In concert with various neurotransmitters and hormones, these lipid
mediators are playing important roles in self-defenses and maintenance of homeostasis.
Unlike the biogenic amines or peptide mediators, lipid mediators are not stored in granules,
but they are biosynthesized from precursor lipids when necessary. To elucidate the functions of lipid mediators, we isolated enzymes involved in the biosyntheses and metabolismof lipid mediators, isolated G-protein-coupled receptors, and identification of intracellularsignaling. We are especially interested in identifying the specific roles of lipid mediators in
inflammatory responses as well as neuronal functions. We generated several transgenic orknockout mice and analysed their roles in vivo. By mass spectrometric analyses, we alsomeasured dynamic changes in membrane lipid compositions, the significance of thesechanges being in general signaling pathway. In the postsequece of whole human genome,
the intensive studies of protein regulation as well as behavior of small lipid metabolites areof the highest importance.

Faculties and Students
Professor
Takao Shimizu, M.D., Ph.D. (1991-)
Associate Professor Takehiko Yokomizo, M.D., Ph.D.
Associate .................................3
Postdoctoral Fellows...............4
Graduate Students ................12
Research Students...................7
Secretary .................................2

Past Research and Major Accomplishments
We have purified and cDNA cloned various

enzymes involved in the biosyntheses and metabolism of lipid mediators. Leukotriene A4 hydrolase,
one of the key enzyme in the biosynthesis of chemotactic leukotrienes, was cloned and characterized in
our laboratory. This achievement was the first successful example of molecular cloning of the enzymes
in the field of lipid mediators. We also obtained PAF
receptor, as the first successful example of receptor
cloning of lipid mediators, followed by cloning receptors of various lipid mediators. We determined theregulation of these enzymes by Ca-dependent intracellular translocation, phosphorylation by various kinases, and other posttranslational modifications. We also

Membrane phospholipids

20

Phospholipids:

Lyso-phospholipids:

Eicosanoids:

Platelet-activating factor
Psychosine
Sphingophosphocholine

Lyso-phosphatidylcholine
Lyso-phosphatidic acid
Sphigosine 1-phosphate
2-Arachidonoyl-glycerol

Prostaglandins
Thromboxanes
Leukotrienes
Lipoxinsgenerated PAF receptor-overexpressing mice, PAF
receptor-deficient mice, leukotriene receptor-deficient
mice, and the mice deficient in cytosolic phospholipase A2 (cPLA2), which are involved in the synthesesof eicosanoids and PAF. By analyzing the phenotypesof these mice, we reported that the mediators are
involved in the pathogenesis of various inflammatory
and immune disorders, as well as normal physiology
including reproduction and synaptic plasticity.
Independently, one of the Associate is working onaxon guidance molecules, by establishing knockout
mice of guidance molecules (Sema 3A), and analysesof their multiple phenotypes.

Current Research
Followings are ongoing projects in our laboratory.
1. Elucidation of enzymes in the biosyntheses oflipid mediators, their regulation and mechanismof intracellular translocation by stimuli.
2. Deorphaning projects of putative lipid mediator
GPCRs, which include indentificaion of novel lipid
mediators (Fig. 2).
3. Elucidation of GPCR-sorting mechanism in polarized cells (kidney epithelial cells, neuron etc.)
4. Phenotype analyses of various genetic engineered
mice. Understanding of the molecular mechanismof individual phenotypes in depth.

21

5. Establishment of novel gene-targeted mice(enzymes and receptors), and their congenic line
(B6, Balb/c, DBA etc.)
6. Identification of Sema3A knockout mice, in nervous systems, olfactory systems, bone formation.

3. 1999-2002 Grant from the Ministry of Education,
Science, Culture, Sports, and Technology of Japan (A,B, and C)
4. 1997 Priority Area A, from the Ministry of
Education, Science, Culture, Sports, and Technologyof Japan.
5. 1996-2000 Ministry of Welfare, Health and Labor.

Future Prospects
We will pursue following different projects, leadby each faculty member.
1. Identification of the roles of lipid mediators in thecentral nervous system. For this purpose, we will
analyze the localization and subcellular localization of enzymes involved in the biosyntheses oflipid mediators. It is the most important to determine, how and which direction (either axon or
dendrite), these enzyme cause translocation in a
stimulus-dependent manner. The mice deficient of
enzymes or receptors of lipid mediators are useful
to identify the roles of mediators in the central
nervous systems. Some link has been reported
between Sema3A axon guidance molecules andlipid mediators. By the use of double knockout
mice (Sema3A and lipid mediators), more confirmative and direct evidence will be obtained.
2. We have so far obtained various phenotypes ofgenetic engineered mice, but molecular mechanisms underlying these phenotypes remain mostly
unknown. The link between lipid mediators andhormones, neurotransmitters, and cytokines will
be analyzed to obtain a whole view.
3. By mass spectrometric analyses, we like to findout the temporal and positional dynamic changesof membrane compositions including Raft andcaveolae. Also, mass spectrometry attached with
HPLC will aid in the identification of novel natural
lipid mediators (deorphaning project). For these
purposes, we recently built a new research groupcalled Department of Lipid Metabolome, by donation. We have collected several excellent lipid biochemists, and mass spectrometer specialists.
Metabolome (metabolomics) is a new researchstrategy after genome and proteome research. The
study is to analyze in detail small-sized molecules(metabolites), and in combination with proteomics, we will gain a more broad view how cells
adapt to a new environment, change membrane
fluidity, and cause metabolic changes, which finally yield to gene expression.

Research Grants
1. 1998-2003 CREST of Japan Science and Technology
Corporation
2. 2001-2004 PREST of Japan Science and Technology
Corporation22

Select Publications (50 among 128 publications between 1991 and 2002)
1. Ito, N., Yokomizo, T., Sasaki, T., Kurosu, H.,
Penninger, J., Kanaho, Y., Katada, T., Hanaoka, K.,
and Shimizu, T. Requirement of phosphatidylinositol
3-kinase-activation and Ca influx for leukotriene B4induced enzyme release. J. Biol. Chem. in press,
2002.
2. Stromgaard, K., Saito, D.R., Shindou, H., Ishii, S.,
Shimizu, T., and Nakanishi, K. Ginkgolide derivatives for photolabeling studies: preparation and pharmacological evaluation. J. Med. Chem. 45,4038-46,
2002.
3. Wong, D.A., Uozumi, N., Kita, Y., and Shimizu, T.
Discrete role for cytosolic phospholipase A2-alpha inplatelets: Studies using single and double mutant
mice of cytosolic and group IIA-secretory phospholipase A2. J. Exp. Med. 196,349-57, 2002.
4. Ogasawara, H., Ishii, S., Yokomizo, T., Kakinuma, T.,
Komine, M., Tamaki, K., Shimizu, T., and Izumi, T.
Characterization of mouse cysteinyl leukotriene
receptors, mCysLT1 and mCysLT2: Differential
pharamacological properties and tissue distribution.
J. Biol. Chem. 277, 18763-18768, 2002.
5. Nagase, T., Uozumi, N., Ishii, S., Kita, Y., Yamamoto,H., Ohga, E., Ouchi, Y. and Shimizu, T. A pivotal roleof cytosolic phospholipase A2 in bleomycin-induced
pulmonary fibrosis. Nature Med. 8, 480-484, 2002.
6. Ohshima, N., Ishii, S., Izumi, T., and Shimizu, T.
Receptor-dependent metabolism of platelet-activating factor in murine macrophages. J. Biol. Chem. 277,9722-9727, 2002.
7. Nagase, T., Ishii, S., Shindou, H., Ouchi, Y., andShimizu, T. Airway hyperresponsiveness in transgenic mice overexpressing platelet activating factor
receptor is mediated by an atropine-sensitive pathway. Am. J. Respir. Crit. Care Med. 165, 200-205,
2002.
8. Yokomizo, T., Izumi, T., and Shimizu, T. Leukotriene
B4: metabolism and signal transduction. Arch.
Biochem. Biophys. 385, 231-241, 2001.
9. Yokomizo, T., Kato, K., Hagiya, H., Izumi, T., andShimizu, T. Hydroxyeicosanoids bind to and activate
the low affinity leukotriene B4 receptor, BLT2. J.
Biol. Chem. 276, 12454-12459, 2001.
10. Wu, C., Stojanov, T., Chami, O., Ishii, S., Shimizu, T.,
Li, A., and O’Neill, C. Evidence for the autocrine
induction of capacitation of mammalian spermatozoa. J. Biol. Chem. 276, 26962-26968, 2001.
11. Yamashita, A., Kawagishi, N., Miyashita, T.,
Nagatsuka, T., Sugiura, T., Kume, K., Shimizu, T.,and Waku, K. ATP-independent fatty acyl-coenzyme
A synthesis from phospholipid: coenzyme A-dependent transacylation activity toward lysophosphatidic
acid catalyzed by acyl-coenzyme A:lysophosphatidic
acid acyltransferase. J. Biol. Chem. 276, 26745-26752,
2001.
12. Kim, Y. J., Noguchi, S., Hayashi, Y. K., Tsukahara, T.,
Shimizu, T., and Arahata, K. The product of an oculopharyngeal muscular dystrophy gene, poly(A)-binding protein 2, interacts with SKIP and stimulates
muscle-specific gene expression. Hum. Mol. Genet.
10, 1129-1139, 2001.
13. Fukunaga, K., Ishii, S., Asano, K., Yokomizo, T.,
Shiomi, T., Shimizu, T., and Yamaguchi, K. Single
nucleotide polymorphism of human platelet-activating factor receptor impairs G-protein activation. J.
Biol. Chem. 276, 43025-43030, 2001.
14. Aihara, M., Ishii, S., Kume, K., and Shimizu, T.
Interaction between neurone and microglia mediatedby platelet-activating factor. Genes Cells 5, 397-406,
2000.
15. Noiri, E., Yokomizo, T., Nakao, A., Izumi, T., Fujita,T., Kimura, S., and Shimizu, T. An in vivo approach
showing the chemotactic activity of leukotriene B(4)in acute renal ischemic-reperfusion injury. Proc. Natl.
Acad. Sci. USA 97, 823-828, 2000.
16. Nagase, T., Uozumi, N., Ishii, S., Kume, K., Izumi, T.,
Ouchi, Y., and Shimizu, T. Acute lung injury by sepsis and acid aspiration: a key role for cytosolic phospholipase A2. Nat. Immunol. 1, 42-46, 2000.
17. Kato, K., Yokomizo, T., Izumi, T., and Shimizu, T.
Cell-specific transcriptional regulation of humanleukotriene B(4) receptor gene. J. Exp. Med. 192, 413420, 2000.
18. Yokomizo, T., Kato, K., Terawaki, K., Izumi, T., andShimizu, T. A second leukotriene B(4) receptor,
BLT2. A new therapeutic target in inflammation andimmunological disorders. J. Exp. Med. 192, 421-432,
2000.
19. Takaku, K., Sonoshita, M., Sasaki, N., Uozumi, N.,
Doi, Y., Shimizu, T., and Taketo, M. M. Suppressionof intestinal polyposis in Apc(delta 716) knockout
mice by an additional mutation in the cytosolic phospholipase A(2) gene. J. Biol. Chem. 275, 3401334016, 2000.
20. Nakatani, N., Uozumi, N., Kume, K., Murakami, M.,
Kudo, I., and Shimizu, T. Role of cytosolic phospholipase A2 in the production of lipid mediators and histamine release in mouse bone-marrow-derived mast
cells. Biochem. J. 352 Pt 2, 311-317, 2000.
21. Okamoto, H., Takuwa, N., Yokomizo, T., Sugimoto,N., Sakurada, S., Shigematsu, H., and Takuwa, Y.
Inhibitory regulation of Rac activation, membrane
ruffling, and cell migration by the G protein-coupledsphingosine-1-phosphate receptor EDG5 but not
EDG1 or EDG3. Mol. Cell Biol. 20, 9247-9261, 2000.
22. Kobayashi, K., Ishii, S., Kume, K., Takahashi, T.,
Shimizu, T., and Manabe, T. Platelet-activating factor
receptor is not required for long-term potentiation in

the hippocampal CA1 region. Eur. J. Neurosci. 11,
1313-1316, 1999.
23. Nagase, T., Ishii, S., Kume, K., Uozumi, N., Izumi, T.,
Ouchi, Y., and Shimizu, T. Platelet-activating factor
mediates acid-induced lung injury in genetically engineered mice. J. Clin. Invest. 104, 1071-1076, 1999.
24. Hirabayashi, T., Kume, K., Hirose, K., Yokomizo, T.,
Iino, M., Itoh, H., and Shimizu, T. Critical duration of
intracellular Ca2+ response required for continuoustranslocation and activation of cytosolic phospholipase A2. J. Biol. Chem. 274, 5163-5169, 1999.
25. Ishii, I., Saito, E., Izumi, T., Ui, M., and Shimizu, T.
Agonist-induced sequestration, recycling, and resensitization of platelet-activating factor receptor. Roleof cytoplasmic tail phosphorylation in each process.
J. Biol. Chem. 273, 9878-9885, 1998.
26. Hoshino, M., Izumi, T., and Shimizu, T. Leukotriene
D4 activates mitogen-activated protein kinasethrough a protein kinase Calpha-Raf-1-dependent
pathway in human monocytic leukemia THP-1 cells.
J. Biol. Chem. 273, 4878-4882, 1998.
27. Ishii, S., Kuwaki, T., Nagase, T., Maki, K., Tashiro,F., Sunaga, S., Cao, W. H., Kume, K., Fukuchi, Y.,
Ikuta, K., Miyazaki, J., Kumada, M., and Shimizu, T.
Impaired anaphylactic responses with intact sensitivity to endotoxin in mice lacking a platelet-activating
factor receptor. J. Exp. Med. 187, 1779-1788, 1998.
28. Hirohashi, T., Suzuki, H., Ito, K., Ogawa, K., Kume,
K., Shimizu, T., and Sugiyama, Y. Hepatic expressionof multidrug resistance-associated protein-like proteins maintained in eisai hyperbilirubinemic rats.
Mol. Pharmacol. 53, 1068-1075, 1998.
29. Ishii, S., Nagase, T., Tashiro, F., Ikuta, K., Sato, S.,
Waga, I., Kume, K., Miyazaki, J., and Shimizu, T.
Bronchial hyperreactivity, increased endotoxin
lethality and melanocytic tumorigenesis in transgenic mice overexpressing platelet-activating factor
receptor. EMBO. J. 16, 133-142, 1997.
30. Kume, K., and Shimizu, T. Platelet-activating factor(PAF) induces growth stimulation, inhibition, andsuppression of oncogenic transformation in NRK
cells overexpressing the PAF receptor. J. Biol. Chem.
272, 22898-22904, 1997.
31. Ishii, I., Izumi, T., Tsukamoto, H., Umeyama, H., Ui,
M., and Shimizu, T. Alanine exchanges of polaramino acids in the transmembrane domains of aplatelet-activating factor receptor generate both constitutively active and inactive mutants. J. Biol. Chem.
272, 7846-7854, 1997.
32. Honda, Z., Suzuki, T., Hirose, N., Aihara, M.,
Shimizu, T., Nada, S., Okada, M., Ra, C., Morita, Y.,
and Ito, K. Roles of C-terminal Src kinase in the initiation and the termination of the high affinity IgE
receptor-mediated signaling. J. Biol. Chem. 272,
25753-25760, 1997.
33. Yokomizo, T., Izumi, T., Chang, K., Takuwa, Y., andShimizu, T. A G-protein-coupled receptor forleukotriene B4 that mediates chemotaxis. Nature
387, 620-624, 1997.

23

34. Uozumi, N., Kume, K., Nagase, T., Nakatani, N.,
Ishii, S., Tashiro, F., Komagata, Y., Maki, K., Ikuta,
K., Ouchi, Y., Miyazaki, J., and Shimizu, T. Role ofcytosolic phospholipase A2 in allergic response andparturition. Nature 390, 618-622, 1997.
35. Yokomizo, T., Ogawa, Y., Uozumi, N., Kume, K.,
Izumi, T., and Shimizu, T. cDNA cloning, expression,and mutagenesis study of leukotriene B4 12-hydroxydehydrogenase. J. Biol. Chem. 271, 2844-2850,
1996.
36. Ferby, I. M., Waga, I., Hoshino, M., Kume, K., andShimizu, T. Wortmannin inhibits mitogen-activatedprotein kinase activation by platelet-activating factor
through a mechanism independent of p85/p110-type
phosphatidylinositol 3-kinase. J. Biol. Chem. 271,
11684-11688, 1996.
37. Kishimoto, S., Shimadzu, W., Izumi, T., Shimizu, T.,
Fukuda, T., Makino, S., Sugiura, T., and Waku, K.
Regulation by IL-5 of expression of functional
platelet-activating factor receptors on humaneosinophils. J. Immunol. 157, 4126-4132, 1996.
38. Mori, M., Aihara, M., Kume, K., Hamanoue, M.,
Kohsaka, S., and Shimizu, T. Predominant expression of platelet-activating factor receptor in the rat
brain microglia. J. Neurosci. 16, 3590-3600, 1996.
39. Mutoh, H., Fukuda, T., Kitamaoto, T., Masushige, S.,
Sasaki, H., Shimizu, T., and Kato, S. Tissue-specificresponse of the human platelet-activating factor
receptor gene to retinoic acid and thyroid hormone
by alternative promoter usage. Proc. Natl. Acad. Sci.
USA 93, 774-779, 1996.
40. Watanabe, T., Waga, I., Honda, Z., Kurokawa, K.,
and Shimizu, T. Prostaglandin F2 alpha stimulates
formation of p21ras-GTP complex and mitogen-activated protein kinase in NIH-3T3 cells via Gq-proteincoupled pathway. J. Biol. Chem. 270, 8984-8990,
1995.
41. Honda, Z., Takano, T., Hirose, N., Suzuki, T., Muto,
A., Kume, S., Mikoshiba, K., Itoh, K., and Shimizu, T.
Gq pathway desensitizes chemotactic receptorinduced calcium signaling via inositol trisphosphate
receptor down-regulation. J. Biol. Chem. 270, 48404844, 1995.
42. Shirasaki, H., Nishikawa, M., Adcock, I. M., Mak, J.
C., Sakamoto, T., Shimizu, T., and Barnes, P. J.
Expression of platelet-activating factor receptor
mRNA in human and guinea pig lung. Am. J. Respir.
Cell Mol. Biol. 10, 533-537, 1994.
43. Takano, T., Honda, Z., Sakanaka, C., Izumi, T.,
Kameyama, K., Haga, K., Haga, T., Kurokawa, K., andShimizu, T. Role of cytoplasmic tail phosphorylationsites of platelet-activating factor receptor in agonistinduced desensitization. J. Biol. Chem. 269, 2245322458, 1994.
44. Honda, Z., Takano, T., Gotoh, Y., Nishida, E., Ito, K.,
and Shimizu, T. Transfected platelet-activating factor
receptor activates mitogen-activated protein (MAP)kinase and MAP kinase kinase in Chinese hamster
ovary cells. J. Biol. Chem. 269, 2307-2315, 1994.

24

45. Ferby, I. M., Waga, I., Sakanaka, C., Kume, K., andShimizu, T. Wortmannin inhibits mitogen-activatedprotein kinase activation induced by platelet-activating factor in guinea pig neutrophils. J. Biol. Chem.
269, 30485-30488, 1994.
46. Bito, H., Mori, M., Sakanaka, C., Takano, T., Honda,
Z., Gotoh, Y., Nishida, E., and Shimizu, T. Functional
coupling of SSTR4, a major hippocampal somatostatin receptor, to adenylate cyclase inhibition,arachidonate release and activation of the mitogenactivated protein kinase cascade. J. Biol. Chem. 269,
12722-12730, 1994.
47. Bito, H., Nakamura, M., Honda, Z., Izumi, T.,
Iwatsubo, T., Seyama, Y., Ogura, A., Kudo, Y., andShimizu, T. Platelet-activating factor (PAF) receptor
in rat brain: PAF mobilizes intracellular Ca2+ in hippocampal neurons. Neuron 9, 285-294, 1992.
48. Nakamura, M., Honda, Z., Izumi, T., Sakanaka, C.,
Mutoh, H., Minami, M., Bito, H., Seyama, Y.,
Matsumoto, T., Noma, M., and Shimizu, T.
Molecular cloning and expression of platelet-activating factor receptor from human leukocytes. J. Biol.
Chem. 266, 20400-20405, 1991.
49. Honda, Z., Nakamura, M., Miki, I., Minami, M.,
Watanabe, T., Seyama, Y., Okado, H., Toh, H., Ito,K., Miyamoto, T., and Shimizu, T. Cloning by functional expression of platelet-activating factor receptor from guinea-pig lung. Nature 349, 342-346, 1991.
50. Watanabe, T., Yatomi, Y., Sunaga, S., Miki, I., Ishii,A., Nakao, A., Higashihara, M., Seyama, Y., Ogura,
M., Saito, H., Kurokawa, K., and Shimizu, T.
Characterization of prostaglandin and thromboxane
receptors expressed on a megakaryoblastic leukemia
cell line, MEG-01s. Blood 78, 2328-2336, 1991.

Department of Physiological
Chemistry and Metabolism
Outline and Research Objectives
The Department of Physiological Chemistry and Nutrition, the predecessor of the present department, was founded in 1952. The successive professors were as follows;
The first professor:
Haruhisa Yoshikawa (1952~1969)
The second professor: Osamu Hayaishi (1970~1974)
The third professor: Yoshitake Mano (1974~1982)
The fourth professor: Yousuke Seyama (1983~2001)
During the past 50 years, these professors lead high-quality researches in the field of biochemistry, metabolism and nutrition and greatly contributed to scientific progresses inJapan. Upon the restructuring of the university system in 1997, the department was
renamed ‘Department of Physiological Chemistry and Metabolism’ as one unit of theSpecialty of Molecular Cell Biology. In 2002, Dr. Hiroki Kurihara was designated as a successor of Professor Yousuke Seyama, and started new researches on developmental biology andtechnology as well as regenerative medicine.

Faculties and Students
Professor and Chair Hiroki Kurihara, M.D., Ph.D. (2002~)
Associate .................................3
Postdoctoral Fellow ................1
Graduate Student....................2
Secretary .................................1

Past Research and Major Accomplishments
1. Vascular Biology(1) Biology of Vasoactive Peptidesi) Endothelin
a. Discovery and Basic Characterization ofEndothelin
Our researches started with contribution to thediscovery of endothelin (ET-1), an endotheliumderived vasoactive peptide, with Drs. Masashi
Yanagisawa, Tomoh Masaki and colleagues in
1987. Then, we studied the role of ET-1 in vascularphysiology and pathophysiology and revealed its
vasoconstrictive effect on in vivo canine coronary
arteries and induction of the ET-1 gene by somecytokines and flow-induced mechanical stress invascular endothelial cells.
b. Endothelin-1 Knockout Mice
In 1991, we introduced the technique of genetargeting in mice to our lab to further analyze thebiological implication of ET-1. As a result, wefound two novel and important roles of ET-1. First,
ET-1 proved to lower blood pressure when it actsin the central nervous system although it mainly
acts as a pressor in the periphery. ET-1 also modulates stress responses in the central nervous system by regulating catecholamine metabolism.
Second, ET-1 proved to be essential for embryogenesis. The phenotype of mice lacking ET-1
involved craniofacial and cardiac neural crestderived structures including the branchial archesand great vessels. Indeed, ET-1 was the firstvasoactive substance bound to G protein-coupled
receptors that proved to be involved in embryonicdevelopment (See 2-1)).
c. Endothelin-1 Overexpressing MiceWe found that the expression of ET-1 and its
processing enzyme ECE-1 is increased in the lesionof human atherosclerosis and experimental vascular injury. The effect of overexpressed ET-1 wasfurther analyzed in transgenic mice and proved tocause mesangial hyperplasia and glomerulosclerosis in the kidney. This result was indicative of thestimulating effect of ET-1 on cellular proliferation
and remodeling in vivo.
ii) Adrenomedullin
a. Adrenomedullin Overexpressing Mice
Adrenomedullin (AM), a vasodilating peptide,can be regarded as a counterpart of ET-1 in vascular tone regulation. We investigated the in vivo
effect of AM by producing transgenic mice overexpressing AM and found NO-dependent hypotension and resistance to endotoxin shock (decreasein lethality) in transgenic mice. These results suggested that AM may lower blood pressure by stim-

25

ulating NO production and that AM may protect
tissues and organs from shock-induced damage.
b. Adrenomedullin Knockout Mice
The biological importance of AM was further
investigated by gene targeting. We demonstrated
that AM is indispensable for the vascular morphogenesis during embryonic development and for
postnatal regulation of blood pressure by stimulating NO production, confirming the hypothesis
derived from AM overexpression mice.
iii) CGRPCGRP is structural related to AM and shares thecommon receptor CRLR. To investigate systematically the differential role of the AM/CGRP family
members, we established alphaCGRP knockout
mice. The resultant phenotype revealed thatalphaCGRP contributes to the regulation of cardiovascular function through inhibitory modulation of
sympathetic nervous activity and that AM and
CGRP have distinct physiological roles.
(2) Transgenic techniques
During gene manipulation studies in mice, we
found that the ET-1 gene promoter is useful for vesselselective gene expression. This promoter was used
not only to overexpress ET-1 and AM but also tomake endothelial NO synthase and 15-lipoxygenase
overexpressing mice by our collaborators with fruitfulresults. Recently, we realized targeting gene expression in vascular smooth muscle cells by using the ETA receptor (ETAR) gene promoter.
2. Developmental Biology(1) Neural Crest Development
Craniofacial and cardiovascular defects in ET-1
knockout embryos gave a clue to the elucidation of
mechanisms for branchial arch formation contributedby neural crest cells. By subsequent analysis, werevealed that ET-1 regulates several downstream
genes such as HAND1/2 and Goosecoid transcription
factors to contribute to cranial/cardiac neural crest
development. Recently, we established ETAR promoter::GFP and ETAR promoter::TVA (avian retrovirus
receptor) to visualize and isolate ETAR+ neural crest
cells and to transfer genes specifically to them via
retroviral vectors. These systems serve as very usefultools for the analysis of neural crest-derived branchialarch development.
(2) Vascular Development
ET-1 knockout mice also demonstrated defects in
the great vessels and cardiac outflow tract, to which
cardiac neural crest cells largely contribute. From thisresult, we found that ET-1 is important in vascularsmooth muscle cell development originated from neu-

26

ral crest cells and that the ET-1to HAND2 signaling
pathway seems to be critical. The outcome from AM
knockout mice showed that AM is important in theformation of basement membrane during angiogenesis to stabilize vascular network.
3. Collaborative Works(1) Biology of Metalloproteinase
In collaboration with Dr. Kouji Matsushima (Dept.
of Molecular Preventive Medicine), we exploited thebiological implication of ADAMTS-1, a member of the
ADAM-type metalloproteinase family, by gene targeting. We found that ADAMTS-1 is important for normal growth, fertility and organ developemnt (the kidneys, adrenal glands, adipose tissues etc.).
(2) Biology of Antimicrobial Peptides
In collaboration with Drs. Yasuyoshi Ouchi and
Takahide Nagase (Dept. of Geriatrics), we are studying
about defensins, endogenous antimicrobial peptides.
We have identified several novel types of human andmouse beta-defensins which are expressed skeletal
muscle and epididymis.

Current Research
1. Vascular Biology
We have converged our previous achievement ofvascular biology on the aspect of developmental biology including vascular development as described
below.
2. Developmental Biology(1) Neural Crest Development
Extending our achievement of ET-1 knockout
mice, we are studying the mechanism how the intercellular and intracellular signaling system related ET-1
is involved in neural crest development and branchialarch formation because it may give a clue to theinsight how signaling interactions can change cellularbehavior to lead to morphogenesis. For this purpose,
we have established GFP and TVA transgenic mice as
described and are now establishing some other mice
which will be useful for the analysis of branchial archformation. We are also performing a systematic
screening for differential gene expression by DNA
microarray followed by in situ hybridization. By thismethod, several important genes are to be obtained.
We are also studying about neural crest differentiation using murine neural crest cell culture and Sendai
virus vectors developed by Drs. Yoshiyuki Nagai andAtsushi Kato. In our preliminary result, we realized
nearly 100 % gene transfer into neural crest cells andare analyzing the effect of some genes on neural crestdifferentiation. Using some markers, we are also trying to isolate neural crest stem cells.
We are further looking for the mechanism how
some transcription factors important for neural crest
development can exert their effects. For this purpose,
we are performing yeast two-hybrid screening using
Pax3 and HAND2 as baits. We have obtained several
interesting clones and are analyzing their molecularcharacteristics.
(2) Vascular Development
Above-mentioned studies concerning neural crest
differentiation include the issue of vascular smoothmuscle cell differentiation and we are trying to identify the target genes of the ET-1-HAND2 pathway thatis important for smooth muscle differentiation. Twohybrid studies also involve this theme. Abnormalities
in vascular formation in AM knockout mice are also
being investigated further concerning the mechanism.
In addition, we have started studies on vascularendothelial cell differentiation using the embryonicstem cell system and are looking for possible interaction between neural crest-derived smooth muscle precursors and endothelial precursors. It may clarify anovel cellular interaction mechanism which can stimulate vascular formation.
3. Collaborative Works
In collaboration with Drs. Yasuyoshi Ouchi and
Takahide Nagase (Dept. of Geriatrics), we are starting
further study on defensins using gene manipulationin mice.

Future Prospects(1) Basic research
Our future study will focus mainly on neural crestand vascular development. Especially, major questionsare (1) how do intracellular mechanisms modulate
stem cell behavior including fate determination anddifferentiation, and (2) how are cell behaviors integrated into morphogenesis and organogenesis. By
studying neural crest development as a model, we
expect that we can approach these general issues. For
our coming research, we are introducing some newtechniques such as nuclear transfer.
(2) Applied research
Our current researches on vascular developmentusing embryonic stem cells are close to appliedresearches in the field of regenerative medicine. On
this background together with nuclear transfer technique, we are planning translational researches togenerate vessels and other tissues and organs related

neural crest cells.

Research Grants
1. JSPS (Japan Society for the Promotion of Science)
Research for the Future Program (2000.4~2004.3)
total 240,000,000 yen.
2. Grants-in-Aid for Scientific Research from JSPS,
Scientific Research (B)(2002.4~2004.3) total 14,900,000 yen
3. Grants-in-Aid for Scientific Research from JSPS,
Exploratory Research (C)(2002.4~2004.3) total 9,900,000 yen
4. Grants-in-Aid for Scientific Research from the
Ministry of Education, Science and Culture, Japan,
Priority Areas Research (A), Allotted.
(2000.4~2001.3) total 1,800,000 yen.
5. the Research Grant for Cardiovascular Diseases (14C1) from the Ministry of Health and Welfare(2002.4~2005.3) total 6,300,000 yen.

Select Publications
1. Yanagisawa M, Kurihara H, Kimura S, Tomobe Y,
Kobayashi M, Mitsui Y, Yazaki Y, Goto K, Masaki T.
A novel vasoconstrictor peptide produced by vascular endothelial cells. Nature 332: 411-415, 1988.
2. Komuro I, Kurihara H, Sugiyama T, Yoshizumi M,
Takaku F, Yazaki Y. Endothelin stimulates c-fos andc-myc expression and proliferation of vascularsmooth muscle cells. FEBS Lett. 238: 249-252, 1988.
3. Kurihara H, Yoshizumi M, Sugiyama T, Takaku F,
Yanagisawa M, Masaki T, Hamaoki M, Kato H,
Yazaki Y. Transforming growth factor-ß stimulates
the expression of endothelin mRNA by vascularendothelial cells. Biochem. Biophys. Res. Commun.
359: 1435-1440, 1989.
4. Kurihara H, Yoshizumi M, Sugiyama T, Yamaoki K,
Nagai R, Takaku F, Satoh H, Inui J, Yanagisawa M,
Masaki T, Yazaki Y. The possible role of endothelin1 in the pathogenesis of coronary vasospasm. J.
Cardiovasc. Pharmacol. 13(Suppl. 5): S132-S137,
1989.
5. Kurihara H, Yamaoki K, Nagai R, Yoshizumi M,
Takaku F, Satoh H, Inui J, Yazaki Y. Endothelin: apotent vasoconstrictor associated with coronary
vasospasm. Life Sci. 44: 1937-1943, 1989.
6. Goto K, Kasuya Y, Matsuki N, Takuwa Y, Kurihara H,
Ishikawa T, Kimura S, Yanagisawa M, Masaki T.
Endothelin activates the dihydropyridine-sensitive,
voltage-dependent Ca2+-channel in vascular smoothmuscle. Proc. Natl. Acad. Sci. U.S.A. 86: 3915-3918,
1989.
7. Ouchi Y, Kim S, Souza AC, Iijima S, Hattori A, Orimo
H, Yoshizumi M, Kurihara H, Yazaki Y. Central effectof endothelin on blood pressure in conscious rats.
Am J. Physiol. 256: H1747-H1751, 1989.
8. Yoshizumi M, Kurihara H, Sugiyama T, Takaku F,
Yanagisawa M, Masaki T, Yazaki Y. Hemodynamic

27

shear stress stimulates endothelin production by cultured endothelial cells. Biochem. Biophys. Res
Commun. 161: 859-864, 1989.
9. Yoshizumi M, Kurihara H, Morita T, Yamashita T,
Oh-hashi Y, Sugiyama T, Takaku F, Yanagisawa M,
Masaki T, Yazaki Y. Interleukin-1 induces the production of endothelin-1 by cultured endothelial cells.
Biohem. Biophys. Res. Commun. 166: 324-329, 1990.
10. Morita T, Kurihara H, Yoshizumi M, Maemura K,
Sugiyama T, Nagai R, Yazaki Y. Polymorphonuclear
leukocytes have dual effects on endothelin-1: theinduction of endothelin-1 mRNA expression in vascular endothelial cells and modification of theendothelin-1 molecule. Heart Vessels 8: 1-6, 1993.
11. Morita T, Kurihara H, Maemura K, Yoshizumi M,
Yazaki Y. Disruption of cytoskeletal structures mediates shear stress-induced endothelin-1 gene expression in cultured porcine aortic endothelial cells. J.
Clin. Invest. 92: 1706-1712, 1993.
12. Morita T, Yoshizumi M, Kurihara H, Maemura K,
Nagai R, Yazaki Y. Shear stress increases heparinbinding epidermal growth factor-like growth factor
mRNA levels in human vascular endothelial cells.
Biocem. Biophys. Res. Commun. 197: 256-262, 1993.
13. Kurihara Y*, Kurihara H*, Suzuki H, Kodama T,
Maemura K, Nagai R, Oda H, Kuwaki T, Cao W-H,
Kamada N, Jishage K, Ouchi Y, Azuma S, Toyoda Y,
Ishikawa T, Kumada M, Yazaki Y. Elevated bloodpressure and craniofacial abnormalities in mice deficient in endothelin-1. Nature 368: 703-710, 1994.
(*equal contribution)
14. Maemura K, Kurihara H, Kurihara Y, Nagai R, Yazaki
Y. Isolation and characterization of vascular endothelial cells derived from mice lacking endothelin-1.
Biochem. Biophys. Res. Commun. 201: 538-545,
1994.
15. Maemura K, Kurihara Y, Morita H, Kurihara H,
Yazaki Y. Renal endothelin and hypertension - reply.
Nature 372: 50, 1994.
16. Morita T, Kurihara H, Maemura K, Yoshizumi M,
Nagai R, Yazaki Y. Role of Ca2+ and protein kinase C
in shear stress-induced actin depolymerization andendothelin-1 gene expression. Circ. Res. 75: 630-636,
1994.
17. Harats D*, Kurihara H*, Belloni P, Oakley H, Ziober
A, Ackley D, Cain G, Kurihara Y, Lawn R, Sigal E.
Targeting gene expression to the vascular wall intransgenic mice using the murine preproendothelin-1
promoter. J. Clin. Invest. 95: 1335-1344, 1995.
(*equal contribution)
18. Kurihara Y, Kurihara H, Oda H, Maemura K, Nagai R,
Ishikawa T, Yazaki Y. Aortic arch malformations andventricular septal defect in mice deficient in endothelin-1. J. Clin. Invest. 96: 293-300, 1995.
19. Maemura K, Kurihara H, Kurihara Y, Oda H,
Ishikawa T, Copeland NG, Gilbert DJ, Jenkins NA,
Yazaki Y. Sequence analysis, chromosomal locationand developmental expression of the mouse preproendothelin-1 gene. Genomics 31: 177-184, 1996.

28

20. Kuwaki T, Cao W-H, Kurihara Y, Kurihara H, Ling GY, Onodera M, Ju K-H, Yazaki Y, Kumada M.
Impaired ventilatory responses to hypoxia andhypercapnia in mutant mice deficient in endothelin1. Am. J. Physiol. 270: R1279-R1286, 1996.
21. Reid K, Turnley AM, Maxwell GD, Kurihara Y,
Kurihara H, Bartlett PF, Murphy M. Multiple roles forendothelin 3 in melanocyte development: Regulationof progenitor number and stimulation of differentiation. Development 122: 3911-3919, 1996.
22. Minamino T, Kurihara H, Takahashi M, Shimada K.
Maemura K, Oda H, Ishikawa T, Uchiyama T,
Tanzawa K, Yazaki Y. Endothelin converting enzyme(ECE-1) expression in the rat vascular injury modeland human coronary atherosclerosis. Circulation 95:
221-230, 1997.
23. Suzuki H, Kurihara Y, Takeya M, Kamada N, Kataoka
M, Jishage K, Ueda O, Sakaguchi H, Higashi T,
Suzuki T, Takashima Y, Kawabe Y, Cynshi O, Wada
Y, Honda M, Kurihara H, Aburatani H, Doi T,
Matsumoto A, Azuma S, Noda T, Toyoda Y, Itakura
H, Yazaki Y, Horiuchi S, Takahashi K, Kruijt JK, vanBerkel TJC, Steinbrecher UP, Ishibashi S, Maeda N,
Gordon S, Kodama T. A role for macrophage scavenger receptors in atherosclerosis and susceptibility
to infection. Nature 386: 292-296, 1997.
24. Kurihara H, Kurihara Y, Maemura K, Yazaki Y. The
role of endothelin-1 in cardiovascular development.
Ann. NY. Acad. Sci. 811: 168-177, 1997.
25. Morita H, Taguchi J, Kurihara H, Kitaoka M, Kaneda
H, Kurihara Y, Maemura K, Shindo T, Minamino T,
Ohno M, Yamaoki K, Ogasawara K, Aizawa T, Suzuki
S, Yazaki Y. Genetic polymorphism of 5,10-methylenetetrahydrofolate reductase (MTHFR) as a risk factor for coronary artery disease. Circulation 95: 20322036, 1997.
26. Kuwaki T, Kurihara H, Cao WH, Kurihara Y,
Unekawa M, Yazaki Y, Kumada M: Physiological roleof brain endothelin in the central autonomic control:from neuron to knockout mice. Prog. Neurobiol. 51:
545-579, 1997.
27. Nagase T, Kurihara H, Kurihara Y, Aoki-Nagase T,
Fukuchi Y, Yazaki Y, Ouchi Y. Airway hyperresponsiveness to methacholine in mutant mice deficient inendothelin-1. Am. J. Respir. Crit. Care Med. 157: 560564, 1998.
28. Kitano Y, Kurihara H, Kurihara Y, Maemura K, Ryo
Y, Yazaki Y, Harii K. Gene expression of bone matrix
proteins and endothelin receptors in endothelin-1deficient mice revealed by in situ hybridization. J.
Bone Miner. Res. 13: 237-244, 1998.
29. Thomas T, Kurihara H, Yamagishi H, Kurihara Y,
Yazaki Y, Olson EN, Srivastava D. A signaling cascade involving endothelin-1, dHAND and Msx1 regulates development of neural crest-derived branchialarch mesenchyme. Development 125: 3005-3014,
1998.
30. Morita H, Kurihara H, Tsubaki S, Sugiyama T,
Hamada C, Kurihara Y, Shindo T, Oh-hashi Y,
Kitamura K, Yazaki Y. Methylenetetrahydrofolatereductase (MTHFR) gene polymorphism andischemic stroke in Japanese. Arterioscler. Thromb.
Vasc. Biol. 18: 1465-1469, 1998.
31. Ling GY, Cao WH, Onodera M, Ju KH, Kurihara H,
Kurihara Y, Yazaki Y, Kumada M, Fukuda Y, Kuwaki
T. Renal sympathetic nerve activity in mice: comparison between mice and rats and between normal andendothelin-1 deficient mice. Brain Res. 808: 238-249,
1998.
32. Ohashi Y, Kawashima S, Hirata K, Yamashita T,
Ishida T, Inoue N, Sakoda T, Kurihara H, Yazaki Y,
Yokoyama M. Hypotension and reduced nitric oxideelicited vasorelaxation in transgenic mice overexpressing endothelial nitric oxide synthase. J. Clin.
Invest. 102: 2061-2071, 1998.
33. Morita H, Kurihara H, Sugiyama T, Kurihara Y,
Shindo T, Oh-hashi Y, Yazaki Y. D919G polymorphism of the methionine synthase gene. Association
with homocysteine metabolism and late-onset vascular diseases in the Japanese population. Arterioscler.
Thromb. Vasc. Biol. 19: 298-302, 1999.
34. Kuwaki T, Ling GY, Onodera M, Ishii T, Nakamura
A, Ju KH, Cao WH, Kumada M, Kurihara H, Kurihara
Y, Yazaki Y, Ohuchi T, Yanagisawa M, Fukuda Y.
Endothelin in the central control of cardiovascularand respiratory functions. Clin. Exp. Pharmacol.
Physiol. 26: 989-994, 1999.
35. Kurihara H, Kurihara Y, Nagai R, Yazaki Y.
Endothelin and neural crest development. Cell. Mol.
Biol. 45: 639-651, 1999.
36. Nagase T, Kurihara H, Kurihara Y, Aoki-Nagase T,
Nagai R, Ouchi Y. Disruption of ET-1 gene enhances
pulmonary responses to methacholine via functional
mechanism in knockout mice. J. Appl. Physiol. 87:
2020-2024, 1999.
37. Shindo T, Kurihara H, Maemura K, Kurihara Y,
Kuwaki T, Izumida T, Minamino N, Ju K-H, Morita
H, Oh-hashi Y, Kumada M, Kangawa K, Nagai R,
Yazaki Y. Hypotension and resistance to lipopolysaccharide-induced shock in transgenic mice overexpressing adrenomedullin in their vasculature.
Circulation 101: 2309-2316, 2000.
38. Kurihara Y*, Kurihara H*, Morita H, Cao W-H, Ling
G-Y, Kumada M, Kimura S, Nagai R, Yazaki Y,
Kuwaki T. Role of endothelin-1 in stress response in
the central nervous system. Am. J. Physiol. 279:
R515-R521, 2000. (*equal contribution)
39 Shindo T, Kurihara H, Kuno K, Yokoyama H, Wada
T, Kurihara Y, Imai T, Wang Y, Ogata M, Oh-hashi Y,
Morita H, Ishikawa T, Nagai R, Yazaki Y,
Matsushima K. ADAMTS-1: a metalloproteinase-disintegrin essential for normal growth, fertility andorgan morphology and function. J. Clin. Invest. 105:1345-1352, 2000.
40 Harats D, Shaish A, George J, Mulkins M, Kurihara
H, Levkovitz H, Sigal E. Overexpression of 15-lipoxygenase in vascular endothelium accelerates early atherosclerosis in LDL receptor-deficient mice.
Arterioscler. Thromb. Vasc. Biol. 20: 2100-2105,
2000.

41 Morita H, Kurihara H, Yoshida S, Saito Y, Shindo T,
Oh-hashi Y, Kurihara Y, Yazaki Y, Nagai R. Dietinduced hyperhomocysteinemia exacerbates neointima formation in rat carotid arteries after baloon
injury. Circulation 103: 133-139, 2001.
42 Ho MCY, Lo ACY, Kurihara H, Yu ACH, Chung SK.
Endothelin-1
protects
astrocytesfromhypoxic/ischemic injury. FASEB J. 15: 618-626, 2001.
43 Imai T, Morita T, Shindo S, Nagai R, Yazaki Y,
Kurihara H, Suematsu M, Katayama S. Vascularsmooth muscle cell-directed overexpression ofhuman heme oxygenase-1 elavates blood pressure
through attenuation of NO-induced vasodilation in
mice. Circ. Res. 89:55-62, 2001.
44 Yamaguchi Y, Fukuhara S, Nagase T, Tomita T,
Hitomi S, Kimura S, Kurihara H, Ouchi Y. A novel
mouse ß-defensin, mBD-6, predominantly expressedin skeletal muscle. J. Biol.Chem. 276:31510-31514,
2001.
45 Imai Y, Shindo T, Maemura K, Kurihara Y, Nagai R,
Kurihara H. Evidence for the physiological andpathological roles of adrenomedullin from genetic
engineering in mice. Ann. NY. Acad. Sci. 947:26-34,
2001.
46 Shindo T, Kurihara Y, Nishimatsu H, Moriyama N,
Kakoki M, Wang Y, Imai Y, Ebihara A, Kuwaki T, Ju
K-H, Minamino N, Kangawa K, Ishikawa T, Fukuda
M, Akimoto Y, Kawakami H, Imai Y, Imai T, Morita
H, Yazaki Y, Nagai R, Hirata Y, Kurihara H. Vascularabnormalities and elevated blood pressure in micelacking adrenomedullin gene. Circulation 104:19641971, 2001.
47 Oh-hashi Y, Shindo T, Kurihara Y, Imai T, Wang Y,
Morita H, Imai Y, Kayaba Y, Nishimatsu H, Suematsu
Y, Hirata Y, Yazaki Y, Nagai R, Kuwaki T, Kurihara
H. Elevated sympathetic nervous activity in mice
deficient in aCGRP. Circ. Res. 89:983-990, 2001.
48 Shindo T, Kurihara H, Maemura K, Kurihara Y, Ueda
O, Suzuki H, Kuwaki T, Ju K-H, Wang Y, Morita H,
Oh-hashi Y, Kumada M, Nagai R, Yazaki, Kimura K.
Renal damage and salt-dependent hypertension inaged transgenic mice overexpressing endothelin-1. J.
Mol. Med. 80:105-116, 2002.
49 Imai Y, Shindo T, Sata M, Memura K, Saito Y,
Akishita M, Osuga J, Ishibashi S, Tobe K, Maekawa
H, Oh-hashi Y, Morita H, Kurihara Y, Yazaki Y, Nagai
R, Kurihara H. Resistance to neointimal hyperplasia
and fatty streak formation in adrenomedullin overexpression mice. Arterioscler. Thromb. Vasc. Biol.
22: 1310-1315, 2002.
50. Yamaguchi Y, Nagase T, Makita R, Fukuhara S,
Tomita T, Tominaga T, Kurihara H, Ouchi Y.
Identification of multiple novel epididymis-specificb-defensin isoforms in humans and mice. J.
Immunol. 169: 2516-2523, 2002.

29

Department ofIntegrative Physiology
Outline and Research Objectives
This department was initially established in 1877 as the First Department of Physiology,
and reorganized in 1997 as the Department of Integrative Physiology. Our department collaborates with other laboratories dedicated to Physiological Sciences, that is, theDepartment of Molecular/Cellular Physiology and the Department of Neurophysiology, inteaching activities for undergraduate courses and the nursing school. The fields in which
our department specializes include the entire spectrum of the physiology of “animal functions”, such as general physiology, sensory physiology, neurophysiology, higher nervous
functions and cognitive neurosciences.

Faculties and Students
Professor and Chair Yasushi Miyashita, Ph.D. (1989- )Lecturer
Isao Hasegawa, M.D., Ph.D. (2000- )Associate .................................2
Postdoctoral Fellow ................5
Graduate Student..................10
Research Student ....................1
Secretary .................................3

lus-selectivity of IT neurons can be acquired throughlearning in adulthood. Second, the activity of IT neurons can link the representations of temporally associated but geometrically unrelated stimuli.

Most of our research has been focused on thehigher brain functions of the mammalian central nervous system, in particular, the neural mechanisms of
cognitive memory in the primate. The basic motivation of the research can be best explained as follows:
Knowledge or experiences are voluntarily recalledfrom memory by reactivation of their neural representations in the cerebral association cortex. Three questions are central in the understanding of this process:(1) Where are mnemonic representations coded andhow are they organized?
(2) Which neural processes create the representation?
(3) What is the mechanism underlying reactivation of
the representation on demand of voluntary recall?

2. Activating the representation in the temporal
cortex on demand
In spite of the classical clinical observation thatelectric stimulation of the temporal lobe produces‘experiential responses’, there have been no direct evidence supporting the notion of ‘reactivation of neuralrepresentations’ during memory retrieval. We firstreported a neuronal correlate of the reactivation
process as ‘pair-recall neurons’ [42, 45]. In our subsequent study [30], we devised a new modified pairassociate task in which the necessity for memory
retrieval and its initiation time were controlled by acolour switch, independent of the cue stimulus presentation. Single-unit recordings in monkeys performing this task revealed that IT neurons selective to amemorized object are dynamically activated at thetime of memory retrieval of that object, and suppressed at the time of retrieval of other objects. Then,it became important to determine the neural networkthat drives the memory retrieval machinery in the IT
cortex.

1. Creating the mnemonic representation
Lesion studies in primates have implicated the IT
cortex in long-term memory storage of visual objects[40]. Neuronal correlates of associative long-term
memory were first discovered in the IT cortex byMiyashita [49, 50] and Sakai & Miyashita [45]. Hedeveloped a novel memory paradigm that requires the
subject to create a link of associative memory
between mathematically designed pictures (for examples, see Fig.1). Their single-unit recording experiments in thepair-association task identified two
mnemonic properties of IT neurons. First, the stimu-

Figure 1 : Paired associates. Each pictures was created according to a fractal algorithm.

Past Research and Major Accomplishments30

Figure 2 : Which information is carried via top-down signals? (a) A stimulus-stimulus association task. Twenty cue-pictures are randomly sorted into five categories. Each of the four cues in one category specifies a common choice. (b) Category-selective delay
activity of an inferior temporal neuron. Delay activities were raised for all cues in Category I, but not for any cues in Category V(rastergrams, top-down condition). Choice responses are also strongest for Category I and weakest for Category V. Spike density
functions show averaged activities across four cues in Category I (thick) and in Category V (thin) for both conditions (top-down,
blue; bottom-up, black). (c) Spike density functions of the top-down response for five categories, as well as the choice responses,
shown by a pseudo-colour coding. Note that the category-selective delay activity can predict choice selection.

3. Top-down activation through fronto-temporalpathway
A candidate component of the neural circuit is the
top-down activation from the prefrontal cortex. The
prefrontal cortex has been implicated in various executive processes, and its contribution to mnemonic
functions, particularly in episodic memory and working memory, is repeatedly demonstrated in humanneuroimaging studies. We attempted to directly test
its contribution to memory retrieval control by thecapacity for interhemispheric transfer of mnemonic
signal through the anterior corpus callosum, a key
structure interconnecting prefrontal cortices. We
introduced the posterior-split-brain paradigm into theassociative memory task in monkeys [23]. Long-term
memory acquired through stimulus-stimulus association did not interhemispherically transfer via theanterior corpus callosum. Nonetheless, when a visualcue was presented to one hemisphere, the anteriorcallosum could instruct the other hemisphere to
retrieve the correct stimulus specified by thecue. Therefore, although visual long-term memory is
stored in the temporal cortex, memory retrieval isunder the executive control of the prefrontal cortex.
In spite of predictions based on these behaviouralexperiments, no neuronal correlate of the top-down
signal from the prefrontal cortex to IT cortex had
been detected. We provided the first evidence of theexistence of the top-down signal [14] by conducting
single-unit recording in posterior-split-brainmonkeys. In the absence of bottom-up visual inputs,
single IT neurons were robustly activated by the topdown signal, which conveyed information on semantic categorization imposed by visual stimulus-stimulus
association (Fig. 2). We also demonstrated that the

top-down signal had a longer latency by about 100 ms
than the bottom-up signal. The longer latency is most
likely ascribed to a multi-synaptic conduction delaythat reflected the signal transformation within theprefrontal cortices.

Current Research and Future Prospects
Most of our current research is focused on the third
question on the mechanisms of cognitive memory, thatis, (3) What is the mechanism underlying reactivationof the representation on demand of voluntary recall?
Two complementary approaches are of interest in
answering the above question. First, neuroimaging
studies in humans can elucidate brain representations
and their interactions in high-level memory processes,
which have been conceptualized as retrieval attempt
or contextual monitoring. Second, the well-analyzed
neural representations in the monkey cortex should
be compared with those in the human cortex. The
comparison would provide strong evidence that couples active neural codes in the brain network withconscious experiences.
An example of our recent achievements along thefirst approach is the discovery of a neural correlate of
the “feeling-of-knowing” [1]. The “feeling-of-knowing(FOK)” is a subjective sense of knowing a word before
recalling it, and it provides us clues to understanding
the mechanisms of human meta-memory systems.
We investigated neural correlates for the FOK basedon the recall-judgment-recognition paradigm. Eventrelated functional magnetic resonance imaging (fMRI)
with parametric analysis was used. We found activations in the left dorsolateral, left anterior, bilateralinferior, and medial prefrontal cortices that signifi-

31

Figure 3 : Comparison of activated cortical areas in set-shifting of WCST in monkeys (left) and humans (right).

cantly increased as the FOK became greater.
Furthermore, we demonstrated that the FOK-region
in the right inferior frontal gyrus and a subset of the
FOK-region in the left inferior frontal gyrus are notrecruited for successful recall processes, suggesting
their particular role in meta-memory processing.
Along the second approach, we have recently
demonstrated that fMRI studies in macaque monkeys
allow us to directly compare the brain activity ofhumans with that of monkeys with respect to highlevel cognitive functions [2]. In this study, the functional brain organization of macaque monkeys andhumans was directly compared by fMRI. Subjects ofboth species performed a modified Wisconsin Card
Sorting Test that required behavioural flexibility inthe form of cognitive set shifting. Equivalent visual
stimuli and task sequence were used for the two
species. We found transient activation related to cognitive set shifting in focal regions of the prefrontal
cortex in both monkeys and humans. These functional homologs were located in cytoarchitectonicallyequivalent regions in the posterior part of the ventrolateral prefrontal cortex. This comparative imaging
provides insights into the evolution of cognition inprimates.
Various neuroimaging studies have been carriedout to clarify cognitive functions of the humanbrain. Because most neuroimaging studies rely on thecorrelation between cognitive processes and brain
activations, conjunction with other complementary
methods, such as neuropsychological studies andtranscranial magnetic stimulation, should be promoted to clarify the behavioural significance of observed
brain activities. Emerging fMRI of the monkey brain
enables us to directly compare the brain activity ofhumans with that of monkeys using the same modality. Therefore the monkey fMRI will make it possible
to combine imaging studies with electrophysiology orlesion studies, and will hopefully lead to the understanding of the causal relationship between activatedbrain areas and cognitive functions. Since most of thedetailed knowledge of anatomy and function of thecerebral cortex has come from studies in monkeys,
sharing the same method among the studies of monkeys and humans would advance the understandingof the neural basis of human cognition.

32

Research Grants
1. A Grant-in-Ai32d for Specially Promoted Research,
MEXT (07102006)
Neural mechanisms of cognitive memory system :Integration of functional magnetic resonance imaging method and molecular/cellular approaches.
¥ 215,000,000 (1995-1999)
2. A Grant-in-Aid for Specially Promoted Research,
MEXT (Extension 07102006)
Neural mechanisms of cognitive memory system :Integration of functional magnetic resonance imaging method and molecular/cellular approaches.
¥309,000,000 (2000-2001)
3. A Grant-in-Aid for Specially Promoted Research,
MEXT (14002005)
Brain distributed network : Integrative study basedon functional MRI in monkeys¥ 550,000,000 (2002-2006)

Select Publications
1. Kikyo, H., Ohki, K. and Miyashita, Y. Neural correlates for “feeling-of-knowing”: an fMRI parametricanalysis.
Neuron 36, 177-186, 2002.
2. Nakahara, K., Hayashi, T.,Konishi, S. and Miyashita,
Y. Functional MRI of macaque monkeys performinga cognitive set-shifting task.
Science 295, 1532-1536, 2002.
3. Konishi, S., Hayashi, T., Uchida, I., Kikyo, H.
Takahashi, E. and Miyashita, Y. Hemispheric asymmetry in human lateral frontal cortex during congnitive set shifting.
Proc. Natl. Acad. Sci. USA 99, 7803-7808, 2002.
4. Hasegawa, I. and Miyashita, Y. Making categories:
expert neurons look into key features.
Nature Neurosci. 5, 90-91, 2002.
5. Tokuyama, W., Okuno, H., Hashimoto, T., Li, X.Y.
and Miyashita, Y. Selective zif268 mRNA inductionin the perirhinal cortex macaque of monkeys duringformation of visual pair-association memory.
J. Neurochem. 81, 60-70, 2002.
6. Matsuzaki, M., Ellis-Davies, G.C.R., Nemoto, T.,
Miyashita, Y., Iino, M. and Kasai, H. Dendritic spine
geometry is critical for AMPA receptors expression
in hippocampal CA1 Pyramidal neurons.
Nature Neurosci. 4, 1086-1092, 2001.

7. Naya, Y., Yoshida, M. & Miyashita, Y. Backward
spreading of memory retrieval signal in the primate
temporal cortex.
Science 291, 661-664, 2001.
8. Li, X.Y., Hashimoto, T., Tokuyama, W., Miyashita,
Y. and Okuno H Spatio-temporal dynamics of BDNF
mRNA induction in the vestibulo-olivary network
during vestibular compensation.
J. Neurosci 21, 2738-2748, 2001.
9. Miyashita,Y. and Farah, M.J. Cognitive neuroscienceat the turn of the millennium.
Curr. Opin. Neurobiol. 11, 147-149, 2001.
10. Nemoto, T., Kimura, R., Ito,K., Tachikawa, A.,
Miyashita, Y., Iino, M. and Kasai, H. Sequential
replenishment mechanism of exocytosis in pancreatic acini.
Nature Cell Biol. 3, 253-258, 2001.
11. Tokuyama,W., Okuno,H., Hashimoto, T., Li, Y.X.
and Miyashita, Y. BDNF upregulation during declarative memory formation in monkey inferior temporal
cortex.
Nature Neurosci.3, 1134-1142, 2000.
12. Embick, D., Marantz, A., Miyashita, Y., O’Neil W.
and Sakai, K.L. A syntactic specialization for Broca’s
area.
Proc. Natl. Acad. Sci. USA 97, 6150-6154, 2000.
13. Miyashita, Y. and Hayashi, T. Neural representationof visual objects : encoding and top-down activation.
Curr. Opin. Neurobiol. 10, 187-194, 2000.
14. Tomita, H., Ohbayashi, M., Nakahara, K., Hasegawa,I. and Miyashita, Y. Top-down signal originatingfrom the prefrontal cortex for memory retrieval.
Nature 401, 699-703, 1999.
15. Maeda, H., Ellis-Davies, G.C.R., Ito, K., Miyashita, Y.
and Kasai, H. Supralinear Ca2+ signaling by cooperative and mobile Ca2+ buffering in Purkinje neurons.
Neuron 24, 989-1002, 1999.
16. Kasai, H., Kishimoto, T., Liu, T., Miyashita, Y.,
Podini, P., Grohovaz, F. and Meldolesi, J. Multiple
and diverse forms of regulated exocytosis in wildtype and defective PC12 cells.
Proc. Natl. Acad. Sci. USA 96, 945-949, 1999.
17. Konishi, S., Nakajima, K., Uchida, I., Kikyo, H.,
Kameyama, M. and Miyashita, Y. Common inhibitory mechanism in human inferior prefrontal cortex
revealed by event-related functional MRI.
Brain 122, 981-991, 1999.
18. Konishi, S., Kawazu, M., Uchida, I., Kikyo. H.,
Asakura, I. and Miyashita, Y. Contribution of working memory to transient activation in human inferior
prefrontal cortex during performance of Wisconsin
card sorting test.
Cerebral Cortex 9, 745-753, 1999.
19. Okuno, H., Tokuyama, W., Li, Y-X., Hashimoto, T.
and Miyashita, Y. Quantitative evaluation of neurotrophin and trk mRNA expression in visual andlimbic areas along the occipito-temporo-hippocampal
pathway in adult macaque monkeys.
J. Comp. Neurol. 408, 378-398, 1999.

20. Uchida, I., Kikyo, H., Nakajima, K., Sekihara, K. andMiyashita, Y. Activation of lateral extrastriate areas
during orthographic processing of Japanese characters studied with fMRI.
Neuroimage 9, 208-215, 1999.
21. Ito, K., Miyashita, Y. and Kasai H Kinetic control ofmultiple forms of Ca2+ spikes by inositol trisphosphate in pancreatic acinar cells.
J. Cell Biol. 146, 405-413, 1999.
22. Takahashi, N., Kadowaki, T., Yazaki, Y., Ellis-Davies,
G.C.R., Miyashita, Y. and Kasai, H. Post-priming
actions of ATP on Ca2+-dependent exocytosis in pancreatic beta cells.
Proc. Natl. Acad. Sci. USA 96, 760-765, 1999.
23. Hasegawa, I., Fukushima, T., Ihara, T. andMiyashita, Y. Callosal window between prefrontal
cortices : cognitive interaction to retrieve long-term
memory.
Science 281, 814-818, 1998.
24. Konishi, S., Nakajima, K., Uchida, I., Kameyama, M.,
Nakahara, K., Sekihara K. and Miyashita, Y.
Transient activation of inferior prefrontal cortex during cognitive set shifting.
Nature Neurosci.1, 80-84, 1998.
25. Konishi, S., Nakajima, K., Uchida, I., Sekihara, K. andMiyashita, Y. Temporally resolved no-go dominant
brain activity in the prefrontal cortex revealed byfubctional magnetic resonance imaging.
Neuroimage 5, 120, 1997.
26. Takahashi, N., Kadowaki, T., Yazaki, Y., Miyashita,
Y. and Kasai, H. Multiple exocytotic pathways inpancreatic β cells.
J. Cell Biol. 138, 55-64, 1997.
27. Ninomiya, Y., Kishimito, T., Yamazawa, T., Ikeda,
H., Miyashita, Y. and Kasai, H. Kinetic diversity inthe fusion of exocytotic vesicles.
EMBO J. 16, 929-934, 1997.
28. Ito, K., Miyashita, Y. and Kasai H. Micromolar andsubmicromolar Ca2+ spikes regulating distinct cellularfunctions in pancreatic acinar cells.
EMBO J. 16, 242-251, 1997.
29. Higuchi, S. and Miyashita, Y. Formation of mnemonic neuronal responses to visual paired associates ininferotemporal cortex is impaired by perirhinal andentorhinal lesions.
Proc. Natl. Acad. Sci. USA, 93, 739-743, 1996.
30. Naya, Y., Sakai, K. and Miyashita, Y. Activity of primate inferotemporal neurons related to a sought target in pair-association task.
Proc. Natl. Acad. Sci. USA, 93, 2664-2669, 1996.
31. Kasai, H., Takagi, H., Ninomiya, Y., Kishimoto, T.,
Ito, K., Yoshida, A., Yoshioka., T. and Miyashita, Y.
Two components of exocytosis and endocytosis inpheochromocytoma cells studied using caged-Ca2+
compounds.
J. Physiol. 494, 53-65, 1996.
32. Ninomiya, Y., Kishimoto, T., Miyashita, Y. andKasai, H. Ca2+-dependent exocytotic pathways in chinese hamster ovary fibroblasts revealed by a cagedCa2+ compound.

33

J. Biol. Chem. 271, 17751-17754, 1996.
33. Sakai, K., Watanabe, E., Onodera, Y., Uchida, I.,
Kato, H., Yamamoto, E., Koizumi, H. and Miyashita,
Y. Functional mapping of the human colour centre
with echo-planar magnetic resonance imaging.
Proc. R. Soc. Lond. B 261, 89-98, 1995.
34. Miyashita, Y. : How the brain creates imagery:
Projection to primary visual cortex.
Science 268, 1719-1720, 1995.
35. Okuno, H., Saffen, D.W. and Miyashita, Y.
Subdivision-specific expression of Zif268 in the hippocampal formation of the Macaque monkey.
Neuroscience 66, 829-845, 1995.
36. Sakai, K., Watanabe, E., Onodera, Y., Itagaki, H.,
Yamamoto, E., Koizumi, H. and Miyashita, Y.
Functional mapping of the human somatosensory
cortex with echo-planar magnetic resonance imaging.
Magn. Reson. Med. 33, 736-743, 1995.
37. Sakai, K. and Miyashita, Y. Visual imagery : An interaction between memory retrieval and focal attention.
Trends Neurosci. 17, 287-289, 1994.
38. Mori, A., Takahashi, T., Miyashita, Y. and Kasai, H.
Two distinct glutamatergic synaptic inputs to thestriatal medium spiny neurones of neonatal rat andpaired-pulse depression.
J. Physiol. 476, 217-228, 1994.
39. Kasai, H., Li, Yeu Xin and Miyashita, Y. Subcellular
distribution of Ca2+ release channels underlying Ca2+
waves and oscillations in exocrine pancreas.
Cell 74, 669-677, 1993.
40. Miyashita, Y. Inferior temporal cortex: where visual
perception meets memory.
Annu. Rev. Neurosci. 16, 245-263, 1993.
41. Sakai, K. and Miyashita, Y. Memory and imagery in
the Temporal lobe.
Curr. Opin. Neurobiol. 3, 166-170, 1993
42. Miyashita, Y., Date, A. and Okuno, H.
Configurational encloding of complex visual forms
by single neurons of monkey temporal cortex.
Neuropsychologia 31, 1019-1031, 1993.
43. Maruyama, Y., Inooka, G., Li, Y., Miyashita, Y. andKasai, H. Agonist-induced locarized Ca2+ rises directly
triggering exocytotic secretion in exocrine pancreas.
EMBO J. 12, 3017-3022, 1993.
44. Iino, M., Yamazawa, T., Miyashita, Y., Endo, M. andKasai, H. Critical intracellular Ca2+ concentration for
all-or-none Ca 2+ spiking in single smooth muscle
cells.
EMBO J. 12, 5287-5291, 1993.
45. Sakai, K. and Miyashita, Y. Neural organization forthe long-term memory of paired associates.
Nature 354, 152-155, 1991.
46. Rolls, E.T., Miyashita, Y., Cahusac, P., Kesner, R.P.,
Niki, H., Feigenbaum, J. and Bach, L. Hippocampal
neurons in the monkey with activity related to theplace in which a stimulus is shown.
J. Neurosci. 9, 1835-1845, 1989.
47. Mori, K. and Miyashita, Y. Localized metabolic
responses to optokinetic stimulation in the brain

34

stem nuclei and the cerebellum investigated with the[14C]2-deoxyglucose method in rats.
Neuroscience 30, 271-281 1989.
48. Miyashita, Y., Rolls, E.T., Cahusac, P. and Niki, H.
Activity of hippocampal formation neurons in themonkey related to a stimulus-response association
task.
J. Neurophysiol. 61, 669-678, 1989.
49. Miyashita, Y. Neuronal correlate of visual associative
long-term memory in the primate temporal cortex.
Nature 335, 817-820, 1988.
50. Miyashita, Y. and Chang, H.S. Neuronal correlate of
pictorial short-term memory in the primate temporal
cortex.
Nature 331, 68-70, 1988.

Department of Cellular andMolecular Physiology
Outline and Research Objectives
Knowledge comes to man through the door of the senses (Heraclitus). Using multidisciplinary approaches including electrophysiology, optical imaging, molecular / cell biology, andmolecular genetics, we aim at better understanding of neuronal mechanisms for the sensory
perception of the external world and for the emotional state induced in the brain by thesensory inputs. For this purpose we are currently analyzing the central nervous system forolfaction, a sensory modality that has a strong influence to human emotion. Another major
research focus of this department is to understand cellular and molecular mechanisms forcontact-mediated interactions between neurons and immune cells that occur in pathologicaland physiological conditions.

Faculties and Students
Professor and Chair Kensaku Mori, Ph. D. (May 1999 ~)Lecturer
Masahiro Yamaguchi, M.D., Ph. D.
Associate .................................2
Graduate student ..................12
Research student.....................1
Secretary .................................1(e.g., telencephalin (TLCN), Yoshihara et al., Neuron,
1994; OCAM, Yoshihara et al., J. Neurosci., 1997). We
are currently extending the analysis of the functionalroles of these cell adhesion molecules using TLCNdeficient mice (in collaboration with Prof. Mishina)
and OCAM-deficient mice. During the course of theanalysis of TLCN, we unexpectedly found that thebinding between TLCN and its counterreceptor LFA-1
mediated the interaction between neurons andPast Research and Major Accomplishments
K. Mori and colleagues have been studying thefunctional organization of the olfactory nervous system for nearly 28 years. Major accomplishment of ourresearch group before 1999 includes (1) a systematicfunctional and structural analysis of the neuronal circuits in the mammalian olfactory bulb, the first centerfor the processing of olfactory information in thebrain, (2) the finding that individual principal neurons
in the olfactory bulb respond selectively to a range ofodorants having common molecular structures, and(3) the finding of zone-to-zone axonal connectivity
pattern between olfactory sensory epithelium and theolfactory bulb. After K. Mori’s arrival at the
Department of Cellular and Molecular Physiology in
May 1999, we further extended our previous studies
and found a clue for understanding the spatial organization of the ‘odor map’ in the olfactory bulb (Fig.1; e.
g., Mori et al., Science, 1999, Uchida et al., Nature
Neurosci., 2000., Nagao et al., NeuroReport, 2000).
These studies as well as our novel molecular genetic
approaches (Yoshihara et al., Neuron 1999) formed abasis for current and future studies on the ‘odor maps’
in the olfactory bulb and olfactory cortex (e.g., Inaki
et al., Eur. J. of Neurosci., 2002).
We previously found several novel neuronal celladhesion molecules and analyzed their functionalroles in the formation of neuronal circuit in the brain

35

immune cells (Mizuno et al., J. B. C., 1997; Tian et al.,J. Cell Biol., 2000, Eur. J. Immunol., 2000, J. Immunol.,1997). We thus developed a co-culture assay system to
examine the contact-mediated interactions between
neurons and immune cells.
Our group generated transgenic mice in which
newly generated neurons are visualized in the brain
by the green fluorescent protein (Yamaguchi et al.,NeuroReport, 2000). Using the transgenic mice, we are
now studying the neurogenesis and neuron-elimination in the adult brain.

Current Research
Current research programs and targets can be categorized into the following four topics.
(1) Functional analysis of the neuronal circuits inthe central olfactory nervous system
An astonishing feature of the olfactory system is
its ability to distinguish among more than 400,000
different odorants and among countless number ofodorants-mixtures, each having specific ‘odor’. Using
optical imaging and electrophysiological methods, weare studying the ‘odor maps’ in the central olfactory
system to understand the logic employed by the olfactory system for discrimination among numerous odorants and for perception and learning of the olfactory
image of objects. The target regions for the mapping
include the olfactory bulb and several regions of theolfactory cortex (e.g., piriform cortex and olfactory
tubercle). In addition, odorants having pleasant orunpleasant ‘odor’ are used to map the brain regions
and to elucidate neuronal mechanisms responsible forthe emotional states of the brain.
(2) Neurogenesis and neuron-elimination in theadult brain
Olfactory nervous system has an unusual capacity
to generate neurons throughout life. The sensory neurons in the nasal epithelium and the local interneurons in the olfactory bulb are turning over continuously even in the adult. Therefore, we choose the olfactory system as a model system with which to study themolecular and cellular mechanisms for the recruitment of newly generated neurons into the adult central nervous system and for the selective eliminationof damaged neurons from the functional neuronal circuit. Molecular-genetic and physiological methods are
combined to analyze the mechanisms of the neuronrecruitment and neuron-elimination. The knowledgeof the new-neuron recruitment into the functioning
neuronal circuit should be of critical importance in theremedy of neurological diseases that accompany neuron loss.

36(3) Molecular and cellular mechanisms for theaxonal recognition of specific target neurons andfor the formation of specific neuronal circuits
During development, growing axons, including theaxons from olfactory sensory neurons, can find their
specific target neurons and form functional neuronal
circuits. Recent studies on olfactory sensory neurons
demonstrate a remarkable relationship between theselection of a single odorant receptor gene among arepertoire of 1000 and the address of the axon-projection target-glomeruli in the olfactory bulb. Suchknowledge on the detailed address of the axon targets
is currently available only in the olfactory system. We
apply proteomics approaches to the olfactory axon
address system to elucidate molecular mechanisms
for the olfactory axon guidance to the target glomeruli
and for the formation of specific synaptic connections
with target neurons in the olfactory bulb.
(4) Cellular and molecular mechanisms for thecontact-mediated interactions between neurons
and immune cells in physiological and pathological conditions
Immune cells rarely meet central neurons in physiological conditions because of the blood brain barrier.
However, a massive infiltration of immune cells into
the brain occurs under many pathological conditions,suggesting that immune cells may directly interact
with central neurons. To understand the nature andconsequence of the direct interaction between
immune cells and neurons, we are examining thechange in neuronal morphology and physiologicalstate of primary cultured neurons following the coculture with immune cells. In addition, we are currently focusing on telencephalon-neuron specificmembrane protein, telencephalin (TLCN), which bindto LFA-1 integrin expressed by leukocytes. Since thesoluble form of TLCN can be detected in the serumand cerebrospinal fluid of patients of several neurological diseases, we are developing a sensitive ELISA
assay system to detect the soluble TLCN for monitoring the possible neuronal damage in the telencephalicregions of the patient brain.

Future Prospects
We plan to further pursue the current research
projects that are described above.
Topic (1): In addition to the current studies, wewill start a new project aiming at the elucidation ofneuronal substrate for the olfactory memory. In the
future study an emphasis will be placed on the search
for the behaviorally-relevant olfactory sensory maps
(e.g., food-odor maps) at higher olfactory centers
including the olfactory cortex and amygdala of therodent brain. An emphasis will be placed also on the
search for the neuronal mechanisms associated withodorant-induced emotional changes. These studies
will hopefully provide scientific basis for the therapeutic use of odorants and for the treatment of diseases in the olfactory nervous system.
Topic (2): We pursue to understand molecular andcellular mechanisms how the newly-generated neurons are integrated into the existing neuronal circuitand how the damaged-neurons are removed from thecircuit without damaging the function of the neuronal
circuit in the adult brain.
Topic (3): Based on our initial proteomic approaches, we now have the list of candidate molecules thatmight be involved in the olfactory axon target recognition. Systematic functional assays will be performed
to pin-down the molecular complexes that are responsible for the proper recognition of the olfactory axon
targets. The knowledge of the molecular mechanism
for target recognition in the olfactory system will beof prime importance in understanding the molecularmechanism for the formation of the neuronal circuits
in whole brain regions.
Topic (4): Cellular and molecular analysis of thecontact-mediated interaction between immune cells
and neurons will be further pursued using the co-culture assay system. In addition, we will start a systematic analysis of immune cell-mediated effects in thebrain of the TLCN-deficient mice. Collaborative
research with clinical laboratories will be pursued toestablish the methods for detecting and measuringserum TLCN for the diagnosis of possible damage oftelencephalic neurons in the brain.

Research Grants
1. Grant-in-Aid for Creative Scientific Research (JSPS)(2001-2005, ~65 millions yen / year)
2. Grant-in-Aid for Scientific Research on Priority Areas(B) (MEXT) (1999-2002, ~9.6 millions yen / year)
3. Research Grant from the Human Frontier Science
Program (1999-2001, 59000 USD / year)
4. Grant-in-Aid for Exploratory Research (MEXT)(2001-2002, 1.3 millions yen / year)
5. Research Grant from Mitsubishi Foundation (2000,
7.5 millions yen)

Select Publications (1992~2002)
1. Mori, K, Mataga, N, and Imamura, K. Differential
specificities of single mitral cells in rabbit olfactory
bulb for a homologous series of fatty acid odor molecules. J. Neurophysiol., 67, 786-789, 1992.

2. Kimura, T, Tanizawa, O, Mori, K, Brownstein, M,
and Okayama, H. Structure and expression of ahuman oxytocin receptor. Nature , 356, 526-529,
1992.
3. Tani, A, Yoshihara, Y, and Mori, K. Increase in cytoplasmic free Ca2+ elicited by noradrenalin and serotonin in cultured local interneurons of mouse olfactory bulb. Neuroscience, 49, 193-199, 1992.
4. Imamura, K, Mataga, N, and Mori, K. Coding of odor
molecules by mitral/tufted cells in rabbit olfactory
bulb: I. Aliphatic compounds. J. Neurophysiol., 68,
1986-2002, 1992.
5. Kimura, T, Azuma, C, Saji, F, Takemura, M,
Tokugawa, Y, Miki, M, Ono, M, Mori, K, andTanizawa, O. Estimation by an electrophysiologicalmethod of the expression of oxytocin receptor
mRNA in human myometrium during pregnancy. J.
Steroid Biochem. Mol. Biol., 42, 253-258, 1992.
6. Ishihara, T, Shigemoto, R, Mori, K, Takahashi, K,
and Nagata, S. Functional expression and tissue distribution of a novel receptor for vasoactive intestinal
polypeptide. Neuron, 8, 811-819, 1992.
7. Yoshihara, Y, Katoh, K, and Mori, K. Odor stimulation causes disappearance of R4B12 epitope on axonal surface molecule of olfactory sensory neurons.
Neuroscience, 53, 101-110, 1993.
8. Mochizuki-Oda, N, Negishi, M, Mori, K, and Ito, S.
Arachidonic acid activates cation channels in bovine
adrenal chromaffin cells. J. Neurochem., 61, 18821890, 1993.
9. Mori, K. Molecular and cellular properties of mammalian primary olfactory axons. Microscopy Res.
Technique, 24, 131-141, 1993.
10. Hashimoto, H, Ishihara, T, Shigemoto, R, Mori, K,
and Nagata, S. Molecular cloning and tissue distribution of a receptor for pituitary adenylate cyclase activating polypeptide. Neuron, 11, 333-342, 1993.
11. Katoh, K, Koshimoto, H, Tani, A, and Mori, K.
Coding of odor molecules by mitral/tufted cells in
rabbit olfactory bulb: II Aromatic compounds. J.
Neurophysiol., 70, 2161-2175, 1993.
12. Nemoto, Y, Ikeda, J, Katoh, K, Koshimoto, H,
Yoshihara, Y, and Mori, K. R2D5 antigen: a calciumbinding phosphoprotein predominantly expressed in
olfactory receptor neurons. J. Cell Biol., 123, 963-976,
1993.
13. Mori, K, and Shepherd, G. M. Emerging principles ofmolecular signal processing by mitral/tufted cells in
the olfactory bulb. Seminars in Cell Biol., 5, 65-74,
1994.
14. Koshimoto, H, Katoh, K, Yoshihara, Y, Nemoto, Y,
and Mori, K. Immunohistochemical demonstrationof embryonic expression of an odor receptor protein
and its distribution in the rat olfactory epithelium.
Neurosci. Lett., 169, 73-76, 1994.
15. Yoshihara, Y, Oka, S, Nagata, S, Watanabe, Y,
Kagamiyama, H, and Mori, K. ICAM-related neuronalglycoprotein, telencephalin, with brain segment-specific expression. Neuron, 12, 541-553, 1994.

37

16. Yoshihara, Y, Kawasaki, M, Tani, A, Tamada, A,
Nagata, S, Kagamiyama, H, and Mori, K. BIG-1: a
new TAG-1/F3-related member of the immunoglobulin superfamily with neurite outgrowth-promoting
activity. Neuron, 13, 415-426, 1994.
17. Mori, K, and Yoshihara, Y. Molecular recognition andolfactory processing in the mammalian olfactory system. Prog. Neurobiol., 45, 585-619, 1995.
18. Yokoi, M, Mori, K, and Nakanishi, S. Refinement ofodor molecule tuning by dendrodendritic synaptic
inhibition in the olfactory bulb. Proc. Natl. Acad. Sci.
USA, 92, 3371-3375,1995.
19. Yoshihara, Y, Kawasaki, M, Tamada, A, Nagata, S,
Kagamiyama, H, and Mori, K. Overlapping and differential expression of BIG-2, BIG-1, TAG-1, and F3:
Four members of an AxCAM subgroup of theimmunoglobulin superfamily. J. Neurobiol 28, 51-69,
1995.
20. Mori, K. Relation of chemical structure to specificityof response in olfactory glomeruli. Current Opin.
Neurobiol., 5, 467-474, 1995.
21. Hashimoto, H, Nogi, H, Mori, K, Ohishi, H,
Shigemoto, R, Yamamoto, K, Matsuda, T, Mizuno, N,
Nagata S. and Baba, A. Distribution of the mRNA fora pituitary adenylate cyclase-activating polypeptide
receptor in the rat brain: an in situ hybridization
study. J. Comp. Neurol. 371: 567-577, 1996.
22. Mizuno, T, Yoshihara, Y, Inazawza, Kagamiyama, H.
and Mori, K., cDNA cloning and chromosomal localization of the human telencephalin and its distinctive interaction with LFA-1 integrin. J. Biol. Chem.
272, 1156-1163, 1997.
23. Tian, L, Yoshihara, Y, Mizuno, T, Mori, K, andGahmberg, C G. The neuronal glycoprotein, telencephalin, is a cellular ligand for CD11a/CD18, but not
for CD11b/CD18 and CD11c/CD18 leukocyte integrins. J. Immunol. 158, 928-936, 1997.
24. Fujimori, K E, Takauji, R, Yoshihara, Y, Tamada, A,
Mori, K, and Tamamaki, N. A procedure for in situ
hybridization combined with retrograde labeling of
neurons: Application to the study of cell adhesion
molecule expression in DiI-labeled rat pyramidal
neuron. J. Histochem. and Cytochem. 45, 455-459,
1997.
25. Hino, H, Mori, K, Yoshihara, Y, Iseki, E, Akiyama, H,
Nishimura, T, Ikeda, K and Kosaka K. Reduction oftelencephalin immunoreactivity in the brain ofpatients with Alzheimer’s disease. Brain Res. 753,
353-357, 1997.
26. Yoshihara, Y, Kawasaki, M, Tamada, A, Fujita, H,
Hayashi, H, Kagamiyama, H and Mori, K. OCAM: a
new member of the neural cell adhesion moleculefamily related to zone-to-zone projection of olfactoryand vomeronasal axons. J. Neurosci. 17, 5830-5842,
1997.
27. von Campenhausen, H, Yoshihara, Y, and Mori, K.
OCAM reveals segregated mitral/tufted cell pathways
in developing accessory olfactory bulb. NeuroReport
8, 2607-2612, 1997.

38

28. Yoshihara, Y and Mori, K. Basic principles andmolecular mechanisms of olfactory axon pathfinding. Cell and Tissue Res. 290, 457-463, 1997.
29. Sugino, H, Yoshihara, Y, Copeland, N G, Gilbert, D J,
Jenkins, N A and Mori, K. Genomic organization andchromosomal localization of the mouse telencephalin
gene, a neuronal member of the ICAM family.
Genomics 43, 209-218, 1997.
30. Tamada, A, Yoshihara, Y, and Mori, K. Dendriteassociated adhesion molecule, telencephalin, promotes neurite out growth. Neuroscience Letters, 240,
163-166, 1998.
31. Benson, D, Yoshihara, Y, and Mori, K. Polarized distribution and cell -type specific localization of telencephalin, an ICAM-related neuronal cell adhesion
molecule. J. Neurosci. Res. 52, 43-53, 1998.
32. Sakurai, E, Hashikawa, T, Yoshihara, Y, Kaneko, S,
Sato, M. and Mori, K. Involvement of dendritic adhesion molecule telencephalin in hippocampal longterm potentiation. NeuroReport, 9, 881-886, 1998.
33. Mori, K, Nagao, H. and Sasaki, Y. F. Computation ofmolecular information in mammalian olfactory system. Network: Computation in Neuronal Systems, 9,79-102, 1998.
34. Yoshihara, Y, Mizuno, T, Nakahira, M, Kawasaki, M,
Watanabe, Y, Kagamiyama, H, Jishage, K, Ueda, O,
Suzuki, H, Tabuchi, K, Sawamoto, K, Okano, H,
Noda, T. and Mori, K. A genetic approach to visualization of multi-synaptic neuronal pathways using
plant lectin transgene. Neuron, 22, 33-41, 1999.
35. Arii, N, Mizuguchi, M, Mori, K. and Takashima S.
Development of telencephalin in human cerebrum.
Microscop. Res. Tech. 46, 18-23, 1999.
36. Kashiwadani, H, Sasaki, Y, Uchida, N. and Mori, K.
Synchronized oscillatory discharges of mitral/tufted
cells with different molecular receptive ranges in the
rabbit olfactory bulb. J. Neurophysiol., 82, 17861792, 1999.
37. Mori, K, Nagao. H and Yoshihara, Y The olfactory
bulb: coding and processing of odor molecule information. Science, 286, 711-715, 1999.
38. Mizuno, T, Yoshihara, Y, Kagamiyama, H, Ohsawa,
K, Imai, Y, Kohsaka, S, and Mori, K. Neuronal adhesion molecule telencephalin induces rapid cellspreading of microglia. Brain Research, 849, 58-66,
1999.
39. von Campenhausen, H. and Mori, K. Convergence ofsegregated pheromonal pathways from the accessory
olfactory bulb to the cortex. Eur. J. of Neurosci., 12,33-46, 2000.
40. Tian, L, Kilgannon, P, Yoshihara, Y, Mori, K,
Gallatin,W M, Carpen, O. and Gahmberg, C G.
Binding of T lymphocytes to hippocampal neurons
through ICAM-5 (telencephalin) and characterizationof its interaction with leukocyte integrin
CD11a/CD18. Eur. J. of Immunology, 30, 810-818,
2000.
41. Mori, K, von Campenhausen, H, and Yoshihara, Y.
Zonal organization of the mammalian main andaccessory olfactory systems. Phil. Trans. R. Soc.
Lond. B, 355, 1801-1812, 2000.
42. Yamaguchi, M, Saito, H, Suzuki, M. and Mori, K.
Visualization of neurogenesis in the central nervous
system using nestin promotor-GFP transgenic mice.
NeuroReport, 11, 1991-1996, 2000.
43. Tian, L, Nyman, H, Kilgannon, P, Yoshihara, Y, Mori,K, Anderson, L C, Kaukinen, S, Rauvala, H, Gallatin,
W M and Gahmberg, C G. Intercellular adhesion molecule-5 induces dendritic outgrowth by homophilicadhesion. J. Cell Biol., 150, 243-252, 2000.
44. Nagao, H, Yoshihara, Y, Mitsui, S, Fujisawa, H, andMori, K. Two mirror-image sensory maps withdomain organization in the mouse main olfactorybulb. NeuroReport, 11, 3023-3027, 2000.
45. Uchida, N, Takahashi, Y K, Tanifuji, M, and Mori, K.
Odor maps in the mammalian olfactory bulb: domainorganization and odorant structural features. Nature
Neurosc., 3, 1035-1043, 2000.
46. Nemoto, Y, Kearns, B G, Wenk, M R, Chen, H, Mori,K, Alb, J G Jr., De Camilli, P and Bankaitis, V A
Functional characterization of a mammalian Sac1
and mutants exhibiting substrate-specific defects in
phosphoinositide phosphatase activity. J. Biol.
Chem., 44, 34293-34305, 2000.
47. Arii N, Mizuguchi, M, Mori, K, and Takashima, S.
Ectopic expression of telencephalin in brains withholoprosencephaly. Acta Neuropathol. 100, 506-512,
2000.
48. Tatsura H, Nagao, H, Tamada, A, Sasaki, S, Kohri, K,
Mori, K. Developing germ cells in mouse testis
express pheromone receptors. FEBS letters, 488, 139144, 2001.
49. Mizuno T, Kawasaki, M, Nakahira, M, Kagamiyama,H, Kikuchi, Y, Okamoto, H, Mori, K, and Yoshihara,
Y. Molecular diversity in zebrafish NCAM family:
three members with distinct VASE usage and distinct localization. Mol. and Cell. Neurosci., 18, 119130, 2001.
50. Sawamoto K, Nakao N, Kakishita K, Ogawa Y,
Toyama Y, Yamamoto A, Yamaguchi M, Mori K,
Goldman S A, Itakura T and Okano H. Generation of
dopaminergic neurons in the adult brain from mesencephalic precursor cells labeled with a nestin-GFP
transgene. J. Neurosci., 21, 3895-3903, 2001.
51. Inaki K, Takahashi, Y K, Nagayama, S and Mori K.
Molecular-feature domains with posterrdorsalanteroventral polarity in the symmetrical sensory
maps of the mouse olfactory bulb: mapping ofodourant-induced Zif268 expression. Eur. J. of
Neurosci., 15, 1563-1574, 2002.
52. Sugitani Y, Nakai, S, Minowa, O, Nishi, M, Jishage,
K, Kawano, H, Mori, K, Ogawa, M, and Noda, T. Brn1 and Brn-2 share crucial roles in the production andpositioning of mouse neocortical neurons. Genes andDevelopment, 16, 1760-1765, 2002.
53. Nagao H, Yamaguchi, M, Takahashi, Y, and Mori, K.
Grouping and representation of odorant receptors indomains of the olfactory bulb sensory map.
Microscopy Res. & Technique 58, 168-175, 2002.

39

Department of
Neurophysiology
Outline and Research Objectives
Our laboratory was founded in 1953 as a Department of Brain Physiology in Institute forBrain Research, University of Tokyo Faculty of Medicine, and in 1996 integrated into
University of Tokyo Graduate School of Medicine. We have been teaching Neurophysiology
to undergraduate students in Medical School, and Master and PhD course students. As forresearch, using patch clamp techniques in combination with molecular techniques, we aim
at elucidating cellular and molecular mechanisms underlying synaptic transmission andmodulation. We are particularly interested in dynamic changes at CNS synapses during
postnatal development.
Synapses in the CNS play pivotal roles in neuronal integration and plasticity. During
ontogeny CNS synapses undergo protein reformations, thereby establishing mature and differentiated synaptic functions. Synapses also undergo changes in response to electrical andchemical stimuli. Various proteins in the presynaptic and postsynaptic cells are involved insuch changes, and it is important to determine the role of these proteins in synaptic functions and modulations. To this end, we combine electrophysiological, molecular and morphological techniques to synapses visually identified in the CNS slices. In the rodent auditory brainstem there is a giant excitatory synapse called the calyx of Held. At this synapse it
is possible to make simultaneous whole-cell recordings from a presynaptic terminal and apostsynaptic target cell (Fig. 1). Furthermore, various molecules can be loaded into a nerve
terminal via whole-cell pipette perfusion during the recording (Fig.2). The large structure ofthis synapse also allows us to identify presynaptic proteins immunocytochemically, andeven to follow translocation of presynaptic molecules upon stimulation. With theseapproaches in combination with genetic manipulations and protein overexpressions, we
aim at addressing the molecular basis underlying synaptic functions.

Fig. 1 Simultaneous pre- and postsynaptic wholecell recordings at the calyx of Held
synapse. Inset records on the left show a presynaptic action potential and excitatory postsynaptic
currents (EPSC). Righthand inset illustrates theprocedure of patch-clamp whole-cell recording
method.

Fig. 2 Illustration of pipette perfusion system. A
tube pulled from Eppendorf yellow tip is inserted
into the patch pipette with its tip about 600 mm
from the tip of the patch pipette.

40Faculties and Students
Professor and Chair

Tomoyuki Takahashi MD, PhD(appointed since 1993)
Lecturer
Tetsuhiro Tsujimoto MD, PhD
Associate .................................1
Postdoctral Fellow ..................2
Graduate Students ..................9Technical Assistant ................1
Secretary .................................2

Past Research and Major Accomplishments
1. In simultaneous pre- and postsynaptic recordings
at the calyx of Held synapse, we identified the target of presynaptic inhibition via metabotropic glutamate receptors and GABAB receptors as being
the voltage-dependent Ca2+ channel (Takahashi etal., Science 1996, J Neurosci,1998), and that βγ
subunits of the trimeric G protein Go mediate thepresynaptic inhibition (Kajikawa et al, 2001
PNAS).
2. We identified that phorbol ester stimulates exocytotic machinery downstream of Ca2+ influx thereby
causing synaptic facilitation. By loading inhibitory
peptides into the nerve terminal, we also clarified
that both εPKC (Fig. 4a) and the Munc13-Doc2αinteraction (Fig. 4b) underlie the phorbol esterinduced facilitatory mechanism (Hori et al., J
Neurosci 1998; Saitoh et al, 2002).
3. We found that Ca2+ channel types mediating CNS
synaptic transmission switch during development(Iwasaki et al, J Neurosci 2000).

Fig. 3 Presynaptic inhibitory effect of baclofen occludes that of
L-AP4, suggesting that GABAB and metabotropic glutamate
receptors share a common intracellular mechanism for presynaptic inhibition. Righthand illustration summarizes our current
conclusion. Adenosine A1 receptors also share the same mechanism (Kimura and Takahashi, unpublished observeation).

Fig. 4 Left column, Phorbol etser-induced EPSC potentiation isattenuated by the PKC inhibitor BIS (A) or PKC inhibtitory peptide directly loaded into the calyceal terminal (B). Right column,
The Mid peptide, which interferes with the Doc2α-Munc13-1
interaction attenuated the phorbol etser-induced EPSC potentiation.

4. By loading G protein-related compounds into thecalyx of Held nerve terminal, we clarified that themain role of presynaptic G proteins is to accelerate
the replenishment of synaptic vesicles depleted
after massive release (Takahashi et al. Science2000).
5. We have demonstrated that the quantal analysis
established at the neuromuscular junction is applicable to the CNS synapse (Sahara & Takahashi, J
Physiol 2001).
6. We found that NMDA receptors are unfavorable
for the reliability of fast synaptic transmission,and that their developmental decline regulated byauditory activity at the calyx of Held synapse differentiates the synapse into the high-fidelity one(Futai et al., J Neurosci 2001)

Fig. 5 Synaptic currents mediated by presynaptic N-type Ca2+
channels at 5 different central synapses. At 3 synapses, contribution of N-type Ca2+ channels to synaptic transmission decline
and disappear with postnatal development, whereas at other 2
synapses it remained unchanged throughout development.

7. We found that presynaptic Ca2+ currents undergo
facilitation by repetitive stimulation (Forsythe etal, 1998 Neuron) via acceleration of their gating
kinetics (Cuttle et al., 1998 J Physiol) and that this41

facilitation is mediated by the Ca2+ binding protein
NCS-1 (Tsujimoto et al, 2002 Science).
8. We found that cytoplasmic glutamate concentration in the nerve terminal directly affects thevesicular content of glutamate and that postsynaptic AMPA and NMDA receptors are not saturated
by a single packet of vesicular transmitter(Ishikawa et al, 2002 Neuron).
Fig. 8 Switching the L-glutamate concentration in the presynaptic whole-cell pipette from 1 mM to 100 mM markedly potentiated both quantal (blue) and evoked (red) EPSCs at the calyx ofHeld synapse at physiological temperature.

Current Research

Fig. 6 Postsynaptic action potentials in response to presynaptic
stimulation at 100 Hz at the developing calyx of Held synapse inmice. As mice mature fidelity of transmission increases, whereas blocking NMDA receptors using APV increases the fidelity at
all ages.

1. Identification of presynaptic potassium channels
involved in regulation of transmitter release.
2. Clarification on the developmental changes in thequantal synaptic responses.
3. Mechanism and roles of presynaptic adenosine
receptors.
4. Developmental changes in the size of presynaptic
Ca2+ domain.
5. Identification in the target protein downstream of
εPKC.
6. Clarification in the different properties between
presynaptic N-type and P/Q-type Ca2+ channels.

Future Prospects
1. To elucidate the minimal essential molecules
required for short-term and long-term synaptic
plasticity.
2. To clarify the molecular basis underlying thedevelopmental speeding in the excitatory synaptic
currents.
3. To clarify the presynaptic factor(s) determining
the release probability.

Research Grants (in the past 5 years)
1996-2000
Research for the Future Program by the Japan Societyof Promotion of Sciences “Molecular mechanisms ofcentral synaptic modulation underlying the memory
formation” ¥ 317,092,000
2001-2002
Grant-in-Aid for Specially promoted Research from
the Ministry of Education, Culture, Sports, Scienceand Technology “Cellular and molecular mechanisms
underlying postnatal development and differentiation of central synapses” ¥216,000,000
Fig. 7 The NCS-1 C-terminal peptide, when loaded into thecalyx of Held presynaptic terminal, blocked the activity-dependent facilitations of presynaptic P/Q-type Ca2+ currents in a paired
pulse protocol (A) and in a tetanic stimulation (100 Hz) protocol(C). The peptide had no effect on the current voltage relationship of presynaptic Ca2+ currents.

42

Select Publications
1. Ishikawa T, Sahara, Y, Takahashi T. A single packetof transmitter does not saturate postsynaptic glutamate receptors. Neuron 34, 613-621, 2002.
2. Tsujimoto T, Jeromin A, Saitoh N, Roder JC and
Takahashi T Neuronal calcium sensor 1 and activitydependent facilitation of P/Q-type calcium currents
at presynaptic nerve terminals. Science 275,22762279, 2002.
3. Saitoh N., Hori T. & Takahashi T. Activation of theepsilon isoform of protein kinase C in the mammalian nerve terminal. Proc. Natl. Acad. Sci. USA.
98,14017-14021, 2001.
4. Sahara Y. & Takahashi T. Quantal components of theexcitatory postsynaptic currents at a rat central auditory synapse. J. Physiol. (London) 536,189-197, 2001.
5. Iwasaki S. & Takahashi T. Developmental regulationof transmitter release at the calyx of Held in rat auditory brainstem. J.Physiol. (London) 534,861-871,
2001.
6. Kajikawa Y., Saitoh N.& Takahashi T. GTP-binding
protein βγ subunits mediate presynaptic calcium current inhibition by GABAB receptor. Proc. Natl. Acad.
Sci.U.S.A 98,8054-8058, 2001.
7. Ishikawa T. & Takahashi T. Mechanisms underlying
presynaptic facilitatory effect of cyclothiazide at thecalyx of Held of juvenile rats. J. Physiol. (London)
533, 423-431, 2001.
8. Futai K., Okada M., Matsuyama K. & Takahashi T.
High-fidelity transmission acquired via a developmental decrease in NMDA receptor expression at anauditory synapse. J. Neurosci. 21,3342-3349, 2001.
9. Yamauchi, T., Hori T. & Takahashi T. Presynaptic
inhibition by muscimol through GABAB receptors.
Eur. J. Neurosci. 12, 3433-3436, 2000.
10. Takahashi T., Hori T., Kajikawa Y.& Tsujimoto T.
The role of GTP-binding protein activity in fast central synaptic transmission. Science 289, 460-463,
2000.
11. Okada M., Onodera K., Van Renterghem C., Sieghart
W.& Takahashi T. Functional correlation of GABAA
receptor α subunits expression with the properties
of IPSCs in the developing thalamus. J. Neurosci. 20,
2202-2208, 2000.
12. Iwasaki S., Momiyama A., Uchitel O.D.& Takahashi
T. Developmental changes in calcium channel types
mediating central synaptic transmission. J. Neurosci.
20, 59-65, 2000.
13. Hori T., Takai Y. & Takahashi T. Presynaptic mechanism for phorbol ester- induced synaptic potentiation. J. Neurosci. 19, 7262-7267, 1999.
14. Terada S., Tsujimoto T., Takei Y., Takahashi T. &Hirokawa N. Impairement of inhibitory synaptictransmission in mice lacking synapsin 1. J. Cell Biol.
145, 1039-1048, 1999.
15. Kobayashi K., Manabe T. & Takahashi T. Calciumdependent mechanisms involved in presynaptic
long-term depression at the hippocampal mossy

fibre-CA3 synapse. Eur. J. Neurosci. 11, 1633-1638,
1999.
16. Mori H., Manabe T., Watanabe M., Satoh Y., SuzukiN., Toki S., Nakamura K., Yagi T., Kushiya E.,
Takahashi T., Inoue Y., Sakimura K. & Mishina M.
Role of the carboxyl-terminal region of the GluR ε2subunit in synaptic localization of the NMDA receptor channel. Neuron 21, 571-580, 1998.
17. Manabe T., Noda Y., Mamiya T., Katagiri H., HoutaniT., Nishi M., Noda T., Takahashi T., Sugimoto T.,
Nabeshima T. & Takeshima H. Facilitation of longterm potentiation and memory in mice lacking nociceptin receptors. Nature 394, 577-581, 1998.
18. Cuttle M. F, Tsujimoto T., Forsythe I.D. & Takahashi
T. Facilitation of the presynaptic calcium current at
an auditory synapse in rat brainstem. J.
Physiol.(London) 512, 723-729, 1998.
19. Iwasaki S. & Takahashi T. Developmental changes incalcium channel types mediating synaptic transmission in rat auditory brainstem. J. Physiol.(London)
509, 419-423, 1998.
20. Takahashi T., Kajikawa Y. & Tsujimoto T. G-proteincoupled modulation of presynaptic calcium currents
and transmitter release by a GABA B receptor. J.
Neurosci. 18, 3138-3146, 1998.
21. Forsythe I.D., Tsujimoto T., Barnes-Davies M., Cuttle
M.F., & Takahashi T. Inactivation of presynaptic calcium current contribute to synaptic depression at afast central synapse. Neuron 20, 797-807, 1998.
22. Takahashi T., Forsythe I., Tsujimoto T., BarnesDavies M. & Onodera K. Presynaptic calcium current
modulation by a metabotropic glutamate receptor.
Science 274, 594- 597, 1996.
23. Kobayashi K., Manabe T. & Takahashi T. Presynaptic
long-term depression at the hippocampal mossy
fiber-CA3 synapse. Science 273, 648-650, 1996.
24. Yokoi M., Kobayashi K., Manabe T., Takahashi T.,
Sakaguchi I., Katsuura., Shigemoto R., Ohishi H.,
Nomura S., Nakamura K., Nakao K., Katsuki M., &Nakanishi S. Impairment of hippocampal mossy
fiber LTD in mice lacking metabotropic glutamate
receptor subtype 2. Science 273, 645-647, 1996.
25. Takahashi T., Feldmeyer D., Suzuki N., Onodera K.,
Cull-Candy S.G., Sakimura K., & Mishina M.
Functional Correlation of NMDA receptor ε subunits
expression with the properties of single-channel andsynaptic current in the developing cerebellum. J.
Neurosci. 16, 4376-4382, 1996.
26. Silver R.A., Cull-Candy S.G. & Takahashi T. NonNMDA glutamate receptor occupancy and open
probability at a rat cerebellar synapse with single
and multiple release sites. J. Physiol. (London) 494,
231-250, 1996.
27. Kutsuwada T., Sakimura K., Manabe T., Takayama
C., Katakura N., Kushiya E., Natsume R., Watanabe
M., Inoue Y., Yagi T., Aizawa S., Arakawa M.,
Takahashi T., Nakamura Y., Mori H. & Mishina M.
Impairment of suckling response, trigeminal neuronal pattern formation and hippocampal LTD in

43

NMDA receptor ε2 subunit mutant mice. Neuron 16,
333-344, 1996.
28. Farrant M., Feldmeyer D., Takahashi T. & CullCandy S.G. NMDA-receptor channel diversity in thedeveloping cerebellum. Nature 368, 335-339, 1994.
29. Momiyama A. & Takahashi T. Calcium channels
responsible for potassium-induced transmitter
release at rat cerebellar synapses. J Physiol. (London)
476, 197-202, 1994.
30. Takahashi T. & Momiyama A. Different types of calcium channels mediate central synaptic transmission. Nature 366,156-158, 1993.

44

Department of Cellular andMolecular Pharmacology
Outline and Research Objectives
Our department was founded in 1885 as the first pharmacology department in Japan. It
has had a strong research background in the field of calcium (Ca2+) signalling since Professor
Emeritus Setsuro Eabashi discovered in 1950’s the regulatory role of intracellular Ca2+ concentration in muscle contraction. Since then the field of Ca2+ signalling research has expanded extensively: the Ca2+ signal is now known as a molecular switch in a vast array of important cell functions including muscle contraction, exocytosis, cell proliferation, immune
responses and regulation of synaptic functions. The present research group led by Professor
Iino is interested in the general principle of the Ca2+ signalling mechanism and is particularlyinterested in Ca2+ signalling in neurons and muscle cells at present. We have recently
expanded the scope of our research to various signaling molecules upstream and downstream of Ca2+ signals. A notable feature of our department is that we have had an assemblyof staff members with diverse backgrounds, e.g., cell physiology, molecular biology and neurobiology. We believe that the collaboration of these people will facilitate the elucidation of
the medical and biological significance of Ca2+ signalling and related subjects.

Faculties and Students
Professor

Masamitsu Iino, M.D., Ph.D.
(From April 1995)
Associate Professor Kenzo Hirose, M.D., Ph.D.
Associate .................................2
Postdoctoral Fellows...............3
Graduate Students ................11
Research Students...................3
Laboratory Staff ......................3

Past Research and Major Accomplishments
Our research focuses on the regulation of Ca2+ signals. In particular, we have made important contributions to clarifying intracellular Ca2+ release mechanisms. Our research also includes signaling molecules
upstream and downstream of Ca2+ signals. Throughthese works, we have clarified feedback mechanismsthat are essential to the generation of spatiotemporal
patterns of Ca2+ signals.

1) Functional properties of IP3R.
The receptor-mediated activation of phospholipase
C results in the production of inositol 1,4,5-trisphosphate (IP3), which then releases Ca2+ from the intracellular Ca2+ store. This Ca2+ mobilization is a central
mechanism in the generation of Ca2+ signals for the
regulation of various cell functions. We showed forthe first time that the Ca2+ release via IP3R is activatedby submicromolar concentrations of Ca2+ (Fig. 1). We
also discovered an inhibitory role of Ca2+ at higher
concentrations. Thus, IP 3R activity is biphasicallydependent on cytosolic Ca2+ concentration with the
maximum level of activation obtained at 0.3 µM (J.
Gen. Physiol., 1990). We also showed that the effectsof Ca2+ on IP3R activity have no notable delay in experiments using both caged IP3 and caged Ca2+ (Nature,1992). These results suggest the presence of a positive
feedback loop in IP3R-mediated Ca2+ release. Such a
positive feedback loop is expected to have a major
effect on Ca2+ release kinetics via IP3R (Nature, 1994b).
This notion, which is very popular in the field of Ca2+
signalling, now has a strong molecular basis as shown
below.
We then studied the structure-function relationship of IP3R. IP3R consists of three subtypes (IP3R1,
IP3R2 and IP3R3), that are encoded by different genes
and are expressed in a tissue- and development-specific manner. In collaboration with Prof. Kurosaki of
Kansai Medical University, we genetically engineered
DT40 B cells to express only one IP3R subtype andFigure 1. IP3 and Ca2+ dependence of IP3R activity. Both IP3 andCa2+ are required in the activation of IP3R.

45

showed that each IP3R subtype has distinct properties(EMBO J., 1999). For example, the order of IP3 sensitivity is IP3R2 > IP3R1 >I P3R3. Furthermore, we succeeded in mapping the Ca2+ sensor region of IP3R1 at
glutamate 2100 (Fig. 2) (EMBO J., 2001). Substitutionof this amino acid by aspartate (E2100D) resulted in a
10-fold decrease in the Ca2+ sensitivity of IP3R1. When
we expressed the E2100D mutant IP3R in DT40 cells,
the rates of increase of Ca2+ concentration and Ca2+
oscillations during an agonist-induced response weresignificantly reduced compared with cells expressing
wild-type IP3R1 (Fig. 3). These results demonstratethat the Ca2+-mediated feedback regulation of IP3R
activity is extremely important for the generation of

Figure 2. Ca2+ sensor of IP3R. Glutamate (E) at position 2100
functions as the Ca2+ sensor of IP3R.

Figure 3. Ca2+ signalling in cells expressing mutant (E2100D)
IP3R with reduced Ca2+ sensitivity. BCR-mediated Ca2+ oscillation was abolished in cells expressing the mutant IP3R.

Figure 4. Positive feedback regulation of IP3R via the Ca2+ sensor. Ca2+-sensor-mediated positive feedback regulation of Ca2+
release via IP3R is essential for the spatio-temporal pattern generation of agonist-stimulated Ca2+ release.

46

spatiotemporal patterns of Ca2+ signals (Fig. 4).

2) Ca2+ imaging using intact tissues.
In order to fully understand the physiological functions of Ca2+-mediated cell signalling mechanisms, it isimportant to study Ca2+ signalling in cells that communicate with each other within intact tissues. With
this notion in mind, we performed Ca 2+ imaging
experiments using intact vascular tissues. Thismethod successfully revealed that vascular smoothmuscle cells respond to sympathetic nerve stimuli
with Ca2+ waves and oscillations (EMBO J., 1994). To
our surprise, resting arterial smooth muscle cells were
not quiescent in terms of Ca2+ signals. We discovered
spontaneous Ca2+ transients in unstimulated vascularsmooth muscle cells (J. Physiol., 1999). These Ca2+transients are called “Ca2+ ripples” because their peak
amplitudes were much smaller than those of sympathetic nerve-stimulated Ca2+ responses. Ca2+ ripples
contribute to some extent to the resting tonus of thevascular wall. Other physiological functions await further clarification.
We have now extended the application of the tissue imaging method to intestinal tissues, and have forthe first time succeeded in simultaneous imaging of
Ca2+ signals in smooth muscle cells and interstitialcells of Cajal, which are the putative intestinal pacemaker cells (J. Physiol., 2002). This method may help
us elucidate the rhythm-making mechanism in intestinal tissues.
3) Excitation-contraction coupling and malignant
hyperthermia.
In skeletal muscle cells depolarization of the plasma membrane induces Ca2+ release via the Ca2+ releasechannel (ryanodine receptor, RyR) on the adjacent sarcoplasmic reticulum. This process is called excitationcontraction (EC) coupling and has been studied using
various methods. We have made several important
contributions to clarifying the molecular basis of E-C
coupling. There are three subtypes of RyR (RyR1,
RyR2 and RyR3) encoded by different genes. First, we
showed that RyR1 is critically important for EC coupling (Nature, 1994a). Second, we showed that RyR1
but not RyR2 or RyR3 supports the skeletal muscle EC
coupling (EMBO J., 1996). Third, a stretch of about
100 amino acids (D2 region), which is ~1,300 aminoacids removed from the N terminus of RyR1, is important for EC coupling ( J. Biol. Chem. , 1997). These
results provided a framework for the molecular elucidation of EC coupling.
In relation to this subject, we have identified oneof the genetic causes of malignant hyperthermia, acomplication of general anesthesia with a life-threatening fever, resulting from an abnormality in theskeletal muscle calcium release channel, RyR1. We

identified a point mutation (L4838V) that is responsible for the gain-of-function mutation of RyR1 in
malignant hyperthermia patients with marked clinicalsymptoms (Jpn. J. Pharmacol., 2002).
We also discovered junctophilin, a protein that is
responsible for the junction formation between theplasma membrane and the sarcoplasmic reticulum
membrane (Mol. Cell, 2000).

4) IP3 imaging and discovery of a new IP3 signallingmechanism.
We have succeeded in visualizing the changes inintracellular IP3 concentration using the translocationof the GFP-tagged PH domain of PLC-δ1 (GFP-PHD)(Science, 1999). Using this method, we showed for the
first time the dynamic changes in the intracellular IP3
concentration (IP3 oscillations and waves) (Fig. 5).
These results show that the IP3 dynamics is an important factor for the generation of dynamic spatiotemporal patterns of Ca2+ signals. Furthermore, we applied
this method to measuring the changes in IP3 concentration in cerebellar Purkinje cells and found a novel
Ca2+-mediated IP3 signalling pathway that leads to IP3
production following climbing fiber inputs into
Purkinje cells (Neuron, 2001) (Fig. 6).

Current Research
Based on the results of our studies of Ca 2+ signalling, we recognize the importance of the spatiotemporal distribution of signalling molecules in defining
cell signals. Thus, global Ca2+ waves and oscillations insmooth muscle cells result in contraction of the cells,
while localized, transient rises at subplasmalemmal
regions (Ca2+ sparks) result in relaxation of the cells.
We, therefore, believe that it is extremely importantto visualize the spatiotemporal distribution of signalling molecules within intact cells. To that end, weare currently engaged in the development of indicators of important signalling molecules (see below).
Two types of excitable cells, neurons of the central
nervous system and smooth muscle cells, are our current target cells. In particular, neurons have distinct
cell polarity, therefore the spatiotemporal distributionof signalling molecules should be of great importancefor the functions of neurons.

1) Development of novel signal indicators.
We are currently involved in the development of
new genetically coded indicators of various importantcell signals including nitric oxide (NO), protein phosphorylation and a Ca2+-dependent transcription factor.
Some of the indicators are now being expressed incells for analysis, and others are now close to application to cells.

2) Ca2+ signalling in Purkinje cells and synaptic
plasticity.
Ca2+ and IP3 dynamics in Purkinje cells are now
studied in Purkinje cells in conjunction with themolecular basis of synaptic plasticity. We use bothconfocal and two-photon excitation microscopies toobserve signals in individual dendrites.
3) Molecular approaches to the study of IP3Rmediated Ca2+ signalling.
We have identified the Ca2+ sensor region of IP3R
and showed that a mutation at this site resulted in
the inhibition of Ca2+ signals. We are introducing thismutation to various cells to inhibit Ca 2+ signals.
Through this approach it will be possible to clarify theroles of Ca2+ signals in various cells.

Future Prospects
1) Elucidation of the relationship between Ca2+
signalling and cell functions.
Intracellular Ca2+ signals exhibit extremely dynamic changes both temporally and spatially. Such property allows the Ca2+ signal to be an extremely versatileswitch regulating diverse cell functions; from transmitter release in neurons to cell proliferation, andfrom muscle contraction to apoptosis. The role of Ca2+
signals in skeletal muscle contraction has been thoroughly clarified. However, there are still many cellfunctions in which Ca2+ signals are thought to play
important regulatory roles, but their mechanisms
remain elusive. We would like to elucidate the relationship between the Ca2+ signalling mechanism andthe regulation of cell functions in all such frontiers.
2) Visualization and analysis of molecular events
at synapses during synaptic plasticity.
Among many cell functions, we are particularlyinterested in those of neurons, which have such aunique cell polarity and provide an excellent platform
for spatiotemporal cell signalling. We are interested incell signals upstream and downstream of the Ca2+ signals. Combining new signal indicators and imaging
methods, we wish to visualize the spatiotemporal distribution of cell signals in order to elucidate molecular
events at the synapses during synaptic plasticity that
underlies learning and memory.

Research Grants
1. Core Research for Evolutional Science andTechnology (CREST) from Japan Science andTechnology Cooperation “Calcium SignallingResearch with Advancement of Imaging and47

Molecular Genetic Methods“ (Years 1996-2000)¥676,312,000.
2. Grant-in-Aid for Scientific Research (Specially
Promoted Research) from the Minisitry of Education,
Science, Sports and Culture. “Visualization of
Intracellular Signal Flow” (Years 2000-2004)¥430,000,000.

Select Publications (Reprints of the ten references with asterisks are attached.)
1. Iino, M. Tension responses of chemically skinned
fibre bundles of the guinea-pig taenia caeci undervaried ionic environments. J. Physiol. 320, 449-467,
1981.
2. Iino, M. Calcium dependent inositol trisphosphateinduced calcium release in the guinea-pig taenia
caeci. Biochem. Biophys. Res. Commun. 142, 47-52,
1987.
3. Iino, M. Calcium induced calcium release mechanism
in guinea-pig taenia caeci. J. Gen. Physiol. 94, 363383, 1989.
4.* Iino, M. Biphasic Ca2+ dependence of inositol 1,4,5trisphosphate-induced Ca release in smooth muscle
cells of the guinea pig taenia caeci. J. Gen. Physiol.
95, 1103-1122, 1990.
5. Iino, M. Effects of adenine nucleotides on inositol
1,4,5-trisphosphate-induced calcium release in vascular smooth muscle cells. J. Gen. Physiol. 98, 681-698,
1991.
6.* Iino, M. and Endo, M. Calcium-dependent immediate
feedback control of inositol 1,4,5-trisphosphateinduced Ca2+ release. Nature 360, 76-78, 1992.
7. Oyamada, H., Iino, M. and Endo, M. Effects of ryanodine on the properties of Ca2+ release from the sarcoplasmic reticulum in skinned skeletal muscle fibresof the frog. J. Physiol. 470, 335-3348, 1993.
8. Iino, M., Yamazawa, T., Miyashita, Y., Endo, M. andKasai, H. Critical intracellular Ca2+ concentration for
all-or-none Ca 2+ spiking in single smooth muscle
cells. EMBO J. 12, 5287-5291., 1993
9. Tsukioka, M., Iino, M. and Endo, M. pH dependence
of inositol 1,4,5-trisphosphate-induced Ca2+ release inpermeabilized smooth muscle cells of the guinea-pig.
J. Physiol. 475, 369-375, 1994.
10. Oyamada, H., Murayama, T., Takagi, T., Iino, M.,
Iwabe, N., Miyata, T. Ogawa, Y. and Endo, M.
Primary structure and distribution of ryanodinebinding protein isoforms of the bullfrog skeletal
muscle. J. Biol. Chem. 269, 17206-17214, 1994.
11. Kasai, Y., Iino, M., Tsutsumi, O., Taketani, Y. andEndo, M. Effects of cyclopiazonic acid on rhythmic
contractions in uterine smooth muscle bundles of
the rat. Br. J. Pharmacol. 112, 1132-11136, 1994.
12.* Takeshima, H., Iino, M., Takekura, H., Nishi, M.,
Kuno, J., Minowa, O., Takano, H. and Noda, T.
Excitation-contraction uncoupling and musculardegeneration in mice lacking functional skeletal muscle ryanodine-receptor gene. Nature 369, 556-559,
1994.

48

13.* Iino, M., Kasai, H. and Yamazawa, T. Visualizationof neural control of intracellular Ca2+ concentration insingle vascular smooth muscle cells in situ. EMBO J.
13, 5026-5031, 1994.
14.* Hirose, K. and Iino, M. Heterogeneity of channel
density in inositol-1,4,5-trisphosphate- sensitive Ca2+
stores. Nature 372, 791–794, 1994.
15. Kasai, Y., Tsutsumi, O., Taketani, Y., Endo, M. andIino, M. Stretch-induced enhancement of contractions in uterine smooth muscle of rats. J. Physiol.
486, 373-384, 1995
16. Takeshima, H., Yamazawa, T., Ikemoto, T.,
Takekura, H., Nishi, M., Noda, T. and Iino, M. Ca2+induced Ca2+ release in myocytes from dyspedic micelacking type-1 ryanodine receptor. EMBO J. 14,
2999–3006, 1995.
17. Ikemoto, T., Iino, M. and Endo, M. Enhancing effectof calmodulin on Ca2+-induced Ca2+ release in the sarcoplasmic reticulum of rabbit skeletal muscle fibres.
J. Physiol. 487, 573-582, 1995.
18. Ikemoto, T., Iino, M. and Endo, M. Effect of calmodulin antagonists on calmodulin-induced biphasic
modulation of Ca 2+ -induced Ca 2+ release. Br. J.
Pharmacol. 118, 690-694, 1996.
19. Takeshima, H., Ikemoto, T., Nishi, M., Nishiyama,N., Shimuta, M., Sugitani, Y., Kuno, J., Saito, I., Saito,
H., Endo, M., Iino, M. and Noda, M. Generation andcharacterization of mutant mice lacking ryanodine
receptor type 3. J. Biol. Chem. 271, 19649-19652,
1996.
20.* Yamazawa, T., Takeshima, H., Sakurai, T., Endo, M.
and Iino, M. Subtype specificity of ryanodine receptor for Ca2+ signal amplification in excitation-contraction coupling. EMBO J. 15, 6172-6177, 1996.
21. Yamazawa,T., Takeshima, H., Shimuta, M. and Iino,
M. A region of the ryanodine receptor critical for
excitation-contraction coupling in skeletal muscle. J.
Biol. Chem. 272, 8161-8164, 1997.
22. Ikemoto, T., Komazaki, S., Takeshima, H., Nishi, M.,
Noda, T., Iino, M. and Endo, M. Functional and morphological features of skeletal muscle from mutant
mice lacking both type 1 and type 3 ryanodine receptors. J. Physiol. 501, 305-312, 1997.
23. Kasai, Y., Yamazawa, T., Sakurai, T., Taketani, Y.
and Iino, M. Endothelium-dependent frequency
modulation of Ca2+ signaling in individual vascularsmooth muscle cells of the rat. J. Physiol. 504, 349357, 1997.
24. Takesako, K., Sasamoto, K., Ohkura, Y., Hirose, K.
and Iino, M. Low-affinity fluorescent indicator for
intracellular calcium ions. Anal. Commun. 34, 391392, 1997.
25. Hirose, K., Kadowaki, S. and Iino, M. Allosteric regulation by cytoplasmic Ca2+ and IP3 of the gating of IP3
receptors in guinea pig vascular smooth muscle cells.
J. Physiol. 506, 407-414, 1998.
26. Takeshima, H., Komazaki, S., Hirose, K., Nishi, M.,
Noda, T. and Iino, M. Embryonic lethality andabnormal cardiac myocytes in mice lacking ryanodine receptor type 2. EMBO J. 17, 3309-3316, 1998.

27. Takeshima, H., Shimuta, M., Komazaki, S., Ohmi, K.,
Nishi, M., Iino, M., Miyata, A. and Kangawa, K.
Mitsugumin29, a novel synaptophysin family member from the triad junction in skeletal muscle.
Biochem. J. 331, 317-322, 1998.
28. Okada, H., Bolland, S., Hashimoto, A., Kurosaki, M.,
Kabuyama, Y., Iino, M., Ravech, J.V., Kurosaki, T.
Role of the inositol phosphatase SHIP in B cell receptor-induced Ca2+ oscillatory response. J. Immunol.
161, 5129-5132, 1998.
29. Ikemoto, T., Takeshima, H. Iino, M. and Endo, M.
Effect of calmodulin on Ca2+-induced Ca2+ release ofskeletal muscle from mutant mice expressing either
ryanodine receptor type 1 or type 3. Pflügers Arch.
437, 43-48, 1998.
30. Manabe, T., Noda, Y., Mamiya, T., Katagiri, H.,
Houtani, T., Nishi, M., Noda, T., Takahashi, T.,
Sugimoto, T., Nabeshima, T. and Takeshima, H.
Facilitation of long-term potentiation and memory inmice lacking nociceptin receptor. Nature 394, 577581, 1998.
31. Hirabayashi, T., Kume, K., Hirose, K., Yokomizo, T.,
Iino, M., Itoh, H. and Shimizu, T. Critical duration of
intracellular Ca2+ response required for continuoustranslocation and activation of cytosolic phospholipase A2. J. Biol. Chem. 274, 5163-5169, 1999.
32.* Miyakawa, T., Maeda, A., Yamazawa, T., Hirose, K.,
Kurosaki, T. and Iino, M. Encoding of Ca2+ signals bydifferential expression of IP 3 receptor subtypes.
EMBO J. 18, 1303-1308, 1999.
33. Hashimoto, A., Hirose, K., Okada, H., Kurosaki, T.
and Iino, M. Inhibitory modulation of B cell receptormediated Ca2+ mobilization by SH2 domain containing inositol 5’-phosphatase (SHIP). J. Biol. Chem. 274,
11203-11208, 1999.
34. Hirose, K., Takeshima, H. and Iino, M. Fluorescentindicators for inositol 1,4,5-triphosphate based on
bioconjugates of pleckstrin homology domain andfluorescent dyes. Anal. Commun. 36, 175-177, 1999.
35.* Hirose, K., Kadowaki, S., Tanabe, M., Takeshima,
H. and Iino, M. Spatio-temporal dynamics of Inositol
1,4,5-trisphosphate that underlies complex Ca 2+
mobilization patterns. Science 284, 1527-1530, 1999
36. Asada, Y, Yamazawa, T., Hirose, K., Takasaka, T.
and Iino, M. Dynamic signaling in rat arterial smoothmuscle cells under the control of local reninangiotensin system. J. Physiol. 521, 497-505, 1999.
37. Nishi, M. Komazaki, S., X Kurebayashi, N., Ogawa,Y., Noda, T., Iino, M. and Takeshima, H. Abnormalfeatures in skeletal muscle from mice lacking
Mitsugumin29. J. Cell Biol. 147, 1473-1480, 1999
38. Takeshima, H., Komazaki, S., Nishi, M., Iino, M. andKangawa, K. Junctophilins: a novel family of junctional membrane complex proteins. Molecular Cell,6, 11-22, 2000.
39. Hashimoto, A., Hirose, K., Kurosaki, T. and Iino, M.
Negative control of store-operated Ca2+ influx by B
cell receptor crosslinking. J. Immunol. 166, 10031008, 2001.

40. Nemoto, T., Kimura, R., Ito, K., Tachikawa, A.,
Miyashita, Y., Iino, M. and Kasai, H. Sequentialreplenishment mechanism of exocytosis in pancreatic acini. Nature Cell Biol. 3, 253-258, 2001.
41. Inoue, T., Kikuchi, K., Hirose, K., Iino, M. andNagano, T. Small molecule-based laser inactivation of
inositol 1,4,5-trisphosphate receptor. Chem. Biol. 8,9-15, 2001.
42.* Miyakawa, T., Mizushima, A., Hirose, K.,
Yamazawa, T., Bezprozvanny, I., Kurosaki, T. andIino, M. Ca2+-sensor region of IP3 receptor controls
intracellular Ca2+ signalling. EMBO J. 20, 1674-1680,
2001.
43. Fujiwara, A., Hirose, K., Yamazawa, T. and Iino, M.
Reduced IP3 sensitivity of IP3 receptor in Purkinje
neurons. NeuroReport 12, 2647-2651 2001.
44.* Okubo, Y., Kakizawa, S., Hirose, K. and Iino, M.
Visualization of IP3 dynamics reveals a novel AMPA
receptor-triggered IP3 production pathway mediatedby voltage-dependent Ca2+ influx in Purkinje cells.
Neuron 32, 113-122, 2001.
45. Namikis, S., Hirose, K. and Iino, M. Mapping ofheme-binding domains in soluble guanylyl cyclase
β1 subunit. Biochem. Biophys. Res. Commun. 288,
798-804, 2001.
46. Matsuzaki, M., Ellis-Davies, G.C.R., Nemoto, T.,
Miyashita, Y., Iino, M. and Kasai, H. Dendritic spine
geometry is critical for AMPA receptor expression in
hippocampal CA1 pyramidal neurons. Nature
Neurosci. 4, 1086-1092, 2001.
47. Yamazawa, T. and Iino, M. Simultaneous imaging of
Ca2+ signals in interstitial cells of Cajal and longitudinal smooth muscle cells during rhythmic activity inmouse ileum. J. Physiol. 538, 823 –835, 2002.
48. Oyamada, H., Oguchi, K., Saitoh, N., Yamazawa, T.,
Hirose, K., Kawana, Y., Wakatsuki, K., Oguchi, K.,
Tagami, M., Hanaoka, K., Endo, M. and Iino, M.
Novel mutations in C-terminal channel region of theryanodine receptor in malignant hyperthermia
patients. Jpn. J. Pharmacol. 88, 159-166, 2002.
49. Tu, H., Miyakawa, T., Wang, Z., Glouchankova, L.,
Iino, M. and Bezprozvanny, I. Functional characterization of the type 1 Inositol 1,4,5-trisphosphate
receptor coupling domain SII({+/-}) splice variants
and the opisthotonos mutant form. Biophys. J. 82,
1995-2004, 2002.
50. Ueda, H.R., Chen, W., Adachi, A., Wakamatsu, H.,
Hayashi, S., Takasugi, T., Nagano, M., Nakahama,
K.-I., Suzuki, Y., Sugano, S., Iino, M., Shigeyoshi, Y.
and Hashimoto, S. A transcription factor response
element for gene expression during circadian night.
Nature, 418, 534 – 539, 2002.

49

Department of
Molecular Neurobiology
Outline and Research Objective
We have been investigating the molecular mechanism of brain functions. Current
research activities are focused on the glutamate receptor (GluR) and learning and memory.
We elucidated the molecular diversity of the N-methyl-D-aspartate (NMDA)-type GluR anddiscovered the novel δ subfamily of GluR by molecular cloning. Roles of these GluRs inbrain functions have been studied by gene targeting.

Faculties and Students
Professor and Chair Masayoshi Mishina, Ph.D (1994~)
Lecturer
Hisashi Mori, Ph.D
Associate .................................2 (Naoto Matsuda, M.D andTomonori Takeuchi, Ph.D)
Postdoctoral Fellow ................2
Graduate Student..................11
Research Student ....................2
Secretary .................................2

Past Research and Major Accomplishments
The NMDA subtype of GluR is unique in functional properties among many neurotransmitter receptors
and ion channels mediating neural signaling in thebrain. The NMDA receptor channel is gated both byligands and by voltage, and is highly permeable toCa2+. These characteristics of the NMDA receptor
directly relate to its important physiological roles insynaptic plasticity as a molecular coincidence detector. Some forms of long-term potentiation (LTP) andlong-term depression (LTD), which are thought tounderlie learning and memory, are critically dependent on the NMDA receptor channel.

Molecular characterization of the NMDA receptor
We elucidated the molecular diversity of the
NMDA receptor by molecular cloning. Highly active
NMDA receptor channel was formed in vitro by coexpression of two members of GluR subunit families,that is, the GluRε (NR2) and GluRζ (NR1). There are
four members in the ε subunit family, whereas only
one member is known in the GluRζ subunit family
except for the splice variants. All of the NMDA receptor channel subunits possess asparagine in segment
M2. Replacement by glutamine of the asparagine insegment M2 of the GluRε2 and GluRζ1 strongly
reduced the sensitivity to Mg2+ block of the NMDA
receptor channel. Since there is strong evidence that
Mg2+ produces a voltage-dependent block of the channel by binding a site deep within the ionophore, these50

results are consistent with the view that segment M2
constitutes the ion channel pore of the NMDA receptor channel.
At the embryonic stages, the GluRε2 (NR2B) subunit mRNA is expressed in the entire brain, and the
GluRε4 (NR2D) subunit mRNA in the diencephalon
and the brainstem. After birth, the GluRε1 (NR2A)
subunit mRNA appears in the entire brain, and the
GluRε3 (NR2C) subunit mRNA mainly in the cerebellum. The expression of the GluRε2 subunit mRNA
becomes restricted to the forebrain, and that of the
GluRε4 subunit mRNA is strongly reduced. The
GluRζ1 subunit mRNA is found ubiquitously in thebrain during development. The four GluRε subunits
are also distinct in functional properties and regulation. Thus, multiple GluRε subunits are major determinants of the NMDA receptor channel diversity, andthe molecular compositions and functional properties
of NMDA receptor channels are different dependingon the brain regions and developmental stages. Thesefindings raise an important question whether themolecular diversity underlies the various physiologicalroles of the NMDA receptor channel.

Physiological roles of multiple NMDA receptor
subtypes
To examine the functional roles in vivo of thediverse NMDA receptor subtypes, we generated
mutant mice defective in respective GluRε subunits bygene targeting. Disruption of the GluRε1 gene results
in reduction of hippocampal LTP and impairment of
Morris water maze learning. In GluRε1 mutant mice,
thresholds for both hippocampal LTP and contextual
learning increased. The ablation of the GluRε2 subunit also impaired synaptic plasticity in the hippocampus. The reduction of GluRε1 and GluRε2 affected theplasticity of the hippocampal CA3 region in a synapsespecific manner. These results indicate that the
NMDA receptor GluRε1 and GluRε2 subtypes play a
key role in synaptic plasticity, learning and memory.
GluRε2 mutant mice died shortly after birth andfailed to form the whisker-related neural pattern (bar-relettes) in the brainstem trigeminal complex. In contrast, the barrelette formation was normal in GluRε4mutant mice. These results show the involvement of
the GluRε2 subunit in the refinement of the synapse
formation of periphery-related neural patterns in the
mammalian brain. Heterozygous mutant mice with
reduced GluRε2 subunit of the NMDA receptor
showed strongly enhanced startle responses toacoustic stimuli. On the other hand, heterozygous andhomozygous mutation of the other NMDA receptor
GluRε subunits exerted little or only small effects on
acoustic startle responses. Thus, the NMDA receptor
GluRε2 plays a role in the regulation of the startle
reflex. GluRε4 mutant mice exhibited reduced spontaneous activity, while GluRε3 mutant mice showed little obvious deficit. GluRε1 and GluRε4 differentiallycontributed to pain modulation.

Discovery and functional roles of GluRδ2 in the
cerebellum
The wealth of knowledge on the neural circuits inthe cerebellum makes the cerebellum an ideal system
to study the molecular mechanism of brain function.
By molecular cloning, we found a novel GluR subfamily named GluRδ. GluRδ2 was selectively localized in
cerebellar Purkinje cells. Furthermore, its intracellular
localization was restricted to the parallel fiberPurkinje cell synapses. The carboxyl terminus of
GluRδ2 interacted with delphilin containing a single
PDZ domain, formin homology (FH) domains and acoiled-coil structure. Analyses of GluRδ2 mutant mice
revealed that the GluRδ2 subunit was essential inmotor coordination and cerebellar LTD and in refinement and maintenance of Purkinje cell synapses.
We investigated eyeblink conditioning in GluRδ2mutant mice to elucidate its cerebellar cortical neuralmechanism, with reference to the temporal relationship of conditioned and unconditioned stimuli. In the
delay paradigm, in which a tone (CS) overlapped temporally with a periorbital shock (US), GluRδ2 mutant
mice exhibited a severe impairment in learning.
However, in the trace paradigm in which a stimulusfree trace interval up to 500 ms intervened between
the CS and US, GluRδ2 mutant mice learned as successfully as the wild-type mice even with 0 ms-traceinterval. We then examined the delay and trace eyeblink conditioning in NMDA receptor GluRε1 mutant
mice. In delay conditioning, GluRε1 mutant miceattained a normal level of the conditioned response(CR), although acquisition was a little slower than inwild-type mice. In contrast, GluRε1 mutant mice
exhibited severe impairment of the attained level of
the CR and disturbed temporal pattern of CR expression in trace conditioning with a longer trace intervalof 500 ms. These results suggest that neural substrates underlying eyeblink conditioning are different

depending on the temporal overlap of the conditioned
and unconditioned stimuli.

Current Research
These studies show that GluRs play key roles inmemory acquisition and neural pattern formation.
The memory signaling in the adult brain may sharethe common molecular mechanism with the activitydependent synapse refinement during neural development. To further investigate the molecular basis ofhigher brain function, we are developing the conditional gene targeting in the C57BL/6 mouse genetic
background and the molecular genetics of zebrafish.

Conditional gene targeting in C57BL/6 genetic
backgroundWe developed an efficient homologous recombination system using ES cells derived from C57BL/6
strain. For a cell type-specific and temporal regulationof gene targeting in the brain, we have generated
mouse lines that express Cre recombinase-progesterone receptor fusion (CrePR) gene specifically cerebellar granule cells, cerebellar Purkinje cells, hippocampal CA3 pyramidal neurons and striatal spiny
neurons. Crossing of target mice first with FLP mice
to remove the neo selection marker gene and thenwith Cre mice has yielded mutant mice lacking GluR
genes in specific neurons.
Molecular genetics of zebrafish
Elucidation of how the neural network is formed
and modulated is essential to understand how thebrain is functioning. The retinotectal projection andolfactory systems in transparent zebrafish are suitable
to analyze synapse formation in vivo. We are developing a novel strategy that visualizes and manipulates
developing neurons in vivo.

Future Prospects
Combination of molecular genetic approaches inmice and zebrafish will facilitate our understanding of
the mechanism of higher brain function at the molecular, cellular and neural network levels.

Research Grants
1. Grant-in-Aid for Scientific Research on Priority
Areas, The Ministry of Education, Culture, Sports,
Science and Technology (1995-1999)
2. Grant-in-Aid for Scientific Research (A), The
Ministry of Education, Culture, Sports, Science andTechnology (1996-1998)
3. CREST, Japan Science and Technology Corporation(1996-2001)

51

4. Grant-in-Aid for Scientific Research on Priority
Areas, The Ministry of Education, Culture, Sports,
Science and Technology (2000-)
5. SORST, Japan Science and Technology Corporation(2002-)

Select Publications
1. Mishina, M., Kurosaki, T., Tobimatsu, T., Morimoto,Y., Noda, M., Yamamoto, T., Terao, M., Lindstrom,J., Takahashi, T., Kuno, M. and Numa, S. Expressionof functional acetylcholine receptor from cloned
cDNAs. Nature 307, 604-608, 1984.
2. Mishina, M., Tobimatsu, T., Imoto, K., Tanaka, K.,
Fujita, Y., Fukuda, K., Kurasaki, M., Takahashi, H.,
Morimoto, Y., Hirose, T., Inayama, S., Takahashi, T.,
Kuno, M. and Numa, S. Location of functional
regions of acetylcholine receptor α-subunit by sitedirected mutagenesis. Nature 313, 364-369, 1985.
3. Takai, T., Noda, M., Mishina, M., Shimizu, S.,
Furutani, Y., Kayano, T., Ikeda, T., Kubo, T.,
Takahashi, H., Takahashi, T., Kuno, M. and Numa,
S. Cloning, sequencing and expression of cDNA for anovel subunit of acetylcholine receptor from calfmuscle. Nature 315, 761-764, 1985.
4. Sakmann, B., Methfessel, C., Mishina, M.,
Takahashi, T., Takai, T., Kurasaki, M., Fukuda, K.
and Numa, S. Role of acetylcholine receptor subunits
in gating of the channel. Nature 318, 538-543, 1985.
5. Mishina, M., Takai, T., Imoto, K., Noda, M.,
Takahashi, T., Numa, S., Methfessel, C. andSakmann, B. Molecular distinction between fetal andadult forms of muscle acetylcholine receptor. Nature
321, 406-411, 1986.
6. Kubo, T., Fukuda, K., Mikami, A., Maeda, A.,
Takahashi, H., Mishina, M., Haga, T., Haga, K.,
Ichiyama, A., Kangawa, K., Kojima, M., Matsuo, H.,
Hirose, T. and Numa, S. Cloning, sequencing andexpression of complementary DNA encoding themuscarinic acetylcholine receptor. Nature 323, 411416, 1986.
7. Imoto, K., Methfessel, C., Sakmann, B., Mishina, M.,
Mori, Y., Konno, T., Fukuda, K., Kurasaki, M., Bujo,
H., Fujita, Y. and Numa, S. Location of a δ-subunit
region determining ion transport through the acetylcholine receptor channel. Nature 324, 670-674, 1986.
8. Fukuda, K., Kubo, T., Akiba, I., Maeda, A., Mishina,
M. and Numa, S. Molecular distinction between
muscarinic acetylcholine receptor subtypes. Nature
327, 623-625, 1987.
9. Fukuda, K., Higashida, H., Kubo, T., Maeda, A.,
Akiba, I., Bujo, H., Mishina, M. and Numa, S.
Selective coupling with K+ currents of muscarinic
acetylcholine receptor subtypes in NG108-15 cells.
Nature 335, 355-358, 1988.
10. Imoto, K., Busch, C., Sakmann, B., Mishina, M.,
Konno, T., Nakai, J., Bujo, H., Mori, Y., Fukuda, K.
and Numa, S. Rings of negatively-charged aminoacids determine the acetylcholine receptor channel
conductance. Nature 335, 645-648, 1988.

52

11. Meguro, H., Mori, H., Araki, K., Kushiya, E.,
Kutsuwada, T., Yamazaki, M., Kumanishi, T.,
Arakawa, M., Sakimura, K. and Mishina, M.
Functional characterization of a heteromeric NMDA
receptor channel expressed from cloned cDNAs.
Nature 357, 70-74, 1992.
12. Kutsuwada, T., Kashiwabuchi, N., Mori, H.,
Sakimura, K., Kushiya, E., Araki, K., Meguro, H.,
Masaki, H., Kumanishi, T., Arakawa, M. andMishina, M. Molecular diversity of the NMDA receptor channel. Nature 358, 36-41, 1992.
13. Mori, H., Masaki, H., Yamakura, T. and Mishina, M.
Identification by mutagenesis of a Mg2+ block site of
the NMDA receptor channel. Nature 358, 673-675,
1992.
14. akimura, K., Kutsuwada, T., Ito, I., Manabe, T.,
Takayama, C., Kushiya, E., Yagi, T., Aizawa, S.,
Inoue, Y., Sugiyama, H. and Mishina, M. Reduced
hippocampal LTP and spatial learning in mice lacking NMDA receptor ε1 subunit. Nature 373, 151-155,
1995.
15. Kashiwabuchi, N., Ikeda, K., Araki, K., Hirano, T.,
Shibuki, K., Takayama, C., Inoue, Y., Kutsuwada, T.,
Yagi, T., Kang, Y., Aizawa, S. and Mishina, M.
Impairment of motor coordination, Purkinje cell
synapse formation and cerebellar long-term depression in GluRδ2 mutant mice. Cell 81, 245-252, 1995.
16. Kutsuwada, T., Sakimura, K., Manabe, T., Takayama,C., Katakura, N., Kushiya, E., Natsume, R.,
Watanabe, M., Inoue, Y., Yagi, T., Aizawa, S.,
Arakawa, M., Takahashi, T., Nakamura, Y., Mori, H.
and Mishina, M. Impairment of suckling response,
trigeminal neuronal pattern formation and hippocampal LTD in NMDA receptor ε2 subunit mutant
mice. Neuron 16, 333-344, 1996.
17. Takahashi, T., Feldmeyer, D., Suzuki, N., Onodera,
K., Cull-Candy, S. G., Sakimura, K. and Mishina, M.
Functional correlation of NMDA receptor ε subunits
expression with the properties of single-channel andsynaptic currents in the developing cerebellum. J.
Neurosci. 16, 4376-4382, 1996.
18. Ito, I., Futai, K., Katagiri, H., Watanabe, M.,
Sakimura, K., Mishina, M. and Sugiyama, H.
Synapse-selective impairment of NMDA receptor
functions in mice lacking NMDA receptor ε1 or ε2subunit. J. Physiol. 500, 401-408, 1997.
19. Taniguchi, M., Yuasa, S., Fujisawa, H., Naruse, I.,
Saga, S., Mishina, M. and Yagi, T. Disruption of semaphorin III/D gene causes severe abnormality inperipheral nerve projection. Neuron 19, 519-530,
1997.
20. Kurihara, H., Hashimoto, K., Kano, M., Takayama,C., Sakimura, K., Mishina, M., Inoue, Y. and
Watanabe, M. Impaired parallel fiber-Purkinje cell
synapse stabilization during cerebellar developmentof mutant mice lacking the glutamate receptor δ2
subunit. J. Neurosci. 17, 9613-9623, 1997.
21. Watanabe, M., Fukaya, M., Sakimura, K., Manabe,T., Mishina, M. and Inoue, Y. Selective scarcity of
NMDA receptor channel subunits in the stratum

lucidum (mossy fiber-recipient layer) of the hippocampal CA3 subfield. Eur. J. Neurosci. 10, 478-487,
1998.
22. Ando, H. and Mishina, M. Efficient mutagenesis ofzebrafish by a DNA cross-linking agent. Neurosci.
Lett. 244, 81-84, 1998.
23. Kohmura, N., Senzaki, K., Hamada, S., Kai, N.,
Yasuda, R., Watanabe, M., Ishii, H., Yasuda, M.,
Mishina, M. and Yagi, K. Diversity revealed by anovel family of cadherins expressed in neurons at
synaptic complex. Neuron 20, 1137-1151, 1998.
24. Kiyama, Y., Manabe, T., Sakimura, K., Kawakami, F.,
Mori, H. and Mishina, M. Increased thresholds forlong-term potentiation and contextual learning inmice lacking the NMDA-type glutamate receptor ε1
subunit. J. Neurosci. 18, 6704-6712, 1998.
25. Mori, H., Manabe, T., Watanabe, M., Satoh, Y.,
Suzuki, N., Toki, S., Nakamura, K., Yagi, T., Kushiya,E., Takahashi, T., Inoue, Y., Sakimura, K. andMishina, M. Role of the carboxyl-terminal region of
the GluRε2 subunit in synaptic localization of the
NMDA receptor channel. Neuron 21, 571-580, 1998.
26. Morikawa, E., Mori, H., Kiyama, Y., Mishina, M.,
Asano, T. and Kirino, T. Attenuation of focal
ischemic brain injury in mice deficient in the ε1(NR2A) subunit of NMDA receptor. J. Neurosci. 18,
9727-9732, 1998.
27. Hayashi, T., Umemori, H., Mishina, M. andYamamoto, T. The AMPA receptor interacts withand signals through the protein tyrosine kinase Lyn.
Nature 397, 72-76, 1999.
28. Furuyashiki, T., Fujisawa, K., Fujita, A., Madaule, P.,
Uchino, S., Mishina, M., Bito, H. and Narumiya, S.
Citron, a Rho-target, interacts with PSD-95/SAP-90 at
glutamatergic synapses in the thalamus. J. Neurosci.
19, 109-118, 1999.
29. Tsujita, M., Mori, H., Watanabe, M., Suzuki, M.,
Miyazaki, J. and Mishina, M. Cerebellar granule cellspecific and inducible expression of Cre recombinase
in the mouse. J. Neurosci. 19, 10318-10323, 1999.
30. Minami, T., Okuda-Ashitaka, E., Mori, H., Sakimura,K., Watanabe, M., Mishina, M. and Ito, S.
Characterization of nociceptin/orphanin FG-induced
pain responses in conscious mice: Neonatal capsaicin
treatment and N-methyl-D-aspartate receptor GluRεsubunit knockout mice. Neuroscience 97, 133-142,
2000.
31. Hironaka, K., Umemori, H., Tezuka, T., Mishina, M.
and Yamamoto, T. The protein-tyrosine phosphatase
PTPMEG interacts with glutamate receptor δ2 and εsubunits. J. Biol. Chem. 275, 16167-16173, 2000.
32. Matsuda, I. and Mishina, M. Identification of a juxtamembrane segment of the glutamate receptor δ2
subunit required for the plasma membrane localization. Biochem. Biophys. Res. Commun. 275, 565-571,
2000.
33. Nakamura, K., Manabe, T., Watanabe, M., Mamiya,T., Ichikawa, R., Kiyama, Y., Sanbo, M., Yagi, T.,
Inoue, Y., Nabeshima, T., Mori, H. and Mishina M.
Enhancement of hippocampal LTP, reference memo-

ry and sensorimotor gating in mutant mice lacking atelencephalon-specific cell adhesion molecule. Eur. J.
Neurosci. 13, 179-189, 2001.
34. Nakazawa, T., Komai, S., Tezuka, T., Hisatsune, C.,
Umemori, H., Semba, K., Mishina, M., Manabe, T.
and Yamamoto, T. Characterization of Fyn-mediatedtyrosine phosphorylation sites on GluRε2 (NR2B)
subunit of the N-methyl-D-aspartate receptor. J. Biol.
Chem. 276, 693-699, 2001.
35. Miyamoto, Y., Yamada, K., Noda, Y., Mori, H.,
Mishina, M. and Nabeshima, T. Hyperfunction of
dopaminergic and serotonergic neural systems inmice lacking NMDA receptor ε1 subunit. J. Neurosci.
21, 750-757, 2001.
36. Uchino, S., Nakamura, T., Nakamura, K., NakajimaIijima, S., Mishina, M., Kohsaka, S. and Kudo, Y.
Real-time two-dimensional visualization of ischemiainduced glutamate release from hippocampal slices.
Eur. J. Neurosci. 13, 670-678, 2001.
37. Kishimoto, Y., Kawahara, S., Mori, H., Mishina, M.
and Kirino, Y. Long-trace interval eyeblink conditioning is impaired in mutant mice lacking the NMDA
receptor subunit ε1. Eur. J. Neurosci. 13, 1221-1227,
2001.
38. Kishimoto, Y., Kawahara, S., Suzuki, M., Mori, H.,
Mishina, M. and Kirino, Y. Classical eyeblink conditioning in glutamate receptor subunit δ2 mutant
mice is impaired in the delay paradigm but not in the
trace paradigm. Eur. J. Neurosci. 13, 1249-1253,
2001.
39. Kitayama, K., Abe, M., Kakizaki, T., Honma, D.,
Natsume, R., Fukaya, M., Watanabe, M., Miyazaki,J., Mishina, M. and Sakimura, K. Purkinje cell-specific and inducible gene recombination system generated from C57BL/6 mouse ES cells. Biochem. Biophys.
Res. Commun. 281, 1134-1140, 2001.
40. Takeuchi, T., Kiyama, Y., Nakamura, K., Tsujita, M.,
Matsuda, I., Mori, H., Munemoto, Y., Kuriyama, H.,
Natsume, R., Sakimura, K. and Mishina, M. Roles of
the glutamate receptor ε2 and δ2 subunits in thepotentiation and prepulse inhibition of the acoustic
startle reflex. Eur. J. Neurosci. 14, 153-160, 2001.
41. Wainai, T., Takeuchi, T., Seo, N. and Mishina, M.
Regulation of acute nociceptive responses by the
NMDA receptor GluRε2 subunit. Neuroreport 12,
3165-3172, 2001.
42. Kishimoto, Y., Kawahara, S., Fujimichi, R., Mori, H.,
Mishina M., and Kirino Y. Impairment of eyeblinkconditioning in GluRδ2 mutant mice depends on thetemporal overlap between conditioned and unconditioned stimuli. Eur. J. Neurosci. 14, 1515-1521, 2001.
43. Hashimoto, K., Ichikawa, R., Takechi, H., Inoue, Y.,
Aiba, A., Sakimura, K., Mishina, M., Hashikawa, T.,
Konnerth, A., Watanabe, M., and Kano, M. Roles of
GluRδ2 and mGluR1 in climbing fiber synapse elimination during postnatal cerebellar development. J.
Neurosci. 21, 9701-9712, 2001.
44. Takatsuki, K., Kawahara, S., Mori, H., Mishina, M.
and Kirino, Y. Scopolamine impairs eyeblink condi-

53

tioning in the cerebellar LTD-deficient mice.
Neuroreport 13, 159-162, 2002.
45. Miyagi, Y., Yamashita, T., Fukaya, M., Sonoda, T.,Okuno, T., Yamada, K., Watanabe, M., Nagashima,Y., Aoki, I., Okuda, K., Mishina, M. and Kawamoto,S. Delphilin: a novel PDZ- and formin homologydomain containing protein, which synaptically colocalizes and interacts with glutamate receptor δ2 subunit. J. Neurosci. 22, 803-814, 2002.
46. Miyamoto, Y., Yamada, K., Noda, Y., Mori, H.,
Mishina, M. and Nabeshima, T. Lower sensitivity tostress and altered monoaminergic neuronal function
in mice lacking the NMDA receptor ε4 subunit. J.
Neurosci. 22, 2335-2342, 2002.
47. Takeuchi, T., Nomura, T., Tsujita, M., Suzuki, M.,
Fuse, T., Mori, H. and Mishina, M. Flp recombinasetransgenic mice of C57BL/6 strain for conditional
gene targeting. Biochem. Biophys. Res. Commun.
293, 953-957, 2002.
48. Yoshida, T., Ito, A., Matsuda, N. and Mishina, M.
Regulation by protein kinase A switching of axonal
pathfinding of zebrafish olfactory sensory neurons
through the olfactory placode-olfactory bulb boundary. J. Neurosci. 22, 4964-4972, 2002.
49. Uemura, T., Mori, H. and Mishina, M. Isolation andcharacterization of Golgi apparatus-specific GODZ
with the DHHC zinc finger domain. Biochem.
Biophys. Res. Commun. 296, 492-496, 2002.
50. Ichikawa, R., Miyazaki, T., Kano, M., Hashikawa, T.,
Tatsumi, H., Sakimura, K., Mishina, M., Inoue, Y.,
and Watanabe, M. Distal extension of climbing fiber
territory and multiple innervation caused by aberrant
wiring to adjacent spiny branchlets in cerebellar
Purkinje cells lacking glutamate receptor GluRδ2. J.
Neurosci. 22, 8487-8503, 2002.
51. Tokuoka, H., Yoshida, T., Matsuda, N. and Mishina,
M. (2002) Regulation by GSK-3β of the arborization
field and maturation of retinotectal projection inzebrafish. J. Neurosci., in press.

54Department of
Human Pathology
Outline and Research Objectives
The Department of Human Pathology was established in 1997, originating from the First
Department of Pathology of the Faculty of Medicine. The Department of Human Pathology is
specifically dedicated to the research of anatomical and surgical pathology of human diseases.
The Department of Pathology was established in 1887 and has a long history of bothanatomical and experimental pathology. Professor Dr. Katsusaburo Yamagiwa (professor:
1905-1923) was the first in the world who succeeded in inducing skin carcinoma on rabbit
ears with coal tar in 1908. He was also known to publish a book regarding the carcinogenesis of the stomach. The First and the Second Department Pathology, like as one unit, were
engaged in research, diagnosis and teaching. The main research topics of the Department in
one hundred years, either anatomical or experimental, varied according to the specialties of
its professors and the academic demands of the time in Japan. A brief list of the topics
includes clonorchiasis, tsutsugamushi disease, beriberi and salivary gland hormones whichwere investigated before World War II, and thereafter pathology of atomic bomb injuries,
tuberculosis, hepatic disorders, bone marrow pathology and neoplastic diseases of variousorgans (e.g., stomach, liver, and bone). The experimental aspects of pathology are now succeeded and have been developed by the Department of Molecular Pathology.
The practice of anatomical pathology was a basis of research in the past, but it still constitutes a source of ideas even in the research-oriented pathology. The number of autopsies conducted from 1883 to the present has reached 33,950 and in spite of the current decline in the
autopsy rate, one hundred autopsies are still performed each year. Surgical pathology has
increasingly become important in the practice of pathology. The laboratory of surgical pathology in the University Hospital, which was established in 1955, became an independent division of the central laboratory of the hospital in 1975. After the integration of the Branch
Hospital into the University Hospital in 2001, the numbers of histological and cytological
specimens examined each year are expected to be more than 12,000 and 20,000, respectively.
The division has been run with the cooperation of the Department of Pathology.
The Department of Human Pathology has succeeded the classical aspects of the
Department of Pathology. Its mission is to be the bridge between clinical medicine and basic
life sciences. Its research objectives are (1) to discover a new entity of human diseases basedon the practice of anatomical and surgical pathology, (2) to clarify disease mechanisms basedon morphology, and (3) to function as a “museum” for various forms of human disorders.

Faculties and Students
Professor and Chair

Masashi Fukayama, M.D., D.M.Sc.
(1999-)
Associate Professors Toshiro Niki, M.D., D.M.Sc., Ph.D.
Lecturer
Ja-Mun Chong, M.D., D.M.Sc.
Associate .................................3
Graduate student ..................13
Research student.....................2

Past Research and Major Accomplishments
Our research projects in the past ten years attempt
to investigate the role of chronic inflammation in thedevelopment and progression of carcinoma. We
focused our investigations on the following diseases:(1) Epstein-Barr virus (EBV)-associated neoplasms,
such as gastric carcinomas, pyothorax-associated lymphoma (PAL), Hodgkin’s disease and AIDS lymphomas, and (2) lung carcinomas in patients with pulmonary fibrosis and lung adenocarcinoma with central scar formation.
1. EBV-associated neoplasms
EBV is the first virus that was identified in a humanneoplastic cell in 1963. More than 90% of the world population is infected with EBV before adolescence, and it isthought that certain small populations develop EBVassociated malignancy in an endemic manner, such as
Burkitt’s lymphoma in equatorial Africa and nasopharyngeal carcinoma in Southern China. However, recent
advances in molecular biological techniques have55

demonstrated that various neoplasms in the general
population are associated with EBV infection.
1-1) EBV-associated gastric carcinoma
EBV-associated gastric carcinoma is now considered to be the most common among EBV-associatedneoplasms, and its frequency is 10% or less of gastriccarcinomas in Japan. We found that EBV-associatedgastric carcinoma is a distinct subgroup of gastric carcinoma, and that it develops against a background ofsevere atrophic gastritis. In our evaluation of genetic
abnormalities, EBV-associated gastric carcinoma wasfound to be remarkable in its paucity of loss of heterozygosity (LOH) and microsatellite instability.
Since it is difficult to maintain epithelial cells in
vitro in the latent form of EBV infection, we established one strain of EBV-associated gastric carcinoma
transplantable to severe combined immunodeficiency(SCID) mice, which faithfully maintains the original
EBV and the expression of EBV-related genes.
Applying the DNA-chip analysis to the gastric carcinoma strains with or without EBV-infection in SCID
mice, we clarified the expression profile specific to
EBV-associated gastric carcinoma, and found that theexpression of IL1-β is markedly up-regulated in thistype of carcinoma. Of interest is the observation that
IL1- β facilitates the growth of a gastric cancer cellline, suggesting that it is an autocrine growth factor in
EBV-associated gastric carcinoma.
1-2) EBV-associated lymphomas
PAL develops within the pleural cavity of patients
who had a 20-50-year history of pyothorax following
tuberculous pleuritis or an artificial pneumothorax forthe treatment of pulmonary tuberculosis. Although
the reason is not clear at present, most patients of
PAL are Japanese. We discovered in 1993 that EBV iscausally associated with PAL and that PAL shows thesame expression pattern of EBV latency genes as thatof AIDS lymphomas.
2. Lung carcinoma
Lung cancer is the major cause of cancer-related
deaths among Japanese. Since one of our specialties isthe pathology of lung and thymus, two issues are underinvestigation, that is, the relationship between pulmonary fibrosis and development of lung cancer, and therole of fibrosis in the progression of adenocarcinoma.
2-1) Idiopathic pulmonary fibrosis and lung carcinoma
We evaluated the frequency of lung cancer among
the patients with idiopathic pulmonary fibrosis (IPF)by performing autopsy-based case control study.
Then, we morphologically evaluated the atypicalepithelial proliferation in the honeycombed area.

56

Although squamous metaplasia is much more frequent in the honeycombed area of IPF patients withlung carcinoma, we confirmed the necessity to evaluate the genetic abnormalities in minute foci of atypicalepithelial cells in tissue sections.
2-2) Small adenocarcinoma
Among nonsmall cell carcinomas of the lung, theprognosis of adenocarcinoma is relatively poor. Onethird of the patients with stage I adenocarcinoma dieof the disease within five years after surgery, and the
5-year recurrence rate is as high as 20% for adenocarcinomas of less than 2 cm in diameter. In our study ofsmall adenocarcinoma, inactivation of p16 may play arole in accelerating scar formation and lymph nodemetastasis, and through these mechanisms p16-inactivation may contribute to a poor prognosis of thepatients.

Current Research
1. EBV-associated neoplasms
1-1) Global methylaion in EBV-associated gastriccarcinoma
Recently we discovered the phenomenon of global
and nonrandom methylation of the promoter regionof cancer-associated genes in EBV-associated gastriccarcinoma. Such a global methylation was notobserved in EBV-negative carcinomas. One of theimportant roles of DNA methylation is to act as adefense against extrinsic pathogens. The latent infection of viruses in either episomal (EBV), or integrated
form (human immunodeficiency virus type 1 orhuman T-cell leukemia-virus type-1), is restricted bymethylation of viral genomes. Thus, we are now
investigating a possible mechanism; that is, overridden methylation of cellular genes suppresses exprssion of important genes of EBV-infected epithelial
cells, leading to the development of carcinoma.
1-2) EBV-associated lymphomas and human herpes virus 8
Human herpes virus 8 (HHV8) was first discoveredin tissues of Kaposi sarcoma in 1994, and is associatedwith primary effusion lymphoma and multicentric
Castleman’s disease. However, its epidemiology inlymphoproliferative disorders and its relation to EBV
have not been fully clarified. We are now investigating the molecular epidemiology of HHV8-associateddiseases in Japan and Asian countries.

2. Lung carcinoma
2-1) Laser capture microscopy-assisted analysis ofprecursor lesions
Laser capture microscopy (LCM) is now widely
applied to analysis of precursor lesions of various carcinomas. We are also applying it to the evaluation ofepithelial cells of honeycombed areas in the diseased
lungs of IPF patients, and atypical epithelial cells of
adenomatous hyperplasia, a precursor of bronchioloalveolar carcinoma of the lung. Nuclei of target
cells directly obtained from paraffin-embedded sections are subjected to PCR-based analysis.
Invasion-associated gene expression: Niki et al
demonstrated the colocalization of cox-2 and laminin5 at the invasive front of early-stage lung adenocarcinomas. Current data suggest that p53 abnormalities
and EGFR signaling are involved in the aberrant
expression of these proteins. Coregulation and interaction of these molecules at the invasive front of cancer may facilitate tumor angiogenesis and invasion ina coordinated manner. Several on-going projects
investigate the expression of invasion-associated proteins in lung adenocarcinoma, including S100-related
proteins. We are also developing a technique thatenables us to apply DNA-chip analysis to tiny tissuesof the invasive front using the LCM-assisted sampling.

years, a new molecular approach will clarify changesof disease profiles in Japan in the 20th century.

2-2) Morphological analysis of lung carcinoma andthymic tumors
It is still necessary to review the histology of theseneoplasms in order to develop an evaluation system
for invasiveness and to identify the specific subgroupsof neoplasms.

1. Fukayama M, Hayashi Y, Takizawa t, Koike M, Endo
S and Okumura H. Glycoprotein hormone alpha-subunit in human stomach. J Histochem Cytochem 34,
995-1001, 1986.
2. Fukayama M, Hayashi Y, Koike M, Hajikano H and
Okumura H. Human chorionic gonadotropin in lungand lung tumors. Immunohistochemical study on
unbalanced distribution of subunits. Lab Invest 55,
433-443, 1986.
3. Fukayama M, Hayashi Y and Koike M. Human chorionic gonadotropin in gastric carcinoma. An immunohistochemical study suggesting independent regulation of subunits. Virchows Arch (Pathol Anat) 411,
205-212, 1987
4. Fukayama M and Koike M. So-called sclerosing
hemangioma of the lung. An immunohistochemical,
histochemical and ultrastructural study. Acta Pathol
Jpn 38, 627-642, 1988.
5. Fukayama M, Maeda Y, Funata N, Koike M, Saito K,
Sakai T and Ikeda T. Pulmonary and pleural thymoma. Diagnostic application of lymphocyte markers to
the thymoma of unusual site. Am J Clin Pathol 89,
617-621, 1988.
6. Fukayama M, Funata N, Hayashi Y, Maeda Y, Koike
M, Wantanabe J and Okabe T: Brain associatedsmall-cell lung cancer antigen (BASCA) in the lungand lung tumor. Immunohistochemical and immunoelectron microscopic study. J Histochem Cytochem
38, 51-57, 1990.

Future Prospects
There are two opposite directions that pathology is
expected to pursue at present and in the future. One isto be included among the fields of life science and cellbiology. The other is to remain a basic field for clinicalmedicine, which in its essence is a good partner andcritic of clinical medicine. To keep up with the growing demand for an intimate relationship with clinicalmedicine, we have decided to combine our department
with the Division of Surgical Pathology of the
University Hospital. The union is expected to facilitatecross talks between pathology and clinical medicine,which will lead us to our goal, namely, the discovery
of a new entity of human diseases and the clarificationof their mechanisms based on morphology.
We will also be obligated to maintain the museum
function of human pathology. Since we have archivesof paraffin blocks, including more than 20,000 autopsy specimens that were accumulated in these past 100

Research Grants
1. 1997-1999: Grant in Aid for Scientific Research (B)from the Ministry of Education, Science, Sports andCulture of Japan. “Pulmonary fibrosis and development of lung carcinoma. Analysis of dysplasic andneoplastic epithelial cells of pulmonary fibrosis.”
2. 1999-2002: Grant in Aid for Scientific Research (B)from the Ministry of Education, Science, Sports andCulture of Japan. “Molecular epidemiology of humanherpes virus 8-associated diseases in Japan and Asia.”
3. 2001: Grant in Aid for Scientific Research on a
Priority Area (C) from the Ministry of Education,
Science, Sports and Culture of Japan. “ Establishmentof gene diagnosis system for lung adenocarcinoma.”
4. 2002: Grant in Aid for Scientific Research on a
Priority Area (C) from the Ministry of Education,
Culture, Sports, Science and Technology of Japan. “Establishment of gene diagnosis system for lung adenocarcinoma.”
5. 2002-2004: Grant in Aid for Scientific Research (A)from the Ministry of Education, Culture, Sports,
Science and Technology of Japan. “Pathogenesis of
Epstein-Barr virus-associated neoplasms.”

Select Publications57

7. Fukayama M, Hayashi Y, Shiozawa Y, Furukawa E,
Funata N and Koike M. Human chorionicgonadotropinα-subunit in endocrine cells of fibrotic
and neoplastic lung. Its mode of localization and thesize profile of granules. Lab Invest 62, 444-451, 1990.
8. Hayashi Y, Watanabe J, Nakata K, Fukayama M and
Ikeda H. A novel diagnostic method of pneumocystis
carinii. In situ Hybridization of ribosomal ribonucleic
acid with biotinylated oligonucleotide probes. Lab
Invest 63, 576-580, 1990.
9. Nakata K, Akagawa KS, Fukayama M, Hayashi Y,
Kadokura M and Tokunaga T: Granulocytemacrophage colony-stimulating factor promotes theproliferation of human alveolar macrophages in vitro.
J Immunol 147, 1266-1272, 1991.
10. Fukayama M, Kanno T, Brody S, Kirby M and Crystal
RG. Respiratory tract gene transfer. Transplantationof genetically modified T-lymphocytes directly to therespiratory surface. J Biol Chem 266, 18339-1844,
1991
11. Rosenfeld MA, Siegfried W, Yoshimura K, Yoneyama
K, Fukayama M, Stier LE, Paakko PK, Gilardi P,
Stratford-Perricaudet LD, Perricaudet M, Jallat S,
Pavirani A, Lecocq J and Crystal RG. Adenovirusmediated transfer of a recombinant a1-antitrypsin
gene to the lung epithelium in vivo. Sience 252, 431252, 1991.
12. Rosenfeld MA, Yoshimura K, Trapnell BC, Yoneyama
K, Rosenthal ER, Dalemans W, Fukayama M, Bargon
J, Stier LE, Stratford-Perricaudet L, Perricaudet M,
Guggino WB, Pavirani A, Lecocq J and Crystal RG. In
vivo transfer of the human cystic fibrosis transmembrane conductance regulator gene to the airway
epithelium. Cell 68, 143-155, 1992.
13. Kikuchi-Yanoshita R, Konishi M, Ito S, Seki M,
Tanaka K, Maeda Y, Iino H, Fukayama M, Koike M,
Mori T, Sakuraba H, Fukunari H, Iwama T and
Miyaki M. Genetic changes of both p53 alleles associated with the conversion from colorectal adenoma toearly carcinoma in familial adenomatous polyposis
and non-familial adenomatous polyposis patients.
Cancer Res 52, 3965-3971, 1992.
14. Fukayama M, Ibuka T, Hayashi Y, Ooba T, Koike M
and Mizutani S. Epstein- Barr virus in pyothoraxassociated pleural lymphoma. Am J Pathol 143, 10441049, 1993.
15. Hayashi Y, Fukayama M, Koike M, Kaseda S, Ikeda T
and Yokoyama T. Cell-cycle analysis detecting
nuclear antigens: Comparison with BrdU-in vivolabeling and application to lung tumors. Acta Pathol
Jpn 43, 313-319, 1993.
16. Fukayama M, Hayashi Y, Iwasaki Y, Chong J, Ooba
T, Takizawa T, Koike M, Mizutani S, Miyaki M andHirai K. Epstein-Barr virus (EBV)-associated gastriccarcinoma and EBV-infection of the stomach. Lab
Invest 71, 73-81, 1994.
17. Ibuka T, Fukayama M, Hayashi Y, Funata N, Koike
M, Ikeda T and Mizutani S. Pyothorax-associatedpleural lymphoma: A case evolving from T-cell richlymphoid infiltration to overt B-cell lymphoma in

58

association with Epstein-Barr virus. Cancer 73, 738743, 1994.
18. Iino H, Fukayama M, Maeda Y, Koike M, Mori T,
Takahashi T, Kikuchi -Yanoshita R, Miyaki M,
Mizuno S and Watanabe S. Molecular genetics forclinical management of colorectal carcinoma. 17p,
18q, and 22q loss of heterozygosity and decreased
DCC expression are correlated with the metastatic
potential. Cancer 73,1324-1331, 1994.
19. Miyaki M, Konishi M, Kikuchi-Yanoshita R,
Enomoto M, Igari T, Tanaka K, Muraoka M,
Takahashi H, Amada Y, Fukayama M, Maeda M,
Iwama T, Mishima Y and Koike M. Characteristics ofsomatic mutation of the adenomatous polyposis coli
gene in colorectal tumors. Cancer Res 54, 3011-3020,
1994.
20. Hishima T, Fukayama M, Fujisawa M, Hayashi Y,
Arai K, Funata N, Koike M: CD5 expression inthymic carcinoma. Am J Pathol 145:268-1994.
21. Chong J, Fukayama M, Hayashi Y, Takizawa T, Koike
M, Konishi M, Kikuchi-Yanoshita R, Miyaki M:
Microsatellite instability in the progression of gastriccarcinoma. Cancer Res, 54:4595-4597, 1994.
22. Fukayama M, Hayashi Y, Ooba T, Funata N, Ibuka T,
Koike M, Hebisawa A, Kurasawa A, Fukayama M,
Nakahiro K and Kudoh S. Pyothorax-associated lymphoma: development of Epstein-Barr virus-associatedlymphoma within the inflammatory cavity. Pathol
Int 45, 825-831, 1995
23. Egawa N, Fukayama M, Kawaguchi K, Hishima T,
Hayashi Y, Funata N, Ibuka T, Koike M, Miyashita H
and Tajima T. Relapsing oral and colonic ulcers withmonoclonal T-cell infiltration. A low grade mucosal
T-lymphoproliferative disease of the digestive tract.
Cancer 75, 1728-1733, 1995.
24. Shimada-Hiratuka M, Fukayama M, Hayashi Y,
Ushijima T, Suzuki M, Hishima T, Funata N and
Koike M, Watanabe T. Primary gastric T-cell lymphoma with and without human T-lymphotropic
virus type 1. Cancer 80, 292-303, 1997.
25. Harada M, Suzuki M, Ikeda T, Kaneko T, Harada S
and Fukayama M. Clonality in nevocellular nevus
and malignant melanoma. An expression-based clonality analysis at the X-linked genes by polymerase
chain reaction. J Invest Dermatol 109, 656-660, 1997.
26. Chong J-M, Fukayama M, Hayashi Y, Funata N,
Takizawa T, Koike M, Kikuchi-Yanoshita R, Miyaki
M and Mizuno S. CD44 variants in gastric carcinoma
in association with or without Epstein-Barr virus. Int
J Cancer 74:450-454, 1997
27. Chong J, Fukayama M, Hayashi Y, Hishima T,
Funata N, Koike M, Matsuya S, Konishi M and
Miyaki M. Microsatellite Instability and loss of heterozygosity in gastric lymphoma. Lab Invest 77, 639645, 1997.
28. Hishima T, Fukayama M, Hayashi Y, Fujii T, Arai K,
Shiozawa Y, Funata N and Koike M: Neuroendocrine
differentiation in thymic epithelial tumors with special reference to thymic carcinoma and atypical thymoma. Hum Pathol 29, 330-338, 1998.

29. Iwasaki Y, Chong J-M, Hayashi Y, Ikeno R, Arai K,
Kitamura M, Koike M, Hirai K and Fukayama M:
Establishment and characterization of a human
Epstein-Barr virus-associated gastric carcinoma in
SCID mice. J Virol 72, 8321-8326, 1998.
30. Sakurai S, Fukayama M, Kaizaki Y, Saito K,
Kanazawa K, Kitamura M, Iwasaki Y, Hishima T,
Hayashi Y and Koike M: Telomerase Activity in
Gastrointestinal Stromal Tumor (GIST). Cancer 83,
2060-2066, 1998.
31. Fukayama M, Chong JM and Kaizaki Y. Epstein-Barrvirus and gastric carcinoma. Gastric Cancer 1, 104114, 1998.
32. Ishida T, Takashima R, Fukayama M, Hamada C,
Hippo Y, Fujii T, Moriyama S, Matsuba C, Nakahori
Y, Morita H, Yazaki Y, Kodama T, Nishimura S and
Aburatani H. New DNA polymorphisms of human
MMH/OGG1 gene: prevalence of one polymorphism
among lung- adenocarcinoma patients in Japanese.
Int J Cancer 80, 18-21, 1999.
33. Sakurai S, Fukasawa T, Chong JM and Fukayama M.
Embryonic Form of Smooth Muscle Myosin Heavy
Chain (SMemb/MHC-B) in Gastrointestinal Stromal
Tumor and Interstitial Cells of Cajal. Am J Pathol
154, 23-28, 1999.
34. Kawabuchi B, Moriyama S, Hironaka M, Fujii T,
Koike M, Moriyama H, Nishimura Y, Mizuno S and
Fukayama M. p16 inactivation in small sized lungadenocarcinoma: its association of poor prognosis.
Int J Cancer 84, 49-53, 1999.
35. Hayashi Y, Fukayama M, Funata N, Hishima T, Oba
T and Koike M. Polymerase chain reaction-screeningof immunoglobulin heavy chain and T-cell receptor g
gene rearrangements. A practical approach tomolecular DNA analysis of non-Hodgkin’s lymphoma in asurgical pathology laboratory. Pathol Int 49, 110-117,
1999.
36. Kaizaki Y, Sakurai S, Chong JM and Fukayama M:
Atrophic gatritis, Epstein-Barr virus infection, andEpstein-Barr virus-associated gastric carcinoma.
Gastric Cancer 2, 101-108, 1999.
37. Koshiishi N, Chong JM, Fukasawa T, Ikeno R,
Tanaka A, Kanazawa K, Ogura M, Ebuchi M,
Takizawa T, Funata N and Fukayama M.
Microsatellite instability and loss of heterozygosity
in primary and secondary proliferative lesions of theparathyroid gland. Lab Invest 79, 1051-1058, 1999.
38. Sakurai S, Fukasawa T, Chong JM, Tanaka A andFukayama M. C-kit gene abnormalities in gastrointestinal stromal tumors (tumors of interstitial cell of
Cajal). Jpn J Cancer Res 90, 1321-1328, 1999.
39. Hironaka M and Fukayama M. Pulmonary fibrosis
and lung carcinoma: A comparative study of metaplastic epithelia in honeycombed areas of usual interstitial pneumonia with or without lung carcinoma.
Pathol Int 49, 1060-1066, 1999.
40. Hishima T, Fukayama M, Hayashi Y, Fujii T, Ooba T,
Funata N and Koike M. CD70 in thymic carcinoma.
Am J Surg Pathol 25, 742-746, 2000.

41. Kurihara K, Shimizu S, Chong JM, Hishima T, Funata
N, Kashiwagi H, Nagai H, Miyaki M and Fukayama
M. Nuclear localization of immunorective b-cateninis specific to familial adenomatous polyposis in papillary thyroid carcinoma. Jpn J Cancer Res 91, 11001102, 2000.
42. Fukasawa T, Chong JM, Sakurai S, Koshiishi N,
Ikeno R, Tanaka A, Matsumoto Y, Hayashi Y, Koike
M and Fukayama M. Allelic loss of 14q, and 22q,
NF2 mutation, and genetic instability occur independently of c-kit mutation in gastrointestinal stromal tumor. Jpn J Cancer Res 91, 1241-1249, 2000.
43. Fukayama M, Chong JM and Uozaki H. Pathology
and molecular pathology of Epstein-Barr virus-associated gastric carcinoma. Curr Top Microbiol Immunol
258, 91-102, 2001.
44. Takeuchi K, Morishita Y, Fukayama M and Mori S.
Marked decrease in the Epstein-Barr virus positivity
rate in nodular sclerosis subtype Hodgkin’s diseasein Tokyo: trend between 1955 and 1999. Br J
Haematol 113, 429-431, 2001.
45. Sakurai S, Oguni S, Hironaka M, Fukayama M,
Morinaga S and Saito K. Mutations in c-kit geneexons 9 and 13 in gastrointestinal stromal tumors
among Japanese. Jpn J Cancer Res 92, 494-498, 2001.
46. Hippo Y, Taniguchi H, Tsutsumi S, Machida N,
Chong JM, Fukayama M, Kodama T and Aburatani
H. Global gene expression analysis of gastric cancer
by oligonucleotide microarrays. Cancer Res, 62, 233240, 2002.
47. Niki T, Kohno T, Iba S, Moriya Y, Takahashi Y, Saito
M, Maeshima A, YamadaT, Matsuno Y, Fukayama
M, Yokota J and Hirohashi S. Frequent CoLocalization of Cox-2 and Laminin-5 gamma2 Chainat the Invasive Front of Early-Stage Lung
Adenocarcinomas. Am J Pathol, 160, 1129-1141,
2002.
48. Chong JM, Sakuma K, Sudo M, Osawa T, Ohara E,
Uozaki H, Shibahara J, Kuroiwa K, Tominaga S,
Hippo Y, Aburatani H, Funata N and Fukayama M.
Interleukin-1beta expression in human gastric carcinoma with Epstein-Barr virus infection. J Virol 76,
6825-6831, 2002.
49. Kakiuchi C, Ishida T, Sato H, Katano H, Ishiko T,
Mukai H, Kogi M, Kasuga N, Takeuchi K, Yamane K,
Fukayama M and Mori S. Secretion of interleukin-6
and vascular endothelial growth factor by spindlecell sarcoma complicating Castleman’s disease (socalled ‘vascular neoplasia’). J Pathol 197, 264-271,
2002.
50. Osawa T, Chong J-M, Sudo M, Sakuma K, Uozaki H,
Shibahara J, Nagai H, Funata N and Fukayama M.
Reduced Expression and Promotor Mehtylation of
p16 Gene in Epstein-Barr Virus-associated Gastric
Carcinoma. Jpn J Cancer Res in press.

59Department of
Molecular Pathology
Outline and Research Objectives
Our department has a more than 100-year history from its establishment as the
Department of Pathology. The Second Department of Pathology has changed its name to
the Department of Molecular Pathology; after the retirement of Dr. Takatoshi Ishikawa, Dr.
Miyazono began to work as the Professor of the Department.
Our research is focused on molecular mechanisms of carcinogenesis, using molecular biological techniques. Transforming growth factor-b (TGF-β) is a potent growth inhibitor for
many cells, and perturbations of TGF-b signaling result in progression of various cancers.
We are interested in the signaling mechanisms of the TGF-β superfamily, including TGF-β
and bone morphogenetic proteins (BMPs), and studying how TGF-β signals regulate progression of various cancers, e.g. colorectal cancer.
We are also interested in the pathogenesis of vascular diseases. Signals induced by TGFβ and BMPs play important roles in the development and homeostasis of blood vessels.
Abnormalities in TGF-β/BMP signals result in genesis of certain vascular disorders. We haverecently started to investigate the mechanisms of growth and differentiation of murine
embryonic stem (ES) cells. Using an in vitro system, we are trying to regulate the differentiation of ES cells into smooth muscle cells (SMCs) and endothelial cells (ECs). The results
will be useful for the development of new strategies for treatment of cancers and vasculardiseases.

Faculty and Students

Past Research and Major Accomplishments

Professor and Chair

Members of the TGF-β superfamily bind to two
different types of serine/threonine kinase receptors,
and activate intracellular signaling molecules, theSmads. Smads move into the nucleus and regulatetranscription of target genes, including c-myc andcyclin-dependent kinase inhibitors. Signaling by
Smads is regulated by various molecules, including
inhibitory Smads (I-Smads) and transcriptional corepressors, c-Ski and SnoN (Figure 1).
We isolated cDNAs for TGF-β type I receptor(TbR-I, also called ALK-5) in 1993 [3]. In addition, we
have cloned five additional type I receptors, which
turned out to be the type I receptors for BMPs andKohei Miyazono, M.D., D.M.S.
(from 2000)
Associate Professors Keiji Miyazawa, Ph.D.
Associates ...............................2
Postdoctoral Fellow ................1
Research Technicians .............4
Ph.D. Students ........................2
Master Course Students .........3
Visiting Researcher .................1
Visiting Ph.D. Students ..........7
Secretaries ...............................2

Figure 1. Signaling by TGF-β through serine/threonine kinase receptors and Smads

60

activins [6], and the type II receptor for BMPs (BMPRII) [11]. Using these receptors, we demonstrated how
the members of the TGF-β superfamily bind to different combinations of type II and type I receptors.
During our studies of intracellular signaling activated by the serine/threonine kinase receptors, wehave cloned one of the mammalian I-Smads, Smad6[17], and reported that Smads can be classified into
three subclasses, i.e. R-Smads, Co-Smads, and I-Smads[18]. Moreover, we demonstrated how I-Smads inhibit
signaling activities of the TGF-β superfamily
cytokines [27, 29, 44]. We have shown that E3 ubiquitin ligases, Smurfs, interact with I-Smads, recruit
them from the nucleus to the cell surface receptor
complexes, and induce ubiquitination and degradation of the type I receptors [40, 48]. These findings
suggested that Smurfs support the inhibitory activityof I-Smads in TGF-β superfamily signaling.
We have also investigated the mechanisms bywhich Smads regulate transcription of target genes of
the TGF-β superfamily cytokines. We have shownthat Smads interact with various transcription factors,
including the vitamin D3 receptor and Runx proteins(also known as Cbfa/AML/PEBP2α proteins) [25, 30,37], which play important roles for TGF-β superfamily
cytokines in exhibiting various biological effects indifferent types of cells. In addition, we demonstrated
that Smads interact with transcriptional co-activators,
p300 and CBP, and transcriptional co-repressors, c-Ski
and SnoN [26, 31]. c-Ski was originally identified as
an oncogene. Our findings suggest that c-Ski may regulate growth and differentiation of various cells
through regulating the actions of Smads.
We are also studying vascular diseases induced byperturbation of the TGF-β/BMP signaling systems.
Hereditary hemorrhagic telangiectasias (HHTs) are
induced by mutations of the human endoglin or ALK1 genes. ALK-1-null mice exhibit vascular abnormalities reminiscent of human HHT [34]. Primary pulmonary hypertension (PPH) is induced by mutationsof BMPR-II; however, we have shown that mutations

occur in various portions of the BMPR-II in PPH
patients, which may contribute in different fashions
to the pathogenesis of this disease [47]. Using adenovirus vectors containing constitutively active formsof TGF-β type I receptor (ALK-5) or ALK-1, we haveidentified target genes induced by these receptors.
These findings will become useful for our future studies designed to elucidate the mechanisms of pathogenesis of vascular diseases induced by abnormalities
of TGF-β and BMP signaling.

Current Research
Our current research activities are focused on two
major projects. First, we are trying to elucidate thesignaling mechanisms of the TGF-β superfamily
through Smad proteins, and to modulate TGF-β signaling by various molecules. Many tumor cells areresistant to the growth inhibitory activity of TGF-β.
Thus, perturbations of TGF-β superfamily signaling
result in progression of various cancers, including
hereditary non-polyposis colorectal cancer (HNPCC)
and pancreatic cancer (Figure 1). However, the molecular mechanisms of the resistance to effects of TGF-β
are still not fully understood. We have recently
shown that cis-compound disruption of murine
Smad2 gene accelerates malignant progression ofintestinal tumors in Apc knockout mice [49].
Moreover, we have shown that TGF-β regulates transcription of c-myc through Smads [45], and we are
currently studying the molecules that regulate c-myc
transcription together with Smads, including the TCF4/Lef-1 family of transcription factors.
We have shown that I-Smads and c-Ski/SnoN negatively regulate TGF-β signaling. Based on their threedimensional structures, we are trying to elucidate
how these inhibitory molecules interact with the
TGF-β receptors or Smads. We are also trying to identify novel molecules that regulate TGF-β/BMP signaling by yeast two-hybrid systems and DNA microarray
analyses. Experiments using Xenopus embryos which

Figure 2. Differentiation of ES cells into ECs and SMCs in vitro.

61we have recently started in our laboratory will be useful for examining the biological activities regulated bythese molecules. Although perturbations of TGF-β signaling result in malignant progression of tumors,tumor cells by themselves produce large amounts of
TGF-β, which induce progression of tumors throughinduction of neovascularization, inhibition of immune
function, and accumulation of extracellular matrices.
We have recently found that adenoviruses containing
Smad7 and c-Ski prevented metastasis of breast
tumors in nude mice. Thus, molecules that negatively
regulate TGF-β signaling may be useful for regulationof progression and/or metastasis of certain cancers.
Our second project is to study the growth and differentiation of murine ES cells. Formation of new
blood vessels involves the proliferation and differentiation of endothelial progenitor cells (EPCs) circulatingin peripheral blood. We are studying whether TGF-β
superfamily signals play roles during the differentiation of EPCs into endothelial cells (ECs) and smoothmuscle cells (SMCs) using an in vitro ES cell system(Figure 2). Our preliminary findings suggest that thebalance of two major signaling pathways of the TGF-β
superfamily cytokines, i.e. TGF-β-like signals and
BMP-like signals, plays important roles in the differentiation of EPCs into ECs and SMCs. We also foundthat TGF-β regulates differentiation and formation oftight junction of ECs. Moreover, we have obtainedsome evidence that certain fractions of ECs express
specific markers for lymphatic ECs, suggesting thatlymphangiogenesis may be induced from EPCs.

Future Prospects
Our future goals will include further elucidation of
signaling mechanisms of the TGF-β superfamily
cytokines. Inhibition of TGF-β signaling by I-Smads
or transcriptional co-repressors appears to be a powerful method of preventing tissue fibrosis and cancer
metastasis. Development of molecules that regulate
TGF-β signaling pathways may thus be useful fortreatment of various clinical disorders. We will also
extend our research to generate cells of different lineages from murine ES cells. Our knowledge of TGF-β
and other cytokine signaling pathways will be usefulin these projects. Generation of ECs and SMCs fromES cells will be valuable for treatment of various vascular diseases. Moreover, generation of lymphatic ECs
will be useful for identification of genes specifically
expressed in lymphatic tissues. These studies willthus be beneficial not only for application of thesecells in the field of regenerative medicine, but also fordevelopment of new strategies for diagnosis and treatment of metastatic tumors.

62

Research Grants
1. Research for the Future Program: Study on the Signal
Transduction by Serine-Threonine Kinase Receptors(1996-2000) Kohei Miyazono (284,485,000 JPY)
Supported by the Japan Society for the Promotion of
Science.
2. Grant-in-Aid for Scientific Research on Priority Areas(Advanced Research on Cancer): Signaling
Mechanisms of TGF-β and Their Abnormalities in
Human Cancer (2000-2004) Kohei Miyazono(224,270,000 JPY in 2000-2002) Supported by the
Ministry of Education, Culture, Sports, Science, andTechnology of Japan.
3. Grant-in-Aid for Scientific Research (A): Study on the
Biological Activities of Serine-Threonine Kinase
Receptors (2002-2005) Kohei Miyazono (40,900,000
JPY) Supported by the Japan Society for the
Promotion of Science.
4 Grant-in-Aid for Scientific Research (B)(2): Signal
Transduction by Smad Proteins and Their Biological
Activities (1999-2001) Kohei Miyazono (14,900,000
JPY) Supported by the Ministry of Education,
Culture, Sports, Science, and Technology of Japanand the Japan Society for the Promotion of Science.
5. Grant-in-Aid for Scientific Research (B) (2): Signal
Transduction and Biological Activities of theMembers of the TGF-β Superfamily (1997-1998)
Kohei Miyazono (15,100,000 JPY) Supported by the
Ministry of Education, Culture, Sports, Science, andTechnology of Japan.

Select Publications (* indicates 10 selectPublications, for which copies are attached)
1. Miyazono, K., Olofsson, A., Colosetti, P., and Heldin,
C.-H. A role of the latent TGF-b1-binding protein in
the assembly and secretion of TGF-b1. EMBO J. 10,
1091-1101, 1991.
2. ten Dijke, P., Ichijo, H., Franzén, P., Schulz, P., Saras,J., Toyoshima, H., Heldin, C.-H., and Miyazono, K.
Activin receptor-like kinases: A novel subclass of cellsurface receptors with predicted serine/threonine
kinase activity. Oncogene 8, 2879-2887, 1993.
3.* Franzén, P., ten Dijke, P., Ichijo, H., Yamashita, H.,
Schulz, P., Heldin, C.-H., and Miyazono, K. Cloningof a TGFb type I receptor that forms a heteromericcomplex with the TGFb type II receptor. Cell 75,
781-792, 1993.
4. Yamashita, H., Ichijo, H., Grimsby, S., Morén, A., ten
Dijke, P., and Miyazono, K. Endoglin forms a heteromeric complex with the signaling receptors fortransforming growth factor-b. J. Biol. Chem. 269,
1995-2001, 1994.
5. Taipale, J., Miyazono, K., Heldin, C.-H., and KeskiOja, J. Latent transforming growth factor-b1 associates to fibroblast extracellular matrix via latent TGFβ binding protein. J. Cell Biol. 124, 171-181, 1994.
6.* ten Dijke, P., Yamashita, H., Ichijo, H., Franzén, P.,
Laiho, M., Miyazono, K., and Heldin, C.-H.

Characterization of type I receptors for transfoming
growth factor-b and activin. Science 264, 101-104,
1994.
7. ten Dijke, P., Yamashita, H., Sampath, T.K., Reddi,
A.H., Estevez, M., Riddle, D.L., Ichijo, H., Heldin, C.H., and Miyazono, K. Identification of type I receptors for osteogenic protein-1 and bone morphogenetic protein-4. J. Biol. Chem. 269, 16985-16988, 1994.
8. Okadome, T., Yamashita, H., Franzén, P., Morén, A.,
Heldin, C.-H., and Miyazono, K. Distinct roles of theintracellular domains of transforming growth factorb type I and type II receptors in signal transduction.
J. Biol. Chem. 269, 30753-30756, 1994.
9. Yamashita, H., Okadome, T., Franzén, P., ten Dijke,
P., Heldin, C.-H., and Miyazono, K. A rat pituitary
tumor cell line (GH3) expresses type I and type II
receptors and other cell surface binding protein(s) for
transforming growth factor-b. J. Biol. Chem. 270,770-774, 1995.
10.* Yamashita, H., ten Dijke, P., Huylebroeck, D.,
Sampath, T.K., Andries, M., Smith, J.C., Heldin, C.H., and Miyazono, K. Osteogenic protein-1 binds toactivin type II receptors and induces certain activinlike effects. J. Cell Biol. 130, 217-226, 1995.
11. Rosenzweig, B.L., Imamura, T., Okadome, T., Cox,
G.N., Yamashita, H., ten Dijke, P., Heldin, C.-H., andMiyazono, K. Cloning and characterization of ahuman type II receptor for bone morphogenetic proteins. Proc. Natl. Acad. Sci. USA 92, 7632-7636,
1995.
12. Nishitoh, H., Ichijo, H., Kimura, M., Matsumoto, T.,
Makishima, F., Yamaguchi, A., Yamashita, H.,
Enomoto, S., and Miyazono, K. Identification of type
I and type II serine/threonine kinase receptors for
growth/differentiation factor-5. J. Biol. Chem. 271,
21345-21352, 1996.
13. Souchelnytskyi, S., ten Dijke, P., Miyazono, K., andHeldin, C.-H. Phosphorylation of Ser165 in TGF-β
type I receptor modulates TGF-b1-induced cellularresponses. EMBO J. 15, 6231-6240, 1996.
14.* Ichijo, H., Nishida, E., Irie, K., ten Dijke, P., Saitoh,M., Moriguchi, T., Takagi, M., Matsumoto, K.,
Miyazono, K., and Gotoh, Y. Induction of apoptosis
by ASK1, a mammalian MAPKKK that activates
SAPK/JNK and p38 signaling pathways. Science 275,
90-94, 1997.
15. Nakajima, Y., Miyazono, K., Kato, M., Takase, M.,
Yamagishi, T., and Nakamura, H.Extracellular fibrillar structure of latent TGFb binding protein-1: Rolein TGFb-dependent endothelial-mesenchymal transformation during endocardial cushion tissue formation in mouse embryonic heart. J. Cell Biol. 136, 193204, 1997.
16. Nakao, A., Imamura, T., Souchelnytskyi, S.,
Kawabata, M., Ishisaki, A., Oeda, E., Tamaki, K.,
Hanai, J.-i., Heldin, C.-H., Miyazono, K., and ten
Dijke, P. TGF-β receptor-mediated signalling throughSmad2, Smad3 and Smad4. EMBO J. 16, 5353-5362,
1997.

17.* Imamura, T., Takase, M., Nishihara, A., Oeda, E.,
Hanai, J.-i., Kawabata, M. and Miyazono, K. Smad6
inhibits signalling by the TGF-β superfamily. Nature
389, 622-626, 1997.
18.* Heldin, C.-H., Miyazono, K., and ten Dijke, P. TGFβ signalling from cell membrane to nucleus throughSmad proteins. Nature 390, 465-471, 1997.
19. Saitoh, M., Nishitoh, H., Fujii, M., Takeda, K.,
Tobiume, K., Sawada, Y., Kawabata, M., Miyazono,
K., and Ichijo, H. Mammalian thioreodxin is a direct
inhibitor of apoptosis signal-regulating kinase (ASK)
1. EMBO J. 17, 2596-2606, 1998.
20. Takeda, K., Ichijo, H., Fujii, M., Mochida, Y., Saitoh,
M., Nishitoh, H., Sampath, T.K., and Miyazono, K.
Identification of a novel bone morphogenetic protein-responsive gene that may function as a noncoding RNA. J. Biol. Chem. 273, 17079-17085, 1998.
21. Inoue, H., Imamura, T., Ishidou, Y., Takase, M.,
Udagawa, Y., Tsuneizumi, K., Tabata, T., Miyazono,
K., and Kawabata, M. Interplay of signal mediatorsof Decapentaplegic (Dpp): Molecular characterizationof Mothers aganst dpp, Medea, and Daughtersagainst Dpp. Mol. Biol. Cell 9, 2145-2156, 1998.
22.* Kawabata, M., Inoue, H., Hanyu, A., Imamura T.,
and Miyazono, K. Smad proteins exist as monomers
in vivo and undergo homo- and hetero-oligomerization upon activation by serine/threonine kinase
receptors. EMBO J. 17, 4056-4065, 1998.
23. Nishitoh, H., Saitoh, M., Mochida, Y., Takeda, K.,
Nakano, H., Rothe, M., Miyazono, K., and Ichijo, H.
ASK1 is essential for JNK/SAPK activation by TRAF2.
Mol. Cell 2, 389-395, 1998.
24. Yagi, K., Goto, D, Hamamoto, T., Takenoshita, S.,
Kato, M. and Miyazono, K. Alternatively-spliced variant of Smad2 lacking exon 3: Comparison with wildtype Smad2 and Smad3. J. Biol. Chem. 274, 703-709,
1999.
25. Yanagisawa, J., Yanagi, Y., Masuhiro, Y., Suzawa, M.,
Toriyabe, T., Kashiwagi, K., Watanabe, M.,
Kawabata, M., Miyazono, K., and Kato, S.
Convergence of transforming growth factor-b andvitamin D signaling pathways on SMAD transcriptional coactivators. Science 283, 1317-1321, 1999.
26. Nakashima, K., Yanagisawa, M., Arakawa, H.,
Kimura, N., Hisatsune, T., Kawabata, M., Miyazono,
K., and Taga, T. Synergistic signaling in fetal brain
by STAT3-Smad1 complex bridged by p300. Science
284, 479-482, 1999.
27. Nakao, A., Fujii, M., Matsumura, R., Kumano, K.,
Saito, Y., Miyazono, K., and Iwamoto, I. Transient
gene transfer and expression of Smad7 prevents
bleomycin-induced lung fibrosis in mice. J. Clin.
Invest. 104, 5-11, 1999.
28. Nishihara, A., Hanai, J.-i., Imamura, T., Miyazono,
K., and Kawabata, M. E1A inhibits transforming
growth factor-b signaling through binding to Smadproteins. J. Biol. Chem. 274, 28716-28723, 1999.
29.* Fujii, M., Takeda, K., Imamura, T., Aoki, H.,
Sampath, T.K., Enomoto, S., Kawabata, M., Kato, M.,
Ichijo, H., Miyazono, K. Roles of bone morphogenetic

63

protein type I receptors and Smad proteins in
osteoblastic and chondroblastic differentiation. Mol.
Biol. Cell 10, 3801-3813, 1999.
30. Hanai, J.-i., Chen, L.F., Kanno, T., Ohtani-Fujita, N.,
Kim, W.Y., Guo, W.-H., Imamura, T., Ishidou, Y.,
Fukuchi, M., Shi, M.-J., Stavnezer, J., Kawabata, M.,
Miyazono, K., and Ito, Y. Interaction and functional
cooperation of PEBP2/CBF with Smads: Synergistic
induction of the immunoglobulin germline Ca promoter. J. Biol. Chem. 274, 31577-31582, 1999.
31. Akiyoshi, S., Inoue, H., Hanai, J.-i., Kusanagi, K.,
Nemoto, N., Miyazono, K., and Kawabata, M. c-Ski
acts as a transcriptional co-repressor in TGF-β signaling through interaction with Smads. J. Biol. Chem.
274, 35269-35278, 1999.
32. Kusanagi, K., Inoue, H., Ishidou, Y., Mishima, H.K.,
Kawabata, M., and Miyazono, K. Characterization ofa bone morphogenetic protein-responsive Smadbinding element. Mol. Biol. Cell 11, 555-565, 2000.
33. Ishida, W., Hamamoto, K., Kusanagi, K., Yagi, K.,
Kawabata, M., Takehara, K., Sampath, T.K., Kato,
M., and Miyazono, K. Smad6 is a Smad1/5-induced
Smad inhibitor: Characterization of bone morphogenetic protein-responsive element in the mouse
Smad6 promoter. J. Biol. Chem. 275, 6075-6079,
2000.
34. Oh, S.P., Seki, T., Goss, K.A., Imamura, T., Yi, Y.,
Donahoe, P.K., Li, L., Miyazono, K., ten Dijke, P.,
Kim, S., and Li, E.) Activin receptor-like kinase 1
modulates transforming growth factor-b1 signaling
in the regulation of angiogenesis Proc. Natl. Acad.
Sci. USA 97, 2626-2631, 200.
35. Miyazono, K. Positive and negative regulation of
TGF-β signaling. J. Cell Sci. 113, 1101-1109, 2000.
36. Beppu,, H., Kawabata, M., Hamamoto, T., Chytil, A.,
Minowa, O., Noda, T., and Miyazono, K. BMP type II
receptor is required for gastrulation and early development of mouse embryos. Dev. Biol. 221, 249-258,
2000.
37. Zhang, Y.-W., Yasui, N., Huang, G., Fujii, M., Hanai,
J.-i., Nogami, H., Ochi, T., Miyazono, K., and Ito, Y.
A RUNX2/PEBP2αA/CBFA1 mutation displaying
impaired transactivation and Smad interaction incleidocranial dysplasia. Proc. Natl. Acad. Sci. USA 97,
10549-10554, 2000.
38. Miura, S., Takeshita, T., Asao, H., Kimura, Y.,
Murata, K., Sasaki, Y., Hanai, J.-i., Beppu, H.,
Tsukazaki, T., Wrana, J.L., Miyazono, K., andSugamura, K. Hgs (Hrs), a FYVE domain protein, isinvolved in Smad signaling through cooperation with
SARA. Mol. Cell. Biol. 20, 9346-9355, 2000.
39. Yoshida, Y., Tanaka, S., Umemori, H., Minowa, M.,
Usui, M., Ikematsu, N., Hosoda, E., Imamura, T.,
Kuno, J., Yamashita, T., Miyazono, K., Noda, M.,
Noda, T., and Yamamoto, T. Negative regulation of
BMP/Smad signaling by Tob in osteoblasts. Cell 103,1085-1097, 2000.
40.* Ebisawa, T., Fukuchi, M., Murakami, G., Chiba, T.,
Tanaka, K., Imamura, T., and Miyazono, K. Smurf1
interacts with transforming growth factor-b type I

64

receptor through Smad7 and induces receptor degradation. J. Biol. Chem. 276, 12477-12480, 2001.
41. Fukuchi, M., Imamura, T., Chiba, T., Ebisawa, T.,
Kawabata, M., Tanaka, K., and Miyazono, K.Liganddependent degradation of Smad3 by a ubiquitin ligase complex of ROC1 and associated proteins. Mol.
Biol. Cell 12, 1431-1443, 2001.
42. Furuhashi, M.,Yagi, K.,Yamamoto, H., Furukawa, Y.,
Shimada, S., Nakamura, Y., Kikuchi, A., Miyazono,
K., and Kato, M. Axin facilitates Smad3 activation in
the transforming growth factor-b signaling pathway.
Mol. Cell. Biol. 21, 5132-5141, 2001.
43. Kusanagi, K., Kawabata, M., Mishima, H.K., andMiyazono, K. a-Helix 2 in the amino-terminal Mad
homology 1 domain is responsible for specific DNAbinding of Smad3. J. Biol. Chem. 276, 28155-28163,
2001.
44.* Hanyu, A., Ishidou, Y., Ebisawa, T., Shimanuki, T.,
Imamura, T., and Miyazono, K. The N-domain of
Smad7 is essential for specific inhibition of transforming growth factor-b signaling. J. Cell Biol. 155,
1017-1028, 2001.
45. Yagi, K., Furuhashi, M., Aoki, H., Goto, D., Kuwano,
H., Sugamura, K., Miyazono, K., and Kato, M. c-myc
is a downstream target of Smad pathway. J. Biol.
Chem. 277, 854-861, 2002.
46. Ring, C., Ogata, S., Meek, L., Song, J., Ohta, T.,
Miyazono, K., and Cho, K.W.Y. The role of a
Williams-Beuren syndrome-associated helix-loophelix domain-containing transcription factor in
activin/nodal signaling. Genes Dev. 16, 820-835,
2002.
47. Nishihara, N., Watabe, T., Imamura, T., andMiyazono, K. Functional heterogeneity of BMPR-II
mutants found in patients with primary pulmonary
hypertension. Mol. Biol. Cell 22, 5212-5221, 2002.
48. Suzuki, C., Murakami, G., Fukuchi, M., Shimanuki,T., Shikauchi, Y., Imamura, T., and Miyazono, K.
(2002) Smurf1 regulates the inhibitory activity of
Smad7 by targeting Smad7 to the plasma membrane.
J. Biol. Chem. in press.
49. Hamamoto, T., Beppu, H., Okada, H., Kawabata, M.,
Kitamura, T., Miyazono, K., and Kato, M. (2002)
Compound disruption of Smad2 accelerates malignant progression of intestinal tumors in Apc knockout mice. Cancer Res. in press.
50. Miyazono, K., and Miyazawa, K. (2002) Id: A target
of BMP signaling. Science’s STKE in press.

Department ofMicrobiology
Outline and Research Objectives
Microbial disease has been recognized as the major threat to human health throughout
the history. Despite the development of preventive and therapeutic interventions against
some pathogenic microbes, infectious disease is still one of the most significant medical
problems. On the other hand, microbial organisms have served as a useful model as well for
elucidating the human molecular mechanisms of a variety of biological events, providing
useful insights into life science. Efforts have also been initiated by a number of researchgroup to utilize animal viruses as a tool for human gene therapy. In order to familialize students with these issues, importance of microbiology in medical education is increasing more
rapidly than ever. To fulfill this requirement, our department, as the only basic microbiology
unit in the Faculty and Graduate School, currently assumes a responsibility for teaching bacteriology and virology to medical undergraduates, and for educating graduate students in afield of virology on animal RNA viruses.
Our major research interest is to elucidate molecular mechanisms for species-, tissue-,and cell-specific pathogenicity of poliovirus and hepatitis C virus. For this purpose, we
attempt to identify viral and host factors required for the viral replication and disseminationin animal model. Tissue and cell distribution of host factors in host species or animal model
may give an insight into molecular pathogenesis of those RNA viruses. Thus, the main
objectives in our department are as follows;
I. Replication, dissemination, and neurovirulence of poliovirus
Poliovirus (PV), the causative agent of poliomyelitis, is a human enterovirus that belongs
to the Picornaviridae. The genome of PV is a single-stranded RNA of positive polarity andconsists of approximately 7500 nucleotides (nt). To this RNA, a small protein VPg isattached at the 5’end, and poly(A) at the 3’ end. PV mRNA does not have VPg and starts
with simple pU. Thus PV mRNA is an exceptional eukaryotic mRNA. An internal ribosome
entry site (IRES) was identified in the 5’ untranslated region (UTR) of the RNA.
Natural PV infection in humans is thought to begin with oral ingestion. After ingestion,
the virus multiplies in the alimentary mucosa; the tonsils and Peyer’s patches are possibly
also invaded early in the course of infection. Following multiplication in these loci, the virus
moves into the deep cervical and mesenteric lymph nodes and then into the blood.
Neutralizing antibodies to PV in the blood prevent the development of poliomyelitis, whichsuggests that viremia is necessary for the spread of virus to the central nervous system(CNS). The circulating virus then invades the CNS and replicates in neurons, particularly the
motor neurons. There are two possible routes through which PV can enter the CNS. One isvirus permeation through the blood-brain barrier (BBB) and the other is virus transmission
via peripheral nerves. Paralytic poliomyelitis occurs as a result of neuronal destruction.
Specifically, we are working on the following projects.
1. Roles of human PV receptor (hPVR) in PV infection.
2. Molecular mechanisms for BBB permeation of PV.
3. Molecular mechanisms for retrograde axonal transport of PV.
4. Replication of PV in neurons.
II. Replication and pathogenesis of hepatitis C virus
Hepatitis C virus (HCV), the main causative agent of non-A, non-B hepatitis, is a memberof Flaviviridae. Chronic infection of HCV frequently leads to liver cirrhosis and hepatocellular carcinoma (HCC). The genome of HCV is a single-stranded RNA of positive polarity andconsists of approximately 9500 nt. An IRES was found in the 5’ portion of the HCV RNA
like PV RNA, although it is not known whether the 5’ end is capped or not. As for the 3’65UTR, the HCV RNA does not have a poly(A) tail but a poly(U/C) tract of 80-130 nt followed
by a highly conserved 98 nt sequence among HCV isolates, termed the X region. Thus, HCV
RNA has an unique structure as an eukaryotic mRNA. Specifically, we are working on thefollowing projects.
1. Molecular mechanisms for development of HCC by HCV infection.
2. Replication mechanisms of HCV RNA replicon.
III. IRES-dependent virus tropism
Virulent strains of PV can replicate well both in the CNS and the alimentary mucosa,
whereas attenuated strains of PV cannot replicate well in the CNS although these strains
have a strong replicating capacity in the alimentary mucosa. Therefore, virulent and attenuated PV strains have different tissue tropism each other. Relatively strong determinant for
the attenuation phenotype was identified in the 5’ UTR within the IRES. It is possible that
IRES activity differ in tissues, and that viral replication capacity reflects the IRES activity.
IRES was first described for picornavirus RNAs, and later for many cellular mRNAs as
well as other viral RNAs. Nucleotide sequences that serve as IRES, so far discovered, have avariety of lengths and predicted secondary structures, although all of them have a similarfunction in translation initiation. Existence of a variety of IRES structures suggests that different sets of trans-acting cellular factors are required for activity of individual IRESs.
Cumulative evidence suggests that cellular factors required for IRES activities are quantitatively and/or qualitatively different in individual IRESs. Specifically, we are working on thefollowing projects.
1. Construction of recombinant viruses that have mutated IRESs.
2. Identification of host factors required for activity of various IRESs.
3. Establishment of method for measuring IRES activity in vivo.

Faculties and Students
Professor and Chair Akio Nomoto, Ph.D. (since 1999)
Associate Professor Tetsuro Matano, M.D., Ph.D.
Associate .................................2
Postdoctoral Fellow ................3
Graduate Students ................12
Secretary .................................2

Past Research and Major Accomplishments
1. Structural analysis of the 5’ termini of all the PV
specific RNAs and proposal of VPg-primer theoryof PV RNA synthesis.
2. Determination of the primary structure of the
genomes of all three attenuated PV serotypes
3. Construction of infectious cDNA clone of theattenuated PV strain (Sabin 1 strain) and first genemanipulation of animal RNA virus using the infectious clone.
4. Proposal of new copy choice model of RNA
genome for its deletion and recombination events.
5. Molecular genetic analysis of the attenuation phenotype of type 1 PV.
6. Construction of new attenuated PV strains basedon knowledge of stability of attenuation mutations.
7. Discovery of hPVR.

66

8. Establishment of hPVR/transgenic (Tg) mice susceptible to PV.
9. Analysis of interaction between PV and hPVR.
10. Formation of a concept “IRES-dependent virus tropism”.
11. Study on dissemination pathway of PV using Tgmouse model.
12. Subcellular localization of hPVR in polarized cells.
13. Response of neural cells to poliovirus infection.
14. Discovery of HCV IRES

Current Research
1. There are two isoforms of functional human PV
receptors, hPVRα and hPVRδ. These two isoforms differ only in a part of amino acid sequence
within the cytoplasmic domain. Basolateral sorting signal was identified in the cytoplasmic
domain of hPVRα but not hPVRδ. Indeed, hPVR
αlocalizes to the basolateral membran and hPVR
δ apical membrane. It is possible that PV infection occurs by using hPVRδin the human alimentary tract.
2. Using a hPVR/Tg mouse model, we have shownthat PV permeates the BBB at a fairly high rate,
independently of the presence of hPVR or virus
strain-specific effects. Thus, some host cell molecules other than hPVR must be involved in the
BBB permeability of PV. We are currently investi-

3.

4.

5.

6.

gating which host molecules are involved in thisdissemination process.
Our recent results strongly suggest the following;
Intramuscularly inoculated PV is incorporated by
hPVR-mediated endocytosis at synapses without
conformational change. The endosomes containing PV is associated with dynein motor molecule,and retrogradely transported to neuron cell bodyalong microtubules. After the retrograde transport
through the axon, lytic replication of PV occurs in
the cell body. The virulent and attenuated PV
strains do not show significant difference in efficiency of this dissemination route in a hPVR/Tg
mouse model. We are attempting to prove theabove bypothesis.
PV-infected neural cells do not show cytopathic
effect when the infected cell cultures are addedwith anti-PV or anti-PVR antibodies 2 hours after
the infection. Similar phenomenon cannot be
observed in PV-infected HeLa cells. We precisely
analyzed this phenomenon, and found that PV
IRES activity was inhibited in the infected neuralcells, and that PV-specific 2A protease was transported to the nucleus, and therefore translation
initiation factor eIF4G was restored in the infected
cells. We are currently attempting to elucidate
why PV IRES activity is inhibited in the infectedneural cells.
Development of cancers always accompanies
mutations in chromosomes. Although HCV
genome does not have any specific tumor gene, it
is possible that some HCV gene products induce
instability of human chromosome structure. We
found that nuclear transport system was disturbed
by HCV core protein. This may result in instabilityof human chromosome structure in the HCVinfected hepatocytes. We are now investigatinginteraction between HCV core protein and nuclear
transport receptors.
Chimeric PV whose IRES was replaced by HCVIRES was constructed. This recombinant virus canreplicate well in the liver of hPVR/Tg mice but notin the brain of the mice. This indicates that HCVIRES is active in the liver but not in the brain. The
results support our notion “ IRES-dependent virus
tropism”.

Future Prospects
PV is now one of the most well characterized virus
and recent studies of its molecular biology have madegreat progress. Additionally, the development of atransgenic mouse model for poliomyelitis has not
only facilitated investigations of PV dissemination but
has transformed PV research into an important area ofneuroscience. The elucidation of the molecular basisof PV neurovirulence is only at its starting point, andsome basic questions proposed in section of “ Current
Research” must be answered if we are to understandfully the molecular mechanisms responsible. This
research could eventually lead to the development of
new strategies to control poliomyelitis and other viraldiseases.
It is an important question in life science whyviruses without tumor gene are able to cause cancers.
Extensive effort have been made to elucidate mechanisms for development of HCC by HCV infection.
However, little is known, as yet, about the mechanisms responsible. Since development of effective vaccine to control HCV infection seems to be difficult,and altogether may be impossible, only sure way tocontrol HCV is to elucidate molecular mechanisms
responsible for onset of HCV disease, resulting in thedevelopment of new strategies to control the viral disease.
Our approach to study pathogenicity of RNA viruses may also provide useful insights into general life
sciences.

Research Grants
1995-1999 Grant-in-aid for Specially Promoted Research(total ¥273,000,000)
2000-2004 Grant-in-aid for Specially Promoted Research(~2002 ¥182,000,000)
2001-2005 Research Grant from the Organization for
Pharmaceutical Safety and Research (~2002¥65,000,000)

Select Publications
1. Akio Nomoto, Yuan Fon Lee & Eckard Wimmer. The
5’ end of poliovirus mRNA is not capped with
m7G(5’)ppp(5’)Np.
Proc. Natl. Acad. Sci. USA 73: 375-380, 1976
2. Yuan Fon Lee, Akio Nomoto, Barbara M. Detjen &Eckard Wimmer. A protein covalently linked topoliovirus genome RNA.
Proc. Natl. Acad. Sci. USA 74(1): 59-63, 1977
3. Akio Nomoto, Barbara M. Detjen, Rudy Pozzatti &Eckard Wimmer. The location of the polio genome
protein in viral RNAs and its implication for RNA
synthesis.
Nature 268(5617): 208-213, 1977
4. Akio Nomoto, Naomi Kitamura, Fred Golini &Eckard Wimmer. The 5’-terminal structures ofpoliovirus RNA and poliovirus mRNA differ only in
the genome-linked protein VPg.
Proc. Natl. Acad. Sci. USA 72(12): 5345-5349, 1977
5. Akio Nomoto, Ann Jacobson, Yuan Fon Lee, John
Dunn & Eckard Wimmer. Defective interfering particles of poliovirus: Mapping of the deletion and evi-67

dence that the deletions in the genomes of DI (1), (2)and (3) are located in the same region.
J. Mol. Biol. 128(2): 179-196, 1979
6. Akio Nomoto, Toshiko Omata, Haruka Toyoda,
Shusuke Kuge, Hitoshi Horie, Yutaka Kataoka, Yuko
Genba, Yasuko Nakano & Nobumasa Imura.
Complete nucleotide sequence of the attenuatedpoliovirus Sabin 1 strain Genome.
Proc. Natl. Acad. Sci. USA 79(19): 5793-5797, 1982
7. Haruka Toyoda, Michinori Kohara, Yutaka Kataoka,
Toshiyuki Suganuma, Toshiko Omata, Nobumasa
Imura & Akio Nomoto. Complete nucleotide
sequences of all three poliovirus serotype genomes:
Implication for genetic relationship, gene function
and antigenic determinants.
J. Mol. Biol. 174(4): 561-585, 1984
8. Toshiko Omata, Michinori Kohara, Yuko Sakai,
Atsuko Kameda, Nobumasa Imura & Akio Nomoto.
Cloned infectious complementary DNA of poliovirus
Sabin 1 genome: biochemical and biological properties of the recovered virus.
Gene 32 (Part I): 1-10, 1984
9. Michinori Kohara, Toshiko Omata, Atsuko Kameda,
Bert L. Semler, Heihachi Itoh, Eckard Wimmer &Akio Nomoto. In vitro phenotypic markers of apoliovirus recombinant constructed from infectious
cDNA clones of the neurovirulent Mahoney andattenuated Sabin 1 strains.
J. Virol. 53(3): 786-792, 1985
10. Toshiko Omata, Michinori Kohara, Shusuke Kuge,
Toshihiko Komatsu, Shinobu Abe, Bert L. Semler,
Atsuko Kameda, Heihachi Itoh, Mineo Arita, Eckard
Wimmer & Akio Nomoto. Genetic analysis of theattenuation phenotype of poliovirus type 1.
J. Virol. 58(2): 348-358, 1986
11. Michinori Kohara, Shinobu Abe, Shusuke Kuge, Bert
L. Semler, Toshihiko Komatsu, Mineo Arita,
Heihachi Itoh & Akio Nomoto. An infectious cDNA
clone of the poliovirus Sabin strain could be used asa stable repository and inoculum for the oral polio
live vaccine.
Virology 151(1): 21-30, 1986
12. Shusuke Kuge, Izumu Saito & Akio Nomoto. Primarystructure of poliovirus defective interfering particle
genomes and possible generation mechanisms of theparticle.
J. Mol. Biol. 192: 473-487, 1986
13. Shusuke Kuge & Akio Nomoto. Construction of
viable deletion and insertion mutants of the Sabin
strain of type 1 poliovirus: Function of the 5’ noncoding sequence in viral replication.
J. Virol. 61(5): 1478-1487, 1987
14. Akio Nomoto, Narushi Iizuka, Michinori Kohara &Mineo Arita. Strategy for construction of live picornavirus vaccines.
Vaccine 6(2): 134-137, 1988
15. Michinori Kohara, Shinobu Abe, Toshihiko Komatsu,
Katsuhiko Tago, Mineo Arita & Akio Nomoto. A
recombinant virus between the Sabin 1 and Sabin 3

68

vaccine strains of poliovirus as a possible candidate
for a new type 3 poliovirus live vaccine strain.
J. Virol. 62(8): 2828-2835, 1988
16. Shusuke Kuge, Noriyuki Kawamura & Akio Nomoto.
Genetic variation occurring on the genome of an invitro insertion mutant of poliovirus type 1.
J. Virol. 63(3): 1069-1075, 1989
17. Noriyuki Kawamura, Michinori Kohara, Shinobu
Abe, Toshihiko Komatsu, Katsuhiko Tago, Mineo
Arita & Akio Nomoto. Determinants in the 5’ noncoding region of poliovirus Sabin 1 RNA that influence the attenuation phenotype.
J. Virol. 63(3): 1302-1309, 1989
18. Shusuke Kuge, Noriyuki Kawamura & Akio Nomoto.
Strong inclination toward transition mutation in
nucleotide substitutions by poliovirus replicase.
J. Mol. Biol. 207(1): 175-182, 1989
19. Narushi Iizuka, Michinori Kohara, Kimiko HaginoYamagishi, Shinobu Abe, Toshihiko Komatsu,
Katsuhiko Tago, Mineo Arita & Akio Nomoto.
Construction of less neurovirulent polioviruses byintroducing deletions into the 5’ noncoding sequenceof the genome.
J. Virol. 63(12): 5354-5363, 1989
20. Kimiko Hagino-Yamagishi & Akio Nomoto. In vitro
construction of poliovirus defective-interfering particles.
J. Virol. 63(12): 5386-5392, 1989
21. Satoshi Koike, Hitoshi Horie, Iku Ise, Akira Okitsu,
Michihiro Yoshida, Narushi Iizuka, Kenji Takeuchi
Tsutomu Takegami & Akio Nomoto. The poliovirus
receptor protein is produced both as membranebound and secreted forms.
EMBO J. 9(10): 3217-3224, 1990
22. Satoshi Koike, Choji Taya, Takeshi Kurata, Shinobu
Abe, Iku Ise, Hiromichi Yonekawa & Akio Nomoto.
Transgenic mice susceptible to poliovirus.
Proc. Natl. Acad. Sci. USA 88(3): 951-955, 1991
23. Kyoko Tsukiyama-Kohara, Michinori Kohara, Kenjiro
Yamaguchi, Noboru Maki, Ayumi Toyoshima,
Keizaburo Miki, Satoshi Tanaka, Nobu Hattori &Akio Nomoto. A second group of hepatitis C virus.
Virus Genes 5(3): 243-254, 1991
24. Satoshi Koike, Iku Ise & Akio Nomoto. Functional
domains of the poliovirus receptor.
Proc. Natl. Acad. Sci. USA 88(5): 4104-4108, 1991
25. Narushi Iizuka, Hiromichi Yonekawa & Akio
Nomoto. Nucleotide sequences important for translation initiation of enterovirus RNA.
J. Virol. 65(9): 4867-4873, 1991
26. Kyoko Tsukiyama-Kohara, Narushi Iizuka, Michinori
Kohara & Akio Nomoto. Internal ribosome entry site
within hepatitis C virus RNA.
J. Virol. 66(3): 1476-1483, 1992
27. Satoshi Koike, Iku Ise, Yasuko Sato, Kotoko Mitsui,
Hitoshi Horie, Hideaki Umeyama & Akio Nomoto.
Early events of poliovirus infection.
Seminars in Virology, Vol.3, 1992, Cellular Receptors
for Virus Infection (A. Nomoto, ed.) pp109-115,
Saunders Scientific Publications/ Academic Press28. Satoshi Koike, Iku Ise, Yasuko Sato, Hiromichi
Yonekawa, Osamu Gotoh & Akio Nomoto. A second
gene for African green monkey poliovirus receptor
that has no putative N-glycosylation site in the functional N-terminal immunoglobulin-like domain.
J. Virol. 66(12): 7059-7066, 1992
29. Akio Nomoto. Recombinant polioviruses as candidates for oral poliovaccines.
Microbiol. Immunol. 37(3): 169-174, 1993
30. Akio Nomoto, Satoshi Koike & Junken Aoki. Tissue
tropism and species specificity of poliovirus infection.
Trends in Microbiology, Vol. 2, No.2, 47-51, 1994
31. Hitoshi Horie, Satoshi Koike, Takeshi Kurata,
Yasuko Sato-Yoshida, Iku Ise, Yoshihiro Ota,
Shinobu Abe, Kyoji Hioki, Hideki Kato, Choji Taya,
Tatsuji Nomura, Sou Hashizume, Hiromichi
Yonekawa & Akio Nomoto. Transgenic mice carrying
the human poliovirus receptor: new animal model
for study of poliovirus neurovirulence.
J. Virol. 68 (2): 681-688, 1994
32. Junken Aoki, Satoshi Koike, Iku Ise, Yasuko SatoYoshida & Akio Nomoto. Amino acid residues onhuman poliovirus receptor involved in interaction
with poliovirus.
J. Biol. Chem. 269 (11) : 8431-8438, 1994
33. Shinobu Abe, Yoshihiro Ota, Satoshi Koike, Takeshi
Kurata, Hitoshi Horie, Tatsuji Nomura, So
Hashizume & Akio Nomoto. Neurovirulence test fororal live poliovaccines using poliovirus-sensitivetransgenic mice.
Virology 206: 1075-1083, 1995
34. Kazuko Shiroki, Toshihiko Ishii, Takahiro Aoki,
Yoshihiro Ohta, Wei-Xing Yang, Toshihiko Komatsu,
Yasushi Ami, Mineo Arita, Shinobu Abe, So
Hashizume & Akio Nomoto. Host range phenotype
induced by mutations in the internal ribosomal entry
site of poliovirus RNA.
J. Virol. 71 (1): 1-8, 1997
35. Wei-Xing Yang, Tetsuya Terasaki, Kazuko Shiroki,Seii Ohka, Junken Aoki, Shuichi Tanabe, Tatsuji
Nomura, Eiji Terada, Yuichi Sugiyama & Akio
Nomoto. Efficient delivery of circulating poliovirus tothe central nervous system independently ofpoliovirus receptor.
Virology 229 (2): 421-428, 1997
36. Nobuhiko Kamoshita, Kyoko Tsukiyama-Kohara,
Michinori Kohara & Akio Nomoto. Genetic analysis
of internal ribosomal entry site on hepatitis C virus
RNA: implication for involvement of the highlyordered structure and cell type specific trans-acting
factors.
Virology 223 (1): 9-18, 1997
37. Junken Aoki, Satoshi Koike, Hiroaki Asou, Iku Ise,
Hiroshi Suwa, Toshiyuki Tanaka, Masayuki
Miyasaka & Akio Nomoto. Mouse homolog ofpoliovirus receptor-related gene 2 product, mPRR2,
mediates cell aggregation.
Exp. Cell Res. 235: 374-384, 1997

38. Minetaro Arita, Satoshi Koike, Junken Aoki, Hitoshi
Horie & Akio Nomoto. Interaction of poliovirus with
its purified receptor and conformational alteration in
the virion.
J. Virol. 72 (5): 3578-3586, 1998
39. Seii Ohka, Wei-Xing Yang, Eiji Terada, KunikoIwasaki & Akio Nomoto. Retrograde transport ofintact poliovirus through the axon via the fast transport system.
Virology 250 (1): 67-75, 1998
40. Minetaro Arita, Hitoshi Horie, Mineo Arita & Akio
Nomoto. Interaction of poliovirus with its receptor
affords a high level of infectivity to the virion inpoliovirus infections mediated by the Fc receptor.
J. Virol. 73(2): 1066-1074, 1999
41. Kazuko Shiroki, Takeshi Isoyama, Shusuke Kuge,
Toshihiko Ishii, Shinobu Ohmi, Syoji Hata, Koichi
Suzuki, Yoshinari Takasaki & Akio Nomoto.
Intracellular redistribution of truncated La protein
produced by poliovirus 3Cpro-mediated cleavage.
J. Virol.73(3): 2193-2200, 1999
42. Qingmei Jia, Seii Ohka, Kuniko Iwasaki & Akio
Nomoto. Isolation and molecular characterization ofa poliovirus type 1 mutant that replicates in the
spinal cords of mice.
J. Virol. 73(7): 6041-6047, 1999
43. Takeshi Isoyama, Nobuhiko Kamoshita, Kotaro
Yasui, Atsushi Iwai, Kazuko Shiroki, Haruka Toyoda,
Akio Yamada & Akio Nomoto. Lower concentrationof La protein required for internal ribosome entry on
hepatitis C virus than on poliovirus RNA.
J. Gen. Virol. 80(9): 2319-2327, 1999
44. Kyoko Murakami, Michikazu Abe, Tsutomu
Kageyama, Nobuhiko Kamoshita & Akio Nomoto.
Down regulation of translation driven by hepatitis C
virus internal ribosomal entry site by the 3’ untranslated region of RNA.
Arch. Virol. 146(4): 729-741, 2001
45. Qingmei Jia, James M. Hogle, Tsutomu Hashikawa &Akio Nomoto. Molecular genetic analysis of revertants from a poliovirus mutant that is specifically
adapted to the spinal cord of mice.
J. Virol., 75(23): 11766-11772, 2001
46. Seii Ohka, Hitoshi Horie, Koujiro Tohyama & Akio
Nomoto. Basolateral sorting of human poliovirus
receptor a involves an interaction with the m1B subunit of the clathrin adaptor complex in polarized
epithelial cells.
Biochem. Biophys. Res. Comm., 287: 941-948, 2001
Seii Ohka & Akio Nomoto. Recent insights into
poliovirus pathogenesis.
Trends in Microbiology, 9(10): 501-506, 2001
48. Qingmei Jia, Fengyi Liang, Seii Ohka, Akio Nomoto& Tsutomu Hashikawa. Expression of brain-derived
neurotrophic factor in the central nervous system ofmice using a poliovirus-based vector.
J. NeuroVirology, 8(1): 14-23, 2002
49. Akio Nomoto & Isao Arita. Eradication of
Poliomyelitis
Nature Immunology, 3(3): 205-208, 2002

69

50. Takeshi Isoyama, Shusuke Kuge & Akio Nomoto.
The core protein of hepatitis C virus is imported into
the nucleus by transport receptor Kap123p but
inhibits Kap121p-dependent nuclear import of yeastAP1-like factor in yeast cells.
J. Biol. Chem., 277(42): 39634-39641, 2002
Publications indicated by bold letters are attached.

70Department of Infection
Control and Prevention
Outline and Research Objectives
The Department of Infection Control and Prevention was founded in 1994 in order tominimize hospital infections. This department is, therefore, very new, but is the oldest one
in the field of infection control in Japan. It started at first as the Division of Hospital
Infection and Control Services on January 23, 1991. This division developed into theDivision of Infection Control and Prevention on September 1, 1993 and the present department on June 24, 1994.
Research and practice of hospital infection control in Japan has been very primitives, andat least 15-20 years behind those in USA and European countries. The main objectives ofour department are; 1) to control and prevent hospital infections, 2) to develop and propagate new effective methods to control and prevent hospital infections and 3) to be a modeland lead of infection control in Japan. Therefore, surveillance of hospital infections, early
detection of outbreaks, analyses of the causes of outbreaks and intervention are the mostimportant parts of our activities. Education of medical and comedical staffs and students is
also a very important role of our department.

Faculties and Students
Professor and Chair Satoshi Kimura, MD, PhD (1996〜)
Lecturer (Hospital) Kyoji Moriya, MD, PhD (2002〜)
Associate (Hospital) ................2
Postdoctoral Fellow ................1
Research student.....................2
Secretary .................................2

Past Research and Major Accomplishments
1. Establishment of hospital infection surveillancesystem in the University of Tokyo Hospital (comprehensive and targeted).
2. Reduction of surgical site infections by replacing
razor with clipper for preoperational removal ofbody hair at surgical sites.
3. Introduction of EPINet system into Japan and conduction of nationwide surveillance of needlestick
injuries in health care workers in Japan.
4. Reduction of needlestick injuries among healthcare workers by employing safety devices.
5. Epidemiological analyses of Methicillin-resistant
Staphylococcus aureus (MRSA) and other drug
resistant microbes by using PFGE and/or PCR in
the University of Tokyo Hospital.
6. Control of MRSA outbreak in NICU by unselected
use of mupirocin ointment.
7. Control of Ralstonia pickettii colonization ofpatients in an obstetric ward.
8. Evaluation of disinfectant and antiseptics.
9. Nationwide surveillance of opportunistic infections in HIV infection.

10. Development of PCR systems for diagnosis ofopportunistic infections in HIV/AIDS.
11. Quantitative analyses of cytomegalovirus (CMV) inthe blood using real-time PCR for early diagnosis,
monitoring and prospecting of CMV infections.
12. Evaluation of clinical efficacy of antiretroviral
agents and highly active antiretroviral therapy(HAART).
13. Pharmacodynanic analyses of anti-HIV agents.
14. Evaluation of clinical significance of drug-resistance mutations of HIV against protease inhibitors.
15. Analyses of hospital visits among HIV-infected
persons and AIDS cases in Japan.
16. Anti-HIV activities of oligodeoxyribonucleotides,
ribozymes, fluoroquinolones etc..
17. Elucidation of molecular mechanism of viral hepatocarcinogenesis (HBV and HCV).
18. Elucidation of molecular mechanism of chemotaxis.
19. Elucidation of mode of action of defensin andcloning of a nevel mouse β-defensin.
20. Establishment of certification system of Infection
Control Doctors (ICD) in Japan by organizing
Japanese Council of ICD consisting of 16 scientificsocieties in the field of infectious diseases.

Current Research
We have been mainly studying on following subjects:
1. Surveillance, analyses of hospital infections, andplanning to reduce hospital infections and occupational infections, such as needle stick injuries.

712. Epidemiological analyses of methicillin-resistant
Staphylococcus aureus and other drug-resistantmicrobes using PFGE, AP-PCR, etc.
3. Evaluation of disinfectants and antiseptics.
4. Development of sensitive and rapid methods todetect pathogens.
5. Evaluation of multiple-drug therapies for HIV
infection.
6. Quantification of DNA viruses in the blood using
real-time PCR.
7. Preparation of guideline for appropriate use of
antimicrobials.
8. Molecular mechanism o viral hepatocarcinogenesis.

Future Prospect
Surveillance is most important for the analysis ofcauses of outbreaks, and for the development of new
preventive measures. So far, we have established surveillance system in the University of Tokyo Hospital.
ICD system established by Prof. S. Kimura in 1999
greatly stimulated the trend to start surveillance inmany other hospitals all over Japan. Thus, level of
infection control and prevention in Japan will be rapidly elevated. In this process, staffs in our department
will gain initiative.
By reducing hospital infections, duration of hospital stay of patients is to be shortened, medical
expense be reduced, prognosis of patients be better,and quality of life be improved.

Research Grants
1. 1998：Grant-in-Aid from the Ministry of Health andWelfare of Japan（H-9-AIDS-002） Studies on HIV
Infection” ¥216,400,000
2. Grant-in-Aid from the Ministry of Health andWelfare of Japan Studies on Epidemiology of HIV”
¥500,000
3. Grant-in-Aid
forInternational
MedicalCooperation“Studies on International Medical
Cooperation against HIV Pandemic” ¥22,000,000
4. Grant-in-Aid from the Organization for
Pharmaceutical Safety and Research“Epidemiologicaland Clinical Studies on Complications of HIV
Infections” ¥90,000,000
5. 1999：Grant-in-Aid from the Ministry of Health andWelfare of Japan（H-9-AIDS-002） Studies on HIV
Infection” ¥216,426,000
6. Grant-in-Aid from the Ministry of Health andWelfare of Japan Studies on Epidemiology of HIV”
¥500,000
7. Grant-in-Aid from the Organization for
Pharmaceutical
SafetyandResearch
Epidemiological and Clinical Studies on
Complications of HIV Infections” ¥90,000,00072

8. 2000：Grant-in-Aid from the Ministry of Health,
Labor and Welfare of Japan（H-12-AIDS-004）
Studies on the Management of Complication in
HIV/AIDS” ¥70,000,000
9. Grant-in-Aid from the Ministry of Health, Labor andWelfare of Japan Socio-epidemiological Studies on
the Trends of HIV/AIDS in Japan and Its
Intervention” ¥500,000
10. 2001：Grant-in-Aid from the Ministry of Health,
Labor and Welfare of Japan（H-12-AIDS-004）
Studies on the Management of Complication in
HIV/AIDS” ¥1,005,000,000
11. Grant-in-Aid from the Ministry of Health, Labor andWelfare of Japan Socio-epidemiological Studies on
the Trends of HIV/AIDS in Japan and Its
Intervention” ¥500,000
12. 2002：Grant-in-Aid from the Ministry of Health,
Labor and Welfare of Japan（H-12-AIDS-004）
Studies on the Management of Complication in
HIV/AIDS” ¥63,000,000
13. Grant-in-Aid from the Ministry of Foreign Affairs ofJapan, US-Japan Cooperative Medical Science
Program AIDS Panel ¥6,500,000
14. Grant-in-Aid from the Ministry of Health, Labor andWelfare of Japan Socio-epidemiological Studies on
the Trends of HIV/AIDS in Japan and Its
Intervention” ¥500,000

Select Publications
1. S. Kimura, J. Matsuda, S. Ikematsu, K. Miyazono, A.
Ito, T. Nakahata, M. Minamitani, K. Shimada, Y.
Shiokawa, and F. Takaku; Efficacy of recombinanthuman granulocyte colony-stimulating factor on neutropenia in patients with AIDS. AIDS 4: 1251-1255,
1990
2. S. Oka, K. Urayama, Y. Hirabayashi, K. Ohnishi, H.
Goto, K. Mitamura, S. Kimura, and K. Shimada;
Quantitative estimation of human immunodeficiency
virus type-1 provirus in CD4+ T lymphocytes using
the polymerase chain reaction. Mol. and Cellular
Probes 5: 137-142, 1991
3. T. Toyoshima, S. Kimura, S. Muramatsu, H.
Takahagi, and K. Shimada; A sensitive nonisotopic
method for the determination of intracellular azidothymidine 5’-mono-, 5’-di-, and 5’-tri-phosphate.
Analyt. Biochem. 196: 302-307, 1991
4. K. Kitada, S. Oka, S. Kimura, K. Shimada, T.
Serikawa, J. Yamada, H. Tsunoo, K. Egawa, and Y.
Nakamura; Detection of Pneumocystis carinii
sequence by polymerase chain reaction: animal models and clinical application to noninvasive specimens.
J. Clin. Microbiol. 29: 1985-1990, 1991
5. S. Oka, K. Urayama, Y. Hirabayashi, S. Kimura, K.
Mitamura, and K. Shimada; Human immunodeficiency virus DNA copies as a virologic marker in a clinical trial with β-interferon. J. Acquir. Immun. Def.
Synd. 5: 707-711, 1992

6. S. Kimura; Current state of AIDS treatment - Can
AIDS be controlled? - . Asian Med. J. 35: 251-259,
1992
7. S. Kimura, S. Oka, T. Toyoshima, Y. Hirabayashi, Y.
Kikuchi, K. Mitamura, and K. Shimada; A randomized trial of reduced doses of azidothymidine inJapanese patients with human immunodeficiency
virus type 1 infection. Intern. Med. 31: 871-876,
1992
8. S. Oka, M. Goto, Y. Kaji, S. Kimura, K. Matsuda, Y.
Asai, M. Sanada, S. Nakagawa, M. Inoue, and K.
Shimada; Synergistic activity of imipenem/cilastatin
combined with cefotiam against methicillin-resistant
Stapylococcus aureus. J. Antimicrob. Chemother. 31:
533-541, 1993
9. K. Yamada, S. Kimura, M. Negishi, J. Takamatsu, M.
Inagaki, M. Aihara, Y. Nishida, K. Mori, K. Fukutake,
J. Mimaya, N. Takata, and K. Shimada. Phase I/II
trial of didanosine (2’, 3’-dideoxyinosine) in hemophiliac patients with AIDS or AIDS-related complex.
Clin. Diagnostic Virol. 1: 245-256, 1993
10. S. Oka, K. Kitada, T. Kohjin, Y. Nakamura, S.
Kimura, and K. Shimada; Direct monitoring as well
as sensitive diagnosis of Pneumocystis carinii pneumonia by polymerase chain reaction on sputum samples. Mol. Cellular Probes 7; 419-424, 1993
11. J. Hoy, E. J. Wright, and S. Kimura; Early HIV infection. AIDS 8 (suppl 2): s29-s35, 1994
12. T. Shioda, S. Oka, S. Ida, K. Nokihara, H. Toriyoshi,
S. Mori, Y. Takebe, S. Kimura, K. Shimada, and Y.
Nagai; A naturally occurring single basic amino acidsubstitution in the V3 region of HIV-1 Env protein
alters the viral cellular host range and antigenic
structure. J. Virol. 68: 7689-7696, 1994
13. H. Furukawa, K. Momota, T. Agatsuma, I.
Yamamoto, S. Kimura, and K. Shimada; Mechanismof inhibition of HIV-1 infection in vitro by guaninerich oligonucleotides modified at the 5’ terminal bydimethoxytrityl residue. Nucleic Acids Res. 22: 56215627, 1994
14. S. Watanabe, M. Shinkai, S. Hitomi, H. Kozuka, S.
Kimura, K. Shimada, R. Hondo, and N. Yamaguchi; Apolymorphic region of the human cytomegalovirus
genome encoding putative glycoproteins. Arch. Virol.
137: 117-121, 1994
15. A. Yasuoka, S. Oka, K. Komuro, H. Shimizu, K.
Kitada, S. Shibahara, T. Takeuchi, S. Kondo, K.
Shimada, and S. Kimura; Successful treatment with
Benanomicin A (ME1451) of Pneumocystis carinii
pneumonia in mice. Antimicrobial Agents
Chemother. 39: 720-724, 1995
16. A. Suzaki, M. Kimurua, S. Kimura, K. Shimada, M.
Miyaji, and L. Kaufman; An outbreak of histoplasmosis contacted by Japanese visitors to a bat-inhabited cave near Manaus, Brazil. J. Mycol. Med. 5: 40-43,
1995
17. M. Goto, S. Oka, N. Tachikawa, K. Kitada, M. Wada,
C. Abe, K. Shimada, and S. Kimura; KatG sequence
deletion is not the major cause of isoniazid resistancein Japanese and Yemei Mycobacterium tuberculosis

isolates. Molecular and Cellular Probes 9: 433-439,
1995
18. A. Yasuoka, N. Tachikawa, K. Kaoru, S. Kimura, andS. Oka; (1-3)β-D-glucan as a quantitative serologicalmarker for Pneumocystis carinii pneumonia. Clin.
Diag. Lab. Immunol. 3: 197-199, 1996
19. H. Hotoda, K. Momota, H. Furukawa, T. Nakamura,and M. Kaneko S. Kimura and K. Shimada;
Biologically active oligodeoxyribonucleotides- IV:
Anti-HIV activity of TGGGAG having hydrophobic
substituent at 5’-end via phosphodiester linkage.
Nucleosides & Nucleotides, 15: 531-538, 1996
20. H. Shiga, T. Shioda, H. Tomiyama, Y. Takamiya, S.
Oka, S. Kimura, Y. Yamaguchi, T. Gojoubori, H-G.
Rammensee, K. Miwa, and M. Takiguchi;
Identification of multiple HIV-1 cytotoxic T-cell epitopes presented by human leukocyte antigen B35
molecules. AIDS. 10: 1075-1083, 1996
21. S. Oka, Y. Nagata, Y. Fujino, A. Yasuoka, N.
Tachikawa, K. Shimada, S. Kimura, A. Iwamoto, M.
Mochizuki; CD8+ T lymphocytes in cytomegalovirus
retinitis in a patient with AIDS. Brit. J. Ophtalmol.
80: 1108-1109, 1996
22. S. Kimura; Neurologic manifestations in humanimmunodeficiency virus infection/ acquired immunodeficiency syndrome (Editorial): Intrenal Medicine
36: 160-161,1997
23. S. Ida, H. Gatanaga, T. Shioda, Y. Nagai, N.
Kobayashi, K. Shimada, S. Kimura, A. Iwamoto, andS. Oka; HIV type 1 V3 variation dynamics in vivo:
Long-term persistence of non-syncytium-inducing
genotypes and transient presence of syncytiuminducing genotypes during the course of progressive
AIDS. AIDS Research and Human Retroviruses 13:
1597-1609, 1997
24. K. Momota, H. Furukawa, S. Kimura, H. Hotoda, S.
Oka, and K. Shimada; Antiviral and biological properties of dimethoxytrityl-linked guanine-richoligodeoxynucleotides that inhibit replication by primary clinical human immunodeficiency virus type-1
isolates. Antiviral Chem. Chemother. : 8: 497-505,
1997
25. H. Yotsuyanagi, K. Yasuda, S. Iino, K. Moriya, Y.
Shintani, H. Fujie, T. Tsutsumi, S. Kimura, and K.
Koike; Persistent viremia after recovery from selflimited acute hepatitis B. Hepatology 27: 1377-1382,
1998
26. K. Moriya, H. Fujie, Y. Shimtani, H. Yotsuyanagi, T.
Tsutsumi, K. Ishibashi, Y. Matsuura, S. Kimura, T.
Miyamura and K. Koike; The core protein of hepatitis C virus induces hepatocellular carcinoma in transgenic mice. Nature Medicine, 4 (9): 1065-1067, 1998.
27. S. Hitomi, S. Baba, H. Yano, Y. Morisawa and S.
Kimura; Antimicrobial effects of electrolytic productsof sodium chloride - Comparative evaluation withsodium hypochlorite solution and efficacy in handwashing. J. Jpn. Assoc. Infect. Dis. 72 (11): 11761181, 1998
28. K. Koike, K. Moriya, H. Yotsuyanagi, Y. Shintani, H.
Fujie, T. Tsutsumi, and S. Kimura; Compensatory73apoptosis in preneoplastic liver of a transgenic
mouse model for viral hepatocarcinogenesis. Cancer
Lett 134 (2): 181-186, 1998
29. M. Koizumi, Y. Ozawa, R. Yagi, T. Nishigaki, M.
Kaneko, S. Oka, S. Kimura, A. Iwamoto, Y. Komatsu,
and E. Otsuka; Design and anti-HIV-1 activity ofhammerhead and hairpin ribozymes containing a stable loop. Nucleosides & Nucleotides 17 (1-3) : 207218, 1998
30. H. Kashiwase, K. Momota, T. Ohmine, T. Komai, T.
Kimura, T. Katube, T. Nishigaki, S. Kimura, K.
Shimada and H. Furukawa; A new fluoroquinolone
derivative exhibits inhibitory activity against humanimmunodeficiency virus type 1 replication.
Chemotherapy 45: 48-55, 1999
31. N. Tachikawa, M. Goto, Y. Hoshino, H. Gatanaga, A.
Yasuoka, T. Wakabayashi, Y. Katano, S. Kimura, S.
Oka, and A. Iwamoto; PCR detects Toxoplasma
gondii, Epstein-Barr virus, and JC virus DNAs in the
cerebrospinal fluid in AIDS patients with focal CNS
complications. Intern. Med. 38: 556-562, 1999
32. H. Fujie, H. Yotsuyanagi, K. Moriya, Y. Shintani, T.
Tsutsumi, T. Takayama, M. Makuuchi, Y. Matsuura,
T. Miyamura, S. Kimura, and K. Koike; Steatosis andintrahepatic hepatitis C virus in chronic hepatitis. J.
Med. Virol. 59: 141-145, 1999
33. Y. Shintani, H. Yotsuyanagi, K. Moriya, H. Fujie, T.
Tsutsumi, Y. Kanegae, S. Kimura, I. Saito, and K.
Koike; Induction of apoptosis upon the switch-on of
the hepatitis B virus X gene mediated by Cre/loxP
recombinase system. J. Gen. Virol. 180: 3257-3266,
1999
34. E. Noiri, T. Yokomizo, A. Nakao, T. Izumi, T. Fujita,
S. Kimura, and T. Shimizu; An in vivo approach
showing the chemotactic activity of leukotriene B4 inacute renal ischemic-reperfusion injury. Proc. Natl.
Acad. Sci. 97 (2): 823-828, 2000
35. S. Hashimoto, T. Matsumoto, M. Nagai, Y.
Matsuyama, Y. Nakamura, T. Umeda, M. Kamakura,
S. Ichikawa, S. Kimura, K. Fukutomi, and M. Kihara;
Delays and continuation of hospital visits among
HIV-infected persons and AIDS cases in Japan. J.
Epidemiol. 10 (1): 65-70, 2000
36. Y. Shintani, H. Yotsuyanagi, K. Moriya, H. Fujie, T.
Tsutsumi, T. Takayama, M. Makuuchi, S. Kimura,and K. Koike; The significance of hepatitis B virus
DNA detected in hepatocellular carcinoma of
patients with hepatitis C. Cancer 88 (11): 2479-2486,
2000
37. U. Machida, M. Kami, T. Fukui, Y. Kazuyama, M.
Kinoshita, Y. Tanaka, Y. Kanda, S. Ogawa, H. Honda,
S. Chiba, K. Mitani, Y. Muto, K. Osumi, S. Kimura,and H. Hirai; Real-time automated PCR for early diagnosis and monitoring of cytomegalovirus infection
after bone marrow transplantation. J. Clin. Microbiol.
38 (7): 2536-2542, 2000
38. A. Yoneyama, H. Yano, S. Hitomi, K. Okuzumi, R.
Suzuki, and S. Kimura; Ralstonia pickettii colonization of patients in an obstetric ward caused by a con-74

taminated irrigation system. J. Hosp. Infection 46:
77-80, 2000
39. S. Hitomi, M. Kubota, N. Mori, S. Baba, H. Yano, K.
Okuzumi, and S. Kimura; Control of a methicillinresistant Staphylococcus aureus outbreak in a neonatal intensive care unit by unselective use of nasal
mupirocin ointment. J. Hosp. Infection 46: 123-129,
2000
40. H. Yotsuyanagi, Y. Shintani, K. Moriya, H. Fujie, T.
Tsutsumi, T. Kato, K. Nishioka, T. Takayama, M.
Makuuchi, S. Iino, S. Kimura, and K. Koike; Virologic
analysis of non-B, non-C hepatocellular carcinoma inJapan: frequent involvement of hepatitis B virus. J.
Infect. Dis. 181: 1920-8, 2000
41. E. Noiri, A. Nakao, A. Oya, T. Fujita, and S. Kimura;
Hepatitis C virus in blood and dialysate in hemodialysis. Am. J. Kidney Dis. 37 (1): 38-42, 2001
42. H. Fujie, K. Moriya, Y. Shintani, H. Yotsuyanagi, S.
Iino, S. Kimura, and K. Koike; Hepatitis B virus genotypes and hepatocellular carcinoma in Japan.
Gastroenterology 120: 1564-1565, 2001
43. K. Tsuchiya, S. Matsuoka, A. Hachiya, A. Yasuoka,
N. Tachikawa, Y. Kikuchi, I. Genka, K. Teruya, S.
Kimura and S. Oka; Accumulation of lopinavir resistance-associated mutations over 3 years follow-up ofpatients on highly active antiretroviral therapy:
implication in salvage therapy. AIDS 15 (9): 11831184, 2001
44. H. Fujie, K. Moriya, Y. Shintani, T. Tsutsumi, T.
Takayama, M. Makuuchi, S. Kimura and K. Koike;
Frequent β-catenin aberration in human hepatocellular carcinoma. Hepatology Research 20: 39-51,
2001
45. K. Moriya, K. Nakagawa, T. Santa, Y. Shintani, H.
Fujie, H. Miyoshi, T. Tsutsumi, T. Miyazawa, K.
Ishibashi, T. Horie, K. Imai. T. Todoroki, S. Kimura
and K. Koike; Oxidative stress in the absence of
inflammation in a mouse model for hepatitis C virusassociated hepatocarcinogenesis. Cancer Res. 61:
4365-4370, 2001
46. Y. Yamaguchi, S. Fukuhara, T. Nagase, T. Tomita, S.
Hitomi, S. Kimura, H. Kurihara and Y. Ouchi; A
novel mouse β-defensin, mBD-6, predominantly
expressed in skeletal muscle. J. Biol. Chem. 276 (34):31510-31514, 2001
47. A. Yoshida, S. Hitomi, T. Fukui, H. Endo, Y.
Morisawa, Y. Kazuyama, K. Osumi, S. Oka and S.
Kimura; Diagnosis and monitoring of humancytomegalovirus diseases in patients with humanimmunodeficiency virus infection by use of a realtime PCR assay. Clinical Infectious Diseases 33: 17561761, 2001
48. M. Tanaka, C. Yasuoka, I. Genka, N. Tachikawa, Y.
Kikuchi, K. Teruya, A. Yasuoka, Y. Hirabayashi, S.
Kimura and S. Oka; Sustained cytomegalovirus-specific CD4+ T cell response associated with prevention of recurrence of cytomegalovirus retinitis without secondary prophylaxis after highly active antiretroviral therapy in patients with AIDS. AIDS Res.
Human Retroviruses 17 (18): 1749-1756, 2001

49. S. Ida, N. Tachikawa, A. Nakajima, M. Daikoku, M.
Yano, Y. Kikuchi, A. Yasuoka, S. Kimura and S. Oka;
Influence of human immunodeficiency virus type 1
infection on acute hepatitis A virus infection. Clinical
Infectious Diseases 34, 379-85, 2002
50. K. Koike, K. Moriya and S. Kimura; Role of hepatitis
C virus in the development of hepatocellular carcinoma: Transgenic approach to viral hepatocarcinogenesis. J. Gastroenterol. Hepatol. 17: 394-400, 2002

75

Department of
Immunology
Outline and Research Objectives
Research projects, currently being conducted in our Department, have stemmed fromthe original identification of two cytokine genes between the late 1970s and early 1980s,
namely, the genes encoding human fibroblast interferon (renamed thereafter as IFN-β) andinterleukin-2 (IL-2), and their molecular characterization. In fact, the molecular distinction
and consolidation of an enormous number of cytokine molecules, which are usually produced simultaneously at low levels in many cell types, have made it possible to study eachcytokine as a single molecule and to elucidate their intracellular signaling mechanisms aswell as their gene regulation mechanisms.
Given the fact that these cytokines are intimately involved in the regulation of immunityand oncogenesis, the above initial studies have led us to further extend the characterizationof these cytokine systems in the context of the regulation of immune responses and oncogenesis. More recently, we have also initiated a project linking the immune system andbone remodeling in the context of cytokine signaling cross talk. Thus, our current research
objectives are aimed at clarifying the following.
1. The mechanisms of signaling and transcription networks operating in the IFN-α/β system in innate immune responses.
2. The function and regulation of the transcription factor family, i.e., the interferon regulatory factors (IRFs), in innate and adaptive immune responses.
3. The mechanisms of regulation of antigen-presenting cells (APCs), typically dendrite cells(DCs), by IFNs and other cytokines and toll-like receptors (TLRs).
4. The regulation of ontogenesis by IFN-α/β and the tumor suppressor p53, particularly
their mutual cooperation and function on target genes.
5. The regulation of bone remodeling by RANKL of the TNF family member and its signaling cross talks with IFNs and other cytokines.
The above projects are being conducted in a coordinated manner with much emphasis
on interaction with researchers and students inside and outside of the Department, and collaborations, including international ones, are highly encouraged. The important objectives of our Department include the fostering of independent scientists with international
experience, and search for new concepts critical for future developments in immunology
and cancer biology.

Faculties and Students
Professor and Chair Tadatsugu Taniguchi, Ph.D. (1995~)
Lecturer
Akinori Takaoka, M.D., Ph.D. (2000~)
Associate .................................2
Postdoctoral Fellow ................4
Graduate student ..................12
Research student.....................1
Technical Assistant ................2
Secretary .................................2

Past Research and Major Accomplishments
T. Taniguchi, Professor and Chairman of thisDepartment, obtained his Ph. D. at the University of
Zurich in 1978 (Title of doctoral dissertation: RNARNA interactions in the process of 70S initiation com-76

plex formation between E. coli ribosomes and bacteriophage Qb RNA), and returned to work at The
Cancer Institute, Japanese Foundation for Cancer
Research in Tokyo, wherein the molecular characterization of soluble mediators of the immune response,
now collectively termed cytokines, was initiated. In
fact, we initially identified and characterized thegenes encoding IFN-β and IL-2, and established the
production of cytokines by recombinant DNA technology (Refs 1-6). The subsequent characterizationsof the promoters of these genes, performed at theinstitute, were further extended after we moved to
the Institute for Molecular and Cellular Biology ofOsaka University, and we identified distinct regulatory cis-elements within the promoter of IFN-β gene,which function as a virus-inducible enhancer (Refs. 7,
12). The work on the IFN-β enhancer led us to thediscovery of a novel family of transcription factors,
the IRF family, which now has nine members (Refs.
15, 18, 48). In fact, IFN-α/β genes as well as a number of IFN-inducible genes were shown to be involved
in regulating cell proliferation and to be potential critical target genes of IRFs. We also identified the regulatory sequences of the IL-2 gene, which have since
been extensively studied in the context of T cell activation and anergy by many other groups (Ref. 9). We
also continued working on the mechanisms of IL-2
signaling by characterizing and analyzing the structure and function of the IL-2 receptor complex. The
achievements, made during and after transition to thecurrent Department, are briefly summarized below oneach project.
(1) Gene regulation in IFN-α/β system; Operationof “revving up” mechanism for robust antiviral
responses.
The work on the IFN-β enhancer led us to discover
the IRF family of transcription factors. A number of
IFN-inducible genes were shown to be involved inregulating cell proliferation and to be potential criticaltarget genes of IRFs (Ref. 48). In fact, to evaluate thediverse biological effect of IRFs in vivo, we initiallyestablished the mice deficient in IRF-1 or IRF-2 genein collaboration with T. Mak and his colleagues,
University of Toronto (Ref. 29). We first found thatIRF-1 is a critical regulator for the antiviral response
by IFNs, but that it is not essential for virus-induced
IFN-α/β gene expression (Refs. 29, 31). More recently, we and others have focused on other two related
IRF members, IRF-3 and IRF-7. These two factors
reside in the cytoplasm of uninfected cells and undergo translocation to the nucleus after viral infection.
We adduced experimental evidence that these two
IRFs indeed activate the IFN promoters, and the exact
role of these factors became clear when we generated
mice deficient in these genes (Refs. 45, 48, 49). It was
found that cells have an acquired auto-amplification
mechanism for efficient IFN-α/β gene expression.
Briefly, viral infections first result in the phosphorylation of the constitutively expressed IRF-3, and thephosphorylated IRF-3 primarily activates the IFN-β
promoter. Once IFN-β is produced, it signals the cellto activate ISGF3, a heterotrimer complex consistingof STAT1, STAT2 and IRF-9; ISGF3 in turn induces
IRF-7 gene expression. The de novo produced IRF-7
then undergoes virus-induced phosphorylation, similar to IRF-3, and activates the IFN-α/β promoters.
Thus, massive IFN-α/β production can be achieved
through the positive-feedback loop (Refs. 48, 49).
We also found that the constitutive, IRF-3/IRF-7independent production of IFN-α/β in uninfected cells
is critical for setting the IRF-7 expression levels that
determine whether or not the above-mentioned posi-

tive-feedback mechanism will operate effectively upon
viral infection. We discussed the significance of spontaneous IFN-α/β gene expression, in the context ofwhat we proposed as the “revving up” model. In fact,a weak IFN-α/β signaling also provides a foundation
for cells to respond efficiently to other cytokines,
such as IFN-γ and IL-6 (Ref. 49). In the “revving up”
by the IFN-α/β signal, signaling molecules remain
constantly activated, albeit weakly, and the expression level of target genes such as IRF-7 (and probably
many others) is maintained, thereby providing a foundation for more efficient signaling, either in that pathway or in different pathways. Thus, the consumptionof cellular resources is not futile but a regulated“trade-off” to provide the cell with a greater dynamic
range (signal-to-noise) in its response to stimuli (Ref.
49). It is likely that mechanisms similar to what we
found in the context of the “revving up” model may
be operational in other immune responses, since theimmune system generally requires such mechanisms
so as to render the operation against invading
pathogens effective.
(2) Function of IRF-1 and IRF-2: Regulation of
immunity and oncogenesis.
In continuation of our analysis of the IRF-1-deficient mice, we found that IRF-1 is absolutely required
for Th1 type of CD4+ T cell responses and NK cell differentiation. In fact, IRF-1 is essential for the induction of IL-12 and inducible nitric oxide synthase,
which are critical for the induction and effector phaseof Th1 response, respectively, and it is also essential
for IL-15 gene expression in stroma cells to induce NK
cell development (Refs. 37, 38, 48). In addition, IRF-1
is found to be a tumor susceptibility gene. IRF-1 isrequired in the induction of apoptosis of oncogeneexpressing fibroblasts and DNA-damaged, proliferating mature T cells (Refs. 35, 36, 42). In DNA-damaged fibroblasts, IRF-1 cooperates with p53 to induce
cell cycle arrest by activating the p21WAF1/CIP1 gene. We
have shown that the loss of IRF-1 alleles per se has noeffect on spontaneous tumor development in themouse, but that it dramatically exacerbates previous
tumor predispositions caused by the c-Ha-ras transgene or nullizygosity for p53. In addition, notable
alterations in the tumor spectrum of IRF-1, p53 double-deficient mice indicated that IRF-1 is not hypostatic to p53. We also adduced evidence that the loss-offunction mutation in the IRF-1 gene may be involvedin cancer development in humans (Ref. 42, 48). In
continuation of this project, we attempted to isolate
genes functioning downstream of IRF-1/p53, andidentified a novel gene, Noxa . The Noxa geneencodes a BH3-only protein of the Bcl-2 family, and its
expression is actually dependent on p53; it also
induces apoptosis when it is overexpressed (Ref. 43).

77

Although IRF-2 was originally identified as a negative regulator of IRF-1, it has a unique function in negatively regulating IFN-α/β signaling. IRF-2 is a very
stable nuclear factor that attenuates transcription by
the IFN-activated ISGF3. Interestingly, IRF-2-deficient
mice spontaneously develop an inflammatory skindisease, resembling psoriasis. CD8+ T cells appear to
be involved in the pathogenesis of this condition, andCD8+ T cells from these mice are hyper-responsive toantigen stimulation in vitro, which is accompanied bya markedly upregulated expression of genes induced
by IFN-α/β (Refs.46, 48). Thus, IRF-2 is necessary forbalancing the beneficial and harmful effects of the
spontaneous IFN-α/β signaling, as described above.
(3) Characterization and functional studies on the
IL-2 receptor complex.
The use of recombinant IL-2 made it possible tostudy this “T cell growth factor” in the context of regulation of intracellular signaling mechanisms. T.
Waldmann and colleagues first discovered the IL-2
receptor, as an IL-2Rα chain, and it turned out thatthe functional receptor consists of subunits (Refs. 28,
34). We then isolated the second chain, IL-2Rβ chain,and showed for the first time that this cytokine receptor, lacking any catalytic domains, can transmit thesignals to the cell interior by recruiting non-receptor
tyrosine kinases (Refs. 8, 10, 17, 19, 34). In addition,
it was also shown that multiple signaling molecules
are recruited to the receptor, which play distinct functions. It is now known that IL-2R consists of IL-2Rα,β (termed cβ) and γ (termed cγ) chains.
We also took an in vivo approach to study the distinct domains of the IL-2Rβ cytoplasmic region byintroducing each mutant cDNA that lacked one ofthese domains into IL-2Rβ-deficient mice. We showedthat the lack of the membrane-distal H-region, whichmediates activation of STAT5/STAT3 transcription
factors, selectively affects the development of naturalkiller (NK) cells and T cells bearing γδ T cell receptors.
In contrast, the A-region, which is located next to the
H-region and mediates the activation of Src-family
protein tyrosine kinases, contributes to the downregulation of the T cell proliferation function. The IL2Rβc null mutant mice develop severe autoimmune
symptoms, but these are all suppressed following theexpression of either of the mutant cDNAs (Ref. 40).
(4) Osteoimmunology: Signaling cross talks
between IFNs and RANKL in bone remodeling.
The regulation of osteoclast differentiation is an
aspect central to the understanding of the pathogenesis and the treatment of bone diseases such as
autoimmune arthritis and osteoporosis. In fact,
excessive signaling by RANKL, a TNF family member
essential for osteoclastogenesis, may contribute to78

such pathological conditions. We found unique signaling cross talks between RANKL and IFNs. We provided evidence that activated T cells maintain bonehomeostasis by counterbalancing the action of
RANKL through IFN-γ production. IFN-γ induces the
rapid degradation of the RANK adapter protein,
TRAF6, resulting in the strong inhibition of theRANKL-induced activation of NF-κB and JNK (Ref.
47). This work was introduced as a new area of
research called osteoimmunology [Arron, J. R. et al.
Nature, 408, 535, 2000]. More recently, we also foundthat RANKL induces the IFN-β gene, but not IFN-α
genes, in osteoclast precursor cells, and that IFN-β
strongly inhibits osteoclast differentiation by interfering with the RANKL-induced c-Fos expression (Ref.
50). The series of in vivo experiments revealed thatthese two IFN-mediated regulatory mechanisms, distinct with one another, are both important to keepbalancing the homeostatic bone resorption.
Collectively, these studies revealed novel aspects ofthe two types of IFNs, beyond their original roles in
immune response, and may offer a molecular basis forthe treatment of bone diseases.

Current Research
As described above, we have gained new insight
into the mechanisms of signaling and transcription
networks operating in the IFN-α/β system in immune
responses. The current research projects in our laboratory are summarized as follows:
1. Function and regulation of IRFs.
Still elusive is the mechanism(s) by which IRF-3
and IRF-7 are activated by viruses. We have been trying to identify a virus-activated kinase(s), which is
responsible for this activation by taking a proteomics
approach. We have identified several molecules thatare associated with IRF-3, and these molecules are
currently subjected to functional analyses to determine their involvement in IRF-3 regulation. We havegenerated IRF-7-deficent mice and are now confirming
the role of IRF-7 in IFN-α/β gene induction in distinct
cell populations, particularly, in bone marrow-derived
cells, such as DCs. Our preliminary data indicate thatgerminal center formation is impaired in the mutant
mice, and we are going to analyze the underlying
mechanisms. The generation of IRF-3/IRF-7 double
deficient mice is also under way. As for IRF-1, we
have been working on the role of IRF-1 in CD4+ T celldifferentiation, particularly the identification of the
IRF-1 target genes.
2. Mechanisms of regulation of antigen-presenting cells (APCs), typically dendritic cells (DCs).
We found that the IFN-α/β system is criticallyinvolved in DC maturation, which is induced by TLRs.
We are currently studying the mechanism by which
TLR expression is regulated by the IFN-α/β system.
We are also working on the differentiation and maturation of DCs, which are found to be abnormal in IRF2-deficient mice.
3. Regulation of oncogenesis by IFN-α/β and thetumor suppressor p53.
A critical function of p53 in tumor suppression is
the induction of apoptosis in cells exposed to noxious
stresses, such as DNA damage. We have previously
shown that Noxa belongs to the BH3-only subfamilyof the Bcl-2 family proteins and that Noxa overexpression causes apoptosis of some cells. More recently,
we generated mice deficient in the Noxa gene or inboth Noxa and Bax genes. In addition, we are continuously studying on the cooperation between IFN signaling and p53 pathways in relation to oncogenesis
using mutant mice that are defective in some of thesepathways.
4. Regulation of osteoclast differentiation.
During the differentiation process of hematopoietic stem cells to osteoclasts with bone-resorbing activity, transcription factors such as PU.1, c-Fos, NF-κB,
and MITF play a critical and specific role. To date,however, little is known about how RANKL, but notother cytokines that activate similar pathways, specifically induces the terminal differentiation of osteoclasts through a specific transcriptional program.
Previous observations suggest that RANKL signaling
activates an as yet unknown pathway(s) that specifically invokes the transcriptional program leading thecells to undergo terminal differentiation. To gain
insight into the mechanism of the RANKL-specificinduction of the osteoclast differentiation program,
we took a genome-wide screening approach to identify genes specifically induced by RANKL, but not by
IL-1, and came up with the NFATc1 transcription factor. We are currently analyzing the complex signaling
and transcription networks in the RANKL-cFos/TRAF6-NFATc1 pathways to gain more insight
into the differentiation programs.

Future Prospects
From the inception of the molecular analyses of
the cytokine systems in the late 1970s, research projects in our laboratory have progressed and differentiated in many ways. This was natural and importantfor our laboratory, particularly in fostering the nextgeneration scientists who are independent and haveinternational experience. In fact, more than ten former colleagues have been promoted to full professor
in renowned universities. The discovery and func-

tional analyses of the IRF family of transcription factors provided new insight into the molecular mechanisms underlying the efficient induction of IFN-α/β
genes, as well as the regulation of immune responses
and oncogenesis. As for future prospects, it is important to elucidate the mechanisms of activation ofthese transcription factors in virus-infected cells.
Considering the increasing interest in the IFN-α/βsystem induction by nonviral pathogens in antigen
presenting cells, it will also be important to elucidate
the role of IFNs and IRFs in linking the innate andadaptive immune systems. We are beginning to see a
new link between IFN systems and p53, and this will
hopefully reveal a new and interesting area of
research in cancer biology. We also continue elucidating the RANKL-mediated mechanisms of bone remodeling. We hope that our studies will provide novel
therapeutic strategies for controlling infectious andneoplastic diseases, as well as bone diseases. In fact,
it is also our long-term goal to gain insight into themechanisms of autoimmunity and tumor immunity,
so as to provide a means for the treatment of these
diseases.

Research Grants
1. A grant for Advanced Research on Cancer from the
Ministry of Education, Culture, Sports, Science, andTechnology of Japan (1998-to date)
2. The research for the Future Program (96L0037) fromJapan Society for the Promotion of Sciences (19951999).
3. Human Frontier Science Program (1996-1998); In collaboration with Drs. D. Littman (USA), B. Malissen
and O. Acuto (France)

Selected Papers (50)
1. Taniguchi, T., Sakai, M., Fujii-Kuriyama, Y.,
Muramatsu, M., Kobayashi, S.and Sudo,T. ;Construction and identification of a bacterial plasmid
containing the human fibroblast interferon genesequence. Proc. Jpn. Acad., 55B, 464-469, 1979.
2. Taniguchi, T., Ohno, S., Fujii-Kuriyama, Y. and
Muramatsu, M. ; The nucleotide sequence of humanfibloblast interferon cDNA. Gene, 10, 11-15, 1980.
3. Taniguchi, T., Mantei, N., Schwarzstain, M., Nagata,
S., Muramatsu, M. and Weissmann, C. ; Humanleukocyte and fibroblast interferons are structurally
related. Nature, 285, 547-549, 1980.
4. Ohno, S. and Taniguchi, T. ; Structure of a chromosomal gene for human interferon-β. Proc. Natl. Acad.
Sci. USA, 78, 5305-5309, 1981.
5. Taniguchi, T., Matsui, H., Fujita, T., Takaoka, C.,
Kashima, N., Yoshimoto, R. and Hamuro, J. ;Structure and expression of a cloned cDNA for
human interleukin-2. Nature, 302, 305-310, 1983.

79

6. Kashima, N., Nishi-Takaoka, C., Fujita, T., Taki, S.,
Yamada, G., Hamuro, J. and Taniguchi,T. ; Uniquestructure of murine interleukin-2 as deduced fromcloned cDNAs. Nature, 313, 402-404, 1985.
7. Fujita, T., Ohno, S., Yasumitsu, H. and Taniguchi, T. ;Delimitation and properties of DNA sequences
required for the regulated expression of human interferon-β gene. Cell, 41, 489-496, 1985.
8. Hatakeyama, M., Minamoto, S., Uchiyama, T., Hardy,
R. R., Yamada, G. and Taniguchi, T. ; Reconstitutionof functional receptor for human interleukin-2 inmouse cells. Nature, 318, 467-470, 1985.
9. Fujita, T., Shibuya, H., Ohashi, T. Yamanishi, K. andTaniguchi, T. ; Regulation of human interleukin-2
gene. : Functional DNA sequences in the 5’ flanking
region for the gene expression in activated T lymphocytes. Cell, 46, 401-407, 1986.
10. Taniguchi, T., Fujita ,T., Hatakeyama, M., Mori, H.,
Matsui, H., Sato, T., Hamuro, J., Minamoto, S.,
Yamada, G. and Shibuya, H. ; Interleukin-2 and its
receptor: Structure and functional expression of thegenes. Cold Spring Harbor Symp., LI, 577-586, 1986.
11. Maruyama, M., Shibuya, H., Harada, H.,
Hatakeyama, M., Seiki, M., Fujita, T., Inoue, J-I.,
Yoshida, M. and Taniguchi, T. ; Evidence for aberrant activation of the interleukin-2 autocrine loop by
HTLV-1-encoded p40x and T3/Ti complex triggering.
Cell, 48, 343-350, 1987.
12. Fujita, T., Shibuya, H., Hotta, H., Yamanishi, K. andTaniguchi, T. ; Interferon-β gene regulation:
Tandemly repeated sequences of a synthetic 6bp
oligomer function as a virus inducible enhancer. Cell,
49, 357-367, 1987.
13. Yamada, G., Kitamura, Y., Sonoda ,H., Harada, H.,
Taki, S., Mulligan, R.C., Osawa, H., Diamantstein, T.,
Yokoyama, S. and Taniguchi, T. ; Retroviral expression of the human IL-2 gene in a murine T cell line
results in cell growth autonomy and tumorigenicity.
EMBO J., 6, 2705-2709, 1987.
14. Taniguchi, T. ; Regulation of cytokine gene expression. Annu. Rev. Immunol., 6, 439-464, 1988.
15. Miyamoto, M.,Fujita, T., Kimura, Y., Maruyama, M.,
Harada, H., Sudo, Y., Miyata, T. and Taniguchi, T. ;Regulated expression of a gene encoding a nuclear
factor, IRF-1, that specifically binds to IFN-β gene
regulatory elements. Cell, 54, 903-913, 1988.
16. Fujita, T., Kimura, Y., Miyamoto, M., Barsoumian, E.
L. and Taniguchi, T. ; Induction of endogenous IFNαand IFN-βgene by a regulatory transcription factor, IRF-1. Nature, 337, 270-272, 1989.
17. Hatakeyama, M., Tsudo,M., Minamoto, S., Kono, T.,
Doi, T., Miyata, T., Miyasaka, M. and Taniguchi, T. ;Interleukin-2 receptorβchain gene: Generation of
three receptor forms by cloned human αandβchain
cDNA’s. Science, 244, 551-556, 1989.
18. Harada, H., Fujita, T., Miyamoto, M., Kimura, Y.,
Maruyama, M., Furia, A., Miyata, T. and Taniguchi,
T. ; Structurally similar but functionally distinct factors, IRF-1 and IRF-2, bind to the same regulatory

80

elements of IFN and IFN-inducible genes. Cell, 58,
729-739, 1989.
19. Hatakeyama, M., Mori, H., Doi, T. and Taniguchi, T.
; A restricted cytoplasmic region of IL-2 receptorβ
chain is essential for growth signal transduction but
not for ligand binding and internalization. Cell, 59,
837-845, 1989.
20. Taniguchi, T., Hatakeyama, M., Minamoto, S., Kono,T., Doi, T., Tsudo, M., Miyasaka, M. and Miyata, T. ;Interleukin-2 receptorβchain: Molecular cloning andfunctional expression of the human cDNA. Cold
Spring Harbor Symposia on Quantitative Biology,
Vol. LIV, 689-694, 1989.
21. Harada, H., Willison, K., Sakakibara, J., Miyamoto,M., Fujita, T. and Taniguchi, T. ; Absence of the type
I IFN system in EC cells: Transcriptional activator(IRF-1) and repressor (IRF-2) genes are developmentally regulated. Cell, 63, 303-312, 1990.
22. Hatakeyama, M., Kono, T., Kobayashi, N., Kawahara,A., Levin, S. D., Perlmutter, R. M., and Taniguchi, T.
; Interaction of the IL-2 receptor with the src-family
kinase p56lck: Identification of novel intermolecularassociation. Science, 252, 1523-1528, 1991.
23. Shibuya, H., Yoneyama, M., Ninomiya-Tsuji, J.,
Matsumoto, K., and Taniguchi, T. ; IL-2 and EGF
receptors stimulate the hematopoietic cell cycle via
different signaling pathways: Demonstration of anovel role for c-myc. Cell, 70, 57-67, 1992.
24. Minami, Y., Kono, T., Yamada, K., Kobayashi, N.,
Kawahara, A., Perlmutter, R. M., and Taniguchi, T. ;Association of p56lck with IL-2 receptor β chain is
critical for the IL-2 induced activation of p56lck.
EMBO J., 12. 759-768, 1993.
25. Harada H., Kitagawa, M., Tanaka,N., Yamamoto, H.,
Harada, K., Ishihara, M., and Taniguchi, T. ; Antioncogenic and oncogenic potentials of interferon regulatory factors-1 and-2. Science, 259, 971-974, 1993.
26. Willman, C. L., Sever C. E., Pallavicini, M. G., Harada,
H., Tanaka, N., Slovak, M. L., Yamamoto, H.,
Harada, K., Meeker, T. C., List, A. F., and Taniguchi,
T. ; Deletion of IRF 1 , mapping to chromosome
5q31.1, in human leukemia and preleukemic
myelodysplasia. Science, 259, 968-971, 1993.
27. Minami, Y., Kono, T., Miyazaki, T., and Taniguchi, T.
; The IL-2 receptor complex: Its structure, function,and target genes. Annu. Rev. Immunol., 11, 245-267,
1993.
28. Taniguchi, T., and Minami, Y. ; The IL-2/IL-2 receptor system: A current overview. Cell, 73, 5-8, 1993.
29. Matsuyama, T., Kimura, T., Kitagawa, M., Pfeffer, K.,
Kawakami, T., Watanabe, N., K dig, M. T.,
Amakawa, R., Kishihara, K., Wakaeham, A., Potter,J., Furlonger, L. C., Narendran, A., Suzuki, H.,
Ohashi, P. S., Paige, J. C., Taniguchi, T. and Mak, T.
W.; Targeted disruption of IRF-1 or IRF-2 results inabnormal type I IFN gene induction and aberrant
lymphocyte development. Cell, 75, 83-97, 1993.
30. Tanaka, N., Ishihara, M., Kitagawa, M., Harada, H.,
Kimura, T., Matsuyama, T.,Lamphier, M. S., Aizawa,S., Mak, T. W. and Taniguchi, T.; Cellular commit-

ment to oncogene-induced transformation or apoptosis is dependent on the transcription factor IRF 1.
Cell, 77, 829-839, 1994.
31. Kimura, T., Nakayama, K., Penninger, J., Kitagawa,
M., Harada, H., Matsuyama, T., Tanaka, N., Kamijo,
R., Vilcek, J., Mak, T. W., Taniguchi, T. ;Involvement of the IRF-1 transcription factor in
antiviral responses to interferons. Science, 264, 19211924, 1994.
32. Miyazaki, T., Kawahara, A., Fujii, H., Nakagawa, Y.,
Minami, Y., Liu, Z-J., Oishi, I., Silvennoinen, O.,
Witthuhn, B.A., Ihle, J.N., and Taniguchi, T.;
Functional activation of Jak 1 and Jak3 by selective
association with IL-2 receptor subunits. Science, 266,
1045-1047, 1994.
33. Minami, Y., Nakagawa, Y., Kawahara, A., Miyazaki,T., Sada, K., Yamamura, H. and Taniguchi, T. ;Protein tyrosine kinase Syk is associated with andactivated by the IL-2 receptor, possible link with thec-myc induction pathway. Immunity , 2, 89-100,
1995.
34. Taniguchi, T. ; Cytokine signaling through nonreceptor protein tyrosine kinases. Science, 268, 251-255,
1995.
35. Tamura, T., Ishihara, M., Lamphire, M.S., Tanaka,N., Oishi, I., Aizawa, S., Matsuyama, T., Mak, T. W.,
Taki, S. and Taniguchi, T. ; An IRF-1-dependent
pathway of DNA damage-induced apoptosis in mitogen-activated T lymphosytes. Nature, 376, 596-599,
1995.
36. Tanaka, N., Ishihara, M., Lamphier, M., Nozawa, H.,
Matsuyama, T., Mak, T.W., Aizawa, S., Tokino, T.,
Oren, M. and Taniguchi, T. ; Cooperation of thetumor suppressors IRF-1 and p53 in response to
DNA damage. Nature, 382, 816-818, 1996.
37. Taki, S., Sato, T., Ogasawara, K., Fukuda, T., Sato,
M., Hida, S., Suzuki, G.,Mitsuyama, M., Shin, E.-H.,
Kojima, S., Taniguchi, T. and Asano, Y. ; Multistage
regulation of Th1-type immune responses by thetranscription factor IRF-1. Immunity, 6, 673-679,
1997.
38. Ogasawara, K., Hida, S., Azimi, N., Tagaya, Y., Sato,T., Yokochi-Fukuda, T.,Waldmann, T.A., Taniguchi,
T. and Taki, S. ; Requirement of IRF-1 for the
microenvironment supporting natural killer cell
development. Nature, 391, 700-703, 1998.
39. Miyazaki, T., Takaoka, A., Noguerira, L., Dikic, I.,
Fujii, H., Tsujino, S., Mitani, Y., Maeda, M.,
Schlessinger, J. and Taniguchi, T. ; Pyk2 is a downstream mediator of the IL-2 receptor-coupled Jak-signaling pathway. Genes. Dev. 12, 770-775, 1998.
40. Fujii, H., Ogasawara, K., Otsuka, H., Suzuki, M,
Yamamura, K., Yokochi, T, Miyazaki, T., Suzuki, H.,
Mak, T. W., Taki, S. and Taniguchi, T.; Functional
dissection of the cytoplasmic subregions of the IL-2
receptor βc chain in primary lymphocyte populations. EMBO J., 17, 6551-6557, 1998.
41. Takaoka, A., Tanaka, N., Mitani, Y., Miyazaki, T.,
Fujii, H., Sato, M., Kovarik, P,. Decker, T.,
Schlessinger, J. and Taniguchi, T.; Protein tyrosine

kinase Pyk2 mediates the Jak-dependent activationof MAPK and Stat1 in the IFN-γ, but not IFN-α,signaling. EMBO. J., 18, 2480-2488, 1999.
42. Nozawa, H., Oda, E., Nakao, K., Ishihara, M., Ueda,
S., Yokochi, T., Ogasawara, K., Nakatsuru, Y.,
Shimizu, S., Ohira, Y., Hioki, K., Aizawa, S.,
Ishikawa, T., Katsuki, M., Muto, T., Taniguchi, T.,
and Tanaka, N.; Loss of transcription factor IRF-1
affects tumor susceptibility in mice carrying the Haras transgene or nullizygosity for p53. Genes. Dev.
12, 1240-1245, 1999.
43. Oda, E., Ohki, R., Murasawa, H., Nemoto, J., Shibue,T., Yamashita,T., Tokino, T., Taniguchi, T. andTanaka, N.; Noxa, a BH3-only member of the Bcl-2
family, and candidate mediator of p53-induced apoptosis. Science, 288, 1053-1058, 2000.
44. Takaoka, A., Mitani, Y., Suemori, H., Sato, S.,
Yokochi, T., Noguchi, S., Tanaka, N., and Taniguchi,T.; Crosstalk between interferon-γand-α/βsignaling components at caveolar membrane domain.
Science, 288, 2357-2360, 2000.
45. Sato, M., Suemori, H., Hata, N., Asagiri, M.,
Ogasawara, K., Nakao, K., Nakaya, T., Katsuki, M.,
Noguchi, S., Tanaka, N., and Taniguchi, T.;Distinct
and essential roles of transcription factors IRF-3 andIRF-7 in response to viruses for IFN-α/βgene induction. Immunity, 13, 539-548, 2000.
46. Hida, S., Ogasawara, K., Sato, K., Abe, M.,
Takayanagi, H., Yokochi, T., Sato, T., Hirose, S.,
Shirai, T., Taki, S. and Tanicuhi, T.;CD8+ T cellmediated skin disease in mice lacking IRF-2, thetranscriptional attenuator of interferon-α/βsignaling. Immunity,13, 643-655, 2000.
47.Takayanagi, H., Ogasawara, K., Hida, S., Chiba, T.,
Murata, S., Sato, K., Takaoka, A., Yokochi, T., Oda,
H., Tanaka, K., Nakamura, k. and Taniguchi, T.; Tcell-mediated regulation of osteoclastogenesis by signalling cross-talk between RANKL and IFN-γ.
Nature, 408, 600-605, 2000.
48. Taniguchi, T., Ogasawara, K., Takaoka, A, andTanaka, N.; IRF family of transcription factors as regulators of host defense. Ann. Rev. Immunol. 19,
623-655, 2001.
49 Taniguchi, T. and Takaoka, A. ; A weak signal forstrong responses: Interferon -α/β Revisited.
Nature Rev. Mol. Cell Biology, 2, 378-386, 2001.
50. Takayanagi, H., Kim, S., Matsuo, K., Suzuki, H.,
Suzuki, T., Sato, K., Yokochi, T., Oda, H., Nakamura,K., Ida, N., Wagner, E. F., and Taniguchi, T.; RANKL
maintains bone homeostasis through c-Fos-dependent induction of interferon-β. Nature, 416, 744-749,
2002.

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Department ofRadiology
Outline and Research Objectives
Department of Radiology was established in 1932. Our department covers three major
fields that are, diagnostic radiology (imaging and intervention), radiation oncology (radiotherapy) and nuclear medicine. Research activities of radiology are being carried out in cooperation with department of clinical radiology of the University of Tokyo Hospital and withdepartment of radiology of the institute of medical science (IMS), the University of Tokyo.

Research Objectives
Diagnostic Radiology
To develop new techniques in data acquisition of CT and MRI
To clarify pathologic background of radiological findings.
To develop new system for interventional procedures.
Radiation oncology
To develop high precision radiotherapy system
To improve radiotherapy planning.
To clarify biological mechanisms of radiotherapy from molecular viewpoints.
Nuclear Medicine
To develop new techniques and radiotracers in functional imaging.
To improve image quality and quantitative accuracy in radiotracer imaging

Faculties and Students
Professor and Chair
Kuni Ohtomo MD (diagnostic radiology 1998-)
Associate Professors
Manabu Minami MD (diagnostic radiology 1999-)
Shigeki Aoki MD (diagnostic radiology 2000-)
Keiichi Nakagawa MD (radiation oncology 2002-)
Kohki Yoshikawa MD (diagnostic radiology 1997-) (IMS)
Lecturers
Toshimitsu Momose MD (nuclear medicine 1999-)
Osamu Abe MD (diagnostic radiology 2000-)
Naoto Hayashi MD (diagnostic radiology 2000-)
Masao Tago MD (radiation oncology 2001-)
Yoshitaka Masutani ED (computer science 2002-)
Yusuke Inoue MD (nuclear medicine 1999-) (IMS)
Associates ...............................9
Postdoctoral Fellow ................1
Graduate Students ................14
Residents.................................6
Secretaries ...............................982

Past Research and Major Accomplishments
Diagnostic Radiology
1. Visualization of cerebral arteries by contrastenhanced CT and MRI with special attention to separate between the lumen and the wall. (select publications 1-7,12)
We have revealed that dynamic contrast enhanced
CT and MRI can be used to visualize vascular lumen
as well as its wall.
We developed 3D-CT angiography for cerebral
arteries to visualize lumen of the arteries andaneurysms. We developed a new MR angiographic
technique, named “MRDSA” which can show the
hemodynamics with a frame rate of subsecond. We
also established the technique to see the wall of theintracranial arteries, using contrast-enhanced MRwith suppression of the luminal signal by presaturation pulse. We used this technique to see the irradiated intracranial arterial wall, which sometimes show
marked enhancement. We then established the way
to see the dynamic contrast changes of the wall itself.
In addition to diagnosis, we performed fundamental studies of MR-guided intravascular procedures.

2. Temporal Changes of the Apparent Diffusion
Coefficients (ADCs) of Water and Metabolites in Rats
with Hemispheric Infarction. (select publication 15)
The purpose was to clarify the temporal changes of
ADCs of cerebral metabolites during early focal ischemia.
in order to assess the pathophysiology of the reduction
in diffusion properties observed both in the ischemic
hemisphere and in the contralateral hemisphere.
3. Fully-Automated Segmentation of Colonic Walls
and Pulmonary Vessels (select publications 19-20)
We developed new segmentation methods, which
are applicable in both visualization of the colon andcomputer-aided diagnosis in the detection of polyps in
CT colonoscopy. A new method for automated segmentation of the pulmonary vascular tree in spiral CT
angiography was developed based on 3D image analysis techniques and anatomic knowledge. The results
are able to be applied in detection of pulmonary
embolism by limiting search area of thrombi.
4. Radiologic –pathologic correlation of various diseases (select publications 8-1116.18.21-26)
In the study the images with high spatial and contrast resolution are compared with the pathologic
findings and this kind of information is helpful fordifferential diagnosis of diseases. This approach has
been successful in the field of the CNS, liver, andovaries.
5. New virtual CT endoscopy software
To overcome some of the disadvantages of virtual
endoscopy, new software called computed sectional
probe (CSP) method has been developed. This software can be applied to the diagnosis of the gastrointestinal tract and tracheobronchial tree with improved
detectability and more accurate tumor staging.

beam placement in the high precision radiation therapy.

Nuclear Medicine
1. Brain activation studies by positron emission
tomography (PET) (select publications 39.40)
We investigated the following issues; (I) reproducibility of regional cerebral blood flow of restingstate measured by H2O-PET;(II) effect of physiologicalstimulation on cerebral blood flow;(III) identificationof activation areas on anatomical structures. Our datasuggest that relative rCBF images and their paired
subtraction are more suitable for tapping functional
localization. The changes of rCBF in association cortex
for more complicated tasks are, in general, smaller
than those in primary cortex. PET images were taken
in two different conditions: blindfolded resting state
and anti-saccade. We applied these activation techniques using H2O and PET to exploring motor function and language processing of Japanese language.
We investigated functional neuroanatomy of chewing.
We investigated Japanese language processing in
the brain using H2O-PET. The results suggested that
Kanji and Kana are processed differently.
2. Evaluation of the prognostic value of FDG-PET forrectal cancer (select publications 41)
We compared several variations of a semi-quantification method, the Standardized Uptake Values (SUV)
and to determine the most appropriate parameter forthe prognostic prediction and to propose the quantitative guideline of the FDG-PET. SUV2 was considered
to be a good prognostic indicator for long-term prognosis of rectal cancer patients.

Current Research
Radiation Oncology
1. Development of a comprehensive system for precise radiation therapy (select publications 27.30.)
A total system for radiation therapy composed of alinear accelerator unit with a microcomputer-controlled multileaf collimator and a CT scanner installedin the same treatment room was developed 17 years
ago. It was a prototype system of the widely prevailing high precision radiotherapy system.
2. Megavoltage CT scanning with a medical accelerator (select publication 29 32)
Megavoltage CT scanner using 4 MV and 6 MV
radiotherapy beam was developed. A detector system
is composed of 120 pairs of Cadmium Tungstate scintillators. Although spatial resolution is limited, this system has been used to verify positioning errors andDiagnostic Radiology
1. Evaluation and treatment of cerebrovascular diseases with multimodality (CT, MR, US, angiography);especially with X-ray angio-open MR system.
2. MR Diffusion Tensor Analysis and Visualization(select publications 13.14)
A new display method for tractography in diffusion tensor imaging was developed. Local reliability ofdiffusion tracking is evaluated based on anisotropy,
and paths of tracking are displayed in colors andopacity according to tracking reliability. By using several clinical data sets of volunteers and patients, themethod was validated.

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3. Further clinical application of 3D data acquired byMultidetector low CT (MDCT).
3-1. 3D display of secondary lobules of the lung
MDCT can generate contiguous thin section(1.25mm) CT images of the lung at every 0.6mm
under single-breath holding. 3-D data of lungparenchyma is abstracted and erosion of several pixels from the lung surface can be accomplished. The
structure of the secondary pulmonary lobule is clearly
seen. This information will be helpful to analyze diffuse lung diseases.
3-2. 3D display of the surface of the liver
Using multi-phase dynamic study of the liver,characteristics of enhancement in each pixel can be
abstracted. If the characteristics of enhancement of
the liver parenchyma are set on the time-density
curve, pixels showing similar enhancement pattern
will be differentiated automatically on computer. This
will be utilized to define the area of the liver
parenchyma and eventually the data of the area willshow the appearance of the liver surface three-dimensionally. The appearance will be almost the same as
the liver is observed by laparoscopy or at surgery.
This computer-aided automatic process will be useful
the overall evaluation of changes of the liver shapeand its deformity (atrophy and hypertrophy).
Radiation Oncology
1. Molecular analysis on reproductive tissue (selectpublications 34.35.38)
Radiation damage and recovery from ionizing radiation are being investigated from the molecular viewpoint.
2. Development of a C-arm mounted accelerator andof Dynamic Conical Conformal Radiation therapy(Dyconic CRT)
A C-arm mounted accelerator was developed. The
linac head was designed to move along the C-arm
with a maximum angle of 60 degrees (from a vertical
position toward the gantry). Simultaneous rotation of
the gantry creates a dynamic conical irradiation technique. Dyconic CRT was developed by combining thetechnique with continuous motion of multi-leaf collimator (MLC). Dyconic CRT enabled the precise delivery of non-coplanar beams without rotating the table.
Nuclear Medicine
1. Comparative studies of myocardial sympathetic
nerve function and striatal dopaminergic function in
Parkinson’s disease and its related disorders.
In all patients with parkinsonism, striatum to cerebellar (ST/CBL)Ratios were significantly reduced in
FDOPA-PET. Among them, only in PD and DLBD

84

patients, heart to mediastinum (H/M) ratios, index of
myocardial sympathetic nerve function, were significantly reduced in I-123 MIBG scans. Now we are accumulating the number of patients and try to analyze
the data of larger series of parkinsonian patients.
2. Correlative studies of C-11 methionine (MET) and
FDG PET in the evaluation of the tumor grading of
preoperative brain tumor and in the differentiation ofrecurrent brain tumor from radiation necrosis. For thedifferentiation between tumor recurrence and radiation necrosis, we are trying to decide cut-off value ofboth MET and FDG uptake, lesion to normal graymatter tissue count ratio (L/N).

Future Prospects
Imaging and Intervention (Diagnostic Radiology
and Nuclear Medicine)
1. Neruointerventional therapy for stroke and otherneurological disorders using Xray angio-MR systemwith monitoring diffusion/perfusion and vascular wallby MR.
2. Molecular imaging in the future
In the era of molecular medicine, molecular imaging is attracting a great deal of attention in the field ofimaging science. Molecular imaging approaches themolecular basis of biological processes using imaging
technology. We intend to perform researches inmolecular imaging. In particular, we are planning toinvestigate non-invasive imaging of gene expression
in living animals. We hope to contribute to developing
methods for gene expression imaging and, subsequently, advancing molecular biology and molecular
medicine.
3. Molecular Imaging in clinical oncology
The first point is the evaluation of angiogenesis
representing activity of primary neoplasms. Withadvent of CT- PET system, the degree of FDG uptake
will be correlated with the degree of angiogenesis.
Another point is the evaluation of lymph node metastasis. New contrast materials having affinity withmacrophages or tumor cells itself in the node will beavailable on MRI in the near future.
4. Functional imaging using CT and MRI
Functional imaging focusing on perfusion will be
useful especially in the liver and kidney among thebody. Another functional imaging is related to theactivity of hepatocytes and Kuppfer cells of the liverusing tissue-specific MR contrast agents.

5. Functional imaging using radiotracers
Measurements of endogenous neurotransmitter
secretion under various physiological conditions andpharmacological manipulations. We are planning tomeasure the amount of endogenous dopamine releasefrom the terminals of nigrostriatal dopaminergic neurons during executing mental tasks, drug therapy anddeep brain electrical stimulation in patients with
Parkinson’s disease, using C-11 raclopride and PET.
We are also planning to develop the new radiolabeledtracers for opiate receptors, dopamine, serotonine,
GABA and glutamate receptors and transporters forthe evaluation of mental functions in patients withpsychiatric disorders such as panic disorder, depression and schizophrenia.

Radiation Oncology
1.Image-guided real-time high dose-rate radiotherapy:
Continuous image acquisition during the therapeutic radiation visualizes errors in target localization andmotion of normal structures. In rotational conformal
therapy, a separate X-ray tube mounted on the linac
gantry enables real-time CT imaging of the target andsurrounding organs. When the system is combinedwith a newly investigated extremely high dose rate Xray source (laser X-ray source), time and spatial resolution of radiotherapy will be drastically improved. It
will greatly enhance radiosurgey application.
2.Tailor-made prediction of radiation sensitivity byuse of DNA array tip:
Genetic factors define the radiation sensitivity of
the normal tissue. Fibroblast cultures are performed
sporadically but take too long time in clinical practice.
DNA array analyses of single nucleotide polymorphism (SNP) may clarity how the sensitivity is determined and contribute to the decision of indicationand tailor-made dose setting for radiation therapy.

Research Grants (Department of Radiology)
The Grant for Development of Advanced Medical
Practice of The Ministry of Education, Science, Sports
and Culture in Japan.
1. Ohtomo K. Clinical necessity of both state-of-the art
CT and MR examinations in the evaluation of thesame regions of the body (2000-2002 ￥30.000.000)
The Grant-in-Aid for Scientific Research (B)(2) of The
Ministry of Education, Science, Sports and Culture in
Japan.
2. Abe O.: Cerebral perfusion MR imaging using continuous arterial spin labeling with 2-coil system. (20012004 ￥13.600.000)
3. Hayashi N. Optical coherence tomography of thesuperficial vascular diseases: fundamental study.
(2002-2005 ￥13.000.000)

Grant-in-Aid for Scientific Research (C) of The
Ministry of Education, Science, Sports and Culture in
Japan.
4. Nakagawa K. Megavoltage CT-assisted Stereotactic
Radiosurgery for Thoracic Tumors (1999-2001,¥3,500,000)
5. Nakagawa K. Development of computerized questionnaire system for collecting QOL information ofcancer patients (2000-2001, ¥3,400,000)

Select Publications (1997-2002, except for1.27.39) (Department of Radiology)
Diagnostic Radiology○1. Aoki S, Shirouzu I, Sasaki Y, Okubo T, Hayashi N,
Machida T, Hoshi E, Suzuki K, Funada N, Araki T,
Sasaki Y. Enhancement of the intracranial arterial
wall at MR imaging: Relation ship to cerebral atherosclerosis. Radiology 194:477-481,1995
2. Aoki S, Osawa S, Yoshioka N, Yamashita H, Kumagai
H, Araki T. Velocity-coded color MR angiography.
AJNR 19: 691-693,1998
3. Aoki S, Aoki K, Ohsawa S, Nakajima H, Araki T.
Dynamic MR images of the carotid wall. J Magnetic
Reson Imaging 9:420-427,1999
4. Aoki S, Nakajima H, Kumagai H, Araki T. Dynamic
contrast-enhanced MR angiography and MR imagingof the carotid artery: high-resolution sequences indifferent acquisition planes. AJNR 21:381-385,2000
5. Aoki S, Yoshikawa T, Hori M, Nanbu A, Kumagai H,
Nishiyama Y, Nukui H, Araki T. MR digital subtraction angiography for the assessment of cranial arteriovenous malformations and fistulas. AJR 175:451453,2000
6. Aoki S, Yoshikawa T, Hori M, Ishigame K, Nambu A,
Kumagai H, Araki T. 2D thick-slice MR digital subtraction angiography for assessment of cerebrovascular occlusive diseases. Eur Radiol 10:1858-1864,2000
7. Aoki S, Hayashi N, Abe O, Shirouzu I, Ishigame K,
Okubo T, Nakagawa K, Ohtomo K, Araki T.
Radiation-induced arteritis: thickened wall withprominent enhancement on cranial MR imagesreport of five cases and comparison with 18 cases ofmoyamoya disease. Radiology 2002 223:683-688
8. Hayashi N, Yamamoto S, Okubo T, Yoshioka N,
Shirouzu I, Abe O, et al. Avulsion injury of cervicalnerve roots: enhanced intradural nerve roots at MR
imaging. Radiology 1998;206(3):817-22.
9. Hayashi N, Tsutsumi Y, Barkovich AJ. Morphologicalfeatures and associated anomalies of schizencephaly
in the clinical population: detailed analysis of MR
images. Neuroradiology 44(5):418-27,2002.
○10. Hayashi N, Masumoto T, Abe O, Aoki S, Ohtomo
K, Tajiri Y. Accuracy of Abnormal Paraspinal Muscle
Findings on Contrast-enhanced MR Images as
Indirect Signs of Unilateral Cervical Root-Avulsion
Injury. Radiology 223(2):397-402,2002.

85

11. Hayashi N, Tsutsumi Y, Barkovich Polymicrogyria
without porencephaly/schizencephaly. MRI analysisof the spectrum and the prevalence of macroscopic
findings in the clinical population. Neuroradiology,
In print.
12. Yoshikawa T, Aoki S, Hori M, Nambu A, Kumagai H,
Araki T. Time-resolved two-dimensional thick-slice
magnetic resonance digital subtraction angiography
in assessing brain tumors. Eur Radiol 10: 736-744,
2000.
13. Yoshikawa T, Abe O, Tsuchiya K, Okubo T, Tobe K,
Masumoto T, Hayashi N, Mori H, Yamada H, Aoki S,
Ohtomo K. Diffusion-weighted magnetic resonanceimaging of the dural sinus thrombosis.
Neuroradiology 44: 481-488, 2002.
14.Abe O, Aoki S, Hayashi N, Yamada H, Kunimatsu A,
Mori H, Yoshikawa T, Okubo T, Ohtomo K. Normal
aging in the central nervous system:quantitative MR
diffusion-tensor analysis. Neurobiol Aging. 23:433441,2002.
○15. Abe O, Okubo T, Hayashi N, Saito N, Iriguchi N,
Shirouzu I, Kojima Y,Masumoto T, Ohtomo K, Sasaki
Y. Temporal changes of the apparent diffusion coefficients of water and metabolites in rats with hemispheric infarction: experimental study of transhemispheric diaschisis in the contralateral hemisphere at
7 tesla. J Cereb Blood Flow Metab. 20:726-735,2000
16. Mori H, Yoshioka H, Ahmadi T, Saida Y, Ohara K,
Itai Y. Early radiation effects on the liver demonstrated on superparamegnetic iron oxide-enhanced T1weighted MRI.
J Comput Assist Tomogr. 24(4):648-51,2000.
17. Mori H, Abe O, Okubo T, Hayashi N, Yoshikawa T,
Kunimatsu A, Yamada H, Aoki S, Ohtomo K.
Diffusion property in a hamartomatous lesion ofneurofibromatosis type 1. J Comput Assist Tomogr.
25(4):537-9,2001.
18. Okubo T; Yoshioka N; Hayashi N; Abe O; Masumoto
T; Sasaki T; Ohtomo K
Contrast-enhanced magnetic resonance imaging of
the cranial nerves in patients with acoustic schwannoma: correlation with surgical findings. Acta
Otolaryngol Suppl (Norway)542 :13-17,2000
19. Masutani Y et al. Automated Segmentation and
Visualization of the Pulmonary Vascular Tree in
Spiral CT Angiography: An Anatomy-Oriented
Approach based on Three-Dimensional Image
Analysis, J Comput Assist Tomogr 25(4):587-597,
2001
20. Masutani Y et al. Automated Segmentation ofColonic Walls for Computerized Detection of Polyps
in CT Colonography, J Comput Assist Tomogr
25(4):629-638, 2001
21. Matsuoka Y, Ohtomo K, Araki T, Kojima K,
Yoshikawa W, Fuwa S. MR imaging of clear cell carcinoma of the ovary. Eur Radiol 11: 946-951, 2001
22. Yoshioka N, Minami M, Inoue Y, Kawauchi N,
Nakajima J, Oka T, Yoshikawa K, Ohtomo K.
Pedunculated bronchogenic cyst mimicking pleurallesion. J Comput Assist Tomogr 24:581-583, 2000

86

23. Okada Y, Ohtomo K, Kiryu S, Sasaki Y. Breath-holdT2-weighted MRI of hepatic tumors: value of echo
planar imaging with diffusion-sensitizing gradient. J
Comput Assist Tomogr, 22:364-371, 1998.
24. Kobayashi Y, Minami M, Ohtomo K, Matsuoka Y.
MR imaging and CT appearance of aggressive
angiomyxoma. AJR 169:1752-1753,1997.
25. Minami M, Itai Y, Charnsangavej C, Ohtomo K,
Kawauchi N, Sasaki Y. “Periportal collar” sign of theliver: pathophysiologic approach. In: Selected scientific exhibits in gastrointestinal radiology: RSNA
1996 Radiographics 17(suppl 2): 1616-1620(abstr)
1997
26. Ohtomo K, Matsuoka Y, Abe O, Okada M, Amo K,
Minami M, Kawauchi N, Sasaki Y. High-resolution
MR imaging evaluation of hepatocellular carcinoma.
Abdom Imaging 22:182-186,1997.

Radiation Oncology○27. Nakagawa K, Aoki Y, Akanuma A, Onogi Y,
Terahara A, Sakata K, Muta N and Sasaki Y. Realtime beam monitoring in dynamic conformation
therapy. International Journal of Radiation Oncology,
Biology, Physics 30, 1233-1238, 1994
28. Nakagawa K, Aoki Y, Kusama T, Ban N and
Nakagawa S. Radiotherapy during pregnancy and its
effects on fetuses and neonates. Clinical
Therapeutics 19, 770-777, 1997
29. Nakagawa K, Aoki Y and Sasaki Y. Improvement ofimage quality in megavoltage computed tomography
with second generation scanning mode. Radiation
Oncology Investigation 5, 257-263, 1997
○30. Nakagawa K, Aoki Y, Fujimaki T, Tago M,
Terahara A, Karasawa K, Sakata K, Sasaki Y,
Matsutani M and Akanuma A. High dose conformal
radiotherapy influenced pattern of failure, but did
not improve survival of glioblastoma multiforme.
International Journal of Radiation Oncology, Biology,
Physics 40, 1141-1149, 1998
31. Nakagawa K, Aoki Y, Ohtomo K, Albright N andGoer D. Dosimetry of leakage doses from a mobile
accelerator for IORT and legal issues for its clinicaluse in Japan. International Journal of ClinicalOncology 4, 215-219, 1999
○32. Nakagawa K, Aoki Y, Tago M, Terahara A andOhtomo K. Megavoltage CT-assisted stereotactic
radiosurgery for thoracic tumors: original research in
the treatment of thoracic neoplasms. International
Journal of Radiation Oncology, Biology, Physics 48,
449-457, 2000
33. Tago M, Terahara A, Nakagawa K, Aoki Y, Ohtomo
K, Shin M and Kurita H. Immediate neurologicaldeterioration after gamma knife radiosurgery for
acoustic neurinoma. J Neurosurg 93, 78-81, 2000
34. Kozuka T, Aoki Y, Nakagawa K, Ohtomon K,
Yoshikawa H, Matsumoto K, Yoshiike K and Kanda
T. Enhancer-promoter activity of human papillomavirus type 16 long control regions isolated fromcell lines SiHa and CaSKi and cervical cancer biop-

sies. Japanese Journal of Cancer Research 91, 271279, 2000
35. Takeuchi T, Kozuka T, Nakagawa K, Aoki Y,
Ohtomo K, Yoshiike K and Kanda T. Adeno-associated virus type 2 nonstructural protein rep78 suppresses translation in vitro. Virology 266, 196-202, 2000
36. Nakagawa K, Kozuka T, Akahane M, Suzuki G,
Akashi M, Hosoi Y, Aoki Y and Ohtomo,K.
Radiological findings of accidental radiation injury of
the fingers. Health Physics 80, 67-70, 2001
37. Nakagawa K, Tago M, Terahara A, Aoki Y, Sasaki T,
Kurita H, Shin M, Kawamoto S, Kirino T and
Ohtomo K. A single institutional outcome analysis of
gamma knife radiosurgery for single or multiple
brain metastases. Clinical Neurology and
Neurosurgery 102, 227-232, 2001
38. Igaki H, Nakagawa K, Aoki Y, Ohtomo K, Kukimoto I
and Kanda T. Characterization of the Bi-directionaltranscriptional control region between the human
UFD1L and CDC45L genes. Biochemical and biophysical Research Communications 283, 569-576,
2001

ene diphosphonate after nephron sparing surgery. J
Nucl Med 40: 418-421, 1999
47. Inoue Y, Ohtake T, Homma Y, Yoshikawa K,
Nishikawa J, Sasaki Y Evaluation of glomerular filtration rate by a camera-based method in children andadults. J Nucl Med 39: 1784-1788, 1998
48. Inoue Y, Ohtake T, Yoshikawa K, Nishikawa J,
Sasaki Y Estimation of deadtime in imaging humansubjects.Eur J Nucl Med 25: 1232-1237, 1998
49. Inoue Y, Momose T, Ohtake T, Asai S, Yoshikawa K,
Nishikawa J, Sasaki Y Effect of deadtime loss on
quantitative measurement of cerebral blood flow
with technetium-99m hexamethylpropyleneamine
oxime. Eur J Nucl Med 24: 1418-1421, 1997
50. Inoue Y, Momose T, Ohtake T, Nishikawa J, Sasaki
Y, Waritani T, Inoue M Metabolism of technetium99m-L,L-ethyl cysteinate dimer in rat and cynomolgus monkey tissue.J Nucl Med 38: 1731-1737, 1997

Nuclear Medicine○39. Momose T, Sasaki Y, Sakurai Y, Iwata M.
Functional studies with O-15 H2O PET. Biomed
Research 13(Suppl 1); 77-82, 1992.
○40. Momose T, Nishikawa J, Watanabe T, Sasaki Y,
Senda M, Kubota K, Sato Y, Funakoshi M andMinakuti S. Effect of mastication on regional cerebral
blood flow in humans examined by positron emission tomography with O-15 labelled water and magnetic resonance imaging. Archs oral Biol. 42; 5761,1997
41. Oku S, Nakagawa K, Momose T, Kumakura Y, Abe
A, Watanabe T and Ohtomo K FDG-PET after radiotherapy is the best prognostic indicator of rectal cancer. Annals Nucl Med :16(6), 2002,in press.
○42. Inoue Y, Abe O, Kawakami T, Ozaki T, Inoue M,
Yokoyama I, Yoshikawa K, Ohtomo K Metabolism of99m
Tc-ECD in infarcted brain tissue of rats.J Nucl Med
42: 802–807, 2001
○43. Inoue Y, Yoshikawa K, Yoshioka N, Watanabe T,
Saegusa S, Kaneko Y, Yokoyama I, Ohtomo K
Evaluation of renal function with 99mTc-MAG3 using
semiautomated regions of interest.J Nucl Med 41:
1947-54, 2000
4
4. Inoue Y, Yoshikawa K, Suzuki T, Katayama N,
Yokoyama I, Kohsaka T, Tsukune Y, Ohtomo K
Attenuation correction in evaluating renal function
by a camera-based method in children and adults. J
Nucl Med 41: 823-829, 2000
4
5. Inoue Y, Ohtake T, Yokoyama I, Yoshikawa K, Asai
S, Ohtomo K Evaluation of renal function from technetium-99m-MAG3 renography without blood sampling.J Nucl Med 40: 793-798, 1999
46. Inoue Y, Ohtake T, Kameyama S, Yamazaki S,
Kawabe K, Yoshikawa K, Nishikawa J, Sasaki Y
Increased renal retention of technetium-99m methyl-87

Department of Radiation Oncology(Experimental Radiology)
Outline and Research Objectives
The department was founded in 1966 to carry out education and research on clarification of radiation effect on cells and tissues through biological and biochemical understanding.
Our recent research program(s) have focused on the basic studies for radiation oncology
and radiotherapy through understanding molecular biological basis of radiation responses incells and tissues.
The standardized radiotherapy has been daily fractionated irradiation of 2 Gy for about 6
weeks by which tumor cells lose clonogenic potential while normal tissues involved are
saved and recover. To clarify the mechanisms involved in the process and to devise bettermethods for radiotherapy have been the subject of vigorous study for decades. DNA has
been considered as the target for radiation action on the cells causing mitotic or reproductive cell death while a fertilized egg divides into various tissue cells and organs with thesame DNA but with different radiosensitivity. Tumors and normal tissues differ in radiosensitivity depending on intrinsic cellular radiosensitivity, cell kinetics, schedule of fractionation, type or quality of radiation used, radiation dose, assay systems used and physiologicalconditions such as pO2, pH, nutrient etc. However, the molecular mechanisms for the differences have been little clarified.
Toward this goal the specific projects aimed at the following problems have beenplanned and in progress.
Specific Aims:
1) To clarify biological mechanisms underlying radiation responses of normal tissues andtumors.
2) To clarify mechanism(s) and signal-transduction pathway(s) of cell death, tissue damage
and regeneration after irradiation and/or heat treatment, especially of apoptosis (programmed cell death)3) To establish predictive assay method(s) such as DNA and protein (antibody) arrays tohelp select treatment methods, i.e., type of radiation source(s) and irradiating protocol(s)suitable to individual patient and tumor.
4) To develop new methods for sensitization of tumor cells and for prevention of normal tissue damages.

Faculties and Students
Professor and Chair Norio Suzuki, M.D., Ph.D. (1986~)
Associate Professor Yoshio Hosoi, M.D., Ph.D.
Associates ...............................2
Graduate students...................2
Research students...................2

Past Research and Major Accomplishments
1. Quantitative assay system for mutation of cultured mammalian cells
2. Quantification of radiation induced mutation frequencies of mammalian cells in vitro or in vivo
3. Method to determine DNA synthesis point during
S-phase using mutation marker

88

4. Variability and instability of tumor cells in DNA
and various malignant properties
5. Clarification of metastatic processes and their relevance to radiation responses in mouse tumor system
6. Cell cycle dependence of metastatic lung colonization in mouse tumor system
7. New assay methods of metastasizing tumor cells
released into the blood of mice
8. Characterization of normal tissue stem cells especially testicular spermatogenic stem cells of mouse
9. Existence of hypoxic cells among mouse spermatogenic stem cells
10. Random lifetime and exponential decrease ofmouse spermatogenic stem cells

11. Activation of immune response by low dose irradiation
12. Inhibition of metastasis by low dose total body
irradiation
13. Sensitization of cells to ionizing radiation by
Chlorin e6Na

Current Research
Regarding radiation induced cell death, mitotic or
reproductive cell death has been traditionally considered significant and meaningful one underlying tumor
cure and tissue damages. DNA double-strand breaks(DSBs) are considered most harmful among various
DNA damages induced by ionizing radiation and theprimary cause for cell death. Thus, cellular repair
capacity of DSBs is a critical factor of radiosensitivity.
DSBs are mainly repaired by homologous recombination and non-homologous end joining (NHEJ). DNAdependent protein kinase (DNA-PK) is required for
NHEJ. We have previously reported that inactivationof DNA-PK by wortmannin or other drugs sensitizedcells to ionizing radiation.
On the other hand, tumor cells and normal tissue
cells have also been known to express apoptotic(interphase) cell death after irradiation and various
tumor cells are defective in apoptotic pathway andprocess. The role and significance of apoptotic cell
death in tumor cure or tissue damages depend on histological type or individual tumors and tissues.
We have been focusing currently on clarification of
signal transduction and process of cell death, especially apoptotic cell death after irradiation or heat treatment. T cell lymphoma, MOLT-4 cells (p53 wild), andmonoblastic U937 cells (p53 null) were mainly used.
The other focus has been clarification of the function
and the role of DNA-PK in signal transduction of DSB
repair and cell death.
1. Signal transduction mechanisms involved inradiation- or heat-induced cell death
As for involvement of receptor-mediated signaling
including Fas and TNFR, MOLT-4 cells stimulated bythese ligand did not undergo apoptosis, and irradiatedcells did not exhibit any mature form of caspase-8,
although Fas expression of MOLT-4 was upregulated
by irradiation. Dose dependency of p53 expression
and dominant negative effect of p53 on radiationinduced cell death of MOLT-4 cells have been reported. Expression of p53-related genes and the mechanism of the mitochondrial pathway are under-investigation.
1-1) Ceramide-JNK pathway
We have recently demonstrated the importantroles of ceramide-JNK pathway as well as p53 pathway in radiation-induced cell death of MOLT-4 cells;1) acid sphingomylinase inhibitor suppressed X-rayinduced apoptosis of MOLT-4 cells. 2) Rh-1a clone, aclone selected for radio-resistance from MOLT-4 cells,
was found resistant also to C2-ceramide-induced
apoptosis, with less activation of JNK. 3) the activation of JNK after X-irradiation or the treatment with a
JNK activator anisomycin caused the decrease of cMyc expression, 4) the reduction and/or inactivationof c-Myc by c-Myc inhibitor led to apoptotic cell
death.
We also demonstrated that the important roles ofceramide-JNK pathway in heat-induced cell death in
p53 null U937 cells by the transfection with the dominant negative c DNA of JNK into the cultured cells or
the treatment with acid sphingomylinase inhibitor.
We are studying further the mechanisms of heatinduced apoptosis and the role of ceramide-JNK signaltransduction pathway in the thermo-tolerance development.
1-2) p41-induction in radiation-induced apoptosis
p41, an acidic 41-kDa protein (pI=4.0) was newly
found in our laboratory as a radiation-induced protein
during apoptotic cell death of irradiated MOLT-4. The
protein was detected in two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and silver staining. The protein appeared radiation dose and timedependent. Amino acid sequence analysis of partial
peptides showed homology between p41 and a putative oncogene, set (also known as template activating
factor I, TAF-I). A polyclonal antibody was raised
against a synthetic partial peptide of p41.
Immunoblotting analysis of irradiated MOLT-4 cells
showed two spots, p41 and an additional 42-kDa protein, p42 (pI=4.1). p42 was detectable also in untreated cells. N-terminal amino acid sequencing of partiallypurified p41 and p42, and polyclonal antibodies
newly raised against different partial peptide
sequences revealed that p41 was a N-terminal truncation form of p42, and p42 was identified as SETb
(TAF-Ib), one of two SET isoforms. The cleavage site
was at carboxyl end of SNHD 18 of p42. A caspasespecific inhibitor or overexpressing of Bcl-2 suppressed radiation-induced p42 cleavage as well as
apoptotic cell death of MOLT-4. In vitro cleavage
experiments with recombinant p42 and either irradiated cell extracts or recombinant caspases, concluded
that the cleavage of p42 into p41 was catalyzed by
caspase(s) mainly by caspase-7.
One of newly raised antibodies specific to p41 or
specific to cleavage site of p42, was found usefulenabling simple detection of p41 by 1-D PAGE insteadof laborious 2-D PAGE. p41 would serve as a marker
of apoptotic cell death. The study on the role and sig-

89

nificance of p41-induction in radiation-induced apoptosis will be continued.
2. The function and the role of DNA-dependent
protein kinase (DNA-PK)
2-1) Target molecules and phosphorylation.
Although more than 30 proteins have been reported as DNA-PK substrates in vitro , it has been
unknown whether these were true targets in vivo. We
recently demonstrated XRCC4, a DNA ligase IV-associated protein, as the first example of in vivo substrate in response to radiation. We also identified a
new phosphorylation site in p53. Through the generation of phosphorylation-specific antibodies and theanalysis of phosphorylation-site-disruptants, we are
now attempting to clarify the role of the phosphorylation of XRCC4 and p53 in the repair and/or the signaltransduction of DNA double-strand breaks.
2-2) Hyperthermic lability and hyperthermic
radiosensitization
Radiosensitizing effect of hyperthermia has been
widely accepted and applied in cancer therapy.
Although many studies proposed various mechanisms
for the sensitization, the problem has not been solved
and still controversial. We examined hyperthermicstability of purified DNA-PK and its subunits. We
found and proposed heat lability of Ku subunit as apossible mechanism for hyperthermic radiosensitization. We also found that the heat stability of DNA-PK
was much higher in human cells than in rodent cells,
which may reflect the fact that many human cancer
cells are more refractory than mouse tumor cells tohyperthermia. We are now studying to modify stability of DNA-PK for improved hyperthermia-radiation
therapy.
2-3)Wortmannin affects apoptosis other than
repairHigher concentration of wortmannin such as 5μ
M or above, which were required for inhibition of
DNA-PK and ATM, caused enhanced radiation or heat
induced apoptotic cell death in MOLT-4 and V79 cells.
The apoptotic process overrode ongoing repair
process.
2-4) Prediction of radiation sensitivity.
We found that DNA-PK activity was high inhuman esophageal and colon cancer tissues comparedwith control normal tissues. The protein and mRNA
expressions of Ku70, Ku80, and DNA-PKcs were alsohigh in the esophageal and colon cancer tissues. The
protein and mRNA expressions of Ku70/80 correlatedwith DNA-PK activity. However, other studies including our own showed various results and the studies
are inconclusive.

902-5) Phosphorothioate oligonucleotide and its
analogues as inhibitiors of DNA-dependent protein kinase
We are currently studying the functions, the properties and the regulation mechanisms such as posttranslational modifications, of key enzymes in DSB
repair. We reported that phosphorothioate oligonucleotide and its analogue inhibited DNA-PK activity
and sensitized cells to ionizing radiation. The results
also showed 1) Correlation between DNA-PK activity
and radiation sensitivity 2)) Roles of DNA-PK and
ATM in cell-cycle-dependent radiation sensitivity 3)
Activation of epidermal growth factor receptor by ionizing radiation

Future Prospects
We hope that these studies will eventually lead us
to understanding mechanisms of difference in radiation responses among tissues and tumors, to the discovery and development of radiosensitizers/protectors, and also to the construction of gene/protein
array for predictive assay.

Research Grants
1. Scientific Research in Priority Areas (2) (1998-2000,
2001-2002)
P. I. Norio Suzuki"Molecular mechanisms of Radiation-responses andradiosensitivity"
2. Exploratory Research (2002-2003)
P. I. Norio Suzuki"DNA- and Protein-array for radiosensitivity"
3. Scientific Research in Priority Areas (2) (2001)
P. I. Yoshio Hosoi"Prediction of radiation sensitivity and radio-sensitization using DNA-dependent protein kinase"
4. Scientific Research in Priority Areas (2) (2001-2002)
P. I. Yoshihisa Matsumoto"The function of DNA-dependent protein kinase inradiation response and its possible application incancer radiation therapy I"
5. Grant-in-Aid for Young Scientists (B) (2001-2002)
P. I. Atsushi Enomoto"Analysis of the signal-transduction pathways andthe gene expressions using DNA micro-array systemon radiation effects"

Select Publications
1. Suzuki, N., Kawabe, S., Matsushima, T., and
Sugimura,T. Molecular species of aldolase in the liverof tumor-bearing rats. Gann 60, 433-437, 1969.

2. Sugimura, T., Sato, S., Kawabe, S., Suzuki, N., Chen,
T. C., and Takakura, K. Aldolase C in brain tumor.
Nature 222, 1070, 1969.
3. Suzuki, N., and Okada,S. Location of Ala 32 gene
replication in the cell cycle of cultured mammaliancells, L5178Y. Mutat. Res. 30, 111-116, 1975.
4. Suzuki, N., and Okada,S. Isolation of nutrient deficient mutants and quantitative mutation assay byreversion of alaniene-requiring L5178Y cells. Mutat.
Res. 34, 489-506, 1976.
5. Suzuki, N., and Okada,S. γ-Ray mutagenesis of cultured mammalian cells in vitro and in vivo.
Mutat.Res. 43, 81-90, 1977.
6. Suzuki, N., Withers, H. R., and Hunter, N.
Radiosensitization of mouse spermatogenic stem
cells by Ro-07-0582. Radiat.Res. 69, 598-601, 1977.
7. Suzuki, N., Frapart, M., Gardina, D. J., Meistrich, M.
L., and Withers, H. R. Cell cycle dependency ofmetastatic lung colony formation. Cancer Res. 37,
3690-3693,1977.
8. Nakamura, N., Suzuki, N., Harakawa, H., andOkada,S. Mutagenicity of furylfuramide, a food preservative tested by using alanine-requiring mouse
L5178Y cells in vitro and in vivo. Mutat.Res. 46,355364, 1977.
9. Okamura, S., Suzuki, N., Ishikawa, H., and Yamada,E. Quantitative changes in organelles during the cell
cycle of mouse leukemia cells as studied by electron
microscopy. Cell Struct. and Func. 2, 229-240, 1977.
10. Mian, T. A., Suzuki, N., Glenn, H. J., Haynie, T. P.,
and Meistrich, M. L. Radiation damage to mouse
testis cells from 99mTc pertechnetate. J. Nucl. Med.
18,1116-1122, 1977.
11. Suzuki, N., Withers, H. R., and Lee, L. Y. Variabilityof DNA content of murine fibrosarcoma cells. Nature
269, 531-532, 1977.
12. Suzuki, N., Withers, H. R., and Williams, M.
Radiation response in vitro in vivo of various clones
from a fibrosarcoma. Radiat.Res. 74, 503-504, 1978.
13. Suzuki, N., and Withers, H. R. Isolation from amurine fibrosarcoma of cell lines with enhanced plating efficiency in vitro. J.Natl.Cancer Inst. 60,179-183,
1978.
14. Suzuki, N., Withers, H. R., and Williams, M.
Heterogeneity and variability of artificial lung colony
forming ability among clones from a mouse offibrosarcoma. Cancer Res. 38, 3349-3351, 1978.
15. Meistrich, M, L., Hunter, N. R., Suzuki, N., Trostle, P.
K., and Withers, H.R. Gradual regeneration of mouse
testcular stem cells after ionizing radiation. Radiat.
Res. 74, 371-384, 1978.
16. Suzuki, N., and Withers, H. R. Exponential decraese
duringaging
andrandomlifetimeof mouse spermatogonial stem cells. Science 202,
1214-1215, 1978.
17. Suzuki, N., and Withers, H. R. Lung colony formation as a selective cloning process for lung colony
forming ability. Br. J. Cancer 39, 196-199, 1979.
18. Suzuki, N., Williams, M., Hunter, N. M., andWithers, H. R. Malignant properties and DNA con-tent of daughter clones from a mouse fibrosarcoma
differentiation among malignant properties. Br. J.
Cancer 42, 765-771, 1980.
19. Suzuki, N. Spontaneous versus artificial lung metastasis discrepant effect of whole-body irradiation in
NFSA2ALM and NFSA1SLM tumor systems. J. Natl.
Cancer Institute 71, 835-839, 1983.
20. Suzuki, N. New method to quantitate clonogenic
tumor cells in the blood circulation of mice. Cancer
Res. 43, 5451-5455, 1983.
21. Suzuki, N. Variant selection and blood-borne“Clonogenic” tumor cells in metastasis of FSA cellclones. Br. J. Cancer 48, 827-833, 1983.
22. Suzuki, N. Radiation response of “Clonogenic” tumorcell release from NFSA2ALM1 tumors. Radiat. Res.
98, 649-655, 1984.
23. Suzuki, N. Centrifugal elutriation and characterization of tumor cells from the venous blood of tumorbearing-mice possible relevance to metastasis. Cancer
Res. 44, 3505-3511, 1984.
24. Sakai, K., Suzuki, N., Nakamura, N., and Okada, S.
Induction and subsequent rapair of DNA damage by
fast neutrons in cultured mammalian cells.
Radiat.Res. 110, 311-320, 1987.
25. Suzuki, N., and Mizukoshi, T. Effect of low dosed ofwhole body irradiation on spontaneous lung metastasis of NFSA-2-ALM1 mouse tumors. Radiat.Med. 5,801-803, 1987.
26. Kobayashi, Y., Okabe, T., Urabe, A., Suzuki, N., andTakaku,F. Human granulocyte colony stimulating
factor produced by escherichia coli shortens the period of granulocytopenia induced by irradiation in
mice. Jpn. J. Cancer Res. 78, 763-766, 1987.
27. Sakata, K., Okada, S., Majima, H., and Suzuki, N.
Linear quadratic model of radiocurability on multicellular spheroids of human lung adenocarcinoma LCT1
and mouse ficrosarcoma FSA., Int. J. Radiat. Biol. 61,
269-274, 1992.
28. Majima, H., Kashiwado, K., Egawa, S., and Suzuki, N.
Interaction between the kinetics of thermotolerance
and effect of cis-diamminedichloroplatinum(II) or
bleomycin given at 37 ℃ or 43 ℃ . Int. J.
Hyperthermia 8, 431-442, 1992.
29. Kobayashi, S., Kasuya, M., Ishii, Y., Takehana, M.,
Sakai, K., Suzuki, N., and Itoi, M.
30. Effects of 2-mercaptopropionylglycine on the development of X-ray-induced cataract in rats. Current
Eye Reseach 11, 1099-1103, 1992.
31. Takakura, K., Ishikawa, M., Minegishi, A., Sakai, K.,
Suzuki,N., and Okada, S. Inverse dose-rate effect of
DNA breaks and inactivation of transforming activity induced by tritated water and 60Co gamma-rays.
Radiat. Envitron. Biophys. 31, 299-310,1992.
32. Hosoi, Y., and Kapp, Y. Expression of a candidate
ataxia-telangiectasia group D gene in cultured fibroblast cell lines and human tissues. Int. J. Radiat. Biol.
66, s71-s76, 1994.
33. Morimatsu, A., Suzuki, N., Hirano, K., Sakai, K., andMatsumoto, Y. Identification and characterization of91

X-ray-induced proteins in human T cell leukemia. J.
Radiat. Res. 37, 1-11, 1996.
34. Matsumoto, Y., Suzuki, N., Sakai, K., Morimatsu, A.,
Hirano, K., and Murofushi, H. A possible mechanism
for hyperthermic radiosensitization mediatedthrough hyperthermic lability of Ku subunits in
DNA-dependent protein kinase. Biochem. Biophys.
Res. Commun. 234, 568-572, 1997.
35. Hosoi, Y., Miyachi, H., Matsumoto, Y., Ikehata, H.,
Komura, J., Ishii, K., Zhao, H. J., Yoshida, M., Takai,Y., Yamada, S., Suzuki, N., and Ono, T. A phosphatidylionsitol 3-kinase inhibitor, wortmannin,
induces radioresistant DNA synthesis, and sensitizes
cells to bleomycin and ionizing radiation. Int. J.
Cancer 78, 642-647, 1998.
36. Matsumoto, Y., Umeda, N., Suzuki, N., Sakai, K., andHirano, K. A gel-electrophoretic analysis for
improved sensitivity and specificity of DNA-dependent protein kinase activity. J. Radiat. Res. 40, 183186, 1999.
37. Morita, A., Suzuki, N., Matsumoto, Y., Hirano, K.,
Enomoto, A., Zhu, J., and Sakai, K. p41 as a possible
marker for cell death is generated by caspase cleavage of p42/SETβ in irradiated MOLT-4 cells.
Biochem. and Biophys. Res. Commun. 278, 627-632,
2000.
38. Enomoto, A., Suzuki, N., Hirano, K., Matsumoto, Y.,
Morita, A., Sakai, K., and Koyama, H. Involvement of
SAPK/JNK pathway in X-ray-induced rapid cell deathof human T-cell leukemia cell line MOLT-4. Cancer
lett. 155, 137-144, 2000.
39. Matsumoto, Y., Suzuki, N., Namba, N., Umeda, N.,
Ma, X.-J., Morita, A., Tomita, M., Enomoto, A.,
Serizawa, S., Hirano, K., Sakai, K., Yasuda, and H.,
Hosoi, Y. Cleavage and phosphorylation of XRCC4
protein induced by X-irradiation. FEBS lett, 478, 6771, 2000.
40. Kang, Y., Hirano, K., Suzuki, N., Enomoto, A.,
Morita, A., Irimura, T., and Sakai,K. Increased
expression after X-irradiation of MUC1 in cultured
human colon carcinoma HT-29 cells. Jpn. J. Cancer
Res. 91, 324-330, 2000.
41. Tomita, M., Suzuki, N., Matsumoto, Y., Hirano,K.,
Umeda, N., and Sakai, K. Sensitization by wortmannin of heat-or X-ray induced cell death in cultured
chineses hamster V79 cells. J. Radiat. Res. 41, 93102, 2000.
42. Zhao, H.-J., Hosoi, Y., Miyachi, H., Ishii, K., Yoshida,M., Nemoto, K., Takai, Y., Yamada, S., Suzuki, N.,
and Ono, T. DNA-dependent protein kinase activity
correlates with Ku70 expression and radiation sensitivity in esophageal cancer cell lines. Clin. Cancer
Res. 6, 1073-1078, 2000.
43. Ono, T., Ikehata, H., Nakamura, S., Saito, Y., Hosoi,
Y., Takai, Y., Yamada, S., Onodera, J., andYamamoto, K. Age-associated increase of spontaneous mutant frequency and molecular nature ofmutation in newborn and old lacZ-transgenic mouse.
Mutat. Res. 447 165-177, 2000.

92

44. Nakamura, S., Ikehata, H., Komura, J., Hosoi, Y.,
Inoue, H., Gondo, Y., Yamamoto, K., Ichimasa, Y.,
and Ono, T. Radiation-induced mutation in spleen
and brain of plasmid-based lacZ transgenic mouse.
Int. J. Radiat. Biol. 76, 431-440, 2000.
45. Sakata, K., Matsumoto, Y., Tauchi, H., Satoh, M.,
Oouchi, A., Nagakura, H., Koito, K., Hosoi, Y.,
Suzuki, N., Komatsu, K., and Hareyama, M.
Expression of genes involved in repair of DNA double-strand breaks in normal and tumor tissues. Int. J.
Radiat. Oncol. Biol. Phys. 49, 161-167, 2001.
46. Sakata, K., Matsumoto, Y., Satoh, M., Oouchi, A.,
Nagakura, H., Koito, K., Hosoi, Y., Hareyama, M.,
and Suzuki, N. Clinical studies of immunohistochemical staining of DNA-dependent protein kinase inoropharyngeal and hypopharyngeal carcinomas.
Radiat. Med. 19, 93-97, 2001.
47. Enomoto, A., Suzuki, N., Liu, C., Kang, Y., Zhu, J.,
Serizawa, S., Matsumoto, Y., Morita, A., Ito, M., andHosoi,Y. Involvement of c-Jun NH2-terminal kinase1 in heat-induced apoptotic cell death of humanmonoblastic leukemia U937 cells. Int. J. Radiat. Biol.
77, 867-874, 2001.
48. Mori, N., Matsumoto, Y., Okumoto, M., Suzuki, N.,
and Yamate, J. Variations in Prkdc encoding the catalytic subunit of DNA-dependent protein kinase(DNA-PKcs) and susceptibility to radiation-induced
apoptosis and lymphomagenesis. Oncogene 20,
3609-3619, 2001.
49. Ishii, T., Futami, S., Nishida, M., Suzuki, T.,
Sakamoto, T., Suzuki, N., and Maekawa, K. Briefnote and evaluation of acute-radiation syndrome andtreatment of a tokai-mura criticality accident patient.
J. Radit. Res. 42, S167-S182, 2001.
50. Nakagawa, K., Kozuka, T., Akahane, M., Suzuki, G.,
Akashi, M., Hosoi, Y., Aoki, Y., Ohtomo, K.
Radiological findings of accidental radiation injury of
the fingers A case report. Healph phys. 80, 67-79,
2001.
51 Hosoi, Y., Matsumoto, Y., Tomita, M., Enomoto, A.,
Morita, A., Sakai, K.,Nakagawa, K., Umeda, N., Zhao,
H,-J., Ono, T., and Suzuki, N. Phosphorothioate
oligonucleotides, suramin and heparin inhibit DNAdependent protein kinase activity. Br. J. Cancer 86,
1143-1149, 2002.
52 Yoshida, M., Hosoi, Y., Miyachi, H., Ishii, N.,
Matusmoto, Y., Enomoto, A.,Nakagawa, K., Yamada,S., Suzuki, N., and Ono, T. Roles of DNA-dependent
protein kinase and ATM in cell-cycle dependent radiation sensitivity in human cells. Int. J. Radiat. Biol.
78, 503-512, 2002.
53 Komuro, Y., Watanabe, T., Hosoi, Y., Matsumoto, Y.,
Nakagawa, K., Kitayama, J., Suzuki, N., and Nagawa,H. The expression pattern of Ku correlates withtumor radiosensitivity and disease free survival inpatients with rectal carcinoma. Cancer 95, 11991205, 2002.

Department of
System Physiology
Outline and Research Objectives
This department was originally a part of the Research Institute of Medical Electronics
operated by the School of Medicine, but in 1997, when the University of Tokyo restructured
its system of education and research, the Institute became the Division of Biomedical
Engineering in the Graduate School of Medicine. The Division of Biomedical Engineering
consists of three departments: Bioimaging and Biomagnetics, Biosystem Construction andControl, and our own System Physiology.
This laboratory has been pursuing research on the biomechanics of phenomena in thehuman body, especially focusing on cellular sensing and response mechanisms to mechanical stimuli. The main theme of our work has been the relationships between the mechanicalforce generated by blood flow, shear stress and the cells exposed to it, vascular endothelial
cells. Research on this theme will be of benefit not only in understanding blood-flow-mediated regulation of vascular functions but in elucidating issues that are of importance clinically, such as angiogenesis, vascular remodeling, and atherogenesis, all of which occur in a
blood flow-dependent manner.
Original biomedical engineering methods have been applied in which cultured endothelial cells are exposed to controlled levels of fluid shear stress in a dynamically designed flow
apparatus (Fig. 1) and their responses are analyzed at the cellular and molecular levels.

Faculty and Students
Professor and Chair Joji Ando, M.D., Ph.D. (since 2000)
Lecturers
Masahiro Shibata, Ph.D.
Associates ...............................1
Graduate Students ..................3

Past Research and Major Accomplishments
Our studies have involved experiments on the following:
1. Endothelial cell responses to shear stress
2. Shear stress-mediated regulation of endothelial
gene expression
3. Shear stress signal transduction in endothelial cells
4. In vivo analysis of blood flow effects
And the results are described below.

Fig. 1.

Endothelial cell responses to shear stress
Our studies have demonstrated that endothelial
cells exhibit functional responses to shear stress.
When a cultured endothelial cell monolayer was partially denuded, surrounding cells migrated and proliferated in the denuded area, and covered it. Shearstress enhanced the regenerative functions ofendothelial cells (Microvasc Res 1987, Biorheology,1990), and it increased the production of nitric oxide,a potent vasodilator, in endothelial cells in a dosedependent manner (BBRC 1994). It also increased theexpression of thrombomodulin, an antithrombotic
molecule, in endothelial cells (BBRC 1994). By contrast, shear stress decreased the expression of vascular cell adhesion molecule-1(VCAM-1), which led toinhibition of leukocyte adhesion to endothelial cells(BBRC 1993, Am J Physiol 1994). A collaborative study

A silicon-tube-type flow-loading apparatus.

93manner (BBRC 1991, 1993). Flow-induced Ca 2+
responses generally start at a locus at the cell edge
and propagate throughout the entire cell in the formof a Ca 2+ wave (Fig. 3). The initiation locus corresponds precisely to caveola-rich cell edges (Proc Natl
Acad Sci 1998). We recently found that a subtype of
ATP-gated cation channel, the P2X4 receptor, isexpressed in human vascular endothelial cells andthat P2X4 receptors play a crucial role in the shear
stress-dependent Ca2+ response (Am J Physiol 2000,
Circ Res 2000).

showed that shear stress increases the levels of
adrenomedulin and C-type natriuretic peptide mRNA,
which have vasodilating effects (Hypertension 1997),
and that it augments the expression of low density
lipoprotein receptor (LOX-1) at both the protein andmRNA level (Circ Res 1998).
Shear-stress-mediated regulation of endothelial
gene expression
We have demonstrated that shear stress regulates
endothelial gene expression transcriptionally and/or
posttranscriptionally. It downregulates VCAM-1 genetranscription via the double AP-1 binding element(TGACTCA) in the promoter, which functions as ashear stress-responsive element (Am J Physiol 1997).
Fig.2 shows shear stress-induced changes in
VCAM-1 mRNA levels determined by the reverse
transcriptase PCR method. Shear stress has also beenshown
toincrease
the
level
of
granulocyte/macrophage-colony stimulating factor(GM-CSF) via mRNA stabilization (Circ Res 1988).
Differential display analysis showed that approximately 600 known and unknown transcripts are upor down-regulated in human umbilical vein endothelial cells exposed to a shear stress of 15 dynes/cm2 for
6 h (BBRC 1996), and a cDNA encoding an unknown
G-protein coupled receptor was cloned from theseshear stress-responsive genes (BBRC 1997).
Shear-stress signal transduction in endothelialcells
We were the first to show that Ca2+ signaling plays
an important role in the mechanism by which
endothelial cells recognize the shear stress signal andtransmit it into the cell interior (In Vitro Cell Dev Biol
1988) . Strong shearing forces induced by rubbing
endothelial cells with a balloon cause an increase incytoplasmic Ca2+ concentrations (Biorheology 1994). A
relatively weak shearing force, such as the shear
stress generated by fluid flow, requires the presenceof extracellular ATP to induce a Ca2+ response, and atseveral hundred nanomolar ATP, intracellular Ca2+
concentrations increase in a shear-stress-dependent94

Fig. 3. A flow-induced Ca2+ wave in an endothelial cell.

In vivo analysis of blood flow effects
Blood flow effects on capillary permeability tomacromolecules and angiogenesis have been investigated in vivo by intravital microscopy. We developed
a new intravital slit-laser microscanning method toquantitatively measure permeability to fluoresecentlabeled proteins at the distal end of arterioles, themiddle of capillaries and the proximal end of venules
in rabbit skeletal muscle tissue (Microvasc Res 1995,
1997). Permeability has been shown to increase withblood flow, indicating that shear stress regulates capillary protein permeability (Jpn J Physiol 1991, 1992).
We also investigated the relationship between capillary blood flow and tissue oxygen demand (Microvasc
Res 1985). We observed that increases in blood flow,which were induced by arterio-venous shunt oradministration of a-blocker, augmented the formationand development of new capillaries (Microvasc Res
1998). Our computer simulations have demonstrated
that formation of capillary networks in mammals corresponds well to an optimum model calculated fromthe oxygen supply efficiency in tissues (Microvasc Res
1995). An intravital laser microscope that utilizes aphosphorescence quenching technique was recently
developed to determine both microvascular and interstitial oxygen concentrations (J Appl Physiol 2001).

Current Research
Ongoing research in our laboratory include projects designed to 1) identify molecules that function asa flow sensor or shear-stress sensor in endothelial
cells, 2) analyze endothelial genes that respond toshear stress, 3) investigate effects of shear force on
the differentiation of endothelial progenitor cells, 4)
study the microcirculation.
Flow-sensing molecules: Our recent studies haverevealed that endothelial cells convert information on
shear stress into changes in intracellular Ca2+ concen-

trations, and that P2X4 purinoceptors and their ligand
ATP play an important role in the Ca2+ signaling. To
examine the role of P2X4 receptors as flow-sensors,
P2X4 cDNA was transfected into human embryonickidney (HEK) cells, and cell lines that stably express
P2X4 receptors were established. As shown in Fig. 4,
the control HEK cells did not show any Ca2+ response
to shear stress, whereas HEK cells that stably
expressed P2X4 receptors showed a shear stressdependent Ca2+ response. Thus, ectopic expression of
P2X4 receptors made the HEK cells sensitive to flow,
suggesting that P2X4 receptors function as a flow sensor. More recently, it has been demonstrated that
endothelial cells release ATP in response to shear
stress, and the endogenously released ATP is involvedin the activation of P2X4 receptors. Shear stress-sensing mechanisms are currently being investigated interms of ATP release.

recently observed that shear stress augments proliferation, differentiation, and tube-formation by EPCs.
This means that mechanical forces can manipulate thedifferentiation of EPCs. These techniques are ofpotential use in clinical applications, such as in thedevelopment of tissue-engineered vessels and celltherapy for ischemic vascular diseases.
Microcirculation study: Oxygen-sensing molecules that play an important role in the regulation ofvascular resistance are now being investigated, and an
in vivo optical method has been developed to monitor
pO2 distribution in skeletal muscle arterioles. We
have observed a significant drop in pO2 in the arterioles and a large pO2 gradient in the arteriolar walls,
indicating that the endothelial cells and/or smoothmuscle cells of arterioles may consume much more
oxygen than expected. We are also developing an insitu vascular Ca2+imaging system to explore the cellular mechanism of the effects of flow and hypoxia on
vascular tone.

Future Prospects
We intend to expand our research vascular biomechanics research to research on tissue engineering,atherosclerosis, and the vascular physiome in the near
future.

Fig. 4. P2X4 cDNA transfection makes HEK cells sensitive toshear stress.

Shear-stress-responsive genes: A high throughput
genomic analysis of gene expression using DNA
microarrays has been used to gain a more complete
appreciation of the extent and biologic significance ofendothelial activation by shear stress. Comparison of
the transcriptional activity of approximately 6,000
unique genes has revealed that laminar shear stressup- or down -regulates the expression of approximately 4% of the genes examined in endothelial cells.
Turbulent shear stress, which is closely related to theconditions under which atherosclerosis develops,
changed the expression of about 1.5% of the genes
examined. These genes included many of the genes
known to function in vascular remodeling, such as
genes encoding plasminogen activator, transforming
growth factor and metalloproteinase. Attention is
now focused on the role of these shear stress responsive genes in atherogenesis.
Effects of shear force on cell differentiation:
Endothelial progenitor cells (EPCs) have recently beenfound to circulate in blood vessels, migrate into tissues, and participate in neovascularization, and we

Vascular biomechanics: To elucidate the physiological or pathophysiological significance of shear
stress, shear stress-sensing molecules will be identified and knockout mice will be produced. Novel drugs
that can modulate the shear stress sensors will be
developed and applied to the treatment of vasculardiseases.
Tissue engineering: Techniques for manipulatingcell functions by mechanical forces will be developed
and applied to the preparation of tissue-engineeredblood vessels. A technique that allows reconstitutionof endothelial progenitor cells to a 3-dimensional, capillary-like structure under flow conditions will be used
to produce engineered capillary network tissue. The
capillary module will be expanded to an innovative
tissue engineering approach to produce blood-vesselcontaining engineered tissues, such as liver andmyocardial tissue.
Atherosclerosis research: Athero-prone or -protective genes will be identified among genes that
respond to turbulent shear stress. An in vitro modelof tissue-engineered atheroma will be established byarranging conditions, such as genetic background,
hemodynamics, and lipids related to atheroscelerosis.
The results of these studies may be useful in develop-95ing a novel therapy for atherosclerosis.
Vascular physiome: The term “physiome” isderived from “physio- (life)” and “-ome” (as a whole),
and means the quantitative and integrated descriptionof physiological functions based on information on
the genome, and proteome, and bioinformatics.
Mechanical forces generated in blood vessels regulate
vascular growth and remodeling in the human body.
An integrative description, including modeling, of themechanical force-mediated vascular controlling system will be conducted, and this is called the “vascularphysiome”.

Research Grants
1. Grants-in-Aid for Scientific Research from theJapanese Ministry of Education, Culture, Sports,
Science and Technology
Basic Research (A) 1995-1996 23,000, 000 Yen
2. Grants-in-Aid for Scientific Research from theJapanese Ministry of Education, Culture, Sports,
Science and Technology
Basic Research (A) 1997-1998 21,500, 000 Yen
3. Grants-in-Aid for Scientific Research from theJapanese Ministry of Education, Culture, Sports,
Science and Technology
Basic Research (A) 1999-2000 27, 900, 000 Yen
4. Special Coordination Funds for Promoting Scienceand Technology from the Japanese Ministry of
Education, Culture, Sports, Science and Technology
1998-2002 28, 000, 000 Yen
5. Grants-in-Aid for Scientific Research from theJapanese Ministry of Education, Culture, Sports,
Science and Technology
Basic Research (A) 2001-2002 28, 195, 000 Yen

Select Publications
1. M. Isshiki, J. Ando, K. Yamamoto, T. Fujita, Y. Ying,
R.G.W. Anderson: Sites of Ca2+ wave initiation movewith caveolae to the trailing edge of migrating cells.
J. Cell Sci. 115:475-484, 2002
2. R. Korenaga, K. Yamamoto, N. Ohura, T. Sokabe, A.
Kamiya, and J. Ando: Sp1-mediated downregulationof P2X4 receptor gene transcription in endothelial
cells exposed to shear stress. Am. J. Physiol Heart
Circ. Physiol. 280: H2214-H2221, 2001
3. M. Shibata, S. Ichioka, J. Ando, and A. Kamiya:
Microvascular and interstitial PO2 measurements in
rat skeletal muscle by phophorescence quenching. J.
Appl. Physiol. 91: 321-327, 2001
4. S. Kato, J. Ando, and T. Matsuda: mRNA expression
on shape-engineered endothelial cells: Adhesion
molecules ICAM-1 and VCAM-1. J. Biomed. Mater.
Res. 54:366-372, 2001
5. M. Negishi, D. Lu, Y-Q Zhang, Y. Sawada, T. Sasaki,
T. Kayo, J. Ando, T. Izumi, M. Kurabayashi, I.
Kojima, H. Masuda, and T. Takeuchi: Upregulatory

96

expression of furin and Transforming growth factorb by fluid shear stress in vascular endothelial cells.
Arterioscler Thromb Vasc Biol. 21:785-790, 2001
6. K. Yamamoto, R. Korenaga, A. Kamiya, and J. Ando:
Fluid shear stress activates Ca2+ influx into humanendothelial cells via P2X4 purinoceptors. Circ. Res.
87:385-391, 2000
7. K. Yamamoto, R. Korenaga, A. Kamiya, Z. Qi, M.
Sokabe, and J. Ando: P2X4 receptors mediate ATPinduced calcium influx in human vascular endothelial cells. Am. J. Physiol Heart Circ. Physiol.
279:H285-H292, 2000
8. J. Ando, R. Korenaga, and A. Kamiya: Shear StressDependent Regulation of Endothelial Cell Functions.
In “New Frontier in Vascular Biology; Thrombosis &Hemostasis”, K. Suzuki et al. eds., Eibun Press,
Osaka, pp199-215, 2000
9. J. Ando, R. Korenaga, and A. Kamiya: Flow-induced
endothelial gene regulation. In “Mechanical Forces
and the Endothelium”, P.I. Lelkes ed., Harwood
Academic Publishers, London, pp111-126, 1999
10. M. Nagase, J. Abe, K. Takahashi, J. Ando, S. Hirose,and T. Fujita: Genomic organization and regulationof expression of the lectin-like oxidized low-density
lipoprotein receptor (LOX-1) gene. J. Biol. Chem.
273:33702-33707, 1998
11. T. Murase, N. Kume, R. Korenaga, J. Ando, T.
Sawamura, T. Masaki, and T. Kita: Fluid shear stresstranscriptionally induces lectin-like oxidized LDL
receptor-1 in vascular endothelial cells. Circ. Res.
83:328-333, 1998
12. K. Masatugu, H. Itoh, T-H. Chun, Y. Ogawa, N.
Tamura, J. Yamashita, K. Doi, M. Inoue, Y.
Fukunaga, N. Sawada, T. Saito, R. Korenaga, J. Ando,and K. Nakao: Physiologic shear stress suppresses
endothelin-converting enzyme-1 expression in vascular endothelial cells. J. Cardiovasc. Pharmacol. 31:
S42-S45, 1998
13. M. Isshiki, J. Ando, R. Korenaga, H. Kogo, T.
Fujimoto, T. Fujita, and A. Kamiya: Endothelial Ca2+
waves preferentially originate at specific loci in caveolin-rich cell edges. Proc. Natl. Acad. Sci. U.S.A. 95:
5009-5014, 1998
14. H. Watanabe, R. Takahashi, X-X. Zhang, Y. Goto, H.
Hayashi, J. Ando, M. Isshiki, M. Seto, H. Hidaka, I.
Niki, and R. Ohno: An essential role of myosin lightchain kinase in the regulation of agonist- and fluidflow-stimulated Ca 2+ influx in endothelial cells.
FASEB J 12: 341-348, 1998
15. K. Kosaki, J. Ando, R. Korenaga, T. Kurokawa, and
A. Kamiya: Fluid shear stress increases the production of granulocyte-macrophage colony-stimulating
factor by endothelial cells via mRNA stabilization.
Circ. Res. 81:794-803, 1998
16. S. Ichioka, M. Shibata, K. Kosaki, Y. Sato, K. Harii,
and A. Kamiya: In vivo measurement of morphometric hemodynamic changes in the microcirculation
during angiogenesis under chronic a1-adrenergic
blocker treatment. Microvasc. Res. 55: 165-174, 1998

17. Y. Takada, C. Kato, S. Kondo, R. Korenaga, and J.
Ando: Cloning of cDNAs encoding G protein-coupled
receptor expressed in human endothelial cells
exposed to fluid shear stress. Biochem. Biophys. Res.
Commun. 240: 737-741, 1997
18. R. Korenaga, J. Ando, K. Kosaki, M. Isshiki, Y.
Takada, and A. Kamiya: Negative transcriptional regulation of the VCAM-1 gene by fluid shear stress inmurine endothelial cells. Am. J. Physiol. 273: C1506C1515, 1997
19. T. Chun, H. Itoh, Y. Ogawa, N. Tamura, K. Takaya,
T. Igaki, J. Yamashita, K. Doi, M. Inoue, K.
Masatsugu, R. Korenaga, J. Ando, and K. Nakao:
Shear stress augments expression of C-type natriuretic peptide and adrenomedulin. Hypertension
29:1296-1302, 1997
20. M. Miyasaka, H. Kawashima, R. Korenaga, and J.
Ando: Involvement of selectins in atherogenesis: A
primary or secondary event? Atherosclerosis IV:
Recent Advances in Atherosclerosis Research, NewYork Academy of Sciences 811:25-35, 1997
21. J. Ando, H. Tsuboi, R. Korenaga, K. Takahashi, K.
Kosaki, M. Isshiki, T. Tojo, Y. Takada, and A.
Kamiya: Differential display and cloning of shear
stress-responsive messenger RNAs in humanendothelial cells. Biochem. Biophys. Res. Commun.
225: 347-351, 1996
22. Y. Wang, W.S. Shin, H. Kawaguchi, M. Inukai, M.
Kato, A. Sakamoto, Y. Uehara, M. Miyamoto, N.
Shimamoto, R. Korenaga, J. Ando, and T. Toyo-oka:Contribution of sustained Ca2+ elevation for nitricoxide production in endothelial cells and subsequent
modulation of Ca2+ transient in vascular smooth muscle cells in coculture. J. Biol. Chem. 271:5647-5655,
1996
23. J. Ando and A. Kamiya: Flow-dependent regulationof gene expression in vascular endothelial cells. Jpn.
Heart J. 37: 19-32, 1996
24. A. Kamiya and J. Ando: Responses of vascularendothelial cells to fluid shear stress: Mechanism. In“Biomechanics-Functional Adaptation andRemodeling”, K. Hayashi et al. eds., Springer-Verlag,
Tokyo, pp29-56, 1996
25. H. Tsuboi, J. Ando, R. Korenaga, Y. Takada, and A.
Kamiya: Flow stimulates ICAM-1 expression timeand shear stress-dependently in human endothelial
cells. Biochem. Biophys. Res. Commun. 206:988-996,
1995
26. M. Shibata, T. Kawamura, M. Sohirad, and A.
Kamiya: A new fluorescence microscopy for tomographic observation of microcirculation by using dual
beam slit laser illumination. Microvasc. Res. 49: 300314, 1995
27. Y. Takada, F. Shinkai, S. Kondo, S. Yamamoto, H.
Tsuboi, R. Korenaga, and J. Ando: Fluid shear stressincreases the expression of thrombomodulin by cultured human endothelial cells. Biochem. Biophys.
Res. Commun. 205:1345-1352, 1994
28. W. Yang, J. Ando, R. Korenaga, T. Toyo-oka, and A.
Kamiya: Exogenous nitric oxide inhibits proliferationof cultured vascular endothelial cells. Biochem.
Biophys. Res. Commun. 203:1160-1167, 1994
29. J. Ando, H. Tsuboi, R. Korenaga, Y. Takada,
N.Toyama-Sorimachi, M. Miyasaka, and A. Kamiya:
Shear stress inhibits the adhesion of cultured mouseendothelial cells to lymphocytes by down-regulating
VCAM-1 expression. Am. J. Physiol. 267:C679-C687,
1994
30. J. Ando, A. Ohtsuka, Y. Katayama, R. Korenaga, C.
Ishikawa, and A. kamiya: Intracellular calcium
response to directly applied mechanical shearing
force in cultured vascular endothelial cells.
Biorheology 31:57-68, 1994
31. R. Korenaga, J. Ando, H. Tsuboi, W. Yang, I. Sakuma,
T. Toyo-oka, and A. Kamiya: Laminar flow stimulates ATP- and shear stress-dependent nitric oxide
production in cultured bovine endothelial cells.
Biochem. Biophys. Res. Commun. 198:213-219, 1994
32. A. Kamiya and J. Ando: Fluid shear stress and vascular endothelial cell biomechanics. In “Clinical
Biomechanics and Related Research”, Y. Hirasawa,
C.B. Sledge, S.L.-Y. Woo eds., Springer-Verlag Tokyo
pp. 255-271, 1994
33. A. Kamiya, R. Korenaga, and J. Ando: Endothelial cell
responses to fluid shear stress. In”EndotheliumDerived Factors and Vascular Functions”,T. Masakied., Elsevier Science Amsterdam pp. 103-112, 1994
34. J. Ando and A. Kamiya: Blood flow and vascularendothelial cell function. Frontiers Med. Biol. Engng
5:245-264, 1993
35. R. Korenaga, J. Ando, A. Ohtsuka, I. Sakuma, W.
Yang, T. Toyo-oka, and A. Kamiya: Close correlation
between cytoplasmic Ca++ levels and release of an
endothelium-derived relaxing factor from cultured
endothelial cells. Cell Struct. Funct. 18:95-104, 1993
36. A. Ohtsuka, J. Ando, R. Korenaga, A. Kamiya, N.
Toyama-Sorimachi, and M. Miyasaka: The effect offlow on the expression of vascular adhesion molecule-1 by cultured mouse endothelial cells. Biochem.
Biophys. Res. Commun. 193:301-310, 1993
37. J. Ando, A. Ohtsuka, R. Korenaga, T. Kawamura, and
A. Kamiya: Wall shear stress rather than shear rate
regulates cytoplasmic Ca++ responses to flow in vascular endothelial cells. Biochem. Biophys. Res.
Commun. 190:716-723, 1993
38. M. Shibata and A. Kamiya: Blood flow dependenceof local capillary permeability of Cr-EDTA in the rabbit skeletal muscle. Jpn. J. Physiol. 42: 631-639, 1992
39. M. Shibata, M. Yoshida and A. Kamiya:
Measurement of local capillary permeability in skeletal muscle by microscopic clearance method. Jpn. J.
Physiol. 41: 725-734, 1991
40. J. Ando, A. Ohtsuka, R. Korenaga, and A. Kamiya:
Effect of extracellular ATP level on flow-induced Ca++
response in cultured vascular endothelial cells.
Biochem. Biophys. Res. Commun. 179:1192-1199,
1991
41. O. Ono, J. Ando, A. Kamiya, Y. Kuboki, and H.
Yasuda: Flow effects on cultured vascular endothelial

97and smooth muscle cell functions. Cell Struct. Funct.
16:365-374, 1991
42. J. Ando, S. M. Albelda, and E. M. Levine: Culture ofhuman adult endothelial cells in liquid-liquid interfaces: A new approach to the study of cell matrix
interactions. In Vitro Cell Dev. Biol. 27A:525-532,
1991
43. J. Ando, A. Kawarada, M. Shibata, K. Yamakoshi,
and A. Kamiya: Pressure-volume relationships of finger arteries in healthy subjects and patients withcoronary atherosclerosis measured non-invasively byphotoelectric plethysmography. Jpn. Circ. J. 55:567575, 1991
44. J. Ando, S. Araya, Y. Katayama, A. Ohtsuka, A.
Kamiya: Flow-induced calcium response in cultured
vascular endothelial cells. In “The Regulation ofCoronary Blood Flow”, Inoue et al. eds., SpringerVerlag Tokyo pp 230-241, 1991
45. A. Kamiya, J. Ando, M. Shibata, and H. Wakayama:
The efficiency of the vascular -tissue system for oxygen transport in the skeletal muscle. Microvasc. Res.
39:169-185, 1990
46. J. Ando, T. Komatsuda, C. Ishikawa, and A. Kamiya:
Fluid shear stress enhances DNA synthesis in cultured endothelial cells during repair of mechanicaldenudation. Biorheology 27:675-684, 1990
47. J. Ando, T. Komatsuta, and A. Kamiya: Cytoplasmic
calcium response to fluid shear stress in cultured vascular endothelial cells. In Vitro Cell. Dev. Biol.
24:871-877, 1988
48. H. Nomura, C. Ishikawa, T. Komatsuda, J. Ando, and
A. Kamiya: A disk type apparatus for applying fluid
shear stress on cultured endothelial cell. Biorheology
25:461-471, 1988
49. A. Kamiya, J. Ando, M. Shibata, and H. Masuda:
Roles of fluid shear stress in physiological regulationof vascular structure and function. Biorheology
25:271-278, 1988
50. J. Ando, H. Nomura, and A. Kamiya: The effects of
fluid shear stress on the migration and proliferationof cultured endothelial cells. Microvasc. Res. 33:6270, 1987
51. M. Shibata, and A. Kamiya: Microcirculatory
responses to carotid sinus nerve stimilation at various ambient O2 tension in the rabbit tenuissimus
muscle. Microvasc. Res. 30: 333-345, 198598

Department of
Bioimaging and Biomagnetics
Outline and Research Objectives
Our department was established in 1974, with the objective of promoting research on
engineering science in medicine and biology. In April, 1994, Professor Shoogo Ueno joined
the University of Tokyo, replacing retiring Professor Masao Saito as chairman of theDivision of Engineering Science in the Institute of Medical Electronics. In April 1997, ourdivision re-established itself as the Department of Bioimaging and Biomagnetics. Since its
establishment in 1994, the Department of Bioimaging and Biomagnetics has been engaged
in research on engineering science in medicine and biology. Our department specializes inbiomagnetic research, which includes the measurement of biomagnetic fields, magnetic
stimulation of the brain, and the effects of magnetic fields on biological systems.

Faculties and Students
Professor and Chair Shoogo Ueno, Dr. Eng. (1994~)
Associate Professor Keiji Iramina, Dr. Eng.
Lecturer
Masakazu Iwasaka, Dr. Eng.
Postdoctoral Fellow ..............: 1
Graduate Student................: 15
Research Student ..................: 1
Secretary ...............................: 2

Past Research and Major Accomplishments
We have studied the measurement and control ofbiological systems based on biomagnetism. We develop, integrate, and apply new ideas through innovativeinterdisciplinary research approaches. Our research
topics cover wide areas in medical and biological engineering, including technology for measurement, imaging, and modelling in the following fields.
Magnetic stimulation of biological systems
Measurement of biomagnetic fields
Imaging of Electrical Information Based on
Magnetic Resonance Imaging (MRI)
Effects of magnetic and electromagnetic fields on
biological systems and materials(1) Transcranial magnetic stimulation (TMS) has
become an important tool for the study of the functional organization of the human brain. We developed
a method of localized and vectorial magnetic stimulation using a figure eight coil (Fig.1). This method facilitates stimulation of the motor cortex of the humanbrain within a 5 mm resolution. We studied the cortical excitatory and inhibitory systems, using the technique of paired TMS. We observed that TMS initiates
the excitation of both cortical interneurons andpyramidal tract neurons. Once the pyramidal tract
neurons generate a D-wave, the excitation is unaffected by a conditioning stimulus, which has an inhibitory effect on excitability of pyramidal tract neurons.
(2) For the measurement of biomagnetic fields, wedeveloped a SQUID (superconducting quantum interference device) system with high sensitivity that facilitates the measurement of auditory brainstem evokedmagnetic fields.
We studied higher brain functions associated withshort-term memory and mental rotation processes
using MEG. In the functional information related toshort-term memory processes, a DC-like slow wave
was observed in the period between latencies of 900

Fig.1 Basic principles of magnetic stimulation using a figure-eight coil. (a) A figure-eight coil on the head. (b) A single ring coil andan induced current pattern. (c) A figure-eight coil and an induced-current pattern.

99

msec and 1,500 msec during the short-term memorytask. A mental rotation process requires rotation andmatching of a pair of mental images. Dynamic properties of the electrical current distribution in the humanbrain that correspond to the early mental rotation
processes were investigated.
We also focused on the relationship between MEG
and fMRI and we studied the temporal and spatial
responses of both hemodynamic and neuronal activities. Activation of the primary somatosensory cortex
was investigated using MEG and fMRI. It was possible
with fMRI to discriminate between the area of thethumb and the ring finger in the primary somatosensory cortex. In MEG measurement, however, it is difficult to discriminate between two closely locateddipoles, if no initial information is given.
(3) Conventional MRI does not reveal information
about the electrical properties of the body. We developed new methods to visualize neuronal current distribution and electrical-impedance distribution. The
basic principle is to erase the effects of BOLD (blood
oxygenation level dependent) by subtracting MRI signals with different polarities of gradient magnetic
fields. Measurements were made with an echo planar
imaging (EPI) sequence at 1.5 T. MRI mapping of theneuronal currents in the brain during middle finger
and thumb tapping was clearly observed (Fig.2). A
new method for impedance tomography was introduced, based on MRI techniques. The basic idea ofimpedance imaging is to use the shielding effects of
induced eddy currents on spin precession.

(4) We investigated the dynamic behavior of waterin high gradient magnetic fields. A superconducting
magnet that produced 8 T magnetic fields at the center was used. The maximum product of the magnetic
field and the gradient was 400 T2 / m at z = 75 mm,
where the z-axis was directed along the bore axis. A
water chamber, 50 mm wide, 60 mm high, and 700
mm long was filled with distilled water. When thewater chamber was inserted into the bore, we
observed the phenomenon in which the water wasparted, and the bottom of the water chamber was
revealed. We call this phenomenon the Moses Effect.
A simple calculation shows that the magnetic force
acting on 100 ml of water at 20 ºC is 0.288 N or about
1/3 of the earth’s gravity, when exposed to a magnetic
field of 8 T and 50 T/m. Since the magnetic force acting on diamagnetic and paramagnetic materials moves
the materials along magnetic field gradients, any kindof biological cells and materials can be manipulated bymagnetic force. In contrast, when biological materials
such as fibrin and collagen are exposed to uniform
magnetic fields, parallel or perpendicular orientation
to the magnetic field direction is attained. For example, we successfully oriented adherent cells such assmooth muscle and endothelial cells, and osteoblasts
in parallel to the magnetic field direction after 8T
magnetic field exposure (Fig. 3). The magnetic manipulation and alignment of cells and other biologicalmaterials has opened a new horizon in tissue engineering.

(a) fMRI(b) current MRI
Fig.2 (a) fMRI and (b)MRI mapping of the neuronal currents in the brain during middle finger and thumb tapping.

Fig.3 Magnetically oriented (a) Fibrin (b) Collagen (c) Osteoblasts under an 8T static magnetic field.

100Current Research
Our department is conducting interdisciplinary
research in three major areas: imaging and cognitive
neuroscience, cell and tissue engineering, and theassessment of the biological effects of electromagnetic
fields.
Imaging and cognitive neuroscience
Our research focuses on the development of functional brain dynamics imaging with high time resolution and high spatial resolution. The main techniques
employed are TMS (transcranial magnetic stimulation), MEG (magnetoencephalography), EEG (electroencephalography), electric current imaging by MRI,
and impedance imaging by MRI. These techniques arenoninvasive and very useful for studying higher brain
functions of humans such as memory and cognition.
We developed a 2-channel repetitive transcranial
magnetic stimulator and an EEG measurement systemthat combines TMS. We were able to non-invasively
evaluate the cortical reactivity and functional connections between different brain areas. We used TMS toinvestigate memory encoding and retrieval, particularly the role of the dorsolateral prefrontal cortex inassociative memory for visual patterns. TMS disrupts
associative learning for abstract patterns over theright frontal area, which suggests that the participating cortical networks may be lateralized in accordancewith classic concepts of hemispheric specialization.
We compared current density distributions in electroconvulsive therapy (ECT) and TMS using the finite
element method. While the skull significantly affectedcurrent distributions in ECT, TMS efficiently induced
eddy currents in the brain. Our results will support
clinical investigations to determine the electrode andcoil positions that maximize efficacy.
To investigate the safety aspects of TMS on thebrain, functional and anatomical changes in the brain
were investigated. Our initial experimental results
revealed that TMS does not affect the fEPSP (field
excitatory post synaptic potential) of the rat hippocampus.
We introduced a new method of conductivity tensor imaging using diffusion-weighted MRI to obtain
conductivity tensor distribution of the brain.
Diffusion-weighted images were acquired withmotion probing gradients (MPGs) applied in three
directions. Conductivities in each MPG direction were
calculated from the fast component and the fractionof the fast component, and two-dimensional conductivity tensor was estimated. We also proposed a new
method of electrical current imaging based on the frequency shift technique of MRI.
In another study, we investigated a method for tissue characterization using diffusion tensor analysis

and applied external electrical currents.
Magnetophysiology
We studied the ischemic myocardial muscles of rat
using a high-resolution DC-SQUID magnetometer.
The information about the ischemic myocardium
region and changes of function were obtained bycaridiomagnetic imaging.
We also measured the magnetic fields associatedwith end plate potentials induced from neuromuscular junctions of frogs. Biomagnetic measurements ofsynaptic transmission processes by micro SQUID systems are useful for the physiological and pharmacological studies such as stimulus-excitation-contraction
coupling.
Biological effects of electromagnetic fields
We investigated the effects of strong magnetic
fields (8T, 14T) on the structure, organization, andfunction of biological systems and materials, and thepossible medical and therapeutic applications of magnetic fields.
The effects of 14 T strong static magnetic fields onthe functional properties of biological materials, suchas proteins, oxygen molecules, and water were also
investigated. (1) A spectrum profile of hemoglobin
inside red blood cells indicated conformationalchanges depending on the magnetic flux density. (2)Magnetic field exposure initiated platelet aggregation
with the aid of collagen and accelerated the aggregation of platelets and fibrin. (3) Leukemia cell proliferation was decreased by an 8 T magnetic field, as aresult of behavioural changes of dissolved oxygen during exposure. (4) Magnetic fields inhibited bioluminescence of luciferin and luciferease in both in vitro
and in vivo experiments. Possible applications of magnetic fields for the control of biochemical reactions
were suggested.
We explored the possible medical applications ofmagnetically oriented collagen. Magnetically aligned
collagen provides a scaffold for smooth muscle cells(A7r5), osteoblasts (MC3T3), nerve cells (PC12 andSchwann cells), and blood vessels on which to grow,and it directs the growth to a specific direction.
Further experiments, however, demonstrated themagnetic orientation of adherent cells without collagen guidance after long-term exposure to static magnetic fields. Further studies must be carried out toclarify the detailed mechanisms which include thediamagnetic properties of the cells. Our findings maylead to clinically viable treatments of bone fracturesand bone defects as well as medical engineering applications such as nerve regeneration.
With the ever-increasing worldwide use of mobile
phones in recent years, social concerns and anxieties
have been raised about the possible detrimental

101

effects on human health. We investigated the effects
of high frequency electromagnetic field exposure onrats.
We investigated the effects of exposure to thestandard frequency of electromagnetic waves used forcellular phones in Japan (1,439 MHz) on the permeability of the blood brain barrier (BBB), brain morphological changes, body mass fluctuations, and cognitive
functions and memory restoration of male SpragueDawley rats.
Our results suggest that exposure to a TDMA field
at levels much stronger than emitted by cellular
phones do not affect the learning and memory
processes when there are no thermal effects. In our
most recent study, however, we investigated theeffects of 1439 MHz TDMA electromagnetic fields on
sleep disturbance or melatonin synthesis in rats. The
pineal melatonin level decreased with short-term
TDMA exposure, whereas the serum melatonin level
was unchanged. Short-term TDMA exposure may
inhibit pineal melatonin synthesis in this limited
experiment. Further studies are currently underway
to confirm and explain our initial results.

erates or prevents damage to injured rat hippocampal
CA3 cells as well as our study on the magnetic orientation of Schwann cells for nerve regeneration.
Researches on the cellular responses to magnetic
fields will also proceed by analyzing the genetic
responses as well as the morphological changes ofcells under strong static magnetic fields. Magneticallyinduced cytoskeleton displacement in an intact cell
will be observed during both long and short-term
magnetic field exposures. A new technique for the
remote handling of macromolecules in living systems
by electromagnetic forces will also be developed.

Research Grants
1. 2000-2004 Grant-in Aid for Specially Promoted
Research, Ministry of Education, Science, Sports,
Culture and Technology, Japan (No. 12002002.)
386,000,000Yen “Study of Dynamic Brain Function
using Transcranial Magnetic Stimulation and
Neuronal Current Imaging”
2. 1998-2000 Grant-in Aid for Scientific Research (A),Ministry of Education, Science, Sports, Culture andTechnology, Japan (No. 10308037) 35,800,000Yen“Control of living systems by strong magnetic fields”

Future Prospects
In the next 3.5 years, we hope to achieve significant progress in our three major areas of research:
imaging and cognitive neuroscience, cell and tissue
engineering, and the assessment of biological effects
of electromagnetic fields.
By determining how the brain works, from thelevel of neurons to the relationships between complex
neural networks, a myriad of medical, therapeutic andengineering applications will be developed. The combination of neuromagnetic imaging techniques willallow us to understand the dynamic interactions
between individual cells and larger neural networks
that give rise to the patterns of electrical activity associated with higher brain function. We hope to determine where and how signals for various higher cognitive processes arise within the brain using TMS, EEG,
MEG, and MRI. The development of new imaging
methods to better visualize brain activity will also provide a wealth of new information. For example, our
new MR imaging techniques will be helpful for the
early detection of neurological disorders, such as acutecerebral infarctions. Ultimately, we hope the information retrieved will uncover the roots of neurologicaland psychiatric diseases.
By determining the effects of electromagnetic
fields on biological materials and systems and understanding the mechanisms involved, we can develop
many potentially viable cell and tissue engineering
applications. For example, we will continue ourresearch to determine whether repetitive TMS regen-102

Select Publications
1. Iramina K, Maeno T, Kowatari Y, and Ueno S: Effectsof transcranial magnetic stimulation on EEG activity.
IEEE Transactions on Magnetics, 38(5): in press,
2002.
2. Huang SF, Iramina K, Yumoto M, and Ueno S:
Visual-evoked magnetic fields associated with geometric and phonetic discrimination. IEEE
Transactions on Magnetics, 38(5): in press, 2002.
3. Kotani H, Kawaguchi H, Shimoaka T, Iwasaka M,
Ueno S, Ozawa H, Nakamura K, and Hoshi K: Strong
static magnetic field stimulates bone formation to adefinite orientation in vitro and in vivo. Journal of
Bone and Mineral Research, 17(10): 1814-1821, 2002.
4. Hong B, Kuwaki T, Kihwan J, Kumada M, Akai M,
and Ueno S: Changes in blood pressure and heart
rate by repetitive transcranial magnetic stimulationin rats. Neuroscience Letters, 329: 57-60, 2002.
5. Sekino M and Ueno S: Comparison of current distributions in electroconvulsive therapy and transcranial
magnetic stimulation. Journal of Applied Physics,
91(10), 8730-8732, 2002.
6. Epstein CM, Sekino M, Yamaguchi K, Kamiya S andUeno S: Asymmetries of prefrontal cortex in humanepisodic memory: Effects of transcranial magnetic
stimulation on learning abstract patterns.
Neuroscience Letters, 320(1-2): 5-8, 2002.
7. Uchida S, Goto K, Tachikawa A, Iramina K, and Ueno
S: Magnetocardiographic imaging for ischemic
myocardial muscles on rats. IEICE Transactions on
Information and Systems, 65-D(1): 30-35, 2002.

8. Niijima S and Ueno S: MEG source estimation using
the fourth order MUSIC method. IEICE Transactions
on Information and Systems, E85-D(1): 167-174,
2002.
9. Iwaki S, Tonoike M, and Ueno S: Visualization of thebrain activity during mental rotation processing
using MUSIC-weighted lead-field synthetic filtering.
IEICE Transactions on Information and Systems,
E85-D(1): 175-183, 2002.
10. Kasai K, Yamada H, Kamio S, Nakagome K, Iwanami
A, Fukuda M, Itoh K, Koshida I, Yumoto M, Iramina
K, Kato, M, and Ueno S: Brain lateralization for mismatch response to across- and within-categorychange of vowels. NeuroReport, 12(11): 2467-2471,
2001.
11. Iramina K, Kamei H, Yumoto M, and Ueno S: Effects
of repetition rate of electric stimulation on MEG and
fMRI signals. IEEE Transactions on Magnetics, 37(4):2918-2920, 2001.
12. Iwasaka M, Yaoita M, Kono T, and Ueno S:
Magnetically disturbed activity of immobilized catalase on an electrode in hydrogen-peroxide buffer.
IEEE Transactions on Magnetics, 37(4): 2941-2943,
2001.
13. Iwasaka M, Miyakoshi J, and Ueno S: Opticalabsorbance of hemoglobin and red blood cell suspensions under magnetic fields. IEEE Transactions on
Magnetics, 37(4): 2906-2908, 2001.
14. Iwasaka M, Miyakoshi J, and Ueno S:
Magnetophoresis of diamagnetic cells and microorganisms in a culture medium. IEEE Transactions on
Magnetics, 37(4): 2644-2646, 2001.
15. Ogiue-Ikeda M., Kotani H, Iwasaka M, Sato Y, andUeno S: Inhibition of leukemia cell growth undermagnetic fields of up to 8T. IEEE Transactions on
Magnetics, 37(4): 2912-2914, 2001.
16. Umeno A, Kotani H, Iwasaka M, and Ueno S:
Quantification of adherent cell orientation and morphology under strong magnetic fields. IEEE
Transactions on Magnetics, 37(4): 2909-2911, 2001.
17. Hayashi R, Imaizumi S, Mori K, Niimi S, Ueno S, andKiritani S: Elicitation of N400m in sentence comprehension due to lexical prosody incongruity.
NeuroReport, 12(8): 1753-1756, 2001.
18. Iwasaka M, Takeuchi M, and Ueno S: Aggregation ofblood platelets in static magnetic fields. IEEE
Transactions on Magnetics, 36(5): 3721-3723, 2000.
19. Uchida S, Iramina K, Goto, K, and Ueno S: A comparison of iterative minimum norm estimation and current dipole estimation for magnetic field measurements from small animals. IEEE Transactions on
Magnetics, 36(5): 3724 -3726, 2000.
20. Tsuda R and Ueno S: Source localization of visually
evoked magnetic fields to stimuli in apparent
motion. IEEE Transactions on Magnetics, 36(5):3727-3729, 2000.
21. Tsurita G, Nagawa H, Ueno S, Watanabe S, and Taki
M: Biological and morphological effects on the brain
after exposure of rats to a 1439 MHz TDMA field.
Bioelectromagnetics, 21: 364-371, 2000.

22. Ichioka S, Minegishi M, Iwasaka M, Shibata M,
Nakatsuka T, Harii K, Kamiya A, and Ueno S: Highintensity static magnetic fields modulate skin microcirculationandtemperatureinvivo .
Bioelectromagnetics, 21: 183-188, 2000.
23. Liu R and Ueno S: Calculating the activating functionof nerve excitation in inhomogeneous volume conductor during magnetic stimulation using the finite
element method. IEEE Transactions on Magnetics,
36(4): 1796-1799, 2000.
24. Kotani H, Iwasaka M, Ueno S, and Curtis A:
Magnetic orientation of collagen and bone mixture.
Journal of Applied Physics, 87(9): 6191-6193, 2000.
25. Uchida S, Iramina K, Goto K, and Ueno S: Current
source imaging for high spatial resolution magnetocardiography in normal and abnormal rat cardiac
muscles. Journal of Applied Physics, 87(9): 62056207, 2000.
26. Tsurita G, Ueno S, Tsuno N, Nagawa H, and Muto T:
Effects of exposure to repetitive pulsed magnetic
stimulation on cell proliferation and expression ofheat shock protein 70 in normal and malignant cells.
Biochemical
andBiophysicalResearch
Communications, 261(3): 689-694, 1999.
27. Iramina K, Kamei H, Uchida S, Kato T, Ugurbil K,
Yumoto M, and Ueno S: Effects of stimulus intensity
on fMRI and MEG somatosensory cortex using electrical stimulation. IEEE Transactions on Magnetics,
35(5): 4106-4108, 1999.
28. Kamei H, Iramina K, Yoshikawa K, and Ueno S:
Neuronal current distribution imaging using magnetic resonance. IEEE Transactions on Magnetics, 35(5):4109-4111, 1999.
29. Yukawa Y, Iriguchi N, and Ueno S: Impedance magnetic resonance imaging with external AC field
added to main static field. IEEE Transactions on
Magnetics, 35(5): 4121-4123, 1999.
30. Uchida S, Iramina K, Goto K, and Ueno S: High-resolution magnetocardiography for the study of dynamic propagation of excitation sites in rat cardiac muscles. IEEE Transactions on Magnetics, 35(5): 41244126, 1999.
31. Nakagawa S and Ueno S: Measurements and source
estimations of extremely low frequency brain magnetic fields in a short-term memory task by a wholehead neurogradiometer. IEEE Transactions on
Magnetics, 35(5): 4130-4132, 1999.
32. Nakagawa S, Ueno S, and Imada T: Changes inparameters pf auditory evoked magnetic fields withclick train patterns. Journal of Applied Physics, 85(8):5708-5710, 1999.
33. Sugase Y, Yamane S, Ueno S, and Kawano K: Global
and fine information coded by single neurons in thetemporal visual cortex. Nature, 400(6747): 869-873,
1999.
34. Iwaki S, Ueno S, Imada T, and Tonoike M: Dynamic
cortical activation in mental image processing
revealed by biomagnetic research. NeuroReport, 10:
1793-1797, 1999.

10335. Ueno S: Biomagnetic approaches to studying thebrain. IEEE Engineering in Medicine and Biology, 18:
108-120, 1999.
36. Ueno S and Iriguchi N: Impedance magnetic resonance imaging: a method for imaging of impedance
distributions based on magnetic resonance imaging.
Journal of Applied Physics, 83(11): 6450-6452, 1998.
37. Iramina K, Hong B, Uchida S, Goto K, and Ueno S:
Measurements of biomagnetic fields using a high-resolution dc superconducting quantum interference
device magnetometer. Journal of Applied Physics,
83(11): 6465-6467, 1998.
38. Iwaki S and Ueno S: Weighted minimum-norm
source estimation of magnetoencephalography utilizing the temporal information of the measured data.
Journal of Applied Physics, 83(11): 6441-6443, 1998.
39. Iwasaka M and Ueno S: Structure of water molecules
under 14T magnetic field. Journal of Applied Physics,
83(11): 6459-6461, 1998.
40. Mihara H, Iriguchi N, and Ueno S: A method of rf
inhomogeneity correction in MR imaging. MR
Materials in Physics, Biology and Medicine, 7: 115120, 1998.
41. Iwasaka M, Takeuchi M, Ueno S, and Tsuda H:
Polymerization and dissolution of fibrin underhomogenous magnetic field at 14T. Journal ofApplied Physics, 83(11): 6453-6455, 1998.
42. Iwasaka M and Ueno S: Bioluminescence under static
magnetic fields. Journal of Applied Physics, 83(11):6465-6458, 1998.
43. Ichioka S, Iwasaka M, Shibata M, Harii A, Kamiya A,
and Ueno S: Biological effects of static magnetic
fields on the microcirculatory blood flow in vivo: apreliminary report. Medical and BiologicalEngineering & Computing, 36: 91-95, 1998.
44. Kobayashi M, Hyodo A, Kurokawa T, and Ueno S:
The first component of polyphasic motor evoked
potentials is resistant to suppression by paired transcranial magnetic stimulation in humans. Journal of
the Neurological Sciences, 159: 166-169, 1998.
45. Iramina K, Yumoto M, Kamei H, and Ueno S:
Measurement of somatosensory evoked responseusing functional MR images and MEG. IEEE
Transactions on Magnetics, 33(5): 4260-4262, 1997.
46. Iwaki S and Ueno S: A high-speed estimation ofinternal electrical sources in the human brain from
the MEG measurement using subspace scanningwith multiple scanning resolutions. IEEE
Transactions on Magnetics, 33(5): 4272-4274, 1997.
47. Kobayashi M, Ueno S, and Kurokawa T: Importanceof soft tissue inhomogeneity in magnetic peripheral
nerve
stimulation
mechanism.
Electroencephalography
andClinical
Neurophysiology, 105: 406-413, 1997.
48. Iramina K and Ueno S: Source estimation of spontaneous MEG activity and auditory evoked responses
in normal subjects during sleep. Brain Topography,
8(3): 297-301, 1996.
49. Fujiki M, Isono M, Hori S, and Ueno S: Corticospinal
direct response to transcranial magnetic stimulation

104

in humans. Electroencephalography and Clinical
Neurophysiology, 101: 48-57, 1996.
50. Iwasaka M, Ueno S, and Tsuda H: Measurement ofclottability of fibrin polymers using magnetic orientation. Journal of Applied Physics, 79(8): 4708-4710,
1996.

Department of Biosystem
Construction & Control
Outline and Research Objective
This department was established in 1964 as Division of Clinical Medicine in Institute ofMedical Electronics for research and development of the advanced diagnostic and therapeutic instruments for the clinical medicine and their related technologies. The education to
train the leaders of biomedical engineer and clinical engineer is its another important role.
The first The name of the department and institute has been changed as shown above sinceApril 1, 1997 with the structural reform of Faculty of Medicine.
This department has been cooperating with Division of Artificial Biomechanism,Research Center for Advanced Science and Technology (RCAST) presided by Associate Prof.
Chinzei and Division of advanced Biomedical Engineering & Life Science, Center for
Cooperative Research (CCR) presided by Prof. Mabuchi in the recent 10 years. Both laboratories, 10 Km distant, are connected with a personal microwave television system for all-day
long to exchange many information, to have a periodical research meeting and to monitor
the artificial heart animals from each other.
Our research fields are artificial heart, artificial valve, biomaterials, medical laser, medical
thermography, measurement instruments and sensors, and micromachine.

Faculties and Students
Professor and Chair Kou Imachi, Ph. D.(1993 -)Associate Professors Yusuke Abe, M. D., Ph. D.
Lecturer
Takashi Isoyama, Ph. D.
Associate .................................1
Technical Assistant ................1
Graduate Students ..................2
Guest Researchers...................6
Secretary .................................1
Professor Emeritus Kazuhiko Atsumi, M. D., Ph. D.,
Iwao Fujimasa, M. D., Ph. D.

Past Research and Major Accomplishments
Professor Imachi joined to Institute of Medical
Electronics in 1970 as a student of Graduate School of
Medicine, University of Tokyo, after he got the master’s degree of mechanical engineering at Kyoto
University and spent 2 years at Hitachi Chemical Co.
Ltd. as a chemical plant research engineer. Since thattime he has been studying biomedical engineeringand he became Associate in 1972, Associate professor
in 1987 and Professor in 1993. The past research andmajor accomplishments performed by him and his
group are shown in the following;
1. Artificial heart
The artificial heart(AH) study is world famous
research project having a long history since 1959.
Almost all the research fields such as driving mechanism, blood pump, artificial valve, materials, measure-ment & control method, circulatory physiology andpathophysiology have been developed and studied.
They are summarized in Figure1.
Our peculiar policy for AH research is shown in
Table 1. According to the policy, we have been
approaching to an implantable total AH(TAH) withquite different way from other groups and haveobtained many accomplishments summarized inTable 2. Among these accomplishments, trying 4
types of TAH, development of jellyfish valve, deviceof 1 / R control method, 3 times of the world longest
survival record under TAH goat, proposal of many
pathophysiological hypothesis, and invention ofundulation pump are the most unique and remarkable
outputs.
2. Biomaterials
Concerning the biomaterials, the mechanism ofthrombus formation and calcification on the medical
polymer surfaces, especially in the artificial heart
blood pump and on polymer valve have been studied.
The evaluation of biocompatibility of many biomaterials was also important task.
3. Medical laser
Medical laser research in this laboratory was started in 1965. Ruby laser, CO 2 laser, Nd-YAG laser,
excimer laser, argon laser, semiconductor laser havebeen studied. Laser scalpel, laser coagulator, laser
endoscopes for percutaneous intradiscal laser
nucleotomy, for colonary laser angioplasty and for
percutaneous mitral valvotomy, were developed. As

105

Table 1 The policy of AH research in our laboratory
Never copy the foreign countries project (respect the other person’s originality)
Don’t be captured by the existing laws and theories(there is no common sense in interdisciplinary field research)
Don’t cling to your background
Have always hypothesis
Don’t change the system until it runs into a stone wall
Don’t change more than 2 parameters at once
Try to make all the things by yourself (Speed is the most important)
Don’t hesitate to publish the negative data (truth is hidden in the negative data)

Table2 Accomplishments in AH research in this laboratory
Four types of TAH,: FTAH(1963-1980), HTAH(1977-1982), TRAH(1981-1996) and ITAH(1996-)
FTAH: TAH with fibrillated natural heart
HTAH: Hybrid TAH, Natural heart is beating but trunks of PA and Ao is occluded
TRAH: Total replacement AH, NH is resected and 2 pumps are placed on the chest wall
ITAH: Implantable TAH, All the components except energy source are implantedHome made artificial valves
OED(Oblique elliptical seat door type) valve 1971 - 1978
Jellyfish valve: polymer membrane valve(1987 - )Control method
Fixed cardiac output
Fixed cardiac output with electrical stimulation
1 / R control
World longest survival record with TAH in goat
1980: 288 days with HTAH
1984: 344 days with TRAH
1995: 532 days with TRAH under 1 / R control
Hypothesis
Hyper cardiac output syndrome: 1973 - 1975
Cardiac receptor hypothesis:: 1982 - 1895
Calcification mechanism hypothesis: 1995 - 2000
Blood pump & materials
Critical surface roughness to promote thrombus for mation(3 mm): 1970
Surface coating of segmented polyurethane on PVC: 1973 Multi-material test method of blood compatibility: 1984 - 1990
Clinical device development
Pneumatic driven ventricular assist device in cooperation with Aisin Seiki & Nippon Zeon Co. Ltd.:1990 -106for the basic research, the influence of various kindsof lasers on living tissue, laser therapy for tumor andcancer, laser neurotomy, bactericidal effect of laser
were investigated.
4. Medical thermography
Research of medical thermography in this laboratory was started in 1965. Since that time, many kindsof infrared thermography for clinical use has beendeveloped in cooperation with Fujitsu, Nihon-denshi,NEC, Nihonkoden Co.Ltd, etc.. As the basic and application researches, theoretical model construction toestimate subcutaneous blood flow from thermogram,digital processing of thermogram, dynamic thermography, standardization of clinical daignosis, thermotome, thermo-myotome, etc were studied.
5. Measurement instruments and sensors
Portable electro-magnetic flow(EMF) meter, EMF
flow probe for AH, blood lactate measurement instrument, remote monitoring system of AH animals
through microwave TV, implantable CCD probe toobserve microcirculation were developed.
6. Micromachine
Micromachine research was started in 1987. The
word “micromachine” was made in this laboratorywhen we began to start a society for the study of
micro-mechanical system in 1988. Artificial muscle
moved by vibrating energy, regenerative nerve electrode, micro-laser endoscope were studied and developed

Current Research
1. Artificial heart
Since 1996, AH project in this laboratory is
focused on the development of implantable TAH in
which the all components except outer battery andenergy transmission system are implanted into thebody and expected as the substitution of the hearttransplantation. Undulation pump TAH(UPTAH)
which is the most compact TAH in the world wasinvented(Fig. 2). Another merit of UPTAH is that it
can produce any kinds of flow patterns such as
pusatile flow, continuous flow and pulsatile flow on
the continuous flow, etc.. At the present stage, 6 goatssurvived over 1 month including 63 days as thelongest. Now all the components such as blood pump,driving unit, control system, measurement systemand energy transmission system are being made morecompact and increased in durability and reliability.
The influences of nonpulsatile flow on microcirculation and organ function are being in research. A new

Fig.2 Assembled UPTAH

project to develop an implantable ventricular assist
device for clinical use for the bridge to heart transplantation has been ready to start using the undulation pump. According to our calcifiction hypothesis,
the jellyfish valve is improved design to reduce thecreep fatigue. The new designed valve proved almost
10 years durability in-vitro acceleration fatigue testor.
2. Other researches
In the laser research, handy type laser blood coagulator aiming to use in the surgical field, is being
developed and evaluated in the animal experiments.
The imaging of arterial blood flow distribution in theskin and organs using a high speed thermography is
studying. CCD probe to observe microcirculation
chronically is being made compact including focus.
Regenerative nerve electrode to control artificialorgans through nervous signals is improving its tissuecompatibility.
3. New research
Tissue engineering research has started in thisyear. As the first step, valve membrane of jellyfish
valve is trying to make using endothelial cell culture.

Future Prospects
The following researches are scheduled as the
future projects and a part of them is already ready tostart.
Development of a permanent use artificial heart tobe able to substitute the heart transplantation. A
rotary type undulation pump which moves magnetically suspended is begun to design.
Complete the regenerative nerve electrode andfind a control algorithm of artificial organs throughnervous signals.
Development distributed artificial heart system inwhich the artificial blood pumps are connected toeach organs and tissues. Basic study to connect asmall AH to a kidney has already started.

107Regenerative medicine to make a whole organs
from ES cells will be started aiming to how to shortening the growing time utilizing a biomedical engineering technologies.

Research Grants in these 5 Years
1. The Program for Promotion of Fundamental Studies
in Health Science of the Organization for
Pharmaceutical Safety and Research(OPSR) “ Thecomprehensive basic research on the development ofJapanese original implantable total artificial heart”,
Head Investigator: K. Imachi, 170,460,000yen, 1996
– 20002. Grant-in-Aid for Scientific Research (B)(2):” A study
on long-term and continuous observation and analysis of microcirculation under artificial circulation”,
Top Head Investigator: K. Imachi, 16,000,000yen,
1999 – 2000
3. Grant-in-Aid for Scientific Research (A)(2): “Development of a simplified pulsatile left ventricularassist device”, Head Investigator: K. Imachi,
17,200,000yen, 1999 – 2000
4. Grant-in-Aid for Scientific Research (A)(2):”Researchand development of implantable artificial heart forpermanent use”, Head Investigator: Y. Abe,
42,600,000yen, 2000 – 2002
5. Grant-in-Aid for Scientific Research (S):“Comprehensive research for the influence of flow
state of artificial heart on the living body by observation of microcirculation”, Head Investigator: K.
Imachi, 87,900,000yen, 2002 – 2006

Select Publications
1. K. Atsumi, Y. Sakurai, I. Fujimasa, K. Imachi, T.
Nishisaka, I. Mano, H. Ohmichi, J. Mori, N. Iwai and
A. Kouno: Hemodynamic analysis on prolonged survival cases (30 days and 20 days) of artificial total
heart replacement, Trans. Am. Soc. Artif. Intern.
Organs, 21,545-554, 1975.
2. I. Mano, I. Fujimasa, K. Imachi, T. Nishisaka, H.
Ohmichi, J. Mori, N. Iwai, K. Atsumi, Y. Sakurai andI. Kino: Pathophysiological studies of the animals
replaced with the artificial heart. Part II. Concerning
liver,, gastrointestinal tract, kidney, and general discussion, Japanese Heart Journal, 17(3),326-337, 1976.
3. K. Imachi, I. Fujimasa. T. Nishisaka, H. Ohmichi, N.
Iwai, A. Koun, H. Miyake, and K. Atsumi:How toprevent thrombus formation in the artificial heart,
Artificial Organs, 2(Suppl)
4. I. Fujimasa, K. Imachi, H. Ohmichi, S. Takasugi, T.
Nsisaka, N. Iwai, A. Kuno, H. Miyake and K. Atsumi:
A circulatory dynamic model for artificial heart control, Artificial Organs, 2(Suppl.), 214-216, 1978.
5. H. Ohmichi, K. Imachi, I. Fujimasa, T. Nisisaka, N.
Iwai, A. Kouno and K. Atsumi: Hemodynamic andblood gas analysis in total artificial heart replaced
animal during treadmill exercise, Artificial Organs,2(Suppl.) 230-233, 1978.

108

6. K. Atsumi, I. Fujimasa, K. Imachi, N. Satoh, H.
Miyake, N. Takido, N. Iwai and A. Kouno: New
hybrid method of total artificial heart replacementwith natural beating by coronary circulatory preservation, Proceedings of 6th European Society for
Artificial Organs, A1-A6, 1979.
7. K. Imachi, I. Fujimasa, H. Miyake, N. Takido, M.
Nakajima, K. Motomura, A. Kouno, T. Ono, and K.
Atsumi:Evaliation of antithrombogenicity, durability
and biocompatibility of an artificial heart system for
more than 100 days, Artificial Organs, 5(Suppl), 423429, 1981.
8. K. Imachi, I. Fujimasa, H. Miyake, N. Takido, M.
Nakajima, A. Kouno, T. Ono, Y. Mori, S. Nagaoka, S.
Kawase, T. Kikuchi, and K. Atsumi: Evaluation ofpolyurethane sac type blood pump after 232 and 288
days total artificial heart pumping without anticoagulant, Trans Am. Soc. Artif. Intern Organs, 27, 118122, 1981
9. K. Atsumi, I. Fujimasa, K. Imachi, H. Myake, N.
Takido, M. Nakajima, A. Kouno, T. Ono, S. Yuasa, Y.
Mori, S. Nagaoka, S. Kawase and T. Kikuchi: Threegoats survived for 288 days, 243 days and 232 days
with hybrid total artificial heart, Trans. Am. Soc.
Artif. Intern. Organs 27, 77- 82 (1981)
10. A. Ihara, K. Motomura, M. Nakajima, K. Imachi, I.
Fujimasa, and K. Atsumi: Comparative study on
Medical laser system and their evaluation criteria,
LASER-TOKYO’81(K. Atsumi et al Ed.), The Jap Soc
for Laser Med, Tokyo, 22-25, 1981.
11. K. Atsumi, I. Fujimasa, K. Imachi, A. Miyamoto, N.
Takido, M. Nakajima S. Tsukagosi and N. Inou:
Development and evaluation on cardiac ventricularassist device for clinical use, Proceedings of ESAO, 9,231- 234, 1982.
12. Fujimasa, K, Imachi, H. Miyake, M. Iwatani and K.
Atsumi: Thermo-blood flowmetry, microprocessor
processed thermograms, Lecture Notes in Medical
Informatics(Lindberg & Leihertz Ed.), SpringerVerlag, 222-227, 1982.
13. K. Imachi, I. Fujimasa, M. Nakajima, A. Miyamoto,
N. Takido, N. Inou., S. Tsukagoshi, K. Mabuchi, K.
Motomura, A. Kouno, T. Ono, and K. Atsumi:
Evaluation of the metabolic state in a total artificial
heart circulatory system, Progress in Artificial
Organs-1983(Atsumi K et al Ed.), 252-256, ISAO
Press, Cleveland, 1984
14. K. Imachi, I. Fujimasa, M. Nakajima, K. Mabuchi, S.
Tsukagoshi, K. Mtomura, A. Miyamoto, N. Takido,
N. Inou, A. Kouno, T. Ono, and K. Atsumi: Overall
analysis of the causes of pathophysiological problems in total artificial heart in animals by cardiac
receptor hypothesis, Trans. Am. Soc. Artif. Intern.
Organs, 30, 591-596, 1984
15. K. Atsumi, I. Fujimasa, K. Imachi and N. Nakajima:
How can the total artificial heart patient be mobileand enjoy his life with an air driven system?, Trans.
Am. Soc. Artif. Intern. Organs, 30, 86-90, 1984
16. N. Inou, I. Fujimasa, K. Imachi, A. Miyamoto, M.
Nakajima, N. Takido, K. Mabuchi, S. Tsukagoshi, A.

Kouno and K. Atsumi: Quantitative approach todetermine the application time of the left ventricularassist device, Artificial Organs, 8(4), 458-469, 1984
17. Fujimasa, K. Imachi, Y. Kajiyama, K. Atsumi: Cardiac
muscle thermo- graphy, Recent Advances in Medical
Thermology(Francic, Ring & Phillips Ed.), Plenum
Publishing, 375-378, 1984
18. K. Atsumi, I. Fujimasa, K. Imachi, M. Nakajima, S.
Tsukagoshi, K. Mabuchi K. Motomura, A. Kouno, T.
Ono, A. Miyamoto, N.Takido and N. Inou: Longterm heart substitution with an artificial heart ingoats, asaio J., 8(3) 155-165,1985
19. Fujimasa, K. Imachi, M. Nakajima, K. Mabuchi, S.
Tsukagoshim A. Kouno T. Ono, N. Takido, K.
Motomura, T. Chinzei, Y. Abe, and K. Atsumi:
Pathophysiological study of a total artificial heartgoat survived for 344 days, Progress in Artificial
Organs-1985(Y. Nose Ed.),ISAO Press, Cleveland,345-353, 1986
20. Y. Abe, M. Nakajima, Y. Orime, K. Maeda, H. Hata,
S. Hosaka, K. Mabuchi T. Chinzei, M. Asano, I.
Fujimasa, K. Imachi, A. Kouno, T. Ono, and K.
Atsumi: Design of contact end of laser probe for contact irradiation laser angioplasty, Nd-YAG laser inMedicine and Surgery(Y. Oguro et al Ed) Professional
Postgraduate Service K.K.,329-332, 1986
21. K. Imachi, I. Fujimasa, K. Mabuchi, T. Chinzei, Y.
Abe, K. Maeda, K. Imanishi, A. Kouno, T. Ono, andK. Atsumi: A newly designed jellyfish valve for an
artificial heart blood pump, Trans. Am. Soc. Artif.
Intern. Organs, 34(3), 726-728, 1988
22. K. Maeda, T. Chinzei, K. Imachi, K. Mabuchi, Y. Abe,T. Yonezawa K. Imanishi, I. Fujimasa, and K.
Atsumi:Predictive control by physical activity rate ofa total artificial heart during exercise, Trans Am Soc
Artif Inter Organs, 34(3),480-484, 1988
23. K. Mabuchi, M. Nakajima, K. Imachi, I. Fujimasa, Y.
Abe, T. Chinzei K. Maeda, K. Imanishi, M. Asano, T.
Yonezawa, and K. Atsumi: Is atrial natriuretic
polypeptide a cause of pathophysiology in total artificial heart animals?, Trans Am Soc Artif Intern
Organs,34(3),463-465, 1988
24. Y. Abe, T. Chinzei, I. Fujimasa, K. Imachi, K.
Mabuchi, K. Maeda, M. Asano A. Kouno, T. Ono,and K. Atsumi: Development of transcutaneous energy transmission system for totally implantable artificial heart, Artificial Heart 2(T. Akutsu Ed.),SpringerVerlag, Tokyo, 257-261, 1988
25. T. Yonezawa, T. Onomura, Y. Abe, K. Mabuchi, T.
Chinzei, K. Imachi, and K. Atsumi: Comparative
study on excimer,CO2 and Nd-YAG laser for biological tissue cutting, Laser Optoelectronics in
Medicine(R. Waidlich & W.Waidlich Ed), SpringerVerlag,31-34, 1988
26. Y. Abe, T. Chinzei, K. Imanishi, T. Yonezawa, S.
Niwa, K. Yotsuya, I. Sogawa, T. Uemiya, S.
Kanazawa, I. Fujimasa, K. Imachi, M. Suzukawa, K.
Mabuchi, M. Asano, Y. Tago, A. Kouno, T. Ono, andK. Atsumi: Laser angioplasty with excimer laser-animal experiment, SPIE,1066,115-122, 1989

27. T. Yonezawa, T. Onomura, R. Kosaka, Y. Miyaji, S.
Tanaka, H. Watanabe, Y. Abe, K. Imachi,ò K.
Atsumi, T. Chinzei, K. Mabuchi, and I. Fujimasa: The
system and procedures of percutaneous intradiscal
laser nucleotomy, SPINE, 15(11), 1175-1185(, 1990
28. K. Imachi, T. Chinzei, Y. Abe, K. Mabuchi, K.
Imanishi, T. Yonezawa, A. Suzuki, T. Isoyama, K.
Atsumi, and I. Fujimasa: Development of flow-transformed pulsatile total artificial heart for total implantation, Artificial Organs, 16(3),308-313, 1992
29. K. Imachi, K. Mabuchi, T. Chinzei, Y. Abe, K.
Imanishi, T. Yonezawa, A. Kouno, T. Ono, H.
Nozawa, T. Isoyama, K. Atsumi, and I. Fujimasa:
Fabrication of a jellyfish valve for use in an artificial
heart, ASAIO Journal, 38(3), 237-242, 1992
30. K. Imachi, T. Chinzei, Y. Abe, T. Isoyama, K.
Mabuchi, K. Imanishi, T. Ono, A. Kouno, M.
Kusakabe, K. Atsumi and I. Fujimasa: A new apparatus for chronic observation of microcirculation in-situ
to evaluate an artificial organ performance. ASAIO
Journal, 40(3) M757-761, 1994
31. K. Imachi, Y.Abe, T.Chinzei, K.Mabuchi, K.Imanishi,
T.Isoyama, A.Kouno, T.ono and I.Fujimasa: Factors
Influencing hemodynamics blood chemical data andhormone secretion of total artificial heart goat. In
Progress in Microcirculation Research(H. Niimi, M.
Oda, T. Sawada, R-J Xiu ed), Pergamon Press,
London, 453-458, 1994
32. Y. Abe, T. Chinzei, K. Imachi, K. Mabuchi, K.
Imanishi, T. Isoyama, H. Matsuura, G. Senih, H.
Nozawa, A. Kouno, T. Ono, K. Atsumi, and I.
Fujimasa: Can total artificial heart (TAH) animals
control their TAH by themselves?- One year survivalof a TAH goat with a new automatic control method.
ASAIO J, 40(3) M506-M509, 1994.
33. T. Isoyama., K. Imachi, Y. Abe, T. Chinzei, K.
Imanishi, K. Mabuchi, H. Matuura, I. Fujimasa: New
Version of flow transformed pulsatile total artificial
heart having no electrical switching valve. Artificial
Organs, 19(7), 694-696, 1995.
34. K. Imachi: Implant science of artificial organs: Design
for success. Artificial Organs, 21(11), 1211-1216,
1997.
35. R. Kosaka, T. Onomura, T. Yonezawa, Y. Ichimura,and K. Imachi: Percutaneous laser nucleotomy for
lumbar disc herniation: Preliminary report of early
results and MRI findings, J Orthop Sci, 1, 102-112,
1996.
36. Y. Abe, T. Chinzei, K. Mabuchi, T. Isoyama, K. Baba,
H. Matsuura, A. Kouno, T. Ono, S. Mochizuki, Y.
Sun, K. Imanishi, K. Atsumi, I. Fujimasa and K.
Imachi: Over 500 days’ survival of a goat with a total
artificial heart with 1 / R control. In Heart
Replacement-Artificial Heart 6(ed. T. Akutsu, H.
Koyanagi), Springer-Verlag, Tokyo, 34-38, 1998.
37. K. Mabuchi, Y. Abe, S. Shimizu, K. Imachi, T.
Chinzei, H. Matsuura, T. Isoyama, K. Baba, K.
Imanishi, S. Mochizuki, Y. Sun, T. Tago, A. Kouno,
T. Ono, and I. Fujimasa: Pathological study of a goatthat survived for 532 days with a total artificial heart109

using the 1 / R control method. In Heart
Replacement-Artificial Heart 6(ed. T. Akutsu, H.
Koyanagi), Springer-Verlag, Tokyo, 313-317, 1998.
38. Y. Abe, T. Chinzei, K. Mabuchi, A. J. Snyder, T.
Isoyama, K. Imanishi, T. Yonezawa, H. Matsuura, A.
Kouno, T. Ono, K. Atsumi, I. Fujimasa, and K.
Imachi: Physiological control of a total artificial
heart-Conductance and arterial pressure based control, J. Appl. Physiol., 84(3), 868-876, 1998.
39. Y. Abe, T. Chinzei, T. Ono, S. Mochizuki, I. Saito, P.
Guba, T. Karita, Y. P. Sun, A. Kouno, T. Isoyama, K.
Baba, and K. Imachi: Implantation of the undulation
pump total artificial heart in the goat, Artificial
Organs, 23(10), 932-938, 1999.
40. K. Imachi, T. Chinzei, Y. Abe, T. Isoyama, S.
Mochizuki, I. Saito, T. Ono, T. Karita, H. Matsuura,
A. Kouno, K. Mabuchi, I. Fuhimasa, and K. Atsumi:
Implantable total artificial heart - History and present status at the University of Tokyo, J. of Artificial
Organs, 2(1): 13-23,
41. T. Karita, K. Imachi, T. Taguchi, M. Akashi, K. Sato,and J. Tanaka: In vitro calcification model Part -1:
Apatite formation on segmented polyurethane containing silicone using an alternate soaking process. J
of Bioactive and Compatible Polymers, 15, 72-84,
2000
42. S. Mochizuki, Y. Abe, T. Chinzei, T. Isoyama, T.
Ono, I. Saito, P. Guba, T. Karita, Y. P. Sun, A. Kouno,
T. Suzuki, K. Baba, K. Mabuchi, and K. Imachi:
Results of animal experiments using an undulation
pump total artificial heart: Analysis of 10 day and 19
day survival, ASAIO Journal, 46(4): 500-504, 2000
43. K. Imachi, T. Chinzei, Y. Abe, K. Mabuchi, H.
Matsuura, T. Karita, K. Iwasaki, S. Mochizuki, Y-P
Son, I. Saito, A. Kouno, and T. Ono: A new hypothesis on the mechanism of calcification formed on a
blood contacted polymer surface. J. of Artificial
Organs, 4(1), 74-82, 2001.
44. Y. Abe, T. Chinzei, T. Isoyama, T. Ono, S.
Mochizuki, I. Saito, K. Iwasaki, M. Ishimaru, T.
Karita, A. Kouno, K. Baba, and K. Imachi: One monthsurvival with undulation pump total artificial heart
in a goat, Artificial Organs, 25(1): 69-71, 2001.
45. Y. Abe, T. Chinzei, T. Isoyama, T. Ono, S.
Mochizuki, I. Saito, K. Iwasaki, M. Ishimaru, A.
Baba, A. Kouno, T. Karita, K. Baba, and K. Imachi:
Progress in the development of the undulation pumptotal artificial heart, J. of Congestive Heart Failure
and Circulatory Support, 1(4), 167-170, 2001, and In
Heart Replacement-Artificial Heart 7(ed. T. Akutsu,
H. Koyanagi), ISIS Medical Media Ltd. England, 167170, 2001.
46. Saito, Y. Abe, T. Chinzei, T. Ono, T. Isoyama, S.
Mochizuki, K. Iwasaki, A. Baba, A. Kouno, T. Karita,
M. Ishimaru, A. Baba, and K. Imachi: Miniature
undulation pump for the study of renal circulation, J.
of Congestive Heart Failure and Circulatory Support,
1(4), 321-325, 2001, and In Heart ReplacementArtificial Heart 7(ed. T. Akutsu, H. Koyanagi), ISIS
Medical Media Ltd. England, 321-325, 2001.

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47. Y. Abe, T. Chinzei, T. Isoyama, T. Ono, T. Ozeki, I.
Saito, A. Kouno, and K. Imachi: Principle of therotary undulation pump, J Artif Organs, 5(2), 84-90,
2002.
48. K. Iwasaki, M. Umezu, K. Iwasaki, Y.Abe, T.Chinzei,
T.Isoyama, I.Saito, M. Ishimaru, K. Imachi: The
improved jellyfish valve: Durability, enhancementwith sufficient blood compatibility, ASAIO Journal,
48: 532 - 537, 2002.

Department of
Neuropathology
Outline and Research Objectives
The long-term goal of the research in our department is to determine the molecular
events that lead to the development of Alzheimer’s disease (AD). For this purpose, we are
analyzing two proteins that have been implicated in the AD pathogenesis: amyloid ß-protein (Aß) and tau.

Faculties and Students
Professor and Chair
Lecturer
Associate

Yasuo Ihara, M.D. (1991~)
Maho Morishima, Ph.D.
Satoru Funamoto, Ph.D.
Kozo Motonaga, Ph.D.
Graduate Student....................7
Secretary .................................1

Past Research and Major Accomplishments
Identification of posttranslational modificationsof tau in paired helical filaments
Tau in paired helical filaments (PHF) is characterized by hyperphosphorylation and ubiquitination. The
former causes the tau to migrate slower that dephosphorylated counterparts, and such particular tau
makes three distinct bands on the Western blot, andcalled PHF-tau. We found that this phosphorylation
somehow resemble that seen in fetal tau, because
many phosphorylation-dependent monoclonal antibodies that react with PHF-tau recognize fetal tau.
This indicates that fetal tau is phosphorylated to agreater extent than adult tau. We underwent thework to determine the exact sites for phosphorylation, using selective modification of phosphorylated
Ser residues and their identification of amino acidsequencing and mass spec. As a result more than 20
phosphorylation sites in PHF-tau were detemined:
about half number of the sites were proline-directed,
while the remaining were nonproline-derected. By ourwork the phosphorylation of PHF-tau was fully characterized.
PHF was found to be ubiquitinated in 1987, andsince then we attempted to determine the ubiquitibation sites in tau, but failed because of purification ofubiquitinated tau was very difficult and provided only
very poor yield. From smeared tau (see below) we
obtained a small amounts of purified ubiquitinatedtau, which was digested by lysysl endopeptidase andgenerated fragments were subjected to mass spec.
Thus ubiquitinated sites were accurately determined,

and found in the microtubule-binding domain of tau.
One of the most remarkable characteristics of thetau in PHF is smearing on the blot from high molecular mass region down to 1-2 kD. The extent of smearing is well-correlated with the density of neurofibrillary tangles. We purified smeared tau using HPLC andanalyzed protein chemically. Deamidation and isoaspartate formation were found, suggesting thatsmeared tau represents aging of NFT in vivo.
(2) Determination of Aß42 as the predominant
species deposited in the brain
At that time, two Aß species were known, but
their significance was unknown, although in the culture media Aß40 that terminates in Val-40 is the
major species. Using end-specific monoclonals thatwere developed by N. Suzuki, we found immnocytochemically that Aß42 that terminates in Ala-42, aminor species among the secreted Aß, is predominant
in the parenchyma of AD cortex. To learn about thetemporal profile for Aß deposition in the humanbrain, we examined the brains from Down syndrome
patients who invariably develop Alzheimer pathologies after the middle ages. In this way we found thatthe first species deposited in the brain is Aß42.
(3) Quantification of Aß accumulation in thehuman brain
With collaboration with N. Suzuki, we constructed
two-site ELISA highly specific for Aß40 or Aß40. We
first applied the ELISA for the quantification of Aß40
and Aß42 in the human brain. The biochemical alterations specific to AD may start much earlier than we
previously thought. The work on CAA suggests fourstages in the evolution: biochemically detectablestage, immunocytochemically detectable stage, histologically (Congo red) detectable stage, and clinically(multiple and recurrent hemorrhage in the aged)
detectable stage. In other words, clinically overt
patients are just a tip of the iceberg. The similar timecourse can be applicable to parenchymal Aß deposition; biochemical accumulation of Aß should start in
the 40s (or even before). And AD patients are most111

likely to represent a tip of the iceberg. A straightforward interpretation would be that AD is the consequence of the longstanding biochemical abnormalities.
Here, we focus on the nature of the initial biochemical abnormality, eventually leading to ß-amyloidogenesis, rather than on why Aß accumulate in
the brain in an exponential way. In particular, we are
currently concerned about possible roles of low-density membrane domain (detergent-insoluble, glicolipid
and cholesterol enriched domain; DIGs) in the Aß 40
and Aß42 metabolism, and in the earliest stage of ßamyloidogenesis.
As Aß accumulates, seemingly, Aß42 shifts tohigher density fractions, but Aß40 tends to stay low.
Possible explanations include: When a large amountof Aß accumulated, the low-density membrane
domain may shift to higher density or Aß42-boundlow-density membrane domain may lose specific lipid
composition (cholesterol and/or sphingolipid), resulting in shift to higher density. In either way, there may
be a certain critical level of Aß42 in the domainbeyond which the membrane-bound Aß42 cannot
float.
(4) Accelerated Aß accumulation in ε4-bearing
subjects
We attempted to determine when Aß deposition
starts in most human brains and how it is influenced
by apolipoprotein E (apoE) allele ε4, a strong risk factor for late-onset AD. We successfully quantitated,using an improved extraction protocol and sensitive
ELISA, the Aß40 and Aß42 levels in the insoluble
fraction of the brains from many nondemented subjects aged 22 to 81 years. Both Aß40 and Aß42 weredetectable in the insoluble fraction of the brains evenfrom young subjects aged 20 to 30 years. The incidence of the subjects significantly accumulating Aßincreased in an age-dependent manner; Aß42 levels
arose steeply in some subjects in their late 40s andthis was accompanied by a much smaller rise in Aß40
levels. ApoE ε4 was found to significantly enhance
Aß42 and, to a less extent, Aß40 accumulation. These
results strongly suggest that the presence of ε4 allele
sets earlier in life the start of Aß accumulation in thebrain.
(5) Discovery of ε-cleavage using a cell-free system
Aß is generated from ß-amyloid precursor protein(APP) when ß-secretase cleavage at the extracellulardomain produces a 99-residue C-terminal fragment
called CTF99 or CTFß. Subsequent cleavage of CTFßin the middle of the transmembrane domain by γ-secretase primarily produces either a 40-residue protein(Aß40) or a 42-residue protein (Aß42). Intramembrane

112

γ-secretase cleavage of CTFß should yield a 59- or 57residue cytoplasmic C-terminal fragment (CTFγ41-99
or 43-99; APP712-770 or 714-770 according to thenumbering of APP770 isoform) called CTFγ. To learn
more about the properties of γ-cleavage, we undertook
to establish cell-free system for generation of Aß and
CTFγ, and characterize CTFγ, the other product of thisunusual cleavage. The generated CTFγ consists largelyof CTFγ50-99, and of CTFγ49-99, although the latter isseveral to ten-fold less than the former. Comparisonof the cleavage sites of APP, APLP1, and APLP2 identifies new cleavage site: two to five residues inside thecytoplasmic membrane boundary. Therefore, thisintramembrane cleavage is distinct from the Aß-generating γ-cleavage that occurs in the middle of thetransmembrane domain (more than nine residues
inside the membrane boundary).
It has been unclear why APP is cleaved near the
middle of the transmembrane domain, whereas Notch
1 is cleaved immediately inside the cytoplasmic membrane boundary. Overall, our findings strongly suggestthat the cleavage of Notch corresponds to this distinct
type of cleavage rather than to γ–cleavage.

Current Research and Future Prospects(1) Characterization of the substrate specificity ofγ-secretase
1. Is ε-cleavage a reality? Can we exclude the possibility that γ-cleavage is followed by swift, stepwise cleavages by a particular amino peptidase?
Definite evidence requires demonstration of
longer Aß (Aß1-49 or 1-50) and/or the middle segment (Aß41-49). Can one detect a longer Aßspecies in the membrane?
2. If it is reality, what is the meaning of γ-cleavage?
Does the presence ragged C termini of Aß suggest
participation of a particular carboxyl peptidase in
the generation of Aß?
3. If so, what is the relationship between γ- and εcleavage? The two cleavages may not be in one to
one stoichiometric relationship.
4. The general principle of type 1 membrane protein
degradation. shedding, release of ectodomain, followed by intramembrane (ε-) cleavage. How is theintramembrane segment left after ε-cleavage
degraded?
5. Is Aß40 and Aß42 production separable? How is
Aß40 or Aß42 generation correlated to CTFγ49-99
or CTFγ50-99?
(2) Mechanism of Aß accumulation in the brain
1. At present the mechanism of FAD (Familial AD)
can be understood by altered enzyme-substraterelationship: Mutations in enzyme (presenilin) or
substrate (APP) causes an (small) increase in the
Aß42 production, which eventually leads to AD.
Patients with presenilin or APP mutants usuallyshow higher levels of Aß42 in the plasma.
Presumably, whether this relationship is altered
can be detected by increased levels of Aß42 in theplasma. Another pathway to development of AD
is being proposed: increased tendency to protofibril formation.
2. The role of ApoE4 is not to modify the enzymesubstrate relationship, but to mediate throughother pathways, thereby leading to earlier buildup of senile plaques. Potential target would be in
the exchange of Aß between cell membrane and
HDL particles, in the transport of Aß through BBB
or in the Aß fibrillization.
3. The thought into exponential accumulation curve
may provide us with some insights into the mechanism. The accumulation curve may be simulatedby the equation: A(t)=C(t-40)n/1+(t-40)n, where C:
constant, t, age>40; steepness in the accumulation
depends on n. This equation represents cooperativity phenomena (phase transition), and may suggest that age-dependent Aß accumulation may be
one of such examples in the biology. If so, theunderlying mechanism can be explained byenhanced Aß aggregation force. A point is why
the aggregation potential is enhanced. One possibility is a decrease in the degradation potential(decreased protease activity) during aging.
However, it is a bit difficult to envision, because it
has not so far been known that certain proteases
are acutely inactivated during aging. The otherpossibility is altered environments induced byaging. Altered lipid composition and/or generationof a certain factor to promote Aß aggregation.
Altogether, accumulation is a consequence of
aggregation, and aggregation may be a consequence of an alteration in the environment, but
may not be decreased protease activity. In thisregard, it may be important to investigate the significance of Aß accumulation in the raft. First, toexamine whether raft-accumulated Aß is younger
than extracelluarly deposited Aß by using
pyroGlu-specific antibodies. Other markers for invivo aging include isomerization and racemizationof Asp residues and truncation of the Aß molecule.
(3) Tau and neuronal cell death
What do we know about the significance of intracellular tau deposition, and what should we do in the
future?
1. NFT itself is not so harmful to cell. NFT-bearing
neurons are viable, and still attempt to maintain

2.

3.

4.

5.

6.

7.

dendritic arborization. NFT-bearing oligodendrocytes rather specific for FTDP-intron mutations
still supply enough amounts of myelin. No
demyelination is detected by a classical method.
But often, dendrites of NFT-bearing neurons arecompromised, which reminds us of dying-backneuropathy. Presumably, curly fibers represent
dying-back segments of basal dendrites isolated
from neuronal cell bodies.
FTDP-17 is characterized by tauopathy and neuronal loss. P301L mutation is featured by extensive neuronal loss rather than NFT formation (seetransgenic fly; Wittmann CW et al. Science
2001;293:711-4), while R406W mutation is featured by NFT formation rather than neuronal loss.
Clinical course may depend on neuronal loss
rather than NFT formation. This is consistent
with the old view that the degree of dementia in
AD is best correlated with the extent of neuronal
loss.
Thus, the most important question about tau is
why and how tau kills the neuron, rather than
why and how it aggregates into NFT.
Significance of hyperphosphorylation of tau in
PHF or PHF-like fibrils. Hyperphosphorylation of
tau may be a consequence of degeneration of neuron, but not a cause of degeneration. Study on
R406W brain showed that i) cytosolic R406W tau
is less phosphorylated, which is consistent withan observation with CHO stable transfectant; ii)
but the mutant tau composing PHF-like fibrils is
hyperphosphorylated. A most likely interpretation
is as follows. The mutant tau has a bulky residue
close to its microtubule-binding domain, andwhen the mutant tau interact with tubulin or
microtubules, a particular kinase cannot access toSer-396 and 404, resulting in hypophosphorylation on these residues. Once the interaction withtubulin or microtubules is lost (by degeneration),
the mutant tau takes a random structure as does
wild-type tau, and these residues are freely
accessed by kinase and become hyperphosphorylated.
To do: establishment of FTDP tau knock-in mice;
attempt to increase the tau expression level by
removing the neo gene by breeding with CAG Cre
mice. Breeding with APP transgenic mice (forexample, APPsw mice) may make the pathologicalphenotype to appear within lifespan, and P301L
mice may develop NFT and neuronal loss muchearlier than wild-type knock-in mice.
More data about tau in the cell biology may provide some keys to understanding of the pathogenesis of FTDP-17. For example, differential role ofthree-repeat and four-repeat tau should be more
highlighted.

113

8. The data from transgenic fly suggest that expression of even wild-type tau may be somehow
harmful to viability of fly (Wittmann CW et al.
Science. 2001;293:711-4). This can be applied tosynuclein also (Feany MB, Bender WW. Nature
2000;404:394-8). Presumably, the essential featureof FTDP-17 is neuronal death without overt NFT
formation. But in many FTDP-17 cases, both neuronal loss and NFT formation coexist. This meansthat the initial step to neuronal death may beshared by the pathway to NFT formation. In thisrespect, recent observations may be significant(Zhukareva V et al. Loss of brain tau defines novel
sporadic and familial tauopathies with frontotemporal dementia. Ann Neurol. 2001;49:165-75).
9. Currently there is no unifying hypothesis that can
explain remarkable heterogeneity of FTDP-17. If it
does exist, it must explain i) Why exonic mutation cause neuronal degeneration as well as ii)
Why increased levels of four-repeat tau cause neuronal degeneration.

Research Grants
1. Core Research for Evolutional Science andTechnology from the Japan Science and Technology
Corporation
Year : 1996 ~ 2001
¥483,000,000.2. Grant-in-Aid for Scientific Research on Priority
Areas, Advanced Brain
Science Project, from the Ministry of Education,
Culture, Sports, Science and Technology, Japan
Year : 2000 ~ 2002
¥108,000,000.3. Research Grants for Longevity Sciences from the
Ministry of Health and Welfare, Japan
Year: 1999 ~ 2001
¥ 6,000,000.4. Grant-in-Aid for Scientific Research (C) from the
Ministry of Education, Culture, Sports, Science andTechnology, Japan
Year: 1999 ~ 2000
¥ 3,700,000.5. Research Grants for Longevity Sciences from the
Sasakawa Health Science Foundation, Japan
Year: 1999
¥ 2,900,000.-

Select Publications
1. Oyama F, Shimada H, Oyama R, Titani K, Ihara Y:
Differential expression of ß amyloid protein precursor (APP) and tau mRNA in the aged human brain:
Individual variability and correlation between APP751 and four-repeat tau. J Neuropathol Exp Neurol
50: 560-578, 1991

114

2. Uchida Y, Takio K, Titani K, Ihara Y, Tomonaga M:
The growth-inhibitory factor that is deficient in the
Alzheimer’s disease brain is a 68-amino acid metallothionein-like protein. Neuron 7: 337-347, 1991
3. Watanabe N, Takio K, Hasegawa M, Arai T, Titani K,
Ihara Y: Tau 2: A probe for a Ser-conformation in theamino terminus of t. J Neurochem 58: 960-966, 1992
4. Kanemaru K, Takio K, Miura R, Titani K, Ihara Y:
Fetal-type phosphorylation of the tau in paired helical filaments. J Neurochem 58: 1667-1675, 1992
5. Iwatsubo T, Hasegawa M, Esaki Y, Ihara Y: Lack ofubiquitin immunoreactivities at both ends of neuropil threads —— possible bidirectional growth ofneuropil threads ——. Am J Pathol 140: 277- 282,
1992
6. Hasegawa M, Morishima-Kawashima M, Takio K,
Suzuki M, Titani K, Ihara Y: Protein sequence andmass spectrometric analyses of tau in the
Alzheimer’s disease brain. J Biol Chem 267: 1704717054, 1992
7. Oyama F, Shimoda H, Oyama R, Titani K, Ihara Y: A
novel correlation between the levels of ß-amyloid
protein precursor and tau transcripts in the aged
human brain. J Neurochem 59: 1117-1125, 1992
8. Tsuji S, Kobayashi H, Uchida Y, Ihara Y, Miyatake T:
Molecular cloning of human growth inhibitory factor
cDNA and its down-regulation in Alzheimer’s disease. EMBO J 11:4843-4850, 1992
9. Oyama F, Shimada H, Oyama R, Titani K, Ihara Y: ßAmyloid protein precursor and tau mRNA levels versus ß-amyloid plaque and neurofibrillary tangles in
the aged human brain. J Neurochem 60:1658-1664,
1993
10. Hasegawa M, Watanabe A, Takio K, Suzuki M, Arai
T, Titani K, Ihara Y: Characterization of two distinct
monoclonal antibodies to paired helical filaments(PHF) — Further evidence for fetal-type phosphorylation of the tau in PHF —. J Neurochem 60:20682077, 1993
11. Morishima-Kawashima M, Hasegawa M, Takio K,
Suzuki M, Titani K, Ihara Y: Ubiquitin is conjugatedwith amino-terminally processed tau in paired helicalfilaments. Neuron 10:1151-1160, 1993
12. Yoshida H, Ihara Y: t in paired helical filaments isfunctionally distinct from fetal t. — assembly incompetence of paired helical filament-t —. J Neurochem
61:1183-1186, 1993
13. Watanabe A, Hasegawa M, Suzuki M, Takio K,
Morishima-Kawashima M, Titani K, Arai T, Kosik
KS, Ihara Y: In vivo phosphorylation sites in fetal
and adult rat tau. J Biol Chem 268:25712-25717,
1993
14. Uéda K, Fukushima H, Masliah E, Xia Y, Iwai A,
Yoshimoto M, Otero DAC, Kondo J, Ihara Y, Saitoh
T: Molecular cloning of a novel component of amyloid in Alzheimer’s disease. Proc Natl Acad Sci USA
90:11282-11286, 1993
15. Oyama F, Cairns NJ, Shimada H, Oyama R, Titani K,
Ihara Y: Down’s syndrome: Upregulation of ß-amyloid protein precursor and tau mRNAs and theirdefective coordination. J Neurochem 62:1062-1066,
1994
16. Iwatsubo T, Hasegawa M, Ihara Y: Neuronal andglial tau-positive inclusions in diverse neurologic diseases share common phosphorylation characteristics.
Acta Neuropathol 88:129-136, 1994
17. Iwatsubo T, Odaka A, Suzuki N, Mizusawa H,
Nukina N, Ihara Y: Visualization of Aß42(43) and
Aß40 in senile plaques with end-specific Aß-monoclonals: Evidence that an initially deposited species is
Aß42(43). Neuron 13:45-53, 1994
18. Suzuki N, Iwatsubo T, Odaka A, Ishibashi Y, Kitada
C, Ihara Y: High tissue content of soluble ß1-40 islinked to cerebral amyloid angiopathy. Am J Pathol
145:452-460, 1994
19. Oyama F, Oyama R, Ihara Y: Apolipoprotein E
Genotype versus Alzheimer’s pathologies and related-gene expression in the aged population. Mol Brain
Res 29:92-98, 1995
20. Morishima-Kawashima M, Hasegawa M, Takio K,
Suzuki M, Titani K, Ihara Y: Proline-directed andnon-proline-directed phosphorylation of PHF-tau. J
Biol Chem 270:823-829, 1995
21. Iwatsubo T, Mann DMA, Odaka A, Suzuki N, Ihara
Y: Amyloid ß-protein (Aß) deposition: Aß42(43) precedes Aß40 in Down’s syndrome. Ann Neurol
37:294-299, 1995
22. Saido TC, Iwatsubo T, Mann DMA, Shimada H,
Ihara Y, Kawashima S: Dominant and differential
deposition of distinct ß-amyloid peptide species,
AßN3(pE), in senile plaques. Neuron 14:457-466,
1995
23. Uchida Y, Ihara Y: The N-terminal portion of growth
inhibitory factor is sufficient for biological activity. J
Biol Chem 270: 3365-3369, 1995
24. Shinkai Y, Yoshimura M, Ito Y, Odaka A, Suzuki N,
Yanagisawa K, Ihara Y: Amyloid ß-proteins (Aß)1-40
and 1-42(43) in the soluble fraction of extra- andintracranial blood vessels. Ann Neurol 38:421-428,
1995
25. Yanagisawa K, Odaka A, Suzuki N, Ihara Y: GM1
ganglioside-bound amyloid ß-protein (Aß): A possible
form of preamyloid in Alzheimer’s disease. Nature
Med 1: 1062-1066, 1995
26. Iwatsubo T, Yamaguchi H, Fujimuro M, Yokosawa
H, Ihara Y, Trojanowski JQ, Lee VM-Y: Purificationand characterization of Lewy bodies from the brainsof patients with diffuse Lewy body disease. Am J
Pathol 148:1517-1529, 1996
27. Gu Y, Oyama F, Ihara Y: t is widely expressed in rat
tissues. J Neurochem 67:1235-1244, 1996
28. Namekata K, Oyama F, Imagawa M, Ihara Y: Humantransferrin (Tf) : A single mutation at codon 570
determines the TfC1 or TfC2 variant. Human Genet
100: 457-458, 1997
29. Yamazaki T, Haass C, Saido TC, Omura S, Ihara Y:
Specific increase in amyloid ß-protein 42 secretion
ratio by calpain inhibition. Biochemistry 36: 83778383, 1997

30. Hamano T, Yoshimura M, Yamazaki T, Shinkai Y,
Yanagisawa K, Kuriyama M, Ihara Y: Amyloid ß-protein (Aß) accumulation in the leptomeninges during
aging and in Alzheimer disease. J Neuropathol Exp
Neurol 56: 922-932, 1997
31. Wild-Bode C, Yamazaki T, Capell A, Leimer U,
Steiner H, Ihara Y, Haass C: Intracellular generation
and accumulation of amyloid ß-peptide terminating
at amino acid 42. J Biol Chem 272: 16085-16088,
1997
32. Shinkai Y, Yoshimura M, Morishima-Kawashima M,
Ito Y, Shimada H, Yanagisawa K, Ihara Y: Amyloid ßprotein deposition in the leptomeninges and cerebral
cortex. Ann Neurol 42: 899-908, 1997
33. Funato H, Yoshimura M, Yamazaki T, Saido TC, Ito
Y, Yokofujita J, Okeda R, Ihara Y: Astrocytes containing amyloid ß-protein (Aß)-positive granules are associated with Aß40-positive diffuse plaques in the aged
human brain. Am J Pathol 152: 983-992, 1998
34. Yoshida H, Watanabe A, Ihara Y: Collapsin response
mediator protein (CRMP)-2 is associated with neurofibrillary tangles in Alzheimer’s disease. J Biol
Chem 273: 9761-9768, 1998
35. Nakabayashi J, Yoshimura M, MorishimaKawashima M, Ito Y, Funato H, Miyakawa T,
Yamazaki T, Ihara Y: Amyloid ß-protein (Aß) accumulation in the putamen and mammillary body during aging and in Alzheimer’s disease. J Neuropathol
Exp Neurol 57: 343-352, 1998
36. Oyama F, Sawamura N, Kobayashi K, MorishimaKawashima M, Kuramochi T, Ito M, Tomita T,
Maruyama K, Saido TC, Iwatsubo T, Capell A, Walter
J, Grünberg J, Ueyama Y, Haass C, Ihara Y: Mutant
presenilin 2 transgenic mouse: Increased Aß levels
and a marked age-dependent increase in the Aß42
ratio in the brain. J Neurochem 71: 313-322, 1998
37. Funato H, Yoshimura M, Yamazaki T, Saido TC, Ito
Y, Yokofujita J, Okeda R, Ihara Y: Quantitation ofamyloid beta-protein (A beta) in the cortex during
aging and in Alzheimer’s disease. Am J Pathol 152:
1633-1640, 1998
38. Murakami N, Oyama Y, Gu Y, McLennan IS, Nonaka
I, Ihara Y: Accumulation of tau in autophagic vacuoles in chloroquine myopathy. J Neuropathol Exp
Neurol 57: 664–673, 1998
39. Morishima-Kawashima M, Ihara Y: The presence ofamyloid ß-protein in the detergent-insoluble membrane compartment of human neuroblastoma cells.
Biochemistry 37: 15247-15253, 1998
40. Enya M, Morishima-Kawashima M, Yoshimura M,
Shinkai Y, Kusui K, Khan K, Games D, Schenk D,
Sugihara S, Yamaguchi H, Ihara Y: Appearance of
sodium dodecyl sulfate-stable amyloid ß-protein (Aß)
dimer in the cortex during aging. Am J Pathol 154:
271-279, 1999
41. Watanabe A, Takio K, Ihara Y: Deamidation andisoaspartate formation in smeared tau in paired helical filaments— unusual properties of the microtubule-binding domain of tau —. J Biol Chem 274:7368-7378, 1999115

42. Matsumura N, Yamazaki T, Ihara Y: Stable expression in Chinese hamster ovary cells of mutated tau
genes causing frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17). Am J
Pathol 154: 1649-1656, 1999
43. Funato H, Enya M, Yoshimura M, MorishimaKawashima M, Ihara Y: The presence of sodium
dodecyl sulfate-stable amyloid ß-protein dimers in
the hippocampus CA1 not exhibiting neurofibrillarytangle formation. Am J Pathol 155: 23-28, 1999
44. Misonou H, Morishima-Kawashima M, Ihara Y:
Oxidative stress induces intracellular accumulationof amyloid ß-protein (Aß) in human neuroblastoma
cells. Biochemistry 39:6951-6959, 2000
45. Sawamura N, Morishima-Kawashima M, Waki H,
Kobayashi K, Kuramochi T, Ito M, Oyama F, Ando S,
Ihara Y: Mutant-presenilin 2-transgenic mice: A large
increase in the levels of Aß42 is presumably associated with the low-density membrane domain that contains decreased levels of glycerophospholipids andsphingomyelin. J Biol Chem 275:27901-27908, 2000
46. Morishima-Kawashima M, Oshima N, Ogata H,
Yamaguchi H, Yoshimura M, Sugihara S, lhara Y:
Effect of apolipoprotein E allele ε4 on the initial
phase of amyloid ß -protein accumulation in thehuman brain. Am J Pathol 157: 2093-2099, 2000
47. Oshima N, Morishima-Kawashima M, Yamaguchi H,
Yoshimura M, Sugihara S, Khan K, Games D, Schenk
D, Ihara Y: Accumulation of amyloid beta-protein in
the low-density membrane domain accuratelyreflects the extent of beta-amyloid deposition in thebrain. Am J Pathol 158: 2209-2218, 2001
48. Miyasaka T, Morishima-Kawashima M, Ravid R,
Heutink P, van Swieten JC, Nagashima K, Ihara Y:
Molecular analysis of mutant and wild-type tau
deposited in the brain affected by FTDP-17 R406W
mutation. Am J Pathol 158: 373-379, 2001
49. Yamazaki T, Chang TY, Haass C, Ihara Y:
Accumulation and aggregation of amyloid ß-protein
in late endosomes of Niemann-Pick Type C cells. J
Biol Chem 276: 4454-4460, 2001
50. Gu Y, Misonou H, Sato T, Dohmae N, Takio K, Ihara
Y: Distinct intramembrane cleavage of the ß-amyloid
precursor protein family resembling γ-secretase-likecleavage of Notch. J Biol Chem 276:35235-35238,
2001

116

Department of
Cognitive Neuroscience
Outline and Research Objectives
The Department of Cognitive Neuroscience is one of the three departments composing
the Speech and Cognitive Science Section. Since the establishment of this department in
1991, we have been working in the field of brain and cognition, especially from the point ofview of language.
Main research topics:
1. Research on the brain damaged patient: callosal syndrome, right temporal lobe syndrome, macular sparing, aphasia therapy
2. Functional magnetic resonance imaging (fMRI) during cognitive process: mental writing,writing, face recognition, reading comprehension, naming, reading aloud
3. Magnetoencephalographic (MEG) study of cognitive function: early component of thevisual evoked magnetic field, motor imagery
4. Brain mechanism of Japanese kanji processing
Teaching activities:
1. Graduate course: Speech science and language communication, Pathophysiology ofspeech and language Communication, Introduction to neuroscience
2. Master course: Cognitive neuroscience
3. Graduate course, Faculty of Literature: Experimental phonetics
4. Undergraduate course: Speech and language communication, Medical data processing
5. Undergraduate course, Faculty of Education: Fundamentals of speech science
Clinical activities:
The assessments and therapies for aphasia, apraxia, agnosia, amnesia and dementia are
conducted in collaboration with the Deptment of Neurology, Department of Neurosurgery
and Department of Otorhinolaryngology

Faculties and Students
Professor

Morihiro Sugishita Dr. Health Sci.,
Dr. Medical Sci. 1993~
Associate .................................2
Graduate Students .................2.
Research Students...................1
Guest Researchers...................2

Past Research and Major Accomplishments
1. Research on the Brain Damaged Patient1-1) Sugishita M, et al. Dichotic listening inpatients with partial section of the corpus callosum.
Brain 118, 417-427, 1995 : Despite the common
assumption that damage to the posterior part of thetrunk of the corpus callosum causes strong left-ear
suppression, the results indicated that damage to thesplenium causes strong left-ear suppression.

1-2) Koike A, et al. Preserved musical abilities following right temporal lobectomy. Journal of
Neurosurgery 85, 1000-1004, 1996: No disturbancesin the Seashore Measures were detected after temporal lobectomy on either side.
1-3) Sugishita M, et al. Hemispheric representationof the central retina of comissurotomized subjects.
Neuropsychologia 32, 399-415, 1994: The region of
the right (temporal) hemiretina represented by bothhemispheres in letter processing, if it exists, was estimated as less than 0.6º from the foveal center.
2. Functional Magnetic Resonance Imaging (fMRI)
During Cognitive Process2-1) Sugishita M, et al. Functional maguetic resonance imaging (fMRI) during mental writing withphonograms. NeuroReport 7, 1917-1921, 1996: Four
regions were activated during mental writing in all six
subjects; the left intraparietal sulcus, the middle partof the left precentral sulcus and the posterior part of117

the left superior frontal sulcus, the right intraparietal
sulcus, and either or both of the left and right cingulate sulci. The left intraparietal region was usually themost extensively activated.
2-2) Katanoda K, et al. A functional MRI study on
the neural substrates for writing. Human Brain
Mapping 13, 34-42, 2001: During writing, activations
were observed in the anterior part of the left superiorparietal lobule, the posterior part of the left middle
and superior frontal gyri, and the right cerebellum.
2-3) Katanoda K, et al. Neural substrates for therecognition of newly learned faces: a functional MRI
study. Neuropsychologia 38, 1616-1625, 2000: The
bilateral fusiform gyrus is involved, not only in faceperception, but in a certain aspect of face recognition
memory, and this aspect is related to the actual recognition of previously viewed faces rather than the processing of novel ones. The right parietal and frontal
regions, in contrast, are differentially more associatedwith the detection of novel faces or retrieval effort.
3. Magnetoencephalographic (MEG) Study of
Cognitive Function3-1) Yoneda K, et al. The early component of thevisual evoked magnetic field. NeuroReport 6, 797-800,
1995: The early component was observed in three ofthe nine subjects. The latency ranged from 40 to 45
ms in MEG and from 39 to 47 ms in EEG. The resultof dipole localization analysis showed that its origin
was cortical, and specifically, the striate cortex.
3-2) Ogiso T, et al. The precuneus in motor
imagery: a magnetoencephalographic study.
NeuroReport 11, 1345-1349, 2000: MEG was applied
to subjects who imagined themselves hurdling in selfcentered space. In three of six subjects all 300 trials in
the motor imagery condition revealed the precuneus
dipole. When we divided the 300 trials into four overlapping blocks, all six subjects showed precuneus
activity.
4. Brain Mechanism of Japanese Kanji Processing4-1) Sugishita M, Omura K. Learning Chinese characters may improve visual recall. Perceptual andMotor Skills 93, 579-594, 2001 : From elementarythrough high school, Japanese children are required tomemorize a large number of distinct visual forms, i.e.,roughly 2,000 Chinese characters, and tremendous
effort is expended in learning to read and write them.
We hypothesized that early training in memorizing
Chinese characters and the use of these characters indaily life shapes brain development and facilitatesrecall of visual forms in general. We administered theWechsler Memory Scale-Revised (WMS-R) to a repre-

118

sentative sample of the normal Japanese population(316 persons, 100% Japanese) and compared theirscores with data previously obtained from a representative sample of the normal U.S. population (316 persons, 82.5% Caucasian). Compared to the Americans,
the Japanese group obtained significantly higherscores on these two visual recall subtests in all six age
groups (16 to 74 years old).

Current Research
Research on the brain damaged patients: callosal
syndrome, right temporal lobe syndrome, macularsparing, aphasia therapy
1. fMRI during cognitive process: mental writing,writing, face recognition, reading comprehension,naming, reading aloud2. fMRI study on speech dominance in split-brain
patients
3. MEG study of cognitive function: visual evokedmagnetic fields, motor imagery
4. Brain mechanism of Japanese kanji processing

Future Prospect
1. Cerebral localization of neuropsychological deficits(aphasia, apraxia, agnosia and amnesia): The sitesof lesions responsible for neuropsychologicaldeficits will be more precisely localized with the
MRI findings of the patient.
2. Brain functional imaging: Functional MRI study
will be advanced with development of imaging
techniques such as arterial spin labeling other than
BOLD techniques and with refinement of experimental paradigm.

Research Grants
1. JSPS Research for the Future Program (Higher Brain
Functions) [FY1997~FY 2001] ($1,095,000/¥131,000,000) "Neuroimaging contributions to theunderstanding of brain language mechanism"
2. JSPS Scientific Research (B) [FY1997~FY 1998]($79,170/ ¥9,500,000) "Neuroimaging contributions
to the understanding music"
3. JSPS Exploratory Research [FY1997] ($11,700/¥1,400,000) "Study on brain mechanism of spoken
word discrimination by MEG"
4. MEXT COE Research (Neuroscience of Music)[FY1997-2001] ($0/ ¥0) "Paradigm creation in behavioral neurology"
5. MEXT Scientific Research in Priority Areas (MindDevelopment) [FY1994-1998] ($100,000/ ¥12,000,000)"Research on disorders of cognitive development byfunctional MRI"

Select Publications
1. Takayama Y, Sugishita M, Akiguchi I, Kimura J.
Isolated acalculia due to left parietal lesion. Archivesof Neurology, 51, 286-291, 1994
2. Ishiai S, Sugishita M, Watabiki S, Nakayama T,
Kotera M, Gono S. Improvement of left unilateral
spatial neglect in a line extension task. Neurology
44, 294- 298, 1994
3. Sugishita M, Hamilton CR, Sakuma I, Henmi I.
Hemispheric representation of the central retina ofcomissurotomized subjects. Neuropsychologia 32,
399-415, 1994
4. Takayama Y, Sugishita M, Kido T, Ogawa M,
Fukuyama H, Akiguchi I. Impaired stereoacuity due
to a lesion in the left pulvinar. Journal of Neurology
Neurosurgery and Psychiatry 57, 652-654, 1994
5. Takayama Y, Kido T, Sugishita M, Ogawa M,
Akiguchi I, A case of foreign accent syndrome without aphasia caused by a lesion of the left precentral
gyrus. Neurology 44, 990, 1994
6. Takayama Y, Sugishita M. Astereopsis induced byrepetitive magnetic stimulation of occipital cortex.
Journal of Neurology 241, 522-525, 1994
7. Takayama Y, Sugishita M, Hirose S, Akiguchi I,
Anosodiaphoria for dressing apraxia: contoributory
factor to dressing apraxia. Clinical Neurology and
Neurosurgery 96, 254-256, 1994
8. Yoneda K, Sekimoto S, Yumoto M, Sugishita M. The
early component of the visual evoked magnetic field.
Neuroreport 6, 797-800, 1995
9. Sugishita M, Otomo K, Yamazaki K, Shimizu H,
Yoshioka M, Shinohara A. Dichotic listening inpatients with partial section of the corpus callosum.
Brain 118, 417-427, 1995
10. Seki K, Yajima M, Sugishita M. The efficacy of kinesthetic reading treatment for pure alexia.
Neuropsychologia, 33, 595-609, 1995
11. Sodeyama N, Tamaki M, Sugishita M. Persistentpure verbal amnesia and transient aphasia after leftthalamic infarction. Journal of Neurology 242, 289294, 1995
12. Takeda K, Sugishita M. Word length error types inJapanese left-sided neglect dyslexia.Clinical
Neurology and Neurosurgery 97, 125-130, 1995
13. Nishiyama K, Momose T, Sugishita M, Sakuta M,
Positron emission tomography of reversible intellectual impairment induced by long-term anticholinergic therapy. Journal of the Neurological Sciences,132, 89-92, 1995
14. Takayama Y, Sugishita M, Fukuyama H, Akiguchi I.
Localization in spatial vision. Clinical Neurology and
Neurosurgery 97, 249-252, 1995
15. Endo K, Makishita H, Yanagisawa N, Sugishita M.
Modality specific naming and gesture disturbances.
A case with optic aphasia, bilateral tactile aphasia,
optic apraxia and tactile apraxia. Cortex 32, 3-28,
1996
16. Sugishita M, Takayama Y, Shiono T, Yoshikawa K,
Takahashi Y. Functional maguetic resonance imaging(fMRI) during mental writing with phonograms.
Neuroreport 7, 1917-1921, 1996
17. Koike A, Simizu H, Suzuki I, Ishijima B, Sugishita M.
Preserved musical abilities following right temporal
lobectomy. Journal of Neurosurgery 85, 1000-1004,
1996
18. Nishiyama K,, Sugishita M, Kurisaki H, Sakuta M.
Reversible memory disturbance and intelligence
impairment induced by long-term anticholinergic
therapy. Internal Medicine 37, 514-518, 1998
19. Matsuda O, Saito M, Sugishita M. Cognitive deficits
of mild dementia: a comparison between dementia
of the Alzheimer’s type and vascular dementia.
Psychiatry & Clinical Neurosciences 52, 87-91, 1998
20. Kobayashi M, Takayama H, Mihara B, Sugishita M.
Partial seizure with aphasic speech arrest caused bywatching a popular animated TV program. Epilepsia
40, 652-654, 1999
21. Tanaka S, Kanzaki R, Yoshibayashi M, Kamiya T,
Sugishita M. Dichotic listening in patients with situs
inversus: brain asymmetry and situs asymmetry.
Neuropsychologia 37, 869-874, 1999
22. Ogiso T, Kobayashi K, Sugishita M. The precuneus inmotor imagery: a magnetoencephalographic study.
NeuroReport 11, 1345-1349, 2000
23. Takayama H, Kobayashi M, Sugishita M, Mihara B.
Diffusion-weighted imaging demonstrates transientcytotoxic edema involving the corpus callosum in apatient with diffuse brain injury. Clinical Neurology
and Neurosurgery 102, 135-139, 2000
24. Katanoda K, Yoshikawa K, Sugishita M. Neural substrates for the recognition of newly learned faces: afunctional MRI study. Neuropsychologia 38, 16161625, 2000
25. Katanoda K, Yoshikawa K, Sugishita M. A functional
MRI study on the neural substrates for writing.
Human Brain Mapping 13, 34-42, 2001
26. Sugishita M, Koike A, Shimizu H, Suzuki I, Ishijima
B: Right temporal lobe function. Journal of
Neurosurgery 95, 372, 2001
27. Sugishita M, Omura K. Learning Chinese characters
may improve visual recall. Perceptual and Motor
Skills 93, 579-594, 2001
28. Katanoda K, Matsuda Y, Sugishita M. A spatio-temporal regression model for the analysis of functional
MRI data. NeuroImage 16, in press, 2002

119Department of
Neuropsychiatry
Outline and Research Objectives
This department of neuropsychiatry has the longest history among all that of
Universities in Japan since 1886. In the past quarter century, unfortunately it had been in aunusual condition that two groups, “The Ward” and “The Clinic”, had made independent
clinical activities in this department and excluded each other. “The Ward” group was influenced by the movement “Anti-Psychiatry” in 1960. During this period the most part ofbudgets and personnel changes had been frozen. This unfortunate incident made the facilities of the ward and all laboratories of our department old-fashioned and obsolescent. In
1994, two groups had arrived a compromise with each other and started the care of patientsin the psychiatric ward and clinic together, however more several years were needed for theregeneration of the department. Now it has already been “normalized”, overwhelming thelong and difficult period of past quarter century and we have regenerated the productive
activities of neuropsychiatry.
Approximately 240 patients with various psychiatric disorders were admitted on lastyear (2001). Since 2002, new wards was established (34 beds in the secluded ward and 19
beds in the general ward) and this department became to be expected the more active roles
on clinical fields in this year. Occupational therapy, art therapy and group therapy are performed in the ward. For the outpatients, we have two “Day Hospital Units” in this department; Day Hospital for young patients with psychotic feature based on cognitive behavioral
approach and Day Care for autistic children using educational treatment models.
Research activities in this department are wide-ranged from social psychiatry to molecular biology. Main research activities in this department is as follows, 1) molecular biologicalstudies of psychiatric disorders, 2) neuroscientific studies of stress, 3) psychopathologlicalstudies of early schizophrenia, 4) psychophysio1ogical and neuropsychological studies ofschizophrenia and epileptic psychosis, 5) community-based psychosocial treatment ofchronic schizophrenia, 6) cognitive-developmental or pharmacological treatment of autism
and Tourette syndrome, 7) neuroimaging studies of psychiatric disorders, and 8) neuropathological studies of dementias. Starting 2000, a nation-wide Project with the Grant ofJapan Science and Technology Agency has organized under the management of thisDepartment, entitled “Molecular mechanisms underlying stress-induced brain dysfunctions
and development of diagnosis/treatment strategies on post-traumatic stress disorders(PTSD)”.

Faculties and Students
Professor and Chair
Associate Professor

Nobumasa Kato, M.D.（1998〜）
Nobuo Nakayasu, M.D.
Akira Iwanami, M.D.
Lecturer
Koichi Tsunashima, M.D.
Hitoshi Tsuda, M.D.
Rinmei Fukuda, M.D.
Associate ...............................10
Graduate Student..................13
Secretary .................................4

Past Research and Major Accomplishments
Research activity in our department ranges over
diverse fields in neuropsychiatry including biological120

and social psychiatry. Professor Kato has been conducting and supervising the researches on the effectof stress on brain using animal models. Especially,various stress-induced changes in hippocampus havebeen elucidated. Dr Nakayasu had conducted intensive psychopathological research on early symptomsof schizophrenia and proposed the notion of “incipient schizophrenia” in 1990, which could be fundamentally different from its full-blown counterpart. At
this early stage, he suggested that we could employ a
combination of specific pharmacotherapy and psychotherapy to prevent the outbreak of this devastating illness. Dr Iwanami has been utilizing event-related potentials (ERPs) to investigate the neural correlates of cognitive dysfunction in schizophrenia,
methamphetamine psychosis and other mental disor-

Grand mean ERP waveforms in patients with and without
thought disorder. (Iwanami et al. Schizophrenia Research, 2000)

Typical time courses of the NIRS variables during cognitive andphysiological tasks. (Matsuo et al. Psychological Medicine,
2002)

Future Prospects

Spatiotemporal excitation patterns of N100m sources and
SPECT imaging during musical hallucinations. (Kasai et al.
Lancet, 1999)

ders. He found that specific parameters of ERPs correlates the severity of the illness. In the field of social
psychiatry, our department was the first to introduce
Day Hospital unit in 1980s and made extensive
research on the efficacy of this treatment strategy.
Our efforts had significantly contributed to the following nationwide introduction of this treatment
approach in Japan.

Current Research
The primary activity in our department is the government-supported study in the pathogenesis ofstress-induced neuropsychiatric disorders, mainly,
post-traumatic stress disorder (PTSD). The development of novel treatment strategy will also be expected. As one of the patient population, victims of theTokyo subway sarin attack in 1995 kindly served
themselves. Especially, we employ multimodal neuroimaging techniques (electroencephalography, eventrelated potentials, magnetoencephalography, structural magnetic resonance imaging, functional magnetic
resonance imaging, magnetic resonance spectroscopy
and near infrared spectroscopy) to elucidate the individual vulnerability to this condition and the neurobiological aftereffects of psychological trauma. Other
research activities include 1) molecular genetics ofpsychiatric disorders, 2) basic neuroscience, 3) clinicaland basic psychopharmacology, 4) the development of
psychosocial treatment approach in autism, and 5)
neuropathological studies of dementias.

The Last ten years were “The Decade of Brain”.
Outstanding progress has been achieved in neuroscience in terms of scientific knowledge and technicalinnovation. In addition, the human genome project isalmost done. Now we are jumping into the 21st century, “The Century of Mind”. Psychological terms shouldbe translated into those of brain, which would provide the more solid basis for psychiatry as a branch ofneuroscience. In this context, our basic and clinicalresearch projects are in progress. For holistic understanding of patients suffering psychiatric disorders
and their treatments, multi-disciplinary studies
should be necessary under the collaboration withother neuroscience research groups.
As a university hospital, clinical practice is also
important. We should be always keen to the needs ofthe public. To meet them, we opened several specialized outpatient clinics, such as PTSD or Child andAdolescent clinic, in addition to regular services. The
teams for psychiatric emergency and palliative medicine are also being formed. In addition to clinical services, the public would expect the scientific outputs
obtained through clinical studies. The system recruiting subjects or volunteers should be established
immediately.
The curriculum for medical students and the training system for residents are now changing. Especially,
the “super-rotation” system will begin in a few years.
Our training system should be modified for residents
to acquire the better clinical clerkship.
We will continue to make our best efforts to fulfill
the three major missions of the university hospital,
that is, research, clinical service and education.

Research Grants
1. 1999:Health Science Research Grants (Research on
Brain Science)¥18,000,000

121

2. 2000:Target Oriented Brain Science Promotion
Programsupported
by
the
MEXT
2000-2002¥194,493,000
3. 2001: Japan-US Cooperative Reserch Project“Cooperative Brain Reserch
2001-2002¥2,700,000
4. 2002: Health Science Research Grants (Research on
Health Service)¥36,000,000

Select Publications
Basic
1. Jinde S, Matsui A, Morinobu A, Takahashi Y,
Tsunashima K, Noda A, Yamada N, and Kato N.
Elevated neuropeptide Y and corticotropine-releasing
factor in the brain of an novel epileptic mutant rat:
Noda epileptic rat. Brain Research 833, 286-290,
1999.
2. Shirayama Y, Hashimoto K, Matsuki H, Tsunashima
K, Iyo M, Higuchi T, and Minabe Y. Increased
expression of zif268 mRNA in rat retrosplenial cortex following administration of phencyclidine. Brain
Research 839, 180-185, 1999.
3. Nagaki S, Fukumachi F, Sakamoto Y, Higuchi H,
Miki N, Ono M, Sadamatsu M, Kato N, and Osawa
M. Upregulation of brain somatostatin and neuropeptide Y following lidocaine-induced kindling in
the rat. Brain Research 852, 470-474, 2000.
4. Kohda K, Wang Y, and Yuzaki M. Mutation of glutamate receptor motif reveals its role in gating and d2
receptor channel properties. Nature Neuroscience 3,
315-322, 2000.
5. Nishiyama M, Hong K, Mikoshiba K Poo M-m, andKato K. Polarity and input specificity of synaptic
modification: critical role of Ca2+ release from internal
stores (underlined authors equally contributed).
Nature 408, 584-588, 2000.
6. Watanabe K, Ashby CR Jr, Katsumori H, and Minabe
Y. The effect of theacute administration of various
selective 5-HT receptor antagonists on focal hippocampal seizures in freely-moving rats. European
Journal of Pharmacology 398, 239-246, 2000.
7. Yasuda S, Ishida N, Higashiyama A, Morinobu S, andKato N. Characterization of audiogenic-like seizuresin naïve rats evoked by activation of AMPA andNMDA receptors in the inferior colliculus.
Experimental Neurology 164, 396-406, 2000.
8. Minabe Y, Hashimoto K, Watanabe K, and Ashby CR
Jr. Acute and repeated administration of the selective
5-HT(2A) receptor antagonist M100907 significantly
alters the activity of midbrain dopamine neurons: anin vivo slectrophysiological study. Synapse 40, 102112, 2001.
9. Imai H, Nishimura T, Sadamatsu M, Liu Y, Kabuto
M, and Kato N. Type II glucocorticoid receptors are
involved in neuronal death and astrocyte activationinduced by trimethyltin in the rat hippocampus.
Experimental Neurology 171, 22-28, 2001.

122

10. Nishimura T, Schwarzer C, Furtinger S, Imai H, Kato
N, and Sperk G. Changes in the GABAergic system
induced by trimethyltin application in the rat.
Molecular Brain Research 97, 1-6, 2001.
11. Takahashi J, Tanaka K, Morinobu S, Fujimaki K, Li
ST, Kato K, Ohkawa M, Yamawaki S, and Kato N.
Influence of restraint stress on the expression andthe serine/threonine phosphatase activity of calcineurin in the rat brain. Synapse 40, 130-6, 2001.
12. Tsutsumi S, Akaike M, Arimitsu H, Imai H, Kato N.
Circulating corticosterone alters the rate of neuropathological and behavioral changes induced bytrimethyltin in rats. Experimental Neurology 73, 8694, 2002.

Clinical (genetics)
13. Kato T, Honda M, Kuwata S, Juji T, Kunugi H, Nanko
S, Fukuda M, and Honda Y. A novel polymorphismin the promoter region of the tumor necrosis factor
alpha gene: No association with narcolepsy.
American Journal of Medical Genetics 88, 301-304,
1999.
14. Kunugi H, Hattori M, Nanko S, Fujii K, and Kato T.
Dinucleotide repeat polymorphism in the neurotrophin-3 gene and hippocampal volume in psychoses. Schizophrenia Research 37, 271-273, 1999.
15. Kunugi H, Ishida S, Kato T, Sakai T, Tatsumi M,
Hirose T, and Nanko S. No evidence for an association of polymorphism of the tryptophan hydroxylase
gene with affective disorders or attempted suicide
among Japanese patients. American Journal of
Psychiatry 156, 774-776, 1999.
16. Kunugi H, Ishida S, Kato T, Tatsumi M, Sakai T,
Hattori M, Hirose T, and Nanko S. A functional polymorphism in the promotor region of monoamineoxidase-A gene and mood disorders. Mol Psychiatry4, 393-395, 1999.
17. Sadamatsu M, Masui A, Sakai T, Kunugi H, Nanko S,
and Kato N. Familial paroxysmal kinesigenic
choreoathetosis: an electrophysiologic and genotypic
analysis. Epilepsia 40, 942-949, 1999.
18. Kato T, Kunugi H, Nanko S, and Kato N. Associationof bipolar disorder with the 5178 polymorphism inmitochondrial DNA. American Journal of Medical
Genetics 96, 182-186, 2000.
19. Narita K, Sasaki T, Akaho R, Okazaki Y, Kusumi I,
Kato T, Hashimoto O, Fukuda R, Koyama T, Matsuo
K, Okabe Y, Nanko S, Hohjoh H, and Tokunaga K.
Human leukocyte antigen and season of birth inJapanese patients with schizophrenia. American
Journal of Psychiatry 157, 1173-1175, 2000.
20. Tomita H, Nagamitsu S, Wakui K, Fukushima Y,
Yamada K, Sadamatsu M, Masui A, Konishi T,
Matsuishi Y, Aihara M, Shimizu K, Hashimoto K,
Mineta M, Matsushima M, Tsujita T, Saito M,
Tanaka H, Tsuji S, Takagi T, Nakamura Y, Nakano S,
Kato N, Nakane Y, and Niikawa N. Paroxysmal kinesigenic choreoathetosis locus map to chromosome
16p11.2-q12.1. American Journal of Human Genetics
65, 1688-1697, 2000.

21. Kato T, Kunugi H, Nanko S, and Kato N.
Mitochondrial DNA polymorphisms in bipolar disorder. Journal of Affective Disorders 62, 151-164, 2001.
22. Tatsumi M, Sasaki T, Iwanami A, Kosuga A, Tanabe
Y, Kamijima K. Season of birth in Japanese patientswith schizophrenia. Schizophrenia Research 54, 2138, 2002.

Clinical (pharmacology/neurobiology)
23. Fukuda R, Sasaki T, Kunugi H, and Nanko S. No
changes in paired viral antibody titers during thecourse of acute schizophrenia. Neuropsychobiology
40, 57-62, 1999.
24. Kato T and Kato N. Mitochondrial dysfunction in
bipolar disorder. Bipolar Disorder 2, 180-190, 2000.
25. Fukuda R. Factors affecting serum haloperidol levelassessed by longitudinal therapeutic monitoring.
Progress in Neuropsychopharmacology and BiologicalPsychiatry 24, 1299-1318, 2000.
26. Morita S, Shimoda K, Someya T, Yoshimura Y,
Kamijima K, and Kato N. Steady-state plasma levelsof nortriptyline and its hydroxylated metabolites inJapanese: the impact of CYP2D6 genotype on thehydroxylation of nortriptyline. Journal of ClinicalPsychopharmacology 20, 141-149, 2000.
27. Iwanami A, Okajima Y, Isono H, Shinoda J, Kasai K,
Hata A, Fukuda M, Nakagome K, Kamijima K. Effectsof risperidone on event-related potentials in schizophrenic patients. Pharmacopsychiatry 34, 73-79,
2001
28. Fujimaki K, Morinobu S, Takahashi J, Yamawaki S,
Kato N, Kanno M, Okuyama N, Kawakatsu S, Otani
K, Kusumi I, and Koyama T. Nucleotide sequence
analysis of the binding site on the inositol 1,4,5trisphosphate type-1 receptor in bipolar disordernegative study. Journal of Affective Disorders 65,
139-143, 2001.
Clinical (neuroimaging)
Magnetic resonance spectroscopy
29. Hamakawa H, Kato T, Shioiri T, Inubushi T, andKato N. Quantitative proton magnetic resonancespectroscopy in the bilateral frontal lobes of patientswith bipolar disorder. Psychological Medicine 29,639-644, 1999.
30. Murashita J, Kato T, Shioiri T, Inubushi T, and Kato
N. Age-dependent alteration of metabolic responseto photic stimulation in the human brain measured
by 31P MR-spectroscopy. Brain Research 818, 72-76,
1999.
31. Kato T, Inubushi T, and Kato N. Prediction of lithium response by 31P-MRS in bipolar disorder.
International Journal of Neuropsychopharmacology
3, 83-85, 2000.
32. Murashita J, Kato T, Shioiri T, Inubushi T, and Kato
N. Altered brain energy metabolism in lithium-resistant bipolar disorder detected by photic stimulated
31P-MR spectroscopy. Psychological Medicine 30,
107-115, 2000.

Near-infrared spectroscopy (NIRS)
33. Matsuo K, Kato T, Fukuda M, and Kato N. Alterationof hemoglobin oxygenation in the frontal region inelderly depressed patients measured by near-infrared
spectroscopy. Journal of Neuropsychiatry andClinical Neuroscience 12, 465-471, 2000.
34. Matsuo K, Kato N, and Kato T. Decreased cerebral
haemodynamic response to cognitive and physiological tasks in mood disorders as shown by nearinfrared spectroscopy. Psychological Medicine 32,1029-37, 2002.
Magnetic resonance imaging (MRI)
35. Kasai K, Shenton ME, Salisbury DF, Hirayasu Y, Lee
CU, Ciszewski A, Yurgelun-Todd D, Kikinis R, Jolesz
FA, McCarley RW. Progressive decrease of left superior temporal gyrus gray matter volume in firstepisode schizophrenia. American Journal of
Psychiatry, in press.
36. Gilbertson MW, Shenton ME, Ciszewski A, Kasai K,
Lasko NB, Orr SP, Pitman RK. Smaller hippocampal
volume predicts pathologic vulnerability to psychological trauma. Nature Neuroscience, in press.
37. Onitsuka T, Shenton ME, Kasai K, Nestor PG, Toner
SK, Kikinis R, Jolesz FA, McCarley RW. Fusiform
gyrus volume reduction and facial recognition inchronic schizophrenia. Archives of General
Psychiatry, in press.
38. Kubicki M, Shenton ME, Salisbury DF, Hirayasu Y,
Kasai K, Kikinis R, Jolesz FA, McCarley RW: Voxelbased morphometric analysis of gray matter in firstepisode schizophrenia. Neuroimage, in press.
39. Anderson JE, Wible CG, McCarley RW, Jakab M,
Kasai K, Shenton ME: An MRI study of temporal
lobe abnormalities and negative symptoms in chronic schizophrenia. Schizophrenia Research, in press.
40. Lee CU, Shenton ME, Salisbury DF, Kasai K,
Onitsuka T, Dickey CC, Yurgelun-Todd D, Kikinis R,
Jolesz FA, McCarley RW: Fusiform gyrus volume
reduction in first-episode schizophrenia: an MRI
study. Archives of General Psychiatry 59, 775-781,
2002.
Clinical (neurophysiology)
41. Kasai K, Yamada H, Kamio S, Nakagome K, Iwanami
A, Fukuda M, Yumoto M, Itoh K, Koshida I, Abe O,
Kato N. Neuromagnetic correlates of impaired automatic categorical perception of speech sounds inschizophrenia. Schizophrenia Research 2002, inpress.
42. Kasai K, Asada T, Yumoto M, Takeya J, Matsuda H:
Evidence for functional abnormality in the right
auditory cortex during musical hallucinations. Lancet
354: 1703-1704, 1999.
43. Kasai K, Okazawa K, Nakagome K, Hiramatsu K,
Hata A, Fukuda M, Honda M, Miyauchi M,
Matsushita M: Mismatch negativity and N2b attenuation as an indicator for dysfunction of the preattentive and controlled processing for deviance detection123

in schizophrenia: a topographic event-related potential study. Schizophrenia Research 35: 141-156, 1999.
44. Iwanami A, Okajima Y, Kuwakado D, Isono H, Kasai
K, Hata A, Nakagome K, Fukuda M, Kamijima K.
Event-related potentials and thought disorder inschizophrenia. Schizophrenia Research 42, 187-191,
2000.
45. Kasai K, Nakagome K, Iwanami A, Fukuda M.
Neuropsychiatry and the auditory selective attention. Current Opinion in Psychiatry 14, 219-225,
2001.
46. Kamio S, Nakagome K, Murakami T, Kasai K,
Iwanami A, Hiramatsu K-I, Hata A, Watanabe A,
Fukuda M, Honda M, Kato N. Impaired suppressionof processing in schizophrenic patients suggested by
ERPs obtained in a selective attention task.
Schizophrenia Research 49, 213-221, 2001.
47. Kasai K, Nakagome K, Itoh K, Koshida I, Hata A,
Iwanami A, Fukuda M, Kato N. Impaired cortical network for preattentive detection of change in speechsounds in schizophrenia: a high-resolution eventrelated potential study. American Journal of
Psychiatry 159, 546-553, 2002.
48. Kasai K, Nakagome K, Iwanami A, Fukuda M, Itoh K,
Koshida I, Kato N. No effect of gender on tonal andphonetic mismatch negativity in normal adults
assessed by a high-resolution EEG recording.
Cognitive Brain Research 13, 305-312, 2002.
49. Kasai K, Yamada H, Kamio S, Nakagome K, Iwanami
A, Fukuda M, Yumoto M, Itoh K, Koshida I, Abe O,
Kato N. Do High or low doses of anxiolytics andhypnotics affect mismatch negativity in schizophrenic subjects? An EEG and MEG study. Clinical
Neurophysiology 113, 141-150, 2002.
50. Watanabe A, Kasai K, Nagakubo S, Kamio S,
Murakami T, Hata A, Iwanami A, Nakagome K,
Fukuda M, Kato N. Verbal and procedural memory inschizophrenia with milder symptoms: implications
for psychosocial intervention. Schizophrenia
Research 53, 263-265, 2002.

124

Department of
Neurology
Outline and Research Objectives
The Department of Neurology as a division of clinical service in The University Hospital
was established in 1964 separated from the third Department of Internal Medicine. The
facilities for research in neurology were provided from the Faculty of Medicine as a divisionof clinical research in the Institute for Brain Research until March 1997. In April 1997, ourdepartment was reorganized as a Department in the Division of Neuroscience, Graduate
School of Medicine.
Activities of our department include 1. clinical service in the out-patient clinic and theward in the University Hospital, 2. undergraduate and postgraduate education of neurology
in the Faculty of Medicine and Graduate School of Medicine, and 3. research in neuroscience in the Graduate School of Medicine. Besides a weekly ward round, a weekly clinicalconference is held for discussion on cases with difficult problems. A weekly neuroimaging
conference and journal club are also held. Moreover, a bimonthly ‘’Neuroscience Club” withstimulating discussions is held where informal lectures are given by invited distinguishedguests in the field of neuroscience from outside the University.
The Research Objectives of the Department of Neurology is to elucidate molecular mechanisms of neurological diseases and development of new therapeutic strategies based on our
understanding of molecular mechanisms of neurological diseases.

Faculties and Students
Professor and Chair Shoji Tsuji, MD, PhD (2002)
Associate Professors Shin Kwak, MD, PhD (1997)
Lecturer
Susumu Kusunoki, MD, PhD(1999)
Lecturer
Yoshikazu Ugawa, MD, PhD(1997)
Associate .................................4
Graduate student ..................18

Past Research and Major Accomplishments
The research activities have been focused on elucidation of molecular mechanisms of neurodegenerative
diseases based on molecular genetics approaches. We
took a strategy for collecting as many as families withvarious hereditary neurological diseases. We have conducted a number of projects that include linkage mapping and eventual positional cloning of the causative
genes. For the diseases in which we discovered thecausative genes, analyses of molecular mechanisms ofneurodegeneration have been conducted.
1. Identification of causative genes for neurodegenerative diseases
By linkage analysis, we have identified the loci for
Machado-Joseph disease (Nature Genet. 4:300-304,
1993), hereditary progressive dystonia (Segawa disease) (Ann. Neurol. 37:405-408, 1995) and autosomal

recessive Parkinsonism (AR-JP) (Am. J. Hum. Genet.
60:588-596, 1997). Furthermore we have identified
the causative gene for dentatorubral-pallidoluysian
atrophy (DRPLA) (Nature Genet. 6:9-13, 1994), hereditary progressive dystonia (Nature Genet. 8:236-242,
1994), spinocerebellar ataxia type 2 (SCA2), earlyonset ataxia with ocular motor apraxia and hypoalbuminemia (EAOH) (Nature Genet. 29: 184-188, 2001),
Charcot-Marie-Tooth disease type 2 (CMT2) (Cell
105:587-597, 2001) and SCA17 (Hum. Mol. Genet.
8:2047-2053, 1999, Hum. Mol. Genet. 10: 1441-1448,
2001).
2. Elucidation of molecular pathogenesis of neurodegenerative diseases
Based on identification of causative genes for theabovementioned neurodegenerative diseases, our laboratory further expanded our studies to elucidate themolecular mechanisms of neurodegeneration in these
diseases. Our major effort has been focused on molecular mechanisms of neurodegeneration in polyglutamine diseases caused by expanded CAG repeats coding for polyglutamine stretches. We have createdtransgenic mice having a full-length human mutant
DRPLA gene as a single copy, which carry various
lengths of polyglutamine stretches (Q76, Q113 andQ129 mice at the same insertion site). The expression
levels of the transgene were comparable to those ofendogenous mouse DRPLA gene. Q129 mice showed
severe neurological phenotype including ataxia,

125myoclonus and epilepsy with premature death by 16
weeks, while Q76 mice did not show any obvious
phenotypes. Q113 mice showed milder phenotypes
compared with those of Q129 mice. Given the factthat the insertion site of the transgene (human fulllength mutant DRPLA gene) is identical in these
Q129, Q113 and Q76 mice, these data strongly suggest that the variability in the phenotypes exclusively
depends on the length of the polyglutamine stretches.
Interestingly, neuronal loss was not evident in thebrains of these mice, suggesting that neuronal dysfunction not but neuronal death underlies the basic
pathophysiologic mechanisms of neurodegeneration.
Detailed analysis of these mice demonstrated that theearliest neuropathological change was intranuclear
accumulation of mutant proteins (diffuse nuclear
staining) with preferential involvement of cerebellar
nuclei, red nuclei, globus pallidus, subthalamic nuclei
and cerebral cortex. Similar findings were also confirmed in human autopsied brains of DRPLA patients.
These data strongly suggest that neuronal dysfunction is the basic molecular mechanisms of neurodegeneration in polyglutamine diseases.
Given the fact that nuclear dysfunction underlies
the molecular mechanisms of neuronal degenerationin polyglutamine diseases, we investigated nuclear
proteins that bind to expanded polyglutamine stretches. Employing yeast two-hybrid assays, we havefound TAF130 (TATA-binding protein associated factor) binds to expanded polyglutamine stretches. The
association was further confirmed by co-localizationof TAF130 and expanded polyglutamine stretches not
only in transient expression assays but also in human
autopsied brains of DRPLA and MJD patients. Since
TAF130 is involved in CREB-dependent transcriptional activation, association between TAF130 and
expanded polyglutamine stretches raised the possibilities that suppression of CREB-dependent transcription, which has been demonstrated to be essential forneuronal survival and plasticity, leads to neuronal126

dysfunctions. Suppression of CREB-dependent transcription by expanded polyglutamine stretches was
confirmed by a reporter assay, and, furthermore, bymonitoring endogenous cAMP responsive genes suchas c-FOS. Expression profiling of the Q129 mousebrains further demonstrated down-regulation of manycAMP responsive genes including c-FOS and EGR-1.
To develop therapeutic approaches for polyglutamine disease, we have investigated possibilities forabrogating suppression of CREB-dependent transcriptional activation. We found that increase in intracellular cAMP levels abrogate the suppression of the CREBdependent transcriptional activation. We have further
demonstrated that histone deacetylase (HDAC)
inhibitor has similar effects. Thus, stimulation of transcription by cAMP or HDAC inhibitors has potential
roles in therapeutic measures for polyglutamine diseases.

Current Research
1. Molecular pathogenesis of polyglutamine diseases
Current research is being focused fo the following
points. 1. Detailed analyses of transcriptional dysregulation in polyglutamine diseases. 2. Development oftherapeutic measures based on the molecular pathogenesis of polyglutamine diseases.
For elucidating the mechanisms of transcriptional
dysregulation, detailed expression profiling analysesof the Q129, Q113 and Q76 mice are being conducted.
Similar expression profiling of human autopsied
brains of patients with polyglutamine diseases is also
being conducted. Abnormalities in CREB-dependenttranscriptional activation in vivo are also being investigated by generating double transgenic mice for
DRPLA and Cre-LacZ (Lac-Z gene was inserted under
the promoter containing CRE.),For development of therapeutic approaches, thepossibilities of increasing intracellular cAMP andadministration of HDAC inhibitors are being investigated. To further establish a sensitive assay system, asensitive assay system for CREB-dependent transcriptional activation using cultured cells is being developed.
2. Molecular pathogenesis of neurodegenerative
diseases caused by deficiently in DNA repair
We have recently identified the causative gene,aprataxin, for an autosomal recessive spinocerebellar
ataxia (early-onset ataxia associated with ocularmotor apraxia and hypoalbuminemia). Preliminary
studies suggest that aprataxin binds to XRCC-1, a key
molecule involved in single strand DNA break repair.
The hypothesis that XRCC-1 is involved in the single
strand DNA break repair is further supported by thefact that aprataxin has a polynucleotide kinase-likedomain.
Since a number of neurodegenerative diseases
with mutations in the DNA repair systems havealready been found, our finding further strengthens
the functional role of DNA repair system in neurodegenerative diseases. Since the role of single strand
DNA repair system in neurodegenerative diseases has
not been well established, the future research will befocused on the roles of single strand DNA breakrepair in neurodegenerative diseases. The fact that
hypoalbuminemia becomes evident only in adulthood
further raises the possibility that transcription-coupled repair of highly actively transcribed genes suchas albumin gene might be involved in this disease.
This hypothesis is intriguing, since brain is the organwith highly active transcription activities.
3. Molecular pathogenesis of neurodegenerative
diseases with complex trait
To elucidate molecular pathogenesis of sporadic
neurodegenerative disease, we are organizing a consortium to establish a nation-wide collaborative network. To elucidate the molecular pathogenesis of suchneurodegenerative diseases with complex trait, weneed to establish large databases and to collect as
many as samples. We are focusing on amyotrophic
lateral sclerosis (sporadic ALS, and ALS in Kii
Peninsula) and multiple system atrophy. Effort isbeing made to collect as many as multiples families
for ALS and multiple system atrophy. We are performing both linkage analysis based on multiplex
families and association studies (case-control studies).
These two strategies will complement each other.

Future Prospects
For future prospects, the following projects arebeing planned.

1. Development of therapeutic measures forpolyglutamine diseases.
For accomplishing this goal, various approaches
are being planned. Considering the cascade shown in
the above Figure, the therapeutic approaches should
be focused on 1. suppression of mutant gene expressions at the upstream cascade, and 2. enhancement of
CREB-dependent transcriptional activation at thedownstream cascade will be the two major targets.
2. Molecular pathogenesis of neurodegenerative
diseases caused by deficiently in DNA repair
Detailed reconstruction studies will be conductedto elucidate the physiological functions of aprataxin.
Knock-out mice for aprataxin gene are being generated, which should be essential not only for elucidationof molecular pathogenesis, but also development oftherapeutic approached.
3. Molecular pathogenesis of neurodegenerative
diseases with complex trait
To elucidate the molecular mechanisms of neurodegeneration of sporadic diseases, the following
strategies are being planned. 1. large-scale collectionof resources including database on clinical information and genomic DNAs based on a nation-wide consortium will be essential for future research. The collection of resources will be focused on a large scalecase and controls, as well as, intensive collection ofmultiplex families. For example, we have recently
identified 4 multiplex families with multiple system
atrophy (MSA), suggesting detailed genome-wide
analyses of these families may provide important
findings as to the genetic component involved inthese diseases. These two approaches (large scale
case-control studies and intensive analyses of multiplex families) should complement, and allow us to
identify the genes involved in the pathogenesis.

Research Grants
Grant for the Research for the Future Programfrom the Japan Society for the Promotion of Science, agrant from the Research Committee for Ataxic
Diseases, the Ministry of Health, Labor and Welfare,“Identification of Genes Involved in Brain
Diseases”
FY1996
94,998FY199775,682
FY199878,658FY199985,516
FY200076,286
FY200150,000FY200247,000in thousands of yen

127Grant for Scientific Research on Priority Areas (C) Advanced Brain Science Project- from the Ministry of
Education, Culture, Sports, and Science and Technology,
Japan “Molecular mechanisms of neurodegeneration”
FY2001
66,600FY2002/10/29 59,200
in thousands of yen
Grant for Scientific Research (A) from the Ministry of
Education, Culture, Sports, and Science and Technology,
Japan
FY2002
19,200FY2003
11,300in thousands of yen
Grant from the Ministry of Health, Labor and Welfare,Japan “Study for development of therapeutic measuresfor polyglutamine diseases!
FY199824,000
FY199924,000FY200016,000in thousands of yen

Select Publications
1

2

3

4

5

6

Naruse, S., Igarashi, S., Kobayashi, H., Aoki, K.,
Inuzuka, T., Kaneko, K., Shimizu, T., Iihara, K.,
Kojima, T., Miyatake, T. and Tsuj, S.: Mis-sense
mutaiton Val→Ile in exon 17 of amyloid precursor
protein gene in Japanese familial Alzheimers disease.
Lancet 337:978-979, 1991
Wakamatsu, N., Kobayashi, H., Miyatake, T. andTsuji, S.: A novel exon mutation in the human bhexosaminidase b subunit gene affects 3 splice site
selection. J. Biol. Chem. 267:2406-2413, 1992
Tsuji, S., Kobayashi, H., Uchida, Y., Ihara, Y. and
Miyatake, T.: Molecular cloning of human growth
inhibitory factor cDNA and its down-regulation in
Alzheimers disease. EMBO J. 11:4843-4850, 1992
Igarashi, S., Tanno, Y., Onodera, O., Yamazaki, M.,
Sato, S., Ishikawa, A., Miyatani, N., Nagashima, M.,
Ishikawa, Y., Sahashi, K., Ibi, T., Miyatake, T. andTsuji, S.: Strong correlation between the number of
CAG repeats in androgen receptor genes and the clinical onset of features of spinal and bulbar muscularatrophy. Neurol. 42:2300-2302, 1992
Takiyama, Y., Nishizawa, M., Tanaka, H.,
Kawashima, S., Sakamoto, H., Karube, Y., Shimazaki,
H., Soutome, M., Endo, K., Ohta, S., Kagawa, Y.,
Kanazawa, I., Mizuno, Y., Yoshida, M., Yuasa, T.,
Horikawa, Y., Oyanagi, K., Nagai, H., Kondo, T.,
Inuzuka, T., Onodera, O. and Tsuji, S.: The gene for
Machado-Joseph disease maps to human chromosome 14q. Nature Genet. 4:300-304, 1993
Koide, R., Ikeuchi, T., Onodera, O., Tanaka, H.,
Igarashi, S., Endo, K., Takahashi, H., Kondo, R.,
Ishikawa, A., Hayashi, T., Saito, M., Tomoda, A.,
Miike, T., Naito, H., Ikuta, F. and Tsuji, S.: Unstable
expansion of CAG repeat in hereditary dentatorubral-pallidoluysian atrophy (DRPLA). Nature
Genet. 6:9-13, 1994

128

7

Okuizumi, K., Onodera, O., Tanaka, H., Kobayashi,
H., Tsuji, S., Takahashi, H., Oyanagi, K., Seki, K.,
Tanaka, M., Naruse, S., Miyatake, T., Mizusawa, H.
and Kanazawa, I.: ApoE-ε 4 and early-onset
Alzheimers. Nature Genet. 7:10-11, 1994
8 Naruse, S., Igarashi, S., Furuya, T., Kobayashi, H.,
Miyatake, T. and Tsuji, S.: Structures of the humanand mouse growth inhibitory factor-encoding genes.
Gene 144:283-287, 1994
9 Ichinose, H., Ohye, T., Takahashi, E., Seki, N., Hori,
T., Segawa, M., Nomura, Y., Endo, K., Tanaka, H.,
Tsuji, S., Fujita, K. and Nagatsu, T.: Hereditary progressive dystonia with marked diurnal fluctuation
caused by mutations in the GTP cyclohydrolase I
gene. Nature Genet. 8:236-242, 1994
10 Burke, J. R., Ikeuchi, T., Koide, R. and Tsuji, S.:
Dentatorubral-pallidoluysian atrophy and Haw River
syndrome. Lancet 344:1711-1712, 1994
11 Ikeuchi, T., Koide, R., Tanaka, H., Onodera, O.,
Igarashi, S., Takahashi, H., Kondo, R., Ishikawa, A.,
Tomoda, A., Miike, T., Sato, K., Ihara, Y., Hayabara,T,. Isa, F., Tanabe, H., Tokiguchi, S., Hayashi, M.,
Shimizu, N., Ikuta, F., Naito, H. and Tsuji, S.:
Dentatorubral-pallidoluysian atrophy closely related
to unstable expansions of trinucleotide (CAG) repeat.
Ann. Neurol. 37:769-775, 1995
12 Kaneko, K., Miyatake, T. and Tsuji, S.: Rapid andreliable diagnosis of tuberculous meningitis by PCR.
Neurol. 45:1425-1426, 1995
13 Sanpei, K., Igarashi, S., Eguchi, I., Takiyama, Y.,
Tanaka, H. and Tsuji, S.: Direct detection of expanded (CAG/CTG) repeats in the myotonin-protein
kinase genes of myotonic dystrophy patients using ahigh-stringency hybridization method. Biochem.
Biophys. Res. Commun. 212:341-346, 1995
14 Okuizumi, K., Onodera, O., Namba, Y., Ikeda, K.,
Yamamoto, T., Seki, K., Ueki, A., Nanko, S., Tanaka,
H., Takahashi, H., Oyanagi, K., Mizusawa, H.,
Kanazawa, I. and Tsuji, S.: Genetic association of thevery low density lipoprotein (VLDL) receptor genewith sporadic Alzheimers disease. Nature Genet.
11:207-209, 1995
15 Onodera, O., Oyake, M., Takano, H., Ikeuchi, T.,
Igarashi, S. and Tsuji, S.: Molecular cloning of a fulllength cDNA for dentatorubral-pallidoluysian atrophy and regional expressions of the expanded allelesin the CNS. Am. J. Hum. Genet. 57:1050-1060, 1995
16 Takiyama, Y., Igarashi, S., Rogaeva, E. A., Endo, K.,
Rogaev, E. I., Tanaka, H., Sherrington, R., Sanpei, K.,
Liang, Y., Saito, M., Tsuda, T., Takano, H., Ikeda, M.,
Lin, C., Chi, H., Kennedy, J. L., Lang, A. E., Wherrett,J. R., Segawa, M., Nomura, Y., Yuasa, T.,
Weissenbach, J., Yoshida, M., Nishizawa, M., Kidd,
K. K., Tsuji, S. and St. George-Hyslop, P. H.: Evidence
for inter-generational instability in the CAG repeat in
the MJD1 gene and for conserved haplotypes atflanking markers amongst Japanese and Caucasiansubjects with Machado-Joseph disease. Hum. Mol.
Genet. 4:1137-1146, 1995

17 Ikeuchi, T., Igarashi, S., Takiyama, Y., Onodera, O.,
Oyake, M., Takano, H., Koide, R., Tanaka, H. andTsuji, S.: Non-Mendelian transmission in dentatorubral-pallidoluysian atrophy and Machado-Joseph
disease: The mutant allele is preferentially transmitted in male meiosis. Am. J. Hum. Genet. 58:730-733,
1996
18 Takano, H., Onodera, O., Takahashi, H., Igarashi, S.,
Yamada, M., Oyake, M., Ikeuchi, T., Koide, R.,
Tanaka, H., Iwabuchi, K. and Tsuji, S.: Somatic
mosaicism of expanded CAG repeats in brains ofpatients with dentatorubral-pallidoluysian atrophy:
Cellular population-dependent dynamics of mitotic
instability. Am. J. Hum. Genet. 58:1212-1222, 1996
19 Ishikawa, A. and Tsuji, S.: Clinical analysis of 17
patients in 12 Japanese families with autosomalrecessive type juvenile parkinsonism. Neurol. 47:160166, 1996
20 Shimizu, N., Yamami, T., Nakayama, M., Ikeuchi, T.,
Koide, R. and Tsuji, S.: A sporadic case of dentatorubral pallidoluysian atrophy (DRPLA) with CAG
repeat expansion but no clinical abnormalities in thefather. J. Neurol. Neurosurg. Psychiat. 61:113-114,
1996
21 Igarashi, S., Takiyama, Y., Cancel, G., Rogaeva, E.A.,
Sasaki, H., Wakisaka, A., Zhou, Y.-X., Takano, H.,
Endo, K., Sanpei, K., Oyake, M., Tanaka, H.,
Stevanin, G., Abbas, N., Durr, A., Rogaev, E.I.,
Sherrington, R., Tsuda, T., Ikeda, M., Cassa, E.,
Nishizawa, M., Benomar, A., Julien, J., Weissenbach,J., Wang, G.-X., Agid, Y., St. George-Hyslop, P.H.,
Brice, A. and Tsuji, S.: Intergenerational instability of
the CAG repeat of the gene for Machado-Joseph disease (MJD1) is affected by the genotype of the normal chromosome: implications for the molecularmechanisms of the instability of the CAG repeat.
Hum. Mol. Genet. 5:923-932, 1996
22 Okuizumi, K., Onodera, O., Seki, K., Tanaka, H.,
Namba, Y., Ideda, K., Saunders, A. M., PericakVance, M. A., Roses, A. D. and Tsuji, S.: Lack ofassociation of very low density lipoprotein receptor
gene polymorphism with Caucasian Alzheimers disease. Ann. Neurol. 40:251-254, 1996
23 Sanpei, K., Takano, H., Igarashi. S., Sato, T., Oyake,M., Sasaki, H., Wakisaka, A., Tashiro, K., Ishida, Y.,
Ikeuchi, T., Koide, R., Saito, M., Sato, A., Tanaka, T.,
Hanyu, S., Takiyama, Y., Nishizawa, M., Shimizu, N.,
Nomura, Y., Segawa, M., Iwabuchi, K., Eguchi, I.,
Tanaka, H., Takahashi, H. and Tsuji, S.:
Identification of the spinocerebellar ataxia type 2
gene using a direct identification of repeat expansion
and cloning technique, DIRECT. Nature Genet.
14:277-284, 1996
24 Oyake, M., Onodera, O., Shiroishi, T., Takano, H.,
Takahashi, Y., Kominami, R., Moriwaki, K., Ikeuchi,T., Igarashi, S., Tanaka, H. and Tsuji, S.: Molecularcloning of murine homologue dentatorubral-pallidoluysian atrophy (DRPLA) cDNA: Strong conservation of a polymorphic CAG repeat in the murine
gene. Genomics 40:205-207, 1997

25 Matsumine, H., Saito, M., Shimoda-Matsubayashi,S., Tanaka, H., Ishikawa, A., Nakagawa-Hattori, Y.,
Yokochi, M., Kobayashi, T., Igarashi, S., Takano, H.,
Sanpei, K., Koike, R., Mori, H., Kondo, T., Mizutani,Y., Schaffer, A. A., Yamamura, Y., Nakamura, S.,
Kuzuhara, S., Tsuji, S. and Mizuno, Y.: Localizationof a gene for an autosomal recessive form of juvenile
parkinsonism to chromosome 6q25.2-27. Am. J.
Hum. Genet. 60:588-596, 1997
26 Ikeuchi, T., Asaka, T., Saito, M., Tanaka, H.,
Higuchi, S., Tanaka, K., Saida, K., Uyama, E.,
Mizusawa, H., Fukuhara, N., Nonaka, I., Takamori,
M. and Tsuji, S.: Gene locus for autosomal recessive
distal myopathy with rimmed vacuoles maps tochromosome 9. Ann. Neurol. 41:432-437, 1997
27 Takiyama, Y., Sakoe, K., Soutome, M., Namekawa,
M., Ogawa, T., Nakano, I., Igarashi, S., Oyake, M.,
Tanaka, H., Tsuji, S. and Nishizawa, M.: Single
sperm analysis of the CAG repeats in the gene for
Machado-Joseph disease (MJD1): evidence for nonMendelian transmission of the MJD1 gene and forthe effect of the intragenic CGG/GGG polymorphismon the intergenerational instability. Hum. Mol.
Genet. 6:1063-1068, 1997
28 Ikeuchi, T., Takano, H., Koide, R., Horikawa, Y.,
Honma, Y., Onishi, Y., Igarashi, S., Tanaka, H.,
Nakao, N., Sahashi, K., Tsukagoshi, H., Inoue, K.,
Takahahsi, H. and Tsuji, S.: Spinocerebellar ataxia
type 6: CAG repeat expnasion in α1A voltagedependent calcium channel gene and clinical variations in Japanese population. Ann. Neurol. 42:879884, 1997
29 Koide, R., Onodera, O., Ikeuchi, T., Kondo, R.,
Tanaka, H., Tokiguchi, S., Tomoda, A., Miike, T.,
Isa, F., Beppu, H., Shimizu, N., Watanabe, Y.,
Horikawa, Y., Shimohata, T., Hirota, K., Ishikawa, A.
and Tsuji, S.: Atrophy of the cerebellum and brainstem in dentatorubral pallidoluysian atrophy.
Influence of CAG repeat size on MRI findings.
Neurol. 49:1605-1612, 1997
30 Saito, M., Hayashi, Y., Suzuki, T., Tanaka, H.,
Hozumi, I. and Tsuji, S.: Linkage mapping of thegene for Charcot-Marie-Tooth disease type 2 to chromosome 1p (CMT2A) and the clinical features of
CMT2A. Neurol. 49:1630-1635, 1997
31 Shimohata, T., Date, H., Ishiguro, H., Suzuki, T.,
Takano, H., Tanaka, H., Tsuji, S. and Hirota, K.:
Ataxia with isolated vitamin E deficiency and retinitis pigmentosa. Ann. Neurol. 43:273, 1997
32 Igarashi, S., Koide, R., Shimohata, T., Yamada, M.,
Hayashi, Y., Takano, H., Date, H., Oyake, M., Sato,T., Sato, A., Egawa, S., Ikeuchi, T., Tanaka, H.,
Nakano, R., Tanaka, K., Hozumi, I., Inuzuka, T.,
Takahashi, H. and Tsuji, S.: Suppression of aggregate
formation and apoptosis by transglutaminase
inhibitors in cells expressing truncated DRPLA protein with an expanded polyglutamine stretch. Nature
Genet. 18:111-117, 1998
33 Onodera, O., Idezuka, J., Igarashi, S., Takiyama, Y.,
Endo, K., Takano, H., Oyake, M., Tanaka, H.,

129

Inuzuka, T., Hayashi, T., Yuasa, T., Ito, J., Miyatake,T. and Tsuji, S.: Progressive atrophy of cerebellum
and brainstem as a function of age and the size ofthe expanded CAG repeats in the MJD1 gene in
Machado-Joseph disease. Ann. Neurol. 43:288-296,
1998
34 Takano, H., Cancel, G., Ikeuchi, T., Lorenzetti, D.,
Mawad, R., Stevanin, G., Didierjean, O., Durr, A.,
Oyake, M., Shimohata, T., Sasaki, R., Koide, R.,
Igarashi, S., Hayashi, S., Takiyama, Y., Nishizawa,M., Tanaka, H., Zoghbi, H., Brice, A., Tsuji, S.: Close
associations between prevalence rates of dominantly
inherited spinocerebellar ataxias with CAG-repeat
expansions and frequencies of large normal CAG
alleles in Japanese and Caucasian populations. Am. J.
Hum. Genet. 63:1060-1066, 1998
35 Koide, T., Igarashi, S., Kikugawa, K., Nakano, R.,
Inuzuka, T., Yamada, M., Takahashi, H. and Tsuji,
S.: Formation of granular cytoplasmic aggregates incells expressing mutant Cu/Zn superoxide dismutase
associated with familial amyotrophic lateral sclerosis.
Neurosci. Lett. 257:29-32 1998
36 Okuizumi, K., Takano, H., Seki, K., Onishi, Y.,
Horikawa, Y. and Tsuji, S.: Genetic polymorphism of
the tau gene and neurodegenerative diseases with
tau pathology among Japanese. Ann. Neurol. 44:707,
1998
37 Sato, T., Oyake, M., Nakamura, K., Nakao, K.,
Fukusima, Y., Onodera, O., Igarashi, S., Takano, H.,
Kikugawa, K., lshida, Y., Shimohata, T., Koide, R.,
Ikeuchi, T., Tanaka, H., Futamura, N., Matsumura,R., Takayanagi, T., Tanaka, F., Sobue, G., Komure,O., Takahashi, M., Sano, A., Ichikawa, Y., Goto, J.,
Kanazawa, I., Katsuki, M. and Tsuji, S.: Transgenic
mice harboring a full-length human mutant DRPLA
gene exhibit age-dependent intergenerational andsomatic instabilities of CAG repeats comparable tothose in DRPLA patients. Hum. Mol. Genet. 8:99106, 1999
38 Takano, H., Koike, R., Onodera, O., Sasaki, R. andTsuji, S.: Mutational analysis and genotype-phenotype correlation of 29 unrelated Japanese patientswith X-linked adrenoleukodystrophy (ALD). Arch.
Neurol. 56:295-300, 1999
39 Ikeuchi, T., Shimohata, T., Nakano, R., Koide, R.,
Takano, H. and Tsuji, S.: A case of primary torsion
dystonia in Japan with the 3-bp (CAG) deletion in
the DYT1 gene with a unique clinical presentation.
Neurogenetics 2:189-190, 1999
40 Sato, A., Shimohata, T., Koide, R., Takano, H., Sato,
T., Oyake, M., Igarashi, S., Tanaka, K., Inuzuka, T.,
Nawa, H. and Tsuji, S.: Adenovirus-mediated expression of mutant DRPLA proteins with expanded
polyglutamine stretches in neuronally differentiated
PC12 cells. Preferential intranuclear aggregate formation and apoptosis. Hum. Mol. Genet. 8:997-1006,
1999
41 Koide, R., Kobayashi, S., Shimohata, T., Ikeuchi, T.,
Maruyama, M., Saito, M., Yamada, M., Takahashi, H.
and Tsuji, S.: A neurological disease caused by an

130

expanded CAG trinucleotide repeat in the TATAbinding protein gene: a new polyglutamine disease?
Hum. Mol. Genet. 8:2047-2053, 1999
42 Schilling, G., Wood, J. D., Duan, K., Slunt, H. H.,
Gonzales, V., Yamada, M., Cooper, J. K., Margollis, R.
L., Jenkins, N. A., Copeland, N. G., Takahashi, H.,
Tsuji, S., Price, D. L., Borchelt, D. R. and Ross, C. A.:
Nuclear accumulation of truncated atrophin-1 fragments in a transgenic mouse model of DRPLA.
Neuron 24:275-286, 1999
43 Shibasaki, Y., Tanaka, H., Iwabuchi, K., Kawasaki, S.,
Kondo H., Uekawa, K., Ueda, M., Kamiya, T.,
Katayama, Y., Nakamura, A., Takashima, H.,
Nakagawa, M., Masuda, M., Utsumi, H., Nakamuro,T., Tada, K., Kurohara, K., Inoue, K., Koike, F., Sakai,T., Tsuji, S. and Kobayashi, H.: Linkage of autosomal
recessive hereditary spastic paraplegia with mental
impairment and thin corpus callosum to chromosome 15q13-15. Ann. Neurol. 48:108-112, 2000
44 Maruyama, M., Ikeuchi, T., Saito, M., Ishikawa, A.,
Yuasa, T., Tanaka, H., Hayashi, S., Wakabayashi, K.,
Takahashi, H. and Tsuji, S: Novel mutations, pseudodominant inheritance, and possible familial affects inpatients with autosomal recessive juvenile parkinsonism. Ann. Neurol. 48:245-250, 2000
45 Shimohata, T, Nakajima, T, Yamada, M, Uchida, C,
Onodera, O, Naruse, S, Kimura, T, Koide, R, Nozaki,
K, Sano, Y, Ishiguro, H, Sakoe, K, Ooshima, T, Sato,A, Ikeuchi, T, Oyake, M, Sato, T, Aoyagi, Y, Hozumi,
I, Nagatsu, T, Takiyama, Y, Nishizawa, M, Goto, J,
Kanazawa, I, Davidson, I, Tanese, N, Takahashi, H
and Tsuji, S.: Expanded polyglutamine stretches
associated with CAG repeat diseases interact withTAFII130, interfering with CREB-dependent transcription. Nature Genet. 26:29-35, 2000
46 Zhao, C., Takita, J., Tanaka, Y., Setou, M.,
Nakagawa, T., Takeda, S., Yang, H.W., Terada, S.,
Nakata, T., Takei, Y., Saito, M., Tsuji, S., Hayashi, Y.
and Hirokawa, N.: Charcot-Marie-Tooth disease type2A caused by mutation in a microtubule motor
KIF1Bβ. Cell 105:587-597, 2001
47 Nakamura, K., Jeong, S.E., Uchihara, T., Anno, M.,
Nagashima, K., Nagashima, T., Ikeda, S., Tsuji, S.
and Kanazawa, I.: SCA17, a novel autosomal dominant cerebellar ataxia caused by an expanded polyglutamine in TATA-binding protein. Hum. Mol.
Genet. 10: 1441-1448, 2001
48 Date, H., Onodera, O., Tanaka, H., Iwabuchi, K.,
Uekawa, K., Igarashi, S., Koike, R., Hiroi, T., Yuasa,
T., Awaya, Y., Sakai, T., Takahashi,T., Nagatomo,
H., Sekijima, Y., Kawachi,I., Takiyama, Y.,
Nishizawa, M., Fukuhara, N., Saito,K., Sugano, S.
and Tsuji, S.: Early-onset ataxia with ocular motor
apraxia and hypoalbuminemia is caused by mutations in a new HIT superfamily gene. Nature Genet.
29: 184-188, 2001
49 Takahashi T., Igarashi S., Kimura T., Hozumi I.,
Kawachi I., Onodera O., Takano H., Saito M. andTsuji S.: Japanese cases of familial hemiplegic
migraine with cerebellar ataxia carrying a T666M

mutation in the CACNA1A gene. J. Neurol.
Neurosurg. Psychiat. 72: 675-680, 2002
50 Shimohata, T., Sato, A., Burke, J.R.,
Strittmatter,W.J., Tsuji S., Onodera, O.: Expanded
polyglutamine stretchers form an ‘aggresome’.
Neuroscience Letters 323: 215-218, 2002

131

Department of
Neurosurgery
Outline and Research Objectives
The Department of Neurosurgery focuses its academic activities on patient care,research, and education. Services of the Neurosurgical Clinic are devoted to the care ofpatients who suffer from brain tumors, cerebrovascular disorders, head injuries, spinal diseases, malformations of the central nervous system, and functional disturbances of thebrain. The research objectives of the department are based on these neurosurgical diseases.
Clinical research has been performed by analyzing many neurosurgical cases. The attempts
to develop new neurosurgical techniques are ongoing. Since the first Gamma-knife unit inthis country was installed in 1991, many studies have been conducted which analyze theoutcome of patients treated by this modality. To solve problems encountered in a clinicalsetting, cell biology and molecular biology of ischemic brain injury and brain tumors havebeen pursued as the two major research subjects. The research on ischemic brain damage
has been carried out using rodent ischemia models. The objective of the research has been
to determine the molecular mechanism underlying delayed neuronal death in the hippocampus following ischemia, but it has recently shifted toward the research on neuronal regeneration in the hippocampus by activation of endogenous progenitor cells. The research on
brain tumors has mainly focused on the genetic analysis of tumor specimens obtained during neurosurgery. This research strategy has particularly employed the genetic analysis ofmalignant brain tumors.

Faculty and Students
Professor and Chair Takaaki Kirino, M.D., Ph.D. (1992~)
Associate Professor Akio Morita, M.D, Ph.D.
Lecturer
Nobutaka Kawahara, M.D., Ph.D.
Lecturer
Tomoki Todo, M.D., Ph.D.
Associates ...............................8
Graduate Students ..................7
Research Students...................3
Secretaries ...............................5
Research assistants .................2
Residents.................................6

Past Research and Major Accomplishments
The research performed by our department has
been divided into three categories: clinical research,basic research on ischemic brain damage, and basic
research on brain tumors.
Research of clinical materials has been encouraged
since it may be effective in improving current andfuture patient care. Most clinical publications by thisdepartment are case studies, such as the rupture of
intracranial aneurysms (16, 17, 22, 25, 34, 41), arteriovenous or venous malformations (11, 32, 46), brain
tumors (8, 15, 21), and other disorders (13). The
results of Gamma-knife radiosurgery (6, 10, 12, 14, 19,
20, 33, 43) and the theoretical or technical aspect ofneurosurgical treatment (18, 23, 38, 50) have also132

been published. Thirty-two case reports have been
published by this department in English (not listed in
the references).
Our department has been continuously undertaking basic research as well. Currently, our basic
research consists mainly of studies on ischemic brain
damage and brain tumors. In our neurosurgical laboratory, one of the features is in vivo experiment using
our microsurgical techniques on small animals.
Another advantage of our laboratory is the availabilityof various specimens that are obtained during neurosurgical operations. In addition, we are rapidly
expanding our research facility to take advantage ofstate-of-the-art techniques in cell and molecular biology. We collaborate with various laboratories of basic
science and departments of engineering in an attempt
to expand our future possibilities in clinical andresearch activities. Some of the major accomplishments in basic research in our department have beenpossible by mutual collaborations with such laboratories.
Research on ischemic brain damage
When the brain is subjected to transient ischemia,
the first detrimental event in the brain tissue is depletion of ATP and it is followed by anoxic depolarization of the neuronal membrane and a massive
increase in the extracellular glutamate level.
Glutamate is believed to be the major factor that

Figure 1: A small, but significantly higher number of mature neurons in the CA1 are seen at Day 28 (c, c’, h). When FGF-2 and EGF
are infused into the lateral ventricle of ischemic animals, CA1 pyramidal neurons degenerates extensively at Day 7 (d, d’), but the number of neurons later after ischemia markedly increases (e, e’, j). (Cited and modified from Nakatomi et al. Cell 110:429-441,2002 5)

injures the brain tissue during the early phase of cerebral ischemia. We attempted to examine the validityof this hypothesis by inducing focal cerebral ischemia
in mice deficient in the ε1 (NR2A) subunit of the
NMDA receptor (39) in collaboration with Dr.
Mishina of the Department of Molecular
Neurobiology of this Graduate School. This experiment revealed that the brain deficient in glutamate
neurotransmission is less vulnerable to ischemic
insult.
The precise mechanism underlying ischemic brain
injury that follows the initial extracellular glutamate
surge and influx of Ca2+ remained unclear. Since a very
brief ischemic insult can induce ischemic tolerance inthe hippocampus (37), we postulated that the underlying mechanism is related to events induced by neuronal stress response. We found that significant
changes in hsp70 and ubiquitin expression occur following brief transient ischemia (28, 49). The hsp70
message and protein levels increase in toleranceinduced brain tissue. Ubiquitin expression pattern isvery similar to that of hsp70. When CA1 neurons die
following ischemia, free ubiquitin is severely depleted,and at the same time, the level of conjugated ubiquitin increases. These findings led to the hypothesisthat a disturbance in proteasomal function is the initial event that leads to neuronal apoptosis. We confirmed that under culture conditions proteasomal
inhibitors induce cytochrome-c- and caspase-3-likeprotease-induced apoptosis (24). Then, we directly
measured the proteasomal activity in the hippocampal
CA1 region. This study demonstrated the selective
proteasomal dysfunction in the CA1 sector after transient cerebral ischemia (1). This finding indicates thatthe protein-degrading system plays a major role inischemic neuronal death following brief cerebral
ischemia.

The decrease in the free ubiquitin level following
brief cerebral ischemia may be attributed to protein
degradation by activation of intracellular proteases, bydirect protein denaturation, or by an increase in thelevel of nascent protein molecules as a result of disturbed protein synthesis. We have confirmed the disturbance of protein ubiquitination and, at the sametime, inhibition of proteasomal function in CA1 neurons following brief cerebral ischemia. The mechanism of this dysfunction, however, is yet to be clarified. We hypothesized that activation of calcineurin,
an intracellular protein phosphatase, is somehow
related to proteasomal dysfunction since calcineurin
inhibitor FK506 protects CA1 neurons following
ischemia (47) and overexpression of calcineurin kills
cultured glial cells and neurons (27). This hypothesis
has not been fully tested.
Following brief ischemia, most of CA1 pyramidalcells die during 3-4 days following ischemia. The area
becomes almost devoid of neurons and finally falls inclassic gliosis. However, a close observation of thisregion for several weeks revealed a small but significant increase in the number of neurons in the CA1
subfield. Dr. Nakatomi (a graduate student of ourdepartment at that time) and Dr. Nakafuku (associate
professor, Department of Neurobiology of thisGraduate School) examined the possibility that theincreased neurons are derived from endogenous neural progenitor cells (5). The result was positive. In
addition, intraventricular infusion of FGF-2 and EGF
resulted in a more than 40% increase in neuronal population in the CA1 region. The regenerated hippocampi by this procedure exhibited electrical activity
when examined using slice preparations. The animals
that had regenerated hippocampi performed better inthe Morris water-maze task than the controls. We presented evidence that endogenous neural progenitors133

can be induced in situ to replace the hippocampal
neurons lost by ischemia. We further showed thatregenerated neurons contribute to ameliorating
ischemia-induced deficits in spatial cognitive functions, thus expanding the possibility of a novel neuronal replacement therapy for stroke. The neuronalchanges in the CA1 sector are shown in Figure 1.
We have published other experimental results on
cerebral ischemia (2, 26, 29) and on the cell biology of
cerebral blood vessels (9, 31, 48).
Research on brain tumors
Basic research on brain tumors conduced by ourdepartment has been based on specimens obtained
during surgery and those donated by affiliated hospitals. A portion of these specimens were fixed in aldehyde fixatives (formalin) and embedded in paraffin.
These materials were used for cellular analyses by
MIB-1 staining and other immunostaining techniques.
The remaining specimens were stored frozen until use
for genetic analysis. We have examined many types ofbrain tumors such as meningiomas, oligodendrogliomas, and neurinomas in NF2 (neurofibromatosis type 2) patients.
Mutation of the NF2 genes was detected in 20-30%
of sporadic meningiomas, whereas loss of heterozygosity (LOH) at chromosome 22q was found at amuch higher frequency (35). To determine the correlation of merlin loss with NF2 genetic alteration, we
performed a molecular genetic analysis of 50 sporadic
meningiomas. Findings of this study strongly support
the notion that NF2 is the sole target of 22q LOH in
meningiomas and that the loss of merlin expression isalways caused by alteration of NF2 gene, according to
the classic “two hit” theory. Oligodendrogliomas frequently, but not always, show sensitivity tochemotherapy. Recent studies demonstrated thatallelic loss of chromosome 1p is highly associatedwith this chemosensitivity. To clarify the molecularmechanism of this correlation, we examined comprehensive gene expression profiles of oligodendrogliomas with and without 1pLOH along with normal brain tissue using oligonucleotide microarray(GeneChip) (4). This study showed that biological differences between genetic subsets of oligodendroglioma are indeed reflected on the gene expression
profile. We encountered a very rare case of malignant
transformation of vestibular schwannoma following
stereotactic radiosurgery. Genotyping of this tumor
showed a TP53 mutation in the recurrent tumor that
did not exist in the original tumor, suggesting thatradiosurgery induced the malignant transformation(6). We performed molecular genetic analysis on nonselected gliomas and found that molecular genetic
analysis of 1p/19q/10q/TP53 has significant diagnostic
value, especially in detecting oligodendroglial tumors.

134

In addition, 1pLOH and TP53 mutations in gliomas
may be markers of oligodendroglial and astrocytic
pathways, respectively (7). We reported a case ofendolymphatic sac tumor, a rare adenomatous tumorof the temporal bone, in a patient with von HippelLindau (VHL) disease. Sequencing and microsatellite
analysis of DNA samples indicated that VHL gene
inactivation contributed to the oncogenesis ofendolymphatic sac tumor (30).
We examined brain tumor specimens to analyze
MEN1 gene mutation (36), telomerase activity (40),
MIB-1 in central neurocytoma (42), and proliferationof chordoma (44). We also published experimental
data on cultured glioma cells (3, 45).

Current Research
We are continuously conducting clinical researches
to improve neurosurgical technologies and future
patient care in the neurological field. The department
places strong emphasis on publishing clinical reports
and particularly encourages young colleagues to do so.
The objective of current clinical research in neurosurgery is gradually shifting to the improvement ofless invasive neurosurgical procedures using microendoscope, navigation and robotic manipulators. This
is now under way in collaboration with several
departments in the Faculty of Engineering of our university. The department is involved in the nationwide epidemiological study of the incidentally-discovered unruptured cerebral aneurysms (Unruptured
Cerebral Aneurysm Study of Japan, or UCAS Japan),and functions as the central office of this study.
We have been involved in experiments on themolecular mechanism underlying delayed neuronal
death found in the hippocampal CA1 region following
ischemia. Although we believe that this is a goodmodel system for studying neuronal protection,
experimental results obtained by us and other groups
suggest that neuronal protection is extremely difficult
and we are so far unable to propose clinically applicable methods of treatment. On the other hand, itturned out to be possible to induce practically meaningful regeneration of neurons in the hippocampal
CA1 region by activating endogenous neural progenitor cells. Given these experimental results, we are
now attempting to focus our experimental resourceson experiments of regeneration of CA1 neurons.
We have been engaged in the genetic analysis ofbrain tumors. This strategy has been particularly
effective for this department since comprehensive
genetic analysis has been rapidly introduced and we
can collect sufficient amounts of specimens during
neurosurgical operations. This strategy is being continued but the focus on brain tumor experiments isnow shifting toward research on treatment of malig-nant brain tumors because Dr. Todo who has expertise in this field has just joined our department inJanuary, 2003. He plans to start a clinical trial withgenetically engineered retrovirus for the treatment ofmalignant brain tumors.

5. Research grant from SORST (Solution Oriented
Research for Science and Technology) by Japan
Science and Technology Corporation "Neuronal
regeneration in the hippocampus following delayed
neuronal death", 2002-, ¥75,000,000

Future Prospects

Selected Papers (1995-2002)

We will continue to carry out our three major
research projects: i.e., clinical research, research on
ischemic brain damage, and research on brain tumors.
However, the main focus of each project may change.
The future projects that we expect to conduct in thenext 4-5 years are:

1. Asai A, Tanahashi N, Qiu JH, Saito N, Chi S,
Kawahara N, Tanaka K, Kirino T. Selective proteasomal dysfunction in the hippocampal CA1 region after
transient forebrain ischemia. J Cereb Blood Flow
Metab 22, 705-710, 2002
2. Kawahara N, Kawai K, Toyoda T, Nakatomi H,
Furuya K, Kirino T. Cardiac arrest cerebral ischemia
model in mice failed to cause delayed neuronal deathin the hippocampus. Neurosci Lett 322, 91-94, 2002
3. Mochizuki T, Asai A, Saito N, Tanaka S, Katagiri H,
Asano T, Nakane M, Tamura A, Kuchino Y, Kitanaka
C, Kirino T. Akt protein kinase inhibits non-apoptotic programmed cell death induced by ceramide. J Biol
Chem 277, 2790-2797, 2002
4. Mukasa A, Ueki K, Matsumoto S, Tsutsumi S,
Nishikawa R, Fujimaki T, Asai A, Kirino T,
Aburatani H. Distinction in gene expression profilesof oligodendrogliomas with and without allelic lossof 1p. Oncogene 21, 3961-3968, 2002
5. Nakatomi H, Kuriu T, Okabe S, Yamamoto S, Hatano
O, Kawahara N, Tamura A, Kirino T, Nakafuku M.
Regeneration of hippocampal pyramidal neurons
after ischemic brain injury by recruitment of endogenous neural progenitors. Cell 110, 429, 2002
6. Shin M, Ueki K, Kurita H, Kirino T. Malignant transformation of a vestibular schwannoma after gamma
knife radiosurgery. Lancet 360, 309-310, 2002
7. Ueki K, Nishikawa R, Nakazato Y, Hirose T, Hirato J,
Funada N, Fujimaki T, Hojo S, Kubo O, Ide T, Usui
M, Ochiai C, Ito S, Takahashi H, Mukasa A, Asai A,
Kirino T. Correlation of histology and molecular
genetic analysis of 1p, 19q, 10q, TP53, EGFR, CDK4,
and CDKN2A in 91 astrocytic and oligodendroglial
tumors. Clin Cancer Res 8, 196-201, 2002
8. Asano S, Ueki K, Suzuki I, Kirino T. Clinical featuresand medical treatment of male prolactinomas. Acta
Neurochir (Wien) 143, 465-470, 2001
9. Hoya K, Asai A, Sasaki T, Kimura K, Kirino T.
Expression of smooth muscle proteins in cavernousand arteriovenous malformations. Acta Neuropathol(Berl) 102, 257-263, 2001
10. Kawai K, Suzuki I, Kurita H, Shin M, Arai N, Kirino
T. Failure of low-dose radiosurgery to control temporal lobe epilepsy. J Neurosurg 95, 883-887, 2001
11. Kurita H, Shin M, Ueki K, Kawamoto S, Kirino T.
Congestive brain oedema associated with a pial arteriovenous malformation with impaired venous
drainage. Acta Neurochir (Wien) 143, 339-342, 2001
12. Kurita H, Kawahara N, Asai A, Ueki K, Shin M,
Kirino T. Radiation-induced apoptosis of oligodendrocytes in the adult rat brain. Neurol Res 23, 869874, 2001

1. Clinical research
The aim will be the technical improvement of lessinvasive neurosurgical procedures using micro-endoscope, navigation, robotic manipulators as well asother devices. We will continue large-scale clinicalstudies such as UCAS Japan. Clinical studies on neurosurgical cases will be continuously encouraged.
2. Research on ischemic brain damage:The main objective will be regeneration of hippocampal neurons following brief cerebral ischemia.
We will focus on the feasibility of activation of endogenous neural progenitors under a clinical setting. The
research on the mechanism underlying ischemic brain
damage and ischemic tolerance will be continued.
3. Research on brain tumors:
We will continue genetic analysis of brain tumor
specimens. Along with this investigation, we will
start clinical research on the treatment of malignant
brain tumors.

Research Grants
1. Research grant from CREST (Core Research for
Evolutional Science and Technology) by Japan
Science and Technology Corporation"Molecular mechanism of delayed neuronal death",
1998-2002, ¥265,201,000
2. Research grant of 21st century medical development
and promotion by the Ministry of Health, Labor andWelfare "Follow-up study of unruptured cerebral
aneurysms diagnosed by brain check-up clinics",
1999-2001, ¥24,000,000
3. Research Grants in Natural Sciences by the
Mitsubishi Foundation "Study of ischemic neuronal
death and ischemic tolerance based on functional
genomic/proteomic analysis", 2001, ¥7,000,000
4. Grant-in-Aid for Scientific Research (A) "Mechanismof ischemic cerebral damage by systematic expression analysis of genome and proteome", 2001-,¥30,000,000

13513. Oya S, Tsutsumi K, Ueki K, Kirino T. Reinduction ofhyponatremia to treat central pontine myelinolysis.
Neurology 57, 1931-1932, 2001
14. Shin M, Kurita H, Sasaki T, Kawamoto S, Tago M,
Kawahara N, Morita A, Ueki K, Kirino T. Analysis oftreatment outcome after stereotactic radiosurgery forcavernous sinus meningiomas. J Neurosurg 95, 435439, 2001
15. Sora S, Ueki K, Saito N, Kawahara N, Shitara N,
Kirino T. Incidence of von Hippel-Lindau disease inhemangioblastoma patients: the University of Tokyo
Hospital experience from 1954-1998. Acta Neurochir(Wien) 143, 893-896, 2001
16. Tsutsumi K, Ueki K, Kirino T. Recurrent subarachnoid hemorrhage. J Neurosurg 94, 541-542, 2001
17. Tsutsumi K, Ueki K, Morita A, Usui M, Kirino T. Riskof aneurysm recurrence in patients with clipped cerebral aneurysms: results of long-term follow-up
angiography. Stroke 32, 1191-1194, 2001
18. Fujimaki T, Kirino T. Combined transhorizontalsupracerebellar approach for microvascular decompression of trigeminal neuralgia. Br J Neurosurg 14,
531-534, 2000
19. Kurita H, Ueki K, Shin M, Kawamoto S, Sasaki T,
Tago M, Kirino T. Headaches in patients with radiosurgically treated occipital arteriovenous malformations. J Neurosurg 93, 224-228, 2000
20. Kurita H, Kawamoto S, Sasaki T, Shin M, Tago M,
Terahara A, Ueki K, Kirino T. Results of radiosurgery
for brain stem arteriovenous malformations. J Neurol
Neurosurg Psychiatry 68, 563-570, 2000
21. Matsuno A, Katakami H, Nagashima T, Teramoto A,
Osamura RY, Kirino T. Growth hormone-releasing
hormone expression in pituitary somatotroph adenomas, studied by immunohistochemistry and in situ
hybridization using catalyzed signal amplification
system. Hum Pathol 31, 789-794, 2000
22. Nakatomi H, Segawa H, Kurata A, Shiokawa Y,
Nagata K, Kamiyama H, Ueki K, Kirino T.
Clinicopathological study of intracranial fusiform anddolichoectatic aneurysms : insight on the mechanismof growth. Stroke 31, 896-900, 2000
23. Nishihara T, Teraoka A, Morita A, Ueki K, Takai K,
Kirino T. A transparent sheath for endoscopic surgery and its application in surgical evacuation of
spontaneous intracerebral hematomas. Technicalnote. J Neurosurg 92, 1053-1055, 2000
24. Qiu JH, Asai A, Chi S, Saito N, Hamada H, Kirino T.
Proteasome inhibitors induce cytochrome c-caspase3-like protease-mediated apoptosis in cultured cortical neurons. J Neurosci 20, 259-265, 2000
25. Tsutsumi K, Ueki K, Morita A, Kirino T. Risk of rupture from incidental cerebral aneurysms. J Neurosurg
93, 550-553, 2000
26. Zhu L, Saito N, Abe O, Okubo T, Yamada H,
Kawahara N, Asai A, Kirino T. Changes in the apparent diffusion coefficient of water and T2 relaxation
time in gerbil hippocampus after mild ischemia.
Neuroreport 11, 3333-3336, 2000

136

27. Asai A, Qiu J, Narita Y, Chi S, Saito N, Shinoura N,
Hamada H, Kuchino Y, Kirino T. High level calcineurin activity predisposes neuronal cells to apoptosis. J Biol Chem 274, 34450-34458, 1999
28. Ide T, Takada K, Qiu JH, Saito N, Kawahara N, Asai
A, Kirino T. Ubiquitin stress response in postischemic hippocampal neurons under nontolerant andtolerant conditions. J Cereb Blood Flow Metab 19,
750-756, 1999
29. Kawahara N, Mishima K, Higashiyama S, Taniguchi
N, Tamura A, Kirino T. The gene for heparin-binding
epidermal growth factor-like growth factor is stressinducible: its role in cerebral ischemia. J Cereb Blood
Flow Metab 19, 307-320, 1999
30. Kawahara N, Kume H, Ueki K, Mishima K, Sasaki T,
Kirino T. VHL gene inactivation in an endolymphatic
sac tumor associated with von Hippel-Lindau disease. Neurology 53, 208-210, 1999
31. Kim P, Yoshimoto Y, Nakaguchi H, Mori T, Asai A,
Sasaki T, Kirino T, Nonomura Y. Increased sarcolemmal permeability in the cerebral artery during chronic spasm: an assessment using DNA-binding dyes
and detection of apoptosis. J Cereb Blood Flow
Metab 19, 889-897, 1999
32. Kurita H, Kirino T. A longitudinal study of patientswith venous malformations. Neurology 52, 11091110, 1999
33. Kurita H, Sasaki T, Tago M, Kaneko Y, Kirino T.
Successful radiosurgical treatment of arteriovenous
malformation accompanied by venous malformation.
AJNR Am J Neuroradiol 20, 482-485, 1999
34. Tsutsumi K, Ueki K, Usui M, Kwak S, Kirino T. Riskof subarachnoid hemorrhage after surgical treatmentof unruptured cerebral aneurysms. Stroke 30, 11811184, 1999
35. Ueki K, Wen Bin C, Narita Y, Asai A, Kirino T. Tight
association of loss of merlin expression with loss ofheterozygosity at chromosome 22q in sporadic
meningiomas. Cancer Res 59, 5995-5998, 1999
36. Wenbin C, Asai A, Teramoto A, Sanno N, Kirino T.
Mutations of the MEN1 tumor suppressor gene insporadic pituitary tumors. Cancer Lett 142, 43-47,
1999
37. Kawahara N, Ide T, Saito N, Kawai K, Kirino T.
Propentofylline potentiates induced ischemic tolerance in gerbil hippocampal neurons via adenosine
receptor. J Cereb Blood Flow Metab 18, 472-475,
1998
38. Kim P, Wakai S, Matsuo S, Moriyama T, Kirino T.
Bisegmental cervical interbody fusion using hydroxyapatite implants: surgical results and long-term
observation in 70 cases. J Neurosurg 88, 21-27, 1998
39. Morikawa E, Mori H, Kiyama Y, Mishina M, Asano
T, Kirino T. Attenuation of focal ischemic brain
injury in mice deficient in the epsilon1 (NR2A) subunit of NMDA receptor. J Neurosci 18, 9727-9732,
1998
40. Sano T, Asai A, Mishima K, Fujimaki T, Kirino T.
Telomerase activity in 144 brain tumours. Br J
Cancer 77, 1633-1637, 1998

41. Tsutsumi K, Ueki K, Usui M, Kwak S, Kirino T. Riskof recurrent subarachnoid hemorrhage after complete obliteration of cerebral aneurysms. Stroke 29,
2511-2513, 1998
42. Fujimaki T, Matsuno A, Sasaki T, Toyoda T,
Matsuura R, Ogai M, Kitanaka C, Asai A, Matsutani
M, Kirino T. Proliferative activity of central neurocytoma: measurement of tumor volume doubling time,
MIB-1 staining index and bromodeoxyuridine labeling index. J Neurooncol 32, 103-109, 1997
43. Kurita H, Sasaki T, Kawamoto S, Taniguchi M,
Terahara A, Tago M, Kirino T. Role of radiosurgery
in the management of cavernous sinus meningiomas.
Acta Neurol Scand 96, 297-304, 1997
44. Matsuno A, Sasaki T, Nagashima T, Matsuura R,
Tanaka H, Hirakawa M, Murakami M, Kirino T.
Immunohistochemical examination of proliferative
potentials and the expression of cell cycle-related
proteins of intracranial chordomas. Hum Pathol 28,714-719, 1997
45. Mishima K, Asai A, Kadomatsu K, Ino Y, Nomura K,
Narita Y, Muramatsu T, Kirino T. Increased expression of midkine during the progression of humanastrocytomas. Neurosci Lett 233, 29-32, 1997
46. Furuya K, Sasaki T, Suzuki I, Kim P, Saito N, Kirino
T. Intramedullary angiographically occult vascularmalformations of the spinal cord. Neurosurgery 39,1123-1130, 1996
47. Ide T, Morikawa E, Kirino T. An immunosuppressant, FK506, protects hippocampal neurons from
forebrain ischemia in the mongolian gerbil. Neurosci
Lett 204, 157-160, 1996
48. Kim P, Yoshimoto Y, Iino M, Tomio S, Kirino T,
Nonomura Y. Impaired calcium regulation of smoothmuscle during chronic vasospasm following subarachnoid hemorrhage. J Cereb Blood Flow Metab16, 334-341, 1996
49. Morimoto T, Ide T, Ihara Y, Tamura A, Kirino T.
Transient ischemia depletes free ubiquitin in the gerbil hippocampal CA1 neurons. Am J Pathol 148, 249257, 1996
50. Chang HS, Kirino T. Quantification of operative benefit for unruptured cerebral aneurysms: a theoretical
approach. J Neurosurg 83, 413-420, 1995

137

Department of MolecularPreventive Medicine
Outlines and Research objectives
This department originates from The Department of Hygiene which was first established by Prof. Masanori Ogata in 1885. Prof. Ogata is the first Japanese bacteriologist who
was trained by Prof. Pettenkofer in Munich. The Department of Microbiology was separated
in 1920. I (K.M.) am the 8th professor of this department.
Main current research interests are as follows:
1. Molecular pathogenesis of inflammatory and immune diseases focusing on chemokinesand dendritic cells, and development of novel therapeutics for inflammatory andimmune diseases.
2. Molecular mechanism of leukocyte trafficking in vivo and chemotaxis in vitro.
3. Development of novel, effective ways of dendritic cells(DC)-based vaccination for infectious and inflammatory diseases as well as cancer.
4. Identification of novel targets for intervention therapy and prevention of inflammatory
and immune diseases through completing comprehensive gene expression profiling of
human leukocyte subsets.

Faculties and Students
Professor and Chair

Kouji Matsushima MD and PhD(1996 appointed)
Associated Professor Sho Ishikawa MD and PhD
Associate .................................2
Assistant .................................1
Postdoctoral Fellows...............2 (JSP) 1 (SORST, JST)
Graduate Students ..................9
Graduate Students from outside ....8
Visiting Scientists ...................5
Technical Assistants ..........1+1 (SORST, JST)
Secretary .................................1 (SORST, JST)

Past Major Accomplishments
Chemokine, chemotactic cytokine family now consists of over 40 and is subdivided into four subfamilies
based on the location of the very conserved first two cysteine residues. Interleukin 8 (IL 8), which was purified
based on in vitro neutrophil chemotactic activity andmolecularly cloned by me in collaboration with Teizo
Yoshimura in 1987, is a prototype of CXC chemokines.
On the other hand, MCAF/MCP-1, which was purified
based on monocyte chemotactic activity and molecularlycloned by me and Teizo Yoshimura independently in
1989, is a prototype of CC chemokines.
From the beginning of the 1990’s we and several
other groups initiated the studies to establish thepathophysiological roles of chemokines in various animal inflammation models using specific blocking antibodies against chemokines. We reported the essential

138

involvement of IL 8 in recruiting neutrophils in acuteinflammation models such as LPS/IL-1 induced dermatitis, immune-complex induced acute glomerulonephritis, lung reperfusion injury, acute respiratory
distress syndrome and brain infarct; and that intervention of IL 8 leads to the prevention of neutrophil infiltration-associated tissue injury. We also revealed thepivotal role of MCAF/MCP-1 in recruiting monocytes/macrophages in chronic inflammatory diseases
through working on chronic glomerulonephritis
caused by anti-glomerular basement membrane antibody, thickening of endothelium after carotid arteryinjury as an athelosclerosis model, and monocrotalineinduced pulmonary hypertension model in rats. Later,
the pivotal role of IL 8 and MCAF/MCP-1 in recruiting
neutrophils and monocytes, respectively was essentially confirmed by other groups analyzing gene targeted
mice for IL 8 receptor homologue, and JE (murine
homologue of MCAF/MCP-1) and its receptor CCR2.
Recent discovery of numerous novel chemokines
by others through signal sequence trap method and
EST data base, most of which are chemoattractantsfor immune cells such as subclasses of T lymphocytes,
B lymphocytes and DC, has changed our understanding of the role of chemokines in host defense responses. For example, CD4+CD45RA+naive T lymphocytes
express CCR7. CD4+CD45RO+memory T lymphocytes express CXCR3 and CCR5 on Th1 subset andCCR4 on Th2 subset. We generated a monoclonal
antibody against human CCR4 and established thatabout 20% of CD4+ memory T lymphocytes in the circulation of normal individuals express CCR4 andFigure 1: Peripheral and regional DC-Th cell clusters as the sites of Th1 cell generation regulatedby DC-derived chemokinesthese cells are committed Th2 population producing
preferentially Th2 cytokines without any cultures topolarize into Th2 cells in vitro and the percentage ofthis population is much increased in atopic diseases
such as atopic dermatitis and asthma. We also reported a pivotal role of the ligands for CCR4, TARC and
MDC in causing and regulating atopic diseases inmurine models.
Molecular pathogenesis of inflammatory andimmune diseases
Chemokines in a murine model of bacteria-induced
fulminant heapatitis
Propionibacterium acnes (P.acnes) is the most probable causing bacterium of sarcoidosis. In this model,
administration of P.acnes causes numerous granuloma
formation in the sinusoidal area of the liver, and subsequent challenge of the mice with low dose of LPS
induces massive hepatic injury around granuloma. We
first revealed that Th1 type immune response dominates in granuloma formation in terms of
chemokine/chemokine receptor expression as well as
cytokine production, whereas dramatic shift to Th2
response occurs immediately after LPS administration.
Administration of the antibody against TARC (a ligandof CCR4, Th2 chemokine receptor) significantly inhibited liver injury. This is the first presentation that the
rapid local shift from Th1 to Th2 by chemokines leads
to severe tissue injury. We next identified TARC producing cells at the granuloma sites to be DC. We, thenextensively analyzed the mechanism of granuloma formation, and revealed that numerous CCR1/5+DC-precursors appear shortly after P.acnes administration in
the circulation and rapidly migrate into sub-sinusoidal
area (Disse’s space) in response to MIP1a produced by
Kupffer cells to participate in the initial granuloma formation. Antigen-laden DC matured at the granuloma
sites start to express CCR7, and subsequently migrate
to portal area and the draining lymph nodes(hepatic/celiac lymph nodes) in response to SLC produced by lymphatic endothelium. We revealed that thefirst immune response occurs at the portal area andinflammation-associated lymphoid tissue appears at

the portal area which we termed “portal tract-associated lymphoid tissue, PALT. We also showed directlythat Th1 polarization occurrs at the draining lymphnodes by visualizing IFN-gamma producing CD4+T cells
clustered with DC in the paracortex of lymph nodes.
We provided evidence that IP-10 produced by migrated mature DC in the hepatic lymph nodes regulates
DC-Th1 cluster formation and controls the exit ofmemory/effector T cells from the lymph nodes tohome to the liver to complete granuloma formation(Figure 1). This is the first report describing the existence of DC at the granuloma sites and the identification of inflammation-associated DC precursors in thecirculation. This implies that even the exit/release ofmemory/effector T cells from the peripheral lymph
nodes is regulated by chemokines. This study has provided a new concept that chemokine-recruited DC
tightly link inflammation (innate immunity) andacquired immunity. Inflammation and immunity
should not be conceptually separated any more, andgranuloma formation is not a mere end-point of chronic inflammation, rather a very active immune site.
1; Initiation and induction of DC-naive T cell clusters by ELC(CCL19) or SLC(CCL21). 2; Amplificationof DC-nonpolarized T cell clusters by MDC(CCL22). 3;
Promotion and retention of DC-Th1 cell clusters (thisstudy). These clusters may develop in the given
spaces of the paracortex called “cytokine fields”. DCderived chemokines may regulate T cell traffic
depending on their state of activation between thesegiven spaces to magnify the effective immune
responses. After leaving the LNs, Th1 cells migrate
into the liver through the circulation and 4. re-form
clusters with peripheral tissue-resided inflammatory
DCs (granuloma formation) possibly by MIP1a(CCL3)
or Mig(CXCL9). Th1 cells can further proliferate andproduce higher amount of IFN-γ at the periphery of
the granulomas to complete polarization. HEV, highendothelial venule.

Current Research
DC migration pathway so far described is limited139

to the idea that tissue immature DC such as
Langerhans cells mature after antigen capturing andstart to express CCR7 to be recruited to draining
lymph nodes through afferent lymphatics. There isneither direct presentation of DC precursor migration
into any organs nor evidence of the entry of DC into
secondary lymph nodes through HEV (high endothelial venule) directly from the circulation. We have now
identified DC precursors which rapidly appear in thecirculation in inflammation. We are currently studying the following issues:
Mechanism of rapid release of DC precursors frombone marrow---identification of chemokine(s)involved.
Heterogeneity of the identified DC precursors andfate of DC precursors after administering into normal
and inflamed mice. Most of cells are
CD11c+CD11b+myeloid type DC, but some of thecells express L-selectin and B220. Therefore, some ofthese cells may directly enter peripheral lymph nodes
through HEV to act as tolerance-inducing DC.
Identification of homing molecules to the lymph
nodes.
Examination of the possible usefulness of theidentified DC precursors for vaccination and antigenspecific tolerance induction.
Future Prospective: Investigation of “lymph nodehoming” circulating DC precursors may provide a clue
to the mechanism of systemic tolerance induction andan approach to establish effective way of DC-based
vaccination to cancer and microbes. We will further
examine the role of chemokines in Th2 polarization,
CTL generation and the recruitment and maintainance of memory state of CD4/8 T lymphocytes.
We will also establish the critical role of TARC/MDC
to recruit CCR4+CD4+CD25+ so called regulatory T
cells in cancer and chronic infection models. The clinical development of humanaized monoclonal antibody
against human CCR4 will be tried in a collaboration
with a pharmaceutical. This antibody is expected toeliminate by ADCC CCR4+cells such as Th2, regulatory T cells and adult leukemia cells in a few hours invivo. Therefore, this antibody treatment may be useful for the treatment of various atopic diseases. This
antibody may also be useful to recover patients from
immune-suppression to respond more to DC-based
vaccination. This antibody may also become an effective drug for curing ATL patients.

Research Grants
Research Grants from JSP
1996
27,600,000 JPY
1997
11,100,000 JPY
2001
7,000,000 JPY

140

2002
20,300,000 JPY
Core Research of Evolutional Science and Technology(CREST, JST)
1996-2001 Total 600,000,000 JPY (about 5 million USD)
Solution Oriented Research for Science and Technology(SORST, JST)
15,000,000 JPY (about 120,000 USD)
70,000,000 JPY (about 500,000 USD)

50 Select Publications
1. Kimura H, Kasahara Y, Kurosu K, Sugito K, Takiguchi
Y, Terai M, Mikata A, Natsume M, Mukaida N,
Matsushima K, Kuriyama T. Alleviation of monocrotaline-induced pulmonary hypertension by antibodies to monocyte chemotactic and activating
factor/monocyte chemoattractant protein-1. Lab
Invest. 1998;78:571-581
2. Kuno K, Matsushima K. ADAMTS-1 protein anchorsat the extracellular matrix through the thrombospondin type I motifs and its spacing region. J Biol
Chem. 1998;273:13912-13917
3. Sonoda Y, Mukaida N, Wang JB, Shimada-Hiratsuka
M, Naito M, Kasahara T, Harada A, Inoue M,
Matsushima K. Physiologic regulation of postovulatory neutrophil migration into vagina in mice by a C-XC chemokine(s). J Immunol. 1998;160:6159-6165
4. Tachibana K, Hirota S, Iizasa H, Yoshida H, Kawabata
K, Kataoka Y, Kitamura Y, Matsushima K, Yoshida N,
Nishikawa S, Kishimoto T, Nagasawa T. The
chemokine receptor CXCR4 is essential for vascularization of the gastrointestinal tract. Nature.
1998;393:591-594
5. Wang J, Harada A, Matsushita S, Matsumi S, Zhang Y,
Shioda T, Nagai Y, Matsushima K. IL-4 and a glucocorticoid up-regulate CXCR4 expression on humanCD4+ T lymphocytes and enhance HIV-1 replication.
J Leukoc Biol. 1998;64:642-649
6. Yoneyama H, Harada A, Imai T, Baba M, Yoshie O,
Zhang Y, Higashi H, Murai M, Asakura H,
Matsushima K. Pivotal role of TARC, a CC
chemokine, in bacteria-induced fulminant hepatic
failure in mice. J Clin Invest. 1998;102:1933-1941
7. Zhang Y, Harada A, Wang JB, Zhang YY, Hashimoto S,
Naito M, Matsushima K. Bifurcated dendritic cell differentiation in vitro from murine lineage phenotypenegative c-kit+ bone marrow hematopoietic progenitor cells. Blood. 1998;92:118-128
8. Hashimoto S, Suzuki T, Dong HY, Nagai S, Yamazaki
N, Matsushima K. Serial analysis of gene expression
in human monocyte-derived dendritic cells. Blood.
1999;94:845-852
9. Hashimoto S, Suzuki T, Dong HY, Yamazaki N,
Matsushima K. Serial analysis of gene expression inhuman monocytes and macrophages. Blood.
1999;94:837-844
10. Imai T, Nagira M, Takagi S, Kakizaki M, Nishimura
M, Wang J, Gray PW, Matsushima K, Yoshie O.
Selective recruitment of CCR4-bearing Th2 cells
toward antigen-presenting cells by the CC

chemokines thymus and activation-regulated
chemokine and macrophage-derived chemokine. Int
Immunol. 1999;11:81-88
11. Kuno K, Terashima Y, Matsushima K. ADAMTS-1 isan active metalloproteinase associated with theextracellular matrix. J Biol Chem. 1999;274:1882118826
12. Murai M, Yoneyama H, Harada A, Yi Z, Vestergaard
C, Guo B, Suzuki K, Asakura H, Matsushima K.
Active participation of CCR5(+)CD8(+) T lymphocytes in the pathogenesis of liver injury in graft-versus-host disease. J Clin Invest. 1999;104:49-57
13. Ogata M, Zhang Y, Wang Y, Itakura M, Zhang YY,
Harada A, Hashimoto S, Matsushima K. Chemotactic
response toward chemokines and its regulation bytransforming growth factor-beta1 of murine bonemarrow hematopoietic progenitor cell-derived different subset of dendritic cells. Blood. 1999;93:32253232
14. Tsuji H, Mukaida N, Harada A, Kaneko S, Matsushita
E, Nakanuma Y, Tsutsui H, Okamura H, Nakanishi
K, Tagawa Y, Iwakura Y, Kobayashi K, Matsushima
K. Alleviation of lipopolysaccharide-induced acuteliver injury in Propionibacterium acnes-primed IFNgamma-deficient mice by a concomitant reduction of
TNF-alpha, IL-12, and IL-18 production. J Immunol.
1999;162:1049-1055
15. Vestergaard C, Yoneyama H, Murai M, Nakamura K,
Tamaki K, Terashima Y, Imai T, Yoshie O, Irimura T,
Mizutani H, Matsushima K. Overproduction of Th2specific chemokines in NC/Nga mice exhibiting
atopic dermatitis-like lesions. J Clin Invest.
1999;104:1097-1105
16. Zhang Y, Zhang YY, Ogata M, Chen P, Harada A,
Hashimoto S, Matsushima K. Transforming growth
factor-beta1 polarizes murine hematopoietic progenitor cells to generate Langerhans cell-like dendritic
cells through a monocyte/macrophage differentiation
pathway. Blood. 1999;93:1208-1220
17. Hashimoto SI, Suzuki T, Nagai S, Yamashita T,
Toyoda N, Matsushima K. Identification of genes
specifically expressed in human activated andmature dendritic cells through serial analysis of gene
expression. Blood. 2000;96:2206-2214.
18. Inadera H, Sekiya T, Yoshimura T, Matsushima K.
Molecular analysis of the inhibition of monocyte
chemoattractant protein-1 gene expression by estrogens and xenoestrogens in MCF-7 cells.
Endocrinology. 2000;141:50-59
19. Onai N, Zhang Y, Yoneyama H, Kitamura T,
Ishikawa S, Matsushima K. Impairment of lymphopoiesis and myelopoiesis in mice reconstituted
with bone marrow-hematopoietic progenitor cells
expressing SDF-1-intrakine. Blood. 2000;96:20742080.
20. Sekiya T, Miyamasu M, Imanishi M, Yamada H,
Nakajima T, Yamaguchi M, Fujisawa T, Pawankar R,
Sano Y, Ohta K, Ishii A, Morita Y, Yamamoto K,
Matsushima K, Yoshie O, Hirai K. Inducible expression of a Th2-type CC chemokine thymus- and acti-

vation-regulated chemokine by human bronchial
epithelial cells. J Immunol. 2000;165:2205-2213
21. Shindo T, Kurihara H, Kuno K, Yokoyama H, Wada
T, Kurihara Y, Imai T, Wang Y, Ogata M, Nishimatsu
H, Moriyama N, Oh-hashi Y, Morita H, Ishikawa T,
Nagai R, Yazaki Y, Matsushima K. ADAMTS-1: ametalloproteinase-disintegrin essential for normal
growth, fertility, and organ morphology and function. J Clin Invest. 2000;105:1345-1352.
22. Suzuki T, Hashimoto S, Toyoda N, Nagai S,
Yamazaki N, Dong HY, Sakai J, Yamashita T, Nukiwa
T, Matsushima K. Comprehensive gene expression
profile of LPS-stimulated human monocytes by
SAGE. Blood. 2000;96:2584-2591
23. Tokuda A, Itakura M, Onai N, Kimura H, Kuriyama
T, Matsushima K. Pivotal role of CCR1-positive
leukocytes in bleomycin-induced lung fibrosis inmice. J Immunol. 2000;164:2745-2751
24. Vestergaard C, Bang K, Gesser B, Yoneyama H,
Matsushima K, Larsen CG. A Th2 Chemokine,
TARC, Produced by Keratinocytes May Recruit
CLA+CCR4+ Lymphocytes into Lesional Atopic
Dermatitis Skin. J Invest Dermatol. 2000;115:640646
25. Wada T, Furuichi K, Sakai N, Iwata Y, Yoshimoto K,
Shimizu M, Takeda SI, Takasawa K, Yoshimura M,
Kida H, Kobayashi KI, Mukaida N, Naito T,
Matsushima K, Yokoyama H. Up-regulation of monocyte chemoattractant protein-1 in tubulointerstitiallesions of human diabetic nephropathy. Kidney Int.
2000;58:1492-1499
26. Wada T, Furuichi K, Sakai N, Iwata Y, Yoshimoto K,
Shimizu M, Kobayashi K, Mukaida N, Matsushima K,
Yokoyama H. A new anti-inflammatory compound,
FR167653, ameliorates crescentic glomerulonephritis
in Wistar-Kyoto rats. J Am Soc Nephrol.
2000;11:1534-1541
27. Yamamoto J, Adachi Y, Onoue Y, Adachi YS, Okabe
Y, Itazawa T, Toyoda M, Seki T, Morohashi M,
Matsushima K, Miyawaki T. Differential expressionof the chemokine receptors by the Th1- and Th2type effector populations within circulating CD4+ T
cells. J Leukoc Biol. 2000;68:568-574.
28. Zhang Y, Wang Y, Ogata M, Hashimoto S, Onai N,
Matsushima K. Development of dendritic cells in
vitro from murine fetal liver-derived lineage phenotype-negative c-kit(+) hematopoietic progenitor cells.
Blood. 2000;95:138-146
29. Furuichi K, Wada T, Iwata Y, Sakai N, Yoshimoto K,
Shimizu M, Kobayashi K, Takasawa K, Kida H,
Takeda S, Matsushima K, Yokoyama H. Upregulationof fractalkine in human crescentic glomerulonephritis. Nephron. 2001;87:314-320.
30. Ishikawa S, Sato T, Abe M, Nagai S, Onai N,
Yoneyama H, Zhang Y, Suzuki T, Hashimoto S,
Shirai T, Lipp M, Matsushima K. Aberrant highexpression of B lymphocyte chemokine(BLC/CXCL13) by C11b+CD11c+ dendritic cells inmurine lupus and preferential chemotaxis of B1 cells
towards BLC. J Exp Med. 2001;193:1393-1402.

14131. Itakura M, Tokuda A, Kimura H, Nagai S, Yoneyama
H, Onai N, Ishikawa S, Kuriyama T, Matsushima K.
Blockade of secondary lymphoid tissue chemokine
exacerbates Propionibacterium acnes-induced acutelung inflammation. J Immunol. 2001;166:2071-2079.
32. Kawasaki S, Takizawa H, Yoneyama H, Nakayama T,
Fujisawa R, Izumizaki M, Imai T, Yoshie O, Homma
I, Yamamoto K, Matsushima K. Intervention of thymus and activation-regulated chemokine attenuates
the development of allergic airway inflammation andhyperresponsiveness in mice. J Immunol.
2001;166:2055-2062.
33. Takayama T, Morelli AE, Onai N, Hirao M,
Matsushima K, Tahara H, Thomson AW.
Mammalian and viral IL-10 enhance C-C chemokine
receptor 5 but down-regulate C-C chemokine receptor 7 expression by myeloid dendritic cells: impact
on chemotactic responses and in vivo homing ability.
J Immunol. 2001;166:7136-7143.
34. Wada T, Furuichi K, Sakai N, Hisada Y, Kobayashi K,
Mukaida N, Tomosugi N, Matsushima K, Yokoyama
H. Involvement of p38 mitogen-activated protein
kinase followed by chemokine expression in crescentic glomerulonephritis. Am J Kidney Dis.
2001;38:1169-1177.
35. Yamashita T, Kaneko S, Hashimoto S, Sato T, Nagai
S, Toyoda N, Suzuki T, Kobayashi K, Matsushima K.
Serial analysis of gene expression in chronic hepatitis C and hepatocellular carcinoma. Biochem Biophys
Res Commun. 2001;282:647-654.
36. Yokochi S, Hashimoto H, Ishiwata Y, Shimokawa H,
Haino M, Terashima Y, Matsushima K. An antiinflammatory drug, propagermanium, may target
GPI-anchored proteins associated with an MCP-1
receptor, CCR2. J Interferon Cytokine Res.
2001;21:389-398.
37. Yoneyama H, Matsuno K, Zhang Y, Murai M, Itakura
M, Ishikawa S, Hasegawa G, Naito M, Asakura H,
Matsushima K. Regulation by chemokines of circulating dendritic cell precursors, and the formation of
portal tract-associated lymphoid tissue, in a granulomatous liver disease. J Exp Med. 2001;193:35-49.
38. Higashi N, Fujioka K, Denda-Nagai K, Hashimoto S,
Nagai S, Sato T, Fujita Y, Morikawa A, Tsuiji M,
Miyata-Takeuchi M, Sano Y, Suzuki N, Yamamoto K,
Matsushima K, Irimura T. The macrophage C-type
lectin specific for galactose/N-acetylgalactosamine isan endocytic receptor expressed on monocytederived immature dendritic cells. J Biol Chem.
2002;277:20686-20693.
39. Inadera H, Dong HY, Matsushima K. WISP-2 is asecreted protein and can be a marker of estrogen
exposure in MCF-7 cells. Biochem Biophys Res
Commun. 2002;294:602-608.
40. Ishikawa S, Nagai S, Sato T, Akadegawa K,
Yoneyama H, Zhang YY, Onai N, Matsushima K.
Increased circulating CD11b+CD11c+ dendritic cells(DC) in aged BWF1 mice which can be matured by
TNF-alpha into BLC/CXCL13-producing DC. Eur J
Immunol. 2002;32:1881-1887.

142

41. Kuno K, Baba C, Asaka A, Matsushima C,
Matsushima K, Hosono R. The Caenorhabditis elegans ADAMTS family gene adt-1 is necessary formorphogenesis of the male copulatory organs. J Biol
Chem. 2002;277:12228-12236.
42. Kurachi M, Hashimoto S, Obata A, Nagai S, Nagahata
T, Inadera H, Sone H, Tohyama C, Kaneko S,
Kobayashi K, Matsushima K. Identification of
2,3,7,8-tetrachlorodibenzo-p-dioxin-responsive genes
in mouse liver by serial analysis of gene expression.
Biochem Biophys Res Commun. 2002;292:368-377.
43. Narumi S, Kaburaki T, Yoneyama H, Iwamura H,
Kobayashi Y, Matsushima K. Neutralization of IFNinducible protein 10/CXCL10 exacerbates experimental autoimmune encephalomyelitis. Eur J Immunol.
2002;32:1784-1791.
44. Onai N, Kitabatake M, Zhang YY, Ishikawa H,
Ishikawa S, Matsushima K. Pivotal role of CCL25(TECK)-CCR9 in the formation of gut cryptopatches
and consequent appearance of intestinal intraepithelial T lymphocytes. Int Immunol. 2002;14:687-694.
45. Sekiya T, Yamada H, Yamaguchi M, Yamamoto K,
Ishii A, Yoshie O, Sano Y, Morita A, Matsushima K,
Hirai K. Increased levels of a TH2-type CC
chemokine thymus and activation-regulated
chemokine (TARC) in serum and induced sputum ofasthmatics. Allergy. 2002;57:173-177.
46. Uekusa Y, Yu WG, Mukai T, Gao P, Yamaguchi N,
Murai M, Matsushima K, Obika S, Imanishi T,
Higashibata Y, Nomura S, Kitamura Y, Fujiwara H,
Hamaoka T. A pivotal role for CC chemokine receptor 5 in T-cell migration to tumor sites induced byinterleukin 12 treatment in tumor-bearing mice.
Cancer Res. 2002;62:3751-3758.
47. Ueno T, Hara K, Willis MS, Malin MA, Hopken UE,
Gray DH, Matsushima K, Lipp M, Springer TA, Boyd
RL, Yoshie O, Takahama Y. Role for CCR7 ligands inthe emigration of newly generated T lymphocytes
from the neonatal thymus. Immunity. 2002;16:205218.
48. Wang Y, Zhang Y, Yoneyama H, Onai N, Sato T,
Matsushima K. Identification of CD8alpha(+)
CD11c(-) lineage phenotype-negative cells in the
spleen as committed precursor of CD8alpha(+) dendritic cells. Blood. 2002;100:569-577.
49. Yoneyama H, Narumi S, Zhang Y, Murai M,
Baggiolini M, Lanzavecchia A, Ichida T, Asakura H,
Matsushima K. Pivotal role of dendritic cell-derived
CXCL10 in the retention of T helper cell 1 lymphocytes in secondary lymph nodes. J Exp Med.
2002;195:1257-1266.
50. Yoshie O, Fujisawa R, Nakayama T, Harasawa H,
Tago H, Izawa D, Hieshima K, Tatsumi Y,
Matsushima K, Hasegawa H, Kanamaru A, Kamihira
S, Yamada Y. Frequent expression of CCR4 in adult
T-cell leukemia and human T-cell leukemia virus
type 1-transformed T cells. Blood. 2002;99:15051511.

Department of
Public Health
Outline and Research Objectives
Public health departments in medical schools in Japan were introduced after the WorldWar II, following the model of the U.S. system of medical education. The Department of
Public Health was established in 1948, in the Faculty of Medicine, University of Tokyo. In
1995, the Department became a part of the Division of Social Medicine, Graduate School of
Medicine, as the result of the shift to a graduate school system in University of Tokyo.
The objectives of the Department are both education and research of public health. The
Department trains graduate and undergraduate students through lectures, seminars, fieldpractice, and laboratory work in public health and occupational medicine, for the degrees ofMedical Doctor (MD), Master of Medical Sciences (MSc), and Doctor of Medical Sciences(DMSc). The Department also provides those lectures related to public health and occupational medicine for undergraduate students in the School of Health Sciences and Nursing.
The Department has conducted research on a wide variety of public health issues,
including health policy and economics, occupational medicine, environmental health, community and clinical epidemiology, behavioral medicine, and so on. In addition, the staff
members of the Department have offered public and occupational health services to the central and local governments, industries, and local communities.
Currently, the Department comprises a professor and chair, an associate professor, a lecturer, an associate, a research resident, several part-time lecturers, and several visiting
research fellows, all of who take part in the education and/or the research of the
Department.

Faculties and Students;
Professor and Chair

Yasuki Kobayashi, MD, DMSc
(since April 2001)
Associate Professor Kazuhito Yokoyama, MD, DMSc
Lecturer
Hajime Sato, MD, MPH, DMSc, DPH
Associate .................................1
Research Resident...................1
Postdoctoral Fellow ................1
Graduate Students ................10 (including 3 entrusted
students)
Research Students...................4
Secretary .................................2

Past Research and Major Accomplishments
1) Health policy and economics
We are interested in the topics of health care system and economics in general. We have performed
and published those studies related to supply anddemand sides of health services in Japan; such as supply and distribution of physicians, the separation ofpharmaceutical dispensing and prescribing in medical
practice, cost studies of outpatient and inpatient services, and the efficiency and equity of the Japan’s
health insurance system.
For the issues of physician supply, we examined

and analyzed the impact of physician manpower policy in the 1970s and 1980s in Japan, through both economic and statistical methods, and showed that geographic mal-distribution of physicians did not
improve despite the growing number of physician
during the period. Also, we projected future demandof physicians by specialty in Japan, since this issue
brought a constant debate among the government,
medical schools, medical providers, and patients. Our
projections could facilitate these arguments.
For Japan’s insurance system, we first showed that
it would be cost-beneficial if an electronic claiming
system were introduced to the system. On the other
hand, the financing system for the elderly health careis really a global issue worldwide, and we also havetackled the issue. Since the late 1980s, we haveshown the effectiveness and efficiency of home carefor the elderly in terms of their quality of life. In thelate 1990s, there have been wide and continuous
debates on the matter nationwide, and finally the
new insurance system, namely the Long-term Care
Insurance for the Elderly has been introduced toJapan since the year 2000. Our study results have definitely facilitated these discussions.
We have examined what factors are relevant toearly discharge of elderly patients. We did a follow-up
survey of hospitalized patients with cerebrovascular143

diseases (CVD) in almost all the hospitals in a prefecture. The study showed that the caregiver’s conditions, including economic ones, as well as thepatient’s conditions, were closely related to earlier
home discharge of the CVD patients.
We have carried on several policy studies in thehealth and environmental fields, especially those frominternational comparative perspectives, such as tobacco smoking control, and patient isolation policy for
Hansen’s disease. These studies have been publishedin some international policy journals.
Overall, we have contributed to the evidence based
health policy in Japan, through numerous empiricalstudies using the methods of epidemiology and healtheconomics.

3) Community and clinical epidemiology
We have done several epidemiological studies incommunity, occupational, or clinical settings, such asactive life expectancy for the elderly, quality of life
assessment in RA patients, work-related diseases, andoutcome studies for various kinds of treatments. Mostof these studies have been carried on in collaboration
with local communities, industries, or clinical departments, and published in various journals.

ceuticals has progressed, however, our study has suggested that the separation itself would not necessarily
lead to the cost containment of pharmaceuticals
because physicians tend to prescribe bland-namepharmaceuticals. Therefore, we suggest that such a
separation policy should be combined with a policy
encouraging the use of generics.
In addition, we study the methods and guidelines
by which health insurance claim data will be used forresearch with securing the patients’ privacy. Such a
study would facilitate expanding the volume of andimproving the quality of health services research inJapan, consequently lead to the improvement of quality of health services in Japan.
For the issues of the financing system worldwide,
we have recently began to study a system for providing the effective treatment for people with HIV/AIDS
in developing countries, since the issue is a global andurgent one both in terms of health and economics.
We have conducted a cost study of providing highly
active antiretroviral therapy (HAART) to AIDS
patients in Khon Kaen Province, Thailand. As a result,
we indicate that a substantial increase of resources
would be necessary to provide HAART to all the adult
AIDS patients under the current universal coveragesystem in Khon Kaen province, and we suggest possible solutions for this financial obstacle. This type ofanalysis would be also useful to assess the financial
implications of providing HAART for public healthsystems worldwide.
For the issues of environmental pollution andhealth effects of chemicals, we have recently started
an international collaborative study on pesticide problem in Malaysia. In the study, the health effects ofpesticide in relation to occupational safety behavior
are studied among tobacco-growing farmers in
Kelantan, Malaysia. So far, the following results are
obtained; (i) Organophosphate and dithiocarbamate
pesticides affect peripheral nerve conduction, whereas
pyrethroid affects postural balance system. (ii)
Nonsmoking while spraying, good-sprayer condition,and changing clothes immediately after spraying prevents occurrence of acute symptoms just after pesticide spray in male farmers; in female farmers, wearing
a hat while spraying significantly prevents the symptoms. These results suggest that sub-clinical healtheffects are caused by pesticide use; the effects could
be prevented by safety handling of pesticides so that
health education would be effective.

Current Research

Future Prospects

Most of the above mentioned research topics aretackled at present, with new perspectives andmethodologies. For the insurance system, recently theseparation of prescribing and dispensing of pharma-

Our empirical studies on the health insurance system and health manpower policy in Japan are unique,and our strength. Therefore, we should continue toexpand these kinds of studies and to train profession-2) Occupational medicine and environmental
healthWe have tackled the issues of neurotoxicologic
effects of lead and other heavy metals and solvents,
sarin poisoning, particulate matter monitoring, andhealth effects of pesticides in developing countries aspart of the Alliance for Global Sustainability project.
We have been also interested in the issues of behavioral medicine, such as socioeconomic and psychological aspects of drinking and traffic accidents. For theabove purposes, the Department conducted international collaborative studies in the Faeroes islands ofDenmark (methyl-mercury), Seoul of Korea (lead,chromium and mercury poisonings) and Kota Bahr of
Malaysia (pesticide poisoning). Also, the former chairman of the Department (Araki), as Chairman of the
Scientific Committee on Neurotoxicology andPsychophysiology of the International Commission on
Occupational Health (ICOH), promoted symposia at
the ICOH 2000 Congress in Singapore. The
Department also held the 48th Annual Meeting of theJapanese Society of Occupational Medicine andTraumatology in Tokyo in November 2000.

144

als in such fields, and hope to facilitate and contributeto improving our health care system. Furthermore,international collaborative studies for pubic health arealso a tradition of the Department as well as its
strength in terms of research. We would also like tocontinue such collaborative studies on health of andhealth care for disadvantaged people, such as thepoor, the frail elderly, people with HIV/AIDS, andunderprivileged workers. In addition, since there are
growing needs for epidemiological studies in occupational and clinical settings, we should expand outcome studies and clinical epidemiology, in collaboration with clinical departments. In order to maintainand promote health for all the people in Japan and in
the world, it is essential to promote both theoreticaland empirical studies in public health. The
Department is making every effort for this purpose
through domestic, international and interdisciplinary
collaborative researches.

Research Grants
1. 2001-03 Research Grants from Ministry of Education,
Culture, and Sports. Study on payment system forthe checking functions preventing adverse events ofpharmaceuticals in the separation of prescribing anddispensing. (Kobayashi Y)
2. 2001-03 Health Sciences Research Grants (Researchon Policy Planning and Evaluation) from the Ministryof Health, Labor and Welfare of Japan. Study on use
and security of insurance claim data in Japan’s healthinsurance system. (Kobayashi Y)
3. 2001-04 Grant for Health Cooperation Research from
the Ministry of Health, Labor and Welfare of Japan.
Study on a financing system for providing the effective treatment for people with HIV/AIDS in developing countries. (Kobayashi Y)
4. 2001-03 Research Grants from Ministry of Education,
Culture, and Sports. Study on risk assessment ofsubclinical neuro-behavioral effects of environmental
pollutants. (Yokoyama K)
5. 2001-04 Research Grants from Ministry of Education,
Culture, and Sports. International collaborative study
on health effects of pesticides and their safe use.
(Yokoyama K)

Select Publications
Health policy and economics
1 Kobayashi Y, Yano E. Structure, process, effectiveness and efficiency of the check and review system
in Japan’s health insurance. Health Policy 19: 229244, 1991.
2 Kobayashi Y, Takaki H. Geographic distribution ofphysicians in Japan. Lancet 340: 1391-1393, 1992.

3

Kobayashi Y, Reich MR. Health care financing for theelderly in Japan. Social Science & Medicine 37: 343353, 1993.
4 Kobayashi Y, Chiba Y, Hikita K, Kusumoto K.
Collaboration between Japan and China towards
poliomyelitis eradication [letter]. Lancet 1995; 345:
455.
5 Tamiya, N., Araki, S., Kobayashi, Y., Yamashita, K.,
Murata, K., Yano, E.: Gender difference in the utilization and use’s characteristics of community rehabilitation programs for cerebrovascular disease patientsin Japan. International Journal for Quality in HealthCare 8: 359-366,1996.
6 Ishizaki T, Kobayashi Y, Tamiya N. The role of geriatric intermediate care facilities in long-term care forthe elderly in Japan. Health Policy 43:141-151, 1998.
7 Sato, H.: Policy and politics of smoking control inJapan. Social Science & Medicine 49: 581-600, 1999.
8 Sato, H.: The advocacy coalition framework and thepolicy process analysis: The case of smoking control
in Japan. Policy Studies Journal 27: 28-44, 1999.
9 Sato, H., Araki, S.: Yields and daily consumption ofcigarettes in Japan in 1969-1996. Journal of
Epidemiology 10: 7-15, 2000.
10 Sato, H., Araki, S., Yokoyama, K.: Policy functions ofsmoking control in Japan: Government and social
interests in smoking control in Japan. Environmental
Health and Preventive Medicine 4 : 156-164, 2000.
11 Voorhees, A.S., Araki, S., Sakai, R., Sato, H.: An ex
post cost-benefit analysis of the nitrogen dioxide air
pollution control program in Tokyo. Journal of the
Air & Waste Management Association 50: 391-410,
2000.
12 Sato, H., Araki, S., Yokoyama, K.: Influence ofmonopoly privatization and market liberalization on
smoking prevalence in Japan: Trends of smoking
prevalence in Japan in 1975-1995. Addiction 95:
1089-1098, 2000.
13 Voorhees, A. S., Sakai, R., Araki, S., Sato, H., Otsu,
A.: Cost benefits analysis of nitrogen dioxide control
programmes: A case study of Chiyoda-ku, Tokyo.
Journal of Environmental Planning and Management
44 : 149-165, 2001.
14 Asano, N., Kobayashi, Y., Kano, K.: Issues of interventions aimed at preventing prospective surplus ofphysicians in Japan. Medical Education 35: 488-494,
2001.
15 Tamiya, N., Kobayashi, Y., Murakami, S., Sasaki, J.,
Yoshizawa, K., Otaki, J., Kano, K.: Factors related tohome discharge of cerebrovascular disease patients,
one-year follow-up interview survey of caregivers of
hospitalized patients in 53 acute care hospitals inJapan. Archives of Gerontology and Geriatrics 33:
109-121, 2001.

(2) Occupational medicine and environmental
health16 Hayashi T, Kobayashi Y, Yamaoka K, Yano E. Effectof overtime work on 24-hour ambulatory blood pres-145

sure. Journal of Occupational and Environmental
Medicine 38: 1007-1011, 1996.
17 Tanigawa, T., Araki, S., Nakata, A., Araki, T.,
Sakuraai, S.: Decreases in subpopulations of T lymphocytes and natural killer cells in the blood ofretired chromate workers. International Journal of
Occupational and Environmental Health 2: 222225,1996.
18 Sata, F., Araki, S.,: Adjustment of creatinine-adjusted
value to urine flow rate in lead workers. Archives ofEnvironmental Health 51: 329-333, 1996.
19 Kawakami, N., Araki, S., Kawashima, M., Masumoto,T., Hayashi, T.: Effects of work-related stress reduction on depressive symptoms among Japanese bluecollar workers. Scandinavian Journal of Work,
Environment Health 23: 54-59, 1997.
20 Yokoyama, K., Araki, S., Murata, K., Morita, Y.,
Katsuno, N., Tanigawa, T.,Mori, N., Yokota, J., Ito,
A., Sakata, E.: Subclinical vestibulo-cerebellar, anterior cerebellar lobe and spinocerebellar effects in lead
workers in relation to concurrent and past exposure.
Neurotoxicology 18: 371-380, 1997.
21 Yokoyama, K., Araki, S., Murata, K., Nishikitani, M.,
Nakaaki, K., Yokota, J., Ito, A., Sakata, E.: Postural
sway frequency analysis in workers exposed to nhexane, xylene, and toluene: Assessment of subclinical cerebellar dysfunction. Environmental Research
74: 110-115, 1997.
22 Sata, F., Araki, S., Sakai, T., Nakata, A., Yamashita,K., Morita, Y., Tanigawa, T., Miki, A.: Immunologicaleffects of CaEDTA injection: Observations in twolead workers. American Journal of Industrial
Medicine 32: 674-680, 1997.
23 Grandjean, P., Weihe, P., White, R.F., Debes, F.,
Araki, S., Yokoyama, K., Murata, K., SÉ”rensen, N.,
Dahl, R., JÉ”rgensen, P.J.: Cognitive deficit in 7-yearold children with prenatal exposure to methylmercury. Neurotoxicology and Teratology 19: 417-428,
1997.
24 Murata, K., Araki, S., Yokoyama, K., Okumura, T.,
Ishimatsu, S., Takasu, N., White, R.F.:
Asymptomatic sequelae to acute sarin poisoning in
the central and autonomic nervous system 6 months
after the Tokyo subway attack. Journal of Neurology
244: 601-606, 1997.
25 Yokoyama, K., Araki, S., Murata, K., Nishikitani, M.,
Okumura, T., Ishimatsu, S., Takasu, N.: A preliminary study on delayed vestibulo-cerebellar effects ofTokyo subway sarin poisoning in relation to genderdifference: Frequency analysis of postural sway.
Journal of Occupational and Environmental Medicine
40:17-21, 1998.
26 Yokoyama, K., Araki, S., Murata, K., Nishikitani, M.,
Okumura, T., Ishimatsu, S., Takasu, N., White, R.F.:
Chronic neuro-behavioral effects of Tokyo subway
sarin poisoning in relation to posttraumatic stressdisorder. Archives of Environmental Health 53: 249256, 1998.
27 Yokoyama, K., Araki, S., Aono, H., Murata, K.:
Calcium disodium ethylene-diaminetetraacetate-146

chelated lead as a predictor for subclinical lead neurotoxicity: Follow-up study on gun-metal foundry
workers. International Archives of Occupational andEnvironmental Health 71: 459-464, 1998.
28 Kim, J.M., Araki, S., Kim, D.J., Takasuka, N.,
Babatoriyama, H., Nir,Z., Khachik, F., Shimizu, N.,
Tanaka, Y., Osawa, T., Nishino,H., Tsuda, H.:
Chemopreventive effects of carotenoids and curcumins on mouse colon carcinogenesis after 1,2dimethylhydrazine initiation. Carcinogenesis 19: 8185, 1998.
29 Murata, K., Weihe, P., Renzoni, A., Debes, F.,
Vasconcelos, R,. Zino, F., Araki, S., JÉ”rgensen, P.J.,
White, R. F., Grandjean, P.: Delayed evoked potentials in children exposed to methylmercury fromseafood. Neurotoxicology and Teratology 21: 343348, 1999.
30 Zhang, J., Araki, S., Sato, H., Yokoyama, K.:
Psychobehavioral effects of divorced mother-headed
one-child families on children in China.
Environmental Health and Preventive Medicine 4:
135-139, 1999.
31 Park, S.H., Araki, S., Nakata, A., Kim, Y.H., Park,
J.A., Tanigawa, T., Yokoyama, K., Sato, H.: Effects ofoccupational metallic mercury vapor exposure on
suppressor-inducer (CD4+CD45RA+) T lymphocytes
and CD59+CD16+ natural killer cells. International
Archives of Occupational and Environmental Health73: 537-542, 2000.
32 Kawakami, N., Akaki, K., Shimizu, H., Haratani, T.,
Kobayashi, F., Ishizaki, M., Hayashi, T., Fujita, O.,
Aizawa, Y., Miyazaki, S., Hiro, H., Hashimoto, S.,
Araki, S.: Job strain, social support in the workplace,and hemoglobin A1c in Japanese men. Occupational
and Environmental Medicine 57: 805-809, 2000.
33 Kitamura, F., Araki, S., Suzuki, Y., Yokoyama, K.,
Tanigawa, T., Iwasaki, R. : Assessment of the mutation of the p53 suppressor gene and Ha-and Ki-ras
oncogenes in malignant mesothelioma in relation toasbestos exposure: A study of 12 American patients.
Industrial Health 40: 175-181, 2002.
34 Nordin, R., Araki, S., Sato, H., Yokoyama, K., Muda,
W., Kyi, D. W.: Effects of safety behaviors of pesticide use on occurrence of acute symptoms in maleand female tobacco-growing Malaysian farmers.
Industrial Health 40: 182-190, 2002.
35 Yokoyama, K., Araki, S., Yamashita, K., Murata, K.,
Nomiyana, K., Nomiyama, H., Tao, Y- X., Liu, S-J.,
Sato, H.: Subclinical cerebellar anterior lobe, vestibulo-cerebellar and spinocerebellar afferente effects inyoung female lead workers in China: Computerized
posturography with sway frequency analysis andbrainstem auditory evoked potentials. Industrial
Health 40: 245-253, 2002.

(3) Community and clinical epidemiology
36 Kawakami, N., Iwata, N., Tanigawa, T., Ohga, H.,
Araki, S., Fujikhara, S., Kitamura, T.: Prevalence ofmood and anxiety disorders in working population

in Japan. Journal of Occupational and Environmental
Medicine 38: 899-905, 1996.
37 Yamada, K., Araki, S., Tamura, M., Sakai, I.,
Takahashi, Y., Kashihara, H., Kono, S.: Case-control
study of colorectal carcinoma in situ and cancer in
relation to cigarette smoking and alcohol use inJapan. Cancer Causes and Control 8: 780-785, 1997.
38 Yokoyama, K., Araki, S., Nishikitani, M.: Reasons fordrinking in relation to problem drinking behavior ina sample of Japanese high school students.
International Journal of Behavioral Medicine 6: 135149, 1999.
39 Kawakami, N., Araki, S., Takatsuka, N., Shimizu, H.,
Ishibashi, H.: Overtime, psychosocial working conditions, and occurrence of non-insulin dependent diabetes mellitus in Japanese men. Journal of
Epidemiology and Community Health 53: 359-363,
1999.
40 Yokoyama, K., Araki, S., Matsuoka, K., Mori, N.,
Sato, H., Nakao, M.: Socioeconomic factors affecting
alcohol consumption in Japan. Alcoholism 35: 13-22,
1999.
41 Hotta, Y., Araki, S., Sato, H., Yokoyama, K.: Social
life factors for the mortality from motor vehicle accident. Japanese Journal of Occupational Medicine andTraumatology 48: 483-489, 2000.
42 Ishizaki T, Kobayashi Y, Kai I. Functional transitionsin instrumental activities of daily living among older
Japanese. Journal of Epidemiology 10: 249-254, 2000.
43 Suematsu, Y., Sato, H., Ohtsuka, T., Kotsuka, Y.,
Araki, S., Takamoto, S.: Predictive risk factors ofdelayed extubation in patients undergoing coronary
bypass grafting. Heart and Vessels 15 : 214-220,
2000.
44 Sato, H., Hashimoto, A., Araki, S., Nishibayashi, Y.,
Hoshi, K., Kutsuna, T., Shiino, Y., Ishihara, Y.,
Tsuboi, S., Fujimori, J., Kondo, H., Akizuki, M.,
Moroi, Y., Yoshida, S., Yokoyama, K.: Validity andreliability of a revised Japanese version of the arthritis impact measurement scales version 2 (AIMS2).
Modern Rheumatology 10: 247-255, 2001.
45 Shibata, K., Takamoto, S., Kotsuka, Y., Miyairi, T.,
Morota, T., Ueno, K., Sato, H.: Doppler ultrasonographic identification of the critical segmental arteryfor spinal cord protection. European Journal of
Cardio-Thoracic Surgery 20 : 527-532, 2001.
46 Lee JS, Kawakubo K, Sato H, Kobayashi Y, Haruna Y.
Relationship between total and regional bone mineral density and menopausal state, body compositionand life style factors in overweight Japanese women.
International Journal of Obesity 25: 880-886, 2001.
47 Lee JS, Kawakubo K, Kobayashi Y, Mori K, Kashihara
H, Tamura M. Effects of ten year body weight variability on cardiovascular risk factors in Japanese middle-aged men and women. International Journal of
Obesity 25: 1063-1067, 2001.
48 Ishizaki T, Kai I, Kobayashi Y, Imanaka Y. Functionaltransitions and active life expectancy for older
Japanese living in a community. Archives ofGerontology and Geriatrics 35: 107-120, 2002.

49 Shibata, K., Takamoto, S., Kotsuka, Y., Sato, H.:
Effectiveness of combined blood conservation measures in thoracic aortic operations under deep
hypothermic circulatory arrest. Annals of Thoracic
Surgery 73: 739-743, 2002.
50 Ohyama T, Kobayashi Y, Mori K, Kano K, Sakurai K,
Sato Y. Factors affecting complete resection of gastrictumors by the endoscopic mucosal resection procedure. Journal of Gastroenterology and Hepatology
17:844-848, 2002.

147

Department of
Forensic Medicine
Outline and Research Objectives
Since Professor Kunika Katayama founded our department first in Japan on 1888, we
have performed10258 autopsies by September 30th, 2002. Our precursors have devoted toso many outstanding cases that were paid social attentions. Research objectives are bloodDNA typing, serology, toxicology, forensic pathology, etc. as described below.

Faculties and Students
Professor
Ken-ichi Yoshida, M.D (1999)
Associate Professor Hirotaro Iwase, M.D. (2001)
Associate .................................2
Graduate students...................6
Technical persons ...................4
Secretary .................................2

Past Research and Major Accomplishment
Besides the research on forensic pathology anddetermination of death cause, there have been outstanding accomplishments in basic medical field since
Professor Mita, S., the founder of the Department of
Immunology. Professor Furuhata, T. was outstanding
in the study on the inheritance of ABO blood group.
Professor Ueno, S. discovered complements, while hestudied on medical law. Professor Miki was famousfor blood typing. Professor Ishiyama introduced DNA
typing in the forensic practice, while encouraged
forensic pathology. Professor Takatori, T. was famousin the autopsy and research on the Sarin murdercases, while his research on lipid has been inherited
today.

Current Research
1) Reactive oxygen generation, and Lipid peroxidation and their implication in the pathogenesis:
We identified several lipid species that are generated in the presence of heme-proteins and associatedwith peroxidation. These lipids may be associatedwith the pathogenesis caused by oxidative stresses
that are caused by ischemia-reperfusion (myocardialor brain infarction) or various drugs or toxins(amphetamines, carbon monoxide, agricultural medicines etc.). We undertake to clarify the molecularmechanism of the injury or the cell death due to thesepathogens with reference to lipid peroxidation. Of
these, 4-hydroxynonenal (HNE) is rapidly induced in
the endothelium and sarcolemma of the infarcted

148

myocardium.
Carbon monoxide (CO) is the most common causeof death due to intoxication, and is often supposed toaggravate fatality due to ischemic heart diseases (IHD)or cause delayed neuronal death. CO protects cardiomyogenic cells against ischemia and alleviates reactive oxygen (ROS) generation. By contrast, in the invivo experiments in the rat, CO plus hypoxia induces
necrotic cortical neuronal death, which is associatedwith the enhanced production of lipid peroxide.
Hypothermia reduced the neuronal injury and lipid
peroxide generation, suggesting the implication oflipid peroxidation in the neuronal death induced byCO.
2) The molecular mechanism of the injury and cell
death of myocardium or brain after ischemiareperfusion or toxic substances.
More than two thirds of the causes of unusual
death are ischemic heart or brain diseases. Most ofsuch death occurs within an hour of the onset of thesymptoms. Because it takes more than several hours
from the onset of symptoms for the myocardium toshow distinct histological changes (coagulation necrosis), it is mandatory to establish a diagnostic method
for the early ischemia. To this end, we have to understand the molecular basis of the evolution of theischemic heart or brain disease. We have clarified on
the roles of proteases or intracellular signaling molecules in the pathogenesis of ischemic diseases andtoxic cell injuries. 1) Brief ischemic (e.g. 10min) followed by reperfusion induces proteolysis of cytoskeletal proteins fodrin and ankyrin by Ca2+-dependent
protease calpain, which causes contractile dysfunctionin the isolated heart. This is the earliest irreversible
change so far documented. 2) Protein Kinase C (PKC)
isoforms translocate to membrane or nucleus during
ischemia. Nitric oxide (NO) mediates the translocationof PKC-α, δ, and ε, which protects myocardium
against reperfusion injury. 3) Ischemia-reperfusioncauses sequential activation of PI3 kinase-PKC-ζ-MEKMAP kinase- c-fos , thereby preventing apoptotic
death. This study disclosed that MAP kinase translo-

cates to the nucleus independent of phosphorylation,
but is activated by phosphorylation through MEK activation during postischemic reperfusion in the nucleus,in contrast with the paradigm. 4) Hypoxia under normoglycemia causes autophagic cell death through acidosis through PI3 kinase-mediated activation of glycolysis, explaining the myocardial death under pulmonary hypoventilation or hypoxic hypoxia. 5)
Angina attacks often attenuate the injury due to subsequent myocardial infarction (MI). This phenomenon, ischemic preconditioning (IP), is reproduced in
rat coronary occlusion model of acute MI. IP induces
endothelial NO synthase (eNOS) and collateral vasodilation in the early MI, whereas IP induces angiogenesis in late MI through PKC- ε activation and VascularEndothelium Growth Factor (VEGF) up-regulation.
The both salvage myocardium in MI. 6) Connexin(Cx)-43 undergoes proteolysis during early MI
through lysosomal and proteasomal proteases independently of calcineurin-mediated proteolysis. IP
attenuates the Cx43 proteolysis in early MI. Thesefinding explain the arrhythmias in early MI and its
prevention by IP. 7) Emotional stress evoked byimmobilization of rat causes activation of MAP
kinase-c-fos pathway. 8) Methamphetamine (MAH)
induces hyperthermia through symphathetico-adrenal
activation and enhanced skeletal muscle metabolism,
which is blocked by a sarcoplasmic reticulum Ca2+release blocker dantrolene. Repeated intermittent
administration of MAH causes enhanced cardiovascular, thermal, and behavioral responses to MAH or
emotional stress. We are willing to apply the findingsof the experimental studies to exploit a new method
for the diagnosis of ischemic or other disease states.
3) The study on the legal aspects of forensic medicine.
Forensic practices are associated with various legal
or social problems that ought to be solved, though
there have been research on such issues. On the other
hand, those who provide expert opinion in the civil orlegal litigation were far less than those required.
There is dissociation in the thought-ways between
medical doctors and lawyers, which can seriously violate the human right of persons concerned as well as
causes the shortage of the expert witnesses. The
courts have made effort to increase the numbers ofexpert witnesses for the litigation, though no adequate. On the other hand, the expression or the wayof the expert witness may affect the judgment in thelitigation or prosecution, as suggested by the questionnaire study.
The "reportable unusual deaths" in the guideline
proposed by Japanese Society of Legal Medicine
includes the unexpected death during or shortly after
any medical practice. According to the Doctorﾕs Act

in Japan, doctors must report unusual death cases tothe police. The police (prosecutors) demand autopsy
under the guidance of superintendents. In England,coroners request autopsy after inquiry with the police,
relatives and forensic pathologists. Medical accidents
are largely autopsied in England whereas, in Japan,
medical accidents are rarely reported nor. In Tokyoand few other districts, medical examiners decide
autopsy of 1/4~1/3 of unusual deaths, but autopsies inother districts are mostly conducted by police judgment in search for a possible crime. It should be
reminded that death cause tells criminality and the
responsibility only if the cause is diagnosed exactly by
autopsy performed under proper judgment and that
autopsy is often effective to clarify the cause of unexpected deaths. Recently in Japan, unexpected deaths
in hospitals have come to be increasingly suspected as
caused by malpractice. For lack of a proper system toanswer for complaints from patientsﾕ side, the relatives sometimes go to the police. Though most cases
are asked only for civil liability, the police investigate
the hospital in question and this makes doctors hesitate to report. Prosecutors judge criminality on autopsy operatorﾕs expert opinion, whereas they insist that
autopsy results should not be disclosed to protect privacy of investigation. Thus, an insufficient information on patientﾕs side explains to a great extent why
nearly 700 relatives sue doctors every year in Japan.
In 2001, Japan Surgical Society announced to the public and Society of Legal Medicine that patientsﾕ death
during or shortly after operations should not be
accounted as an unusual death reportable to thepolice, but to the independent authority, if required.
Though many cases are unavoidable and not problematic, doctors sometimes neither inform the patientﾕs
side enough to get satisfactory consent for the practice nor explain the necessity of autopsy to clarify
death cause. As accepted in U.K., there should only bereasonable cause to suspect for "unusual death".
Accordingly, although deaths in medical accident may
be sometimes inevitable or unrelated to the malpractice, the doctorﾕs explanation as such for an unexpected death can be hardly accepted by patientﾕs side.
Therefore, independent investigation and autopsy are
required in medical accidents in order to determineand explain the cause of death to patientﾕs side andto disclose and improve the medical practice by doctors. We are willing to devote to set up valid system
for the death investigation in the medical accident
cases.
4) Forensic pathology.
Because each case is unique and diversity is a
important determinant of the death, we try to perform microscopic analyses in as many cases as possible. We undertake to take the advantage of the above-149

mentioned studies in the immunohistochemical
method for the diagnosis of our autopsy cases.

Future Prospective
The correct and fair determination of deathcause in the medico-legal autopsy cases has been themost important mission of us. We must accept very
difficult and socially notable victims and requirementsof expert opinion. We must play this important role
carefully and impatiently, and train young forensic
pathologists.
There has been not so much evolution in theforensic practice except for the DNA identification
technology. This may be the cause of the low overall
activity in the forensic medicine. To open the window
to the future of forensic medicine, we must do every
effort to find new directions of forensic medicine andlet them known to the public as well as medical field.
Actually, there are many medico-legal issues in theclinical medicine, which have manifested as the growing criticisms from the society on the medical malpractice, inadequate risk management of the medical
accidents, etc. In Japan, the death investigation system for the medical accidents and the unexpected
death in hospitals is quite immature, so that it is very
important and urgent to set up an independent organization for the death investigation for such cases. As
forensic pathologists, we have autopsied such cases
and recognize the problems to be addressed. We aregoing to take part in such activity in cooperation withclinicians and civil servants to find and build up thegood system. Additionally, the education on theseissues to the clinicians, citizens, as well as medical
and law students are very important.
The legal systems for death investigation of"unusual death", prosecution and litigation haveinherent drawbacks, which require improvement. We
have taken effort to inform on the problems to persons concerned, but it is far from what is ought to be.
The 15~20% of total deaths are "unusual deaths",
which requires death investigation. The misdiagnosesof the "unusual deaths" by clinicians, and police surgeons, are police officers have been manifested due tothe inherent inadequacy of the system and poor education of those who investigate. Because there aresome typical patterns of misdiagnoses and poor management of those cases, we must educate on theseissues to students, medical service personnel, and thesociety.

Research Grant
Grant-in-aid from Monbusho (B) for Yoshida et al.
1) A survey for the myocardial proteins and genes
induced by ischemia or psychological stress and its

150

application to the forensic practices (1998~1999)
10,200,000 yen
2) Identification and path-physiological study of novel
peroxidized fatty acids formed in the tissue after
ischemia or various intoxication and the exploitationof the new diagnostic method (2000~2001)
14,800,000 yen
3) Research on the mechanism of cellular injury due tonitric oxide or carbon monoxide in tissues undergoing ischemia or shock (2002) 14,000,000 yen

Grant-in-aid from Monbusho (C) for Uemura,
Yoshida et al.
4) Research on the intracellular stress response in theamphetamine-induced neuro-degeneration (1999~
2000) 2,500,000 yen
5) Research on the molecular mechanism of promotion
and protection of cell death by carbon monoxide(2002) 2,700,000 yen

Select Publications
1. Yoshida K, Inui M, Harada K, Saido TC, Sorimachi Y,
Ishihara T, Kawashima S, Sobue K. Reperfusion of
rat heart after brief ischemia induces proteolysis ofcalspectin(nonerythroid spectrin or fodrin) by calpain. Circ Res. 77, 603-10, 1995.
2. Sorimachi Y, Harada K, Yoshida K. Involvement of
calpain in postmortem proteolysis in the rat brain.
Forensic Sci Int. 15, 81, 165-74.
3. Yoshida K, Hirata T, Akita Y, Mizukami Y,
Yamaguchi K, Sorimachi Y, Ishihara T, Kawashiama
S. Translocation of protein kinase C-alpha, delta andepsilon isoforms in ischemic rat heart. Biochim
Biophys Acta. 1996 Oct 7;1317(1):36-44.
4. Sorimachi Y, Harada K, Saido TC, Ono T, Kawashima
S, Yoshida K. Downregulation of calpastatin in rat
heart after brief ischemia and reperfusion. J Biochem(Tokyo). 1997 Oct;122(4):743-8.
5. Kitakaze M, Funaya H, Minamino T, Node K, Sato
H, Ueda Y, Okuyama Y, Kuzuya T, Hori M, Yoshida
K. Role of protein kinase C-alpha in activation ofecto-5 -nucleotidase in the preconditioned canine
myocardium. Biochem Biophys Res Commun 1997
Oct 9;239(1):171-5
6. Fukuda S, Harada K, Kunimatsu M, Sakabe T,
Yoshida K. Postischemic reperfusion induces alphafodrin proteolysis by m-calpain in the synaptosome
and nucleus in rat brain. J Neurochem. 1998
Jun;70(6):2526-32.
7. Yoshida K, Harada K. Proteolysis of erythrocyte-type
and brain-type ankyrins in rat heart after postischemic reperfusion. J Biochem (Tokyo). 1997
Aug;122(2):279-85.
8. Harada K, Fukuda S, Kunimoto M, Yoshida K.
Distribution of ankyrin isoforms and their proteolysis after ischemia and reperfusion in rat brain. J
Neurochem. 1997 Jul;69(1):371-6.
9. Harada K, Sorimachi Y, Yoshida K. Proteolysis ofankyrin and Na+/K+-ATPase in postmortem rat brain:

is calpain involved? Forensic Sci Int. 1997 Apr
18;86(1-2):77-85.
10. Mizukami Y, Yoshioka K, Morimoto S, Yoshida K. A
novel mechanism of JNK1 activation. Nuclear
translocation and activation of JNK1 during ischemia
and reperfusion. J Biol Chem. 1997 Jun
27;272(26):16657-62.
11. Mizukami Y, Yoshida K. Mitogen-activated protein
kinase translocates to the nucleus during ischaemia
and is activated during reperfusion. Biochem J. 1997
May 1;323 ( Pt 3):785-90.
12. Mizukami Y, Hirata T, Yoshida K. Nuclear translocation of PKC zeta during ischemia and its inhibition
by wortmannin, an inhibitor of phosphatidylinositol
3-kinase. FEBS Lett. 1997 Jan 20;401(2-3):247-51.
13. Kawamura S, Yoshida K, Miura T, Mizukami Y,
Matsuzaki M. Ischemic preconditioning translocates
PKC-delta and -epsilon, which mediate functional
protection in isolated rat heart. Am J Physiol. 1998
275(6 Pt 2):H2266-71.
14. Kawata Y, Mizukami Y, Fujii Z, Sakumura T, Yoshida
K, Matsuzaki M. Applied pressure enhances cell proliferation through mitogen-activated protein kinase
activation in mesangial cells. J Biol Chem. 1998 Jul
3;273(27):16905-12.
15. Makisumi T, Yoshida K, Watanabe T, Tan N,
Murakami N, and Morimoto A. Sympatho-adrenal
involvement in methamphetamine-induced hyperthermia through skeletal muscle hypermetabolism.
Eur J Pharmacol 363, 107-112, 1998.
16. Shama KMA, Suzuki A, Harada K, Fujitani N,
Kimura H, Ohno S, Yoshida K. Transient up-regulation of myotonic dystrophy protein kinase-binding
protein, MKBP, and HSP27 in the neonatal myocardium.Cell Struct Funct 1999; 24; 1-4.
17. Yoshida K, Mizukami Y, Kitakaze M. Nitric oxide
mediates protein kinase C isoform translocation in
rat heart during postischemic reperfusion. Biochim
Biophys Acta 1999; 1453; 230-8.
18. Harada K, Ichiyama T, Ikeda H, Yoshida K. A fatalcase of oral ingestion of benzine. Am J Forensic
Med Pathol 1999; 20; 84-9.
19. Harada K, Maekawa T, Shama KMA, Yamashima T,
Yoshida K. Translocation and down-regulation ofprotein kinase C-α,-β, and -γisoforms during
ischemia-reperfusion in rat brain. J Neurochem 1999;
72; 2556-64.
20. Yoshida K, Aki T, Harada K, Shama KMA, Kamoda
Y, Suzuki A, Ohno S. Translocation of HSP27 and
MKBP in ischemic heart. Cell Struct Funct 1999; 24;
181-5.
21. Harada K, Asa K, Imachi T, Yamaguchi Y, Yoshida K.
Centipede inflicted postmortem injury. J Forensic Sci
1999; 44; 849-50.
22. Ueyama T, Yoshida K, Senba E. Emotional stressinduces immediate-early gene expression in rat
heart via activation of alpha- and beta-adrenoceptors.
Am J Physiol 1999; 277; H1553-61.
23. Sakurada K, Iwase H, Takatori T, Nagao M,
Nakajima M, Niijima H, Matsuda Y, Kobayashi M.

Identification of cis-9,10-methylenehexadecanoic
acid in submitochondrial particles of bovine heart.
Biochim Biophys Acta 1999; 1437; 214-22.
24. Niijima H, Nagao M, Nakajima M, Takatori T,
Matsuda Y, Iwase H, Kobayashi, M. Sarin-like,soman-like organophosphorous agents activate PLC
gamma in rat brains. Toxicol Appl Pharmacol 1999;
156; 64-9.
25. Kobayashi M, Takatori T, Nakajima M, Saka K,
Iwase H, Nagao M, Niijima H, Matsuda Y. Does thesequence of onset of rigor mortis depend on the proportion of muscle fiber types and on intra-muscularglycogen content? Int J Legal Med 1999; 112; 167-71
26. Kobayashi M, Sakurada K, Nakajima M, Iwase H,
Hatanaka K, Matsuda Y, Ikegaya H, Takatori T. A
keyhole lesion gunshot wound in an adipocere
case. Legal Med 1999; 1; 170-3.
27. Ikegaya H, Iwase H, Hatanaka K, Sakurada K,
Matsuda Y, Kobayashi M, Takatori, T: Postmortem
changes in cytochrome c oxidase activity in variousorgans of the rat and in human heart. Forensic Sci
Int 108, 181-186, 2000
28. Mizukami, Y, Kobayashi, S, Uberall, F, Hellbert, K,
Kobayashi, N and Yoshida, K.: Nuclear mitogen-activated protein kinase activation by protein kinase C
zeta during reoxygenation after ischemic hypoxia. J
Biol Chem, 275, 19921-19927, 2000.
29. Ueyama, T, Yoshida, K and Senba, E.: Stress-induced
elevation of the ST segment in the rat electrocardiogram is normalized by an adrenoceptor blocker.
Clinical Exp Pharmacol Physiol, 27, 384-386, 2000.
30. Iwase, H., Sakurada, K., Hatanaka, K., Kobayashi, M.
and Takatori, T.: Effect of cytochrome c on thelinoleic acid-degrading activity of porcine leukocyte
12-lipoxygenase. Free Radical Biol Med, 28, 912-919,
2000.
31. Nakajima, M., Nagao, M., Iwasa, M., Monma-Ohtaki,J., Maeno, Y., Koyama, H., Seko-Nakamura, Y., Isobe,
I. and Takatori, T.: Related Articles：Purificationand characterization of diquat (1, 1 -ethylene-2,
2 -dipyridylium)- metabolizing enzyme fromparaquat-resistant rat liver cytosol. Toxicology, 154,
55-66, 2000.
32. Shiraishi, K, Naito, K. and Yoshida, K. Inhibition of
calpain but not caspase protects the testis against
injury after experimental testicular torsion of rat.
Biol. Reproduct, 63, 1538-1548, 2000.
33. Tsuji T, Ohga Y, Yoshikawa Y, Sakata S, Abe T,
Tabayashi N, Kobayashi S, Kohzuki H, Yoshida KI,
Suga H, Kitamura S, Taniguchi S, Takaki M. Rat cardiac contractile dysfunction induced by Ca2+ overload: possible link to the proteolysis of alphafodrin.Am J Physiol Heart Circ Physiol 2001
281(3):H1286-9
34. Uemura K, Harada K, Sadamitsu D, Tsuruta R,
Takahashi M, Aki T, Yasuhara M, Maekawa T,
Yoshida K. Apoptotic and necrotic brain lesions in a
fatal case of carbon monoxide poisoning. Forensic
Sci Int 2001; 116, 213-9.

15135. Uemura K, Yoshida K. Seat belt induced transectionof trachea in a child held on laps of adult upon car
diving. J Forensic Sci 2001; 46, 714-6.
36. Sanada S, Kitakaze M, Papst PJ, Hatanaka K,
Asanuma H, Aki T, Shinozaki Y, Ogita H, Node K,
Takashima S, Asakura M, Yamada J, Fukushima T,
Ogai A, Kuzuya T, Mori H, Terada N, Yoshida K,
Hori M. Role of phasic dynamism of p38 mitogenactivated protein kinase activation in ischemic preconditioning of the canine heart. Circ Res. 2001 Feb
2;88(2):175-80.
37. Kawata H, Yoshida K, Kawamoto A, Kuroiwa H,
Takase E, Sasaki Y, Hatanaka K, Kobayashi M,
Ueyama T, Hashimoto T, Dohi K. Ischemic preconditioning upregulates vascular endothelial growth factor mRNA expression and neovascularization via
nuclear translocation of protein kinase C e in the rat
ischemic myocardium. Circ Res 2001; 88: 696-704.
38. Hatanaka K, Ikegaya H, Takase I, Kobayashi M,
Iwase H, Yoshida K. Immobilization stress-induced
thymocyte apoptosis in rats. Life Sci 2001; 69: 15565.
39. Ikegaya H, Iwase H, Hatanaka K, Sakurada K,
Yoshida K, Takatori T. Diagnosis of cyanide intoxication by measurement of cytochrome c oxidase activity. Toxicol Lett. 2001; 119: 117-23.
40. Yamada Y, Kobayashi M, Takatori T, Nagao M, Ohira
H, Yamamoto I, Ohtani S. Possibility of paternity
testing using RFLP analysis of a very small amountof material. J Forensic Odontostomatol 2001; 19: 1-4.
41. Yamada Y, Takatori T, Nagao M, Iwase H, Kuroda N,
Yanagida J, Shinozuka T. Expression of paraoxonase
isoform did not confer protection from acute sarin
poisoning in the Tokyo subway terrorist attack. Int J
Legal Med. 2001; 115: 82-4.
42. Aki T, Mizukami Y, Oka Y,Yamaguchi Y, Uemura K,
Fujimiya T, Yoshida K. Phosphoinositide 3-kinase
accelerates necrotic cell death during hypoxia.
Biochem J 2001; 358, 481-7.
43. Kudo R, Adachi J, Uemura K, Maekawa T, Ueno Y,
Yoshida K. Lipid peroxidation in the rat brain after
CO inhalation is temperature dependent. Free
Radical Biol Med 2001; 31, 1417-23.
44. Iwase H, Kobayashi M, Nakajima M, Takatori T. The
ratio of insulin to C-peptide can be used to make aforensic diagnosis of exogenous insulin overdosage.
Forensic Sci Int 2001; 115: 123-7.
45. Iwase H, Kobayashi M, Kurata A, Inoue S. Clinicallyunidentified dissection of vertebral artery as a causeof cerebellar infarction. Stroke 2001; 32: 1422-4.
46. wase H, Sakurada K, Ikegaya H, Hatanaka K,
Takeichi H, Kobayashi M, Yoshida K. Thiol-oxidizing
agent diamide and acidic pH enhance lipid peroxidation of rat heart mitochondria and cardiolipincytochrome c complex. IUBMB Life 2001; 51: 39-43.
47. Kobayashi M, Ikegaya H, Takase I, Hatanaka K,
Sakurada K, Iwase H. Development of rigor mortis is
not affected by muscle volume. Forensic Sci Int 2001;
117: 213-9.

152

48. Shiraishi K, Naito K, Yoshida K. Vasectomy impairs
spermatogenesis through germ cell apoptosis mediated by p53-Bax pathway in the rat. J Urology 2001;
166: 1565-1568.
49. Shiraishi K, Yoshida K, Fujimiya T, Naito K.
Activation of mitogen activated protein kinases andapoptosis of germ cells after vasectomy in the rat. J
Urology 2002; 168: 1273-1278.
50. Yoshida K, Kuroki H, Takeichi H, Kawai K. Deathduring surgery in Japan. The Lancet 2002; 360: 805.

Department of Medical
Informatics and Economics
Outline and Research Objectives
In 1983, the Department of Hospital Information Management(Hospital Computer
Center) was established as one of central facilities in the University of Tokyo Hospital. The
major roles have been designing, implementing, and maintaining the hospital information
management system based on up-to-date information technology and improving functionsof clinical information management required in the university hospital. The University
Medical Information Network Center(UMIN Center) was opened in the department as acooperate organization for national medical schools in Japan. The purposes of the UMIN
Center are to provide up-to-date communication environment to healthcare professionals
via the Internet. In 1997, according to the reconstruction of Graduate School, theDepartment of Medical Informatics and Economics was established in the Division of Social
Medicine, Graduate School of Medicine, and the functions of research and education andthe staffs were formally moved from the hospital to the department in the Graduate Schoolof Medicine. However, all the administrative works for providing various services of hospital information management have been conducted by the same staffs as before.
From the responsibilities of the hospital described as above, most of the researches havebeen based on practical development, integration, and implementation of hospital information systems using an innovative information technology. The scope covers standardizationof medical information exchange, evaluation of new technologies for distributed hospital
information systems, development of electronic medical record systems(EMRS) and medical
decision support systems(MDSS), application of the Internet technology in healthcare fields,
and method of knowledge discovery from clinical database. The goal is to contribute toimprovement of quality of care using information technology

Faculties and Students
Professor and Chair Kazuhiko Ohe, M.D., Ph.D. (1997-)
Associate Professors Takahiro Kiuchi, M.D., Ph.D. (forUMIN Center)
Graduate student ....................9
Research student.....................2

Past Research and Major Accomplishments
1) Development and evaluation of a hospital
information system using open standard and HL7(1993-Current)
Although the Health Level Seven(HL7) Version 2.1
was the only proposal for connecting distributed
information systems in a hospital around 1993, it was
no so-called “standard” protocol. Because it was essential for developers of develop large-scale hospital
information systems(HISs) to introduce a standard
communication protocol that could be generally usedin different departmental computer systems and theirterminals, an experience and evaluation of real implementation of HL7 in a large-scale HIS were needed in
the field of medical informatics in the early ‘90s. Ohe

designed a new architecture of client-server-type HIS
using HL7-based messaging protocol between client
terminals and the server over TCP/IP network, which
had never been used in Japan. The innovative HIS
was successfully implemented based on this newarchitecture in the University of Tokyo Hospital in
1995 and the good performance was proved in this
implementation[7]. This first experience of implementing the standard communication protocol or HL7
contributed to move most HISs in Japan from thevender proprietary systems into multi-vender distributed systems using open standard. Subsequent activities related to the open standard brought about foundation of the Association of HL7 Japan(Ohe is thevice-chair of the technical committee), foundation ofJapan Health Information and Communication
Standards Board (HELICS Board; Ohe is the Chair of
the Board),
2) Modeling a semantic structure of clinical information using object-oriented approach and development of HIS using OODB and CORBA(1996Current)
Analyzing and Modeling a semantic structure ofclinical information in terms of information science153are essential tasks to develop both multi-purpose
EMR systems and future intelligent decision support
systems. Object-oriented analysis(OOA) and modeling(OOM) were proposed in the ‘90s as powerful
methods to model a semantic relationship among
information objects in a target domain and they were
gradually applied in various fields as well as medical
information. We recognized the importance of the
OOA/OOM at the early stage, and focused on applying the approach to developing a future intelligent
clinical information systems. At first we analyzed andmade a model of data of ECG, and implemented aretrievalsystemusing
Object-Oriented
Database(OODB)[5,6,10]. The research proved the feasibility and realization of OOA/OOM and OODB system to develop multi-media clinical information systems.
Based on this result, we proposed a new architecture for managing distributed medical information
using OODB and CORBA(Common Objects Request
Broker Architecture). This approach aimed to shareinter-hospital distributed clinical information forimprovement of quality of care and quality of multiinstitutional large-scale clinical researches. In thisresearch, we have been developing CORBA-based
EMR systems in the University of Tokyo
Hospitals[12,18,21].
3) Standardization of medical informationexchange and terminology (1997-Current)
Ohe and the collaborative researchers in otherinstitutes have been concurrently developing a new
standard protocol for exchanging medical informationelectronically among hospitals in collaboration with
the Association of HL7-USA /Japan and related working group of ISO/TC215, in which Ohe is the delegateof Japan of WG3; concept representation). We proposed a protocol named “MML(Medical Markup
Language)” based on XML(eXtensible Markup
Language) in the early stage of the work[8], and later
we proposed a new protocol named “MERIT-9” basedon integration of XML, HL7, DICOM, and other defacto standard through our experiences of implementing the previous protocol[14,16]. This new protocol isin the stage of evaluation in several hospitals in Japan.
In the domain of medical terminology, Japan had
double standards of Japanese clinical disease terminology and all of hospitals in Japan have not decided touse which of them for a long time. Ohe and Kaihara,
who was a former professor of this department, proposed a method of integrating these two Japanesestandard into one unified standard terminology thatis available for general-purpose clinical information
systems and EDI(Electronic Data Interchange) systemof health insurance claim reimbursement. Each termin the unified disease terminology is linked to an ICD-

154

10 code and semantic relations among the terms.
Further a new standard methods to access to a database of the terminology was developed and published
as a open software in 2001 by Ohe and the colleagues.
Most large-scale university hospitals has been moving
to introduce the standard disease terminology since
2002 and Japan Ministry of Health, Labor, andWelfare announced that all of the hospitals must use
this terminology including the codes in EDI.
4) Application of the Internet technology to medical field by the UMIN
Application of the Internet technology to medical
field by the UMIN has been the major topic of theresearches[13,15,17,]. Associate Professor Kiuchi isthe director of the UMIN Center and he has beendeveloping and managing the nation-wide medical
information network sponsored by the Ministry of
Education, Culture, Science, Sports and Technology(MEXT). Using this infrastructure, Kiuchi and his colleagues proposed a standard method of on-line
abstract and paper entry and developed a system forJapanese academic medical societies. Hundreds of thesocieties have been adopting this system and as aresult this methods and the system has become thede-facto standard in Japan. Kiuchi and his colleagues
also developed Internet-based data collection system
for nation-wide clinical and epidemiological research.
The system have been enhancing the scale and theefficiency of such research[22].

Current Research
Most of the researches described above is continuing and currently expanded toward the next stage.
1) Enhancing Standardization of electronic representation of medical information:
Revising and evaluating the “MERIT-9” standard
protocol in harmonization with HL7 ver.3 that is proposed to cover comprehensive clinical information
systems.
2) Implementing large-scale EMR system integrated with order entry system using distributed
object technology:
Based on the evaluation study of CORBA-based
HIS, we are preparing and developing large-scale EMR
system that will be operated from Apr. 2003 in the
University of Tokyo Hospital.
3) Trial of knowledge discovery from large-scale
medical database created by HIS for futureIntelligent decision support system:
We are investigating a method of data mining andknowledge discovery from large-scale medical database[23].

Future Prospects
We will shift to develop an intelligent decision
support system that uses both large-scale clinical database and real-time knowledge discovering process,and for the purpose a new method of data mining
including both natural language processing and temporal data processing should be created. Further, wewant to focus on a method of creating an practical
ontology that is specific to healthcare domain. In
other words, integration of management of clinicaldata and handling of electronic medical knowledge isthe next objectives for our goals.

Research Grants
five recent grants selected:
1) 2000F.Y.-2001F.Y. Health and Labor Sciences
Research Grants of MHLW, Research on HealthTechnology Assessment, Research on developmentof formal electronic representation of logics of Japan
medical fee payment system and a standard softwareof source-code generator for calculating medical fee.
Head Researcher.
2) 2002F.Y Health and Labor Sciences Research Grants of
MHLW, Research on Policy Planning and Evaluation,
Investigation of DRG for inpatients with acute diseases. Co-researcher.
3) 2001F.Y.-2002F.Y. Grants-in-Aid for Scientific
Research of MEXT, Scientific Research-C,
Development of a method to discover similar cases
from clinical database using semantic relationshipamong medical terms, Head researcher.
4) 2001F.Y-2002F.Y. Health and Labor Sciences Research
Grants of MHLW, Research on Policy Planning and
Evaluation, A study on roles of Health Insurance
Agency, Head researcher.
5) 1999F.Y.-2001F.Y. Health and Labor Sciences
Research Grants of MHLW, Research on HealthTechnology Assessment, A study on modeling
healthcare information domain. Head Researcher.

Select Publications
1) Ohe,K., Kaihara,S.: An Object -Oriented Model ofPhysicians’ Strategy at First Encounters - An
Approach to sharing Distributed Knowledge-bases.
K.C.Lun et al.(eds), Elsevier Science Publishers,
B.V.(North-Holland), MEDINFO92: 434-439, 1992.
2) Kaihara S., Ohe K., Sakurai T. and Kiuchi T.: HIS andResearch. Hospital Information Systems: ScopeDesign-Architecture, A.R. Bakker et al. (eds), Elsevier
Science Publishers B.V.(North-Holland), 97-101,
1992.
3) T. Koyama, K. Ohe: Building a Common Knowledge
Base for Internal Medicine. Proceedings ofKorea/Japan Joint Conference on Expert Systems:
876-888,Seoul (Korea), 1993.
4) K.Ohe, S.Kaihara, K.B.Ishikawa, T.Hishiki, T.Nagase,
T.Sakurai: Hospital Information system and theInternet. K.Chon (ed), Proceedings of theInternational Networking Conference INET’95, 10331036, Internet Society, 1995.
5) C.Wang, K.Ohe, T.Sakurai, T.Nagase, S.Kaihara: An
ECG Storage and Retrieval System Embedded in
Client Server HIS Utilizing Object-Oriented DB.
Journal of Medical Systems, 20(1), 35-43, 1996.
6) C.Wang, K.Ohe, T.Sakurai, T.Nagase, S.Kaihara:
Object-oriented Analysis and Design of an ECG
Storage and Retrieval System Integrated with an HIS.
Methods of Information in Medicine, 35(1), 35-40,
1996.
7) K.Ohe, S.Kaihara: Implementation of HL7 to ClientServer Hospital Information System(HIS) in the
University of Tokyo Hospital. Journal of Medical
Systems, 197-205, Vol.20, No.4, 1996.
8) Yoshihara H., Ohe K., Ohashi K., Yamamoto R.,
Yamazaki S., Hirose Y., et.al.: Standardization of
Exchange Procedures of Clinical Information, and an
Experiment of Clinical Data Exchange Using Medical
Markup Language (MML). Journal of Japan Medical
Informatics, 17(3)Suppl., 203-207,1997.
9) Hishiki T.,Ohe K., Kaihara S.: Extraction of Clinical
Information from Narrative Medical Records Using
Natural Language Processing Journal of Japan
Medical Informatics, 17(3)Suppl.,1997.
10) Wang C, Ohe K, Kaihara S: Dynamic link between
ECG and clinical data by a CORBA-based query
engine and temporal mapping. Proceedings of AMIA
Annual Fall Symposium,27-31,1997.
11) K.Miyo, K.Ohe: SGML-based Construction andAutomatic Organization of Comprehensive Medical
textbook on the Internet. Proceedings of MEDINFO98, B.Cesnik etal.(Eds), Amsterdam:IOS press,145149,1998.
12) K.Ohe: A Hospital Information System based on
Common Object Request Broker Architecture(CORBA) for Exchanging Distributed Medical Objects- an approach to future environment of sharing
healthcare information. Proceedings of MEDINFO98,
B.Cesnik etal.(Eds), Amsterdam:IOS press,962964,1998.
13) T. Kiuchi, T. Sakurai, K. Ohe, Y. Ohashi, S. Kaihara:
University Medical Information Network - Past,
Present, and Future. Proceedings of MEDINFO98,
B.Cesnik etal.(Eds), Amsterdam:IOS press,420424,1998.
14) M. Kimura, K.Ohe, H. Yoshihara, Y. Ando, F.
Kawamata, T. HIshiki, et.al.: Patient Information
Exchange Guideline MERIT-9 using Medical Markup
Language MML. Proceedings of MEDINFO98,
B.Cesnik etal.(Eds), Amsterdam:IOS press,433437,1998.
15) H． Yamakami， T.Kiuchi, T.Nagase， K.Ohe,
S.Kaihara, T.Sakurai: Development and Trial operation of a World Wide Web-based data entry system

155

for the collection of statistical data on the management of the national university hospitals in Japan.
Medical Informatics,23(1),19-29,1998.
16) M. Kimura, K. Ohe, H. Yoshihara, Y. Ando, F.
Kawamata, T. Hishiki, et.al.: MERIT-9; a patient
information exchange guideline using MML, HL7,
and DICOM. International Journal of Medical
Informatics, 59-68,51(1),1998.
17) T.Kiuchi, K.Ohe, S.Kaihara: Using WWW-based Mail
User Agent for Secure Electronic Mail Service for
HealthCare Users, Methods of Information in
Medicine,37,247-253,1998.
18) C.Wang, K.Ohe: A CORBA-based Object Frameworkwith Patient Identification Translation and Dynamic
Linking-Methods for Exchanging Patient Data,Method of information in Medicine,38(1),5665,1999.
19) D. Koide, K. Ohe, Dennis Ross-Degnan, S.Kaihara:
Computerized reminders to monitor liverfunction to
improve the use of etretinate. International Journalof Medical Informatics,57(2000),11-19, 2000.
20) E.Hanada, Y.Antoku, S.Tani, M.Kimura,
A.Hasegawa, S.Urano, K Ohe, Member, IEEE,
M.Ymaki, Y.Nose: Electromagnetic interference on
medical equipment by low-power mobile telecommunication systems. IEEE TRANSACTIONS ON ELECTROMAGNETIC COMPATIBILITY, 42(4),470-476,
2000.
21) K. Ohe, K.Miyo, Y.Onogi, K.Ueda, M.Takada,T.Chihara: Implications of a General data model for
implementing OODB/CORBA-based computerized
patient record system. Proceedings of MEDINFO2001, V.Patel etal.(Eds), Amsterdam:IOS
press,789,2001.
22) T.Kiuchi,K. Ohe,T.Sakurai: UMIN-Key information
infrastructure for the Japanese Medical Community.
Proceedings of MEDINFO2001, V.Patel etal.(Eds),
Amsterdam:IOS press,1359-1363,2001.
23) D.Koide,K. Ohe: Applying data mining to detectionof adverse drug reactions. Proceedings of MEDINFO2001, V.Patel etal.(Eds), Amsterdam:IOS
press,1421,2001.
24) S.Kataoka, K. Ohe, M.Mochizuki, S.Ueda: Developing
and integrating an adverse drug reaction reporting
system with the hospital information system. YAKUGAKU ZASSHI,122(1),113-116, 2002.
25) Y.Ohyama, K.Funao, E.Kawabe, D.Hayashi,
T.Yamazaki, T.Iga, D.Koide, K. Ohe, K.Kubota：
Calcium channel blockers and myocardial infarction:
A case-control study in a Japanese hospital. PHARMACOEPIDEMIOLOGY AND DRUG SAFETY, 2002,
11, 487-492, 2002.

156

Department of
Cardiovascular Medicine
Outline and Research Objectives
In 1998 the Department of Internal Medicine at University of Tokyo hospital was reorganized to the more functional units based on clinical specialties of diseased organs.
Cardiologists from 5 departments of Internal Medicine were unified to the Department of
Cardiovascular Medicine. We at quite an early phase introduced techniques of molecularbiology into the research field of cardiovascular medicine. As a result, we could report following important findings: 1) phenotypic modulation of cardiac or vascular specific proteins
occurs during cardiac hypertrophy or arteriosclerosis. 2) intracellular mechanisms on how
mechanical stress causes cardiac hypertrophy; 3) identification of novel genes expressed
during heart development; 4) development of animal models for cardiovascular diseases byusing genetic engineering techniques (i.e. knockout mice of endothelin-1 and KLF5 genes
provided new findings for the research of this field); 5) therapeutic angiogenesis for experimental myocardial infarction by growth factors was originally introduced in this department; 6) We are developing an electronic patient record system by which prognosis as wellas risk factors of cardiovascular disease can be statistically analyzed. From a research standpoint, our interests range throughout all fields of cardiovascular medicine ranging frommolecular biology to clinical research including genomics. Importantly, our research interests are aimed at making possible new diagnostics and treatment of cardiovascular diseases.
Areas of interest are as follows:
1) Transcriptional regulation in cardiovascular pathogenesis
2) Risk factor analysis of cardiovascular disease3) Pathogenic mechanisms of cardiac hypertrophy and heart failure4) Immunological basis of myocarditis and dilated cardiomyopathy5) MRI in cardiovascular diseases
6) Early diagnosis of ischemic heart disease using radionuclide testing7) Aerobic threshold and cardiac rehabilitation
8) Anti-arrhythmia therapy using atrial remodeling9) Mechanism of post-PTCA restenosis10) Molecular mechanisms of reperfusion injury11) Genetic polymorphisms in cardiovascular disease12) Differentiation of smooth muscle cells13) Cardiac development
14) Gene expression and regulation in cardiomyocytes15) Mouse genetic models of cardiovascular diseases and vascular development
16) Nitric oxide and endothelial function17) Gene therapy of heart failure using cardiac contractile proteins18) Clinical application of vasoactive substances

Faculties and Students
Professor and Chair
Lecturer

Ryozo Nagai, MD, PhD (1999~)
Yasunobu Hirata, MD, PhD

Minoru Ohno, MD, PhD
Yoshinori Seko, MD, PhD
Associate ...............................14
Postdoctoral Fellow ..............11
Graduate student ..................28
Research student.....................7
Secretary .................................4

Past Research and Major Accomplishments
1) Transcriptional regulation of expression ofgenes related to cardiovascular diseases
We have been focusing our research on the role oftranscriptional regulation of gene expression in the
cardiovascular system with a particular interest on
regulation of phenotypic modulation of smooth muscle and cardiac cells. In 1980s and early 1990s, Nagai
et al isolated and characterized three types of smooth157

muscle myosin heavy chains and found that they are
differentially expressed during vascular development.
Nagai et al furthermore isolated a DNA-binding factor,
Krüppel-like factor 5 (KLF5), as a transcription factorof the embryonic isoform of smooth muscle myosin
heavy chain gene (SMemb), whose expression is
induced in phenotypically modulated smooth muscle
cell and cardiac fibroblast. Recently, by developing
knockout mice of KLF5 gene, we have found that
KLF5 is an essential regulator of cardiovascularremodeling which occurs in response to various external stresses (Figure 1). We have further found that differential chemical modifications and protein-protein
interactions regulate this family of factors. We are currently investigating whether low molecular weightcompounds that inhibit functions of KLF5 are clinically applicable as a new therapeutic drug against cardiovascular diseases.
2) Differentiation of smooth muscle cells in vascular lesions
On the contrary to general assumption that neointima cells are derived from medial smooth muscle
cells, we found that bone marrow cells give rise to themajority of smooth muscle cells that contribute toarterial remodeling in models of post-angioplasty
restenosis, graft vasculopathy and hyperlipidemiainduced atherosclerosis. Notably, we found purified
hematopoietic stem cells differentiated into smoothmuscle cells in vitro and in vivo. We proposed thatamong blood cells there may be progenitors ofsmooth muscle cells, which attach to the injured
endothelia, differentiate into smooth muscle cells and158

proliferate, contributing thereby to neointima formation. Our results will also suggest a novel strategy toprevent vascular diseases, targeting mobilization (A),circulation (B), homing (C), differentiation (D) and proliferation (E) of putative smooth muscle progenitor
cells (Figure 2).
3) Establishment of clinical data management system and its practical application to genetic epidemiology in cardiovascular medicine
We have established a clinical data analysis systemon network, which enrolls more than 2000 patients
who underwent coronary angiography in our department. By using this system we can analyze the relationship between the genetic polymorphisms and theclinical of cardiovascular diseases such as coronary
artery diseases (CAD) and ischemic stroke. We
demonstrated that the alanine/valine (A/V) polymorphism in the gene of 5,10-methylenetetrahydrofolate
reductase (MTHFR) geneis a significant genetic risk
factor for CAD and ischemic stroke. This genetic variant is a more powerful predictor of atherosclerotic diseases, when the plasma folate levels are relatively low.
4) Development of mouse genetic models of cardiovascular diseases
Our research interests include the pathophysiological assessment of cardiovascular diseases by micegene engineering approach. We have generated
knockout and transgenic mice of adrenomedullin,
ADAMTS-1, endothelin-1 and KLF5. We analyzed therole of these genes and found that their expression isessential for normal growth of mice.

Figure 3 summarizes our strategy for clinical application of basic research on cardiovascular disease.

Current Research
1) Transcription factor such as Krüppel-like factors in pathogenic mechanisms.
Modulation of transcription factors may lead to a
new therapeutic strategy for cardiovascular disease.
Ongoing studies include using proteomics approaches
to understand the diverse protein-protein interactions
and regulation of these Krüppel-like factors. We havealready identified a novel repressor protein among
others. Another focus of research surrounds understanding the transcriptional regulation of these
Krüppel-like factors in the context of chromatin
which is necessary to understand how transcription
occurs in humans.
2) Bone marrow-derived vascular progenitor cells(1) Identification of bone marrow fraction contributing to atherosclerosis
To determine which fraction of bone marrow cells
contribute to the pathogenesis of vascular diseases,
we are reconstituting bone marrow of the lethally
irradiated recipient mice with the hematopoietic stem
cells from LacZ mouse and the mesenchymal stem
cells from GFP mouse. We are applying following vascular injury models to bone marrow-reconstitution
mice: mouse femoral injury model, heterotopic cardiac
transplantation model and atherosclerotic model.
(2) Characterization of bone marrow-derived
vascular progenitor cells
By using cell sorter, we are identifying which frac-tion of peripheral blood cells effectively differentiates
into smooth muscle cells. We are determining the cellsurface marker to isolate the putative bone marrowderived smooth muscle progenitor cells. Using bonemarrow transplantation mice, we are identifying theeffects of hyperlipidemia and drugs on mobilization,
homing, differentiation of the progenitor cells in various models of vascular injury described above3) Establishment of clinical data management system.
To overcome the difficulty of filing complicatedclinical parameters for genetic association studies, weare accumulating clinical data and constructing ouroriginal database system. For genetic analyses, written informed consent to participate was obtained
from all the in-patients in our department. DNA samples obtained from the participants have reachedapproximately one thousand. Using them, we analyzed over 50 genetic polymorphisms implicated to be
associated with atherosclerotic diseases. Among plenty of SNPs analyzed in our study, we showed thepolymorphisms in the MMP-1 and MMP-3 promoters
are associated with disease susceptibility to myocardial infarction. Also, our database system supports
several ongoing clinical epidemiological studies suchas J-CAD.
4) Genetic engineering for analysis of the role ofendogenous protein
We are analyzing the in vivo function of KLF5
using knockout mice. In response to external stress,

159

KLF5 knockout mice exhibited diminished levels ofarterial wall thickening, cardiac hypertrophy, interstitial fibrosis and that angiotensin II induced expression of KLF5, which in turn activated growth factor
expression. KLF5 thus appears to be a key element
linking external stress and cardiovascular remodeling.

Future Prospects
1) Dissection of the pathogenic mechanisms whereby
the Krüppel-like factors are involved in cardiovascular disease will allow us to understand thediverse mechanisms by which this family of factors are involved in transcriptional regulation. Bysuch, we envision that targeted drug design tomodulate these factors in this context will enable
us to propose a new therapeutic approach to cardiovascular disease (e.g. pin-point drug design).
2) Using our database system we have followed all
the cases periodically, thus we can get prospective
clinical data such as major cardiovascular eventsand responsiveness to treatment such as drugs andpercutaneous coronary intervention. In future, wewill be able to obtain several evidences from ourdatabase system and genetic analyses, resulting inclinical application of such evidences in prediction
or prevention of major cardiovascular diseases.
3) We have shown that KLF5 is a crucial determinantof the cellular response to cardiovascular injury,
playing a key role in mediating cardiovascularremodeling. We are now analyzing the effect ofcompounds, which could modulate KLF5 function
to control remodeling in atherosclerosis and heart
failure.
4) Our studies will show that bone marrow cells
including hematopoietic stem cells have the potential to give rise to vascular progenitor cells thathome in the damaged vessels, differentiate into
smooth muscle cells, and proliferate thereby contributing to vascular remodeling. We expect thatatherogenic factors such as hyperlipidemia, inflammation, cytokines, and smoking will facilitate thekinetics of smooth muscle progenitors. Our findings will provide the basis for the development of
new diagnostic strategies to predict atherosclerosis
by quantifying the circulating progenitors.

Research Grants
Ryozo Nagai (1997-2001)
Grant from the Organization for Pharmaceutical
Safety and ResearchRole of Klotho gene and its clinical application in cardiovascular disease

160

Ryozo Nagai (1999-2001)
Grant from the Ministry of Health, Labour andWelfareResearch on elucidation of the molecular mechanisms of specific diseases
Ryozo Nagai (2000-2002)
Grant from the Ministry of Education, Culture,
Sports, Science and Technology
Development of a new prognostic evaluation systemand treatment for patients with cardiovascular disease based on gene polymorphisms
Ryozo Nagai (2000-2002)
Grant from the Ministry of Health, Labor andWelfareResearch on development of angiogenesis and vascular protection treatment
Ryozo Nagai (2002-2006)
Grant from the Ministry of Education, Culture,
Sports, Science and Technology
Molecular mechanism and organ remodeling: genetranscription and cell-cell interaction in mesenchymal cells

Select Publications
Diversity of smooth muscle myosin heavy chain
isoforms
1. Nagai R, Larson DM, Periasamy M. Characterizationof a mammalian smooth muscle myosin heavy chain
cDNA and its expression in various smooth muscle
types. Proc Natl Acad Sci USA 85: 1047-1051, 1988.
2. Nagai R, Kuro-o M, Babij P, Periasamy M.
Identification of two types of smooth muscle myosin
heavy chain isoforms by cDNA cloning andimmunoblot analysis. J Biol Chem 264: 9734-9737,
1989.
3. Kuro-o M, Nagai R, Tsuchimochi H, Katoh H, Yazaki
Y, Ohkubo A, Takaku F. Developmentally regulated
expression of vascular smooth muscle myosin heavy
chain isoforms. J Biol Chem 264: 18272-18275, 1989.
4. Kuro-o M, Nagai R, Tsuchimochi H, Katoh H, Tsai RC, Yazaki Y, Ohkubo A, Takaku F. cDNA cloning of amyosin heavy chain isoform in embryonic smoothmuscles and its expression during vascular development and in arteriosclerosis. J Biol Chem 266: 37683773, 1991.
5. Kim H-S, Aikawa M, Kimura K, Kuro-o M, Nakahara
K, Suzuki T, Katoh H, Okamoto E, Yazaki Y, Nagai R.
Ductus arteriosus: advanced differentiation ofsmooth muscle cells demonstrated by myosin heavy
chain isoform expression in rabbits. Circulation 88:
1804-1810, 1993.
6. Aikawa M, Nalla Sivam P, Kuro-o M, Kimura K,
Nakahara K, Takewaki S, Yamaguchi H, Yazaki Y,
Periasamy M, Nagai R. Human smooth muscle
myosin heavy chain isoforms as molecular markers
for vascular development and atherosclerosis. Circ
Res 73: 1000-1012, 1993.

7. Suzuki T, H-S Kim, Kurabayashi M, Hamada H, Fujii
H, Aikawa M, Watanabe M, Yazaki Y, Nagai R
Preferential differentiation of P19 mouse embryonalcarcinoma cells into smooth muscle cells: use ofretinoic acid and antisense against the central nervous system-specific POU transcription factor Brn-2.
Circ Res 78:395-404,1996.

tribute to atherosclerosis. Nature Med. 7: 382-383,
2001.
9. Sata M, Saiura A, Kunisato A, Tojo A, Okada S,
Tokuhisa T, Hirai H, Makuuchi M, Hirata Y, Nagai R.
Hematopoietic stem cells differentiate into vascular
cells that participate in the pathogenesis of atherosclerosis. Nature Med. 8: 403-409, 2002.

Regulation of smooth muscle cell differentiationand cardiovascular remodeling
1. Watanabe M, Sakomura Y, Kurabayashi M, Manabe
I, Aikawa M, Kuro-o M, Suzuki T, Yazaki Y, Nagai R
Structure and characterization of the 5’-Flanking
Region of the mouse smooth muscle myosin heavy
chain (SM1/2) gene. Circ Res 78: 978-989, 1996.
2. Manabe I, Kurabayashi M, Shimomura Y, Kuro-o M,
Watanabe N, Watanabe M, Aikawa M, Suzuki T,
Yazaki Y, Nagai R Isolation of the embryonic form ofsmooth muscle myosin heavy chain (SMemb/NMHCB) gene and characterization of its 5’-flanking region.
Biochem Biophys Res Commun 239:598-605, 1997.
3. Watanabe N, Kurabayashi M, Shimomura Y, KawaiKowase K, Hoshino Y, Manabe I, Watanabe M,
Aikawa M, Kuro-o M, Suzuki T, Yazaki Y, Nagai R
BTEB2, a Krppel-like transcription factor, regulates
expression of the SMemb/Nonmuscle myosin heavy
chain B (SMemb/NMHC-B) gene Circ Res 85: 182191, 1999.
4. Kawai-Kowase K, Kurabayashi M, Hoshino Y,
Ohyama Y, Nagai R. Transcriptional activation of thezinc finger transcription factor BTEB2 gene by Egr-1
through mitogen-activated protein kinase pathways
in vascular smooth muscle cells Circ Res 85: 787-795,
1999.
5. Hoshino Y, Kurabayashi M, Kanda T, Hasegawa A,
Sakamoto H, Okamoto Ei, Kowase K, Watanabe N,
Manabe I, Suzuki T, Nakano A, Takase Si, Wilcox
JN, Nagai R. Regulated expression of the BTEB2 transcription factor in vascular smooth muscle cells :analysis of developmental and pathological expression profiles shows implications as a predictive factor for restenosis. Circulation 102: 2528-2534, 2000.
6. Sekiguchi K, Kurabayashi M, Oyama Y, Aihara Y,
Tanaka T, Sakamoto H, Hoshino Y, Kanda T,
Yokoyama T, Shimomura Y, Iijima H, Ohyama Y,
Nagai R. Homeobox protein hex induces SMemb/
Nonmuscle myosin heavy chain-B gene expression
through the cAMP-responsive element. Circ Res 88:
52-28, 2001.
7. Shindo T, Manabe I, Fukushima Y, Tobe K, Aizawa
K, Miyamoto S, Kawai-Kowase K, Moriyama N, Imai
Y, Kawakami H, Nishimatsu H, Ishikawa T, Suzuki
T, Morita H, Maemura K, Sata M, Hirata Y, Komukai
M, Kagechika H, Kadowaki T, Kurabayashi M, Nagai
R. Krppel-like zinc-finger transcription factor
KLF5/BTEB2 is a target for angiotensin II signaling
and an essential regulator of cardiovascular remodeling. Nature Med. 8:856-873, 2002.
8. Saiura A, Sata M, Hirata Y, Nagai R, Makuuchi M.
Circulating smooth muscle progenitor cells con-Identification and function of anti-aging geneklotho1. Kuro-o M, Matsumura Y, Aizawa H, Kawaguchi H,
Suga T, Utsugi T, Ohyama Y, Kurabayashi M,
Kaname T, Kume E, Iwasaki H, Iida A, Shiraki-Iida T,
Nishikawa S, Nagai R, Nabeshima Y. Mutation of themouse Klotho gene leads to a syndrome resembling
ageing. Nature 390: 45-51, 1997.
2. Saito Y, Yamagishi T, Nakamura T, Ohyama Y,
Aizawa H, Suga T, Matsumura Y, Masuda H,
Kurabayashi M, Kuro-o M, Nabeshima Y, Nagai R.
Klotho protein protects against endothelial dysfunction. Biochem Biophys Res Commun 248: 324-329,
1998.
3. Aizawa H, Saito Y, Nakamura T, Inoue M, Imanari T,
Ohyama Y, Matsumura Y, Masuda H, Oba S, Mise N,
Kimura K, Hasegawa A, Kurabayashi M, Kuro-o M,
Nabeshima Y, Nagai R. Downregulation of the klotho
gene in the kidney under sustained circulatory stressin rats. Biochem Biophys Res Commun 249: 865-871,
1998.
4. Suga T, Kurabayashi M, Sando Y, Ohyama Y, Maeno
T, Maeno Y, Aizawa H, Matsumura Y, Kuwaki T,
Kuro-o M, Nabeshima Y, Nagai R. Disruption of theklotho gene causes pulmonary emphysema in mice:
defect in maintenance of pulmonary integrity during
postnatal life Am J Respir Cell Mol Biol 22: 26-33,
2000.
5. Nagai R, Saito Y, Ohyama Y, Aizawa H, Suga T,
Nakamura T, Kurabayashi M, Kuro-o M Endothelial
dysfunction in the klotho mouse and downregulation of klotho gene expression in various animalmodels of vascular and metabolic diseases. Cell Mol
Life Sci 57: 738-746, 2000.
6. Saito Y, Nakamura T, Ohyama Y, Suzuki T, Iida A,
Shiraki-Iida T, Kuro-o M, Nabeshima Y, Kurabayashi
M, Nagai R. In vivo klotho gene delivery protects
against endothelial dysfunction in multiple risk factor syndrome. Biochem Biophys Res Commun
276:767-772, 2000.
7. Mitani H, Ishizaka N, Aizawa T, Ohno M, Usui S,
Suzuki T, Amaki T, Mori I, Nakamura Y, Sato M,
Nangaku M, Hirata Y, Nagai R. In vivo klotho genetransfer ameliorates angiotensin II-induced renal
damage. Hypertension. 39:838-843, 2002.
Intracellular signaling and transcription factors ofcardiac hypertrophy
1. Kojima M, Shiojima I, Yamazaki T, Komuro I, Nagai
R, Yazaki Y. Angiotensin II receptor antagonist
TCV116 regresses hypertensive left ventricular
hypertrophy in vivo and inhibits intracellular signal-161

ing pathway of stretch-mediated cardiomyocyte
hypertrophy in vitro. Circulation 89: 2204-2211,
1994
2. Shimoyama M, Hayashi D, Takimoto E, Zou Y, Oka
T, Uozumi H, Kudoh S, Shibasaki F, Yazaki Y, Nagai
R, Komuro I. Calcineurin plays a critical role in pressure overload-induced cardiac hypertrophy.
Circulation. 100:2449-2454, 1999.
3. Zhu W, Zou Y, Aikawa R, Harada K, Kudoh S,
Uozumi H, Hayashi D, Gu Y, Yamazaki T, Nagai R,
Yazaki Y, Komuro I. MAPK superfamily plays animportant role in daunomycin-induced apoptosis ofcardiac myocytes. Circulation. 100:2100-2107,1999.
4. Aikawa R, Nawano M, Gu Y, Katagiri H, Asano T,
Zhu W, Nagai R, Komuro I. Insulin prevents cardiomyocytes from oxidative stress-induced apoptosis
through activation of PI3 kinase/Akt. Circulation.
102: 2873-2879, 2000.
5. Hiroi Y, Kudoh S, Monzen K, Ikeda Y, Yazaki Y,
Nagai R, Komuro I. Tbx5 associates with Nkx2-5 andsynergistically promotes cardiomyocyte differentiation. Nature Genet. 28:276-280, 2001.
6. Hosoda T, Monzen K, Hiroi Y, Oka T, Takimoto E,
Yazaki Y, Nagai R, Komuro I. A novel myocyte-specific gene Midori promotes the differentiation of
P19CL6 cells into cardiomyocytes. J Biol Chem.
276:35978-35989, 2001.
7. Uozumi H, Hiroi Y, Zou Y, Takimoto E, Toko H, Niu
P, Shimoyama M, Yazaki Y, Nagai R, Komuro I.
gp130 plays a critical role in pressure overloadinduced cardiac hypertrophy. J Biol Chem. 276:
23115-23119, 2001.
8. Zou Y, Hiroi Y, Uozumi H, Takimoto E, Toko H, Zhu
W, Kudoh S, Mizukami M, Shimoyama M, Shibasaki
F, Nagai R, Yazaki Y, Komuro I. Calcineurin plays acritical role in the development of pressure overloadinduced cardiac hypertrophy. Circulation. 104:97101, 2001.
9. Zou Y, Hiroi Y, Uozumi H, Takimoto E, Toko H, Zhu
W, Kudoh S, Mizukami M, Shimoyama M, Shibasaki
F, Nagai R, Yazaki Y, Komuro I. Calcineurin plays acritical role in the development of pressure overloadinduced cardiac hypertrophy. Circulation. 104:97101, 2001.
10. Takimoto E, Yao A, Toko H, Takano H, Shimoyama
M, Sonoda M, Wakimoto K, Takahashi T, Akazawa
H, Mizukami M, Nagai T, Nagai R, Komuro I. Sodium
calcium exchanger plays a key role in alteration ofcardiac function in response to pressure overload.
FASEB J 16:373-378, 2002.

Cardiovascular pathophysiology
1. Kurihara Y, Kurihara H, Suzuki H, Kodama T,
Maemura K, Nagai R, Oda H, Kuwaki T, Cao WH,
Kamada N, Jishage K, Ouchi Y, Azume S, Toyoda Y,
Ishikawa T, Kumada M, and Yazaki Y. Elevatedblood pressure and craniofacial abnormalities in mice
deficient in endothelin-1. Nature 368: 703-710, 1994.
2. Kuro-o M, Hanaoka K, Hiroi Y, Noguchi T, Fujimori
Y, Takewaki S, Hayasaka M, Katoh H, Miyagishi A,

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3.

4.

5.

6.

7.

8.

Nagai R, Yazaki Y, Nabeshima Y. Salt-sensitinve
hypertension in transgenic mice overexpressing sodium-proton exchanger. Circ Res 76: 148-153, 1995.
Shindo T, Kurihara H, Kuno K, Yokoyama H, Wada
T, Kurihara Y, Imai T, Wang Y, Ogata M, Nishimatsu
H, Moriyama N, Oh-hashi Y, Morita H, Ishikawa T,
Nagai R, Yazaki Y, Matsushima K. ADAMTS-1: ametalloproteinase-disintegrin essential for normal
growth, fertility, and organ morphology and function. J Clin Invest. 105:1345-1352, 2000.
Shindo T, Kurihara H, Maemura K, Kurihara Y,
Kuwaki T, Izumida T, Minamino N, Ju KH, Morita H,
Oh-hashi Y, Kumada M, Kangawa K, Nagai R, Yazaki
Y. Hypotension and resistance to lipopolysaccharideinduced shock in transgenic mice overexpressing
adrenomedullin in their vasculature. Circulation.
101:2309-2316, 2000.
Shindo T, Kurihara Y, Nishimatsu H, Moriyama N,
Kakoki M, Wang Y, Imai Y, Ebihara A, Kuwaki T, Ju
KH, Minamino N, Kangawa K, Ishikawa T, Fukuda
M, Akimoto Y, Kawakami H, Imai T, Morita H,
Yazaki Y, Nagai R, Hirata Y, Kurihara H. Vascularabnormalities and elevated blood pressure in micelacking adrenomedullin gene. Circulation. 104:19641971, 2001.
Morita H, Kurihara H, Yoshida S, Saito Y, Shindo T,
Oh-Hashi Y, Kurihara Y, Yazaki Y, Nagai R. Dietinduced hyperhomocysteinemia exacerbates neointima formation in rat carotid arteries after balloon
injury. Circulation. 103:133-139, 2001
Yasuda SI, Sugiura S, Kobayakawa N, Fujita H,
Yamashita H, Katoh K, Saeki Y, Kaneko H, Suda Y,
Nagai R, Sugi H. A novel method to study contraction characteristics of a single cardiac myocyte using
carbon fibers. Am J Physiol Heart Circ Physiol.
281:H1442-6, 2001.
Ishizaka N, Aizawa T, Ohno M, Usui Si S, Mori I,
Tang SS, Ingelfinger JR, Kimura S, Nagai R.
Regulation and localization of HSP70 and HSP25 in
the kidney of rats undergoing long-term administration of angiotensin II. Hypertension. 39:122-128,
2002.

NO and intracellular signaling pathways
1. Nagata D, Suzuki E, Nishimatsu H, Yoshizumi M,
Mano T, Walsh K, Sata M, Kakoki M, Goto A, Omata
M, Hirata Y. Cyclin A downregulation and p21(cip1)
upregulation correlate with GATA- 6-induced growtharrest in glomerular mesangial cells. Circ Res. 87:
699-704, 2000.
2. Sata M, Kakoki M, Nagata D, Nishimatsu H, Suzuki
E, Aoyagi T, Sugiura S, Kojima H, Nagano T,
Kangawa K, Matsuo H, Omata M, Nagai R, Hirata Y.
Adrenomedullin and nitric oxide inhibit humanendothelial cell apoptosis via a cyclic GMP-independent mechanism. Hypertension. 36: 83-88, 2000.
3. Suzuki E, Nagata D, Yoshizumi M, Kakoki M, Goto
A, Omata M, Hirata Y. Reentry into the cell cycle ofcontact-inhibited vascular endothelial cells by aphosphatase inhibitor. Possible involvement of

4.

5.

6.

7.

extracellular signal-regulated kinase and phosphatidylinositol 3-kinase. J Biol Chem. 275: 36373644, 2000
Nagata D, Suzuki E, Nishimatsu H, Satonaka H, Goto
A, Omata M, Hirata Y. Transcriptional activation of
the cyclin D1 gene is mediated by multiple cis-elements, including SP1 sites and a cAMP-responsive
element in vascular endothelial cells. J Biol Chem.
276: 662-669, 2001.
Nishimatsu H, Suzuki E, Nagata D, Moriyama N,
Satonaka H, Walsh K, Sata M, Kangawa K, Matsuo
H, Goto A, Kitamura T, Hirata Y. Adrenomedullin
induces endothelium-dependent vasorelaxation via
the phosphatidylinositol 3-kinase/Akt-dependent
pathway in rat aorta.Circ Res. 89: 63-70, 2001.
Nishimatsu H, Hirata Y, Shindo T, Kurihara H,
Kakoki M, Nagata D, Hayakawa H, Satonaka H, Sata
M, Tojo A, Suzuki E, Kangawa K, Matsuo H,
Kitamura T, Nagai R. Role of endogenous
adrenomedullin in the regulation of vascular tone
and ischemic renal injury: studies ontransgenic/knockout mice of adrenomedullin gene.
Circ Res. 90: 657-663, 2002.
Suzuki N, Kojima H, Urano Y, Kikuchi K, Hirata Y,
Nagano T. Orthogonality of calcium concentration
and ability of 4,5- diaminofluorescein to detect NO. J
Biol Chem 277:47-49, 2002.

Nagai R. A novel biochemical diagnostic method for
aortic dissection - the results of a prospective study
using an immunoassay of smooth muscle myosin
heavy chain. Circulation 93:1244-1249, 1996
2. Yokoyama I, Yonekura K, Ohtake T, Yang W, Shin
WS, Yamada N, Ohtomo K, Nagai R. Coronary
microangiopathy in type 2 diabetic patients: relation
to glycemic control, sex, and microvascular angina
rather than to coronary artery disease. J Nucl Med
41: 978-985, 2000
3. Iwasaki T, Iwasaki T, Aihara Y, Kanda T, Oriuchi N,
Endo K, Katoh H, Suzuki T, Nagai R.
Immunoscintigraphy of aortic dissection with
99mTc-labeled murine anti-smooth muscle myosin
monoclonal antibody in rats. J Nucl Med 42: 130137, 2001
4. Yokoyama I, Yonekura K, Moritan T, Tateno M,
Momose T, Ohtomo K, Inoue Y, Nagai R.
Troglitazone improves whole-body insulin resistanceand skeletal muscle glucose use in type II diabetic
patients. J Nucl Med. 42:1005-1010, 2001.

Genetics of cardiovascular disease
1. Tanaka T, Nagai R, Tomoike H, Takata S, Yano K,
Yabuta K, Haneda N, Nakano O, Shibata A,
Sawayama T, Kasai H, Yazaki Y, Nakamura Y. Fournovel KVLQT1 and four novel HERG mutations infamilial long QT syndrome. Circulation 95: 565-567,
1997.
2. Nakajima T, Furukawa T, Tanaka T, Katayama Y,
Nagai R, Nakamura Y, Hiraoka M. Novel mechanismof HERG current suppression in LQT2 shift in voltage dependence of HERG inactivation. Circ Res
83:415-422, 1998.
3. Itoh T, Tanaka T, Nagai R, Kamiya T, Sawayama T,
Nakayama T, Tomoike H, Sakurada H, Yazaki Y,
Nakamura Y. Genomic organization and mutational
analysis of HERG, a gene responsible for familial long
QT syndrome. Hum Genet 102: 435-439, 1998
4. Itoh T, Tanaka T, Nagai R, Kikuchi K, Ogawa S,
Okada S, Yamagata S, Yano K, Yazaki Y, Nakamura
Y. Genomic organization and mutational analysis of
KVLQT1, a gene responsible for familial long QT
syndrome. Hum Genet 103: 290-294, 1998
5. Iwasa H, Itoh T, Nagai R, Nakamura Y, Tanaka T.
Twenty single nucleotide polymorphisms (SNPs) andtheir allelic frequencies in four genes that are responsible for familial long QT syndrome in the Japanese
population. J Hum Genet 45: 182-183, 2000
Diagnosis and imaging of cardiovascular disease
1. Suzuki T, Katoh H, Watanabe M, Kurabayashi M,
Hiramori K, Hori S, Nobuyoshi M, Tanaka H,
Kodama K, Sato H, Suzuki S, Tsuchio Y, Yazaki Y,

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Department of
Gastroenterology
Department Objectives
The goal of our department is to accurately diagnose and give the best available treatment to patients, and develop better modalities for the future.
To attain the goal, we perform clinical and basic research.

OutlineIn 1998, our current Department of Gastroenterology was established through the reorganization of the Divisions of Internal Medicine, and is in charge of busy clinical practices;
5,000 outpatients a month and 1,200 new admissions a year. The most frequent cause ofadmission is hepatocellular carcinoma, 500 a year, followed by gastrointestinal tract disorders 400, pancreatobiliary 180, and others 120.
Outline-Hepatology
Unlike other countries, the majority (more than 90%) of our patients with liver disease
are due to hepatitis virus infection. Therefore, better understanding of virus replication andpathogenesis may eventually lead to the eradication of the virus and the cure of diseases.
We have developed unique ablation treatment for hepatocellular carcinoma (HCC), forwhich many patients have visited our department (Fig. 1); we have had 1,600 cases, probably the largest series in the world as a single institute experience.
Outline -Gastroenterology
Gastric cancer is the commonest cause of death in Japan. Similar to HCC, there is now
enough evidence that bacterial infection, Helicobacter pylori, induces or is at least strongly
related to cancer development. Along with clinical expertise like en bloc resection of cancer(Fig. 2), better understanding of gastric injury due the infection could reduce the most common cause of death in Japan.
Recent increase in the number of colonic cancer is so noticeable. We found that this ismainly due to increase in right sided colon (cecum and ascending colon) cancer in elderly
patients. The better understanding of right sided as well as left sided colonic cancers may
prevent further increase in the colonic neoplasm in a country where people enjoy unprecedented length of life.
Outline -Pancreatobiliary
Pancreatic cancer with incredibly poor prognosis (average survival of 200 days) is insidiously increasing in Japan. There is not even a clue for high risk group. We need, though difficult, to find out high risk groups and better treatment.
Fig 1. Percutaneous tumor ablation.
(A) During treatment: fine needle was inserted into the tumor.
(B) After treatment: the tumor was completely ablated

Fig 2. Endoscopic tumor resection.
(A) After injection of saline, the tumor was cut around.
(B) en bloc resection: the tumor was completely removed.

164

Faculties and Students
Professor and Chair

Masao Omata, M.D., Ph.D. (from1992)
Lecturer
Takao Kawabe, M.D., Shin Onishi,
M.D.
Associate ...............................19
Postgraduate Fellow ............33
Graduate Student..................23
Research Studen .....................7
Secretary .................................2

Past Research and Major Accomplishments
1. Acute and Chronic Liver Injury due to HBV(Hepatitis B Virus)
The most severe form of acute liver injury is fulminant hepatitis of which 70% is fatal.
However, the underlying mechanisms of severe
injury have not been clear.
We elucidated for the first time the fulminant hepatitis was induced by HBV mutant form (N Engl J Med
1991;324:1699-1704).
Furthermore, this mutant carries specific aminoacid changes in nucleo-capsid region, a possible targetof lymphocyte attack (J Clin Invest 1993;91:1206 1213).
These mutations play a critical role not only inacute but also chronic liver disease (J Clin Invest 1992
;89:332-338, Gastroenterology 1993;104:263-271).
By these understandings, we explored how totreat these wild and mutant strains.
We set up in vitro system to maintain these wild
and mutant virus replication and identified extremely
effective drugs (J Clin Invest1999;103:1635-1640, J
Clin Invest 2001;107:449-455).
These drugs are now on clinical trials to prevent
fulminant hepatitis and even the decompensated cirrhosis to save patients life.
2. Liver Injury due to HCV (Hepatitis C Virus)
In contrast to HBV, HCV infection is mild but
insidious and long-lasting disease to carcinoma.
All of these start with very high chronicity rate(80%) of acute hepatitis. We elucidated for the firsttime that this is nearly 100% preventable by anti-viraltreatment(Lancet1991;338:914-915,
Gastroenterology 1994;107:805-811). Since HCV vaccine has not been developed, the above mentioned isthe best available procedure to prevent chronicity. In
fact, a study from Germany recently reported identicalresults (N Engl J Med 2001;345:1452-1457). However,there are stil1 2 million carriers who already havechronic infection. In fact, over 80% of patients with
HCC are due to HCV infection in Japan. To prevent

HCC, we first revealed natural course of chronic hepatitis to HCC ; this is by step-wise progression of hepatic fibrosis (Ann Intern Med 2000;132:517-524). And
eradication of virus by interferon could reduce theincidence of HCC (Ann Intern Med 1999;131:174-181)
with resolution of hepatic fibrosis even in cirrhosis,
previously thought irreversible (Ann Intern Med
2000;132:517-524). Furthermore, it was recently
revealed that overall mortality (not just HCC) was
reduced in treatment cohort (Gastroenterology
2002;123: 483-491). By these studies, we now know
how to prevent, but question is how to treat patients
who already have the cancer. Probably our department is the largest referral center of HCC (500 cases ayear), some are from outside Japan, seeking our medical tumor ablation treatment (Fig. 1).
We recently finished our 7-year prospective randomized trial in that we combined radical cure of thecancer nodules by ablation and eradication of HCV byinterferon. The 7-year survival of these patientsreached 75% (Ann Intern Med 2002; in press). This iscompatible or somewhat better than those reported inliver transplantation. When I graduated medical
school 30 years ago, liver disease was the one impossible to treat, but now able to envision to how to cureall.
3. Gastric Injury
Gastric cancer is number one cause of death inJapan. More than 80% is supposed to be due to H.
pylori infection; 90% eradication of the bacteria is now
possible by proper treatment. Thus, we decided toelucidate underlying mechanisms from infection to
the cancer, by in vitro and in vivo studies. First, we
tried to identify responsible molecules for inflammation.
In vitro
We revealed H. pylori induces strong
NF-kB signals, and these are by molecules in Cag
pathogenicity island, 40kb segment of 1.5 million
nucleotides (Gastroenterology 2000;119:97-108, J Biol
Chem 2001;276:44856-44864).
In vivo
To extrapolate this to in vivo system,
we constructed isogenic mutant (knock out bacteria),
lacking Cag gene molecules, and infected it to
Mongolian gerbils (unique animal model). It turned
out among Cag island, Cag E gene is the one responsible for strong induction of inflammation, whereas Vac
A, claimed to be inflammation inducing molecule, isnot (J Exp Med 2000;192:1601-1609). These CagE
molecule of H. pylori gene is generally present inJapanese isolates, whereas
it is often absent inEuropean (Gut 1998;42:338-343). These may explain
the difference of incidence of the gastric cancer
among different countries.
Recently, Kawakami and Tateishi further elucidated that IkBα, a key molecule in NF-kB signal cascade,165was regulated by NEDD8 pathway activating ubiquitin-proteasome degradation system (EMBO J
2001;20:4003-4012, J Cell Biol 2001;155:571-579).
Although the NF-kB signaling is extremely important as an inflammation inducing pathway, mechanism of direct effect on cell proliferation by H. pylori
is not known. Hirata recently elucidated that CagA
protein strongly activates SRE (Serum Response
Element) which might be operating in the formationof polyp and even MALT lymphoma(Gastroenterology 2002; in press).
Our search for effector protein to induce signalingcascades has now began (J Exp Med 2000;191:593602); it may help to eventually elucidate the molecularmechanism of gastric cancer.
4. Colon Cancer
Colonic cancer is catching up with the leading gastric cancer. Shift from Japanese to “Westernized” food
may be responsible for this increase. However, we
believe there is another reason. We have noticed rightsided (cecum and ascending) colon cancer is increasing among patients over age 70 (60% vs 20% in ages
over 70 and under 40, respectively) (Gastrointest
Endosc 2002;55:548-551). We found the majority of
the right sided colon cancer carried microsatellite
instability (Cancer Res 1996;56:5620-5623). Now inJapan, average life expectancy for female and male is
85 and 78, respectively, longest ever.
We have initiated genome-wild screening of thegenes having dinucleotide repeat or poly-A stretches,targets of repair gene errors.
5. Pancreatic Cancer
Once pancreatic cancer developed, median survival
was only 200 days. Although we tried to employ K-ras
gene mutation, which is found in 90% of the cancer,
for early diagnosis (Gastroenterology 1991;100:233238, 1996;110:227-231), it has been far from giving
benefit to patients.
We have struggled to find better way of treatment
and finally decided to apply allogenic stem cell transplantation by collaboration with Department of
Hematology and Oncology. So far 5 patients enrolled
and results are pending.

Current Research and Clinical Activities
1. Eradication of HBVAs I wrote, the ultimate goal of HBV treatment is
not mere suppression which is currently available, but
eradication of the virus (Treatment of chronic hepatitis B infection -Editorial N Engl J Med 1998 ; 339:114115). To attain this goal we are undertaking the following :166

HBV has two replication pathways, episomal (NEngl J Med 1986;315:1187-1192) and integrated form
into human genome which presumably resist thetreatment. We are now intensively focusing our study
on the integrated form by use of post-sequencing
information on human genome.
2. Eradication of HCV
We could eradicate the virus in more than 90% of
acute hepatitis cases as was shown by us (Lancet
1991;338:914-915). However, only 30% in chronic
hepatitis. To increase treatment efficacy, we now have
Replicon system which allows us to grow treatmentresistant strains and to test drug candidates.
3. Liver Injury Mechanisms
Sequencing of extreme small amount of peptides isnow feasible by high quality mass spectrometry. We
are now analyzing HLA-related wild and mutants
virus-derived peptides from human liver. This may
identify the target of lymphocyte attack.
4. Prevention and Cure of HCC
We have treated 1,600 cases of HCC so far. We
have shown that treatment of virus infection could
prevent HCC. However, advanced HCC is still difficult
to treat. We found that Vitamin K could efficiently
prevent portal tumor invasion, once developed 100%
fatal in one year. Along with other modalities, wemight expect long-term survival in the majority of thepatients with HCC.
5. Gastric Injury Mechanisms
Discovery of H. pylori protein in host cell, phosphorylated CagA protein (J Exp Med 2000;191:593602), has prompted us to further search the effector
protein, which could trigger signaling cascades. We
have now several candidates.
6. Right-sided Colon Carcinogenesis
As I mentioned, repair-gene system seems deeply
involved in the right-sided colon cancer, a prototype
is, of course, HNPCC (Hereditary Nonpolyposis Colon
Cancer). The understanding may help for people inthe country where longest life-expectancy expected.
7. Pancreatic Cancer
We have initiated a large scale genome-wide SNP(Single Nucleotide Polymorphism) analysis to findhigh risk group.

Others Activites
In 5 years, we presented 121 papers in DDW(Digestive Disease Week, USA), the largest gastroenterology congress.

In 5 years, we published 320 papers in peer review
journal.
In 5 years, 52 invited lectures were given in international meetings by one of us (M Omata).

Future Prospective
Education for medical students and for young
physicians is the far most important mission of ourdepartment for the future. Our department have tocover multiple organs with diverse diseases. Thus, weare trying not to restrict their interest to one specialized field, but rather to introduce general technology
and ideas. It will definitely become more “borderless”
in post-genome era.

Research Grants
The Organization for Pharmaceutical Safety
Research 1997-2001 (Total ￥346,000,000.)
Comprehensive 10-year Strategy for Cancer
Control from Ministry of Health and Welfare, Health
Sciences Research Grants 1994-1999 (Total￥100,600,000.)
Medical Frontier Strategy Research from Ministryof Health Labor and Welfare, Health Sciences
Research Grants 2001-2003 (2001 / ￥48,000,000.)
Grants-in-aid for Scientific Research on Priority
Area from the Ministry of Education, Culture, Sports,
Science and Technology of Japan 2000-2003 (Total￥71,800,000.)
Grants-in-aid for Scientific Research (A) from the
Ministry of Education, Culture, Sports, Science andTechnology of Japan 2001-2004 (Total ￥40,300,000.)

Select Publications (50)
Hepatology
1. Omata M, Ehata T, Yokosuka O, Hosoda K, Ohto M.
Mutations in the precore region of hepatitis B virus
DNA in patients with fulminant and severe hepatitis.
N Engl J Med 324 : 1699-1704, 1991.
2. Omata M, Yokosuka O, Takano S, Kato N, Hosoda K,
Imazeki F, Tada M, Ito Y, Ohto M. Resolution of
acute hepatitis C after therapy with natural beta
interferon. Lancet 338 : 914-915, 1991.
3. Ehata T, Omata M, Yokosuka O, Hosoda K, Ohto M.
Variations in codon 84-101 in the core nucleotide
sequence correlate with hepatocellular injury inchronic hepatitis B virus infection. J Clin Invest 89 :332-338, 1992.
4. Hosoda K, Omata M, Yokosuka O, Kato N, Ohto M.
Nucleotide sequences of 5- 1-1 of hepatitis C virus inpatients with chronic liver disease. Gastroenterology
102 :1039-1043, 1992.

5. Chien DY, Choo QL, Tabrizi A, Kuo C, McFarland J,
Berger K, Lee C, Shuster JR, Nguyen T, Moyer DL,
Tong M, Furuta S, Omata M, Tegtmeier G, Alter H,
Schiff E, Jeffers L, Houghton M, Kuo G. Diagnosis ofhepatitis C virus (HCV) infection using an immunodominant chimeric polyprotein to capture circulating antibodies : reevaluation of the role of HCV inliver disease. Proc Natl Acad Sci USA 89 : 1001110015, 1992.
6. Ehata T, Omata M, Chuang WL, Yokosuka O, Ito Y,
Hosoda K, Ohto M. Mutations in core nucleotide
sequence of hepatitis B virus correlate with fulminant and severe hepatitis. J Clin Invest 91 : 12061213, 1993.
7. Chuang WL, Omata M, Ehata T, Yokosuka O, Ito Y,
Imazeki F, Lu SN, Chang WY, Ohto M. Precore mutations and core clustering mutations in chronic hepatitis B virus infection. Gastroenterology 104 : 263271, 1993.
8. Takano S, Satomura Y, Omata M, Japan Acute
Hepatitis Cooperative Study Group. Effects of interferon beta on Non-A, Non-B acute hepatitis: A
prospective, randomized, controlled-dose study.
Gastroenterology 107 : 805-811, 1994.
9. Takano S, Yokosuka O, Imazeki F, Tagawa M, Omata
M. Incidence of hepatocellular carcinoma in chronic
hepatitis B and C : A prospective study of 251
patients. Hepatology 21 : 650-655, 1995.
10. Matsumoto A, Ogura K, Hirata Y, Kakoki M,
Watanabe F, Takenaka K, Shiratori Y, Momomura S,
Omata M. Increased nitric oxide in the exhaled air ofpatients with decompensated liver cirrhosis. Ann
Intern Med 123 : 110-113, 1995.
11. Shiratori Y, Shiina S, Imamura M, Kato N, Kanai F,
Okudaira T, Teratani T, Tohgo G, Toda N, Ohashi M,
Ogura K, Niwa Y, Kawabe T, Omata M.
Characteristic difference of hepatocellular carcinoma
between hepatitis-B and -C viral infection in Japan.
Hepatology 22 : 1027-1033, 1995.
12. Kanai F, Lan KH., Shiratori Y, Tanaka T, Ohashi M,
Okudaira T, Yoshida Y, Wakimoto H, Hamada H,
Nakabayashi H, Tamaoki T, Omata M. In vivo genetherapy for alpha-fetoprotein-producing hepatocellular carcinoma by adenovirus-mediated transfer ofcytosine deaminase gene. Cancer Res 57 : 461-465,
1997.
13. Shiratori Y, Kato N, Yokosuka O, Imazeki F,
Hashimoto E, Hayashi N, Nakamura A, Asada M,
Kuroda H, Tanaka N, Arakawa Y, Omata M.
Predictors of the efficacy of interferon therapy inchronic hepatitis C virus infection. Gastroenterology
113 : 558-566, 1997.
14. Kojima H, Yokosuka O, Imazeki F, Saisho H, Omata
M. Telomerase activity and telomere length in hepatocellular carcinoma and chronic liver disease.
Gastroenterology 112 : 493-500, 1997.
15. Omata M. Treatment of chronic hepatitis B infection(Editorial). N Engl J Med 339 : 114-115, 1998.
16. Ono-Nita SK, Kato N, Shiratori Y, Lan KH, Yoshida
H, Carrilho FJ, Omata M. Susceptibility of lamivu-

167

dine-resistant hepatitis B virus to other reverse transcriptase inhibitors. J Clin Invest 103 : 1635-1640,
1999.
17. Yoshida H, Shiratori Y, Moriyama M, Arakawa Y, Ide
T, Sata M, Inoue O, Yano M, Tanaka M, Fujiyama S,
Nishiguchi S, Kuroki T, Imazeki F, Yokosuka O,
Kinoyama S, Yamada G, Omata M. Interferon therapy reduces the risk for hepatocellular carcinoma :National surveillance program of cirrhotic and noncirrhotic patients with chronic hepatitis C in Japan.
Ann Intern Med 131 : 174-181, 1999.
18. Shiratori Y, Imazeki F, Moriyama M, Yano M,
Arakawa Y, Yokosuka O, Kuroki T, Nishiguchi S,
Sata M, Yamada G, Fujiyama S, Yoshida H, Omata
M. Histologic improvement of fibrosis in patientswith hepatitis C who have sustained response tointerferon therapy. Ann Intern Med 132 : 517-524,
2000.
19. Kato N, Yoshida H, Ono-Nita SK, Kato J, Goto T,
Otsuka M, Lan KH, Matsushima K, Shiratori Y,
Omata M. Activation of intracellular signaling byhepatitis B and C viruses: C-viral core is the mostpotent signal inducer. Hepatology 32 : 405-412,
2000.
20. Otsuka M, Kato N, Lan KH, Yoshida H, Kato J, Goto
T, Shiratori Y, Omata M. Hepatitis C virus core protein enhances p53 function through augmentation of
DNA binding affinity and transcriptional ability. J
Biol Chem 275 :34122-34130, 2000.
21. Goto T, Kato N, Ono-Nita SK, Yoshida H, Otsuka M,
Shiratori Y, Omata M. Large isoform of hepatitis
delta antigen activates serum response factor-associated transcription. J Biol Chem 275 : 37311-37316,
2000.
22. Koike Y, Shiratori Y, Sato S, Obi S, Teratani T,
Imamura M, Yoshida H, Shiina S, Omata M. Des-rcarboxy prothrombin as a useful predisposing factor
for the development of portal venous invasion inpatients hepatocellular carcinoma -a prospective
analysis of 277 patients-. Cancer 91 : 561-569, 2001.
23. Ono-Nita SK, Kato N, Shiratori Y, Kato J, Goto T,
Schinazi RF, Carrilho FJ, Omata M. The polymerase
L528M mutation cooperates with nucleotide binding-site mutations, increasing hepatitis B virus replication and drug resistance. J Clin Invest 107 : 449455, 2001.
24. Yoshida H, Kato N, Shiratori Y, Otsuka M, Maeda S,
Kato J, Omata M. Hepatitis C virus corprotein activates nuclear factor κB-dependent signaling throughtumor necrosis factor receptor-associated factor. J
Biol Chem 276 : 16399-16405, 2001.
25. Yoshida H, Arakawa Y, Sata M, Nishiguchi S, Yano
M, Fujiyama S, Yamada G, Yokosuka O, Shiratori Y,
Omata M. Interferon therapy prolonged life
expectancy among chronic hepatitis C patients.
Gastroenterology 123 : 483-491, 2002.
26. Lan KH, Sheu ML, Hwang SJ, Yen SH, Chen SY, Wu
JC, Wang YJ, Kato N, Omata M, Chang FY, Lee SD.
HCV NS5A interacts with p53 and inhibits p53mediated apoptosis. Oncogene 21 : 4801-4811, 2002.

168

27. Shiratori Y, Shiina S, Teratani T, Imamura M, Obi S,
Sato S, Koike Y, Yoshida H, Omata M. Interferon
therapy after tumor ablation improves prognosis inpatients with hepatitis C viral hepatocellular carcinoma. Ann Inten Med 2002 (in press).

Gatroenterology
28. Tada M, Omata M, Ohto M. Clinical application of
ras-gene mutation for diagnosis of pancreatic adenocarcinoma. Gastroenterology 100 : 233-238, 1991.
29. Tada M, Omata M, Kawai S, Saisho H, Ohto M, Saiki
R, Sninsky J. Detection of ras gene mutations in pancreatic juice and peripheral blood of patients withpancreatic adenocarcinoma. Cancer Res 53 : 24722474, 1993
30. Ohta S, Yoshiura K, Takahashi M, Hata Y, Kohmoto
O, Kawabe T, Shimada T, Hiraishi H, Mutoh H,
Terano A, Sugimoto T, Omata M. P2 purinergic
receptor regulation of mucus glycoprotein by rabbit
gastric mucous cells in a primary culture.
Gastroenterology 106 : 1485-1492, 1994.
31. Shimada T, Ogura K, Ota S, Terano A, Takahashi M,
Hamada E, Omata M, Sumino S, Sassa R.
Identification of helicobacter pylori in gastric specimens, gastric juice, saliva and faeces of Japanesepatients. Lancet 343 : 1636-1637, 1994.
32. Takahashi M, Ota S, Shimada T, Hamada E, Kawabe
T, Okudaira T, Matsumura M, Kaneko N, Terano A,
Nakamura T, Omata M. Hepatocyte growth factor isthe most potent endogenous stimulant of rabbit gastric epithelial cell proliferation and migration in primary culture. J Clin Invest 95 : 1994-2003, 1995.
33. Tada M, Ohashi M, Shiratori Y, Okudaira T, Komatsu
Y, Kawabe T, Yoshida H, Machinami R, Kishi K,
Omata M. Analysis of K-ras gene mutation in hyperplastic duct cells of the pancreas without pancreaticdisease. Gastroenterology 110 : 227-231, 1996.
34. Imazeki F, Yokosuka O, Yamaguchi T, Ohto M, Isono
K, Omata M. Expression of variant CD44-messenger
RNA in colorectal adenocarcinomas and adenomatous polyps in humans. Gastroenterology 110 : 362368, 1996.
35. Lan KH, Kanai F, Shiratori Y, Okabe S, Yoshida Y,
Wakimoto H, Hamada H, Tanaka T, Ohashi M,
Omata M. Tumor-specific gene expression in carcinoembryonic antigen-producing gastric cancer cells
using adenovirus vectors. Gastroenterology 111 :
1241-1251, 1996.
36. Togo G, Toda N, Kanai F, Kato N, Shiratori Y, Kishi
K, Imazeki F, Makuuchi M, Omata M. A transforming growth factor beta type II receptor gene mutationcommon in sporadic cecum cancer with microsatellite instability. Cancer Res 56 : 5620-5623, 1996.
37. Takahashi M, Ota S, Hata Y, Mikami Y, Azuma N,
Nakamura T, Terano A, Omata M. Hepatocyte
growth factor as a key to modulate anti-ulcer actionof prostaglandins in stomach. J Clin Invest 98 : 26042611, 1996.
38. Lan KH, Kanai F, Shiratori Y, Ohashi M, Tanaka T,
Okudaira T, Yoshida Y, Hamada H, Omata M. In

vivo selective gene expression and therapy mediatedby adenoviral vectors for human carcinoembryonicantigen-producing gastric carcinoma. Cancer Res 57 :4279-4284, 1997.
39. Maeda S, Ogura K, Yoshida H, Kanai F, Ikenoue T,
Kato N, Shiratori Y, Omata M. Major virulence factors, VacA and CagA, are commonly positive in
Helicobacter pylori isolates in Japan. Gut 42 : 338343, 1998.
40. Maeda S, Yoshida H, Ikenoue T, Ogura K, Kanai F,
Kato N, Shiratori Y, Omata M. Structure of cag pathogenicity island in Japanese Helicobacter pylori isolates. Gut 44 : 336-341, 1999.
41. Asahi M, Azuma T, Ito S, Ito Y, Suto H, Nagai Y,
Tsubokawa M, Tohyama Y, Maeda S, Omata M,
Suzuki T, Sasakawa C. Helicobacter pylori Cag A protein can be tyrosine phosphorylated in gastric epithelial cells. J Exp Med 191 : 593-602, 2000.
42. Maeda S, Yoshida H, Ogura K, Mitsuno Y, Hirata Y,
Yamaji Y, Akanuma M, Shiratori Y, Omata M. H.
pylori activates NF-κB through a signaling pathway
involving IκB kinases, NF-κB-inducing kinase,
TRAF2, and TRAF6 in gastric cancer cells.
Gastroenterology 119 : 97-108, 2000.
43. Ogura K, Maeda S, Nakao M, Watanabe T, Tada M,
Kyutoku T, Yoshida H, Shiratori Y, Omata M.
Virulence factors of Helicobacter pylori responsible
for gastric diseases in Mongolian gerbil. J Exp Med
192 : 1601-1609, 2000.
44. Kawakami T, Chiba T, Suzuki T, Iwai K, Yamanaka
K, Minato N, Suzuki H, Shimbara N, Hidaka Y,
Osaka F, Omata M, Tanaka K. NEDD8 recruits E2ubiquitin to SCF E3 ligase. EMBO J 20 : 4003-4012,
2001.
45. Matsuhashi N, Watanabe K, Kanamori H, Ohnishi S,
Omata M. PPARgamma agonists for intestinal
ischaemia. Lancet 358 :75-76, 2001.
46. Mitsuno Y, Yoshida H, Maeda S, Ogura K, Hirata Y,
Kawabe T, Shiratori Y, Omata M. Helicobacter pylori
induced transactivation of SRE and AP-1 through the
ERK signalling pathway in gastric cancer cells. Gut 49:18-22, 2001.
47. Tateishi K, Omata M, Tanaka K, Chiba T. The
NEDD8 system is essential for cell cycle progression
and morphogenetic pathway in mice. J Cell Biol 155 :571-579, 2001.
48. Maeda S, Akanuma M, Mitsuno Y, Hirata Y, Ogura K,
Yoshida H, Shiratori Y, Omata M. Distinct mechanism of Helicobacter pylori-mediated NF-kappa B
activation between gastric cancer cells and monocytic cells. J Biol Chem 276 : 44856-44864, 2001.
49. Okamoto M, Shiratori Y, Yamaji Y, Kato J, Ikenoue T,
Togo G, Yoshida H, Kawabe T, Omata
M.Relationship between age and site of colorectal
cancer based on colonoscopy findings. Gastrointest
Endosc 55 : 548-551, 2002.
50. Hirata Y, Maeda S, Mitsuno Y, Akanuma M, Yanai A,
Yoshida H, Kawabe T, Shiratori Y, Omata M. H.
pylori CagA protein activates serum response ele-ment driven transcription independently of tyrosine
phosphorylation.Gastroenterology 2002 (in press).

169

Department of Nephrology
and Endocrinology
Outline and Research Objectives
The Department of Nephrology and Endocrinology is one of the major divisions in the
Department of Internal Medicine the University of Tokyo. It covers nephrology, endocrinology, and hypertension. In 1998, the Department of Internal Medicine of the University ofTokyo has been reorganized. Since then, Dr. Fujita has been Professor and Chairman of ourdepartment. His achievements touch on hypertension and sodium metabolism. This is oneof our main research objectives. The professor and each member of staff have an active
responsibility for all research, clinical, and educational activities. There are 12 AssistantProfessors supporting the clinical works of our residents in the ward, and also involved in
the Hemodialysis Unit. We are working side-by-side in all clinical activities under the supervision of the professor.
In our department, renal biopsy is actively performed and the morphrogical diagnosis of
the nephritis is established to give the real benefits of treatment to the patients. We also
treat diabetic patients with proteinuria and end-stage renal failure. Every staff physician ofour division also works at the hemodialysis unit thus we can manage patients in every stageof renal disease. The endocrine subunit attends a broad spectrum of endocrine entities providing the residents and academic internists a high quality of medical training. It is also our
speciality to diagnose and treat secondary hypertension caused by primary aldosteronism,
Cushing’s syndrome, pheochromocytoma and so on. We often have consultations from theother divisions concerning water and mineral metabolism disorders.
In our division, there are more than 30 students of the Graduate School. This suggests
high research activities and outstanding leadership of individual staff in our division.
Research seminars are held every Tuesday evening, where both senior investigators andtrainees present their own data and experimental plans and discuss profoundly with theprofessor and all members of our division. In the seminars, scientists outside our department and outside the University of Tokyo including foreign countries sometimes lecture
their papers, which encourages our research activities. Moreover, we are actively collaborating with some of the scientists outside our department. Our research topics are covered thealmost all field of hypertension, nephrology, and endocrinology.

Faculties and Students
Professor and Chair Toshiro Fujita. M.D., Ph.D.
Associate Professors Atsuo Goto, M.D., Ph.D.
Tomoki Okazaki, M.D., Ph.D.
Lecturers
Shigeo Taniguchi, M.D., Ph.D.
Kenmei Takaichi, M.D., Ph.D.
Assistant Professors ...............6 nephrologists and 6
endocrinologists
Post-doctorial fellows .............9
Graduate Students ................38
Research Students...................2
Secretaries ...............................2

Past Research and Major Accomplishments
The following major accomplishment has beenobtained using the best available molecular, physiological, and morphological techniques as possible.

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Vasoactive substances play an important role in
not only blood-pressure control but also formation ofvascular damages. Thus, we focused on
adrenomedullin (AM), a novel vasodilator peptide. To
elucidate physiological and pathophysiological function of AM, we generated AM knockout mice.
Homozygous AM knockout (AM-/-) mice were embryonic lethal but heterozygous (AM+/-) mice were grewnormally although its AM levels in plasma and tissues
were as a half as wild-type mice. Angiotensin II (AII)and salt loading caused severe coronary arterial damages in AM +/- mice. Moreover, AM +/- mice showedenhancement of cuff injury-induced intimal hyperplasia of femoral artery. These changes in AM+/- mice
were accompanied with enhanced oxidative stress,
suggesting that antioxidant effect of AM is importantin its protective effect against vascular damages. Thus,
intrinsic AM counteracts AII, and exhibits vasoprotective action.

Insulin resistance has been suggested to play animportant role in hypertension and the related organ
damages. Thus, we also focused on insulin resistancein several model animals of hypertension.
Interestingly, salt loading itself enhanced insulin
resistance in normotensive rats. Certainly, saltinduced rise in blood pressure in Dahl salt-sensitive
and angiotensin II (AII)-treated rats was accompaniedwith enhanced insulin resistance. The increased
insulin resistance in hypertensive models is intimately related to oxdative stress. Interestingly, aged AM
deficient mice showed insulin resistance, associatedwith overproduction of oxidative stress. Then, this is
the model of insulin resistance syndrome, so-called“syndrome X”.
Oxidized low density lipoprotein (OxLDL) has
been implicated in pathophysiology of atherosclerosis.
OxLDL alters endothelial function and triggered
atherogenic changes. Thus, we focused on a novel
endothelial OxLDL receptor, lectin-like OxLDL receptor-1 (LOX-1). We observed that LOX-1 was enhanced
in aortas and kidneys of rats with hypertension, diabetes, and renal failure. Also, LOX-1 was stimulatedby oxdative stress, which is known to cause organ
damages by common diseases. The expression of
LOX-1 was intimately related to nuclear factor (NF)kB. Thus, LOX-1 was implicated in pathphysiology ofatherosclerosis and glomeruloscrelosis via oxidative
stress.
To elucidate a novel candidate gene for salt sensitiviy of blood pressure, we performed differential display assay between Dahl salt-sensitive (DS) and–resistant (DR) rats. As a result, we found REPT1 (ratretrovirus-like element transcribed in proximal tubulus) in the kidneys of DS rats. REPT1 was locatd in
the proximal tubules and its renal expression was correlated with salt sensitivity among different strains ofrats.
Intracellular calcium ion (Ca2+) is a spatially andtemporally organized second messenger, which mediates a variety of important functions of vascularendothelial cells ranging from the production ofvasoactive substances to gene expression. We developed FRET-based technique to image localized Ca2+
signaling in live endothelial cells. And, we succeeded
to elucidate localized Ca2+ signaling and function of
caveolae. Endothelial nitric oxide (NO) was localizedin caveolae and activity of NO was regulated by intracellular Ca2+ and binding to caveolae.
G proteins control a diverse network of signals.
We have focused on mutation of G proteins in humandiseases. We have discovered how point mutations inaS (the a subunits of GS) alter GS signaling, causing
human diseases. Also, we succeeded in building up a
simple cell-free method to study interaction between
receptors and G proteins. We have, for the first time,achieved success in designing dominant negative
mutant Gα proteins in realistic level. Also, we analyzed relationship of C825T polymorphism of Gβ3 tohypertension, which has been demonstrated in
Caucasian people.
We revealed that ionic mechanisms such as activation of inwardly-rectifying potassium currents andactivation of nonselective cation currents are important in the regulation of hormone secretion from pituitary cells using electrophysiological techniques. Also,
we elucidated the mechanisms of action of regulatorsof circulation system such as AM, proadrenomedullin
N-terminal 20 peptide (PAMP), and AII. In addition,
we studied mineralocorticoid action in pathophysiology of cardiovascular diseases.
We studied molecular basis for diseases of calcium
2+(Ca ) metabolism. For example, we have identified
novel mutations in Ca-sensing receptor gene inpatients with Type ⅡBartter ‘s syndrome as well as
those with familial hypocalciuric hypercalcemia(FHH). Also, we characterized proteoglycans synthesized by osteoblasts biochemically and explored indetail cell-matrix interactions in osteoblasts. In addition, we identified negative Ca2+ responsive element(nCaRE) in the parathyroid hormone (PTH) gene.
From a physiological standpoint, we elucidated thebasic transport properties of sodium-bicarbonate (Na+HCO3-) cotransport (NBC1) in renal proximal tubularacidosis (RTA) with ocular abnormalities. This is an
example of a successful translation of basic finding
into clinical medicine. We clarified the regulatory
mechanism of proximal transport in physiological andpathological states and demonstrated the cAMP-activated chloride (Cl-) conductance in proximal tubules.
We revealed the immunohistochemical localizationand function of Kir 7.1 potassium channel, organic
anion transporter (OAT1), and stanniocalcin in the rat
kidney. We cloned the Na + /H + antiporter (NHE)
cDNAs from human tissues and characterized the regulation of the expression of NHE in kidney cells.
With the application to acute renal failure or graft
survival in mind, we demonstrated the successfulintervention by cyclic RGD (Arg-Gly-Asp) peptide to
reduce the renal ischemia-reperfusion model and the
diagnistic utilities of monitoring the radio-labbeled
RGD peptides. We demonstrated the incorporation ofoligonucleotides to proximal tubular epithelial cells
and the amelioration of renal ischemia-reperfusion
injury using antisense oligonucleotides specificiallytargeted to inducible NO synthase (NOS). We showedby in vivo approach the chemotactic activity ofleukotriene B4 to drive neutrophil in acute ischemiareperfusion injury.
The clarification of the mechanism of the progressive renal disease is the major problem to be solved in
the field of nephrology. We contribute much to the171solution of immunological mechanism involved inglomerulonephritis, especially in terms of complement and complement regulatory proteins. We
revealed the localization of NOS and NAD(P)H oxidase isoforms in the kidney and gave insight into thefunctional role of NO and oxidative stress in the kidney of hypertension, glomerulonephritis, and diabetes. We examined the role of physical factors suchas mechanical stretch on TGF-β and extracellularmatrix accumulation in the progression of renal disease. We demonstrated that renoprotective C-typenatriuretic peptide was produced in renal parenchymal cells.
We isolated several transcription factors from vascular myocytes such as an isoform of MEF2A and
GATA-6 and identified the implication of these factors
in the differentiation or dedifferentiation of vascular
myocytes and vascular hypertrophy process. We clarified the intracellular pathway involved in the cell
cycle progression with endothelin-1 proved to promote the renal injury.
As a new technique, we developed a simple andreliable method of quantitating mRNAs in minute tissue samples in addition to the established microdissection and single transduction mechanism by whichvasopressin and prostaglandins interact in the kidneyusing immunodissected tubule. We also developed anovel animal model of glomerular endothelial injury,
which may help elucidate the mechanism of glomerular endothelial damage.
Research on renal regeneration has just started to
do the identification of adult stem cells in the kidney.
Using a flow cytometry assay and Hoechst 33342
staining, we isolated adult stem cells in the kidney asside population (SP) cells that have been identified asstem cells in bone marrow, skeletal muscle and liver.
Using 3 dimensional culture systems, moreover, cultured renal SP cells differentiate into various kinds ofcells.
Troughout our research we have attempted to link
our basic finding with our clinical work-----between
bench and bedside with either direction.

Current Research
To extent our past accomplishment further, we are
now attempting to clarify the following themes.
Recent studies have revealed that oxidative stressis intimately related to not only hypertension but also
the other common disease such as diabetes andhypercholesterolemia, so we are examining whether
these abnormalities including insulin resistance develop AM+/- mice, in which oxidative stress is enhanced.
Also, we are investigating the effect of hypoxia in
AM +/- mice, because hypoxia-induced damage isknown to relate to oxidative stress.

172

To elucidate atherogenic role of LOX-1, we are
studying whether atherogenic stimuli such as AII
infusion and cuff injury cause severe vascular damages in LOX-1 transgenic mice. Moreover, we foundthat peroxisome proliferator-activated receptor γ(PPARγ) ligands inhibit cytokine-stimulated LOX-1
expression in vitro and in vivo. Now we are studying
this mechanism, which may suggest that the inhibition of LOX-1 might be related to antiatherogenic
effect of PPARγ ligands.
We developed FRET-based tool to image subplasmalemmal Ca2+ and cAMP. We are now studying roleof caveolin scaffolding domain and hsp90 on NO production using unique cell biological and imaging techniques.
By the cell-free method to study interaction
between receptors and G proteins, we studied thespecificity between many G proteins and G proteincoupled receptors for cardiovascular hormones, suchas AII, catecholamines, and AM.
We are studying the central role of Ca-sensing
receptor in salt-sensitive hypertensive model animal,
in which Ca loading ameliorated hypertension.
Moreover, we have many themes such as electrophysiology of pituitary, signal transduction of mineralocorticoid receptor, mechanisms of age-related osteopenia,
and so on.
We further clarify molecular mechanism of proximal RTA based on newly identified NBC1 mutation.
The molecular basis of ion transport in NBC1 is studying via giant patch technique.
We are studying roles of lipid mediators in renal
ischemia-reperfusion injury using several lines of micedeleted/overexpressed lipid mediator genes. We are
analyzing detailed intracellular pathways implicatedin the expression of monocyte chemoattractant protein-1 in vascular myocytes.
We are investigating role of endogenous vasoactive
substances in the progression of renal diseases suchas AII, AM, leukotriene, retinoid and glucocorticoids
using knockout mice and renal cells. We are also
studying roles of oxidative and nitrosoactive stress in
the renal diseases including diabetic nephropathy,
ischemia-reperfusion injury, and long-term hemodialysis complications.
Now 3 projects of renal regeneration are on going.
First, optimizing the 3 dimensional culture systems todifferentiate renal SP cells to metanephros. Second,clarifying the condition and identifying the morphogen to differentiate human adult stem cells suchas bone marrow derived mesenchymal stem cells into
renal components. Third, investigating the existenceof stem cell dysfunction in patients with renal dysfunction, and getting comprehensive information forintervention using DNA microarray analysis.

Future Prospects
The main goal of our studies is to cure patientswith renal and endocrinological diseases and hypertension to reduce the number of patients with hypertensive complications and renal failure. To serve thispurpose, we have to focus our effort to clarify themechanisms of diseases. There are not only numerous
diseases in our field but also multiple mechanisms to
involved especially in chronic stage of renal failureand caidiovascular complication-associated hypertension. Thus, we are applying the multi-dimentional
approach such as oxidative stress, lipid metabolism,
hemodynamics, and vasoactive substabces. These
approaches will lead to the development of several
new drugs and strategies to treat renal and endocrinological diseases, hypertension, and its complications.
By the discoveries of the candidate genes for salt-sensitive hypertension, we can try to choose the suitableantihypertensive therapy and salt restriction for individual patients (tailor-made medicine).

Research Grants
1. Toshiro Fujita: 1998-2002 Grant-in-aid for scientificresearch from the Ministry of Education, Culture,
Sports, Science and Technology of Japan (specificfield): Molecular biological and embryological investigation of adrenomedullin, 54,5000,000 yen
2. Toshiro Fujita: 2000-2002 Grant-in-aid for scientificresearch from the Ministry of Education, Culture,
Sports, Science and Technology of Japan (B) Role ofoxidized LDL/lectin-like oxidized LDL receptor-1(LOX-1) system in the development of glomerulosclerosis. 15,000,000 yen
3. Atsuo Goto: 1995-1997 Grant-in-aid for scientificresearch from the Ministry of Education, Culture,
Sports, Science and Technology of Japan (B) Role ofendogenous digitalis-like substance in relationshipbetween stress and cardiovascular diseases,
7,500,000 yen
4. Tomoki Okazaki: 1999-2001 Grant-in-aid for scientificresearch from the Ministry of Education, Culture,
Sports, Science and Technology of Japan (B): The
mechanism of transcriptional inhibition by nuclear
hormone receptors by using a viamine D receptor asa model, 14,000,000 yen
5. Taro Iiri (Assistant professor of endocrinology): 20022004 Grant-in-aid for scientific research from the
Ministry of Education, Culture, Sports, Science andTechnology of Japan (A): G protein and diseases:
Analysis of molecular mechanism and regulation of
signal transduction, 30,000,000 yen

Select Publications
1. Watanabe S, Fukumoto S,Chang H, Takeuchi Y,
Hasegawa Y, Okazaki R, Chikatsu N,Fujita T:

Association between activating mutations of calcium-sensing receptor and Bartter’s syndrome.
Lancet,360:692-694,2002
2. Ogihara T, Asano T, Ando K, Sakoda H, Anai M.
Shojima N, Ono H, Onishi Y, Fujimoto M, Abe M.
Fukushima Y, Kikuchi M, Fujita T: High-Salt Diet
Enhances Insulin Signaling and Induces Insulin
Resistanse in Dahl Salt-Sensitive Rats, Hypertension,
2002;40:83-89.
3. Shimosawa T. Shibagaki Y. Ishibashi K. Kitamura K.
Kangawa K. Kato S. Ando K. Fujita
T:.Adrenomedullin, an endogenous peptide, counteracts cardiovascular damage. Circulation. 105(1):10611, 2002.
4. Sun D. Seki G. Uwatoko S. Nakao A. Goto A. Fujita T.
Kimura S. Taniguchi S: Quantifying porphobilinogendeaminase mRNA in microdissected nephron segments by a modified RT-PCR. Kidney International.
61(1):336-41, 2002.
5. Onozato ML. Tojo A. Goto A. Fujita T:. Wilcox CS:
Oxidative stress and nitric oxide synthase in rat diabetic nephropathy: effects of ACEI and ARB.Kidney
International. 61(1):186-94, 2002.
6. Chabrashvili T, Tojo A, Onozato M, Kitiyakara C,
Quinn M, Fujita T: Welch W, Wilcox S : Expression
and cellular localization of classic NADPH oxidasesubunits in the spontaneously hypertensive rat kidney. Hypertension 39:269-274,2002.
7. Suzuki E, Nishimatsu H, Satonaka H, Walsh K, Goto
A, Omata M, Fujita T: Angiotensin Ⅱ induces
myocyte enhancer factor 2- and calcineurin/nuclear
factor of activated T call-dependent transcriptional
activation in vascular myocytes. Circ Res. 90:10041011,2002.
8. Usui T, Hara M, Satho H, Moriyama N, Kagaya H,
Amano S, Oshita T, Ibaraki N, Hara C, Kunimi M,
Norii E, Tsukamoto K, Inatomi J, Kawakami H,
Endou H, Igarashi T, Goto A, Fijita T, Araie M:
Molecular basis of ocular abnormalities associatedwith proximal renal tubular acidosis. The Journal of
Clinical Investigation.108:107-115,2001.
9. Chiba Y, Ogita T, Ando K, Fujita T: PPARγ ligands
inhibit TNF-α-induced LOX-1 expression in cultured endothelial cells. Biochem Biophys Res
Commun 286: 541-546, 2001.
10. Nagase M, Ando K, Nagase T, Kaname S, Sawamura
T, Fujita T: Redox-sensitive regulation of LOX-1 gene
expression in vascular endothelium. Biochem
Biophys Res Commun 281: 720-725, 2001.
11. Chang H. Fujita T: PicSNP:a browsable catalog ofnonsynonymous single nucleotide polymorphisms in
the human genome. Biochem Biophys Res Commun287(1):288-91, 2001.
12. Sogabe H. Nangaku M. Ishibashi Y. Wada T. Fujita T.
Sun X. Miwa T. Madaio MP. Song WC: Increased
susceptibility of decay-accelerating factor deficient
mice to anti-glomerular basement membrane
glomerulonephritis. Journal of Immunology.
167(5):2791-7, 2001.

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13. Sekine N. Fukumoto S. Ishikawa T. Okazaki T. Fujita
T: GH inhibits interferon-gamma-induced signaltransducer and activator of transcription-1 activationand expression of the inducible isoform of nitricoxide synthase in INS-1 cells. Endocrinology.
142(9):3909-16, 2001.
14. Shao J. Miyata T. Yamada K. Hanafusa N. Wada T.
Gordon KL. Inagi R. Kurokawa K. Fujita T. Johnson
RJ. Nangaku M: Protective role of nitric oxide in amodel of thrombotic microangiopathy in rats.
Journal of the American Society of Nephrology.
12(10):2088-97, 2001.
15. Noiri E. Nakao A. Uchida K. Tsukahara H. Ohno M.
Fujita T. Brodsky S. Goligorsky MS: Oxidative andnitrosative stress in acute renal ischemia: American
Journal of Physiology - Renal Fluid & Electrolyte
Physiology. 281(5):F948-57, 2001.
16. Watanabe S. Fukumoto S. Takeuchi Y. Fujita H.
Nakano T. Fujita T: Effects of 1-year treatment withfluvastatin or pravastatin on bone. American Journalof Medicine. 110(7):584-7, 2001
17. Nagase M. Ando K. Nagase T. Kaname S. Sawamura
T. Fujita T:.Redox-sensitive regulation of lox-1 gene
expression in vascular endothelium. Biochem
Biophys Res Commun 281(3):720-5, 2001.
18. Kanda H. Kubo K. Hamasaki K. Kanda Y. Nakao A.
Kitamura T. Fujita T. Yamamoto K. Mimura T:
Influence of various hemodialysis membranes on theplasma (1-->3)-beta-D-glucan level. Kidney
International. 60(1):319-23, 2001
19. Yamada K. Hori Y. Hanafusa N. Okuda T. Nagano N.
Choi-Miura NH. Couser WG. Miyata T. Kurokawa K.
Fujita T: Nangaku M. Clusterin is up-regulated inglomerular mesangial cells in complement-mediatedinjury. Kidney International. 59(1):137-46, 2001.
20. Suzuki J. Ohnsihi H. Shibata H. Wada A. Hirayama
T. Iiri T. Ueda N. Kanamaru C. Tsuchida T. Mashima
H. Yasuda H. Fujita T: Dynamin is involved inhuman epithelial cell vacuolation caused by the
Helicobacter pylori-produced cytotoxin VacA.
Journal of Clinical Investigation. 107(3):363-70, 2001.
21. Kanamaru C. Yasuda H. Takeda M. Ueda N. Suzuki J.
Tsuchida T. Mashima H. Ohnishi H. Fujita T: Smad7
is induced by norepinephrine and protects rat hepatocytes from activin A-induced growth inhibition.
Journal of Biological Chemistry. 276(49):45636-41,
2001
22. Chang H. Fujita T: A numerical model of acid-basetransport in rat distal tubule. American Journal of
Physiology - Renal Fluid & Electrolyte Physiology.
281(2):F222-43, 2001.
23. Ishikawa T, Sekine N, Hata K, Igarashi T, Fujita T:
Prostagladin A1 enhances c-fos expression and activating protein-1 activity. Molecular and Cellular
Endocrinology 164:77-85,2000.
24. Sekine N, Ishikawa T, Okazaki T, Hayashi M,
Wollheim C.B., Fujita T: Synergitic activation of NFκB and inducible isoform of nitric oxide synthaseinduction by interferon-γ and tumor necrosis factor-α in INS-1 cells. J.Cell Physiol.184:46-57,2000.

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25. Chikatsu N, Takeuchi Y, Tamura Y, Fukumoto S,
Yano K, Tsuda E, Ogata E, Fujita T: Interactions
between cancer and bone marrow cells induce osteoclast differentiation factor expression and osteoclastlike cell formation in vitro. Biochem Biophys Res
Commun 267:632-637,2000.
26. Noiri E, Yokomizo T, Nakao A, Izumi T, Fujita T,
Kimura S, Shimizu T: An in vivo approach showing
the chemotantic activity of leukotrene B4 in acuterenal ischemic-reperfusion injury. ProcNatl Acad Sci.
97:823-828,2000.
27. Chikatsu N, Fukumoto S, Takeuchi Y, Suzawa M,
Obara T, Matsumoto T, Fujita T: Cloning and characterization of two promoters for the human calciumsensing receptor(CaSR) and changes of CaSR expression in parathyroid adenomas. J Biol Chem 275:75537557,2000.
28. Kunimi M, Seki G, Hara C, Taniguchi S, Uwatoko S,
Goto A, Kimura S, Fujita T: Dopamine inhibits renal
Na＋:HCO3- contransporter in rabbits and normotensive rats but not in spontabeously hypertensive rats.
Kidney International 57:534-543,2000.
29. Kubota N, Terauchi Y, Miki H, Tamemoto H,
Yamauchi T, Komeda K, Satoh S, Nakano R, Ishii C,
Sugiyama T, Eto K, Tsubamoto Y, Okuno A
Murakami K, Sekihara H, Hasegawa G, Naito M,
Toyoshima Y, Tanaka S, Shiota K, Kitamura T, Fujita
T, Ezaki O, Aizawa S, Nagai R, Tobe K, Kimura S,
and Kadowaki T.: PPARγ mediates high-fat dietinduced adipocyte hypertrophy and insulin resistance. Mol Cell. 4:597-609,1999.
30. Hori Y, Yamada K, Hanafusa N, Okuda T, Okada N,
Miyata T, Couser W, Kurokawa K, Fujita T, Nangaku
M: Crry, a complement regulatory protein, modulates
renal interstitial disease induced by proteinuria.
Kidney International 56:2096-2106,1999.
31. Chang H, Fujita T: A kinetic model of the thiazidesensitive Na-Cl cotransporter. Am J Physiol
276:F952-F959,1999.
32. Chang H, Fujita T: A numerical model of the renal
distal tubule.Am J Physiol 276:F931-F951,1999.
33. Hayashi M, Shimosawa T, Fujita T: Hyperglycemia
increases vascular adrenomedullin expression.
Biochem Biophys Res Commun 258: 453-456,1999.
34. Yasufuku-Takano J, Takano K, Takei T, Fukumoto S,
Teramoto A, Takakura K, Yamashita N, Fujita T :Heterozygous gsp mutation renders ion channnels ofhuman somatotroph adenoma cells unresponsive to
growth hormone-releasing hormone. Endocrinology
140: 2018-2026, 1999.
35. Suzuki Y, Itakura M, Kashiwagi M, Nakamura N,
Matsuki T, Sakuta H, Naiyo N, Takano K, Fujita T,
Hirose S: Identification by differential display of a
hypertonicity-inducible inward rectifier potassiumchannnel highly expressed in chloride cells. J
Biological Chem 274: 11376-11382,1999.
36. Suzawa M, Takeuchi Y, Fukumoto S, Kato S, Ueno N,
Miyazono K, Matsumoto T, Fujita T: Extracellularmatrix-associated bone morphogenetic proteins areessential for differentiation of murine osteoblastic
cells in vitro. Endocrinology 140: 2125-2133, 1999.
37. Takano K, Fujita T : Proadrenomedullin N-terminal
20 peptide hyperpolarizes the membrane by activating an inwardly rectifying K+ cyrrent in differentiated
PC12 cells.Circ Res. 84:445-450,1999.
38. Nagase M, Kato A, Ono T, Suzuki Y, Hirose S, Fujita
T : Retrotransposons transcribed preferentially inproximal tubules of salt-hypertensive rats. Kidney
International 55: 995-1004, 1999.
39. Okazaki R, Chikatsu N, Nakatsu M, Kakeuchi Y,
Ajima M, Miki J, Fujita T, Arai M, Totsuka Y,
Tanaka K, Fukumoto S :A novel activating mutationin calcium-sensing receptor gene associated with afamily of autosomal dominant hypocalcemia. J Clin
Endocrinol Metab 84: 363-366,1999.
40. Iiri T, Bell S.M., Baranski T.J., Fujita T, Bourne H.R.
:A Gsα mutant designed to inhibit receptor signaling
through Gs. Proc Natl Acad Sci. 96: 499-504, 1999.
41. Iiri T, Bell S.M., Baranski T.J., Fujita T, Bourne H.R.:A Gsα mutant designed to inhibit receptor signaling
through Gs. Proc Natl Acad Sci. 96: 499-504, 1999.
42. Isshiki M, Ando J, Korenaga R, Kogo H, Fujimoto T,
Fujita T, Kamiya A: Endothelial Ca2+ waves preferentially originate at specific loci in caveolin-rich celledges. Proc Natl Acad Sci 95:5009-5014,1998.
43. Nagase M, Abe J, Takahashi K, Ando J, Hirose S,
Fujita T : Genomic organization and regulation of
expression of the lectin-like oxidized low-density
lipoprotein receptor(LOX-1) gene. J Biological Chem
273:33702-33707,1998.
44. Hayashi M, Shimosawa T, Isaka M, Yamada S, Fujita
R, Fujita T: Plasma adrenomedullin in diabetes.
Lancet 350: 1449-1450, 1997.
45. Nakaoka T, Gonda K, Ogita T, Otawara-Hamamoto
Y, Okabe F, Kira Y, Harii K, Miyazono K, Takuwa Y,
Fujita T: Inhibition of Rat Vascular Smooth Muscle
Proliferation in Vitro and in Vivo by BoneMorphogenetic Protein-2. J Clin Invest 100: 28242832,1997.
46. Takano K, Yasufuku-Takano J, Teramoto A, Fujita T:
Corticotropin-releasing hormone excites adrenocorticotropin-secreting human pituitary adenoma cells by
activating a nonselective cation current. J Clin Invest
98:2033-2041,1996.
47. Takano K, Yamashita N, Fujita T: Proadrenomedullin
NH2-terminal 20 peptide inhibits the voltage-gated
Ca2+ channel current through a pertussis toxin-sensitive G protein in rat pheochromocytoma-derived PC
12 cells. J Clin Invest 98:14-17,1996.
48. Chung U, Tanaka Y, Fujita T: Association ofInterleukin 6 and hypoaldosteronism in patientswith cancer. New Engl J Med 334:473,1996.
49. Shimosawa T, Ito Y, Ando K, Kitamura K, Kangawa
K, Fujita T: Proadrenomedullin N-terminal 20 peptide, a new product of the adrenomedullin gene,
inhibits norepinephrine overflow from nerve endings. J Clin Invest 96:1672-1676,1995.
50. Kitagawa M, Chang H, Fujita T: Hyperkalemia due tonafamostat mesilate. New Engl J Med 332:687,1995

175

Department ofAllergy and Rheumatology
Outline and Research Objectives
Our department was established in 1998 according to the reorganization of the Internal
Medicine. Doctors from previous Department of Medicine and Physical therapy and theThird Department of Internal Medicine have joined our department. Our main clinical field
are rheumatology and allergy. Therefore, our research fields also cover the areas including
basic immunology, clinical immunology, rheumatology and allergy. Especially, an area called
applied immunology such as developing new therapeutic methods has ready became important and some projects in these directions have thus been carried out in our department.
Our Reumatology and Immunology field comprises several projects including T cell receptor, autoantibody, oral tolerance, cell signaling and genetic analysis. The allergy field comprises projects of airway inflammation and IgE response.

Faculties and Students
Professor and Chair

Kazuhiko Yamamoto, M.D.
(Rheumatology, Allergology)1997
Lecturer Hirokazu Okudaira, M.D. (Allergology)
Fujio Takeuchi, M.D. (Rheumatology)
Yosikata Misaki, M.D. (Rheumatology)
Zen-ichiro Honda, M.D.(Rheumatology)
Associate .................................8
Postdoctoral Fellow ................7
Graduate student ..................16
Research student.....................2
Secretary .................................4

Past Research and Major Accomplishments
Rheumatology and Immunology
1. T Cell Receptor (TCR) Project
We have established a method for analyzing T-cellclonality using RT-PCR of TCR messages and subsequent electrophoretic separation of the PCR productbased on their single strand conformation polymorphism (SSCP). Using this method an analysis of PBLs
obtained from a healthy individual revealed the heterogeneity of the CDR3 region of the TCR beta messages. On the other hand, clonally accumulated T cells
could be identified as distinct bands on the background heterogeneous electrophoretic patterns. With
this method, we reported the importance of antigenspecific T cell clones in the pathogeneses of RA, SLE,
other autoimmune diseases.
Having observed the importance of antigen-specific T cells in autoimmune disorders, we are now trying

176

to establish methods to generate regulatory T cells
engineered by transfer of genes, the information ofwhich could be obtained from an individual patient
sample. At first, we generated an efficient alpha/beta
TCR gene transfer system using two independent
monocistronic retrovirus vectors. This system enabled
us to express the clonotypic TCR in about 25-40% of
the CD4+ T cells from murine splenocytes. The transduced cells showed an antigen-specific proliferation invitro as well as an antigen-specific delayed-type
hypersensitivity in vivo. Furthermore, regulatory T
cells engineered by the transfer of collagen-specific
TCR and IL-10 genes suppressed collagen induced
arthritis in mice. In addition, nucloesome-specific T
cells generated by the similar triple gene transfer of
TCR and CTLA4Ig were applied to lupus-prone
NZB/W F1 mice. These cells suppressed the related
pathogenic autoantibody production and nephritis
without impairing the T cell-dependent humoral
immune responses. Thus, from these results, we
could obtain efficient TCR function by gene transfer.
2. Autoantibody Project
We have been investigating in the regulatory
mechanism of autoantibody production, which is thehallmark of systemic autoimmune diseases such assystemic lupus erythematosus. We generated a transgenic mice which expresses human U1snRNP-A
autoantigen. We found that splenic T cells from thistransgenic mice suppressed the immune response tohuman U1-A. In order to further examine the regulation of autoreactive T cell in a clone level, We also
generated a transgenic mice expressing ovalbumin(OVA) systemically in nucleus. In the combination
with OVA-specific T cell receptor transgenic mice, wedemonstrated that dendritic cells are able to tolerize
autoreactive T cells. Furthermore, we found that selfreactive T cells matured into CD4(+)CD25(+) regulatory T cells in the thymus.

2) Linkage analysis using a familial juvenile hyperuicemic nephropathy (FJHN) revealed the susceptibility region in chr16p12.

3. Oral Tolerance Project
We have studied the mechanism of tolerance
induction in the mucosal immune system and its
application to therapy of autoimmune diseases.
Especially, we focused on the role of DC (dendritic
cells) for oral tolerance after antigen feeding. We
examined the requirement for tolerance induction in
adoptive cell transfer system and found that not only
T cells but DCs in Peyer’s patches or spleen are important for the transfer of oral tolerance. We also fed
OVA to Balb/c mice in bronchial asthma mouse model
induced by OVA and confirmed that OVA feeding
suppressed airway hypereactivity. In this system,
depletion of DCs abrogated the transfer of tolerance.

Allergy
1. Airway Inflammation Project
We established a system to evaluate antigen specific response of the airway in the mouse under a noninvasive, unrestrained condition. Using this system,
we analyzed relationships among sensitization to anallergen, increase in airway hyperreactivity (AHR),
and development of airway inflammation. The results
demonstrated that contrary to the previous general
understandings, AHR can be induced with systemic
sensitization alone (early phase AHR).
We have also found that intratracheal administration of hepatocyte growth factor (HGF), a regenerative
growth factor for bronchial and alveolar epithelial
cells could attenuate extent of the experimental lungfibrosis. In addition, we elucidated the mechanism ofaction and found that HGF has a potent migratory
activity and enhancing activity of surface fibrinolytic
system of lung epithelial cells. Further, we reported
that anti-fibrotic cytokine IFN-γ potentiates the biological activity of HGF by upregulating the expressionof c-Met/HGF receptor on alveolar epithelial cells.

4. Intracellular Signaling Project
Receptors Fc portion of Immunoglobulines play
central roles in the initial step of immune regulation.
For the cells to be activated, the receptors should becross-linked by multivalent ligands. We have engaged
in solving the mechanisms, in which physical receptor
cross-linking is converted into biochemical signaling.
It was assumed that receptor engagement is sensed
and converted by Src family kinases (SFKs) to cellinner events. However, the compensatory function ofeight SFKs hampered clear demonstration of thenotion. We introduced novel strategies to circumventthe difficulty, and established their requirement andspecificity in Fc, and c-Kit and Integrin receptors.
Notably, SFKs responsible for Fc receptor signaling are
all modified with palmitic acid, and they reside in amembrane domain referred to as ‘lipid rafts’.
On the other hand, we found that a 105 kDa
moledule was tyrosine phosphorylated through TCR
ligation in human T cells. It was proven to be p105CasL,
a member of the p130Cas-related docking protein family, which subsequently binded to the Src homology 2
domain of c-Crk. We showed that it was also tyrosine
phosphrylated though integrin stimulation in T cells,
and its expression was predominant in lymphocytes.
Next, we reported that Fyn and Lck tyrosine kinases
regulated tyrosine phosphorylation of p105CasL in TCRmediated signaling in human T cells and splenocytesof lpr mice, SLE model mice.
5. Genetics Project
We have studied genetic background and pathophysiology of rheumatic diseases, and mechanisms
and effect of aging were studied. Examples are1)Genetic analysis of HLA-DRB1 shared epitope in
RA family cases showed the participation of the epitope was neither unique nor indispensable.

2. IgE Response Project
Surface expression of high affinity IgE receptor(FceRI) on mast cells is essential for these cells toexert IgE-dependent effector functions. We have analyzed how and to what extent mast cell surface FceRI
expression is regulated, and whether regulation of
their surface FceRI levels is functionally important.
Through the series of our study, we found that the ligand of FceRI, IgE, which is known to stabilize FceRI,
can greatly upregulate the surface FceRI levels onmast cells. A Th2 cytokine, IL-4, is also involved in
FceRI upregulation in mast cells. Mast cells withupregulated levels of surface FceRI in the presence of
IgE and/or IL-4 demonstrate enhancement in IgEdependent mediator release and cytokine production.
On the other hand, glucocorticoid, decreases the surface FceRI levels of mouse mast cells either in thepresence or absence of IgE, via posttranscriptional
mechanism. These results indicate that mast cell surface FceRI levels are regulated via multiple mechanisms by various factors including IgE, cytokines andglucocorticoids, resulting in functional alteration ofmast cells. IgE- and FceRI-related positive feedback
mechanism may account for the pathogenesis of IgEmediated allergy.

177Current Research
Rheumatology and Immunology
1. TCR Project
We are now trying to establish an efficient method
to reconstitute TCR function using information available from a small sample of a patient. In order toaccomplish these projects, two important technological breakthroughs are necessary. They are cloning
techniques of full length cDNA encoding alpha andbeta TCR from a single cells and efficient vectors todeliver these genes into usual lymphocytes. We are
now trying to develop several possible technicalimprovements.
2. Autoantibody Project
We have identified several unique molecules to
CD4(+)CD25(+) regulatory T cells including transcription factors and proliferation-associated molecules,
which indicate that the subset is a unique and distinct
population.
3. Oral Tolerance Project
We are studying the mechanisms of generation ofregulatory T cells which mediate mucosal tolerance.
We are visualizing the interaction between T cells and
DCs which captured antigen in Peyer’s patch and characterizing DCs which are involved in generation ofregulatory T cells. We are also investigating a role of
TGF-β for generation of regulatory cells in mucosal
tolerance and for class switching of B cells to IgA
secreting plasma cells.
4. Intracellular Signaling Project
We are searching for the molecules responsible for
Fc receptor-mediated lipid raft assembly through protein chemistry approaches. We have also cloned two
novel molecules interacting with a SFK, Lyn. One isan ubiquitin ligase with RING domain, presumably
promoting SFK turnover. Another is a huge adaptor
protein possessing putative Grb2 and PI3K bindingsites and GAP domain.
We are also investigating whether IGF-I could prevent glomerulosclerosis, because glomerulosclerosis
was not found in IGF-I overexpressing mice, althoughglomerulohypertrophy became obvious. To investigate
this, we are evaluating the effects of adding IGF-I after
adhesion stimulation on some signaling molecules in
rat mesangial cells. Concurrently, we are preparing an
animal study, using 5/6 nephrectomy as glomeruloscrelosis model rat, to know whether IGF-I infusion
prevents glomerulosclerosis.
5. Genetics ProjectSome candidate gene analysis are now in progress

178

Allergy
1. Airway Inflammation Project
We have recently found that passive cell transferof spleen cells obtained from sensitized mouse into
naive mouse provokes an increase in the AHR. This
indicates that mechanisms to induce AHR can bestudied. We are now analyzing which kind of cell in
the lung, such as antigen presenting cells, mast cells,
and lymphocytes, would contribute to the development of early phase AHR by negative and positive
selection of spleen cells.
2. IgE Response Project
We are interested whether IgE crosslinking stimulation of mast cells and basophils can result in cellularchanges other than activation. Currently we are
exploring the phenomenon of FceRI-mediated desensitization. At present we think that cellular desensitization and stimulation may be separate but closely
related to each other.

Future Prospects
Rheumatology and Immunology
1. TCR Project
Our final goal is to establish an antigen- specificimmunotherapy in terms of TCR gene transfer using
information from a small sample of an individual
patient. Once the basic system is established, we
could apply this to several different immunologicaldisorders. Applicable disorders include rheumatoid
arthritis, systemic lupus erythematosus and otherconnective tissue diseases, cancer patients, transplantation, severe infections such as AIDS and severe
forms of allergy. We believe that this system would
become one of fundamental methods in clinicalimmunology.
2. Autoantibody Project
We will clarify the generative mechanism of regulatory T cells generation and will innovate the methods to control the regulatory T cells.
3. Oral tolerance Project
Our purpose is to treat autoimmune diseases using
the mechanism of tolerance induction by mucosal
immune system.
4. Intracellular Signaling Project
Identification of the ‘raft target’ is unequivocallyimportant for basic science and for therapeutic intervention of inflammatory diseases.
The pathological examination of end-stage renal
disease shows similar glomerulosclerotic patterns,although there are many causes of renal diseases.
Therefore, we believe that preventing glomerulosclerosis could stop the progression to end-stage renal disease.
5. Genetics Project
Genes wide screen of several diseases would be
expected.

Allergy
1. Airway Inflammation Project
We are expecting to establish a new strategy forpreventing the development of AHR before onset of
airway inflammation.
2. IgE Response Project
The aim of our study is to understand the pathogenesis and establish the new therapeutic strategy of
IgE-mediated allergic diseases, such as bronchial asthma and allergic rhinitis, from which more people aresuffering. Because IgE-mediated allergy seems much
more complicated than previously thought, our future
progress in this research area will be of substantial
importance.

Research Grants
1. Research on Specific Diseases from the Ministry of
Health and Welfare of Japan 1999-2001.
2. Research on Allergic disease and Immunology from
the Ministry of Health, Labour and Welfare 20022004
3. Research on Health Sciences focusing on Drug
Innovation from the Japan Health Sciences
Foundation 2000-2002
4. Research on Eye and Ear Sciences, Immunology,
Allergy and Organ Transplantation from the
Ministry of Health and Welfare of Japan 1998-2000.
5. Research Grant from the Ministry of Education,
Science, Sports and Culture of Japan(Kibann
KenkyuA) 1999-2001

Select Publications
1. Kawahata K, Misaki Y, Yamaguchi M, Tsunekawa S,
Setoguchi K, Miyazaki J and Yamamoto K. Peripheral
tolerance to a nuclear autoantigen:dendritic cells
expressing a nuclear autoantigen lead to persistentanergic state of CD4 autoreactive T cells after proliferation. J Immunol.168:1103-1112, 2002.
2. Nagase H, Miyamasu M, Yamaguchi M, Imanishi M,
Tsuno NH, Matsushima K, Yamamoto K, Morita Y
and Hirai K. Cytokine-mediated regulation of CXCR4
expression in human neutrophils. J Leukocyte
Biol.71:711-717, 2002.

3. Yoshimura C, Yamaguchi M, Iikura M, Izumi S, Kudo
K, Nagase H, Ishii A, Wall AF, Ra C, Iwata T, Igarashi
T, Yamamoto K and Hirai K. Activation makers ofhuman basophils:CD69 expression is strongly andpreferentially induced by IL-3. J Allergy Clin
Immunol.109:817-823, 2002.
4. Kawahata K, Misaki Y, Yamauchi M, Tsunekawa S,
Setoguchi K, Miyazaki J and Yamamoto K.
Generation of CD4+CD25+ regulatory T cells fromautoreactive T cells simultaneously with their negative selection in the thymus and from non-autoreactive T cells by endogenous T cell receptor expression. J Immunol. 68:4399-405, 2002.
5. Kono H, Suzuki T, Yamamoto K, Okada M, Yamamoto
T and Honda Z. Spatial raft coalescence represents
an initial step in Fc gamma R signaling. J
Immunol.169:193-203, 2002.
6. To Y, Dohi M, Matsumoto K, Tanaka R, Sato A,
Nakagome K, Nakamura T and Yamamoto K. A two
way interaction between hepatocyte growth factor
and interleukin-6 on tissue invasion of lung cancer
cell line. Am J Respir Cell Mol Biol.27:220-226, 2002.
7. Takami K, Takuwa N, Okazaki H, Kobayashi M,
Ohtoshi T, Kwasaki S, Dohi M, Yamamoto K,
Nakamura T, Tanaka M, Nakahara Y and Takizawa
H. Interferon-gamma inhibits hepatocyte growth factor-stimulated cell proliferation of human bronchial
epithelial cells.Upregulation of p27 (kip1) cyclindependent kinase inhibitor. Am J Respir Cell Mol
Biol.26:231-238, 2002.
8. Misaki Y, Ezaki I, Ariga T, Kawamura N, Sakiyama Y
and Yamamoto K. Gene-transferred oligoclonal T
cells predominantly persist in peripheral blood froman adenosine deaminase- deficinent patient during
gene therapy. Mol Ther.3:24-27, 2001.
9. Yamaguchi M, Hirai K, Komiya A, Miyamasu M,
FukumotoY, Teshima R, Ohta K, Morita Y, Galli SJ,
Ra C and Yamamoto K. Regulation of mouse mastcell surface F e RI expression by dexamethasone. Int
Immunol.13:843-851, 2001.
10. Yoshimura C, Miyamasu M, Nagase H, Iikura M,
Yamaguchi M, Kawanamai O, Morita Y, Iwata T,
Yamamoto K and Hirai K. Glucocorticoids induce
basophil apoptosis. J Allergy Clin Immunol.108:201220, 2001.
11. Yamada R, Tanaka T, Unoki K, Nagai T, Sawada T,
Ohnishi Y, Tsunoda T, Yukioka M, Maeda A, Suzuki
K, Tateishi H, Ochi T, Nakamura Y and Yamamoto
K. Association between a single-nucleotide polymorphism in the promoter of the human interleukin-3
gene and rheumatoid arthritis in Japanese patients,and maximum-likelihood estimation of combinatorial effect that two genetic loci have on susceptibility
to the disease. Am J Hum Genet.68:674-685, 2001.
12. Kawasaki S, Takizawa H, Yoneyama H, Nakayama T,
Fujisawa R, Izumizaki M, Imai T, Yoshie O,
Hommma I, Yamamoto K and Matsushima K.
Intervention of thymus and activation-regulated
chemokine attenuates the development of allergic

179airway inflammation and hyperrespnsiveness inmice. J Immunol.166:2055-2062, 2001.
13. Setoguchi K, Misaki Y, Terauchi Y, Yamauchi T,
Kawahata K, Kadowaki K and Yamamoto K.
Peroxisome proliferator-activated receptor-γ haploinsufficiency enhances B cell proliferative responses and exacerbates experimentally induced arthritis.
J Clin Invest.108:1667-1675, 2001.
14. To Y, Dohi M, Tanaka R., Sato A, Nakagone K, andYamamoto K. Early IL-4-dependent response can
induce airway hyperreactivity in amouse model ofasthma. Lab Invest.81:1385-96, 2001.
15. Kanda H, Kubo K, Hamasaki K, Kanda Y, Nakao A,
Kitamura T, Fujita T, Yamamoto K, Mimura T.
Influence of various hemodialysis membrane on theplasma (1→3)-β-D-glucan level. Kidney Int.60:31923, 2001.
16. Nagase H, Miyamasu M, Yamaguchi M, Fujisawa T,
Ohta K, Yamamoto K, Morita Y and Hirai K.
Expression of CXCR4 in eosinophils:Functional
analyses and cytokine- mediated regulation. J
Immunol.164:5935-43, 2000.
17. Sasaki M, Nakamura S, Ohyama Y, Shinohara M,
Ezaki I, Hara H, Kadena T, Kishihara K, Yamamoto
K, Nomoto K and Shirasuna K. Accumulation of common Tcell clonotypes in the salivary glands ofpatients with human T lymphotropic virus typeIassociated and idiopathic Sjögren’s syndrome. J
Immunol.164:2823-2831, 2000.
18. Fujio K, Nosaka T, Kojima T, Kawashima T, Yahata
T, Copeland NG., Gilbert DJ, Jenkins NA, Yamamoto
K, Nishimura T and Kitamura T. Molecular cloning of
a novel type 1 cytokine receptor similar to the common gamma chain. Blood.95:2204-2210, 2000.
19. Sawabe T, Shiokawa S, Sugisaki K, Tsuda T and
Yamamoto K. Accumulation of common clonal T
cells in multiple lesions of sarcoidosis. Mol
Medicine.6:793-802, 2000.
20. Sekiya T, Miyamasu M, Imanishi M, Yamada H,
Nakajima T, Yamaguchi M, Fujisawa T, Pawankar R,
Sano Y, Ohta K, Ishii A, Morita Y and Yamamoto K.
Matsushi K. Yoshie O. and Hirai K. Inducible expression of a Th2-type CC chemokine thymus- and activation-regulated chemokine (TARC) by humanbronchial epithelial cells. J Immunol.165:2205-2213,
2000.
21. Nagase H, Yamaguchi M, Jibiki S, Yamada H, Ohta K,
Kawasaki H, Yoshie O, Yamamoto K, Morita Y andHirai K. A rapid and simple photometric assay forquantification of eosinophil chemotaxis. Int Arch
Allergy Immunol.122:10-14, 2000.
22. Fujio K, Misaki Y, Setoguchi K, Morita S, Kawahata
K, Kato I, Nosaka T, Yamamoto K and Kitamura T.
Functional reconstitution of classII MHC-restricted
Tcell immunity mediated by retroviral transfer of thea b TCR complex. J Immunol.165:528- 532, 2000.
23. Setoguchi K, Misaki Y, Araki Y, Fujio K, Kawahata K,
Kitamura T and Yamamoto K. Antigen-specific T
cells transduced with IL-10 ameliorate experimentally induced arthritis without impairing the systemic

180

immune response to the antigen. J Immunol.
165:5980-5986, 2000.
24. Nagase H, Miyamasu M, Yamaguchi T, Kawasaki H,
Ohta K, Yamamoto K, Morita Y and Hirai K.
Glucocorticoids preferentially upregulate functional
CXCR4 expression in eosinophils. J Allergy Clin
Immunol.106:1132-1139, 2000.
25. Suzuki T, Kono H, Hirose N, Okada M, Yamamoto T,
Yamamoto K and Honda Z. Differential involvementof Src family kinases in Fc gamma receptor-mediatedphagocytosis. J Immunol.165:473-482, 2000.
26. Honda Z, Suzuki T, Kono H, Okada M, Yamamoto T,
Ra C, Morita Y and Yamamoto K. Sequential requirements of the N-terminal palmitoylation site and SH2
domain of Src family kinases in the initiation andprogression of Fcepsilon RI signaling. Mol Cell
Biol.20:1759-1771, 2000.
27. Furukawa H, Yabe T, Watanabe K, Miyamoto R, Miki
A, Akaza T, Tadokoro K, Tohma S, Inoue T,
Yamamoto K and Juji T. Tolerance of NK and LAK
activity for HLA class I-deficient targets in a TAPdeficient patient. (bare lymphocyte syndrome type I)
Hum Immunol.60:32-40, 1999.
28. Matsui T, Kurokawa M, Kobata T, Oki S, Azuma M,
Tohma S, Inoue T, Yamamoto K, Nishioka K and
Kato T. Autoantibodies to Tcell co-stimulatory molecules in systemic autoimmune diseases. J
Immunol.162:4328-4335, 1999.
29. Furukawa H, Murata S, Yabe T, Shimbara N, Keicho
N, Kashiwase K, Watanabe K, Ishikawa Y, Akaza T,
Tadokoro K, Tohma S, Inoue T, Tokunaga K,
Yamamoto K, Tanaka K and Juji T. Splice acceptor
site mutation of transporter associated with antigen
processig-1 gene in human bare lymphocyte syndrome. J Clin Invest.103:755-758,1999.
30. Nagahori T, Dohi M, Matsumoto K, Saitoh K, Honda
ZI, Nakamura T and Yamamoto K. Interferon-gamma
upregulates the c-Met/hepatocyte growth factor
receptor expression in alveolar epithelial cells. Am J
Resp Cell Mol Biol.21:490-497, 1999.
31. Kawahata K, Misaki Y, Komagata Y, Setoguchi K,
Tsunekawa S, Yoshikawa Y, Miyazaki J andYamamoto K. Altered expression level of a systemic
nuclear autoantigen determines the fate of immune
responce to self. J Immunol.162:6482-6491, 1999.
32. Furukawa H, Yabe T, Akaza T, Tadokoro K, Tohma
S, Inoue T, Tokunaga K, Yamamoto K, Geraghty DE
and Juji T. Cell surface expression of HLA-E molecules on PBMC from a TAP1-deficient patient. Tissue
Antigens.53:292-295, 1999.
33. Takizawa H, Ohtoshi T, Kawasaki S, Kohyama T,
Desaki M, Kasama T, Kobayashi K, Nakahara K,
Yamamoto K, Matsushima K and Kudoh S. Diesel
exhaust particles induce NF-κB activation in humanbronchial epithelial cells in vitro:Importance incytokine transcription. J Immunol.162:47054711,1999.
34. Miyamasu M, Yamaguchi M, Nakajima T, Misaki Y,
Morita Y, Matsushima K, Yamamoto K and Hirai K.
Th1-derived cytokine IFN-γ is a potent inhibitor ofeotaxin sythesis in vitro. Int Immunol.11:1001-1004,
1999.
35. Iikura M, Yamaguchi M, Miyamasu M, Morita Y,
Iwase T, Moro I, Yamamoto K and Hirai K. Secretory
IgA-mediated basophil activation. Biochem BiophRes Co.264:575- 579, 1999.
36. Miyamasu M, Nakajima T, Misaki Y, Izumi S, Tsuno
N, Kasahara T, Yamamoto K, Morita Y and Hirai K.
Dermal fibroblasts represent a potent major sourceof human eotaxin:in vitro production and cytokinemediated regulation. Cytokine.11:751-758, 1999.
37. Murata S, Kawahara H, Tohma S, Yamamoto K,
Kasahara M, Nabeshima Y, Tanaka K and Chiba T.
Growth retardation in mice lacking the proteasome
activator PA28g. J Biol Chem.274:38211-38215, 1999.
38. Kanda H, Mimura T, Hamasaki K, Yamamoto K,
Yazaki Y, Hirai H and Nojima Y. Fyn and Lck tyroine
kinase regulate tysine phoshorylation of p105CasL, amember of the p130Cas docking protein family,in Tcell receptor-mediated signalling. Immunol. 97:56-61,
1999.
39. Kubo K, Miyagawa K, Yamamoto R, Hamasaki K,
Kanda K, Fujita T, Yamamoto K, and Mimura T.
Detection of WT1 mRNA in urine from Patients withkidney diseases. Eur J Clin Invest.29:824-826, 1999.
40. Dohi M, Tsukamoto S, Nagahori T, Shinagawa K,
Saitoh K, Tanaka Y, Kobayashi S, Tanaka R, To andYamamoto K. Noninvasive system for evaluating theallergen- pecific airway response in a murine modelof asthma. Lab Invest.79:1559-15, 1999.
41. Furukawa H, Yabe T, Watanabe K, Miyamoto R,
Akaza T, Tadokoro K, Tohma S, Inoue T, Yamamoto
K and Juji T. An alternatively spliced form of thehuman CD94 gene. mmunogenetics.48:87-88, 1998.
42. Nakajima T, Yamada H, Iikura M, Miyamasu M,
Izumi S, Shida H, Ohta K, Imai T, Osamu Y,
Mochizuki M, Schroder J, Morita Y, Yamamoto K
and Hirai K. Intracellular localization and release of
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Transcriptional roles of C/EBP-β, NF-κB,and CBF/minterleukin-Iβ-induced interleukin-6 synthesis byhuman rheumatoid fibroblast-line synoviocytes. J
Biol Chem.273:7620-7627, 1998.
44. Miyazawa K, Mori A, Yamamoto K and Okudaira H.
Constitutive transcription of the human IL-6 gene byrheumatoid synoviocytes spontaneous activation of
NF-κB and CBFI. Am J Pathol.152:793-803, 1998.
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YamamotoK. Establishment of a novel B cell clonality analysis using single-strand conformation polymorphism of immunoglobulin light chain messenger
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Yamamoto T and Honda Z. Essential roles of Lyn infibronectin-mediated filamentous actin assembly andcell motility in mast cells. J Immunol.161:3694-3701,
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47. Iikura M, Yamaguchi M, Fujisawa T, Miyamasu M,
Takaishi T, Morita Y, Iwase T, Moro I, Yamamoto K
and Hirai K. Secretory IgA induces degranulationfrom IL-3- primed basophils. J Immunol.161:15101515, 1998.
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Tashiro F, Komagata Y, Maki K, Ikuta K, Ouchi Y,
Miyazaki J, and Shimizu T. Role of cytosolic phospholipase A2 in allergic response and parturition.
Nature.390:618-622, 1997.
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Department of
Psychosomatic Medicine
Outline and Research Objectives
Department of Psychosomatic Medicine was founded in 1972 at the Branch Hospital of
the University of Tokyo.
Our department started as a clinical section for seeing ‘psychosomatic diseases’ that arephysical diseases whose onset and courses are greatly affected by psychosocial, stress-related factors in the field of internal medicine. However, we have come to cover many patientsof eating disorders, panic disorders, and mild depression that are not strictly psychosomatic
diseases but share common features of the presence of many physical symptoms and theinfluence of psychosocial factors.
Based on these historical circumstances, our department has mainly focused on the
problems of eating disorders and has also studied panic disorder and mild depression.
However, we have also actively studied lifestyle-related and adult diseases such as hypertension (HT), diabetes mellitus (DM), Basedow’s disease, and cancers in order to meet thesocial needs expecting us to see stress-related diseases in the field of internal medicine,investigating what kinds of psychosocial factors are related to their onset and courses andwhat kinds of psychobehavioral interventions are effective on these diseases. Moreover, wehave performed basic and interventional studies of various relaxation procedures that could
cancel hazardous effects of stress on our health.
Because psychosomatic medicine on which we rely is rather a new field of medical science born about half a century ago and thus special research methodologies have not been
established yet, we should have developed a new method for performing new research.
Moreover, the pathological mechanisms of our subjects such as psychosomatic diseases andeating disorders are so complicated by many factors including physical, psychological andbehavioral ones, and it has been very difficult for us to empirically uncover their roles one
by one.
However, the conditions surrounding our field of research have dramatically changed inrecent years, because many new kinds of methods have been developed that can be applied
to psychosomatic field including brain functional imaging procedures, ecological momentary
assessment (EMA) which means successive monitoring of biopsychobehavioral time series
data in daily life, and the discovery and development of basic research of novel eating-related substances. Furthermore, the increase of various psychosocial stress and the acknowledgment of its hazardous effects on health in modern world heighten the social needs forthe evidence-based interventional procedures of stress-related diseases far stronger than
before. Therefore, our department must greatly contribute to our society in the near future.

Faculties and Students
Professor and Chair

Tomifusa Kuboki, MD,PhD, Since
April 1996
Associate Professors Hiroaki Kumano, MD,PhD
Associate .................................2
Graduate student ....................7
Research student.....................1
Secretary .................................3

Past Research and Major Accomplishments
We have mainly performed clinical research ofpatients consulting our department and behavioral

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scientific research of life-related and adult diseases
consulting the department of internal medicine andother physical departments. Furthermore, as a basic
research, we have investigated one of the main subjects of psychosomatic medicine that is mind/bodycorrelation meaning interaction between biological,
psychological, and behavioral aspects of human being
through the study of stress and relaxation. The main
fields of our research are as follows and related papers
are listed in parentheses the number of which corresponds to the list of Selected Publicatios.
1) Clinical, behavioral scientific, and epidemiologicalstudiesoneatingdisorders(#1,2,3,4,5,10,11,14,16,23,25,30)

2) Behavioral scientific, psychophysiological, andinterventional studies on lifestyle-related andadult diseases including DM, HT, Basedow’s disease,andcancers(#9,13,17,18,21,22,24,26,28,29,31,33,34)
3) Studies on other clinical subjects of our department
including panic disorder and depression(#6,12,20,27,32,36,37)
4) Studies on the development of questionnaires concerning mental health including the TEG, the TSS,
the POMS, the LHQ, the SIRI33, and the SSA(#7,28,31,33,35)
5) Psychophysiological and interventional studies on
various relaxation procedures such as autogenic
training, biofeedback, photic feedback, and Qigong (#8,15,18,19,26,29,34)
Concerning eating disorders, we did the first country-wide epidemiological study and made the mannual of cognitive bahavioral therapy and family guidancefor carrying out controlled treatment study collaborating on the projects of Ministry of Health and Welfare.
Furthermore, we did many clinical studies for clarifying various features of these patients.
Concerning lifestyle-related and adult diseases, wehave introduced advanced methodologies of behaviralsciences and data analysis prodedures including various multivariate analyses. We have gathered many
data on how psychobehavioral problems such as
lifestyles, stress, and personality could affect theonset and courses of these diseases. These data areuseful for clarifying how we should change what kindof behavior for behavior modification that are regarded to be important for the treatment of these diseases.
Stress and mental health are important research
themes of psychosomatic medicine, and we investigated hazardous effects of stress by developing various psychobehavioral questionnaires. We got excellent results such as developing the Tokyo University
Egogram (TEG) that are now widely used not only in
hospitals but also in companies and schools all overJapan, and the Life Health Questinnaire (LHQ) for
assessing multiple stress-related variables based on
the data of more than 10,000 healthy people.
Furthermore, the questionnaire for examining the riskof certain personality trait on the occurrence of cancers named the Short Interpersonal Reactions
Inventory (SIRI33) was also developed, and a collaborative study with the National Cancer Center is now
in progress.
Because we speculate that the cumulative effects ofstress may precipitate or aggravate psychosomatic diseases, panic disorder, or depression, it is urgently necessary to develop therapeutic procedures soothing theeffects of stress. We have investigated various relax-

ation procedures as such stress-reducing methods. As
basic research, physical and psychological characteristics of relaxation response have been studied throughpsychophysioloical measures of autonomic nervous
system and electroencephalogram (EEG), and a questionnaire assessing psychological relaxation response
has been developed. Furthermore, we have done several interventional studies targeting DM, psyochosomatic diseases, or depression, and excellent results
have been obtained such as very positive effects of Qigong group therapy on metabolic control of type 2
DM substantiated by a randomized controlled trial(RCT).
Because psychosomatic medicine is rather a new
field of medical science, we should have developed
new methods or introduced various methods fromadjacent fields for performing new research. As mentioned above, we have achieved several positive
results in this point and have made it possible to gather and analyse relevant data, which had never beenpossible before.

Current Research
We are now advancing more basic research investigating the relationship between physical, psychological and behavioral aspects in the onset and courses ofvarious stress-related diseases using newly developed
research methodologies such as noninvasive assessment procedures of brain or autonomic nervous system function and EMA of human time-series bioinformation. Furthermore, it should be noted that targets of research have been expanded to include subjects complaining mainly fatigue who suffer fromchronic fatigue syndrome (CFS) and multiple chemicalsensitvities (MCS) and both healthy and mildly
demented geriatric poplulation.
Now we have four active research laboratories inour department, and each is pursuing various research
projects as follows making full use of unique methodologies and collaborating with many outside institutes
and researchers.
1) Behavioral Science Laboratory
Objectives: In order to clarify the disease processes and ways of treatment of lifestyle-related diseases
and psychosomatic diseases, we should deal with thefunctional aspect of subjects fucusing on behavior
meaning how they live a daily life as well as theanatomical aspect focusing on body and the introspective aspect focusing on mind. The objectives of thislaboratory are to investigate behavior as well as its
relation with mind and body of various subjects ranging from psychosomatic patients to healty persons.
Methodologies: Using questionnaires, personal
degital assistants, psychophysiological assessment

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procedures, or blood chemistry, the multiple data concerning behavior, mind and body are gathered. Then,the structure within the data is precisely extracted bymaking full use of many kinds of data analysis methods such as multivariate analyses, time-series analyses, and analysis of variance.
Projects:
DM: Cross-sectional and longitudinal studies toclarify multiple psychosocial factors affecting metabolic control and quality of life (QOL) of diabetic patients
Tension-type headache: An interventional study
by autogenic training
Cancer: A semi-prospective study of the role oftype C personality on the onset of prostate cancer; A
prospective study of personality factors affecting theonset of various cancers in healthy persons
Renal failure: A cross-sectional study on the relatonship between stress-related factors and metabolic
control and QOL of hemodialysis patients
MCS: A cross-sectional study on stress-related factors affecting their onset and courses
Depression: To determine the diagnostic criteria ofmild depression
Elderly people: Cross-sectional and longitudinalstudies to clarify the relationship between bodilyfunction, exercise ability, lifestyle, and stress-related
factors2) Bio-signal Analysis Laboratory
Objectives: Although human body produces various kinds of signals, most of them are not captured
and discarded as noise. However, as many novel signal analysis methods have been developed these days,
we can now extract valuable information from signals
previously regarded as noise and can investigate sucha field as has been difficult to be targeted by scientificinquiries. The objectives of this laboratory are tounderstand pathophysiology, mind/body correlation,
stress, or relaxation by using such novel methodologies.
Methodologies: Higher brain functions are analyzed by use of positron emission tomography (PET),
functional magnetic resonance imaging (fMRI), magnetic encephalography (MEG), and EEG. Autonomic aswell as central nervous functions are estimated byspectral and fractal analyses of heart rate variabilities.
Ecological changes of heart rate variabilities, bodily
movement, mood, symptoms, and toxic substances inthe environment are successively monitored by EMA
methods.
Projects:
PET: Measurement of cerebral blood flow or glucose metabolism before and after the treatment ofanorexia nervosa or panic disorders; Investigation on
the relationship between stress coping processes andcerebral blood flow

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EEG: Studies assessing the improvement of cerebral cortical function by some relaxation procedures
using the dipole tracing method; Assessment ofpatients with various diseases by event-related potentials (ERP)
Heart rate variabilities & EMA: A study on autonomic nervous activities and bodily movement in therefeeding phase of anorexia nervosa; Development ofautonomic nervous function database of outpatientsin our department; Comprehensive assessment ofmultiple chemical sensitivities and chronic fatigue
syndrome by EMA methods
3) Neuro-immuno-endocrinology Laboratory
Objectives: The biological basis of mind/body correlation is regarded as ‘functional association between
neuroendocrine and immune systems’, that is sought
to be clarified by clinical as well as basic methods inthis laboratory. Brain mechanism affecting abnormal
eating behavior is also a main focus to be inquired. In
addition, the search for physiological measures ofstress responses and their application to clinical field
is another challenge.
Methodologies: Methods of functional morphology and behavioral analysis are used for analyzing
genetically-modified mice such as a knockout mouse.
We take notice of 8-hydroxy-2’-deoxy guanosine(8OHdG) as an oxidated substance of DNA and measure it in human urine.
Projects:
Inflammatory cytokines: Analyses of expression
and mechanism of action in brain tissue of inflammatory cytokines such as IL-1 and IL-6 as modulators ofneuroendocrine and immune systems
Eating modulating substances: Investigation on
the effects of newly-found neuropeptides such as
orexin and ghrelin in brain on emotion and stressresponses
Novel stress measure: Investigation on the usefulness of 8OHdG as a physiological measure in humanstress-related diseases
4) Eating Disorders Laboratory
Objectives: The etiology of eating disorders suchas anorexia nervosa and bulimia nervosa wasassumed to be affected by many factors including
individual psychological, biological, familial, socio-cultural factors. The objectives of this laboratory are to
study various factors relating to the courses of eating
disorders, to develop and improve treatment procedures, and to do the basic research of pathophysiology collaborating with other laboratories.
Methodologies: Bio-psycho-behavioral clinicaldata at the initial visit and during treatment courses
are gathered; Pathophysiology of eating disorders isinvestigated by PET or EMA.

Projects:
Dropout: A study by survival analysis on premature termination of outpatient treatment
Refeeding: Assessment of energy metabolism,
autonomic nervous function, and bodily movement in
the refeeding phase of anorexia nervosa patients
Cerebral blood flow and dopaminergic metabolism:
Pet studies of anorexia nervosa patients

Future Prospects
We want to continue to advance activities ofabove-mentioned four laboratories and to focus on thefollowing points in particular.
1) Brain processes closely related to mind/body correlation in various stress-related diseases and pathogenic brain functional abnormalities will be clarified by active use of functional brain analysis
methods such as PET, fMRI, MEG, ERP, and dipole
tracing methods.
2) Ecological changes in everyday life of biological,
psychological, behavioral, and environmental
measures will be monitored and analyzed in orderto characterize neurobehavioral basis of various
stress-related diseases by developing and utilizing
relevant EMA methods.
3) Treatment programs will be developed that intervene psychobehavioral factors found to be closely
related to the processes of psychosomatic diseases
and lifestyle-related diseases by our behavioral scientific studies and include effective relaxation procedures. Then, the evidence will be shown thatthese interventions are effective in improving not
only symptoms but also QOL and life satisfaction
by carrying out RCT of these treatment programs.
4) Biological and material basis of eating disorders will
be pursued by both basic research such as one
using genetically-modified mice and clinicalresearch measuring eating-related substances inthe blood of eating disorders patients before andafter oral glucose tolerance tests.
In summary, we are going to advance toward two
directions. First, we will change over from the previous research system mainly relying on clinical andpsychobehavioral domains to the new research system that enables us to investigate neurobehavioral
and biological mechanisms actively utilizing novel
methodologies. Then, we will accumulate useful data
for the development of biological treatment by broadening our horizons from mind/body correlation to
mind/brain/body correlation and by seeking for thetrue nature of pathological processes of stress-relateddiseases.
Secondly, we will make it possible to gather
diverse data in patients’ everyday life by incorporating

EMA into behavioral scientific methodologies, andthen we will aim to clarify the reality of not only diseases themselves but also patients having such diseases. Based on these results as well as our present
knowledge, we will develop treatment programs
including interventions manipulating psychobehavioral variables and relaxation procedures and will clarify by RCT what kinds of interventions most effectively improve not only symptoms but also QOL andlife satisfaction.

Research Grant
1) Grant of Ministry of Health, Labor and Welfare Sick
House Syndrome(SHS)---Studies of patho-physiology,
method of diagnosis and its treatment.
2) Grant of Ministry of Health, Labor and Welfare Eating
disorder- Studies of neurology and endocrinology.
3) Grant of Ministry of Health, Labor and Welfare A
guideline for the diagnosis and treatment of psychosomatic diseases 2002
4) Grant of Ministry of Health and Welfare Studies ofstress management

Select Publications
1. Suematsu H, Ishikawa H, Kuboki T, Ito T. Statisticalstudies on anorexia nervosa in Japan: detailed clinicaldata on 1,011 patients. Psychother Psychosom43(2):96-103, 1985.
2. Suematsu H, Kuboki T, Itoh T. Statistical studies on
the prognosis of anorexia nervosa. Psychother
Psychosom 43(2):104-12, 1985.
3. Suematsu H, Kuboki T, Ogata E. Anorexia nervosa inmonozygotic twins. Psychother Psychosom 45(1):4650, 1986.
4. Kuboki T, Suematsu H, Ogata E, Yamamoto M,
Shizume K. Two cases of anorexia nervosa associatedwith Graves’ disease. Endocrinol Jpn 34(1):9-12, 1987.
5. Kumano H, Kuboki T, Tawara R, Sasaki T, Nomura S,
Suematsu H, Ogata E. Interrelationship between
serum muscle enzymes and a low T3 in anorexia nervosa. Endocrinol Jpn 37(4):583-9, 1990.
6. Kuboki T, Ide M, Kumano H, Nomura S, Suematsu H.
Historical review of panic disorder and clinical studies
in Japan． Journal of Psychiatric Research24(Suppl.1):70-72, 1990.
7. Kuboki T, Nomura S, Wada M, Akabayashi A,
Nagataki M, Suematsu H, Yokoyama K, Araki S.
Multidimensional assessment of mental state in occupational health care--combined application of threequestionnaires: Tokyo University Egogram (TEG),
Time Structuring Scale (TSS), and Profile of Mood
States (POMS). Environ Res 61(2):285-98, 1993.
8. Nakamoto C, Kumano H, Horie H, Nomura S, Kuboki
T, Suematsu H, Yasushi M, Saitou S, Chijiiwa M.
Psychological and physiological effects of photic feedback system. Shinshin-Igaku 33(5):371-80, 1993.

185

9. Horie H, Kumano H, Nomura S, Kuboki T, Suematsu
H, Hagura R. Multiple psychosocial factors influencing diabetes mellites: Multivariate analysis by quantification type I method. Shinshin-Igaku 33(8):667674, 1993.
10. Berger D, Saito S, Ono Y, Tezuka I, Shirahase J,
Kuboki T, Suematsu H. Dissociation and child abuse
histories in an eating disorder cohort in Japan. Acta
Psychiatr Scand 90(4):274-80, 1994.
11. Berger D, Ono Y, Saito S, Tezuka I, Takahashi Y, Uno
M, Ishikawa Y, Kuboki T, Asai M, Suematsu H.
Relationship of parental bonding to child abuse anddissociation in eating disorders in Japan. Acta
Psychiatr Scand 91(4):278-82, 1995.
12. Kumano H, Kuboki T, Ide M, Okabe F, Suematsu H.
Exercise tolerance evaluated by respiratory gas analysis during treadmill exercise test in panic disorderpatients．Jpn J Psychosom Med 35(8):671-679, 1995.
13. Horie H, Kumano H, Kuboki T, Suematsu H, Hagura
R. Multiple psychosocial factors influencing noninsulin-dependent diabetes mellites (II). ShinshinIgaku 35(6):501-509, 1995.
14. Kuboki T, Nomura S, Ide M, Suematsu H, Araki S.
Epidemiological data on anorexia nervosa in Japan.
Psychiatry Res 16;62(1):11-6, 1996.
15. Kumano H, Horie H, Shidara T, Kuboki T, Suematsu
H, Yasushi M. Treatment of a depressive disorderpatient with EEG-driven photic stimulation.
Biofeedback Self Regul 21(4):323-34, 1996.
16. Kumano H, Yamauchi Y, Matsumoto S, Sakano Y,
Kuboki T, Suematsu H. Cognitive behavior therapy
for eating disorders: Its advantages and shortcomings. Jpn J Psychosom Med 37(1):55-60, 1997.
17. Yoshiuchi K, Nomura S, Ando K, Ohtake T,
Shimosawa T, Kumano H, Kuboki T, Suematsu H,
Fujita T. Hemodynamic and endocrine responsiveness to mental arithmetic task and mirror drawing
test in patients with essential hypertension. Am J
Hypertens 10(3):243-9, 1997.
18. Nakao M, Nomura S, Shimosawa T, Yoshiuchi K,
Kumano H, Kuboki T, Suematsu H, Fujita T. Clinicaleffects of blood pressure biofeedback treatment on
hypertension by auto-shaping. Psychosom Med
59(3):331-8, 1997.
19. Kumano H, Horie H, Kuboki T, Suematsu H, Sato H,
Yasushi M, Kamei T, Masumura S. EEG-driven photic
stimulation effect on plasma cortisol and beta-endorphin. Appl Psychophysiol Biofeedback 22(3):193-208,
1997.
20. Nakao M, Nomura S, Yamanaka G, Kumano H,
Kuboki T. Assessment of patients by DSM-III-R and
DSM-IV in a Japanese psychosomatic clinic.
Psychother Psychosom 67(1):43-9, 1998.
21. Nakao M, Shimosawa T, Nomura S, Kuboki T, Fujita
T, Murata K. Mental arithmetic is a useful diagnostic
evaluation in white coat hypertension. Am J
Hypertens 11(1 Pt 1):41-5, 1998.
22. Yoshiuchi K, Kumano H, Nomura S, Yoshimura H,
Ito K, Kanaji Y, Ohashi Y, Kuboki T, Suematsu H.
Stressful life events and smoking were associated186with Graves’ disease in women, but not in men.
Psychosom Med 60(2):182-5, 1998.
23. Nakao M, Kumano H, Nomura S, Kuboki T,
Yokoyama K, Murata K. Fatigue mood as an indicator discriminating between anorexia nervosa andbulimia nervosa. Acta Psychiatr Scand 97(3):202-5,
1998.
24. Yoshiuchi K, Kumano H, Nomura S, Yoshimura H,
Ito K, Kanaji Y, Kuboki T, Suematsu H. Psychosocial
factors influencing the short-term outcome ofantithyroid drug therapy in Graves’ disease.
Psychosom Med 60(5):592-6, 1998.
25. Nakao M, Kumano H, Nomura S, Kuboki T, Murata
K. Assessment of ego state in anorexia nervosa andbulimia nervosa. Acta Psychiatr Scand 99(5):385-7,
1999.
26. Nakao M, Nomura S, Shimosawa T, Fujita T, Kuboki
T. Blood pressure biofeedback treatment, organ damage and sympathetic activity in mild hypertension.
Psychother Psychosom 68(6):341-7, 1999.
27. Fukunishi I, Berger D, Wogan J, Kuboki T.
Alexithymic traits as predictors of difficulties withadjustment in an outpatient cohort of expatriates inTokyo. Psychol Rep 85(1):67-77, 1999.
28. Kumano H, Orii Y, Suzukamo Y, Yamauchi Y,
Munakata M, Yoshinaga K, Seto M, Sakano Y, AgariI, Kuboki T. A trial of developing the Japanese briefversion of the Short Interpersonal Reactions
Inventory: To measure the Cancer-prone Personality.
Jpn J Psychosom Med 39(5):55-60, 1999.
29. Nakao M, Nomura S, Shimosawa T, Fujita T, Kuboki
T. Blood pressure biofeedback treatment of whitecoat hypertension. J Psychosom Res 48(2):161-9,
2000.
30. Nakahara R, Yoshiuchi K, Yamanaka G, Sasaki T,
Suematsu H, Kuboki T. Coping skills in Japanesewomen with eating disorders. Psychol Rep 87(3 Pt
1):741-6, 2000.
31. Kumano H, Orii Y, Yamauchi Y, Seto M, Agari I,
Sakano Y, Munakata M, Yoshinaga K, Sasaki T,
Kuboki T. A trial of developing the Japanese briefversion of the Short Interpersonal Reactions
Inventory (2nd report): Revision to the 33 items version. Jpn J Psychosom Med 40(6):447-54, 2000.
32. Nakao M, Yamanaka G, Kuboki T. Major depression
and somatic symptoms in a mind/body medicine
clinic. Psychopathology 34(5):230-5, 2001
33. Kumano H, Kuboki T, Orii Y, Fukuse T, Hirata T,
Shinohara K, Seto M, Agari I, Sakano Y. The investigation of the discriminant validity of the measurement of type C personality using the Japanese briefversion of the Short Interpersonal Reactions
Inventory (SIRI33). Jpn J Psychosom Med 41(8):59399, 2001.
34. Tsujiuchi T, Kumano H, Yoshiuchi K, He D, Tsujiuchi
Y, Kuboki T, Suematsu H, Hirao K. The effect of Qigong relaxation exercise on the control of type 2 diabetes mellitus: a randomized controlled trial.
Diabetes Care 25(1):241-2, 2002.

35. Nakao M, Barsky AJ, Kumano H, Kuboki T.
Relationship between somatosensory amplification
and alexithymia in a Japanese psychosomatic clinic.
Psychosomatics 43(1):55-60, 2002.
36. Nakao M, Yamanaka G, Kuboki T. Suicidal ideationand somatic symptoms of patients with mind/body
distress in a Japanese psychosomatic clinic. SuicideLife Threat Behav 32(1):80-90, 2002.
37. Nakao M, Ando K, Nomura S, Kuboki T, Uehara Y,
Toyooka T, Fujita T. Depressive mood accompanies
hypercholesterolemia in young Japanese adults. Jpn
Heart J 42(6):739-48, 2001.

187

Department of ClinicalLaboratory Medicine
Outline and Research Objectives
Department of Clinical Laboratory Medicine, Tokyo University Graduate School of
Medicine, was established in 1997 following foundation of Department of Laboratory
Medicine, Faculty of Medicine, University of Tokyo in 1984. We have also ClinicalLaboratory Center which was established in 1952 in Tokyo University Hospital. The
Department cooperates with Clinical Laboratory Center not only in giving lectures and clinical education to medical students but also in carrying out researches. The Department ofClinical Laboratory Medicine is composed of only two staffs, a professor and an associate
professor, but Clinical Laboratory Center possesses four positions of lecturers and two of
associates. The professor and the associate professor of the Department are the director andthe vice-director of Clinical Laboratory Center, respectively.
In our department five or six research projects have been usually undertaken.

Faculties and Students
Professor and Chair Kazuhiko Nakahara, M.D. (1997~)
Associate Professor Kiyoshi Kitamura, M.D.(1998~2002)
Graduate Student....................1
Secretary .................................1

Past Research and Major Accomplishments
1. Cell surface analysis using flow cytometry
The major purpose of this project was to apply the
immunological methods to actual clinical fields, forinstance to diagnosis hematological malignancies suchas leukemia and lymphoma. We found that a kind of
glycolipids, asialo GM1, is the candidate of cell surface
markers for diagnosis of acute lymphoblastic
leukemia by using the method of flow cytometry. For
the next steps in this project, we discovered flow
cytometric analysis using several monoclonal antibodies reacted against human blood cells, especially lymphocytes, is very useful tool to diagnose humanhematological disorders. To give an example, we mentioned that adult T-cell leukemia cells are CD4 positive but negative for CD8, and very clear pattern ofthe positive sharp peak could be shown on the display of flow cytometry. We also demonstrated that
not only single-color but also multi-color analysis
with monoclonal antibodies and flow cytometry isvery important to investigate some kinds of illness inmore detail.
For the purpose of evaluating lymphocyte activation, we have studied lymphocyte surface antigens,plasma soluble CD4 or CD8, and intracellular pH (pHi)of lymphocytes, especially in infectious mononucleo-

188

sis (IM) in which proliferation of T lymphocytes is
supposed to play an important role in regulation of EB
virus. Immunofluorescence analysis demonstrates
increase or decrease of lymphocyte subset as well as
expression of activation antigens. During IM, CD8+ T
cells are quantitatively the major population of
expanded lymphocytes and they coexpress HLA-DR
and CD45RO. Elevated serum levels of soluble CD8(sCD8) and soluble CD4 (sCD4) are regarded as markers of T cell activation. We demonstrated marked
increase in sCD8 and significant increase in sCD4
which strongly suggest activation of both CD8+ andCD4+. The elevation of plasma sCD8 is due to expansion of CD8+ subset as well as increased sCD8 releasefrom each CD8+ cell.
Additionally we tried to apply the flow-cytometricanalysis method to measure intracellular pH (pHi),
which shows one of the indices of cellular activation.
For the purpose of evaluating intracellular pH of eachlymphocyte subset, simultaneous measurement of
pHi and cell surface antigen was developed using 2’-7’bis(carboxyethyl)5,6-carboxyfluorescein (BCECF) andflow-cytometry. A significant increase in pHi of CD8+lymphocytes from patients with IM and its correlation with number of CD8+-HLA-DR+ lymphocytes aswell as plasma levels of soluble CD8 strongly suggest
activation of CD8+ lymphocytes.
2. Effects of age, body-mass index and life-style
habits on clinical laboratory data.
2-1 Alcohol drinking and clinical laboratory data
Epidemiological studies have consistently shownthat light or light/moderate drinkers are at a lower
risk of coronary heart disease (CHD). The mecha-

nisms of this association include beneficial effects on
HDL- and LDL-cholesterol, insulin sensitivity, platelet
aggregation, blood coagulation and fibrinolysis.
However, drinking also has disadvantageous effects
on blood pressure, triglycerides and uric acid.
The purpose of our study was 1) to clarify whether
there are individual varieties regarding the responseof clinical laboratory data to alcohol consumption,and 2) what are the factors responsible for the individual varieties.
2-1-1 Are there individual varieties regarding the
response of clinical laboratory data to alcohol
consumption?
We performed a cross-sectional study of 3130
Japanese male workers with a body-mass index below
24. The subjects were divided into 2 groups; a normal
r-glutamyl transpeptidase (rGTP) (<40 IU/L) group anda high rGTP (>40 IU/L) group, and the values werecompared after adjusted for age, body-mass index,
exercise and smoking. The level of triglycerides
increased according to the amount of drinking in thehigh rGTP group, whereas no association wasobserved in the normal rGTP group. The level of HDLcholesterol increased with drinking in the normal andhigh rGTP groups, and no difference was observed in
the levels of HDL-cholesterol between the twogroups. The levels of uric acid and blood pressure also
increased with drinking in both groups, but theincrease was bigger in the high rGTP group than in
the normal rGTP group. The results indicated that
there was large individual variability in the responsesof the risk factors for coronary heart disease to drinking. Subjects whose rGTP responds less to drinking
may have less disadvantageous effects of drinking.
2-1-2 Is there any good way of drinking for health?
We performed a cross-sectional study of 3660 maleworkers to examine whether the weekly frequency ofdrinking affects clinical laboratory data when theweekly amount of drinking is the same. The results
suggested that the weekly frequency of drinking
might affect the levels of HDL-cholesterol independently of the weekly alcohol consumption.
2-1-3 Are there the genetic factors responsible forindividual varieties regarding the response ofclinical laboratory data to alcohol consumption?
We examined 133 male workers who drank >300g of alcohol per week to sudy the relationshipbetween clinical laboratory data and the genetic polymorphisms of alcohol dehydrogenase (ADH) 2 andaldehyde dehydrogenase (ALDH) 2. The results suggested that the ADH2 genotype influences the
responses of blood pressure, triglycerides, and uricacid to alcohol consumption, while the ALDH2 geno-

type influences the response of the erythrocyte meancell volume to alcohol consumption.
2-2 Others (HbA1C, proteinuria, leptin)
2-2-1 HbA1C
HbA1C has been suggested to deteriorate with age.
However, controversy exists as to whether this deterioration is a consequence of the aging process itself orof variables related to aging, such as concomitant disease, medication, body-mass index, physical activity,
and the change in distribution of body fat. We performed a cross-sectional survey of 7664 male Japanese
workers aged 20-59 years. This survey suggested that
HbA1C increased with age itself, and body-massindex and a hereditary predisposition to diabetes
affected this age-dependent increase in HbA1C.
2-2-2 proteinuria
We performed a cross-sectional survey of 5174
male Japanese workers. Blood pressure and a family
history of diabetes were independent factors associated with proteinuria in subjects with a HbA1C below
5.9% who were not under medication for diabetes. In
contrast, HbA1C, obesity and smoking were associated with proteinuria in subjects who were under medication for diabetes and/or have a HbA1C above 5.9%.
These findings suggest that maintaining a HbA1C
level below 5.9%, non-smoking and a standard bodyweight may reduce the prevalence of proteinuria inJapanese men. Healthy life–style habits and standard
body weight are especially important for subjects
with a family history of diabetes.
We also performed a cross-sectional study of 237
men to examine the relationship between proteinuria
determined with a reagent strip and the angiotensin
converting enzyme (ACE) genotypes. The results suggest that the ACE DD genotype may be an independent risk factor of proteinuria in Japanese men.
2-2-3 lepein
Leptin, the ob gene product secreted by adipocyte,
decreases food intake while it increases energy expenditure and functions as an important signal for the
regulation of body weight in mouse. To investigate
whether smoking affects the serum level of leptin, we
performed a cross-sectional study of 708 male workers. The findings suggested that among life-style
habits, physical activity, but not smoking or alcohol
consumption, significantly affects the serum level of
leptin in Japanese men.
The leptin receptor is an isoform of the B219 geneproduct, a member of the hematopoietin receptor
family, which is expressed in very primitive
hematopoietic cells. Recent studies showed that leptin
plus erythropoietin acted synergistically to increase

189

erythroid development in vitro. These finding led us
to examine the relationship between the serum levelsof leptin and hemoglobin. The results of the above
cross-sectional study showed that the serum levels of
leptin is negatively associated with those of hemoglobin. Although the effect of leptin on hematopoiesis
may be modest, the results of our epidemiologic
study, together with the previous studies performedin vitro, suggest that leptin may play some role inhematopoiesis in humans.
3. Learning from patients at the points of biochemistry3-1 Study of apolipoprotein A-1 deficiency
We evaluated a 69-year-old Japanese woman withapolipoprotein (apo) A-I deficiency, high levels of
LDL-cholesterol, hypertension and impaired glucose
tolerance. The patient had corneal opacity, but neither
xanthomas, xanthelasma, nor tonsillar hypertrophy.
She was not symptomatic for coronary heart disease,and had normal electrocardiograms at rest and during
exercise on a cycle ergometer. She had severely
reduced levels of HDL-cholesterol (0.10-0.18 mmol/L)and no apo A-I (<0.6 mg/dL). LDL-cholesterol and apo
B as well as apo E were increased even under treatment with 10 mg pravastatin per day. Gel filtration
chromatography revealed that in addition to VLDL
and LDL fractions, she had apo A-II rich and apo E
rich fractions, which were present in the HDL fraction
separated by ultracentrifugation. A cytosine deletion
was identified by genomic DNA sequencing of the apo
A-I gene of the patient at the third base of codon 184
in the fourth exon, which led to a frame shift mutation and early termination at codon 200. This patientis the oldest among those with apo A-I deficiency
reported in the literature, and she had no symptomsof coronary heart disease despite the accumulated risk
for the disease.
3-2 Study of phytosterolemia
We diagnosed two women with multiple xanthomas, intermittent arthritis and thrombocytopenia
as phytosterolemia, an autosomal-recessive lipid storage disease, based on their increased serum concentrations of β-sitosterl, campesterol and sitostanol.
The gene responsible for this disease is located within
a distance of 18 cM between microssatellite markers
of D2S1788 and D2S1352 at chromosome 2p21. We
genotyped the patients and their family memberswith 16 microsatellite markers around this locus. The
results from the homozygosity mapping of one family
suggested that the gene was located within the distance of 12.6 cM between D2S2328 and D2S1352. We
have shortened the genetic distance by 5.4 cM.

190

3-3 Another good effect of pioglitazone, aninsulin sensitizing drug
The findings obtained from two patients with diabetes and asthma suggested that pioglitazone may
ameliorate symptoms of asthma. Pioglitazone is oneof the thiazolidinedione compounds which have beenused as antidiabetic drugs. Recent studies haverevealed that thiazolidinedione compounds also havevarious nonhypoglycemic effects, such as antiinflamatory, antiatherosclerotic and anticancer effects, in cultured cells and in experimental animal models.
Elucidation of the mechanism may enable the designof a novel class of drugs for asthma.
4. Researches in physiological fields
4-1 Cardiac functions using ultrasoundMajor research interests are evaluation of left ventricular and left atrial functions, detection of tissue
ischemia using contrast echo, and assessment of cardiac state during inactivity.
4-2 Influence of diesel exhaust particles on respiratory functions
We reported that diesel exhaust particles (DEP)
induced nuclear factor-kappa B(NFκB) activation,
upregulated expression of intercellular adhesion molecule(ICAM)-1 in human bronchial epithelial cells, andbenzene-extracted components are important for the
major activity of DEP. Increased expression of transforming growth factor-β1 in small airway epithelium
was also demonstrated.
4-3 Investigations on brain functions using magnetoencephalography (MEG)
We installed the MEG system, which is a newly
developed tool for neurophysiology, and studied
somatosensory evoked magnetic fields to on- and offsets of rectangular pressure stimuli.

Current Research
1. Quantification of cell surface antigens using
flow cytometry
Conventional methods of flow cytometry provide
us only a positive percentage of cells. If we can measure and obtain cell surface antigen density, moredetail analysis must be possible. The purpose of thisproject is quantification of cell surface antigens withflow cytometry.
2. Analysis of Th1/Th2, Tc1/Tc2 lymphocytes
using flow cytometry
A method to analyze blood samples with EDTA-2K

was developed. Infectious diseases and immunocompromised patients are under investigation.

damage, especially renal damage, we are now investigating new marker in urine from various patients.

3. Relationship between genetic polymorphismsof alcohol metabolizing enzymes, and diabetic
control and the risk factors for CHD in patientswith diabetes.
Subjects: Non-drinkers and drinkers with diabetes
Methods: Genetic polymorphisms of ADH2 and
ALDH2 are examined by the PCR.
Objective:
1) Is the effect of alcohol drinking on diabetic control
associated with the genetic polymorphisms of
ADH2 and ALDH2?
2) Is the effect of alcohol drinking on the risk factors
for CHD associated with the genetic polymorphisms of ADH2 and ALDH2?
3) Are those effects observed only in drinkers andnot in non-drinkers?

7. Analysis of perfusion and function of the left
ventricule
Myocardial perfusion is assessed by myocardial
contrast echo while regional left ventricular function
is evaluated by strain data derived from myocardial
Doppler velocity information.

4. Identification of the gene mutation responsible
for phytosterolemia in our three patients.
Recent studies showed that phytosterolemia might
be explained by mutations of the genes that encode
new members of the ATP-binding cassette (ABC)transporter family, ABCG5 and ABCG8. Now we are
examining whether our patients have the gene mutations of ABCG5 and ABCG8.
5. Physiological roles of FGF-23
A new comer to our Department has cloned andidentified FGF-23 as a causative factors for tumorinduced hypophosphatemic rickets/osteomalacia. We
have also established an ELISA assay for biologically
active full-length FGF-23. We are now investigating
physiological roles of FGF-23 in phosphate and vitamin D metabolism.
6. Adrenomodullin
As another new comer participated in our
Department, this new project is started. He has been
studying the physiological role of vasoactive peptide
to regulate blood pressure by modulating sympathetic
tone and its possible role in preventing organ damage.
Adrenomedullin, a vasoactive peptide was discoveredfrom pheochromocytoma cells and it is known toexist not only adrenal cells but also vascular smoothmuscle cells, endothelial cells, and other organs. He
and his colleagues have generated adrenomedullinknockout mice and showed that adrenomedullin prevents organ damage by reducing oxidative stress. In
clinical study, they have shown adrenomedullin isbetter indicator of circulating blood volume than ANP
in patients under hemodialysis. The measurement ofblood adrenomedullin level will be clinically important marker. On the other hand, to diagnose organ

8. Relationship between respiratory function andvarious pathophysiological condition
The influence of sleep apnea syndrome, lung cancer, asthma, aging or respiratory rehabilitation on pulmonary function has been investigated by a memberof our Department.
9. Study using magnetoencephalography (MEG)
We have not only been making efforts to obtainreimbursement for clinical MEG studies but also beenorganizing several research groups which consists ofmedical doctors from the department of neurology,
physiology, neurosurgery, pediatrics and otolaryngology in the University Hospital and researchers from
University of Tokyo School of Medicine. Current
themes are follows.
1) Epilepsy evaluation-spike source localization
2) Pre-surgical functional mapping-somatosensory,
motor, auditory, visual and language
3) Auditory evoked magnetic response as a measureof sensory gating in schizophrenia
4) Abnormality of mismatch negativity in schizophrenic patients
PromoteFuture Prospects
We have a future planning in each research theme
as follows.
1) Apply the quantity analysis method of cell surface
antigens with flow cytometry to actual clinicalsamples and evaluate its usefulness.
2) Monitor immunological status of immunocompromised patients and patients with infectious diseases by using lymphocyte activation markers.
3) Prospective study on the relation between the mortality due to CHD and alcohol drinking, whichtakes genetic polymorphisms of alcohol metabolizing enzymes into consideration4) Elucidation of the structures and functions of
ABCG5 and ABCG8 and design of a novel class ofdrugs for hypercholesterolemia5) Study on physiological and pathophysiologocalroles of FGF-23

191

6) Find new markers to diagnose early phase of renal
damage in diabetes or hypertensive patients aswell as to diagnose response to treatment.
7) Three dimensional analysis of left ventricular perfusion and function using cardiac ultrasonography8) Promote EEG and MEG studies of higher cognitive
brain function, especially focused on the investigation of gamma-band activities which are thought
to play a key roll in perception and cognition.
It might be said that the research themes of ourdepartment are diverse and contain a wide range ofsubjects. Although we have actually a lot of research
projects simultaneously, we can say that it is the characteristic of our department and our common themeis laboratory examination and the key word is laboratory medicine. The purpose of our research is to apply
our research products to actual clinical fields, and contribute to human health and welfare. But today’s
problem in our department is the shortage of thenumber of graduate students. We must make an effort
to obtain them.

Research Grants
1. Grant-in-Aid for Scientific Research (B) from The
Ministry of Education, Science, Sports and Culture(1998~2000) : The new application of flow cytometry – Quantification of cell surface antigens and its
clinical application –2. Grant-in-Aid for Scientific Research (B) from The
Ministry of Education, Science, Sports and Culture(2001~2003) : Practical utilization of selective cell
information using flow cytometry and its clinicalapplication
3. Grant-in-Aid for Scientific Research (C) from The
Ministry of Education, Science, Sports andCulture(1999/2000) : Studies on phytosterolemia:
Identification of the pathologic gene and examination of its pathogenesis
4. Grant-in-Aid for Scientific Research (C) from The
Ministry of Education, Science, Sports and Culture(2001/2002) : Reverse cholesterol transport inapolipoprotein A1 deficiency
5. Grants-in-aid for Promotion of AIDS Research from
the Ministry of Health and Welfare of Japan(2000~2002)

Select Publications
1. Ｎakahara, K., Ohashi, T., Oda, T., Hirano, T., Kasai,
M., Okumura, Ｋ.＆ Tada, T. : Asialo GM１as a cellsurface marker detected in acute lymphoblastic
leukemia. New England J Medicine. 302：674-677,
1980.
2. Ｎakahara, Ｋ., Ohashi, T., Ichimaru M., Amagasaki,T., Shimoyama, M., Nakauchil ＆ kumura, K. :Analyses of adult T-cell leukemia using the mono-

192

clonal (anti-Leu-l, anti-Leu-2a and anti-Leu-3a) andheterologous anti-glycolipid (anti-asialo GML) antlbodies. Clinical Immunology and Imunopathology,
25：43-52, 1982.
3. Yoneyama, A., Nakahara, K., Higashihara, M. ＆Kurokawa : Increased levels of soluble CD8 and CD4
inpatients with infectious mononucleosis. Brit J
Haematol 89 : 47-54, 1995.
4. Aoki, K., Suzuki, K., Sugano, A., Tasaka, T., Nakahara,K., Kuge, O. Omori, A. ＆ Kasai, M. : A novel gene,Translin, encodes a recombination hotspott binding
protein associated with chromosomal translation.
Nature Genetics lO: 167-174, 1995.
5. Aoki, I., Homori, M., Nakahara, K., Higashi, K. ＆Ishikawa : Effects of recombinant humaninterleukin-1 (rhIL-), -1 (rhIL-1) and rhIL-1 receptor
antagonist (rhIL1ra) on erythroid progenitors(CFU-E
and BFU-E) in human bone marrow. Exp
Hematology 23(3):217-25, 1995.
6. Mashige, F., Matsushima, Y., Miyata, C., Yamada, R.,
Kanazawa, H. Sakuma, I., Takai, N., Shinozuka, N.,
Ohkubo, A and Nakahara, K. : Simultaneous determination of catecholamines, their basic metabolites
and serotonin in urine by high-perfomance liquid
chromatography using a mixed-mode column and aneight-channel electrochemical detector. Biomedical
Chromatography. 9 : 221-225, 1995.
7. Watanabe, T., Matsushima, S., Okazaki, M.,
Nagamatsu, S., Hirosawa, K., Uchimura, H. andNakahara, K. : Localization and ontogeny of GLUT3
expression in the rat retina. Developmental Brain
Research. 94, 60-66, 1996.
8. Watanabe, T., J. T. Voyvodic., T. Chan-Ling., Sagara,
H., Hirosawa, K., Mio, Y., Matsushima, S., Uchimura,H., Nakahara, K. and Martin Raff: Differentiationand morphogenesis in pellet culture of developing rat
retinal cells. The Jounal of Comparative Neurology.
377: 341-350, 1997.
9. Hashimoto Y, Togo M, Sato H, Hashimoto N,
Watanabe T, Kurokawa K, and Nakahara K.
Characteristics of protein kinase C-independent exocytosis in human platelets. Thromb Res 88: 51-58,
1997
10. Aoki, K. Inazawa, J., Takahashi, T., Nakahara, K.,
and Kasai, M. : Genomic structure and chromosomal
localization of the gene encoding translin, a recombination hotspot bindlng protein. Genomics, 43 (2) :237-241, 1997.
11. Hashimoto,Y., Togo,M., Sato,H., Hashimoto,N.,
Watanabe,T., Kurokawa,K. and Nakahara,K. :Characteristics of protein kinase C-dependent exocytosis in human platelets. Thrombosis Research, 88(1) : 51-58, 1997.
12. Sakurai, S., Shiojima, I., Tanigawa, T., and Nakahara,
K. : Aminoglycosides prevent and dissociate theaggregation of platelets in patients with EDTAdependent pseudothrombocytopenia. British J. of
Haematology,99: 817-823,1997.
13. Koike, Y., Yoneyama, A., Shirai, J., Ishida, T., Shoda,E., Miyazaki, K., Sunaga, S., Horie, R., Aoki, K.,Koike, K., Ogata, I., Takahara, T., Kato, T., Nakahara,K., Kariya, T., and Higashihara, M. : Evaluation ofthrombopoiesis in thrombocytopenic disorders by
simultaneous measurement of reticulated platelets ofwhole blood and serum thrombopoietin concentrations. Thromb Haemost. 79:1106-1110,1998.
14. Kawasaki, S., Takizawa, H., Ohtoshi, T., Takeuchi,N., Kohyama, T., Nakamura, H., Asama, T.,
Kobayashi,K., Nakahara, K., Morita, Y. And
Yamamoto, K. : Roxithromycin in hibits cytokine
production by and neutrophil attachment to humanbronchial epithelial cells in vitro. American Agentsand Chemotherapy. 42 (6) 1499-1502, 1998.
15. Yan, K., Nakahara, K., Awa, S., Nishibori, Y.,
Nakajima, N., Kataoka, S. Maeda, M., Watanabe, T.,
Matsushima, S., and Watanabe, N. : The increase ofmemory T cell subsets in children with idiopathic
nephrotic syndrome. Nephron.79, 274-27８, 1998.
16. Aoki, K., Meng, G., Suzuki, K., Takashi, T., Kameoka,
Y., Nakahara, K., Ishida, R., Kasai, M. : RP58 associates with condensed chromatin and mediates asequence-specific ranscriptional repression. The
Journal of Biological Chemistry. 273 (41), Issue ofOctober 9, 26698-26704, 1998.
17. Hashimoto, Y., Hutamura, A., Watanabe, N., Togo,
M., Sato, H., Hara, M., Tsukamoto, K., Kimura, S.,
and Nakahara, K. : Relationship between glcosylated
hemoglobin and the prevalence of proteinuria inJapanese men. Internal Medicine, 38 (1), 6-11,1999.
18. Hashimoto, Y., Futamura, A., Watanabe, N., Togo,
M., Sato, H., Hara, M., Tsukamoto, K., imura, S., andNakahara, K. : Relationship between glycosylated
hemoglobin and the prevalence of roteinuria inJapanese men. Intern Med., 38(1):6-11, 1999.
19.Togo, M., Tsukamoto, K., Satoh, H., Hara, M.,
Futamura, A., Nakarai, H, Nakahara, K., Hashimoto,Y. : Relationship Between Levels of Leptin and
Hemoglobin in Japanese Men. Blood., 93:4444-4445,
1999.
20. Okubo, S., Mashige, F., Hashimoto, Y., Nakahara, K.,
Kanazawa, H., Matsushima, Y., Fukushima, T.,
Huang, Y., and Imai,I.;D-Serine concentration inblood serum of liver transplanted patient.
Chromatography, 21:43-47,2000.
21. Takizawa, H., Abe, S., Ohtoshi, T., Kawasaki, S.,
Takami, K., Desaki, M., Sugawara, I., Hashimoto, S.,
Azuma, A., Nakahara, K. and Kudoh, S. : Diesel
exhaust particles up-regulate expression of intercellular adhesion molecule-1 (ICAM-1) in human epithelial cells. Clin Exp Immunol 120: 356-362, 2000.
22. Meng, G., Inazawa, J., Ishida, R., Tokura, K.,
Nakahara, K., Aoki, K. & Kasai, M. : Structural analysis of the gene encoding RP58, a sequence-specifictransrepressor associated with heterochromatin.
Gene 242, 59-64, 2000.
23. Meng, G., Aoki, K., Tokura, K., Nakahara, K.,
Inazawa, Johji. & Kasai, M. : Genomic structure andchromosomal location of the gene encoding TRAX, a
Translin-associated factor X. J Hum Genet 45: 305308, 2000.

24. Okubo, S., Mashige, F., Omori, M., Hashimoto, Y.,
Nakahara, K., Kanazawa, H. & Matsushima, Y. :Enantiomeric determination of L-and D-lactic acid inhuman cerebrospinal fluid by chiral ligand exchange
high-performance liquid chromatography. Biomed
Chromotogr 14: 474-477, 2000.
25. Watanabe, T. and Nakahara, K. : Quantification ofcell surface molecule density by flow cytometry.
Symposium Abstract, The Sixth Asian Conference ofClinical Pathology. S89, 2000.
26. Yumoto, M., Saitoh, O., Kaneko, Y., Anami, K.,
Matsuda, H., Ugawa, Y. and Nakahara, K. :Somotosensory evoked magnetic fields to on-and offsets of rectangular pressure stimuli. Proc. of 12th
International Conference on Biomagnetism,243-246,
2000.
27. Togo, M., Hashimoto, Y., Futamura, A., Tsukamoto,K., Satoh, H., Watanabe, T., Nakarai, H., Nakahara,
K.,and Kimura, S. : Relationship between serum levelof leptin and life-style habits in Japanese men. Horm
Res 54,169-173,2000.
28. Togo, M., Toda, T., L.A.Nguyen, Kubota, S.,
Tsukamoto, K., Satoh, H., Hara, M., Iso-o,N., Noto,
H., Kimura, S., Nakahara, K., Seyama, Y. andHashimoto, Y. : Genetic analysis of phytosterolaemia. J. Inherit. Metab. Dis. 24, 43-50, 2001.
29. Sakurai,S., Takenaka,K., Shiojima, I., Watanabe,F.,
Sonoda, M., Uno,K., Kuwada,Y., Nakahara,K., andNagai,R.: Traumatic Tricuspid Insufficiency.
Echocardiography:A J. of CV Ultrasound & Allied
Tech. 18 (4) 303-304, 2001.
30. Kawasaki, S., Takizawa, H., Takami, K., Desaki, M.,
Okazaki, H., Kasama, T., Kobayashi, K., KYamamoto,K., Nakahara, K., Tanaka, M., Sagai, M., andOhtoshi, T. : Benzene-extracted components areimportant for the major activity of diesel exhaust
particles (DEP): Effect on IL-8 gene expression inhuman bronchial epithelial cells. Am J Repir Cell Mol
Biol 24, 419-426, 2001.
31. Hashimoto, Y., Futamura, A., Nakarai, H., Yokota,
H., Omura, M., Tsukamoto, K., Togo, M., Sato, H.,
Hara, M., Isoo, N., Taguchi, J., and Nakahara, K. :Association between deletion polymorphism ofangiotensin converting enzyme gene and proteinuria
in Japanese overweight men., J Occup Health.43,8084, 2001.
32. Sakurai, S., Takenaka, K., Nakahara, K., and Nagai,
R. : Traumatic tricuspid insufficiency.
Echocardiography 18 : 303, 2001.
33. Takizawa, H., Tanaka, M., Takami, K., Ohtoshi, T.,
Ito, K., Satoh, M., Okada, Y., Yamasawa, F., Umeda,
A., and Nakahara, K. : Increased Expression ofTransforming Growth Factor-β1 in Small Airway
Epithelium from Tobacco Smokers and Patients with
Chronic Obstructive Pulmonary Disease (COPD). Am
J Respir Crit Care Med 163, 1476-1483, 2001.
34. Mashige. F., Tsuno H. N., Maekawa, M., Okada, S.,
Tanaka, N., Matsui, S., Watanabe, S. and Nakahara,
K. : Noninvasive combination assay of free L-fucose
and suflated bile acids in urine for diagnosis of liver193

and pancreatic diseases and solid tumor. Journal of
Analytical Bio-Science 24(4) 317-323, 2001.
35. Hashimoto,Y., Futamura, A., Nakarai, H., andNakahara, K. : Relationship between response of glutamly transpetidase to alcohol drinking and risk
factors for coronary heart disease. Athreosclerosis
158 465-470, 2001.
36. Sakurai, S., Takenaka, K., Shiojima I., Watanabe, F.,
Onoda, M., Uno, K., Kuwada,Y., Nakahara, K., andNagai, R. : Traumatic Tricuspid Insufficiency.
Echocardiography:A J. of CV Ultrasound & Allied
Tech. 18 (4) 303-304, 2001.
37. Kawasaki, S., Takizawa, H., Takami, K., Desaki, M.,
Okazaki, H., Kasama, T., Kobayashi, K., Yamamoto,K., Nakahara, K., Tanaka, M., Sagai, M., andOhtoshi, T. : Benzene-extracted components areimportant for the major activity of diesel exhaust
particles : Effect on interleukin-8 gene expression inhuman bronchial epithelial cells. Am J Respir Cell
Mol Biol. 24, 419-426, 2001.
38. Hashimoto, Y., Futamura, A., Nakarai, H., andNakahara, K. : Effects of the frequency of alcohol
intake on risk factors for coronary heart disease.
European Journal of Epidemiology 17, 307-312, 2001.
39. Sakatani, T., Shimazaki, C., Hirai, H., Okano, A.,
Hatsuse, M., Okamoto, A., Takahashi, R., Ashihara,E., Inaba, T., Yokota, H., Nakahara, K., Hirai, H. andNakagawa, M. : Early relapse after hgh-dose
chemotherapy rescued by tumor-free autologous
peripheral blood stem cells in acute lymphoblastic
leukemia : Importance of monitoring for WT1mRNA quantitatively. Leukemia and Lymphoma42(1-2), 225-229, 2001.
40. Yokota, H., Tsuno, N. H., Tanaka, Y., Fukui, T.,
Kitamura, K., Hirai, H., Osumi, K., Itou, N., H. Satoh,
M. Okabe, S, Nakahara, K. : Quantification of
Minimal Residual Disease in Patients with ela2 BCRABL Positive Acute Lymphoblastic Leukemia Using a
Real-Time RT-PCR Assay. Leukemia(16), 1167-1175,
2002.
41. Yokota H, Hashimoto Y, Okubo S, Yumoto M,
Mashige F, Kawamura M, Kotani K, Usuki Y,
Shimada S, Kitamura K, Nakahara, K. Apolipoprotein
A-I deficiency with accumulated risk for CHD but nosymptoms of CHD. Atherosclerosis 162: 399-407,
2002
42. Hashimoto, Y., and Nakahara, K. : Improve of asthma after administration of pioglitazone. Diabetes
Care, 25(2),401, 2002.
43. Kanazawa, H., Okada, A., Matsushima, Y., Yokota,
H., Okubo, S., Mashige, F., and Nakahara, K. :Determination of omeprazole and its metabolites inhuman plasma by liquid chromatograpy-mass spectrometry. Journal of Chromatgrahy A, 949, 1-9, 2002.
44. Ono, Y., Mashige, F., Sano, S., Nanba, Y., Okubo, S.,
Hashimoto, Y., and Nakahara, K. : α 1Antichymotrypsin in serum and cerebrospinal fluid
Part1 : Development of latex agglutination test for
determination of α 1-antichymotrypsin. Journal of
Analytical Bio-Science, 25(2), 153-158, 2002.

194

45. Mashige, F., Ono, Y., Nanba, Y., Okubo, S.,
Hashimoto Y., and Nakahara, K. : α 1Antichymotrypsin in serum and cerebrospinal fluid
Part2 : Clinical implications of α 1-antichymotrypsin
and other acute phase reactant proteins in
Alzheimer-type dementia and other diseases. Journalof Analytical Bio-Science, 25(2), 159-163, 2002.
46. Kitamura, K., Miwa, S., Ikemoto, T., Tatsumi, N.,
Nakahara, K., Higashi, K., and Watanabe, K. :Proposal of guidelines for surface antigen analysis ofperipheral lymphocytes using flow cytometry andpreliminary quality control. Global Standardization
and Advanced quality Management’01, Eibun Press,
Ltd., Osaka, Japan, pp48, 2002.
47. Hashimoto,Y., Nakayama, T., Futamura, A., Omura
M., and Nakahara, K. : Erythrocytes mean cell volume and genetic polymoprphism of aldehyde dehydrogenase 2 in alcohol drinkers. Blood 99(9):3487-8,
2002.
48. Hashimoto, Y., Nakayama, T., Futamura, A., Omura,M., Nakarai, H., and Nakahara, K. : Relationshipbetween genetic polymorphisms of alcohol-metabolizing enzymes and changes in risk factors for coronary heart disease associated with alcohol consumption. Clinical Chemistry, 48(7), 1043-1048, 2002.
49. Takami, K., Takuwa, N., Okazaki, H., Kobayashi, M.,
Ohtoshi, T., Kawasaki, S., Dohi, M., Yamamoto, K.,
Nakamura, T., Tanaka, M., Nakahara, K., Takuwa,
Y., and Takizawa, H. : Interferon-gamma Inhibits
hepatocyte growth factor-stimulated cell proliferation of human bronchial epithelial cells upregulationof p27kip1 cyclin-dependent kinase inhibitor.
American Journal of Respiratory Cell & Molecular
Biology, 26(2), 231-238, 2002
50. Takenaka, K., Suzuki, Y., Uno, K., Sato, M., Komuro,T., Haruna Y., Kobayashi, H., Kawakubo, K., Sonoda,
M., Asakawa, M., Nakahara, K., and Gunji, A. :Effects of rapid saline infusion on orthostatic intolrerance and automatic tone after 20 days bed rest.
Am J Cardiol 89, 557-561, 2002.

Department ofTransfusion Medicine
Outline and Research Objectives
The Department of Transfusion Medicine was established in 1949, and actually is composed by 4 medical doctors, 10 medical technicians, 1 nurse and 1 office assistant.

Faculties and Students
Professor and Chair

Yoichi Shibata, M.D., Ph.D. (sinceApril 1993)
Associate Professors absent
Lecturer
absentAssociate .................................3
Research Students...................4 research students fromthe department ofSurgical Oncology aredeveloping their research
projects at the department’s laboratory.
Secretary .................................1
Medical Technician...............10
Nurse.......................................1

4)

5)

6)

Past Research and Major Accomplishments
The major research projects developed at the
department are related to:1) Human platelet antigen/antibody detection andcharacterization: the mixed-passive hemagglutination (MPHA) assay, a serological method largely
used in Japan for the screening and characterization of platelet alloantigens and alloantibodies,
was developed at the department by Yoichi
Shibata. Using this method, a new platelet
alloantigen system, namely the HPA-4 system(Yuka/b), was discovered by Y. Shibata, et al.
2) Human endothelial cell antigen/antibody detection
and characterization: the MPHA method, developed by Y. Shibata, et al., was modified for thedetection and characterization of endothelial cellalloantigens and alloantibodies. Using thismethod, monoclonal antibodies reactive withtumor endothelial cells were developed at the
department. This monoclonals are being investigated for a possible application in cancer treatment.
3) Immunotherapy of cancer using LAK cells anddendritic cells: the effectiveness of lymphokineactivated killer (LAK) cells, as well as of peripheral
blood and monocyte-derived dendritic cells for
immunotherapy of cancer was investigated in

7)

8)

vitro, and were tested in a clinical trial for thetreatment of patients with gastrointestinal cancer
and brain tumors.
Development of new vaccines targeting the tumor
angiogenesis for the treatment of cancer: new vaccine strategies for the treatment of solid tumors
by targeting the tumor vasculature are being
developed at the department. The vaccine, tested
in an animal model of colon cancer metastasis,
showed to be very effective, and is being prepared
for the clinical trial.
Vaccination therapy of patients with habitualabortion using the husband’s lymphocytes, andinvestigation on their mechanism of action.
Effects of radiation and filtration on platelet function: the effects of radiation and the use of whiteblood cell (WBC)-reduction filters on the platelet
function (adhesion and aggregation) have beenstudied. Irradiation did not affect the platelet
function, but the WBC-reduction filters, depending on the type of filter, reduced the adhesive
capacity of platelets.
Development of a new methodology for the evaluation of platelet function: a new method, based on
the adhesive capacity of platelets, is being developed at the department.
Peripheral blood stem cells transplantation:
peripheral blood stem cells are isolated at the
department for transplantation in leukemic
patients.

Current Research
The research projects described above are stillongoing at the department of Transfusion Medicine,which are:1) Human platelet antigen/antibody detection andcharacterization, and study on their role in transfusion practice;2) Human endothelial cell antigen/antibody detection
and characterization, and study on their role in
the pathogenesis of inflammatory and autoimmune diseases, as well as in organ transplantation;3) Immunotherapy of cancer using LAK cells and dendritic cells;

1954) Development of new vaccines targeting the tumor
angiogenesis for the treatment of cancer;
5) Lymphocyte vaccination therapy for patients with
habitual abortion;
6) Effects of radiation and filtration on platelet function;7) Development of a new methodology for the evaluation of platelet function;
Peripheral blood stem cells transplantation.

Future Prospects
The future prospects of the department are as follows:
1) To develop the ideal method of platelet collection
and preservation for transfusion, especially related
to preservation of platelets without loss of theirfunction;
2) To develop a good methodology for the evaluationof platelet function;3) To develop a new therapeutic strategy to combat
cancer by targeting the angiogenic vasculature oftumors;4) To develop the vascular regeneration medicine
using peripheral blood and cord blood stem cells.

Research Grants(1) Grant-in-Aid for Scientific Research from the Ministryof Education, Culture, Sports, Science andTechnology of Japan –Kiban (C)(2); Development of a
new methodology for the detection of anti-endothelial cell antibodies and evaluation on their clinicalsignificance. 1997 – 1998;3 million yen.
(2) Grant-in-Aid for Scientific Research from the Ministryof Education, Culture, Sports, Science andTechnology of Japan (B)(2): Development of a monoclonal antibody-based strategy for cancer treatment
targeting the angiogenic vascular endothelium. 1999
– 2001;11,800 thousand yen.
(3) Grant-in-Aid for Scientific Research from the Ministryof Education, Culture, Sports, Science andTechnology of Japan- Grant for Highly-advanced
Medicine, Development of a cell-based and genebased strategies for the treatment of malignant andother life-threatening disorder of difficult control.
1998 – 2000; 131,304 thousand yen.

Select Publications
1. Osada T, Fujimaki T, Takamizawa M, Tsuno NH,
Kirino T, Shibata Y. Dendritic cells activate antitumor immunity for malignant intracranial germ celltumor: a case report. Japanese Journal of ClinicalOncology;31(8):403-6, 2001.
2. Osada T, Nagawa H, Kitayama J, Tsuno NH, Ishihara
S, Takamizawa M, Shibata Y. Peripheral blood dendritic cells, but not monocyte-derived dendritic cells,

196

can augment human NK cell function. Cell Immunol,;213(1):14-23, 2001.
3. Maejima M, Fujii T, Yamashita T, Hara N, Hamai Y,
Miki A, Kozuma S, Okai T, Shibata Y, Taketani Y.
Immunotherapy before and during pregnancy
improvespregnancy outcome in women who sufferfrom recurrent abortion and did not benefit from
immunotherapy before pregnancy. American Journalof Reproductive Immunology;39(1):12-5, 1998.
4. Maejima M, Fujii T, Okai T, Kozuma S, Shibata Y,
Taketani Y. Beta2-glycoprotein I-dependent anticardiolipin antibody in early recurrent spontaneousabortion. Human Reproduction;12(10):2140-2, 1997.
5. Maejima M, Fujii T, Kozuma S, Okai T, Shibata Y,
Taketani Y. Presence of HLA-G-expressing cells modulates the ability of peripheral blood mononuclear
cells to release cytokines. American Journal of
Reproductive Immunology;38(2):79-82, 1997.
6. Minchinton RM, Dawkins B, Chynoweth L, Pearson
H, Lown JA, Shibata Y. In pursuit of enigmatic
platelet antibodies--anti-HPA-2b and anti-HPA-3a.
Transfusion Medicine;6(3):289-91, 1996.
7. Takeuchi F, Kuwata S, Nakano K, Nabeta H, Hong
GH, Shibata Y, Tanimoto K, Ito K. Association of
TAP1 and TAP2 with systemic sclerosis in Japanese.
Clinical & Experimental Rheumatology;14(5):513-21,
1996.
8. Fujii T, Tsushima R, Okai T, Shibata Y, Taketani Y.
Allo-non-specific elevation of maternal killer cell
activity in intrauterine growth restriction.
International Journal of Gynaecology & Obstetrics;
52(3):237-42, 1996.
9. Chiu L, Nishimura M, Ishii Y, Nieda M, Maeshima
M, Takedani Y, Shibata Y, Tadokoro K, Juji T.
Enhancement of the expression of progesterone
receptor on progesterone-treated lymphocytes after
immunotherapy in unexplained recurrent spontaneous abortion. American Journal of Reproductive
Immunology;35(6):552-7, 1996.
10. Sunami E, Tsuno NH, Kitayama J, Osada T, Saito S,
Tomozawa S, Tsuruo T, Shibata Y, Nagawa H.
Decreased Synthesis of MMP-7 and Adhesion to
Extracellular Matrix Proteins of Human Colon Cancer
Cells Treated with Troglitazone. Surgery Today 2001
11. Tomozawa S, Tsuno NH, Sunami E, Hatano K,
Kitayama J, Osada T, Saito S, Tsuruo T, Shibata Y,
Nagawa H. Cyclooxygenase-2 overexpression correlates with tumour recurrence, especially haematogenous metastasis, of colorectal cancer. Br J Cancer;
83(3):324-328, 2000.
12. Sunami E, Tsuno N, Osada T, Saito S, Kitayama J,
Tomozawa S, Tsuruo T, Shibata Y, Muto T, Nagawa
H. MMP-1 is a prognostic marker for hematogenous
metastasis of colorectal cancer. Oncologist;5(2):10814, 2000.
13. Saito S, Tsuno N, Nagawa H, Sunami E, Zhengxi J,
Osada T, Kitayama J, Shibata Y, Tsuruo T, Muto T.
Expression of platelet-derived endothelial cell
growth factor correlates with good prognosis inpatients with colorectal carcinoma. Cancer Jan
1;88(1):42-9, 2000.
14. Tomozawa S, Nagawa H, Tsuno N, Hatano K, OsadaT, Kitayama J, Sunami E, Nita ME, Ishihara S, Yano
H, Tsuruo T, Shibata Y, Muto T. Inhibition of
haematogenous metastasis of colon cancer in miceby a selective COX-2 inhibitor, JTE-522. British
Journal of Cancer; 81(8):1274-9, 1999.

197

Department of
Obstetrics and Gynecology
Outline and Research Objectives
The goal we aim at is the improvement of holistic women’s health. To achieve it, we
integrate all the knowledge in the field of reproductive medicine, perinatal medicine andgynecological oncology, which has been established on the basis of our history more thanhundred years, and will develop them.

Faculties and Students
Professor and Chair
Associate Professor
Lecturer

Yuji Taketani, MD, PhD (from 1994)
Shiro Kozuma
Toshiyuki Kojima
Koji Kugu
Associate ...............................15
Postdoctoral Fellow ................2
Graduate Student....................2
Research Student ..................15
Secretary .................................2

Past Research and Major Accomplishments
Professor Taketani has led the clinical and basic
research on reproductive endocrinology in Japan.
Especially, he is the distinguished leader in the
research field of endometriosis, and has achieved
many valuable studies. He has also conducted theresearch which went into the broad range in obstetrics and gynecology by his prominent leadership inthe department. There are more than 250 papers
which he directed in these two decades.

Current Research
We have a highly organized infertility clinic, where
every patient is systemically examined and after diagnosis of underlying infertility factor(s) appropriate
treatment is performed following our protocol. Onceit turns out higher level of treatment is necessary,
ART is applied to such cases. We have been engaged
in in vitro fertilization and embryo transfer (IVF-ET)
as a main axis of ART for fifteen years. Conventional
IVF-ET is mainly indicated to cases with tubal factor,
mild male factor, immunological factor or of unexplained infertility factor. In case of severe male factor
or other fertilization disorder intracytoplasmic sperm
injection (ICSI) is performed. Now we have about 300
cases of IVF-ET every year, which conventional IVFET and ICSI share almost equally. The pregnancy rateof conventional IVF-ET is around 25% per embryo

198transfer cycle, which is comparable with that of ICSI.
Other ART techniques such as embryo cryopreservation and assisted zona hatching are also under way.
Total number of delivery cases is 426 per year.
Recently cases of obstetrical emergencies like abruptio
placentae, eclampsia and uterine ruptures transported
from neighboring hospitals have been increasing. Ourservice is an important part of Tokyo Metropolitan
Service System for Maternal Welfare and Perinatal
Medicine.
Primary care for peri/post-menopausal women is
becoming more and more important. We have alreadyestablished the primary care system for women focusing on climacteric syndrome and osteoporosis.
Hormone replacement therapy (HRT) is employed forthe purpose.
In basic research section, a couple of projects asfollows are under way, some of which have alreadyyielded interesting findings; 1) the mechanism of folliculogenesis and follicular apoptosis in the ovary, 2)
the functions of gynecologic hormones such as
gonadotropins and ovarian steroids, 3) the biochemical evaluation of early embryos and establishment of anew procedure of embryo cryopreservation, 4) thegenetic mechanism in gamete formation and development and 5) effect of sex steroid on bone metabolism.

Future Prospects
To improve the pregnancy rate in the infertility
clinic, the basic research on both the embryo and theendometrium, ovarian function and its regulatory
mechanism must be proceed. To take the precautionsin the infertility treatment, the development of thesafe protocol and secure system is also demanded.
The validity of hormone replacement therapy for
managing peri/post-menopausal women is becoming a
subject of discussion. To seek the answer this problem, we are studying on the various protocols in clinical and by experiments.
In the obstetrical field, the researches on the
immunological feto-maternal interaction are going on.

In vivo study of fetal physiology using sheep model
are in progress, expected to provide valuable knowledge for clinical field.

Research Grants
Grant-in-Aid for Scientific Research, Japan Society
for the Promotion of Science: Scientific Research (A)“Mechanism of conception in terms of embryo-maternal interaction” (1999-2001)
Grant-in-Aid for Scientific Research, Japan Society
for the Promotion of Science: Scientific Research (B)“Risk factors associated with progression and recurrence of endometriosis” (2000-2002)
Grant-in-Aid for Scientific Research, Ministry of
Health and Welfare: “Status of endometriosis in Japanin terms of reproductive health” (1998)
Grant-in-Aid for Scientific Research, Ministry of
Health, Labor and Welfare: “Strategy for reproductive
health-keeping of women with menstrual disorders”(1999-2000)
Grant-in-Aid for Scientific Research, Ministry of
Health, Labor and Welfare: “Status of reproductive
health of women with reproductive disorders in thesociety” (2001-2002)

Select Publications (most recent 50 papers)
1. Hiroi H, Momoeda M, Yamauchi N, Abe Y,
Yoshikawa H, Tsutsumi O and Taketani Y. An earlier
menopause as clinical manifestation of granulosa-celltumor: a case report. J Obstet Gynaecol Res 26, 9-12,
2000.
2. Hyodo H, Ishikawa Y, Tsuneyama H, Kashiwase K,
Toyoda C, Uchikawa M, Akaza T, Fujii T, Kozuma S,
Taketani Y and Juji T. New RhD(IVb) identified inJapanese. Vox Sang 79, 116-7, 2000.
3. Hyodo H, Ishikawa Y, Kashiwase K, Ogawa A,
Watanabe Y, Tsuneyama H, Toyoda C, Uchikawa M,
Akaza T, Fujii T, Kozuma S, Taketani Y and Juji T.
Polymorphisms of RhD(Va) and a new RhD(Va)-likevariant found in Japanese individuals. Vox Sang 78,
122-5, 2000.
4. Kamei Y, Takeda Y, Teramoto K, Tsutsumi O,
Taketani Y and Watanabe K. Human NB-2 of thecontactin subgroup molecules: chromosomal localization of the gene (CNTN5) and distinct expression
pattern from other subgroup members. Genomics 69,
113-9, 2000.
5. Koga K, Osuga Y, Tsutsumi O, Okagaki R, Momoeda
M, Yano T, Fujiwara T, Takai Y, Kugu K, Morita Y
and Taketani Y. Increased concentrations of soluble
tumour necrosis factor receptor (sTNFR) I and II inperitoneal fluid from women with endometriosis.
Mol Hum Reprod 6, 929-33, 2000.
6. Koga K, Osuga Y, Tsutsumi O, Momoeda M, Suenaga
A, Kugu K, Fujiwara T, Takai Y, Yano T and Taketani
Y. Evidence for the presence of angiogenin in humanfollicular fluid and the up-regulation of its production
by human chorionic gonadotropin and hypoxia. J
Clin Endocrinol Metab 85, 3352-5, 2000.
7. Kuroda K, Kamei Y, Kozuma S, Kikuchi A, Fujii T,
Unno N, Baba K and Taketani Y. Cephalopagus conjoined twins. Ultrasound Obstet Gynecol 16, 293,
2000.
8. Kuroda K, Kamei Y, Kozuma S, Kikuchi A, Fujii T,
Unno N, Baba K and Taketani Y. Prenatal evaluationof cephalopagus conjoined twins by means of threedimensional ultrasound at 13 weeks of pregnancy.
Ultrasound Obstet Gynecol 16, 264-6, 2000.
9. Matsumi H, Yano T, Osuga Y, Kugu K, Tang X, Xu
JP, Yano N, Kurashima Y, Ogura T, Tsutsumi O, Koji
T, Esumi H and Taketani Y. Regulation of nitricoxide synthase to promote cytostasis in ovarian follicular development. Biol Reprod 63, 141-6, 2000.
10. Matsumoto K, Yoshikawa H, Nakagawa S, Tang X,
Yasugi T, Kawana K, Sekiya S, Hirai Y, Kukimoto I,
Kanda T and Taketani Y. Enhanced oncogenicity ofhuman papillomavirus type 16 (HPV16) variants inJapanese population. Cancer Lett 156, 159-65, 2000.
11. Matsumoto K, Kawana K, Yoshikawa H, Taketani Y,
Yoshiike K and Kanda T. DNA vaccination of micewith plasmid expressing human papillomavirus 6
major capsid protein L1 elicits type-specific antibodies neutralizing pseudovirions constructed in vitro. J
Med Virol 60, 200-4, 2000.
12. Miki A, Fujii T, Ishikawa Y, Hamai Y, Yamashita T,
Tadokoro K, Kozuma S, Juji T and Taketani Y.
Immunotherapy prevents recurrent abortion without
influencing natural killer receptor status. Am J
Reprod Immunol 43, 98-106, 2000.
13. Nakagawa S, Yoshikawa H, Yasugi T, Kimura M,
Kawana K, Matsumoto K, Yamada M, Onda T and
Taketani Y. Ubiquitous presence of E6 and E7 transcripts in human papillomavirus-positive cervical carcinomas regardless of its type. J Med Virol 62, 251-8,
2000.
14. Osuga Y, Koga K, Tsutsumi O, Igarashi T, Okagaki R,
Takai Y, Matsumi H, Hiroi H, Fujiwara T, Momoeda
M, Yano T and Taketani Y. Stem cell factor (SCF)
concentrations in peritoneal fluid of women with orwithout endometriosis. Am J Reprod Immunol 44,
231-5, 2000.
15. Wang Y, Yano T, Kikuchi A, Yano N, Matsumi H,
Ando K, Kasai Y, Watanabe M, Okagaki R, Osuga Y
and Taketani Y. Comparison of the effects of addback therapy with various natural oestrogens onbone metabolism in rats administered a long-acting
gonadotrophin-releasing hormone agonist. J
Endocrinol 165, 467-73, 2000.
16. Yano T, Radulovic S, Osuga Y, Kugu K, Yoshikawa H,
Taketani Y and Schally AV. Inhibition of humanepithelial ovarian cancer cell growth in vitro bysomatostatin analog RC-160. Oncology 59 Suppl 1,
45-9, 2000.
17. Yoshida S, Unno N, Kagawa H, Shinozuka N,
Kozuma S and Taketani Y. Prenatal detection of ahigh-risk group for intrauterine growth restriction

199

based on sonographic fetal biometry. Int J Gynaecol
Obstet 68, 225-32, 2000.
18. Yoshikawa H, Jimbo H, Okada S, Matsumoto K,
Onda T, Yasugi T and Taketani Y. Prevalence ofendometriosis in ovarian cancer. Gynecol Obstet
Invest 50 Suppl 1, 11-7, 2000.
19. Hiroi H, Kugu K, Hoshino H, Kozuma S and Taketani
Y. Hyperemesis gravidarum associated with thyrotoxicosis and a past history of an eating disorder.
Arch Gynecol Obstet 265, 228-30, 2001.
20. Hiroi H, Yasugi T, Matsumoto K, Fujii T, Watanabe
T, Yoshikawa H and Taketani Y. Mucinous adenocarcinoma arising in a neovagina using the sigmoid
colon thirty years after operation: a case report. J
Surg Oncol 77, 61-4, 2001.
21. Kanai T, Fujii T, Keicho N, Tokunaga K, Yamashita
T, Hyodo H, Miki A, Unno N, Kozuma S and
Taketani Y. Polymorphism of human leukocyte antigen-E gene in the Japanese population with or without recurrent abortion. Am J Reprod Immunol 45,
168-73, 2001.
22. Kanai T, Fujii T, Unno N, Yamashita T, Hyodo H,
Miki A, Hamai Y, Kozuma S and Taketani Y. Humanleukocyte antigen-G-expressing cells differently modulate the release of cytokines from mononuclear cells
present in the decidua versus peripheral blood. Am J
Reprod Immunol 45, 94-9, 2001.
23. Kanai T, Fujii T, Kozuma S, Yamashita T, Miki A,
Kikuchi A and Taketani Y. Soluble HLA-G influences
the release of cytokines from allogeneic peripheral
blood mononuclear cells in culture. Mol Hum Reprod
7, 195-200, 2001.
24. Kawana K, Kawana Y, Yoshikawa H, Taketani Y,
Yoshiike K and Kanda T. Nasal immunization ofmice with peptide having a cross-neutralization epitope on minor capsid protein L2 of human papillomavirus type 16 elicit systemic and mucosal antibodies. Vaccine 19, 1496-502, 2001.
25. Kawana Y, Kawana K, Yoshikawa H, Taketani Y,
Yoshiike K and Kanda T. Human papillomavirus type
16 minor capsid protein l2 N-terminal region containing a common neutralization epitope binds to thecell surface and enters the cytoplasm. J Virol 75,
2331-6, 2001.
26. Koga K, Osuga Y, Tsutsumi O, Yano T, Yoshino O,
Takai Y, Matsumi H, Hiroi H, Kugu K, Momoeda M,
Fujiwara T and Taketani Y. Demonstration of angiogenin in human endometrium and its enhanced
expression in endometrial tissues in the secretory
phase and the decidua. J Clin Endocrinol Metab 86,
5609-14, 2001.
27. Kugu K, Momoeda M, Sharma SS, Osuga Y, Fujiwara
T, Okagaki R, Fukushima H, Yano T, Tsutsumi O
and Taketani Y. Is an elevation in basal follicle-stimulating hormone levels in unexplained infertility predictive of fecundity regardless of age: Endocr J 48,
711-5, 2001.
28. Marumo G, Kozuma S, Ohyu J, Hamai Y, Machida Y,
Kobayashi K, Ryo E, Unno N, Fujii T, Baba K, Okai T,
Takashima S and Taketani Y. Generation of periven-

200

tricular leukomalacia by repeated umbilical cord
occlusion in near-term fetal sheep and its possible
pathogenetical mechanisms. Biol Neonate 79, 39-45,
2001.
29. Minaguchi T, Yoshikawa H, Oda K, Ishino T, Yasugi
T, Onda T, Nakagawa S, Matsumoto K, Kawana K
and Taketani Y. PTEN mutation located only outside
exons 5, 6, and 7 is an independent predictor offavorable survival in endometrial carcinomas. Clin
Cancer Res 7, 2636-42, 2001.
30. Osuga Y, Koga K, Tsutsumi O, Yano T, Kugu K,
Momoeda M, Okagaki R, Suenaga A, Fujiwara T,
Fujimoto A, Matsumi H, Hiroi H and Taketani Y.
Evidence for the presence of keratinocyte growth factor (KGF) in human ovarian follicles. Endocr J 48,
161-6, 2001.
31. Ryo E, Shiotsu H, Takai Y, Tsutsumi O, Okai T,
Taketani Y and Takeuchi Y. Effects of pulsed ultrasound on development and glucose uptake of preimplantation mouse embryos. Ultrasound Med Biol 27,
999-1002, 2001.
32. Ryo E, Yorinaga Y, Nagasaka T, Yoshikawa H and
Taketani Y. Tumor cell spillage to the vaginal cavity
and vaginal stump during the surgery of endometrial
carcinoma. Acta Obstet Gynecol Scand 80, 364-7,
2001.
33. Tsuchiya F, Ikeda K, Tsutsumi O, Hiroi H, Momoeda
M, Taketani Y, Muramatsu M and Inoue S.
Molecular cloning and characterization of mouse
EBAG9, homolog of a human cancer associated surface antigen: expression and regulation by estrogen.
Biochem Biophys Res Commun 284, 2-10, 2001.
34. Uemura H, Unno N, Osuga Y, Momoeda M, Ando K,
Marumo G, Kikuchi A, Fujii T, Kozuma S and
Taketani Y. A movable gestational sac in association
with a myometrial defect. Ultrasound Obstet
Gynecol 18, 675-7, 2001.
35. Xin CY, Matsumoto K, Yoshikawa H, Yasugi T, Onda
T, Nakagawa S, Yamada M, Nozawa S, Sekiya S,
Hirai Y, Shiromizu K, Fujii T and Taketani Y.
Analysis of E6 variants of human papillomavirus
type 33, 52 and 58 in Japanese women with cervicalintraepithelial neoplasia/cervical cancer in relation totheir oncogenic potential. Cancer Lett 170, 19-24,
2001.
36. Yoshida SH, Unno N, Kagawa H, Shinozuka N,
Kozuma S and Taketani Y. Sonographic determination of fetal size from 20 weeks of gestation onward
correlates with birth weight. J Obstet Gynaecol Res
27, 205-11, 2001.
37. Yoshino O, Osuga Y, Koga K, Tsutsumi O, Yano T,
Fujii T, Kugu K, Momoeda M, Fujiwara T, Tomita K
and Taketani Y. Evidence for the expression of interleukin (IL)-18, IL-18 receptor and IL-18 binding protein in the human endometrium. Mol Hum Reprod 7,
649-54, 2001.
38. Fujimoto A, Osuga Y, Yano T, Kusumi M, Kurosawa
T, Fujii T and Taketani Y. Ovarian hyperstimulation
syndrome complicated by peritonitis due to perforated appendicitis. Hum Reprod 17, 966-7, 2002.

39. Hashimoto K, Kato K, Imamura K, Kishimoto A,
Yoshikawa H, Taketani Y and Esumi H. 5-amino-4imidazolecarboxamide riboside confers strong tolerance to glucose starvation in a 5’-AMP-activated protein kinase-dependent fashion. Biochem Biophys Res
Commun 290, 263-7, 2002.
40. Hasumi Y, Mizukami H, Urabe M, Kohno T,
Takeuchi K, Kume A, Momoeda M, Yoshikawa H,
Tsuruo T, Shibuya M, Taketani Y and Ozawa K.
Soluble FLT-1 expression suppresses carcinomatous
ascites in nude mice bearing ovarian cancer. Cancer
Res 62, 2019-23, 2002.
41. Hiroi H, Osuga Y, Tarumoto Y, Shimokama T, Yano
T, Yokota H and Taketani Y. A case of estrogen-producing Brenner tumor with a stromal component asa potential source for estrogen. Oncology 63, 201-4,
2002.
42. Horie K, Tomida A, Sugimoto Y, Yasugi T,
Yoshikawa H, Taketani Y and Tsuruo T. SUMO-1
conjugation to intact DNA topoisomerase I amplifies
cleavable complex formation induced by camptothecin. Oncogene 21, 7913-22, 2002.
43. Kawana K, Yasugi T, Kanda T, Kawana Y, Hirai Y,
Yoshikawa H and Taketani Y. Neutralizing antibodies against oncogenic human papillomavirus as a possible determinant of the fate of low-grade cervicalintraepithelial neoplasia. Biochem Biophys Res
Commun 296, 102-5, 2002.
44. Matsumoto K, Yoshikawa H, Yasugi T, Onda T,
Nakagawa S, Yamada M, Kawana K, Minaguchi T,
Oda K, Hasumi Y and Taketani Y. Distinct lymphatic
spread of endometrial carcinoma in comparison withcervical and ovarian carcinomas. Cancer Lett 180, 839, 2002.
45. Momoeda M, Taketani Y, Terakawa N, Hoshiai H,
Tanaka K, Tsutsumi O, Osuga Y, Maruyama M,
Harada T, Obata K and Hayashi K. Is endometriosis
really associated with pain? Gynecol Obstet Invest 54
Suppl 1, 18-23, 2002.
46. Okada S, Tsuda H, Takarabe T, Yoshikawa H,
Taketani Y and Hirohashi S. Allelotype Analysis of
Common Epithelial Ovarian Cancers with Special
Reference to Comparison between Clear Cell
Adenocarcinoma with Other Histological Types. Jpn
J Cancer Res 93, 798-806, 2002.
47. Osuga Y, Yano T, Kugu K, Koga K, Fukuoka K,
Matsumi H, Wada O, Kikuchi A, Tsutsumi O and
Taketani Y. Effects of gonadotropin-releasing hormone analog treatment on skin condition. Gynecol
Endocrinol 16, 57-61, 2002.
48. Tang X, Yano T, Osuga Y, Matsumi H, Yano N, Xu J,
Wada O, Koga K, Kugu K, Tsutsumi O, Schally AV
and Taketani Y. Cellular mechanisms of growth inhibition of human epithelial ovarian cancer cell line by
LH-releasing hormone antagonist Cetrorelix. J Clin
Endocrinol Metab 87, 3721-7, 2002.
49. Xu J, Osuga Y, Yano T, Morita Y, Tang X, Fujiwara
T, Takai Y, Matsumi H, Koga K, Taketani Y and
Tsutsumi O. Bisphenol A induces apoptosis and G2-

to-M arrest of ovarian granulosa cells. Biochem
Biophys Res Commun 292, 456-62, 2002.
50. Zhang SQ, Kozuma S, Tanaka M, Ling D, Kitano Y,
Fujii T, Baba K and Taketani Y. Studies on fetal forelimb movements by using a wrist actigraph in sheep.
Psychiatry Clin Neurosci 56, 283-4, 2002.

201

Department of
Gynecologic Surgery
Outline and Research Objectives
Our department was established for development of effective and safe gynecologic surgery on the basis of knowledge in reproductive medicine and gynecologic oncology.

Faculties and Students
Professor and Chair: Osamu Tsutsumi, MD, PhD (from2001)
Associate Professor: Tetsu Yano
Lecturer:
Osamu Nishii
Tomoyuki Fujii
Associate: ................................2
Postdoctoral Fellow: ...............2
Graduate Student:...................2
Research Student: ...................5
Secretary: ................................2

Past Research and Major Accomplishments
Professor Tsutsumi has been very actively working
on both clinical and basic research in our department
even before becoming Professor in 2001. In the clinical field, he conducted endoscopic surgery as the first
person in gynecology. As for the basic research, many
achievements were left so far in the embryogenesis,
the genetics and the reproductive endocrinology.

Current Research
For the benign gynecological disorders, we havebeen constantly trying to minimize surgical invasion
to patients as much as possible. With both of wellequipped instruments and well-trained expertise,
more than 90% of surgeries for benign gynecologicaldisorders are operated endoscopically, which make atotal of 300 cases per year. On the other hand, various
projects concerning treatment of gynecological malignancies and sexually transmitted viral infections areon-going. To improve the prognosis of gynecologicalmalignancy, we are conducting the studies, including
1) Prognostic significance of pelvic and paraaortic
lumphnode metastasis in the uterine cervical cancer,2) application of chemotherapy (BLM + VCR + MMC+ CDDP) for advanced or recurrent cervical cancer, 3)
incidence and distribution of PLN and PALN metastasis in endometrial cancer, 4) relationship between thepresence of estrogen and progesterone receptor andclinical course in endometrial cancer, 5) development202

of optimal surgical procedure for advanced ovariancancer, 6) impacts of aggressive debulking (including
bowel resection) on the prognosis of advanced ovariancancer, 7) analysis of long term prognosis of ovariancancer patients treated with paclitaxel containing
chemotherapy, 8) analysis of adverse effects of CDDPbased chemotherapy, focused on nephrotoxicity andneurotoxicity, 9) effect of intermittent consolidation
chemotherapy for advanced ovarian cancer, 10) evaluation of sensitivity and specificity of serum CA125level assay and ultrasonographic follow-up for diagnosis of recurrent ovarian cancer, 11) trans-arterial
chemotherapy for parenchymal liver metastasis ofovarian cancer, 12) estimation of effects of a combination chemotherapy (hydroxyurea + etoposide + DTIC)
for uterine sarcomas, 13) cohort study of low-grade
CIN.
In basic research section, the studies are on goingon the effect of endocrine disrupters on the reproductive system, embryogenesis and oncogenesis.

Future Prospects
As well as proceeding the studies mentioned
above, we are starting the project on developing vaccine against HPVs, using neutralizing epitope of HPV16 capsid proteins L1 and L2 (L1/L2 capsids).

Research Grants
1. Grant-in-Aid for Core Research for Evolutional
Science and Technology, Japan Science andThechnology Corporation: “Effect of endocrine disrupters on reproductive functions ” (1998-2003)
2. Grant-in-Aid for Scientific Research, Japan Society
for the Promotion of Science: Scientific Research (B)“Effect of endocrine disrupters on reproductive functions” (1999-2001)
3. Grant-in-Aid for Scientific Research, Japan Society
for the Promotion of Science: Scientific Research (B)“Effect of endocrine disrupters on reproductive functions” (2002-2004)
4. Grant-in-Aid for Scientific Research, Ministry of
Health, Labor and Welfare: “Effect of endocrine disrupters on reproductive health” (1998-2000)

5. Grant-in-Aid for Scientific Research, Ministry of
Health, Labor and Welfare: “Effect of endocrine disrupters on reproductive health” (2001-2002)

Select Publications
1. Tsutsumi O, Kurachi H and Oka T. A physiologicalrole of epidermal growth factor in male reproductivefunction. Science 233, 975-7, 1986.
2. Tsutsumi O, Kubota Y and Oka T. Effect of sialoadenectomy, treatment with epidermal growth factor(EGF) antiserum and replacement of EGF on the epidermis in mice. J Endocrinol 113, 193-7, 1987.
3. Tsutsumi O and Oka T. Epidermal growth factor
deficiency during pregnancy causes abortion in mice.
Am J Obstet Gynecol 156, 241-4, 1987.
4. Tsutsumi O, Tsutsumi A and Oka T. Importance of
epidermal growth factor in implantation and growthof mouse mammary tumor in female nude mice.
Cancer Res 47, 4651-3, 1987.
5. Tsutsumi O, Tsutsumi A and Oka T. A possible
physiological role of milk epidermal growth factor inneonatal eyelid opening. Am J Physiol 252, R376-9,
1987.
6. Tsutsumi O, Tsutsumi A and Oka T. Epidermal
growth factor-like, corneal wound healing substancein mouse tears. J Clin Invest 81, 1067-71, 1988.
7. Tsutsumi O, Yano T, Satoh K, Mizuno M and Kato T.
Studies of hexokinase activity in human and mouse
oocyte. Am J Obstet Gynecol 162, 1301-4, 1990.
8. Tsutsumi O, Satoh K, Taketani Y and Kato T.
Determination of enzyme activities of energy metabolism in the maturing rat oocyte. Mol Reprod Dev
33, 333-7, 1992.
9. Sadatsuki M, Tsutsumi O, Yamada R, Muramatsu M
and Taketani Y. Local regulatory effects of activin A
and follistatin on meiotic maturation of rat oocytes.
Biochem Biophys Res Commun 196, 388-95, 1993.
10. Sadatsuki M, Tsutsumi O, Sakai R, Eto Y, Hayashi N
and Taketani Y. Presence and possible function ofactivin-like substance in human follicular fluid. Hum
Reprod 8, 1392-5, 1993.
11. Tsutsumi O, Iida T, Hakuno N, Sadatsuki M, Okai T,
Taketani Y, Nagafuchi S and Nakahori Y. Y chromosome analysis and laparoscopic surgery in XY puregonadal dysgenesis: a case report and a review of literature. Asia Oceania J Obstet Gynaecol 19, 95-9,
1993.
12. Tsutsumi O, Taketani Y and Oka T. The uterine
growth-promoting action of epidermal growth factor
and its function in the fertility of mice. J Endocrinol
138, 437-44, 1993.
13. Tsutsumi O, Taketani Y and Oka T. Evidence for the
involvement of epidermal growth factor in fertility
decline in aging female mice. Horm Res 39 Suppl 1,
32-6, 1993.
14. Ayabe T, Tsutsumi O and Taketani Y. Hexokinase
activity in mouse embryos developed in vivo and invitro. Hum Reprod 9, 347-51, 1994.

15. Morita Y, Tsutsumi O and Taketani Y. In vitro treatment of embryos with epidermal growth factor
improves viability and increases the implantation
rate of blastocysts transferred to recipient mice. Am
J Obstet Gynecol 171, 406-9, 1994.
16. Tsutsumi O, Iida T, Taketani Y, Sugase M, Nakahori
Y and Nakagome Y. Intact sex determining region Y(SRY) in a patient with XY pure gonadal dysgenesis
and a twin brother. Endocr J 41, 281-5, 1994.
17. Tsutsumi O, Yano T and Taketani Y.
Phosphofructokinase activity as a measure of maturation of rat oocytes developed in vivo and in vitro.
Horm Res 41 Suppl 1, 63-7, 1994.
18. Morita Y, Tsutsumi O and Taketani Y. Successfultreatment of catamenial pneumothorax with danazol. Int J Gynaecol Obstet 51, 263-4, 1995.
19. Tsutsumi O, Iida T and Taketani Y. Laparoscopic
surgery and DNA analysis in patients with XY puregonadal dysgenesis. J Obstet Gynaecol 21, 67-74,
1995.
20. Morita Y, Tsutsumi O, Kuramochi K, Momoeda M,
Yoshikawa H and Taketani Y. Successful laparoscopic management of primary abdominal pregnancy.
Hum Reprod 11, 2546-7, 1996.
21. Takai Y, Ogawara M, Tomono Y, Moritoh C, ImajohOhmi S, Tsutsumi O, Taketani Y and Inagaki M.
Mitosis-specific phosphorylation of vimentin by protein kinase C coupled with reorganization of intracellular membranes. J Cell Biol 133, 141-9, 1996.
22. Tsutsumi O, Iida T, Nakahori Y and Taketani Y.
Analysis of the testis-determining gene SRY in
patients with XY gonadal dysgenesis. Horm Res 46
Suppl 1, 6-10, 1996.
23. Tsutsumi O and Yoshimura Y. Sex differentiationand ovarian function. An overview. Horm Res 46
Suppl 1, 1-5, 1996.
24. Kawana K, Yoshikawa H, Yokota H, Onda T,
Nakagawa K, Tsutsumi O and Taketani Y. Successfultreatment of brain metastases from ovarian cancer
using gamma-knife radiosurgery. Gynecol Oncol 65,
357-9, 1997.
25. Morita Y, Tsutsumi O and Taketani Y. Successful
hormonal treatment of pulmonary parenchymal
endometriosis. Int J Gynaecol Obstet 59, 61-3, 1997.
26. Morita Y, Tsutsumi O, Momoeda M and Taketani Y.
Cornual pregnancy successfully treated laparoscopically with fibrin glue hemostasis. Obstet Gynecol 90,
685-7, 1997.
27. Sadatsuki M, Kaneko M and Tsutsumi O. Expectant
management and methotrexate treatment of persistent ectopic pregnancy following laparoscopic salpingectomy. Int J Gynaecol Obstet 59, 49-51, 1997.
28. Tsutsumi O, Ando K and Momoeda M. Ruptured
isthmal pregnancy following laparoscopic salpingostomy in the ipsilateral tube. Int J Gynaecol Obstet
57, 187-9, 1997.
29. Uechi H, Tsutsumi O, Morita Y and Taketani Y.
Cryopreservation of mouse embryos affects laterembryonic development possibly through reduced203expression of the glucose transporter GLUT1. Mol
Reprod Dev 48, 496-500, 1997.
30. Takai Y, Tsutsumi O, Harada I, Fujii T, Kashima T,
Kobayashi K, Toda T and Taketani Y. Prenatal diagnosis of Fukuyama-type congenital muscular dystrophy by microsatellite analysis. Hum Reprod 13, 3203, 1998.
31. Tsutsumi O, Uechi H, Sone H, Yonemoto J, Takai Y,
Momoeda M, Tohyama C, Hashimoto S, Morita M
and Taketani Y. Presence of dioxins in human follicular fluid: their possible stage-specific action on thedevelopment of preimplantation mouse embryos.
Biochem Biophys Res Commun 250, 498-501, 1998.
32. Igarashi T, Osuga U, Tsutsumi O, Momoeda M,
Ando K, Matsumi H, Takai Y, Okagaki R, Hiroi H,
Fujiwara O, Yano T and Taketani Y. Expression of
Ah receptor and dioxin-related genes in human uterine endometrium in women with or without
endometriosis. Endocr J 46, 765-72, 1999.
33. Okagaki R, Osuga Y, Momoeda M, Tsutsumi O and
Taketani Y. Laparoscopic findings after ultrasoundguided transvaginal ethanol sclerotherapy for ovarian
endometrial cyst. Hum Reprod 14, 270, 1999.
34. Takai Y, Tsutsumi O, Momoeda M, Osuga Y,
Sadatsuki M, Kaibara M and Taketani Y. Non-functioning pituitary tumour after long-term treatment
with gonadotrophin-releasing hormone agonists in apatient with vaginal agenesis who underwent neovaginoplasty and cauterization of endometriosis
under laparoscopy. Hum Reprod 14, 2661-4, 1999.
35. Uechi H, Tsutsumi O, Morita Y, Takai Y and
Taketani Y. Comparison of the effects of controlledrate cryopreservation and vitrification on 2-cellmouse embryos and their subsequent development.
Hum Reprod 14, 2827-32, 1999.
36. Maruyama M, Osuga Y, Momoeda M, Yano T,
Tsutsumi O and Taketani Y. Pregnancy rates after
laparoscopic treatment. Differences related to tubal
status and presence of endometriosis. J Reprod Med
45, 89-93, 2000.
37. Morita Y, Nishii O, Kido M and Tsutsumi O.
Parvovirus infection after laparoscopic hysterectomy
using fibrin glue hemostasis. Obstet Gynecol 95,
1026, 2000.
38. Takai Y, Tsutsumi O, Ikezuki Y, Hiroi H, Osuga Y,
Momoeda M, Yano T and Taketani Y. Estrogen
receptor-mediated effects of a xenoestrogen, bisphenol A, on preimplantation mouse embryos. Biochem
Biophys Res Commun 270, 918-21, 2000.
39. Takai Y, Tsutsumi O, Harada I, Morita Y, Momoeda
M, Fukushima Y and Taketani Y. A case of XY puregonadal dysgenesis with 46,XYp-/47,XXYp- karyotype whose gonadoblastoma was removed laparoscopically. Gynecol Obstet Invest 50, 166-9, 2000.
40. Takeuchi T and Tsutsumi O. Basal leptin concentrations in women with normal and dysfunctional ovarian conditions. Int J Gynaecol Obstet 69, 127-33,
2000.
41. Tsutsumi O, Momoeda M, Takai Y, Ono M and
Taketani Y. Breast-fed infants, possibly exposed to204

dioxins in milk, have unexpectedly lower incidence
of endometriosis in adult life. Int J Gynaecol Obstet
68, 151-3, 2000.
42. Fujimoto A, Osuga Y, Tsutsumi O, Fujii T, Okagaki R
and Taketani Y. Successful laparoscopic treatment of
ileo-cecal endometriosis producing bowel obstruction. J Obstet Gynaecol Res 27, 221-3, 2001.
43. Morita Y, Tsutsumi O and Taketani Y. Regulatory
mechanisms of female germ cell apoptosis during
embryonic development. Endocr J 48, 289-301, 2001.
44. Osuga Y, Tsutsumi O, Fujiwara T, Kugu K, Fujimoto
A and Taketani Y. Usefulness of long-jaw forceps inlaparoscopic cornual resection of interstitial pregnancies. J Am Assoc Gynecol Laparosc 8, 429-32, 2001.
45. Takai Y, Tsutsumi O, Ikezuki Y, Kamei Y, Osuga Y,
Yano T and Taketan Y. Preimplantation exposure tobisphenol A advances postnatal development.
Reprod Toxicol 15, 71-4, 2001.
46. Ikezuki Y, Tsutsumi O, Takai Y, Kamei Y and
Taketani Y. Determination of bisphenol A concentrations in human biological fluids reveals significant
early prenatal exposure. Hum Reprod 17, 2839-41,
2002.
47. Kurosawa T, Hiroi H, Tsutsumi O, Ishikawa T,
Osuga Y, Fujiwara T, Inoue S, Muramatsu M,
Momoeda M and Taketani Y. The activity of bisphenol A depends on both the estrogen receptor subtype
and the cell type. Endocr J 49, 465-71, 2002.
48. Nakagawa S, Koga K, Kugu K, Tsutsumi O and
Taketani Y. The evaluation of the sentinel node successfully conducted in a case of malignant melanomaof the vagina. Gynecol Oncol 86, 387-9, 2002.49.
Osuga Y, Koga K, Tsutsumi O, Yano T, Maruyama M,
Kugu K, Momoeda M and
49. Osuga Y, Koga K, Tsutsumi O, Yano T, Maruyama M,
Kugu K, Momoeda M and Taketani Y. Role oflaparoscopy in the treatment of endometriosis-associated infertility. Gynecol Obstet Invest 53 Suppl 1,
33-9, 2002.
50. Takeuchi T and Tsutsumi O. Serum bisphenol a concentrations showed gender differences, possibly
linked to androgen levels. Biochem Biophys Res
Commun 291, 76-8, 2002.

Department of Pediatrics /Developmental Pediatrics
Outline and Research Objectives
Department of Pediatrics was founded in 1889 and continued its activity for 113 years.
This is the oldest Pediatrics and has been the leader of Pediatrics in Japan. Many pediatricians received the training in our department and went to all over and out of Japan andworked as a leader of pediatrician in a lot of Japanese Medical School in the University, central and local hospitals and clinics.
The health problems of children and youth have changed historically in Japan. In thelate 19th century, one of five children died as infectious diseases such as dysentery, pneumonia, diphtheria, and whooping cough. Now, few children die due to the diseases. The leading causes of death for children are perinatal problems for babies, injuries, congenital anomalies, malignant neoplasms, and congenital heart diseases. Many of these diseases havegenetic background in their pathogenesis. Thus, the research objectives have greatlychanged. Now, they are to clarify the molecular pathogenesis of these difficult diseases andto find safe and effective therapies for them and to find the way to promote the body andheart health of children in our Pediatrics.
Pediatrics covers all the issues in children and youth. Thus, our department studies
almost all the issues about children and youth. Now, we have six active study groups suchas Nephrology, Hematology and Oncology, Cardiology, Immunology and Allergy,
Endocrinology and Metabolism, and Neurology. The main subjects of the research are 1)
Identification of the molecular pathogenesis of renal tubular disorders, 2) Clarification andcharacterization of the genes which cause leukemia and neuroblastoma, 3) Role of hemodynamic factors on pulmonary hypertension complicated with congenital heart diseases, 4)
Epidemiological survey and nationwide registry of primary immunodeficiency diseases andthe efficacy of immunotherapy for cancer using peripheral dendritic cells, 5) Molecular andfunctional analysis of the genes for inherited endocrine and metabolic diseases, and 6)
Pathogenesis of peroxisome biogenesis disorders. The details of the research projects aredescribed bellow.
We have been the local registration center for children’s cancer in Kantou-district. This
helps and promotes the basic research in the fields of Hematology and Oncology in ourPediatrics.

Faculties and Students
Professor and Chair Takashi Igarashi, M.D., Ph.D. (2000-)
Associate Professors Tsutomu Iwata, M.D., Ph.D.
Lecturers
Yasuhide Hayashi, M.D., Ph.D.
Masaru Takamizawa, M.D., Ph.D.
Hitoshi Kato, M.D., Ph.D.
Takashi Sekine, M.D., Ph.D.
Yoichi Sakakihara, M.D., Ph.D.
Associates .............................15
Postdoctoral Fellow ................4
Graduate student ..................18
Research student.....................3
Secretary .................................3

Past Research and Major Accomplishments
Nephrology
1) Molecular pathogenesis of Dent’s disease, proximal renal tubular acidosis and renal hypouricemia:
feedback from bench to clinics.
2) The mechanisms of post-diarrheal hemolytic uremic syndrome (HUS).
3) Molecular identification of the proximal tubulartransporters that excrete drugs from the kidney.
Hematology and Oncology
1) Identification of HOXD11, HOXA13 and HOXC11
genes as partner genes of NUP98 in t (2;11)-, t(7;11)- and t (11;12)- acute myeloid leukemias.
2) Molecular analysis of tumor suppressor genes inneuroblastoma.

205

Cardiology
1) Role of hemodynamic factors on pulmonary
hypertension complicated with congenital heartdiseases.
2) Development of coronary artery from cardiac neural crest cells.
3) Effect of epoprostenol on severe port pulmonary
hypertension complicated with biliary atresia.
4) Historical review of Kawasaki disease: When
Kawasaki disease started in Japan?
Immunology and Allergy
1) Epidemiological survey and nationwide registry ofprimary immunodeficiency diseases.
2) The efficacy of immunotherapy for cancer using
peripheral dendritic cells.
3) The effective and safe simultaneous administration of several vaccines in a short period of time
before liver transplantation and their administration under immunosuppressive conditions after
transplantation.
4) Prospective study of atopic status in infants of thecohort in Tokyo, Japan.
5) Epidemiology of bronchial asthma among children
in rural and urban area in Bangladesh.
Endocrinology and Metabolism
1) Molecular and functional analysis of the genes forseveral inherited endocrine and metabolic diseases
including the glutamate dehydrogenase gene incongenital hyperinsulinism hyperammonemia
syndrome, glucose-6-phophatase gene in glycogenstorage disease type 1a, CREB-binding protein in
Rubinstein-Taybi syndrome, hepatocyte nuclear
factor-1b in maturity-onset diabetes of the young
type 5.
Neurology
1) Pathogenesis of peroxisome biogenesis disorders:Altered lipid metabolism in peroxisome-defective
cells.
2) The genetic analysis of Japanese patients withspinal muscular atrophy.
3) The analysis of pathophysiology of epileptic syndrome and higher brain function inchildren by magnetoencephalography.
4) Development of communication tool for patientswith spinal muscular atrophy.

Current Research
Nephrology
1) Molecular and functional analysis of various ion
transporters such as Cl channels, Na +/HCO3 cotransporter, URAT1, barttin, and H+-ATPase.

2062) Molecular and functional analysis of LMX1B in
nail patella syndrome and hepatocyte nuclear factor-1 β in MODY5.
3) The mechanisms of post-diarrheal HUS.
Hematology and Oncology
1) Identification of HOXD11, HOXA13 and HOXC11
genes as partner genes of NUP98 in t (2;11)-, t(7;11)- and t (11;12)- acute myeloid leukemias.
2) Molecular analysis of tumor suppressor genes inneuroblastoma.
Cardiology
1) Role of hemodynamic factors on pulmonary
hypertension complicated with congenital heartdiseases.
2) Development of coronary artery from cardiac neural crest cells.
Immunology and Allergy
1) Epidemiological survey and nationwide registry ofprimary immunodeficiency diseases.
2) The efficacy of immunotherapy for cancer using
peripheral dendritic cells.
3) Prospective study of atopic status in infants of thecohort in Tokyo, Japan.
Epidemiology of bronchial asthma among children
in rural and urban area in Bangladesh.
5) Analysis of the function of basophils in allergic
diseases.
Endocrinology and Metabolism
1) Molecular and functional analysis of the genes forseveral inherited endocrine and metabolic diseases
including the glutamate dehydrogenase gene incongenital hyperinsulinism hyperammonemia
syndrome, glucose-6-phophatase gene in glycogenstorage disease type 1a, CREB-binding protein in
Rubinstein-Taybi syndrome, hepatocyte nuclear
factor-1b in maturity-onset diabetes of the young
type 5.
Neurology
1) Pathogenesis of peroxisome biogenesis disordersAltered lipid metabolism in peroxisome-defective
cells.
2) The analysis of pathophysiology of epileptic syndrome and higher brain function in children bymagnetoencephalography.
3) Development of communication tool for patientswith spinal muscular atrophy.

Future Prospects
Nephrology
1) Molecular and functional analysis of various renaltransporter disorders.
2) Identification of the genes for minimal change
nephrotic syndrome.
Hematology and Oncology
1) Identification and characterization of HOXD11,
HOXA13 and HOXC11 genes as partner genes of
NUP98 in t (2;11)-, t (7;11)- and t (11;12)- acute
myeloid leukemias.
2) Molecular analysis of tumor suppressor genes inneuroblastoma.
Cardiology
1) Role of hemodynamic factors on pulmonary
hypertension complicated with congenital heartdiseases.
2) Development of coronary artery from cardiac neural crest cells.
Immunology and Allergy
1) Epidemiological survey and nationwide registry ofprimary immunodeficiency diseases.
2) The efficacy of immunotherapy for cancer using
peripheral dendritic cells.
3) Prospective study of atopic status in infants of thecohort in Tokyo, Japan.
4) Epidemiology of bronchial asthma among children
in rural and urban area in Bangladesh.
5) Analysis of the function of basophils in allergic
diseases.
Endocrinology and Metabolism
1) Mutational and functional analysis of the genes
for several inherited endocrine and metabolic diseases.
Neurology
1) Pathogenesis of peroxisome biogenesis disordersAltered lipid metabolism in peroxisome-defective
cells.
2) The analysis of higher brain function by magnetoencephalography.
3) Development of communication tool for patientswith spinal muscular atrophy.
Research Grants (5 selections)
1) Igarashi T: Early diagnosis, management and treatment of intractable kidney and urinary tract diseases (Kodomokatei-H13-011). Ministry of Health,
Labor and Welfare, 16,000,000 yen in 2001-2002.

2) Iwata T: The association between Urbanization
and Childhood Asthma in Bangladesh.Nissan
Science Foundation, 4,500,000 yen in 2000-2001,
3,600,000 yen in 2002-2003.
3) Hayashi Y: Analysis of molecular mechanism ofcancer formation due to translocation gene byusing genome information and expression profile(B-2-14370242). Ministry of Education Science,
8,800,000 yen in 2002-2004.
4) Hayashi Y: Gene analysis and diagnosis of
intractable childhood leukemia and secondary
leukemia and their application to the clinics.
Ministry of Health, Labor and Welfare, 7,400,000
yen in 1996-2002.
5) Kato H: Role of hemodynamic factors on pulmonary hypertension complicated with congenital
heart diseases (No. 09470177). Ministry of Health,
Labor and Welfare, 5,300,000 yen in 1999-2001.

Select Publications
Nephrology
1. Sekine T, Cha SH, Hosoyamada M, Kanai Y,
Watanabe N, Huruta Y, Hukuda K, Igarashi T, Endou
H: Cloning, functional characterization and localization of a rat renal Na+-dicarboxylate transporter. Am
J Physiol 275 (Renal Physiol. 44): F298-F305, 1998
2. Igarashi T, Günter W, Sekine T, Inatomi J, Shiraga H,
Takahashi S, Suzuki J, Tsuru N, Yanagihara T,
Shimazu M, Jentsch TJ, Thakker RV: Functional
characterization of renal chloride channel, CLCN5,
mutations associated with Dent’s Japan disease. Kidney
Int 54:1850-1856, 1998
3. Igarashi T, Inatomi J, Sekine T, Cha SH, Kanai Y,
Kunimi M, Tsukamoto K, Satoh H, Shimadzu M,
Tozawa F, Mori T, Shiobara M, Seki G, Endou H:
Mutations in SLC4A4 cause permanent isolated
proximal renal tubular acidosis with ocular abnormalities. Nature Genet 23:264-266, 1999
4. Igarashi T, Inatomi J, Wake A, Takamizawa M,
Katayama H, Iwata T: Failure of pre-diarrheal antibiotics to prevent hemolytic uremic syndrome in serologically-proven Escherichia coli 0157:H7 gastrointestinal infection. J Pediatr 135: 768-769, 1999
5. Igarashi T, Inatomi J, Ohara T, Kuwahara T,
Shimadzu M, Thakker RV: Clinical and genetic studies of CLCN5 mutations in Japanese families with
Dent’s disease. Kidney Int 58: 520-527, 2000
6. Igarashi T, Inatomi J, Sekine T, Seki G, Shimadzu M,
Tozawa F, Takeshima Y, Takumi T, Takahashi T,
Yoshikawa N, Nakamura H, Endou H: Novel nonsense mutation in the Na+/HCO3- cotransporter gene
(SLC4A4) in a patient with permanent isolated proximal renal tubular acidosis and bilateral glaucoma. J
Am Soc Nephrol 12: 713-718, 2001
7. Usui T, Hara M, Satoh H, Moriyama N, Kagaya H,
Amano S, Oshika T, Ishii Y, Ibaraki N, Hara C,
Kunimi M, Noiri E, Tsukamoto K, Inatomi J,

207

Kawakami H, Endou H, Igarashi T, Goto A, Fujita T,
Araie M, Seki G: Molecular basis of ocular abnormalities associated with proximal renal tubular acidosis. J
Clin Invest 108: 107-115, 2001
8. Kuwahara M, Iwai K, Ooeda T, Igarashi T, Ogawa E,
Katsushima Y, Shinbo I, Uchida S, Terada Y,
Fujiwara TM, Arthus MF, Lonergan M, Bicher DG,
Marumo F: Three families with autosomal dominant
nephrogenic diabetes insipidus caused by aquaporin2 mutations in the C-terminus. Am J Human
Genetics 69: 738-748, 2001
9. Norden AGW, Lapsley M, Christensen EI, Igarashi T,
Matsuyama T, Shiraga H, Kelleher CL, Lee P,
Scheinman SJ, Thakker RV, Unwin RJ, Verroust P,
Wrong O, Moestrup S: Urinary megalin deficiency
implicates abnormal tubular endocytic receptor trafficking in the Fanconi syndrome. J Am Soc Nephrol
13: 125-133, 2002
10. Chandler WL, Jelacic S, Boster D, Ciol M, Watkins S,
Igarashi T, Tarr PI: Prothrombotic coagulation abnormalities preceding the hemolytic uremic syndrome.
New Eng J Med 246: 23-32, 2002
11. Enomoto A, Kimura H, Chairoungdua A, Shigeta Y,
Jutabha P, Cha SH, Hosoyamada M, Takeda M,
Sekine T, Igarashi T, Matsuo H, Kikuchi Y, Oda T,
Ichida K, Hosoya T, Shimokata K, Niwa T, Kanai Y,
Endou H: Molecular identification of arenal/urate/anion exchanger that regulates bloodurate levels. Nature 417: 447-452, 2002

Hematology and Oncology
12. Futatani T, Miyawaki T, Tsukada S, Hashimoto S,
Kunikata T, Arai S, Kurimoto M, Niida Y, Matsuoka
H, Sakiyama Y, Iwata T, Tsuchiya S, Tatsuzawa O,
Yoshizaki K, Kishimoto T: Deficient expression of
Bruton’s tyrosine kinase in monocytes from X-linked
agammaglobulinemia as evaluated by a flow cytometric analysis and its clinical application to carrier
detection. Blood 15:595-602, 1998
13. Ishiguro A, Ishikita T, Shimbo T, Matsubara K, Baba
K, Hayashi Y, Naritaka S, Nakahata T: Elevation ofserum thrombopoietin precedes thrombocytosis in
Kawasaki disease. Thromb Haemost 79:1096-1100,
1998
14. Kitabayashi I, Ida K, Morohoshi F, Yokoyama A,
Mitsuhashi N, Shimizu K, NomuraN, Hayashi Y,
Ohki M: The AML1-MTG8 leukemic fusion protein
forms a complex with A novel member of the MTG8
(ETO/CDR) family, MTGR1. Mol Cell Biol 18:846858, 1998
15. Miyagawa K, Kent J, Moore A, Charlieu JP, Little
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Department of PediatricSurgery / Department ofPediatric Oncology
Outline and Research Objectives
The history of Department of Pediatric Surgery is not long in University of Tokyo. It firststarted as a clinical department of University of Tokyo Hospital in 1971. In 1989 it became adepartment in The Faculty of Medicine and then transferred to The Graduate School of
Medicine as a part of the change of the system. So the duration of research in our department is not long. Also we have only few staff in our department. However, we have continued research very intensively by collaboration with other departments in The Graduate
School of Medicine, University of Tokyo, and other institutes in Japan and abroad.
Our main themes of research are in three areas, namely transplantation medicine, developmental biology of congenital anomalies and fetal surgery, and immunology of intestine.
Department of Pediatric Oncology was separated from Department of Pediatric Surgery
when the system changed, and Graduate School of Medicine became the major. Before that,studies concerning pediatric oncology was done in the Department of Pediatric Surgery andalso in the Department of Pediatrics. Now as a system, Department of Pediatrics is separated from the Department of Pediatric Oncology, the group of pediatric oncology in the
Department of Pediatrics actually works with the Department of Pediatric Oncology, andconstitutes a part of the department.
Pediatric malignancies are very special and consist only a very small part of all the malignancies considering the number of patients. However, because of their peculiarities, they havesome privileges and advantages. For example, about the theory of oncogenesis, famous “twohit theory” by Knudson was first proposed for pediatric malignancies, retinoblastoma andnephroblastoma. It is because that the mechanism of oncogenesis in these tumors is very simple compared to that of adult malignancies. Also the “multi-disciplinary treatment” of malignant diseases first successfully introduced to malignant solid tumors in infancy and children.
So the study of pediatric oncology is very important and it has possibility of break-through inthe study of oncology general. Now we mainly study oncogenes related to pediatric malignancies especially neuroblastoma, and immuno-therapy of cancer using dendritic cells.

Faculties and Students
Professor and Chair

Kohei Hashizume, M.D,,Ph.D..
(since 1997)
Lecturer
Yutaka Kanamori, M.D.,Ph.D..
(Yasuhide Hayashi, M.D.,Ph.D.)
Associates ...............................3
Graduate Students ..................4
Secretary .................................2

Past Research and Major Accomplishments
In the area of transplantation medicine, we started
clinical living-related liver transplantation with the
Department of Artifitial Organ and Transplantationand have operated about 100 pediatric cases.
Associate Professor Kawarasaki in our department
who was in charge in liver transplantation moved toJichi Medical School in 2001. So we stopped clinicalliver transplantation in pediatric cases. But we continue research in this area now. We studied immunological tolerance using rat heart transplantation model,
and found that donor specific transfusion induce
immunological tolerance. Then we analyzed rat serum
after donor specific transfusion using liquid chromatography. We found three kinds of proteins thathave a function of immunosuppression. Especially oneof those 3 protein, MAY-1 is a completely new proteinwith 234 amino acid and has a possibility of usage asa immunosuppression drug in clinical transplantation.
We studied small bowel transplantation using amodel of small bowel transplantation with a free graftof newborn intestine into the recipient’s omentum. In
this model we showed good neovascularization andhistologically mature intestine after the transplantation. We also showed the near normal motility andabsorption in this transplanted intestine. In shortboweled rats, transplantation of newborn intestine211

could restore the weight loss and could save the livesof these short-boweled rats. Also cryopreserved newborn intestine could function as good as fresh intestine after transplantation.
We also studied warm ischemia-reperfusion injury
of the liver in rat model. We showed that pretreatment of the liver with cyclosporine reduce damage tothe liver, but pretreatment with FTY720 rather prolong damage to the liver. So pretreatment withimmuno-suppressive drugs of the liver does not universally beneficial for the prevention of warm
ischemia-reperfusion injury.
In the area of developmental biology of congenital
anomalies and fetal surgery, we very intensively studied congenital diaphragmatic hernia (CDH) using rat
model induced by a drug nitrofen. CDH is one of themost difficult to treat congenital anomalies with highmortality. Until recently its experimental study wasdone using only sheep model that is made by fetal surgery. The problem is that the fetal surgery can be done
only in late phase of pregnancy, many features of thismodel is quite different from that of human CDH. We
noticed rat CDH induced by administration of nitrofen
to pregnant rat. The CDH by this method develop atvery early stage of fetal development and very similar
to human CDH. First we showed clearly that hypoplasia of the lungs develop by CDH, and spreading of
sufactant phospholipids in the alveolar cells to thelumen of alveoli is strongly impaired in fetuses with
CDH. Then we studied the effects of fetal surgery tolung hypoplasia in CDH. We developed experimental
model of tracheal ligation in rat CDH model. Using
this model, we showed that tracheal ligation can overcome the hypoplasia of the lungs induced by CDH. By
ligation of the trachea the lungs grow very rapidly, andthe weight, contents of protein and DNA of the lungs,
and histological structure normalize. By this experiment we clearly showed that tracheal ligation canovercome the hypoplasia of the lungs in CDH, andthat this simple fetal surgery can same the lives ofbabies with CDH and severe hypoplasia of the lungs
whom it is very difficult to treat after birth.
We also developed another new model of congenital
anomaly in a species of amphibian, Cynopus
pyrrhogaster. We succeeded to produce split cord malformation in Cynopus embryos experimentally incising apart of neural plate and notochord. With this procedure,
fistulae developed between endoderm and ectoderm,
and in some cases endodermal or neural cysts developed.
This experiment shed light to the cause of split notochord syndrome and also duplication of intestine.
In the area of immunology of intestine, Kanamori
first described novel lympoid tissues, cryptopatch inmurine intestinal mucosa in 1996. Then we studied
human intestinal mucosa for the existence of lymphoid tissues corresponding to the cryptopatch in212

mouse. We could not show the existence of such lymphoid tissues in human. So we turned to study thefunctions of already known lymphoid tissues inhuman if they have functions corresponding to cryptopatch in mouse and compensate the absence ofthat. Using surgical specimens of jejunum of infants,
we looked for lymphoid tissues in the mucosa of thejejunum, and examined the distribution of these tissues. We found Peyer’s patches and isolated lymphoidfollicles. Peyer’s patches distributed in antimesentericarea, but isolated lymphoid follicles distributed irregularly all through the surface of the mucosa, and inthese isolated lymphoid follicles CD19 positive andCD3 positive lympoid cells are distributed.
We introduced new synbiotics therapy to various
intestinal conditions in children, namely short bowel
syndrome, inflammatory bowel diseases, and pseudoHirschsprung disease. We administered two types ofprobiotics (Bifidobacterium brevs and Lactobacillus
casei) together with prebiotic (galactooligosaccarides)
to these patients. After the introduction of this therapy, the patients’ clinical conditions dramatically
improved. Feces became more solid and less foulsmelling, the patients got weight, abnormal dilatationof the intestine improved, and frequency of bacterialtranslocation decreased. We also showed change ofbacterial flora of feces. Those two types of probiotics
became the main flora, and the number of pathogenic
bacteria dramatically decreased.
In the area of pediatric oncology, we started ourstudy of pediatric oncology in the field of histochemistry. We studied expression of multi-drug resistance
related protein, P-glycoprotein in neuroblastoma cells.
Surprisingly, it was shown that high expression of Pglycoprotein was related to better prognosis ofpatients. It looks paradoxical, but we also found innormal adrenal medulla cells, P-glycoprotein isexpressed. So high expression of P-glycoprotein inneuroblastoma means rather well-differentiation ofthese tumors, and consequently better prognosisi.
Our result was confirmed later by other publications.
Recently we are concentrated in the study of oncogene in pediatric malignancies, especially neuroblastoma and Wilms tumor. It is well known that neuroblastomas whose cells show chromosome 1p deletion
have poor prognosis. We intensively looked for probable oncogene in this region, but it has not yet discovered. There was one great side product of this project.
In this region we found kinesin superfamily motor
protein KIF1B, and it was shown loss-of-function
mutation in the motor domain of the KIF1B genecauses Charcot-Marie-Tooth disease type 2A. We studied expression, LOH, and mutation of p73 gene that is
mapped to 1p36. We found aberrant expression of
p73 is significantly higher in advanced-stages neuroblastoma, but no homozygous deletion of p73 wasfound in any of the sample. So we conclude p73 isless involved in the development but involved in the
progression of neuroblastoma. We also studied othergenes mapped in other chromosomes because otherchromosome anomalies other than 1p deletion are
related to prognosis of neuroblastoma. For example,
17q gain is known to relate to poor prognosis. We
studied survivin mapped to 17q25, which is a memberof inhibitor of apoptosis proteins (IAPs). We foundhigh expression of survivin is a strong prognostic
indicator for the advanced stage neuroblastomas, andthere is a possibility that it is one of the candidate
genes for the 17q gain.
In the study of Wilms tumor, we found a novel
WT1 gene mutation in a patient with Wilms tumor,
male hermaphroditism, but without renal disease.
The mutation was at intron 7 in tumor cells and also
germ line cells. This mutation is thought to produce
truncated WT1 protein, and it is related to the genitourinary anomalies and genesis of Wilms tumor in thispatients. Also the clinical course of this patient was
rather atypical for Wilms tumor in general, and we
think this peculiar mutation should influence thecourse of the disease.
Other than neuroblastoma and Wilms tumor, westudied hepatoblastoma. We examined mutation ofbeta-catenin in the tumors of 68 cases of hepatoblastoma. We found mutation of beta-catenin in 44 (65%)
tumors. In these 44, 9 was point mutation, 35 wasdeletion mutation. We also studied the expression ofbeta-catenin by histochemistry. We found betacatenin is accumulated in the nuclei of tumor cells,
and in these cases with beta-catenin accumulation,
cyclin D1 was also accumulated in the nuclei. We conclude that in hepatoblastoma because of frequent
mutation of beta-catenin gene, beta-catenin protein
accumulates in the nuclei and cause activation of
cyclin D1, that results growth or progression of thetumor. We also found beta-catenin accumulation in
the nuclei of two cases of hepatic adenoma. In one ofthese 2 cases, mutation of beta-catenin gene wasfound. This is the first report of beta-catenin mutationin the case of hepatic adenoma, and we think it shedlight to the genesis of hepatic adenoma and its relation to hepatoblastoma.

Current Research
In the area of transplantation medicine, our main
themes are prevention of ischemia-reperfusion injury
in liver transplantation, and development of safer
method in transplantation of small bowel. In the former theme, we developed partioal liver transplantation model in rat, and studying the effect of gene
introduction to the graft. In the preliminary study, we
got promising effect of gene introduction. In tranplan-tation of small intestine, we have succeeded in thetransplantation of cryopreserved newborn intestine,
but found cryopreservation had no effect on antigenicity of the graft. So we are studying methods toreduce the antigenicity of the intestinal graft toimprove the survival of intestinal transplantation.
In the area of developmental biology of congenital
anomalies and fetal surgery, we are doing experimental fetal surgery of tracheal obstruction using lambs.
We developed new laryngoscope specially designed
for the obstruction of trachea of fetal lambs using balloon. We have successfully introduced balloon into
fetal trachea, and have shown growth of the lungs
after the introduction of balloon.
In the area of immunology of intestine, we are
studying the cells found in isolated lymphoid follicles
in the mucosa of infants, their characters, development, and migration. We are considering of using probiotics in wider varieties of diseases, for example
patients in PICU who receive rather intensive antibiotics therapy and are prone to develop bacterialtranslocation and sepsis.
In Pediatric Oncology Department, we continue thesearch of tumor suppressor gene of neuroblastoma suspected to reside in chromosome 1p. We have established cell line of Wilms tumor with WT1 mutation.
This cell line has morphological features of stromal celland epithelial cell. Using this cell line, we are considering to do a study concerning the factors that determine
the differentiation of the tumor. Wilms tumor shows avery wide variety of histology. In some tumors, histology is almost universally epithelial, but in other tumors,they show stroma-dominant histology with only smallpart of epithelium. Because the histological type relate
to the prognosis of the tumor, we can know the factors
related to prognosis, if we can find out factors that
determine the histological features of the tumor.
Concerning the therapy of malignant tumors inchildren, we are doing a research of immunotherapy
using dendritic cells. Immunotherapy using dendritic
cells is not studied fully in malignant tumors in children. However, it is well known that neuroblastoma
cells show strong tumor-specific antigen, and spontaneous regression of neuroblastoma is well documented. So we think immunotherapy of neuroblastoma is
promising. We found using OK-432, dendritic cells
can be activated, and CTL can be induced. We want totest this method using target of neuroblastoma cells.
Compared to neuroblastoma and Wilms tumor,
genetic and molecular study of hepatoblastoma isretarded. It is necessary to examine genes related togenesis and progression of hepatoblastoma. To examine many kind of genes, gene chip is very effective
method, and we are collaborating with ResearchCenter for Advanced Science and Technology of
University of Tokyo. Using this method and compar-

213

ing gene expression profiles of hepatoblastoma, normal liver, and cirrhotic liver, we have found some
genes that specifically expressed in hepatoblastoma.

Future Prospects
One very important aspect of study in our department is establishment of new area of medicine “fetology”. To know and to treat patients with various congenital anomalies, it is mandatory to know fetal
development of normal baby and babies with congenital anomalies. Development of imaging methods inthese ten years is enormous. However, we need not
only information of structures of fetuses, but alsofunctions of various organs during fetal life. With thedevelopment of new technology which will make itpossible, we can know the physiology of fetus and theeffects of fetal conditions to the health after birth inchildren and adults.
In this 20 years, prognosis of pediatric malignancies in general has improved dramatically. However
there are small fraction of patients whose tumors arevery resistant to standard treatment, and are apt torecur after a short period of remission. These tumors
probably has some genetic characteristics, and if we
can find out some of these characteristics, we can
improve the method of therapy and also can improve
the prognosis of these patients.

Select Publications
1. Yoshida Y, Kobayashi E, Nozawa M, Uchida H,
Fujimura A, Yamanaka T, Miyata M, Hepatic lymphocyte transplantation in hyperbilirubinemic gunnrats. Eur Surg Res 32 (4) : 223-227, 2000
2. Kitano Y, Ruchelli E, Weiner S, Adzick N S. Hepatic
Mesenchymal Hamartoma associated with mesenchymal stem villous hyperplasia of the placenta.
Fetal Diagn Therap 15 : 134-138, 2000
3. Oida T, Suzuki K, Nanno M, Kanamori Y, Saito H,
Kubota E, Kato S, Itoh M, Kaminogawa S, Ishikawa
H. Role of gut cryptopatches in early extrathymic
maturation of intestinal intraepithelial T cells. J
Immunol 164 : 3616-3626, 2000
4. Kitano Y, Flake AW, Quinn T M, Kanai M, Davies P,
Sablich T J, Schneider C, Adzick N S, von Allmen
DLung growth induced by tracheal occlusion in the
sheep is augmented by airway pressurization. J
Pediatr Surg 35 : 216-222, 2000
5. Komuro H, Makino S, Tahara K. Choledochal cyst
associated with duodenal obstruction. J Pediatr Surg
35(8) : 1259-62, 2000
6. Milner R, Kitano Y, Olutoye O, Flake A W, Adzick N
S. Radiofrequency thermal ablation (RTA): a potential treatment for hydropic fetuses with a large chest
mass. J Pediatr Surg 35 : 386-389, 2000
7. A K, Kitano Y, Kriss A, Hensley H, Bergey P, Flake
A, Hubbard A, Leigh Jr J S. In vivo, in utero micro-

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scopic magnetic resonance imaging: application in a
rat model of diaphragmatic hernia. Magn Reson Med
44 : 331-335, 2000
8. Islam A, Kageyama H, Hashizume K, Kaneko Y,
Nakagawara A. Role of Survivin, whose gene ismapped to 17q25, in human and identification of anovel dominant-negative isoform, Survivin β/2B .
Med Pediatr Oncol 35 : 550-553, 2000
9. Kanamori Y, Suzuki K, Oida T, Ishikawa H. Gut-associated lymphoid tissue “Cryptopatches” and intraintestinal development of murine T cells. Mucosal
Immunology Update 3-5, 2000
10. Islam A, Kageyama H, Takada N, Kawamoto T,
Takayasu H, Isogai E,Ohira M, Hashizume K, Kaneko
Y, Nakagawara A. High expression of Survivin,mapped to 17q25, is significantly associated withpoor prognostic factors and promotes cell survival in
human neuroblastoma. Oncogene 19 : 617-623, 2000
11. Ohira M, Kageyama H, Mihara M, Furuda S, Machida
Y, Takayasu H, Islam A, Nakamura Y, Takahashi M,
Shishikra T, Kaneko Y, Toyota A, Sattori S, Sakaki Y,
Ohki M, Horri A, Hayashi Y, Soeda E, Inazawa J, Seki
N, Kuma H, Nozawa I, Nakagawara A. Identificationand characrerization of a 500-kb homozygously deleted region at chromosome 1p36.2 in a human neuroblastoma cell line. Oncogene 19 : 4302-4307, 2000
12. Emura T, Asashima M, Hashizume K. An
Experimental Animal Model of Split Cord
Malformation. Pediatr Neurosurg 33ÅF283-292, 2000
13. Komuro H, Makino S, Tahara K. Laparoscopic resection of an adrenal neuroblastoma detected by mass
screening that grew in size during the observation
period. Surg Endosc 14(3) : 297, 2000
14. Kawarasaki H, Hashizume K, Makuuchi M. Live
Related Pediatric Liver Transplantation. In DK Gupta(Ed.). Textbook of Neonatal Surgery, New Delhi,
Modern Publishers, 202-206, 2000
15. Uchida H, Tanaka H, Kitoh Y, Yanagisawa K, To H,
Tago K, Mizuta K,Fujimura A, Tominaga S,
Hashizume K, Kobayashi E. Mechanisms of
immunomodulation induced by bloodtransfusion:
identification of humoral factors. Transplant Proc 32(2) : 255-256, 2000
16. Ohmori M, Miyashita F, Uchida H, KitohY,Tsuruoka
S, Harada K, Sugimoto K, Fujimura A, Kobayashi E.
Effect of erythromycin on ischemia-reperfusion
injury of liver in rats. Transplant Proc 32 (4) : 811814, 2000
17. Harihara Y, Makuuchi M, Kawasaki S, Hashikura Y,
Kawarasaki H, Takayama T, Kubota K, Ito M, Mizuta
K, Yoshino H, Hirata M, Kita Y, Sano K, Hisatomi S,
Kusaka K, Hashizume K. Influence of HLA compatibility on living-related liver transplantation.
Transplant Proc 32 : 2107, 2000
18. Harihara Y, Makuuchi M, Kawarasaki H, Takayama
T, Kubota K, Ito M,Mizuta K, Yoshino H, Hirata M,
Kita Y, Sano K, Hisatomi S, Kusaka K,Miura Y, Taniai
N, Asato H, Nakatsuka T, Hashizume K. Living-related liver transplantation in adults compared with children. Transplant Proc 32 : 2160-2161, 2000

19. Harihara Y, Sano K, Makuuchi M, Kawarasaki H,
Takayama T, Kubota K, Ito M, Mizuta K, Yoshino H,
Hirata M, Kita Y, Hisatomi S, K, Miura Y, Hashizume
K. Influence of donor age on the graft function after
living-related liver transplantation. Transplant Proc
32 : 2164-2165, 2000
20. Harihara Y, Sano K, Makuuchi M, Kawarasaki H,
Takayama T, Kubota K, Ito M, Mizuta K, Yoshino H,
Hirata M, Kita Y, Hisatomi S, Kusaka K, Miura Y,
Hashizume K. Correlation between graft size and necessary tacrolimus dose after living-related liver transplantation. Transplant Proc 32 : 2166-2167, 2000
21. Hirata M, Harihara Y, Kita Y, Hisatomi S, Miura Y,
Yoshino H, Mizuta K, Ito M, Sano K, Kusaka K,
Kawarasaki H, Kubota K, Takayama T, Hashizume K,
Makuuchi M. Impact of HLA matching in livingrelated liver transplantation..Transplant Proc 32 :
2108-2109, 2000
22. Hirata M, Harihara Y, Hisatomi S, Miura Y, Yoshino
H, Mizuta K, Ito M, Sano K, Taniai N, Kusaka K, Kita
Y, Kawarasaki H, Kubota K, Takayama T, Hashizume
K, Makuuchi M. Living-related liver transplantation
for patients with primary biliary cirrhosis.
Transplant Proc 32 : 2208-2209, 2000
23. Hirata M, Harihara Y, Hisatomi S, Miura Y, Yoshino
H, Mizuta K, Ito M, Sano K, Taniai N, Kusaka K, Kita
Y, Kawarasaki H, Kubota K, Takayama T, Makuuchi
M. A case of esophageal variceal rupture following
acute portal vein thrombosis three days after livingrelated liver transplantation. Transplant Proc 32 :
2266-2268, 2000
24. Tahara K, Uchida H, Yasunaru S, Hakamata Y,
Kobayashi E. Chimeric patterns after organ transplants using green fluorescent protein-transgenic
rat.. American J transplant 1: 336, 2001
25. Kawase E, Tanaka K, Honna T, Azuma N. A case of
atypical WAGR syndrome with anterior segment
anomaly and microphthalmos. Arch Ophthalmol
119: 1855-1856, 2001
26. Utsuki T, Hashizume K, Iwamori M. Impaired spreading of surfactant phospholipids in the lungs of newborn rats with pulmonary hypoplasia as a model ofcongenital diaphragmatic hernia induced by nitrofen
27. Hakamata Y, Tahara K, Uchida H, Sakuma Y,
Nakamura M, Kume A, Murakami T,Takahashi M,
Takahashi R, Hirabayashi M, Ueda M, Miyoshi
I,Kasai N, Kobayashi E. Green fluorescent proteintransgenic rat: a tool for organ transplantation
research..Biochem Biophys Res Commun 286 (4) :779-85, 2001
28. Takayasu H, Horie H, Hiyama E, Matsunaga T,
Hayashi Y, Watanabe Y, Suita S, Kaneko M, Sasaki F,
Hashizume K, Ozaki T, Furuuchi K, Tada M,
Ohnuma N, Nakagawara Al Frequent deletions andmutations of the beta-catenin gene are associatedwith overexpression of cyclin D1 and fibronectin
and poorly differentiated histology in childhood
hepatoblastoma. Clin Cancer Res 7(4): 901-8, 2001
29. Kanamori Y, Hashizume K, Sugiyama M, Morotomi
M, Yuki N. Combination therapy with

Bifidobacterium breve, Lactobacillus casei, and galactooligosaccharides dramatically improved the intestinal function in a girl with short bowel syndrome A
novel synbiotics therapy for intestinal failure. Dig
Dis Sci 46: 2010-2016, 2001
30. Sugawara Y, Makuuchi M, Takayama T, Mizuta K,
Kawarasaki H, Imamura H, Hashizume K. Livertransplantation using a right lateral sector graft froma living donor to her granddaughter.
Hepatogastroenterology 48: 261-63, 2001
31. Sugawara Y, Makuuchi M, Takayama T, Imamura H,
Dowaki S, Mizuta K, Kawarasaki H, Hashizume K.
Small-for-size grafts in living-related liver transplantation. J Am Coll Surg 192: 510-13, 2001
32. Tahara K, Uchida H, Kawarasaki H, Hashizume K,
Kobayashi E. Experimental small bowel transplantation using newborn intestine in rats: III. Long-term
cryopreservation of rat newborn intestine. J Pediatr
Surg 36 (4) : 602-4, 2001
33. Uchida H, Tahara K, Takizawa T, Inose K, Yashiro T,
Hashizume K, Ikeda H, Takahashi M, Kobayashi E.
Experimental small bowel transplantation using
newborn intestine in rats: IV. Effect of cold preservation on graft neovascularization. J Pediatr Surg 36(12) : 1805-10, 2001
34. Sugawara Y, Mizuta K, Kawarasaki H, Takayama T,
Imamura H, Makuuchi M. Risk factors for acuterejection in pediatric living related liver transplantation: The impact of HLA matching. Liver Transpl 7:
769-73, 2001
35. Sugawara Y, Makuuchi M, Takayama T, Yoshino H,
Mizuta K, Kawarasaki H. Liver transplantation fromsitus inversus to situs inversus. Liver Transpl 7: 82930, 2001
36. Mizuta K, Kita Y, Harihara Y, Takayama T, Kubota K,
Hirata M, Inoue K, Sugawara Y, Sakamoto Y, Ito M,
Tanaka H, Inoue Y, Oka T, Kawarasaki H,
Hashizume K, Kawasaki S, Makuuchi M. Colectomy
for uncontrolled hemorrhagic cecal ulcers due to primary cytomegalovirus infection in living-related livertransplantation. Medicine of the Americas 2: 135138, 2001
37. Nakao A, Ogino Y, Tahara K, Uchida H, Kobayashi E
Orthotopic intestinal transplantation using the cuff
method in rats: a histopathological evaluation of theanastomosis. Microsurgery. 21 (1) : 12-5, 2001
38. Sakamoto J, Takata A, Fukuzawa R, Kikuchi H,
Sugiyama M, Kanamori Y, Hashizume K, Hata J. A
Novel WT1 Gene Mutation Associated with Wilms’
Tumor and Congenital Male Genitourinary
Malformation. Pediatric Research 50 : 337-344, 2001
39. Komuro H, Makino S, Momoya T, Uehara Y, Tahara
K, Momoi M: Cholangitis associated with cystic
dilatation of the intrahepatic bile ducts after antireflux valve construction in biliary atresia. Pediatr Surg
Int 17 (2-3) : 108-10, 2001
40. Hirata M, Harihara Y, Kitamura T, Hisatomi S, Kato
M, Dowaki S, Mizuta K, Sugawara Y, Kita Y, Kubota
K, Takayama T, Kawarasaki H,Hashizume K,
Makuuchi M: The influence of donor age to graft vol-215

ume increase rate in living donor Liver transplantation. Transplant Proc 33: 1416-17, 2001
41. Kanamori Y, Hashizume K, Kawarasaki H, Kitano Y,
Sugiyama M, Tanaka Y: Single vaginal ectopic ureterand renal hypoplasia associated with urogenital
sinus and abdominal muscular hypoplasia-a novel
subtype of prune-belly syndrome in a female child?
Urology 57: 975vi-ix, 2001
42. Komuro H, Makino S, Yasuda Y, Ishibashi T, Tahara
K, Nagai H. Pancreatic complications in choledochal
cyst and their surgical outcomes. World J Surg 25(12) : 1519-23, 2001
43. Tatsumi K, Taki T, Taniwaki M, Nakamura H,
Taguchi J, Chen YZ, Bessho F, Yanagisawa M,
Hayashi Y: The CDCREL1 gene fused to MLL in de
novo
acute
myeloidleukemiawitht(11;22)(q23;q11.2) and its frequent expression in
myeloid leukemia cell lines. Genes Chromosomes
Cancer 30:230-235, 2001
44. Murata T, Kurokawa R, Krones A, Tatsumi K, Ishii
M, Taki T, Masuno M, Ohashi H, Yanagisawa M,
Rosenfeld MG, Glass CK, Hayashi Y: Defect of histone acetyltransferase activity of the nuclear transcriptional coactivator CBP may cause RubinsteinTaybi syndrome. Hum Mol Genet 10: 1071-1076,
2001
45. Jamal R, Taketani T, Taki T, Bessho F, Hongo T,
Hamaguchi H, Horiike S, Taniwaki M, Hanada R,
Nakamura H, Hayashi Y: Co-duplication of the MLL
and FLT3 gene in patients with acute myeloid
leukemia. Genes Chromosomes Cancer 31: 187-190,
2001
46. Niitsu N, Hayashi Y, Sugita K, Honma Y:
Sensitization by 5-aza-2’-deoxycytidine of leukaemia
cells with MLL abnormalities to induction of differentiation by all-trans retinoic acid and 1α, 25-dihydroxyvitamin D3. Br J Haematol 112:315-326, 2001
47. Niitsu N, Hayashi Y, Honma Y: Downregulation of
MLL-CBP fusion gene expression is associated withdifferentiation of SN-1 cells with t(11;16)(q23;p13).
Oncogene 20: 375-384, 2001
48. Ohnishi H, Hayashi Y, Okamura T, Tabuchi K,
Kobayashi M, Bessho F: An infant witht(4;11)(q21;q23) acute lymphoblastic leukemia successfully treated using immunomagnetically purged
autologous bone marrow transplantation. Med
Pediatr Oncol 36: 388-389, 2001
49. Shibuya N, Taki T, Mugishima H, Chin M, Tsuchida
M, Sako M, Kawa K, Ishii E, Miura I, Yanagisawa M,
Hayashi Y: t(10;11)-acute leukemias with MLL-AF10
and MLL-ABI1 chimeric transcripts; specific expression patterns of ABI1 gene in leukemia and solid
tumor cell lines. Genes Chromosomes Cancer 32:110, 2001.
50. Liu M, Taketani T, Li R, Takita J, Taki T, Yang HW,
Kawaguchi H, Ida K, Matsuo Y, Hayashi Y: Loss of
p73 gene expression in lymphoid leukemia cell lines
is associated with hypermethylation. Leuk Res
25:441-447, 2001

216

Department of Geriatric Medicine /Department of Aging Research
Outline and Research Objectives
The Department of Geriatrics was established in1962, as the first geriatric department inJapan.
Since elderly patients usually have multiple organ disorders, we have to take care of thepatients as whole from multiple points of view. In addition, in the elderly patients, symptoms and signs, responses to the treatment are sometimes quite different from the young.
We have to have a broad knowledge on the physiological and metabolic changes with aging
when we treat the elderly patients. Quality of life of the patients is another point of viewwhich should be emphasized.
Our sub-specialty includes respirology, cardiology, neurology, hematology, bone metabolism, besides the general geriatric internal medicine. We are trying to elucidate the pathophysiology of aging process and elderly patients from viewpoints of basic aging science
using molecular biology technique and clinical aspects using the recent advancement oftechnology and geriatric assessment.
There are 19 plus additional beds about 6 in the clinical ward, which are under thesupervision of the residents and assistant staff members. An assistant staff member withmore than 10 years experience teams up with the resident on a man-to-man basis, giving
instruction as to actual clinical decision making and procedures. The final decision making
is performed during the weekly Professor’s rounds.
We have also developed new geriatric scale such as vitality index. This is a unique index
for assessment of energy and vitality in frail elderly patients. Comprehensive geriatricassessment and critical pass applied for the treatment and care of the patients. Based onthese comprehensive approaches, both staffs of medical social service and welfare divisionof Tokyo University Hospital and we develop the new system which bridge between inpatients and out-patients in terms of maintaining a constant level of quality of life ofpatients and caregivers.
Specialized services are provided to out-patients on a daily in all areas internal medicine.
Approximately 300 new and a total of 16,000 patients visit the out-patients clinic in a year.

Faculties and Students
Professor and Chair
Lecturer

Yasuyoshi Ouchi,M.D. (1995~)
Yoshio Namba,M.D.
Takahide Nagase,M.D.
Satoshi Inoue,M.D.
Koichi Kozaki, M.D.
Associate .................................6
Postdoctoral Fellow ................4
Graduate Student....................8
Research Student ....................5
Secretary .................................6

Past Research and Major Accomplishments
Lung diseases including pneumonia are major
causes of death in the elderly. In respiratory diseases,
there are several inflammatory disorders to which no
pharmaceutical agents are currently effective. For

example, adult respiratory distress syndrome (ARDS)
is an acute lung injury and the mortality rate for
ARDS ranges from 40-70% despite of intensive careusing currently available drugs. Idiopathic pulmonary
fibrosis (IPF) is a progressive and fatal disease, whilecurrent medical intervention for IPF is only oxygentherapy except for lung transplantation. However,their mechanisms still remain to be elucidated.
Therefore, we have aimed to elucidate the pathophysiological and molecular mechanisms underlying
inflammatory diseases. Our recent studies haveshown that various mediators including cytokines,
eicosanoids or adhesion molecules are involved in thedevelopment of inflammatory lung disorders.
Especially, platelet-activating factor (PAF) and metabolites of arachidonic acid, i.e., eicosanoids, are lipid
mediators that have various biological effects. PAF is aproinflammatory phospholipid mediator that exerts
its action via activation of G-protein-coupled PAF

217

receptor (PAFR). A key enzyme for the production ofthese inflammatory mediators including eicosanoids
and PAF is cytosolic phospholipase A2 (cPLA2). To
perform this study, we used mutant mice, i.e., PAFR
transgenic mice, PAFR gene-disrupted mice, andcPLA2 gene-disrupted mice, in collaboration with
Department of Biochemistry. Our observations suggest that both PAF and cPLA2 are involved in thepathogenesis of acute lung injury and pulmonary
fibrosis. The inhibition of these pathways might provide a novel therapeutic approach to ARDS and IPF.
We have also examined the host-defense system,
especially antimicrobial peptides including defensins.
Recently, we have discovered a novel mouse betadefensin, mBD-6, predominantly expressed in skeletal
muscle. In humans and mice, we have also identified
multiple novel epididymis-specific beta-defensin isoforms. These finding suggest novel physiological rolesof this peptide family.
Cardiovascular research laboratory has been performing research of atherosclerosis. It is generally
accepted that premenopausal women are protectedfrom atherosclerotic diseases because female hormone, especially estrogen, appears to have a beneficial
effect on vascular wall. We have identified that vascular smooth muscle cells (VSMC) expressestrogen
receptor (ER) and these cells respond to estrogen fromthe evidence that estrogen inhibits endothelin-1 production and c-fos gene expression in rat aorta andestrogen inhibits migration and proliferation of vascular smooth muscle cells. We also found estrogenexerts beneficial effect in vivo by showing that estrogen inhibits intimal thickening of rat femoral arteryinduced by cuff-placement around the artery. With
respect to endothelial cells (EC), another major component constituting the vascular wall, we found that
estrogen attenuates endothelin-1 production inbovine endothelial cells via estrogen receptor and that
estrogen prevents oxidative stress-induced endothelial cell apoptosis in rats.
Besides estrogen, we have revealed that red wine
polyphenols (RWPs) have inhibitory roles in VSMC
function. RWPs inhibit VSMC proliferation, which ismediated by cyclin A downregulation. RWPs also
inhibit VSMC migration via two distinct signaling
pathways, phosphatidylinositol-3’ kinase and p38
mitogen-activated protein kinase. We also showed
potential involvement of various vasoactive substances, calcitonin gene-related peptide, parathyroid
hormone-related protein, vasopressin, and activin-A,
in the pathogenesis of atherosclerosis.
From the clinical perspective, our laboratory has auseful technique to evaluate vascular function, flowmediated vasodilatation (FMD). With this approach,we have shown that endothelium-dependent vasodilatation of the brachial artery is increased in the follic-218

ular and luteal phase of women’s menstrual cycle
when serum estradiol level is high, indicating that
endogenous estradiol is involved in menstrual cyclerelated vasodilatation. We also found that FMD is
impaired in obese men with visceral fat accumulation,
FMD negatively correlates with intima-media thickness in the carotid artery in men, long-term reduceddose hormone replacement therapy improves FMD
and intima-media thickness in the carotid artery inpostmenopausal women.
Currently, most of the research activities in neurological laboratory focus on the molecular aspects of
Alzheimer’s disease(AD). The main research topics are
elucidating the molecular mechanism underlying theaccumulation of amyloid b-protein, determining theimplications of apolipoprotein E (apoE ε4) genotype
and various risk factors in AD.
Endocrine/Metabolism Research Laboratory focuses on the clinical and basic research concerning
endocrine and metabolic diseases. We are particularlyinterested in the pathophysiological mechanisms inosteoporosis and hormone-dependent cancers.
Regarding osteoporosis, we have shown that genetic
factors are very important to develop the disease byanalyzing polymorphisms of the genes relating bonemetabolism. On the other hand, we recently haverevealed that an estrogen responsive RING finger protein Efp takes a critical role in estrogen-dependent
growth control of breast tumor, by proteolysis of acell-cycle checkpoint 14-3-3 sigma (Nature 2002, 417,
871). This finding may provide a new tool for diagnosis and treatment of breast tumor, especially of tumor
that is resistant to anti-estrogenic drugs.
We have also studied disuse syndrome as well as
the program of prevention and treatment of that.
Now we are investigating many aspects of elderly
patients by using newly developed geriatric scale.

Current Research
To further investigate the pathophysiological andmolecular mechanisms underlying inflammatory diseases, we continue to study the roles of various mediators including cytokines, eicosanoids, adhesion molecules, and CGRP family in the development of inflammatory lung disorders. Especially, CGRP family peptides including CGRP and adrenomedullin are neuropeptides that have various biological actions such as
responses to sensory stimuli, cardiovascular regulation and vasodilation. Based on their physiologicalroles, it is assumed that CGRP family might beinvolved in the pathogenesis of inflammatory diseases
including bronchial asthma. Currently, in collaboration with Department of Cardiovascular Medicine, weare studying the pathophysiological role of CGRP family using mutant mice deficient in either CGRP oradrenomedullin.
High incidence of respiratory infection including
pneumonia is a major characteristic of geriatric disease, while little is known about its mechanism. We
currently hypothesize that ageing process might affectthe expression pattern of defensins, leading to thesusceptibility to infection in the elderly. To address
this question, we are studying the ageing effects on
the expression pattern of defensins using mice. In
addition, to better understand the pathophysiologicalroles of defensins, we are now developing the mutant
mice deficient in defensins.
We are currently pursuing the differential role ofestrogen receptors, ERα and β, in VSMC function
and in other cell types utilizing adenoviral overexpression system, because we have previously shown that
estrogen has anti-proliferative and anti-migratory
effects in VSMC (Atherosclerosis. 1997). We are also
trying to overexpress each ER subtype in the aorta oflaboratory animals to identify the differential role of
ER in estrogen’s action. We are studying the role of
selective estrogen receptor modulator such as raloxifene to explore the usefulness of this new estrogenmodifying drug.
Our laboratory has been investigating vascular calcification, one of the hallmarks of vascular aging. We
have thus far found that VSMC calcify when cultured
in the presence of high inorganic phosphate, the phenomenon augmented by the addition of vitamin D3,
dexamethasone and advanced glycation end products,
attenuated by bisphosphonate and HMG CoA reductase inhibitor. We are currently studying the mechanism how VSMC calcification is regulated.
It is well known that flow-mediated vasodilatation(FMD) is under much influence by nitric oxide (NO).
Because it has been shown that an endogenous
inhibitor of NO synthase, asymmetric dimetylarginine(ADMA), and its hydrolyzing enzyme, NG,NGdimethylarginine dimethylaminohydrolase (DDAH),
are circulating in human blood, we are interested in
the role of ADMA and DDAH in such pathophysiologic conditions as pre- and post-female hormone
replacement therapy and pre- and post-treatment of
sleep apnea syndrome using nasal continuous positive
airway pressure.
The ε4 allele of apoE is a major risk factor for both
sporadic and late-onset familial AD. The apoE receptor family consists of cell-surface receptors that recognize extracellular ligands and internalize them for
degradation by lysosomes. In order to gain insightabout these receptors in the CNS, we raised a rabbit
polyclonal antibody and examined immunohistochemically human brain tissue. The immunoreactivity
was found in senile plaque in AD. These results may
suggest a role of apoE receptor in amyloid formation.
As estrogen is one of the key factors that candetermine the stages and prognosis of osteoporosis
and hormone-dependent cancers, we are particularly
studying the molecular mechanism of estrogen in its
target organs. Biochemical and genetic approaches are
utilized for the project, including analysis using
osteoblasts and tumor cell lines, characterization ofanimal models targeting estrogen receptors and estrogen-responsive genes, genetic analysis using clinicalsamples from patients.

Future Prospects
Major causes of death in the elderly include respiratory diseases including pneumonia, especially aspiration pneumonia. Furthermore, there are fatal
inflammatory disorders such as ARDS and IPF, towhich no useful drugs are currently available. We
expect that the inhibition of PAF and cPLA2 pathways might provide a novel therapeutic approach to
ARDS and IPF, leading to the development of a revolutionary medicine.
Genetic features including single nucleotide polymorphism (SNP) are potentially associated with the
etiology of asthma. The current study may suggestthat CGRP family genes could be targets for SNP
research. High incidence of respiratory infection
including pneumonia in the elderly might be
explained by the mechanism that ageing process
might alter the expression pattern of defensins, leading to the immunosupression and the susceptibility toinfection. We expect that our ongoing study related todefensins could make a contribution to the innateimmunity researches. Furthermore, novel roles ofdefensins might potentially provide a novel therapeutic approach to infectious diseases.
Because the aging process and age-related diseases
are closely associated with vascular aging, to prevent
atherosclerosis is a way to improve physical activity inthe elderly. Our laboratory keeps pursuing the mechanism of atherosclerosis, especially how hormone andits receptor regulate vascular function. We will keep
performing research as to how sex hormones or theirrelated biological materials modulate vascular function in vitro and in vivo. We will also try to elucidate
the mechanism how we can reduce vascular calcification, because vascular calcification appears tightly
linked with vascular aging, which is one of our interests.
We are studying the genetic and environmental
risk factors and prevalence, and incidence of
Alzheimer’s disease among the elderly in Japan.
Because estrogen is one of the key factors that candetermine the stages and prognosis of osteoporosis,
hormone-dependent cancers, and age-related changesof females, we are pursuing prevention and new treatment of osteoporosis and cancer. It may be contribute219

to maintain the constant level of physical and social
activity in elderly peoples. Furthermore, our basic
research focuses on novel findings of new moleculartargets for drug development concerning osteoporosis
and hormone-dependent cancers.
Based on these clinical and basic science data, weare pursuing prevention of a variety of age-related diseases and geriatric syndrome. We also propose standards for facilitation of care across the health service
continuum, care of the nursing home resident, andpalliative and hospice care.

Research Grants
Yasuyoshi Ouchi
1. Grant-in Aid (No.13557062) for Scientific Researchfrom the Minstry of Education, Science and Cultureof Japan. Elucidation of the differential role of estrogen receptor a and b in vascular lesion formation.
1999-2001 (three year research foundation grant B2).
2. Funds for Comprehensive Research on Aging and
Health, Japan Foundation for Aging and Health.
Hormone replacement therapy for elderly women.
Yasuyoshi Ouchi 1998-2000 (three year grant, H10tyojyu-007).
Nagase Takahide
1. Grant-in Aid(No. 12470134) for Scientific Researchfrom the Ministry of Education, Science, Sports andCulture of Japan. The study of molecular mechanisms underlying ARDS using genetically-engineered
mice to develop a novel treatment (three year
research foundation grant B).
2. Health Science Research Grant-in Aid forComprehensive Research on Aging and Health from
the Ministry of Health, Labour and Welfare of Japan.
The study of pathophysiological mechanisms underlying inflammatory/intractable lung diseases in theelderly: a strategic development of novel therapeutic
approaches (H14-).
Yoshio Namba
1. Health Science Research Grant-in Aid for Brain
Science from the Ministry of Health and Welfare ofJapan. Genetic and environmental risk factors for
Alzheimer’s disease (three year grant, H10-H12).

Select Publications
1. Urano T, Saito T, Tsukui T, Fujita M, Hosoi T,
Muramatsu M, Inoue S: Efp targets 14-3-3sigma forproteolysis and promotes breast tumour growth.
Nature 417, 871-875, 2002.
2. Nagase T, Uozumi N, Ishii S, Kita Y, Yamamoto H,
Ohga E, Ouchi Y, Shimizu T. A pivotal role of cytosolic phospholipase A2 in bleomycin-induced pulmonary fibrosis. Nature Med 8: 480-484, 2002.

220

3. Nagase T, Ishii S, Shindou H, Ouchi Y, Shimizu T.
Airway hyperresponsiveness in transgenic mice overexpressing platelet activating factor receptor is mediated by an atropine sensitive pathway. Am J Respir
Crit Care Med 165: 200-205, 2002.
4. Hashimoto M, Miyao M, Akishita M, Hosoi T, Toba
K, Kozaki K, YoshizumiM, Ouchi Y. Effects of longterm and reduced-dose hormone replacement therapy on endothelial function and intima-media thickness in postmenopausal women. Menopause. 9:5864, 2002.
5. Iijima K, Yoshizumi M, Hashimoto M, Akishita M,
Kozaki K, Ako J, Watanabe T, Ohike Y, Son B, Yu J,
Nakahara K, Ouchi Y. Red wine polyphenols inhibit
vascular smooth muscle cell migration through two
distinct signaling pathways. Circulation. 105:24042410, 2002.
6. Kageyama A, Hirano T, Kageyama H, Osaka T,
Namba Y, Tsuji M, Adachi M, Inoue S. Distinct roleof adiposity and insulin resistance in glucose intolerance: studies in ventromedial hypothalamic-lesioned
obese rats. Metabolism. 51:716-23, 2002.
7. Kaneki M, Hedges SJ, Hosoi T, Fujiwara S, Lyons A,
Crean SJ, Ishida N, Nakagawa M, Takechi M, Sano Y,
Mizuno Y, Hoshino S, Miyao M, Inoue S, Horiki K,
Shiraki M, Ouchi Y, Orimo H: Japanese fermented
soybean food as the major determinant of the large
geographic difference in circulating levels of vitamin
K2: possible implications for hip-fracture risk.
Nutrition 17, 315-21, 2001.
8. Sudoh N, Toba K, Akishita M, Ako J, Hashimoto M,
Iijima K, Kim S, Liang YQ, Ohike Y, Watanabe T,
Yamazaki I, Yoshizumi M, Eto M, Ouchi Y. Estrogen
prevents oxidative stress-induced endothelial cell
apoptosis in rats. Circulation. 103:724-729, 2001.
9. Yamaguchi Y, Fukuhara S, Nagase T, Tomita T,
Hitomi S, Kimura S, Kurihara H, Ouchi Y. A novel
mouse beta-defensin, mBD-6, predominantly
expressed in skeletal muscle. J Biol Chem 276:
31510-31514, 2001.
10. Ishikawa M, Akishita M, Kozaki K, Toba K, Namiki
A, Yamaguchi T, Orimo H, Ouchi Y. Expression ofparathyroid hormone-related protein in human andexperimental atherosclerotic lesions: functional rolein arterial intimal thickening. Atherosclerosis.
152:97-105, 2000.
11. Inoue S, Ogawa S, Horie K, Hoshino S, Goto W,
Hosoi T, Tsutsumi O, Muramatsu M, Ouchi Y: An
estrogen receptor beta (ER beta) Isoform that lacks
exon 5 has dominant negative activity on both ER
alpha and ER beta. Biochem Biophys Res Commun
279, 814-819, 2000.
12. Miyao M, Hosoi T, Emi M, Nakajima T, Inoue S,
Hoshino S, Shiraki M, Orimo H, Ouchi Y:
Association of bone mineral density with a dinucleotide repeat polymorphism at the calsitonin (CT)locus. J Hum Genet 45: 346-350, 2000.
13. Ogawa S, Emi M, Shiraki M, Hosoi T, Ouchi Y, Inoue
S: Association of estrogen receptor beta (ESR2) gene

polymorphism with blood pressure. J Hum Genet 45:
327-330, 2000.
14. Ogawa S, Fujita M, Ishii Y, Tsurukami H,
Hirabayashi M, Ikeda K, Orimo A, Hosoi T, Ueda S,
Nakamura T, Ouchi Y, Muramatsu M, Inoue S:
Impaired estrogen sensitivity in bone by inhibiting
both estrogen receptor alpha and beta pathways. J
Biol Chem 275: 21372-21379, 2000.
15. Hoshino S, Hosoi T, Miyao M, Shiraki M, Orimo H,
Ouchi Y, Inoue S: Identification of a novel polymorphism of estrogen receptor alpha gene that is associated with calcium excretion in urine. J Bone Miner
Metab 18: 153-157, 2000.
16. Urano T, Hosoi T, Shiraki M, Toyoshima H, Ouchi Y,
Inoue S: Possible involvement of p57Kip2 gene in thebone metabolism. Biochem Biophys Res Commun
269: 422-426, 2000.
17. Miyao M, Morita H, Hosoi T, Kurihara H, Inoue S,
Hoshino S, Shiraki M, Yazaki Y, Ouchi Y: Associationof methylenetetrahydrofolate reductase (MTHFR)
polymorphism with bone mineral density in postmenopausal Japanese women. Calcif Tissue Int 66:
190-194, 2000.
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Department of
Cardiothoracic Surgery
Outline and Research Objectives
Our department specializes in all areas of cardiothoracic surgery including congenital andacquired cardiac diseases, thoracic and abdominal aortic diseases, and benign and malignant
diseases of respiratory and mediastinal organs, except for diseases of the esophagus andmammary glands. The Department of Cardiothoracic Surgery, University of Tokyo, wasestablished in December 15, 1964 as the first department of this field in the Japanesenational universities. Since then it has played an internationally leading role and contributed to development of the field. Professors and Chairs in the history of the department
are as follows; Seiji Kimoto (1964.12.15 ~ 1968.3.31), Masahiro Saigusa (1968.4.1 ~1981.3.31), Ken-ichi Asano (1981.4.1 ~ 1986.3.31), Akira Furuse (1986.4.1 ~ 1997.3.31) and
Shinichi Takamoto (1997.6.1 ~). The cardiothoracic department of the University of Tokyo
has created highly active research programs in the every field of cardiothoracic surgery.
Recently, most of the basic research activities are focused in the fields of brain and spinal
cord protection during aortic surgery, homo- and xeno-transplantation of the heart, lung,heart valves and trachea, and surgical oncology of the pulmonary and thymic neoplasms.
Although, there are many other active research projects such as stent-graft repair for thoracic aortic aneurysm, minimally invasive cardiac surgery, and intraoperative real-time 3-D
echocardiography.

Faculties and Students
Professor and Chair Shinichi Takamoto,M.D. (1997-)
Associate Professors Yutaka Kotsuka, M.D
Jun Nakajima, M.D.
Lecturer
Arata Murakami, M.D. andToshiya Ohtsuka, M.D.
Associate .................................8
Graduate Students ................10
Research Students...................1

Past Research and Major Accomplishments
Dr. Takamoto invented a new method of brain
protection during surgery of a distal arch aneurysm
under deep hypothermia (Fig 1). The new method is akind of retrograde cerebral perfusion, which simplify
the retrograde perfusion system for patients of distal
aortic arch aneurysmon. This new method has been
accepted as a standard procedure of a distal arch
replacement in all over the world. The method has
been applied to many patients, and it was clarified
that the method provides a prolongation of safety limits of circulatory arrest and a beneficial effect in prevention of brain embolism.
Allograft tissue has sevral advantages in that antiinfection, softness, and less compliance mismatching.
Cell viability of the tissue can be maintained for morethan hundred years if properly preserved under theliquid nitrogen. We started basic study on homografts

Fig 1

Fig 2

tissue transplantation including the cardiac valve,aorta and trachea 8 years ago. We founded the
Universty of Tokyo Tissue Bank (UTTB) in 1999,
where human cardiac valves, vessels and tracheas

223

obtained from non-brain dead donors are cryopreserved. Cardiac valves are used for the patients ofinfective endocarditis, congenital heart disease.
Vessels are used for artificial graft infection, livertransplant recipients. The number of donor and usageof these allogaft tissus are increasing every year, andmore and more patients are being saved with these
allograft tissues (Fig 2).
As for clinical activities, surgical cases are remarkably increasing as shown in the table below.
Table 1: Patients who underwent surgery in the department
Cardiovascular
General Thoracic
Total

1997
152
178
330

1998
157
202
359

1999
179
170
349

2000
225
197
422

2001
272
185
457

Current Research
Thirteen staff members including professors andlecturers all participate not only in surgeries but alsopatient care on both in-patient and out-patient bases.
They are all engaged in research activities for clinicaland experimental subjects. As the subjects of cardiothoracic surgical fields are so close one another, we do
not have sectioned research groups within our department. Instead, we make a project team for eachresearch subject with most appropriate staff members
and residents. Every associate takes part in at least
one to several project teams. Main subjects of current
research includes brain and spine protection during
aortic surgery, homo- and xeno-transplantation of theheart, lung, heart valves and and trachea, surgicaloncology of the pulmonary and thymic neoplasms,
stent-graft repair for thoracic aortic aneurysm, minimally invasive cardiac surgery, and intraoperative 3-D
echocardiography.
Spinal cord protection: Paraplegia remains one of
the most serious complications after thoracoabdominal aortic aneurysm repair. In order to prevent paraplegia, it is important to identify and preserve criticalsegmental arteries. We developed new method to
identify critical segmental artery. The ultrasonographic evaluation of the hemodynamics of intercostal
arteries was the key to identify critical segmental
artery (Fig 3).
Cryopreserved allotransplantation: Cryopreserved
allogeneic heart valves and vessels have come to be
widely utilized in cardiothoracic surgery because of
their excellent durability. We have studied the allogenicity of the cryopreserved tissues using cell cultures of the airway epithelium, vascular endothelium,and fibroblasts. We have investigated whether cryopreserved tracheal allotransplantation is applicable.
Experimental cryopreserved tracheal allotransplanta-

224

tion was performed in primates, which have a closer
anatomical and immunological relation with humans
than other animals, to confirm the possible clinicalfeasibility of cryopreserved tracheal allotransplantation. Immunogenicity was attenuated by cryopreservation, and cryopreserved tracheal allografts wereincorporated in all animals (Fig 4).
Stent-graft repair: We developed ultrathin-wallvascular grafts with a wall thickness of 42 to 137μm
for endovascular surgery. We studied the physicalproperties of the ultrathin-wall grafts in in vitro
experiments. We conclude that the newly developed
ultrathin-wall grafts are suitable for endovascular surgery (Table 2).
After animal experiments, we applied the grafts toclinical patients with good results. We also developed
auto-swelling vascular grafts, using ultrathin-wall
Dacron vascular grafts and superabsorbent polymer.
These grafts were designed to prevent endoleak following stent-graft repair of aortic aneurysm. Stentgrafts covered with auto-swelling grafts are expected
to have beneficial effects in preventing endoleak.
Minimally invasive cardiac surgery: We have beenusing minimally invasive techniques in various procedures of thoracic and cardiac surgery. We have
employed video-assisted thoracoscopic surgery (VATS)
for pulmonary or mediastinal surgery since 1992. We
have made retrospective studies on feasibility and surgical outcome of VATS for diagnosis and treatment oflung neoplasms. We have concluded that VATS for
diagnosing pulmonary indeterminate nodules and

fig3

fig4

Table 2
Wall thickness (µm)

137

117

95

85

75

64

50

42

Transverse Microscopic
Porosity (µm)

0

0

0

0

0

11.3±13.6

57.4±25.0

75.9±18.8

Longitudinal Microscopic
Porosity (µm)

0

0

0

0

0

36.9±7.8

39.0±7.9

132.3± 7.8

Planimetric Porosity
(µm2)

0

0

0

0

0

416±494

2,238±1193>10,000

Longitudinal Tensile
Strength (Kg)22.5±2.0

19.4±1.2

15.3±0.8

14.1±1.8

13.1±0.9

9.5±0.9

7.4±0.5–

Water Permeability(ml/min/cm2)

330±17220±20250±36180±17380±531,700±361

2,960±443

3,840±668

fig6
fig5

VATS for pulmonary metastasectomy are justified as astandard therapeutics. Off-pump coronary artery
bypass is achieved without hemodynamic assistanceby cardiopulmonary bypass, and it has been applied
to increasing number of patients with multi-vessel
ischemic heart disease. A single bypass from theinternal mammary artery (IMA) to the left anterior
descending coronary artery has been accomplished via
a mini-thoracotomy approach. Valvular disease or atrial septal defect have been repaired via a limited sternotomy using a special retractor and port-accessible
heart-lung machine. Video-endoscopy has been positively used for minimally invasive approach.
Thoracoscopy has been used for mobilization of ITA
graft (Fig 5), and cardioscopy has been used in valvular repair, correction of intra-cardiac anomaly, andleft-atrial cryo-ablation.
Real-time 3-D echocardiography: We have developed real-time three-dimensional echocardiography(RT3DE) and applied it to monitoring heart operations, especially ASD closure, as a means of assistingtreatment (Fig 6). In the animal experiment, ASD was
successfully closed by RT3DE monitoring, and examination of the excised heart showed that all sutures
were located, and the reliability of the images
obtained by RT3DE was confirmed.
Surgical oncology of the thymic neoplasms: Toclassify the thymic epithelial neoplasms, the maturation stage of T-lineage lymphoid cells infiltrating
thymic neoplasms (TIL) was examined by flow-

cytometry to associate it with the degree of tumor
malignancy. CD4+CD8+ or CD10+ T-lineage cells werethe most reliable markers of the benignancy of thymic
epithelial tumors. CD4 / CD8 single positive cells or
CD20-positive cells were characteristic in thymic carcinoma. Flow-cytometry on the maturity of lymphoidcells infiltrating thymic epithelial tumors was feasiblefor determining their degree of malignancy. Recently
we have investigated T-cell receptor (TCR) gene rearrnagement of the TIL. We have observed the appearance of clonal bands in TCR-beta and gamma, suggesting that partial gene rearrangement occurred in the
TIL.

Future Prospects
Cardiac Transplantation is established treatment in
the world. However, this treatment has not been welldeveloped in Japan due to domestic reasons. We are
now preparing clinical cardiac transplantation. We aregoing to develop basic and clinical research on cardiac
transplantation as well as clinical application.

225

Research Grants
1. Grant-in Aid for Scientific Research from theJapanese Ministry of Education, Science, Sports andCulture (A)(1): Studies on management of hearttransplant recipients and maintenance of donororgans. 1998-2000
2. Grant-in-Aid for Publication of Scientific Research
Results: Japan Adult Cardiovascular Surgery
Database. 2002
3. Grant-in-Aid for University and Society
Collaboration: Establishment of Tissue Banking
System on the basis of regional linkage. 2000-2002
4. Grant-in Aid for Scientific Research from theJapanese Ministry of Education, Science, Sports andCulture (B)(2): Basic and clinical study on percutaneous endovascular surgery with ultrathin-wall vascular grafts. 1997-1998
5. Grant-in Aid for Scientific Research from theJapanese Ministry of Education, Science, Sports andCulture (B)(2): Development of auto-swelling vasculargrafts using super-absorbent polymer. 1999-2001

Select Publications
1.

2.

3.

4.

5.

6.

7.

8.

9.

Imanaka, K., Takamoto S. Furuse A. Mitral regurgitation late after Manouguian’s anulus enlargement
and aortic valve replacement. J Thorac Cardiovasc
Surg 115, 727-9, 1998.
Ono, M, Takamoto S, Ohtsuka T. Right ventricularrupture in a minimally invasive direct coronary
artery bypass grafting. Eur Journal Cardiothorac Surg
14, 536-537, 1998.
Imanaka K. Shimizu S. Matsumoto J. Hashizume K.
Tsuchiya K. Takemura T. Unilateral absence of pulmonary artery and ventricular defect in an infant.
Ann Thorac Surg 66, 251-2, 1998.
Kawauchi M, Nakajima J, Takeda M, Oka T,
Takamoto S. Aortic valves are antigenic but less sothan myocardium. J Thorac Cardiovasc Surg 116,
532, 1998.
Miyaji K, Sugiura S, Omata S, Kaneko Y, Ohtsuka T,
Takamoto S. Myocardial tactile stiffness: A variableof regional myocardial function. J Am Coll Cardiol
31, 1165-1173, 1998.
Kubota H, Takamoto S. Takeshita M. Miyaji K.
Kotsuka Y. Furuse A. Atrial ablation with an IRK-151
infrared coagulator. Ann Thorac Surg 66, 95-100,
1998.
Ohtsuka T, Takamoto S, Endoh M, Ono M, Minami
M. Ultrafast computed tomography for minimallyinvasive coronary artery bypass grafting. J Thorac
Cardiovasc Surg 116, 173-4, 1998.
Ohtsuka T, Endoh M, Takamoto S. Minimally invasive left anterior descending coronary artery bypass
with right gastroepiploic artery graft. J Thorac
Cardiovasc Surg 116, 528-9, 1998.
Ohtsuka T. How high should mammary artery be
harvested for the minimally invasive approach? Ann
Thorac Surg 66, 985-6, 1998.

226

10. Kotsuka Y, Tanaka O, Takamoto S, Furuse A.
Intravalvular implantation technique for aortic valve
replacement in aortitis syndrome. Ann Thorac Surg
67, 89-92, 1999.
11. Imanaka K, Takamoto S, Murakami A, Kaneko Y.
Minimally invasive extracardiac conduit replacement
via a left anterior small thoracotomy. J Thorac
Cardiovasc Surg 118, 1124-5, 1999.
12. Imanaka, K, Takamoto S, Kaneko Y, Murakami A,
Shibuya K. Extracardiac conduit replacement withautologous tissue only, via a left anterior small thoracotomy. Thorac Cardiovasc. Surg 47, 251-2, 1999.
13. Imanaka, K, Takamoto S, Murakami A, Kaneko Y.
Right ventricular thrombosis early after bidirectional
Glenn shunt. Ann Thorac Surg 68, 563-5, 1999.
14. Ohtsuka T, Imanaka K, Endoh M, Kohno T,
Nakajima J, Kotsuka Y, Takamoto S. Hemodynamic
effects of carbon dioxide insufflation under singlelung ventilation during thoracoscopy. Ann Thorac
Surg 68, 29-33, 1999.
15. Nakajima J, Poindexter NJ, Hillemeyer PB, Trulock
EP, Cooper JD, Patterson GA, Mohanakumar T,
Sundaresan RS. Cytotoxic T lymphocytes directedagainst donor HLA class I antigens on airway epithelial cells are present in bronchoalveolar lavage fluid
from lung transplant recipients during acute rejection. J Thorac Cardiovasc Surg 117, 565-71, 1999.
16. Nakajima J., Furuse A, Kohno T, Ohtsuka T,
Matsumoto J, Oka T. Transthoracoscopic needle
biopsy for indeterminate lung nodules. SurgicalEndoscopy 13, 386-9, 1999.
17. Tanaka K, Takamoto S, Ohtsuka T, Kotsuka Y.
Advaseal for acute aortic dissection: experimental
study. Eur J Cardio-thorac Surg 15, 114-115, 1999.
18. Tanaka K, Takamoto S, Ohtsuka T, Kotsuka Y,
Kawauchi M. Application of Advaseal for acute aortic
dissection: experimental study. Ann Thorac Surg 68,
1308-1313, 1999.
19. Murakawa T, Nakajima J, Ono M, Murakami A,
Suematsu Y, Takamoto S.: Allogenicity of cryopreserved human fibroblasts: cryopreservation does not
downregulate the allogenicity of fibroblasts making
up the matrices of allografts. J Thorac Cardiovasc
Surg 120, 712-9, 2000.
20. Murakawa T, Nakajima J, Kohno T, Tanaka M,
Matsumoto J, Takeuchi E, Takamoto S. Results fromsurgical treatment for thymoma: 43 years of experience. Jpn J Thorac Cardiovasc Surg 48, 89-95, 2000.
21. Kubota H, Furuse A, Kotsuka Y, Ninomiya M, Miyaji
K, Endo M, & Takamoto S: Cardiac function evaluated by transesophageal echocardiography during cardiopulmonary bypass. Jpn J Thorac Cardiovasc Surg48, 261-266, 2000.
22. Kubota H, Takamoto S, Kotsuka Y, & Furuse A:
Successful treatment of massive pulmonary tumor
embolism from renal cell carcinoma. Ann Thorac
Surg 69, 972-3, 2000.
23. Kubota H, Takamoto S, Takeshita M, Miyaji K,
Kotsuka Y, & Furuse A: Atrial ablation using an lRK-

151 infrared coagulator in canine model. J Cardiovasc
Surg 4, 835-47, 2000.
24. Nakajima J, Takamoto S, Oka T, Tanaka M,
Takeuchi E, Murakawa T : Flow cytometric analysis
of lymphoid cells in thymic epithelial neoplasms. Eur
J Cardiothorac Surg 18, 287-92, 2000.
25. Nakajima J, Takamoto S, Tanaka M, Takeuchi E,
Murakawa T : Thoracoscopic resection of the pulmonary aspergilloma - Report of two cases. Chest
118, 1490-2, 2000.
26. Nakajima J, Takamato S, Kohno T, Ohtsuka T: Costsof videothoracoscopic surgery versus open resection
for patients with of lung carcinoma. Cancer 89, 24972501, 2000.
27. Kotsuka Y, Ezure M, Kawauchi M, Takamoto S:
Swinging motion of intimal flap through the aortic
valve in acute aortic dissection. J Cardiovasc Surg 41,
395-397, 2000.
28. Ono M., Takamoto S., Kawauchi M., Egami M.,
Kotsuka Y.: Aortobronchial fistula late after transverse arch replacement. Ann Thorac Surg 70, 964-6,
2000.
29. Suematsu Y. Ohtsuka T. Miyaji K. Murakami A.
Miyairi T. Eyileten Z. Kotsuka Y. Takamoto S. Rightheart mini-pump bypass for coronary artery bypass
grafting: experimental study. Eur J Cardio-Thoracic
Surg 18, 276-81, 2000.
30. Murakawa T, Nakajima J, Tanaka M, Karita M,
Takamoto S.: Primary low-grade B-cell lymphoma in
mucosa-associated lymphoid tissue of the lung. Jpn J
Thorac Cardiovasc Surg 49, 621-4, 2001.
31. Ohtsuka T, Suematsu Y, Kubota H, Takamoto S,
Makuuchi M. Salvage of right gastroepiploic artery
graft before pancreatoduodenectomy. J Thorac
Cardiothorac Surg 121, 1013-4, 2001.
32. Ohtsuka T, Kubota H, Motomura N, Takamoto S.
Thoracoscopy for minimally invasive axillo-coronary
artery bypass. Eur J Cardiothorac Surg 20, 856-7,
2001.
33. Ohtsuka T, Nakajima J, Kotsuka Y, Takamoto S.
Hemodynamic responses to intra-pleural insufflation
with hemipulmonary collapse. Surg Endosc 15, 132730, 2001.
34. Ohtsuka T, Takamoto S, Ono M, Motomura N.
Aortic root replacement and coronary Interposition
using a cryopreserved allograft and its branch. Eur J
Cardiothorac Surg 20, 631-2, 2001.
35. Mikio Ninomiya, Jun Nakajima, Makoto Tanaka,
Eriho Takeuchi, Tomohiro Murakawa, Takeshi
Fukami, Shinichi Takamoto: Effects of lung metastasectomy on respiratory function. Jpn J Thorac
Cardiovasc Surg 49, 17-20, 2001.
36. Mikio Ninomiya, Shinichi Takamoto, Yutaka
Kotsuka, Takeshi Miyairi, Munemoto Endo: Stentgraft implantation through partial sternotomy. J
Thorac Cardiovasc Surg 121: 992-993, 2001.
37. Mikio Ninomiya, Shinichi Takamoto, Yutaka
Kotsuka, Toshiya Ohtsuka: Indication and perioperative management for cardiac surgery in patients withliver chirrhosis: Our experience with 3 patients. Jpn J
Thorac Cardiovasc Surg 49, 391-394, 2001.
39. Mikio Ninomiya, Haruo Makuuchi, Toshiya
Ohtsuka, Shinichi Takamoto: Ischemic heart diseaseassociated with protein C deficiency. Eur J
Cardiothorac Surg 20, 883-885, 2001.
40. Mikio Ninomiya, Shinichi Takamoto, Yutaka
Kotsuka, Takeshi Miyairi, Tetsuro Morota, Hiroshi
Kubota: Midterm results after aortic valve-sparing
operation. Jpn J Thorac Cardiovasc Surgery 49, 706710, 2001.
41. Ono M., Takamoto S., Miyairi T., Kubota H.: An
extended repair of thoracic aorta through left thoracotomy after aortic root replacement. Ann Vasc Surg
15, 488-90, 2001.
42. Ono M., Wolf R.K.: Heart injury and endocarditis
caused by a needle migrated from the left arm. Eur J
Cardiothorac Surg 20, 1073-4, 2001.
43. Ueno K, Takamoto S, Miyairi T, Morota T, Shibata
K, Murakami A, Kotsuka Y: Arterial blood gas management in retrograde cerebral perfusion: the importance of carbon dioxide. Eur J Cardiothorac Surg. 20,
979-85, 2001.
44. Takeshi Miyairi, Yutaka Kotsuka, Tetsuro Morota,
Hiroshi Kubota, Ko Shibata, Yorinobu Ikeda, Tadashi
Kitamura, Takeshi Kashima, Shinichi Takamoto:Paraplegia after open surgery using endovascular
stent grafting for aortic arch. J Thorac Cardiovasc
Surg 122: 1240-3, 2001.
45. Takeshi Miyairi, Shinichi Takamoto, Yutaka
Kotsuka, Arata Murakami, Hiroshi Kubota, Toshiya
Ohtsuka: Strategy for redo surgery on the thoracic
aorta. Keio University International Symposia forLife Science and Medicine, Ed by S. Kawada, Vol. 7:
90-98, 2001. Tokyo.
46. Suematsu Y. Takamoto S. Ohtsuka T. Intraoperative
echocardiographic imaging of coronary arteries andgraft anastomoses during coronary artery bypass
grafting without cardiopulmonary bypass. J Thorac
Cardiovasc Surg 122, 1147-54, 2001.
47. Murakawa T, Nakajima J, Motomura N, Murakami
A, Takamoto S.: Successful allotransplantation of
cryopreserved tracheal grafts with preservation of
the pars membranacea in nonhuman primates. J
Thorac Cardiovasc Surg 123, 153-60, 2002.
48. Kubota H, Takamoto S, Murakami A, Kotsuka Y, &Furuse A: Biologic glue-molded “bio-mini-paddle”
electrode used for atrial and ventricular pacing anddefibrillation. Ann Thorac Surg 322-4, 2002.
49. Kotsuka Y, Ezure M, Tanaka K, Takamoto S,
Kuwahara K, Ikeda H. Ultrathin-wall vascular grafts
for endovascular surgery. J Art Org 5: 108-112, 2002．
50. Suematsu Y. Ohtsuka T. Miyairi T. Motomura N.
Takamoto S. Ultrasonic evaluation of graft anastomoses during coronary artery bypass grafting without cardiopulmonary bypass. Ann Thorac Surg 74,
273-5, 2002.

227

Department of
Gastrointestinal Surgery /Department of Surgical
Metabolism and Nutrition and
Endocrine Surgery
Outline and Research Objectives
Since 2001, the former Third Department of Surgery, which was located in a branch hospital of the University of Tokyo, has been divided into two departments, the Department of
Gastrointestinal Surgery and the Department of Surgical Metabolism and Nutrition and
Endocrine Surgery, in line with the integration of the main and branch hospitals the elevation to a department in the graduate school of medicine at our university. Our research
activities in both departments have been well organized and ultimately successful by maintaining a close connection.
The main research activities of the department of Gastrointestinal Surgery are focusedon diagnosis and therapy for gastrointestinal diseases and clinical and basic research forgastrointestinal carcinogenesis. A historical and outstanding achievement of the department
of Gastrointestinal Surgery is the successful innovation of the “Gastrocamera” a half century
ago. Thereafter, we have been active in the diagnosis and treatment of gastric cancer andestablished the feasibility of extended gastrectomy with intraoperative chemotherapy foradvanced cancer, and limited lymph node dissection for early gastric cancer. With a view tofurther development of research for gastrointestinal carcinogenesis, we established theJapanese Society for Gastroenterological Carcinogenesis in 1989, and have been studying
the underlying mechanisms of development, progression and prevention of digestive cancer.
The department of Surgical Metabolism and Nutrition and Endocrine Surgery has been
studying one of the most fundamental issues in surgery, i.e., “surgical stress,” which indicates postoperative physiological and endocrinological internal reaction, and nutritional support for the postoperative patients. Our department is a pioneer in this area in Japan, andwe established the Japanese Society for Surgical Metabolism and Nutrition in 1965. In addition, we have been studying endocrine issues, i.e., surgical therapy for breast, thyroid andparathyroid diseases.

Faculties and Students

Past Research and Major Accomplishments

Professor and Chairman Michio Kaminishi, MD., Ph.D.
(since 1997)
Associate Professor Ken-ichi Mafune, MD., Ph.D.
(Yoshikazu Mimura, M.D., Ph.D.
from Division of Surgical
Operation Center)
Lecturer
Shouji Shimoyama, MD., Ph.D.
Toshihisa Ogawa, MD., Ph.D.,
Mitsue Saito MD., Ph.D.,
Associate ...............................12
Postdoctor Fellow ...................8
Graduate Student....................9
Secretary .................................4

In research on gastrointestinal carcinogenesis, wehave established experimental models of gastric carcinogenesis and looked closely at the important rolesof repetition of injury and regeneration of gastricmucosa. In particular, we demonstrated that infectionof Helicobacter pylori acts not only in promotion but
also in co-initiation of gastric carcinogeneis. This was
the first report in the world to show the direct relationship between H.p. infection and gastric carcinogenesis using experimental models. Furthermore, we
verified that duodenogastric reflux and denervation of
the gastric mucosa enhance gastric mucosal injury,
resulting in development of gastric remnant cancer.

228

Based on these results, we have adopted new operations for gastric cancer and obtained better results interms of postoperative QOL and outcome. Ourdetailed histopathological analysis of colon carcinogenesis revealed de novo carcinogenesis other thanadenoma-carcinoma sequence, leading to the current
hypothesis of the dysplasia-carcinoma sequence.
In research on surgical metabolism and nutrition,
we clearly showed the importance of nutritional support under condition of postoperative hypoxia andendotoxemia. Fatty acids reduced the morbidity andmortality due to panperitonitis, and glutamine attenuated acute lung injury after the organism was challenges with endotoxin. Research on the metabolismof phosphate, calcium and sodium is one of the mostimportant means to clarify the mechanism of the
postoperative physiological reaction. We demonstrated the close relationship between these metabolites
and catecholamine after surgical stresses such as
hypoxia, endotoxiemia and ischemic reperfusion
injury. These studies aim to reduce the intra- andpost-operative stresses that would be risky forpatients.

Current Research
The main focus of our current research in the fieldof Gastrointestinal Surgery is the tailor-made treatment of cancer based on its stage and characteristics.
We try to establish the optimal limited surgery forearly cancer by application of sentinel node navigation system and molecular analysis of lymph node
micrometastasis. For advanced cancer, multimodal
therapy combined with extended surgery and perioperative chemotherapy is required. In regard tochemotherapy, a new strategy using apoptosis-related
and/or cell differentiation-related agents is likely toachieve better prognosis in advanced cases.
Current research topics in the department of
Gastrointestinal Surgery are the followings.
1) Carcinogenesis of gastrointestinal cancer･ Roles of Helicobacter pylori infection in gastriccarcinogenesis･ Roles of Trefoil Peptides in gastric metaplasia･ Clinical and experimental studies on Barrett
esophagus
2) Molecular mechanisms of gastrointestinal cancer･ Effects of hypoxia and nutritional deprivation on
cancer development and progression･ Roles of PPARr in gastric carcinogenesis and treatment of gastric cancer by PPARr ligands･ Angiogenic factors in gastrointestinal cancer･ Telomerase activity in gastrointestinal tumors･ Lymph node micrometastasis of gastric cancer

3) Minimally invasive treatment of upper GI cancers･ Indication and results of laparoscopic surgery･ Sentinel node navigation surgery for early gastriccancer･ Evaluation of results of PPG and jejunal interposition after gastrectomy in terms of postoperative
QOL4) Multimodal treatment for esophageal and gastric cancers･ Neoadjuvant or definitive chemoradiation therapy
for esophageal cancer･ Apoptosis induced by TRAIL･ Prediction of gastric cancer recurrence by molecular analysis of samples of peritoneal washes･ Mechanisms of adverse effects of chemotherapy
for gastrointestinal cancer on the intestinal
mucosa and its preventive therapy5) Gastrointestinal motility･ Mechanism of peppermint oil solution on relaxation of digestive tract･ Role of cytokine and COX-2 in gastrointestinal
motility･ Effects of intra- and extra-abdominal environments on gastrointestinal motility･ Gastric motility after gastrectomy
The main focus of our current research in the fieldof Surgical Metabolism and Nutrition is “adaptive
response to surgical stresses”. In particular, “cross tolerance among different stresses” is a very challenging
and important phenomenon to be elucidated, and theclinical application of findings will lead to reductionof morbidity and mortality due to severe surgicalstresses and endotoxin shock. On the other hand,
there is a definite difference in adaptive response tosurgical stress between genders. We have clearly
demonstrated the gender difference in cytokine secretion after endotoxin challenge.
In the field of Endocrine Surgery, we have introduced minimally invasive surgery for breast cancer
and thyroid tumors through the application of laparoscopic procedures. In terms of differential diagnosis of
thyroid tumors, we established a method for the
immunohistochemical detection of telomerase
reverese ranscriptase (hTERT) by in situ hybridization, providing a good marker for distinction between
follicular adenoma and follicular cancer.
Current research topics in the department ofSurgical Metabolism and Nutrition and Endocrine
Surgery are the followings2291) Surgical metabolism and nutrition･ Mechanisms of cross tolerance among different
stresses (endotoxin-hypoxia/ hypoxia-endotoxin)
after surgery･ Role of catecholamines in adaptation to surgicalstress such as endotoxemia･ Bacterial translocation after major surgery andanti-cancer chemotherapy･ Gender difference is a modulation factor for postoperative morbidity･ Parathyroid function after surgical stress･ Ischemic preconditioning and its underlying
mechanism (NO, HSP, metalothionein)
2) Endocrine surgery･ Cytology of breast and thyroid tumors by in situ
hybridization of telomerase reverse transcriptase(hTERT)･ Effects of preoperative endocrine therapy of breast
cancer･ Detection of breast cancer cell in the drainage vein
from the breast by using RT-PCR･ Molecular analysis of heterogeneity in breast andthyroid cancers･ Development of QOL questionnaire for breast cancer patients

Future Prospects (Figure 1)
Our main theme in basic and clinical research is“Surgery and Inflammation”. There are 2 phases of230

inflammation, acute and chronic. Under acute inflammation or acute stress, injured tissue recovers to normal promptly after subsidence of the acute causes. In
that case, the organ represses apoptosis and maintains or restores the normal tissue. Many factors suchas cytokines, NO, NF-κB, and HSP play importantroles in development and progression of the inflammatory process. Conversely, these molecular factors
play a key role in the attenuation of morbidity
through across tolerance mechanism. Clinical application of therapy based on cross tolerance leads to a better prognosis for morbid patients after severe surgicalstress.
Under chronic inflammation, for, example, In
cases of chronic gastritis, however repetition of injury
and subsequent regeneration is observed regardless ofthe causes. The accelerated regeneration of the gastricmucosa induces a newly developed tissue that is
adapted for chronic injury by balancing cell proliferation and apoptosis. This phenomenon is widely recognized as metaplasia and all metaplastic change in thehuman organ is considered to be a result of adaptive
reaction. Furthermore, the metaplastic cells and/or thecells departing from the adaptive process have potential for malignant transformation.
The relation between chronic inflammation andcarcinogenesis is commonly observed not only in thestomach, but also in the esophagus, colon, liver andother tissues. Examples include reflux esophagitis andBarrett’s cancer, ulcerative colitis and colitic cancer,
viral hepatitis and hepatoma. Some key factors suchas NF-kB, COX, NO, PPARr, HSP, and trefoil peptides
play important roles in the process of chronic inflammation. In addition, those factors are also expressedin breast and thyroid tumors.
The development of molecular biology-based diagnosis for cancers and the tailor-made treatment forcancer patients depending on the properties of eachcancer and each patient’s genetic background, areurgent issues. Further analysis of the relevant factors
may contribute to advances in diagnosis, treatment
and prevention of not only gastrointestinal cancers
but also breast and thyroid cancers. In particular,
PPARr, trefoil peptide and histone deacetylase
inhibitors (HDAIs) modulate not only cell proliferation but also cell differentiation. We are trying todevelop a new strategy of molecular targeting therapy
for gastrointestinal, breast and thyroid cancers byapplication of these cell-differentiating factors.

Research Grants
･
･

･

･
･

Clonal analysis of isolated intestinal metaplastic
glands of stomach using X linked polymorphism
Quantitative detection of micrometastases in thelymph nodes of gastric cancer patients with real-time
RT-PCR
Histopathological change and expression of genes
and proteins in chief cells through gastric carcinogenesis in rodent models
Sublethal endotoxin administration evokes superresistance to systemic hypoxia.
Telomeres and telomerase activity in digestive andendocrine organs: Clinical application to the diagnosis and therapy.

Select Publications
1. Kaminishi M, Sadatsuki H, Johjima Y, Oohara T,
Kondo Y. A new model for production of chronic
gastric ulcer by duodenogastric reflux in rats.
Gastroenterology 92, 1913-1918,1987.
2. Mimura Y, Yamakawa M, Furuya K, Oohara T. Effectof oxygen supply on protein metabolism in surgicallyinjured rats-oxygen as a nutrient. Ann Surg 212,228233,1990.
3. Maeda J, Dudrick SJ. Rapid spectrophotometricdetermination of plasma carnitine concentrations.
JPEN 14, 527-532,1990.
4. Mimiura Y, Khraibi AA, Heublein DM, Knox FG.
Acute hypoxia blunts natriuretic and diuretic effcets
of atrial natriuretic factor in rats. Am J Physiol 266,103-1509,1994.
5. Shimoyama S, Kaminishi M, Joujima Y, Oohara T,
Hamada C, Teshigawara W. Lymph node involvement correlation with survival in advanced gastriccarcinoma: Univariate and multivariate analysis.
Journal of Surgical Oncology 57, 164-170,1994.

6. Kaminishi M, Shimizu N, Shimoyama S, Yamaguchi
H, Ogawa T, Sakai S, Kuramoto S, Oohara T.
Etiology of gastric remnant cancer with special reference to the effects of denervation of the gastricmucosa. Cancer 75, 1490-1496,1995.
7. Hiki N, Berger D, Buttenschoen K, Boelke E,
Seidelmann M, Strecker W, Kind L, Beger HG.
Endotoxemia and specific antibody behaivior against
different endotoxins following multiple injuries. J
Trauma 38, 794-801,1995.
8. Kaminishi M, Shimizu N, Nomura S, Ohkura F,
Shimoyama S, Yamaguchi H, Hashimoto M, Ogawa
T, Sakai S, Oohara T. Different carcinogenesis in thegastric remnant following gastrectomy for gastriccancer. Cancer 77, 1646-1653, 1996.
9. Nomura S, Kaminishi M, Sugiyama K, Oohara T,
Esumi H. Clonal analysis of isolated single fundic
and pyloric gland of stomach using X -linked polymorphism. Biochem Biophy Res Commun 226,385390,1996.
10. Nakamura T, Yamakawa M, Maeda J, Mimura Y,
Yamada H, Ogawa T, Kanauchi H, Oohara T. Effectof glutamin on acute lung injury in rats with endotoxemia. Clinical Nutrition 16,79-83,1997.
11. Kanauchi H, Mimira Y. Adrenalectomy overcomes
the blunted natriuretic response to atrial natriuretic
peptide during hypocapnia in rats. Clin Exp
Pharmacol Physiol 24,850-855,1997.
12 Kuramoto S, Mimura T, Yamazaki K, Kobayashi K,
Hashimoto M, Sakai S, Kaminishi M, Oohara T. Flat
concers do develop in the polyp-free large intestine.
Dis Colon Rec 40,534-542,1997.
13. Shimizu N, Kuramoto S, Mimura T, Kobayashi K,
Kobayashi M, Sakai S, Kaminishi M, Oohara T.
Helicobactor pylori promotes development of
pepsinogen altered pyloric glands, a prreneoplastic
lesion of glandular stomach. Cancer Letter 27, 546549,1997.
14. Kaminishi M, Yamaguchi H, Shimizu N, Nomura S,
Yoshikawa A, Hashimoto M, Sakai S, Oohara T.
Stomach-partitioning gastrojejunostomy for unresectable gastric carcinoma. Arch Surg 132,184187,1997.
15. Nomura S, Kaminishi M, Sugiyama K, Oohara T,
Esumi H. Clonal analysis of isolated intestinal metaplastic gland of stomach using X-linked polymorphism. Gut 42. 663-668, 1997.
16. Hiki N, Beger D, Boelke E, Nussler A, Seidelmann M,
Braumueller HD, Oohara T, Beger HG. Endotoxin
elimination by human monocytes-endotoxin is neutralized via CD14? Shock 7,22-23,1997.
17. Mimura Y, Yamakawa M, Maeda J, Tateno I, Araki S,
Fujita T, Sugizaki K, Furuya K, Oohara T. Efficacy ofamino acid infusion for improving protein metabolism after surgery-A prospective randomized study inpatients undergoing subtotal gastrectomy. J Am Coll
Surg 185,163-171,1997.
18. Hiki N, Berger D, Boelke CPE, Wiedeck H,
Seidelmann M, Staib L, Oohara T, Kaminishi M,
Beger HG. Endotoxin binding and elimination by231monocytes: the secretion of soluble CD14 represents
an inducible mechanism counteracting reduced
expression of membrane CD14 during sepsis and ina patient with paroxysmal nocturnal hemoglobinuria. Infect Immun 66, 1135-1141,1998.
19. Kanauchi H, Yamasaki K, Ogawa T, Mimura Y.
Endoscopic thyroidectomy in a porcine. Endocrine J
45,135-136,1998.
20. Sugiyama A, Maruta F, Ikeno T, Ishida K, Kawasaki
S, Katsuysma T, Shimizu N, Tatematsu M.
Helicobacter pylori infection enhances MNU (Nmethyl-N-nitrosourea)-induced stomach carcinogenesis in the Mongolian Gerbil.Cancer Res 58, 20672069,1998.
21. Zhang D, Shimoyama S, Kaminishi M. Feasibility ofpylorus-preserving gastrectomy with a wider scopeof lymphadenectomy. Arch Surgery 133,993997,1998.
22. Shimizu N, Inada K, Nakanishi H, Tsukamoto T,
Ikehara Y, Kaminishi M, Kuramoto S, Sugiyama A,
Katsuyama T, Tatematsu M. Helicobacter pylori
infection enhances glandular stomach carcinogenesis
in Mongolian gerbils treated with chemical carcinogens. Carcinogenesis 20, 669-676,1999.
23. Shimoyama S, Shimizu N，Kaminishi M. Type-oriented intraoperative and adjuvant chemotherapy andsuvival after curative resection of advanced gastriccancer. World J Surg 23,284-292, 1999.
24. Shimoyama S, Yamasaki K, Kawahara M, Kaminishi
M. Increased angiogenin concentration in colorectal
cancer is correlated with cancer progression. ClinicalCancer Res 5,1125-1130, 1999.
25. Mafune K, Tanaka Y. Influence of multimodality
therapy on the cellular immunity of patients withesophageal cancer. Ann Surg Oncol 7,609-616, 2000.
26. Nakamura K, Furugori E, Esaki Y, Arai T, Sawabe M,
Okayasu I, Fujiwar M, Kammori M, Mafune K, Kato
M. Correlation of telomere lengths in normal andcancers tissue in the large bowel. Cancer Lett
158,179-184, 2000.
27. Shimizu N, Ikehara Y, Inaba K, Nakanishi H,
Tsukamoto T, Nozaki K, Kaminishi M, Kuramoto S,
Sugiyama A, Katsuyama T. Eradication Diminishes
Enhancing Effects of Hellicobactor pylori Infection
on Glandular Stomach Carcinogenesis in Mongolian
Gerbils. Cancer Res 60, 1512-1514, 2000.
28. Shimoyama S, Kaminishi M. Increased angiogenin
expression in gastric cancer correlated with cancer
progression. J Cancer Res Clin Oncol 126, 468-474,
2000.
29. Seto Y, Takenoue T, Nariko H, Kaminishi M.
Influence of gastrectomy on the distribution of intravenously administered 5-fluorouracil to regional
lymph nodes in an animal model. Cancer 88,24432446, 2000.
30. Shimoyama S, Aoki F, Murakami T, Toyoda K,
Kaminishi M. The feasibility of daily concurrent
chemoradiation therapy as a nonsurgical management for esophageal cancer: Our experience and the-232

oretical backgrounds. J Surg Oncol 74, 134-137,
2000.
31. Shimoyama S, Seto Y, Yasuda H, Kaminishi M.
Wider indications for the local resection of gastriccancer by adjacent lymphadenectomy. J Surg Oncol,
75, 157-164, 2000.
32. Takahashi K, Saito M, Makita M, Tada T, Uchida Y,
Yoshimot M, Kasumi F, Akiyama F, Sakamoto G.
Surgery for ductal carcinoma in situ. Breast Cnacer
7,337-340, 2000.
33. Hiki N， Berger D， Mimura Y， Frick J, Dentener
MA， Buurman WA，Seidelmann M，Kaminishi
M， Beger HG. Endotoxin binding proteins during
major elective surgery: role of sCD14 for phagocytic
activation. World J Surg 24,499-506, 2000.
34. Kanauchi H, Mimura Y, Hiki N, Kaminishi M.
Catecholamine and cytokine response to laparoscopic adrenalectomy in patients with pheochromocytoma. Biomed Pharmacother 54,191-193, 2000.
35. Kammmori M, Takubo K, Nakamura K, Furugouri E,
Endo H, Kanauchi H, Mimura Y, Kaminishi M.
Telomerase activity and telomere length in benign
and malignant human thyroid tissues. Cancer Lett
159,175-181, 2000.
36. Ogawa T, Mimura Y, Hiki N, Kanauchi H, Kaminishi
M. Ischaemic preconditioning ameliorates functional
disturbance and impaired renal perfusion in rat
ischaemia-reperused kidneys. Clin Experimental
Pharmacol Physiol 27,997-1001, 2000.
37. Ogawa T, Nussler AK, Tuzuner E, Neuhaus P,
Kaminishi M, Mimura Y, Beger HG. Contribution of
nitric oxide to the protective effects of ischemic preconditioning in ischemia-reperfused rat kidneys. J
Lab Clin Med 138, 50-58, 2001.
38. Imamura K, Ogura T, Kishimoto A, Kaminishi M,
Esumi H. Cell Cycle Regulation via p53
Phosphorylation by a 5’-AMP Activated Protein
Kinase Activator, 5-Aminoimidazole-4-Carboxamide1-β-Ribofuranoside, in a human Hepatocellular
Carcinoma Cell Line. Biochem Biophy Res Commun
287, 562-567, 2001.
39. Kammori M, Kanauchi H, Nakamura K, Kawahara M,
Weber T, Mafune K, Kaminishi M, Takubo K.
Demonstration of human telomerase reverse transcriptase in human colorectal carcinomas by in situ
hybridization. Int J Oncol 20,15-21, 2001.
40. Seto Y, Yamaguchi H, Shimoyama S, Shimizu N,
Aoki F, Kaminishi M. Results of local resection with
regional lymphadenectomy for early gastric cancer.
Am J Surgery 182, 498-501, 2001.
41. Kawahara M, Kammori M, Kanauchi H, Noguchi, C
Kuramoto S, Kaminishi M. Immunohistochemical
prognostic indicators of gastrointestinal carcinoid
tumors. Eur J Surg Oncol 28, 140-146, 2001.
42. Asai K, Hiki N, Mimura Y, Ogawa T, Unou K,
Kaminishi.M. Gender differences in cytokine secretion by human peripheral blood mononuclear
cell:role of estrogen in modulating lps-induced
cytokine secretion in an EX VIVO septic model.
Shock16, 340-343, 2001.

43. Kammori M, Mafune K, Kawahara M, Mimura Y,
Kaminishi M, Takubo K. Telomere shortening withaging in human thyroid and parathyroid tissue.
Experiment Gerontol 37,513-521, 2002.
44. Kammori M, Nakamura K, Kamnauchi H, Obara T,
Kawahara M, Mimura Y, Kaminishi M, Takubo K.
Consistent decrease in telomere lemgth in parathyroid tumors but alteration in telomere activity limited to malignancies: Preliminary report. World J Surg
26,1083-1087, 2002.
45. Nozaki K, Shimizu N, Tsukamoto T, Inada K, Cao X,
Ikehara Y, Kaminishi M, Sugiyama A, Tatematsu M.
Reversibility of Heperotopic Proliferative Glands in
Glandular Stomach of Helicobacter pylori-infected
Mongolian Gerbils on Eradication. Jpn J Cancer Res
93,374-381, 2002.
46. Li QL, Ito K, Sakakura C, Fukumachi H, Inoue K, Chi
XZ, Kaneda A. Causal relationship between the loss
of RUNX3 expression and gastric cancer. Cell
109,113-124, 2002.
47. Kaneda A, Kaminishi M, Nakanishi, Y Sugimura T,
Ushijima T. Reduced Expression of the InsulinInduced Protein 1 and p41 ARP2/3 Complex Genes in
Human Gastric Cancers. Int J Cancer 100, 57-62,
2002.
48. Yamaguchi H, Goldenring JR, Kaminishi M, Lee JR.
Identification of Spasmolytic Polypeptide Expressing
Metaplasia(SPEM) in Remnant Gastric Cancer andSurveillance Postgastrectomy Biopsies. Dig Dis
Science 47,573-578, 2002.
49. Shimoyama S, Kaminishi M. Supra-additive antitumor activity of 5FU with tumor necrosis factor related apoptosis inducing ligand on gastric and coloncancers in vitro. Int J Oncol 21,643-648, 2002.
50. Yamaguchi H, Goldenring JR, Kaminishi M, Lee JR.
Association of spasmolytic polypeptide-expressing
metaplasia with carcinogen administration andoxyntic atrophy in rats. Lab Invest 82:1045-1052,
2002

233

Department of Hepato-BiliaryPancreatic Surgery /Department of Artificial
Organ and Transplantation
Outline and Research Objectives
In September 1893, Professor Sankichi Satoh founded “the Second Department ofSurgery, University of Tokyo”. He was a pioneer of visceral surgery in Japan and he also
founded the Japanese Society of Surgery in 1899. His pupils, professor Waichiroh Okada,
professor Keizo Doi, professor Hisashi Ishihara founded departments of otorhinolaryngology, dermatology, and dental surgery, respectively at Unversity of Tokyo Hospital.
In May 1952, professor Seiji Kimoto became the 5th chairman of our department. He wasa pioneer of cardiovascular surgery in Japan, and founded the department of thoracic surgery in 1964. In the same year, he also performed the first case of kidney transplantation in
Japan.
In July 1971, professor Shigeru Hatano, the 6th chairman, founded the department ofPediatric Surgery.
In April 1994, I (professor Masatoshi Makuuchi) became the 10th chairman. I started living donor liver transplantation on January 31st, 1996 and has done 191 cases by the end of
September 2002. As the result of the shift to the Graduate School of Medicine, the three
Departments of Surgery were transformed to one Department and 6 divisions in April 1998.
Our second Department of Surgery took charge of “Hepato-Biliary-Pancreatic Surgery
Division, andArtificial Organ and Transplantation Division”.
Our current main research objectives are surgical treatment of hepato-biliary-pancreaticcancer, liver transplantation, and development of artificial liver.

Faculties and Students
Professor and Chairman Masatoshi Makuuchi, MD,
PhD (1994-)
Associate Professors Norihiro Kokudo, MD
Yasuhiko Sugawara, MD
Lecturer
Hiroshi Imamura, MD
Associate ...............................11
Postdoctal Fellow....................5
Graduate student ....................9
Research student.....................2
Secretary .................................4

Past Research and Major Accomplishments
Between late 1970’s and early 1980’s, I have developed the application of ultrasound to the various
treatment modalities of hepatobiliary diseases. First, I
have developed the percutaneous transhepatic cholangiography (PTC) and the percutaneous transhepatic
biliary drainage (PTBD) method under ultrasoundguidance. With these methods, intrahepatic biliary
tract was punctured selectively and drainaged directly
by real-time guidance. Ultrasound-guided PTC and

234

PTBD methods are simple and easy to perform andnow are widely used over the world. I also investigated precisely the anatomy and its variation of theintrahepatic portal branches for the first time by theultrasound examination. Before this work, information on its anatomy was obtained mainly by the studyof the cadaveric liver which was somewhat mal
understood or by the angiographic studies which byitself does not allow a precise evaluation on eachpatients. I then developed an intraoperative ultrasound probe and it was facilitated liver surgery for the
first time in the world. By its usage, the positional
relationship between the tumor and intrahepatic vasculobiliary structure can be understood during thehepatic parechymal transection. This is particularly
important in liver resection in cirrhotic liver because
the examination of the tumor by bimanual palpation
is difficult in such cases. Further, I developed themethod to resect anatomically the Couinaud’s segment by the aid of ultrasound and so-called staining
technique of the intrahepatic portal branch. This
small but anatomical resection is now accepted as astandard procedure for the patients with hepatocellular carcinoma de facto and called also as ‘Makuuchi’sprocedure’. Based on the findings of the liver anatomy, I have developed and performed, besides theabove described anatomical resection, central bisectorectomy or right paramedian sectorectomy. This canbe considered a remarkable advance in the era whensolely the right or left hepatcetomy was performed as
anatomical hepatc resections.
Development and application of intermittent
hemihepatic or total inflow occlusion technique is alsoa fruit of my effort in this period. In 1980’s, total
inflow occlusion was sporadically carried out over theworld. But it was used always as a continuous mannerand maximum length of the occlusion was considered
60 minutes. Moreover, it was almost a central dogmathat the cirrhotic liver could not tolerate the inflow
ischemia. Since intermittent ischemia was a recycle ofischemia/reperfusion injury it was believed to beworse than continuous ischemia. In these circumstances, I, for the first time, proved the inflow occlusion can be applied to cirrhotic liver and intermittent
inflow occlusion can be applied safely and better thancontinuous occlusion technique.
The development of various operative procedures
preserving the thick short hepatic vein as a drainage
vein of right lateral sector and scarifying the righthepatic vein is the achievement of this period. In
addition, I first demonstrated the right hepatic veincan be safely divided and ligated extrahepatically if
the inferior vena cava ligament was divided and cut
prior to the dissection. These vein and ligament are
now called as ‘Makuuchi’s vein’ and ‘Makuuchi’s ligament’ worldwide.
Extended right hemihepatectomy is often necessary in the surgical treatment of hilar cholangiocarcinoma. Such a procedure may lead to the increased
risk of postoperative liver insufficiency. I have developed and applied the preoperative portal vein
embolization (PVE) to induce the atrophy of the liver
to be resected and thus induce the compensatory
hypertrophy of the liver part to be preserved after theresection. In 1982, I performed the first case of PVE inthe world. This method is simple to apply and now is
widely applied all over the world.
By virtue of the development of these procedures
and improvement in the technical skill, the operative
mortality of liver resection in my institute is almost
zero including that for the cirrhotic liver and for thehilar cholangiocarcinoma compromized with obstructive jaundice.
In June 1990, I started living donor liver transplantation (LDLT). Based on my own experience of LDLT,
I have proposed a formula that allows the calculationof the ideal liver volume to the recipient or the donor
used the preoperative decision of what kind of donor
hepatectomy should be performed. This formula is
now called as ‘Makuchi’s formula’. In 1993, I per-formed successfully for the first time in the world theadult-to-adult LDLT using left hemiliver. Following
this success, I have carried out adult-to-adult LDLT
including emergency transplantation for patients withfulminant hepatic failure. The number of patients
undergoing LDLT until now exceeds 200 cases withless than 10 % overall mortality. Recent my contribution to this field is the establishment of the feasibilityof the right lateral sector graft and the establishmentof the criteria for reconstruction of the middle hepatic
venous tributaries in case of right hemiliver graft.

Current Research
I have established a safe and effective hepatic
resection techniques based on the liver anatomy andchallenged repeated hepatic resections for recurrent
liver tumors including hepatocellular carcinoma (HCC)and metastatic tumors. As one of the top institutes ofhepato-biliary surgery in the world, we have achieved
no mortality and minimum morbidity hepatic resection in recent series of patients, and we have encouraged surgeons worldwide. The accumulated clinicaldata will reveal the long-term results that demonstrate the advantages of surgical treatment for liver
tumors. The molecular biological investigations ofliver tumors will elucidate the biological behaviors
and prognostic factors of these malignant diseases.
However, there are a number of patients with HCC
who are not candidates for hepatic resection becauseof the limited liver functional mass. Since Milan groupreported promising results of orthopic liver transplantation for HCC in selected cases, I have started living
donor liver transplantation (LDLT) for recurrent HCC.
As one of the treatment options for HCC, LDLT will
be commonly performed in Japan. The indications for
cadaveric liver transplantation for HCC have been limited because of donor shortage. I am trying to expand
the indications of LDLT for HCC beyond the Milancriteria. The long-term results may clarify the indications for LDLT for HCC.
Strong and co-workers reported the first successfultransplantation from a living donor (LDLT). LDLT ismainly conducted in an attempt to alleviate the shortage of donor organs and to decrease mortality amongchildren awaiting transplants. I started the LDLT program in 1990 when I was in Shinshu University. The
indications of LDLT expanded to the adults and I performed the first successful case in 1994. After I
moved to University of Tokyo, I started the programin 1996. Until September 2002, I accumulated 191
cases. The mortality rate of the patients is less than
10%, which is much superior to the world standard.
The major limitation for LDLT for adults is theadequacy of the graft size. To overcome this problem,
I devised the left liver plus caudate lobe graft with its

235

venous reconstruction. The caudate lobe can provide
about a 7% increase in graft weight, which contributed
the less frequent cholestasis after the operation. The
right liver graft is now most commonly used graft in
LDLT for adults. The right liver graft is usually harvested without the trunk of the middle hepatic vein.
However, this type of grafts can cause severe congestion of the right paramedian sector because middle
hepatic vein often drains the considerable part of this
sector. Middle hepatic vein drainage into the recipient’s venous system can be reconstructed using vein
grafts. This provides a functioning liver mass comparable to an extended right liver graft. I firstly proposed reconstruction criteria.
Right liver harvesting from living donors is sometimes dangerous when the volume ratio of right liveris large. Some donors have larger right lateral sector
compared with left liver. I firstly performed LDLT
suing this type of graft.

List of On-going Research Projects
1) Hepato-pancreato-biliary surgery
Extended hemihepatectomy for hilar bile duct carcinoma
Development of ultrasonic diossector equippedwith electrocautery
Liver resection of liver metastases from GIST
Long-term evaluation of two-staged pancreaticoduodenectomy
Intraoperative ultrasound-guided direct pancreatography
Liver hypertrophy after hepatopancreatoduodenectomy
Resection of S4inf during hepaticojejunostomy
Reconstruction of superior mesenteric vein using
cryopreserved homograft vein
Repeated hepatic resection for colorectal metastases
Risk factors for biliary leakage after liver resection
Arterial buffer response in cirrhotic patients
Evaluation of hepatic functional reserve using GSA
scintigraphy
Resection of IVC wall and/or root of hepatic veinin colorectal metastasis
Effect of cimetidine on the postoperative recurrence in HCC patients
Disruption of pRb-p16 INK4 pathway: a common
event in ampullary carcinogenesis
Qualitative and quantitative analysis of desgamma-carboxy prothrombin (DCP) in cancerous
and non-cancerous liver tissue of patients with
HCC
Protein induced by Vitamin K absence or antagonist II absence (PIVKA-II) in cancerous and non-236

cancerous liver tissue of patients with HCC
Serum anandamide levels in infection
2) Transplantation
Diagnostic criteria for fungal infection in posttransplant patients
Quality of life in donors for LDLT
Argorythms for the timing of liver biopsy after
LDLT
Effect of hANP in post-transplant oliguria
Serological marker for early detection of carinii
pneumonia
Reconstrucrion of venous tributaries in right livergraft (long-term outcome)Re-evaluation of the Milan’s criteria for the indication of cadeveric livertransplantation to patients with hepatocellular carcinoma
Donor inflow occlusion technique in living donorliver transplantation
Liver regeneration in donors
Development of anti-septic biliary drainage tube
for LDLT
3) Artificial Organ
Artificial liver for xeno-cross circulation and clinical application
Artificial liver and transgenic pig for clinical application
Development of artificial erythrocyte for liver support system
Reconstruction of liver tissue using bone marrowtransplantation

Future Prospects
Beyond the technical challenges are various medical issues that perhaps need to be given greater attention in the future. LDLT has come to address a healthconcern of epidemiologic dimension in Japan, namely
hepatitis B and C and consequently occurred hepatocellular carcinoma. Recurrent disease after liver transplantation remains a major threat and efforts aimed atprevention and cure have to continue.
Issues related to immunosuppression have to be
tackled and induction or facilitation of graft tolerance,
which may not be as elusive in LDLT, is yet to beachieved. Management of infection in the immunosuppressed transplant patient is a peculiar challengethat transplant specialists must face in the near
future.
Research project of artificial liver device is almost
close to clinical use for the bridge to liver transplantation for the patients with liver failure. Recent advanceof research about the regeneration medicine has
demonstrated the possibility of hepatocyte differenti-

ation from bone marrow or embryonic stem cells. For
the treatment of fulminant liver failure, the humanhepatocytes cultured and expanded from bone marrow will be used in the future.

Research Grants
1999-2001 Improvement of surgical treatment for biliary
cancer.
Grant for the prevention of cancer, the ministry ofhealth and labor
4,800,000 yen
1999-2001 Artificial liver for xeno-cross circulation andclinical application
Grant from the ministry of education and science(A)(2)
35,800,000yen
2000-2001 Clinical improvement and development oforgan transplantation
Grant for the human genome and regeneration, theministry of health and labor
4,500,000 yen
2000-2001 Research on intractable liver disease
Grant for intractable diseases, the ministry of healthand labor
1,000,000 yen
2001-2002 Artificial liver and transgenic pig for clinicalapplication
Grant for advanced medicine (A), the ministry ofhealth and labor
196,840,000 yen

Select Publications
1. Makuuchi M, Bandai Y, Ito T, Wada T: Ultrasonicallyguided pancreatography. Radiology 134:767-770,
1980.
2. Makuuchi M, Bandai Y, Ito T, Watanabe G, Wada T,
Abe H, Muroi T: Ultrasonically guided percutaneoustranshepatic bile drainage: A single step procedure
without cholangiography. Radiology 136:165169,1980.
3. Makuuchi M, Hasegawa H, Yamazaki S, Bandai Y,
Watanabe G, Ito T: The inferior right hepatic vein :ultrasonic demonstration. Radiology 148:213-217,
1983.
4. Makuuchi M, Hasegawa H, Yamazaki S Moriyama N,
Takayasu K, Okazaki M: Primary Budd-Chiari syndrome: ultrasound demonstration. Radiology
152:775-779, 1984.
5. Makuuchi M, Sukigara M, Mori T, Kobayashi J,
Yamazaki S, Hasegawa H, Moriyama N, Takayasu K,
Hirohashi S: Bile duct necrosis: complication of transcatheter hepatic arterial embolization. Radiology
156:331-334, 1985.
6. Makuuchi M, Hasegawa H, Yamazaki S:
Ultrasonically guided subsegmentectomy. Surgery,
Gynecology & Obstetrics 161:346-350, 1985.
7. Gunven P, Makuuchi M, Takayasu K, Moriyama N,
Yamasaki S, Hasegawa H: Preoperative imaging ofliver metastases - comparison of angiography CT
scan and ultrasonography. Annals of Surgery
202:573-579, 1985.
8. Makuuchi M, Hasegawa H, Yamazaki S, Takayasu K:
Four new hepatectomy procedures for resection of
the right hepatic vein and preservation of the inferior
right hepatic vein. Surgery, Gynecology & Obstetrics
164:68-72, 1987.
9. Makuuchi M, Mori T, Gunvén P, Yamazaki S,
Hasegawa H: Safety of hemihepatic vascular occlusion during resection of the liver. Surgery,
Gynecology & Obstetrics 164:155-158, 1987.
10. Makuuchi M, Hasegawa H, Yamazaki S, Takayasu K,
Moriyama N: The use of operative ultrasound as an
aid to liver resection in patients with hepatocellular
carcinoma. World Journal of Surgery 11:615-621,
1987.
11. Sakamoto M, Hirohashi S, Tsuda H, Ino Y,
Shimosato Y, Yamazaki S, Makuuchi M, Hasegawa
H, Terada M, Hosoda Y: Increasing incidence ofhepatocellular carcinoma possibly associated with
Non-A, Non-B hepatitis in Japan, disclosed by hepatitis B virus DNA analysis of surgically resected cases.
Cancer Research 48:7294-7297, 1989.
12. Makuuchi M, Takayama T, Gunven P, Kosuge T,
Yamazaki S, Hasegawa H: Restrictive versus liberal
blood transfusion policy for hepatectomies in cirrhotic patients. World Journal of Surgery 13:644-648,
1989.
13. Makuuchi M, Thai B.L, Takayasu K, Takayama T,
Kosuge T, Gunven P, Yamazaki S, Hasegawa H,
Ozaki H: Preoperative portal embolization to increasesafety of major hepatectomy for hilar bile duct carcinoma: a preliminary report. Surgery 107:521-527,
1990.
14. Takayama T, Makuuchi M, Hirohashi S, Sakamoto
M, Okazaki N, Takayasu K, Kosuge T, Motoo Y,
Yamazaki S, Hasegawa H: Malignant transformationof adenomatous hyperplasia to hepatocellular carcinoma. Lancet 33:1153-1155, 1990.
15. Kitahara S, Makuuchi M, Kawasaki S, Ishizone S,
Matsunami H, Kamada N, Kawarazaki H, Imanaka T:
Lyphoproliferative disorders after FK 506. Lancet
337:1234, 1991.
16. Kawasaki S, Makuuchi M, Ishizone S, Matsunami H,
Terada M, Kawarazaki H: Liver regeneration in recipients and donors after transplantation. Lancet
339:580-581, 1992.
17. Takayasu K, Wakao F, Moriyama N, Muramatsu Y,
Yamazaki S, Kosuge T, Takayama T, Okada S,
Okazaki N, Makuuchi M: Postresection recurrence ofhepatocellular carcinoma treated by arterial
embolization: Analysis of prognostic factors.
Hepatology 16:906-911, 1992.
18. Matsunami H, Makuuchi M, Kawasaki S, Ishizone S,
Mizusawa Y, Kawarazaki H, Takayama T: Livingrelated liver transplantation in fulminant hepatic failure. Lancet 340:1411-1412, 1992.
19. Makuuchi M, Kawasaki S, Noguchi T, Hashikura Y,
Matsunami H, Hayashi K, Harada H, Kakazu T,

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Takayama T, Kawarazaki H: Donor hepatectomy forliving related partial liver transplantation. Surgery
113:395-402, 1993.
20. Makuuchi M, Hashikura Y, Kawasaki S, Tan D,
Kosuge T, Takayama T: Personal experience of rightanterior segmentectomy (segments V and VIII. forhepatic malignancies. Surgery 114:52-581993.
21. Kawasaki S, Makuuchi M, Matsunami H, Hashikura
Y, Ikegami T, Chisuwa H, Ikeno T, Noike T,
Takayama T, Kawarazaki H: Preoperative measurement of segmental liver volume of donors for living
related liver transplantation. Hepatology 18:11151120, 1993.
22. Hashikura Y, Makuuchi M, Kawasaki S, Matsunami
H, Ikegami T, Nakazawa Y, Kiyosawa K, Ichida T:
Successful living-related partial liver transplantation
to an adult patient. Lancet 343:1233-1234, 1994.
23. Matsushita K, Kawasaki S, Makuuchi M: Arterial
ketone body ratio in liver surgery. Hepatology
20:331-335, 1994.
24. Hui A-M, Kawasaki S, Makuuchi M, Nakayama J,
Ikegami T, Miyagawa S: Liver injury following
normethermic ischemia in steatotic rat liver.
Hepatology 20:1287-1293, 1994.
25. Urata K, Kawasaki S, Matsunami H, Hashikura Y,
Ikegami T, Ishizone S, Momose Y, Komiyama A,
Makuuchi M: Calculation of child and adult standard
liver volume for liver transplantation. Hepatology
21:1317-1321, 1995.
26. Ichida T, Matsunami H, Kawasaki S, Makuuchi M,
Harada T, Itoh S, Asakura H: Living related-donorliver transplantation from adult to adult for primarybiliary cirrhosis. Annals of Internal Medicine
122:275-276, 1995.
27. Miyagawa S, Kawasaki S, Makuuchi M: Carcinoma
between hepatitis B and C viral infection : Tumor
multicentricity in cirrhotic liver with hepatic C.
Hepatology 24:307-310, 1996.
28. Nagai M, Kubota K, Kawasaki S, Takayama T, BandaiY, Makuuchi M: Are left-sided gallbladders reallylocated on the left side? Annals of Surgery 225:274280, 1997.
29. Kubota K, Makuuchi M, Kusaka K, Kobayashi T,
Miki K, Hasegawa K, Harihara Y, Takayama T:
Measurement of liver volume and hepatic functional
reserve as a guide to decision-making in resectional
surgery for hepatic tumors. Hepatology 26:11761181, 1997.
30. Kawasaki S, Makuuchi M, Matsunami H, Hashikura
Y, Ikegami T, Nakazawa Y, Chisuwa H, Terada M,
Miyagawa S: Living related liver transplantation in
adults. Annals of Surgery 227:269-274, 1998.
31. Abe H, Yamada N, Kamata K, Kumaki T, Shimada M,
Osuga J, Shionoiri F, Yahagi N, Kadowaki T,
Tamemoto H, Ishibashi S, Yazaki Y, Makuuchi M:
Hypertension, hypertriglyceridemia and impaired
endothelium-dependent vascular relaxation in micelacking insulin receptor substrate-1. Journal of
Clinical Investigation 101:1784-1788, 1998.

238

32. Takayama T, Makuuchi M, Hirohashi S, Sakamoto
M, Yamamoto J, Shimada K, Kosuge T, Okada S,
Takayasu K, Yamazaki S: Early hepatocellular carcinoma as an entity with a high rate of surgical cure.
Hepatology 28(5.
:1241-1246, 1998.
33. Takayama T, Sekine T, Kondo Y, Kakizoe T,
Makuuchi M: Adjuvant adoptive immunotherapy
against hepatocellular carcinoma. Hepatology
28:1436-1437, 1998.
34. Hui AM, Cui X, Makuuchi M, Li X, Shi YZ,
Takayama T: Decreased p27 (Kip1.
expression
and cyclin D1 overexpression, alone and in combination, influence recurrence and survival of patientswith resectable extrahepatic bile duct carcinoma.
Hepatology 30:1167-1173, 1999.
35. Tanaka S, Mori M, Sakamoto Y, Makuuchi M,
Sugimachi K, Wands JR: Biologic significance ofangiopoietion –2 expression in human hepatocellular
carcinoma. Journal of Clinical Investigation
1103:341-345, 1999.
36. Imamura H, Shimada R, Kubota M, Matsuyama Y,
Nakayama A, Miyagawa S, Makuuchi M, Kawasaki
S: Preoperative portal vein embolization : An audit of
84 patients. Hepatology 229:1099-1105, 1999.
37. Torzilli G, Minagawa M, Takayama T, Inoue K, Hui
AM, Kubota K, Ohtomo K, Makuuchi M: Accurate
preoperative evaluation of liver mass lesions withoutfine-needle biopsy. Hepatology 330:889-893, 1999.
38. Kosuge T, Yamamoto J, Shimada K, Yamasaki S,
Makuuchi M: Improved surgical results for hilar
cholangiocarcinoma with procedures including major
hepatic resection. Annals of Surgery 230:663-671,
1999.
39. Sugawara Y, Makuuchi M, Kato N, Shimotohno K,
Takada K: Enhancement of hepatitis C virus replication by Epstein-Barr virus-encoded nuclear antigen 1.
The EMBO (European Molecular Biology
Organization) Journal 18: 5755-5760, 1999.
40. Hui AM, Li X, Shi YZ, Torzilli G, Takayama T,
Makuuchi M p27(kip) Expression in normal epithelia, precancerous lesions, and carcinomas of thegallbladder: association with cancer progression a n dprognosis. Hepatology 31:1068-1072, 2000.
41. Kogure K, Kuwano H, Fujimaki N, Makuuchi M:
Relation among portal segmentation, proper hepaticvein, and external notch of the caudate lobe in thehuman liver. Annals of Surgery 231:223-228, 2000.
42. Minagawa M, Makuuchi M, Torzilli G, Takayama T,
Kawasaki S, Kosuge T, Yamamoto J, Imamura H:
Extension of the frontiers of surgical indications in
the treatment of liver metastases from colorectal cancer. Annals of Surgery 231:487-499, 2000.
43. Miki K, Al-Refaie W, Xu M, Jiang P, Tan Y, Bouret M,
Zhao M, Gupta A, Chishima T, Shimada H,
Makuuchi M, Moossa AR, Hoffman RM:
Methioninase gene therapy of human cancer cells is
synergistic with recombinant methioninase treatment. Cancer Research 60:2696-2702, 2000.
44. Takayama T, Sekine T, Makuuchi M, Yamasaki S,
Kosuge T, Yamamoto J, Shimada K, Sakamoto M,

Hirohashi S, Ohashi Y, Kakizoe T: Adoptive
immunotherapy to lower postsurgical recurrence
rates of hepatocellular carcinoma: a randomised trial.
The Lancet 356(9232. :802-807, 2000.
45. Arii S, Yamaoka Y, Futagawa S, Inoue K, Kobayashi
K, Kojiro M, Makuuchi M, Nakamura Y, Okita K,
Yamada R, For The Liver Cancer Study Group ofJapan: Results of surgical and nonsurgical treatment
for small-sized hepatocellular carcinomas : A
Retrospective and Nationwide Survery in Japan.
Hepatology 32:1224-1229, 2000.
46. Torzilli G, Makuuchi M, Midorikawa Y, Sano K,
Inoue K, Takayama T, Kubota K: Liver resection
without total vascular exclusion hazardous or beneficial? Annals of Surgery 233:167-175, 2001.
47. Minagawa M, Makuuchi M, Takayama T, Ohtomo K:
Selection criteria for hepatectomy in patients with
hepatocellular carcinoma and portal vein tumor
thrombus. Annals of Surgery 233:379-384, 2001.
48. Saiura A, Sato M, HirataY, Nagai R, Makuuchi M: letter to the editor: Cirulating smooth muscle progenitor cells contribute to atherosclerosis. Nature
Medicine 7(4): 382-383, 2001.
49. Kubota K, Takayama T, Sano K, Hasegawa K, Aoki T,
Sugawara Y, Makuuchi M: Small bile duct reconstruction of the caudate lobe in living-related livertransplantation. Annals of Surgery 235(2):174-177,
2002.
50. Sano K, Makuuchi M, Miki K, Maema A, Sugawara
Y, Imamura H, Matsunami H, Takayama T:
Evaluation of hepatic venous congestion proposed
indication criteria for hepatic vein reconstruction.
Annals of Surgery 236:241-247, 2002.

239

Department of Urology
Outline and Research Objectives
Originally, our department had been attached to Department of Dermatology since 1890,
later in 1926, the current department of urology was founded. Initially, urology dealt withurinary tract infection, especially gonorrheal urethritis. Subsequently, its scope has been
expanded dramatically and now is covering the diseases of the adrenal gland, the kidney,
the urinary tract and the male genital system by means of surgical and non-surgical procedures. Its subspecialty includes pediatric urology, neurourology, female urology, renal transplantation, vascular surgery, endocrine surgery and geriatric urology. For this reason, urology in these days is beyond a section of surgery.
During these progresses, we have been taking national and international leadership indeveloping and applying the new or minimally invasive treatment modalities. They are
exemplified by endoscopic management of the diseases in the upper urinary tract, extracorporeal shock wave lithotripsy (ESWL), laser therapies for prostatic hypertrophy and laparoscopic adrenalectomy or nephrectomy substituting open procedures. Nowadays we are
expanding the frontier by introducing less invasive surgeries or more sophisticated treatment modalities to urology. This expansion is supported by incorporating updated cellularand molecular biology to clinical medicine.

Faculties and Students
Professor and Chair

Tadaichi Kitamura, M. D., Ph. D.
(1998-)
Associate Professors Yukio Homma, M. D., Ph. D.,
Nobutaka Ohta, M. D., Ph. D.
Lecturers
Takumi Takeuchi, M. D., Ph. D.,
Satoru Takahashi, M. D., Ph. D.
Kyoichi Tomita, M. D., Ph. D.,
Makoto Suzuki, M. D., Ph. D.
Associate ...............................12
Graduate student ..................11
Research student.....................3
Secretary .................................5

Past Research and Major Accomplishments
1) Clinical accomplishment
We have been applying endoscopic procedures toclinical practice as less invasive treatment modalities.
For example, we are the pioneers of pyeolureteroscopy, by which the upper urinary tract diseases are endoscopically managed.
We have developed new prognostic predictors
such as fluorescence in-situ hybridization analysis of
exfoliated urothelial cells for bladder cancer recurrence. We examined accuracy of diagnostic procedures for voiding dysfunctions and detected anatomical and electrophysiological abnormalities of the striated urethral sphincter in stress urinary incontinent

240

women.
As the new treatment modalities, dendritic cell
immunotherapy was successfully performed inpatients with metastatic renal cell carcinoma.
Magnetic stimulation of the sacral roots was developed to improve urinary incontinence.
2) Basic research
In oncology field, we found that hepatocyte
growth factor, IL4, Fas ligand or caspase-1 enhanced
the invasiveness of renal cell carcinoma, and that antiapoptotic Bcl-2 proto-oncogene expression increased
resistance of human prostatic cancer cells to various
apoptotic stimuli. For bladder cancer we detected frequent agreement of genetic alternations (loss of heterozygosity at 10 microsatellite loci and methylationof p16INK4 CpG-island) in cancerous mucosa and “normal” bladder mucosa, suggesting morphologically normal epithelium possesses genetic or epigenetic aberrations common with cancer.
In nephrology we produced a knockout mousemodel of autosomal dominant polycystic kidney disease (ADPKD) with a targeted deletion of Pkd1 .
Homozygote embryos developed hydrops, cardiac
conotruncal defects and renal cystogenesis partly
through the E-cadherin-b-catenin-c–MYC pathway
and tyrosine phosphorylation of EGFR and Gab1.
Maternally-administered pioglitazone, a thiazolidinedione compound, improved survival of knockout
embryos, the degree of renal cystogenesis andendothelial dysfunction, suggesting thiazolidine-diones as a new therapeutic for ADPKD.
Other basic investigations include transplantation
research to improve graft survival by expressing IL4
or Fas ligand in the graft, and pharmacologic researches to examine adrenoceptor expression in the prostateor to apply adrenomedullin, a vasorelaxant peptide, toerectile dysfunction and nephron sparing surgery.
3) Investigation in urinary virology
Over these 15 years, we have been investigating
the mode of infection in urinary polyomavirus JC
virus (JCV) that causes progressive multifocal leukoencephalopathy in immuno-compromized patients. It
has long been known since 1970’s that JCV is ubiquitous in the human population and infects children
asymptomatically. However, other characteristics of
JCV had not been elucidated further. From our longrun study for about 15 years, many facts have been
elucidated. JCV persists and replicates in renal tissue
throughout life and is shed frequently into urine afterlate adolescent and the excretion rate exceeds 60%
after around age 50.
The main mode of transmission of JCV is fromparents to children through long term cohabitation.
During the course of study, we have identified 3
major types of JCV, types A, B and C, according totheir DNA sequence variance. A distinct relationshipwas demonstrated between JCV genotypes andhuman races. Type A is predominant in Caucasianand type B predominates in Asian, while mostAfricans have type C, which is verified by phylogenetic tree analysis. Recently Japanese-Americans haveshown to reserve their original or Japanese type of JC
virus, which supports the hypothesis that JCV istransmitted mostly within the family through longterm cohabitation. As of the end of September in
2002, many urine samples have been collected fromacross the world more than 50 different countries. In
a result, JCV can be classified into at least 12 JCV
genotypes that occupy unique domains in the world.
Each of 12 JCV genotypes has a special association
with distinct human population. Those results havebeen published in 20 papers of PNAS, J of Virology,
Virology, J of Infectious Diseases, and so on.

Current Research
We are more and more concentrating on prostatecancer as the target disease. The expression level of
estrogen receptor and its responsive genes are analyzed to further understand molecular mechanisms of
responsiveness of prostate cancer cells to estrogens.
The expression level of these molecules in humanspecimens is investigated for predictability of clinicalprogression of the prostate cancer. Cadherin and its
variant forms are also investigated for its predictabili-

ty of invasion and metastasis of the prostate cancer,
urinary bladder cancer and renal pelvic cancer.
Genetic variant of metabolic activation of estramustine phosphate is analyzed for individual patientswith hormone refractory prostate cancer to minimize
adverse events associated with the agent.
Laparoscopic surgery has been established for adrenalectomy, and now being used for prostatectomy,
nephrectomy and nephroureterectomy. Treatment foradvanced cancer is a challenge in oncology. A new
combination chemotherapy regime including paclitaxel and cis-platinum is attempted for metastatic
prostate and bladder cancers. Tissue oxidative stress,
which is measured by the ratio of reduced-to-total coenzyme Q contents, is now investigated in relation tocarcinogenesis, invasion and metastasis of prostatecancer in rats.
Apart form prostate cancer we are developing
instruments to measure symptoms and quality of life
to assess the efficacy of treatment for voiding dysfunction. As an innovative treatment modality for
benign prostatic hyperplasia, high-intensity-focusedultrasound that evaporates prostatic tissue is being
applied to reduce the urethral obstruction. Alphaadrenoceptor is known to mediate tonus of bladderoutlet. Knock out mice of the receptors are being
investigated for their voiding habits and physiologicalcharacteristics of the prostate and bladder. We are
exploring genetic abnormality in recurrent stone formers with special interest in genes related to sodium
dicarboxylate cotransporter, which transports citrate
in renal tubules. For virology we are now examining
the offspring of mixed marriages to address the issueof whether JCV has a paternal or maternal bias to its
transmission to children. JC virus genotypes in AINU
people and Antarctic people are analyzed to obtainfurther information on migration of human-beings in
the history..

Future Prospect
Of clinical relevance, improving skills for endoscopic surgeries, especially for laparoscopic procedures
are mandatory.
In the field of basic research, tailor-made medicine
for advanced prostate cancer will be mainly pursued
hereafter, as well as elucidating the pathogenesis ofurological malignancies. Besides oncology, clinical andbasic research will be performed in the fields of neurogenic bladder, female urology, geriatric urology,
urolithiasis, erectile dysfunction and urinary virology.

Research Grants
1. Grant source Grant-in-Aid for Scientific Research (A)from the Ministry of Education, Science, Sports and

241

2.

3.

4.

5.

Culture, Japan Research title Application of genetherapy in urologic oncology Principle investigator
Tadaichi Kitamura Fund amount 28,800,000 Yen
Term 1999-2002
Grant source Grant-in-Aid for Scientific Research (C)from the Ministry of Education, Science, Sports andCulture, Japan Research title Prevention of prostate
carcinogenesis by suppressing tissue oxidative stressPrinciple investigator Yukio Homma Fund amount
3,200,000 Yen Term 2001-2004
Grant source Grant-in-Aid for Scientific Research (B)from the Ministry of Education, Science, Sports andCulture, Japan Research title Functional role ofadrenoceptor subtype in lower urinary tract obstruction in knockout mice Principle investigator
Nobutaka Ohta Fund amount 14,700,000 Yen Term
2001-2002
Grant source Grant-in-Aid for Scientific Research (C)from the Ministry of Education, Science, Sports andCulture, Japan Research title Estrogen receptors andestrogen receptor responsive genes in prostate cancer
and benign hyperplasia Principle investigator Satoru
Takahashi Fund amount 4,000,000 Yen Term 20022003
Grant source Grant-in-Aid for Scientific Research (C)from the Ministry of Education, Science, Sports andCulture, Japan Research title Role of cadherins ininvasion and metastasis of prostate cancer Principle
investigator Kyoichi Tomita Fund amount 3,600,000
Yen Term 2001-2002

Select Publications
1. Guo J, Sugimoto C, Kitamura T, Ebihara H, Kato A,
Guo Z, Liu J, Zheng SP, Wang YL, Na YQ, Suzuki M,
Taguchi F and Yogo Y. Four geographically distinct
genotypes of JC virus are prevalent in China andMongolia: implications for the racial composition ofmodern China. J. Gen. Virol. 79, 2499-2505, 1998.
2. Hara K, Sugimoto C, Kitamura T, Aoki N, Taguchi F
and Yogo Y. Archetype JC virus efficiently replicatesin COS-7 Cells, simian cells constitutively expressing
simian virus 40 T antigen. J. Virol. 72, 5335-5342,
1998.
3. Homma Y, Gotoh M, Takei M, Kawabe K and
Yamaguchi T. Predictability of conventional tests for
bladder outlet obstruction assessment in benign prostatic hyperplasia. Int. J. Urol. 5, 61-66, 1998.
4. Homma Y, Kawabe K and Hayashi K. Urological morbidity and its influence on global satisfaction withtreatment outocome after radical prostatectomy forprostate cancer. Int. J. Urol. 5, 556-561, 1998.
5. Kitamura T, Sugimoto C, Ebihara H, Kato A, Guo J,
Taguchi F, Tominaga T, Ogawa Y, Ohta N, Kizu N,
Imamura K, Funaki H, Kurosawa T, Ichikawa S,
Suzuki T, Chiba K, Nagashima K, Yasumoto S andYogo Y. Peopling of Japan as revealed by genotypingof urinary JC virus DNA. Anthropol. Sci. 4,311-325,
1998.

242

6. Aoki N, Kitamura T, Tominaga, T, Fukumori N,
Sakamoto Y, Kato K and Mori M.
Immunohistochemical detection of JC virus in
Nontumorous renal tissue of a patient with renal
cancer but without progressive multifocal leukoencephalopathy. J. Clin. Microbiol. 37,1165-1167, 1999.
7. Homma Y, Akaza H, Okada K, Yokoyama M,
Moriyama N, Usami M, Hirao Y, Tsushima T,
Sakamoto A, Ohashi Y, Aso Y and Prostate Cancer
Study Group. Early results of radical prostatectomy
and adjuvant endocrine therapy for prostate cancer
with or without preoperative androgen deprivation.
Int. J. Urol. 6, 229-237,1999.
8. Horie S, Aruga S, Kawamata H, Okui N, Kakizoe T
and Kitamura T. Biological role of HGF/MET pathway in renal cell carcinoma. J. Urol. 161, 990-997,
1999.
9. Kajiwara T, TakeuchiI T, Ueki T, Moriyama N, Ueki
K, Kakizoe T and Kawabe K. Effect of Bcl-2 overexpression in human prostate cancer cells in vitro andin vivo. Int. J. Urol. 6, 520-525, 1999.
10. Nishimatsu H, Takeuchi T, Ueki T, Kajiwara T,
Moriyama N, Ishida T, Li B, Kakizoe T and Kitamura
T. Fas ligand expression enhances growth of murine
renal cell carcinoma in vivo. Cancer Immunol.
Immunother. 48, 56-61, 1999.
11. Nishimatsu H, Moriyama N, Hamada K, Ukai Y,
Yamazaki S, Kameyama S, Konno N, Ishida Y, Ishii
Y, Murayama T and Kitamura T. Contractile responses to alpha1- adrenoceptor agonists in isolated
human male and female urethra. BJU Int. 84, 515520, 1999.
12. Suzuki M, Hosaka Y, Matsushima H, Goto T,
Kitamura T and Kawabe K. Butyro- lactone I induced
cyclin B1, G2/M arrest, and skipping of mitosis in
human prostatic cell Lines. Cancer Lett. 57, 121-130,
1999.
13. Takeuchi T, Ueki T, Nishimatsu H, Kajiwara T,
Ishida T, Jishage K, Ueda O, Suzuki H, Li B,
Moriyama N and Kitamura T. Accelerated rejectionof Fas-ligand expressing heart grafts. J. Immunol.
162, 518-522, 1999.
14. Aruga S, Wehrli S, Kaissling B, Moe O W, Preisig P A,
Pajor A M, and Alpern R J. Chronic metabolic acidosis increases NaDC-1 mRNA and protein abundance
in rat kidney. Kidney Int. 58, 206-215, 2000.
15. Fujishiro T, Enomoto H, Ugawa Y, Takahashi S,
Ueno S and Kitamura T. Magnetic stimulation of thesacral roots in the treatment of stress incontinence:an investigational study and a placebo controlled
trial. J.Urol. 164, 1277-1279, 2000.
16. Homma Y, Yamaguchi O, Kageyama S, Nishizawa O,
Yoshida M and Kawabe K. Nocturia in the adult:
Classification on the basis of the largest voided volume and nocturnal urine production. J.Urol. 163,
777-781, 2000.
17. Homma Y, Hamade K, Nakayama Y, Tsujimoto G
and Kawabe K. Effects of castration on contraction
and alfa1-adrenoceptor expression in rat prostate.
Brit. J. Pharmacol. 131, 1454-1460, 2000.

18. Homma Y, Kondo Y, Takahashi S, Kitamura T and
Kawabe K. Reproducibility of cystometry in overactive detrusor. Eur. Urol. 38, 681-685, 2000.
19. Kato A, Sugimoto C, Zheng HY, Kitamura T and Yogo
Y. Lack of disease-specific amino acid changes in theviral proteins of JC virus isolates from the brain with
progressive multifocal leukoencephalopathy. Arch.
Virol. 145, 2173-2182, 2000.
20. Kume H, Oda H, Nakatsuru Y, Shimizu Y, Tkahashi
Y, Matsumoto T, Inoue S, Kameyama S, Kitamura T
and Ishikawa T. Genetic idenitification of bilateral
primary or metastatic nonpapillary renal cell carcinoma. BJU Int. 86, 208-212, 2000.
21. Muto S, Horie S, Takahashi S, Tomita K and
Kitamura T. Genetic and epigenetic alteration in normal bladder epithelium in patients with metachronous bladder cancer. Cancer Res. 60, 4021-4025,
2000.
22. Takahashi S, Homma Y, Fujishiro T, Hosaka Y,
Kitamura T and Kawabe K. Electromyographic studyof the striated urethral sphincter in type 3 stressincontinence: evidence of myogenic-dominant damages. Urology 56, 946-950, 2000.
23. Takeuchi T, Nishimatsu H, Ueki T, Kajiwara T,
Fukuhara H, Ishida T, Moriyama N and Kitamura T.
Differentially expressed mRNAs in androgen-independent but not androgen-dependent Shionogi carcinoma. Urol. Res. 28, 82-85, 2000.
24. Tomita K, van Bokhoven A, van Leenders GJ, Ruijter
ET, Jansen CF, Bussemakers MJ, Schalken JA.
Cadherin switching in human prostate cancer progression. Cancer Res. 60, 3650-3654, 2000
25. Tomita K, van Bokhoven A, Jansen CF, Bussemakers
MJ and Schalken JA. Coordinate recruitment of Ecadherin and ALCAM to cell-cell contacts by alphacatenin. Biochem Biophys. Res. Commun. 267, 870874, 2000.
26. Akiyama H, Kurosu T, Sakashita C, Inoue T, Mori S,
Ohashi K, Tanikawa S, Sakamaki H, Onozawa Y,
Chen Q, Zheng H and Kitamura K. Adenovirus is a
key pathogen in hemorrhagic cystitis associated withbone marrow transplantation. Clin. Infect. Dis. 32,
1325-1330, 2001.
27. Fukuhara H, Maruyama T, Nomura S, Oshimura M,
Kitamura T, Sekiya T and Murakami Y. Functional
evidence for the presence of tumor suppressor geneon chromosome 10p15 in human prostate cancers.
Oncogene 20, 314-319, 2001.
28. Fujimura T, Takahashi S, Urano T, Ogawa S, Ouchi
Y, Kitamura T, Muramatsu M and Inoue S.
Differential expression of estrogen receptor beta
(ERbeta) and its C-terminal truncated splice variant
ER betacx as prognostic predictors in human prostatic cancer. Biochem. Biophys. Res. Commun. 289, 692695, 2001.
29. Guo Z, Zheng SP, Sugimoto C, Wang YL, Zheng HY,
Takasaka T, Kitamura T, Guo J and Yogo Y. JC virus
genotypes in Northwestern China: Implications for
its population history. Anthropol. Sci. 3, 203-212,
2001.

30. Ishiwata S, Takahashi S, Homma Y, Tanaka Y,
Kameyama S, Hosaka Y and Kitamura T.
Noninvasive detection and prediction of bladder cancer by fluorescence in situ hybridization analysis of
exfoliated urothelial cells in voided urine. Urology
57, 811-815, 2001.
31. Kamijo T, Sato T, Nagatomi Y and Kitamura T.
Induction of apoptosis by cyclooxygenase-2
inhibitors in prostate cancer cell lines. Int. J. Urol. 8,35-39, 2001.
32. Kanda H, Kubo K, Hamasaki K, Kanda Y, Nakao A,
Kitamura T, Fujita T, Yamamoto K and Mimura T.
Influence of various hemodialysis membranes on theplasma (1-3)-beta-D-glucan level. Kidney Int. 60, 319323, 2001.
33. Kuramochi M, Fukuhara H, Nobukuni T, Kanbe T,
Maruyama T, Ghosh Hara P, Pletcher M, Isomura M,
Onizuka M, Kitamura T, Sekiya T, Reeves RH and
Murakami Y. TSLC1 is tumor-suppressor gene inhuman non-small-cell lung cancer. Nature Genetics
27, 427-430, 2001.
34. Kondo Y, Homma Y, Takahashi S, Kitamura T and
Kawabe K. Transvaginal ultrasonography of urethral
sphincter in continent and incontinent women. J.
Urol. 165, 149-152, 2001.
35. Nishimatsu H, Suzuki E, Nagata D, Moriyama N,
Satonaka H, Walsh K, Sata M, Kangawa K, Matsuo
H, Goto A, Kitamura T and Hirata Y.
Adrenomedullin induces endothelium- dependentvasorelaxation via the phosphatidylinositol 3kinase/akt-dependent pathway in rat aorta. Circul.
Res. 89, 63-70, 2001.
36. Nishimatsu H, Hirata Y, Hayakawa H, Nagata D,
Satonaka H, Suzuki E, Horie S, Takeuchi T, Ohta N,
Homma Y, Minowada S, Nagai R, Kawabe K and
Kitamura T. Effects of intracavernous administrationof adrenomedullin on erectile function in rats.
Peptides 22, 1913- 1918, 2001.
37. Ueki T, Takeuchi T, Nishimatsu H, Kajiwara T,
Moriyama N, Narita Y, Ueki K and Kitamura T.
Silencing of caspase-1 gene occurs in murine andhuman renal cancer cells and confers solid tumor
growth in vivo. Int. J. Cancer 91, 673-679, 2001.
38. Yanai Y, Horie S, Yamamoto K, Yamauchi H, Ikegami
H, Kurimoto M and Kitamura T. Characterization of
the anti-tumor activities of IFN-alfa8 in renal cell carcinoma cells. J. Interferon Cytokine Res. 21, 11291136, 2001.
39. Azuma T, Horie S, Tomita K, Takahashi T, Tanaka Y,
Kashiwase K, Nieda M, Takeuchi T, Ohta N, Shibata
Y, Hirai H and Kitamura T. Dendritic cell
immunotherapy for patients with metastatic renal
cell carcinoma: University of Tokyo experience. Int.
J. Urol. 9, 340-346, 2002.
40. Fujishiro T, Takahashi S, Enomoto H, Ugawa Y,
Ueno S and Kitamura T. Magnetic stimulation of thesacral roots in the treatment of urinary frequency
and urge incontinence: an investigational study anda placebo controlled trial. J. Urol. 168, 1036-1039,
2002.

243

41. Homma Y, Yamaguchi T, Kondo Y, Horie S,
Takahashi S and Kitamura T. Significance of nocturia
in the international prostate symptom score forbenign prostatic hyperplasia. J. Urol. 167, 172-176,
2002.
42. Homma Y, Ando T, Yoshida M, Kageyama S, Takei
M, Kimoto K, Ishizuka O, Gotoh M and Hashimoto
T. Voiding and incontinence frequencies - variabilityof diary data and required diary length. Neurourol.
Urodynam. 21, 204-209, 2002.
43. Kitamura T, Nishimatsu H, Hamamoto T, Tomita K,
Takeuchi T and Ohta N. EMP combination
chemotherapy and low-dose monotherapy inadvanced prostate cancer. Expert Rev. Anticancer
Ther. 2, 59-71, 2002.
44. Matsushima H, Kawabe K, Fujime M, Kitamura T,
Homma Y, Kishi H, Kawamura T, Umeda, T, Ohishi,Y, Murai M, Kawai T, Yoshida H and Fukuda T.
Treatment of patients with superficial bladder cancer
by intravesical instillation of anticancer drugs plus
oral chemotherapy following TUR- Bt: A randomized
controlled trial. Oncol. Rep. 9, 283-288, 2002.
45. Muto S, Aiba A, Saito Y, Nakao K, Nakamura K,
Tomita K, Kitamura T, Kurabayashi M, Nagai R,
Higashihara E, Harris PC, Katsuki M and Horie S.
Pioglitazone improves the phenotype and molecular
defects of a targeted Pkd1 mutant. Hum. Mol. Genet.
15, 1731- 1742, 2002.
46. Nishimatsu H, Hirata Y, Shindo T, Kurihara H,
Kakoki M, Nagata D, Hayakawa H, Satonaka H, Sata
M, Tojo A, Suzuki E, Kangawa K, Matsuo H,
Kitamura T and Nagai R. Role of endogenous
adrenomedullin in the regulation of vascular tone
and ischemic renal injury: studies ontransgenic/knockout mice of adrenomedullin gene.
Circul. Res. 90, 657-663, 2002.
47. Saruwatari L, Sugimoto C, Kitamura T, Ohno N,
Sakai E, Shresta P, Hoa BK, Phi PTP, An HPH, Tuyet
NTA, Honjo T, Kobayashi N, Zheng HY, Takasaka T
and Yogo Y. Asian domains of four major genotypesof JC virus, Af2, B1-b, CY and SC. Arch. Virol. 147, 110, 2002.
48. Sugimoto C, Hasegawa, M, Kato A, Zheng HY,
Ebihara H, Taguchi F, Kitamura T, Yogo Y. Evolutionof human polyomavirus JC: Implications for the population history of humans. J.Mol.Evol. 54, 285-297,
2002.
49. Suzuki M, Zheng H-Y, Takasaka T, Sugimoto C,
Kitamura T, Beutler E and Yogo Y. Asian genotypesof JC virus in Japanese-Americans suggests familialtransmission. J. Virol. 2002, in press.
50. Nishimatsu H, Hirata Y, Shindo T, Kurihara H,
Suzuki E, Sata M, Satonaka H, Tanaka R, Nagata D,
Kakoki M, Hayakawa H and Nagai R. Epitherial
responses of the aorta from adrenomedullin transgenic mice and knocckout mice. Hypertens. Res.
2002, in press.

244

Department ofSurgical Oncology
Outline and Research Objectives
The Department of Surgical Oncology was taken over from the First Department ofSurgery, which was established in 1893. Dr. Tetsuichiro Muto, who was the ninth Professorof the First Department of Surgery, was appointed as the first Professor of the Departmentof Surgical Oncology in 1997 based on the introduction of a new system for postgraduate
education. Dr. Hirokazu Nagawa was appointed as the second Professor of this department
in 1999.
The main research objective in this department is to augment quality of life in patientswith malignant disorders by applying knowledge of radiology, molecular biology, cell biology and immunology to clinical setting as well as surgical techniques.
The Department of Surgical Oncology provides comprehensive evaluation, diagnosis,
treatment and management for adult patients with both general and oncologic surgicalproblems, in the ambulatory as well as inpatient setting. Additionally, surgical specialities inthe department include the treatment of benign and malignant disorders of the breast andmanagement of malignancies of the gastrointestinal tract (esophageal, gastric, and colorectal). The department is also well known for its innovative therapy for inflammatory bowel
disease. Department specialists have expertise in biological cancer immunotherapy,
chemotherapy for a variety of malignancies, and radiotherapy for rectal cancer. The department was responsible for 309 inpatients and 18,845 outpatients in the 2000 fiscal year, and402 inpatients and 16,663 outpatients in the 2001 fiscal year.

Faculties and Students
Professor and Chair

Hirokazu NAGAWA, M.D., Ph.D.
(1999-)
Associate Professors Toshiaki WATANABE, M.D., Ph.D.
Lecturer
Joji KITAYAMA, M.D., Ph.D.
Associate .................................9
Postdoctoral Fellow ................5
Graduate Students ................16
Research Students...................7
Secretaries ...............................5

Past Research and Major Accomplishments
1. Preoperative radiotherapy for lower rectal cancer
Preoperative radiation therapy for advanced rectal
cancer has been reported to reduce local recurrence instudies in Western countries, but its effectiveness has
not yet found wide acceptance in Japan. The reason
for this seems to be that although the local recurrence
rate is lower in the irradiated group than that in the
non-irradiated group in studies in Western countries,
the rate shows values around 10% (7.8% to 12.4 %),which is almost equal to or somewhat higher than therate after surgical treatment alone in Japan. Thismight be due to the difference between Westerncountries and Japan in terms of surgical procedure inthat the importance of regional lymphadenectomy
including lateral node dissection has been emphasized
for advanced lower rectal cancer in Japan. Thus,
extended lymphadenectomy including lateral nodedissection (EXT-L) contributes to a low incidence of
local recurrence of lower rectal cancer. However, EXTL is frequently associated with impairment of sexual
and urinary functions. We therefore compared theeffectiveness of preoperative radiotherapy with thatof EXT-L in a prospective, randomized, controlled
study as well as a retrospective analysis. Our studies
have suggested that lateral node dissection is not necessary in terms of curability for patients withadvanced carcinoma of the lower rectum who undergo preoperative radiotherapy (reprint numbers 1 and
2).
Thus, preoperative radiotherapy reduces the rateof local recurrence and improves the chance of survival in patients with resectable, advanced rectal carcinoma. However, because not all tumors respond similarly to radiation, sorting out suitable patients isrequired to irradiate tumors rationally. Therefore, weinvestigated the predicting values of radiosensitivity
in human rectal carcinoma. Specifically, p53,
p21/WAF1 and Ku protein expressions in preradiation245

biopsy specimens from patients with primary rectal
carcinoma were analyzed immunohistochemically.
The proteins of p53 and p21 are those related to cell
cycle and apoptosis pathways, and Ku protein is related to the repair of damaged DNA. Our studies haveindicated that immunohistochemistry detection of
p53, p21 and Ku expressions may be useful parameters for selecting patients with rectal carcinoma forpreoperative radiotherapy (reprint numbers 3 and 4).
2. Appropriate application of chemotherapy topatients with colon carcinoma
A combined administration of 5-fluorouracil (5-FU)
and leucovorin is regarded as a standard chemotherapy for patients with colon carcinoma. Not all tumors,
however, respond similarly to the regimen of
chemotherapy. Therefore, we investigated the predicting values of chemosensitivity in patients with colon
carcinoma from viewpoints of metabolism and apoptotic effects of 5-FU on the tumors. Specifically, weinvestigated the effects of thymidylate synthase (TS)and dihydropyrimidine dehydrogenase (DPD) frommetabolic aspects and Bcl-2 family from apoptotic
aspects on the colon carcinoma. Our studies haveindicated that TS is a prognostic factor for colon carcinoma, and that 5-FU adjuvant chemotherapy is appropriate for colon carcinoma with high expression of TS
and low expression of DPD (reprint number 5). Ourstudies have also suggested that the ratio of Bcl-XL(apoptosis inhibitor) to Bax (apoptosis inducer) is
related to chemosensitivity to 5-FU (reprint number
6).
3. Analysis of cell functions from the viewpoint ofanti-tumor immunity
Immunotherapy using dendritic cells (DCs) is amodality for cancer patients. We have analyzed characteristics of DCs for the purpose of applying effective
DCs to clinical setting. We have demonstrated for the
first time the direct activation of human natural killer(NK) cells by DC-NK cell interaction in vitro, suggesting that DCs may have a central role linking the
innate and adaptive immune responses. Moreover, instimulating NK cell function, peripheral blood DCs
appear to have a different potential from monocytederived DCs. Furthermore, the function of dendritic
cells (DCs), antigen-presenting cells that can initiate
and regulate cellular and humoral responses, is highly
influenced by their level of maturation. Immature DCs
may be harmful in anti-tumor immunotherapy,
because they can induce immunotolerance rather thanimmunostimulation. With respect to this issue, wehave demonstrated that DC culture in an anti-CD40
monoclonal antibody-immobilized plate in medium
supplemented with interferon-gamma has a positive
impact on DC maturation and may be optimal for elic-246

iting an antigen-specific T-cell response without theneed for CD4+ T-helper epitopes (reprint number 7).
Another candidate of effector cells on the antitumor immunity is a subset of natural killer (NK) cells
in the liver, which are enhanced by alpha-glycosylceramides. Alpha-glycosylceramides induce antitumor
immunity in various murine cancer models. Our
observations have strongly suggested the potential
usefulness of alpha-glycosylceramides for
immunotherapy of liver cancer in humans based on
their ability to activate CD3-CD56+ NK cells in theliver (reprint number 8).
4. Experimental analysis of hematogenous andlymphatic metastasis on gastrointestinal cancers
We have made efforts to elucidate the mechanismof hematogenous and lymphatic metastasis on gastrointestinal cancers using a flow adhesion system.
With respect to hematogenous metastasis, cultured
colon cancer cells bound to laminin (LM), but not to
fibronectin or vitronectin under the physiologicalshear condition. Most of the tethered cells did notroll, but arrested immediately and spread within 1030 min on LM under the continuous presence of shear
flow. Our data have suggested that LM can mediatefrom tethering to spreading of colon cancer cells
under the blood flow and plays an essential role inhematogenous metastasis. Our studies have also suggested that E-selectin alone can mediate colon cancer
cell lodgment and subsequent metastasis without thecontribution of integrin molecules and that the different distribution of E-selectin ligands may affect theadhesion behavior of colon cancer cells in flow conditions (reprint number 9).
With respect to lymphatic metastasis, our studies
have indicated high affinity between cancer cells andlymphatic endothelial cells (LEC), and suggested thatlymph node metastasis arises from cancer cells adherent to LEC, which can be augmented by an inflammatory stimulus.
5. Projects on upper gastrointestinal tract
Although our study is mainly focused on colorectal
cancer, we have also investigated and achieved gastricand esophageal cancers in terms of scientific research
as well as surgical techniques. Recently, we identified
X-chromosome-linked inhibitor of apoptosis protein(XIAP) and the gene was designated as hRFI, standingfor human Ring Finger homologous to IAP type.
Northern blot analysis showed that in 70% (14 out of20) of esophageal cancer patients, expression of hRFI
in cancerous regions was two or more times higher
than in the corresponding normal tissues (reprint
number 10). The other major accomplishment in thisfield is listed as follows.

1. p21 Waf1/Cip1 expression is a prognostic marker
in curatively resected esophageal squamous cell
carcinoma, but not p27Kip1, p53, or Rb
2. Local resection with lymphadenectomy for earlygastric cancer
3. Quantative analysis of the cyclin expression inhuman esophageal cancer cell lines
4. Extended lymph node dissection for gastric cancer
cases with N2 lymph node metastasis
5. Microvascular anastomosis for additional bloodflow in reconstruction after intrathoracic
esophageal carcinoma surgery
6. Prognostic significance of non-gastric malignancy
after treatment of early gastric cancer
The relationship of macroscopic shape of superficial esophageal carcinoma to depth of invasion andregional lymph node metastasis.

Current Research
The clinical and academic interests of our department are the upper and lower gastrointestinal tract.
We have begun to apply the techniques used inmolecular and cellular biology to our research basedon the past research and major accomplishment mentioned above. The following are the major themes
under research.
1. Cancer therapy targeting to the tumor vessels
2. Immunotherapy with dendritic cells
3. Modulated chemotherapy in association with genetherapy
4. Genetic analysis of colorectal cancer and adenoma
5. Carcinogenesis in superficial early colorectal cancer
6. Prognostic factor of early colorectal cancer
7. Genetic alterations in synchronous and metachronous multiple colorectal cancers
8. Microsatellite instability and a risk of developing
multiple colorectal cancers
9. Surveillance program following colectomy for colorectal cancer
10. The mechanism of liver metastasis of colorectal
cancer
11. Genetic alterations in ulcerative colitis
12. Association study of candidate genes in ulcerative
colitis13. Carcinogenesis in ulcerative colitis
14. Cancer surveillance in ulcerative colitis
15. Association study of candidate genes in Crohn’sdisease
16. Angiogenesis inhibition in peritoneal metastasis ofgastric cancer

Future Prospects
Research projects described in the past researchand the current research are continuously pursued inour department. More specifically, we are interested
in cancer therapy targeting angiogenesis resulting
from tumor development, immunotherapy with dendritic cells and effective chemotherapy for patientswith cancer. We are also interested in investigating
what mechanism is involved in metastasis, and how
the metastasis is inhibited or suppressed in cancer
patients.
It is considered that the need for appropriate clinical and psychological care for outpatients is increasing
due to the rising number of patients with intractable
malignant diseases. The number of major operations
for elderly patients is also increasing. Taking all thesetrends into account, we need to make greater endeavors in our clinical practice and research in order tomeet the demands of today’s society.

Research Grant
1. Grant-in-Aid for Scientific Research from the
Ministry of Education, Culture, Sports, Science andTechnology of Japan, 1997-2001, ¥112,800, 000
2. Grant-in-Aid for Scientific Research from the
Ministry of Health, Labor and Welfare of Japan,
1997-2001, ¥45,500,000
3. Grant-in-Aid for Scientific Research from the
Ministry of Public Management, Home Affairs, Posts
and Telecommunications of Japan, 1997-2001,¥140,600,000

Select Publications (50 papers)
Colorectum
1. Komuro Y, Watanabe T, Hosoi Y, Matsumoto Y,
Nakagawa K, Tsuno N, Kazama S, Kitayama J, Suzuki
N, Nagawa H. The expression pattern of Ku correlates with tumor radiosensitivity and disease freesurvival in patients with rectal carcinoma. Cancer 95,
1199-1205, 2002.
2. Watanabe T, Tsurita G, Muto T, Sawada T, Sunouchi
K, Higuchi Y, Komuro Y, Kanazawa T, Iijima T,
Miyaki M, Nagawa H. Extended lymphadenectomy
and preoperative radiotherapy for lower rectal cancers. Surgery 132, 27-33, 2002.
3. Nagawa H, Muto T, Sunouchi K, Higuchi Y, Tsurita
G, Watanabe T, Sawada T. Randomized, controlled
trial of lateral node dissection vs. nerve-preserving
resection in patients with rectal cancer after preoperative radiotherapy. Dis Colon Rectum 44, 1274-1280,
2001.
4. Kyo K, Muto T, Nagawa H, Lathrop GM, Nakamura
Y. Association of distinct variants of the intestinal247

mucin gene MUC3A with ulcerative colitis andCrohn’s disease. J Hum Genet 46, 5-20, 2001.
5. Masaki T, Sugiyama M, Atomi Y, Matsuoka H, Abe
N, Watanabe T, Nagawa H, Muto T. The indicationof local excision for T2 rectal carcinomas. Am J Surg
181, 133-7, 2001.
6. Shinozaki M, Watanabe T, Kubota Y, Sawada T,
Nagawa H, Muto T: High proliferative activity is
associated with dysplasia in ulcerative colitis. Dis
Colon Rectum 43(Suppl), S34-S39, 2000.
7. Kyo K, Parkes M, Takei Y, Nishimori H, Nagawa H,
Baba S, Muto T, Lathrop GM, Nakamura Y.
Association of ulcerative colitis with rare VNTR alleles of the human intestinal mucin gene, MUC3.
Human Molecular Genetics 8, 307-311, 1999.
8. Shinozaki M, Muto T, Suzuki K, Matsuda K,
Yokoyama T, Watanabe T, Masaki T, Sato H, Araki
S, Nagawa H. Chronic active disease reflects cancer
risk in ulcerative colitis. Jpn J Cancer Res 90, 10661070, 1999.
9. Fu CG, Tominaga O, Nagawa H, Nita ME, Masaki T,
Ishimaru G, Higuchi Y, Tsuruo T, Muto T. Role ofp53 and p21/WAF1 detection in patient selection for
preoperative radiotherapy in rectal cancer patients.
Dis Colon Rectum 41, 68-74, 1998.
10. Tominaga O, Nita ME, Nagawa H, Muto T. Screening
for APC mutation based on detection of truncated
APC proteins. Dig Dis Sci 43, 306-310, 1998.
11. Muto T, Nagawa H, Watanabe T, Masaki T, Sawada
T. Colorectal carcinogenesis. Dis Colon Rectum40(Suppl), S80-S85, 1997.
12. Tominaga O, Nita ME, Nagawa H, Fujii S, Tsuruo T,
Muto T. Expression of cell cycle regulators in humancolorectal cancer cell lines. Jpn J Cancer Res 88, 855860, 1997.

Chemotherapy
13. Nozawa H, Tsukui H, Nishida K, Yakumaru K,
Nagawa H, Sekikawa T. Dihydropyrimidine dehydrogenase expression in preoperative biopsy and surgically resected specimens of gastric carcinoma.
Cancer Chemother Pharmacol 49, 267-73, 2002.
14. Takenoue T, Nagawa H, Matsuda K, Fujii S, Nita ME,
Hatano K, Kitayama J, Tsuruo T, Muto T. Relation
between thymidylate synthase expression and survival in colon carcinoma, and determination of
appropriate application of 5-fluorouracil by immunohistochemical method. Annals of Surgical Oncology
7, 193-198, 2000.
15. Nita ME, Ono-Nita SK, Tsuno N, Tominaga O,
Takenoue T, Sunami E, Kitayama J, Nakamura Y,
Nagawa H. Bcl-XL antisense sensitizes human coloncancer cell line to 5-fluorouracil. Jpn J Cancer Res 91,
825-832, 2000.
16. Saito S, Tsuno N, Nagawa H, Sunami E, Zhengxi J,
Osada T, Kitayama J, Shibata Y, Tsuruo T, Muto T.
Expression of platelet-derived endothelial cell
growth factor correlates with good prognosis inpatients with colorectal carcinoma. Cancer 88, 42-49,
2000.

248

17. Seto Y, Nagawa H, Mori M, Tsuruo T, Muto T. Effectof 5-fluorouracil on gastrointestinal carcinogenesis
induced by N-methyl-N’-nitro-N-nitrosoguanidine inrats. Dig Dis Sci 44, 75-78, 1999.
18. Nita ME, Nagawa H, Tominaga O, Tsuno N, Fujii S,
Sasaki S, Fu CG, Takenoue T, Tsuruo T, Muto T. 5fluorouracil induces apoptosis in human colon cancer
cell lines with modulation of Bcl-2 family proteins. Br
J Cancer 78, 986-992, 1998.
19. Tsai WS, Nagawa H, Muto T. Differential effects ofpolyunsaturated fatty acids on chemosensitivity of
NIH3T3 cells and its transformants. Int J Cancer 70,
357-361, 1997.

Immunology
20. Osada T, Nagawa H, Takahashi T, Tsuno NH,
Kitayama J, Shibata Y. Dendritic cells cultured inanti-CD40 antibody-immobilized plates elicit a highly
efficient peptide-specific T-cell response. J
Immunother 25, 176-84, 2002.
21. Hasebe H, Sato K, Yanagie H, Takeda Y, Nonaka Y,
Takahashi TA, Eriguchi M, Nagawa H. Bcl-2, Bcl-XL
and c-FLIP (L) potentially regulate the susceptibilityof human peripheral blood monocyte-derived dendritic cells to cell death at different developmental
stages. Biomed Pharmacother 56, 144-51, 2002.
22. Osada T, Nagawa H, Kitayama J, Tsuno NH, Ishihara
S, Takamizawa M, Shibata Y. Peripheral Blood
Dendritic Cells, but Not Monocyte-Derived Dendeitic
Cells, Can Augment Human NK Cell Function.
Cellular Immunology 213, 14-23, 2001.
23. Nakayama H, Kitayama J, Muto T, Nagawa H.
Characterization of intracellular cytokine profile ofCD4(+) T cells in peripheral blood and tumor-draining lymph nodes of patients with gastrointestinal
cancer. Jpn J of Clin Oncology 30, 301-305, 2000.
24. Kitayama J, Hidemura A, Saito H, Nagawa H. Shearstress affects migration behaviour of polymorphonuclear cells arrested on endothelium. Cell Immunology
203, 39-46, 2000.
25. Kitayama J, Ikeda S, Kumagai K, Saito H, Nagawa H.
Alpha-6-beta-1 integrin (VLA-6) mediates leukocytes
tether and arrest on laminin under physiologicalshear flow. Cell Immunology 199, 97-103, 2000.
26. Ishihara S, Nieda M, Kitayama J, Osada T, Yabe T,
Kikuchi A, Koezuka Y, Porcelli SA, Tadokoro K,
Nagawa H, Juji T. Alpha-glycosylceramides enhance
the antitumor cytotoxicity of hepatic lymphocytes
obtained from cancer patients by activating CD3CD56+ NK cells in vitro. J Immunol 165, 1659-1664,
2000.
27. Ishihara S, Nieda M, Kitayama J, Osada T, Yabe T,
Ishikawa Y, Nagawa H, Muto T, Juji T. CD8+NKRP1A+ T cells preferentially accumulate in humanliver. Eur J Immunol 29, 2406-2413, 1999.
Metastasis
28. Nakayama H, Kitayama J, Nagawa H. Rat gastric adenocarcinoma cell line BV9 avidly adheres to lymphat-ic endothelium under lymphatic flow condition. J
Exp Clin Cancer Res 21, 289-94, 2002.
29. Yano H, Kitayama J, Hatano K, Tsuno N, Osada T,
Watanabe T, Tsuruo T, Muto T, Nagawa H.
Clustered cancer cells show a distinct adhesion
behavior from single cell form under physiologicalshear conditions. J Exp Clin Cancer Res 20, 407-12,
2001.
30. Sunami E, Tsuno N, Osada T, Saito S, Kitayama J,
Tomozawa S, Tsuruno T, Shibata Y, Muto T, Nagawa
H. MMP-1 is a prognostic marker for hematogenous
metastasis of colorectal cancer. Oncologist 5, 108114, 2000.
31. Kitayama J, Tsuno N, Sunami E, Osada T, Muto T,
Nagawa H. E-selectin can mediate the arrest type ofadhesion of colon cancer cells under physiologicalshear flow. Eur J Cancer 36, 121-127, 2000.
32. Tomozawa S, Nagawa H, Tsuno N, Hatano K, OsadaT, Kitayama J, Sunami E, Nita ME, Ishihara S, Yano
H, Tsuruo T, Shibata Y, Muto T. Inhibition of
haematogenous metastasis of colon cancer in miceby a selective COX-2 inhibitor, JTE-522. Br J Cancer
81, 1274-1279, 1999.
33. Kitayama J, Nagawa H, Tsuno N, Osada T, Hatano K,
Sunami E, Saito H, Muto T. Laminin mediates tethering and spreading of colon cancer cells in physiological shear flow. Br J Cancer 80, 1927-1934, 1999.
34. Maeda M, Nagawa H, Maeda T, Koike H, Kasai H.
Alcohol consumption enhances liver metastasis incolorectal carcinoma patients. Cancer 83, 1483-1488,
1998.

Molecular biology
35. Sasaki S, Nakamura T, Arakawa H, Mori M,
Watanabe T, Nagawa H, Croce CM. Isolation andcharacterization of a novel gene, hRFI, preferentiallyexpressed in esophageal cancer. Oncogene, 21, 502430, 2002.
36. Uchida H, Shinoura N, Kitayama J, Watanabe T,
Nagawa H, Hamada H. Caspase-8 gene transduction
augments radiation-induced apoptosis in DLD-1
cells. Biochem Biophys Res Commun 292, 347-54,
2002.
37. Kazama S, Ajioka Y, Watanabe H, Watanabe T,
Nagawa H. Not infrequent K-ras mutations in
depressed-type early colorectal carcinomas larger
than 10 mm. Jpn J Cancer Res 93, 178-83, 2002.
38. Kato M, Yano K, Matsuo F, Saito H, Katagiri T,
Kurumizaka H, Yoshimoto M, Kasumi F, Akiyama F,
Sakamoto G, Nagawa H, Nakamura Y, Miki Y.
Identification of Rad51 alteration in patients withbilateral breast cancer. J Hum Genet 45, 133-137,
2000.
39. Umetani N, Sasaki S, Watanabe T, Ishigami H, Ueda
E, Nagawa H. Diagnostic primer sets for microsatellite instability optimized for a minimal amount ofdamaged DNA from colorectal tissue samples.
Annals Surg Oncol 7, 276-280, 2000.

40. Ishiguro T, Nagawa H, Naito M, Tsuruo T. Analysis
of novel metastasis-associated gene TI-227. Jpn J
Cancer Res 91, 390-394, 2000.

Stomach and esophagus
41. Ishiguro T, Ishiguro C, Ishiguro G, Nagawa H.
Midazolam sedation for upper gastrointestinalendoscopy: comparison between the states ofpatients in partial and complete amnesia.
Hepatogastroenterology 49, 438-40, 2002.
42. Nita ME, Nagawa H, Tominaga O, Tsuno N, Hatano
K, Kitayama J, Tsuruo T, Domene CE, Muto T. p21
Waf1/Cip1 expression is a prognostic marker in curatively resected esophageal squamous cell carcinoma,
but not p27Kip1, p53, or Rb. Annals Surg Oncol 6,
481-488, 1999.
43. Seto Y, Nagawa H, Muto Y, Kaizaki S, Kitayama J,
Muto T. Preliminary report on local resection withlymphadenectomy for early gastric cancer. Br J Surg86, 526-528, 1999.
44. Fujii S, Tominaga O, Nagawa H, Tsuno N, Nita ME,
Tsuruo T, Muto T. Quantative analysis of the cyclin
expression in human esophageal cancer cell lines. J
Exp Clin Cancer Res 17, 491-496, 1998.
45. Seto Y, Nagawa H, Muto T. Results of extended
lymph node dissection for gastric cancer cases with
N2 lymph node metastasis. Int Surg 82, 257-261,
1997.
46. Nagawa H, Seto Y, Kobori O, Muto T. An experienceof treatment for patients with thoracic esophageal
cancer invading adjacent organs. Jpn J Clin Oncol 27,
248-250, 1997.
47. Nagawa H, Seto Y, Muto T. Recent advances in gastric cancer. Digestion 58(Suppl 1), 62-64, 1997.
48. Nagawa H, Seto Y, Nakatsuka T, Kaizaki S, Muto T.
Microvascular anastomosis for additional blood flowin reconstruction after intrathoracic esophageal carcinoma surgery. Am J Surg 173, 131-133, 1997.
49. Seto Y, Nagawa H, Muto T. Prognostic significance of
non-gastric malignancy after treatment of early gastric cancer. Br J Surg 84, 418-421, 1997.
50. Seto Y, Nagawa H, Muto T. Impact of lymph nodemetastasis on survival in early gastric cancer. World J
Surg 21, 186-190, 1997.

249

Department of Dermatology
Outline and Research Objectives
Department of Dermatology has been chaired by Kunihiko Tamaki M.D.Ph.D. since
1994. The Department of Dermatology celebrated its 100th anniversary in 1990. Originally it
was founded as the Department of Dermatology and Urology, which also encompassed
venereology. In 1946 the Department of Dermatology was separated from that of Urology.
Department of Dermatology had its branch department in Branch Hospital for about 50
years until these two departments joined last year. A professor, two associate professors,four lecturers and nine associates take part in in-patient and out-patient cares as well as
research and teaching activities. We usually have around 10 residents and 8 students of doctoral PhD. course. Thirty doctors who basically belong to our department are currently out
in affiliated hospitals mainly engaged in clinical works there. Additionally there are four
staff members abroad in the US and involved in advanced research activities in cell biology
and molecular biology as postdoctoral fellow.
We have specialized out-patient clinic in which some of doctors who are in affiliated hospitals also participate and each specialized service reflects its own research field in a diseaseoriented manner.
Of course these specialized groups performing their clinical and reseach activities are notexclusive, and there are increasing communications in our department as well as with otherdepartments. Specialized out-patient clinic include the following; atopic dermatitis, psoriasis, collagen diseases including systemic sclerosis, SLE and dermatomyositis, skin surgery
and laser surgery including malignant melanoma, squamous cell carcinoma and basal cell
carcinoma, and infection especially fungal infection. These clinical groups are divided into
three main research groups;
Immunology and allergology group, 2) collagen disease group and 3) skin surgery group.
Immunology and allergology group engaged in atopic dermatitis and psoriasis clinic andfocuses on Langerhans cell and keratinocyte biology research including cytokine andchemokine. Collagen group engaged in collagen disease clinic and focuses mechanisms TGFβ and collagen. Skin surgery group engaged in skin surgery and laser surgery clinic andfocuses of tumor growth, metastasis, fibrosis and dermatoscopic clinical study. Researchgroups are maintained by associate professor or lecturer. These senior investigators have ten
to fifteen year experience as dermatologist and more that seven year experience of researchin Japan as well as in U.S.A. They have one to three associates and students of Ph.D. course
to extend their research. Professors participate and understand the research activities in thisdepartment through conferences which are usually held once to twice a month by eachresearch group.

Faculties and Students
Professor and Chairman Kunihiko Tamaki M.D. Ph.D.(1994-)
Associate Professors Kanako Kikuchi M.D. Ph.D.
Akihiko Asahina M.D. Ph.D.
LecturerMayumi Komine M.D. Ph.D.
Hironobu Ihn M.D. Ph.D.
Hidehisa Saeki M.D. Ph.D.
Takahiro Watanabe M.D. Ph.D.
Associate .................................9
Graduate student ....................8
Secretary .................................3

250

Past Research and Major Accomplishments
1) Immunology and allergology group:
We established a method to purify
murineLangerhans cells(LC), which comprise from
one to three percentage of epidermal cells, over 95%
purity in each experiment and conducted a series ofexperiments. We found that murine LC produced IL12 which is a key cytokine for Th1 induction and that
IL-12 production was inhibited by GM-CSF andenhanced by TGF-b1. Furthermore we found prionrelated protein was expressed in LC as well as ker-atinocytes. Prion-related protein is maintained on LC
after culture suggesting its participation of LC in
peripheral inoculation of prion disease. With regards
to atopic dermatitis (AD) we found that lesional skinof NC/Nga mice, which exhibit AD like skin lesion,shows enhanced expression of Th2 chemokine
TARC/CCL17 and MDC/ CCL22 and their receptor
CCR4. We extended this to human AD and other skindiseases such as bullous pemphigoid, an autoimmune
blistering disease and mycosis fungoides, cutaneous T
cell lymphoma, all exhibit hypereosinophilia and highserum IgE and found that TARC, MDC and CCR4 are
involved in these diseases.
2) Collagen disease group:
TGF-β has been implicated in the pathogenesis offibrosis. In collaboration with skin surgery group we
showed upregulated expression of TGF-β receptors inlesional skin of fibrosis such as systemic sclerosis andlocalized scleroderma and that upregulated expressionof TGF-β receptors is implicated in fibrosis. Fibroblast
proliferation is also important in fibrosis as well as
wound healing. We demonstrated that fibroblast proliferation is mediated via extracellular signal related
kinase (ERK)-dependent pathway. We also studied theexpression of tissue inhibitor of metalloproteinases(TIMPs) and showed that TIMP2 was upregulated byIL-4 via p-38 mitogen-activated protein kinasedependent pathway. With regards to clinical research
we investigated the prevalence and clinical significance of various autoantibodies in collagen diseases.
We also studied the levels of various cytokines andadhesion molecules and their significance.
3) Skin surgery group:
We investigated progressive and metastatic factors
in malignant melanoma and other skin tumors. We
found dysregulated expression of TGF-β and its receptor in basal cell carcinoma and also found that patternof basal cell keratin 14 expression is a possible marker
for tumor progression in Bowen’s disease. We also
reported TIMP-1 and TIMP-2 differently regulate thegrowth of human melanoma cell lines. With regards
to clinical research we established criteria for the diagnosis of pigmented plantar nevi which is useful to differentiate pigmented nevi from early acral lentiginous
melanoma. We also proposed criteria to differentiate
Hutchingson’s sign seen in malignant melanoma fromsubungual nevus.

Current Research1) Immunology and allergology group:
Using the TGF-β primed LC we are trying to
induce Th1 instead of Th2. We succeeded in inducing

Th1 in vitro and trying to induce Th1 in vivo. We continue to characterize murine LC and found LC express
Toll-like receptor (TLR)-2 3 4 and 9. We are trying to
induce Th1 using these receptors. From the results ofmouse and human study we are trying to establish
K14-TARC transgenic mice to further characterize thesignificance of TARC expression in the epidermis.
With regards to clinical research we are analyzing
gene polymorphism of AD and proriasis such as
Eotaxin, TARC, IL-13 and IL-12. We are also investigating the CTACK/CCL27 expression of AD and psoriasis. We are analyzing the human peripheral blood
dendritic cells (DC) for the expression of cutaneous
lymphocyte associated antigen (CLA), fucosyltransferase VII, CCR4 and CCR10, and their function.
2) Collagen disease group:
We are studying the interaction of transcription
factors Sp 1, Smad 3 and p300/CBP and the significance of these transcriptional factors in TGF-β
induced collagen gene upregulation. PKC is importantin collagen accumulation, thus we are studying theregulation of collagen deposition by PKC especially
PKC a and PKCd The regulation of TGF-β receptors by
various cytokines such as EGF and TNF-α are now
being studied. We found overexpression of aVb5 andVb3 integrin on fibroblasts from fibrotic tissue. The
function of this is under investigation using stabletransfectants of integrins. We also are studying theregulation of tenasin gene expression by IL-13, PDGF
and TGF-β.
3) Skin surgery group:
We are now trying to establish the significance ofsentinel lymph mode biopsy in the clinic and trying
to find the metastatic factors in malignant melanoma.
We are analyzing chemokine and chemokine receptor
expression of primary and metastatic melanoma. We
also are studying the effect of narrow band UVB on
human melanocytes in order to understand theenhanced pigmentation seen in patients receiving narrow band UVB therapy.

Future Prospects
Immunology and allergology group: We will continue the murine LC study to further clarify the characteristics of LC and their participation in skin diseases. We will also study the possibility to use LC fortherapy as well as DC from human peripheral blood.
We will try to find novel gene products using purified
LC and analyze their function. We will establish othertransgenic mice such as K-14- CTACK to further analyze the participation of chemokine in skin diseases.
With regards to clinical research we will further
examine other cytokine and chemokine in skin dis-251eases such as AD and psoriasis and try to use theirantagonist for therapy.
Collagen disease group: To further analyze themolecular mechanisms of TGF-β induced collagengene expression, the cross talk of PI3 kinase andSmad pathway will be determined. Significance ofintegrin expression in fibrosis will be determined.
The mechanisms of EGF and FK506 on the expressionof collagen gene expression will be studied in conjunction with the therapeutic approach.
Skin surgery group: DC therapy for malignant
tumors using DC from peripheral blood and skin resident LC will be investigated. We will also analyze
chemokine / chemokine receptor expression in
growth and metastasis of skin tumors.

Research Grants from
1. The Ministry of Education, Science and Culture(A,2002-2005)“Study on the dendritic cells and chemokine in thepathogenesis of skin diseases”
2. The Health Science Research Grants from the
Ministry of Health, Welface and Labor (2002-)“Study to identify organ specific molecules in thepathogenesis of skin diseases”
3. The Health Science Research Grants from the
Minictry of Health, Welface and Labor (2001-2002)“Basic Research for Atopic Dermatitis using Dendritic
cells as a tool”
4. The Ministry of Education, Science and Culture (B.
1157063 1999-2000)“The study of skin specific T cells in the pathogenesis of skin diseases”
5. The Ministry of Education, Scince and culture(B,11470179,1999-2001)“The study to establish Langerhans cell cell line andits use for treatment”

Select Publications
1. Kakinuma T, Wakugawa M, Nakamura K, Hino H,
Matsushima K, Tamaki K. High level of TARC (thymus and activation-regulated chemokine/CCL17) inblister fluid and sera of patients with bullous pemphigoid. Br J Dermatol, in press.
2. Sugaya M, Nakamura K, Watanabe T, Asahina A,
Yasaka N, Koyama Y, et al: Expression of cellularprion-related protein by murine Langerhans cells andkeratinocytes. Journal of Dermatology Sci. 28:126134,2002
3. Hattori N, Komine M, Yano S, Kaneko T, Hanakawa
Y, Hashimoto K, Tamaki K.: Interferon-g, a strong
suppressor of cell proliferation, induces upregulationof karatin K6, one of the inflammatory- and proliferation-associated keratins. J Invest Dermatol 119:403-10, 2002.
4. Kakinuma T, Nakamura K, Wakugawa M, Mitsui H,
Tada Y, Saeki H, Torii H, Komine M, Asahina A,

252

Tamaki K. Serum macrophage-derived chemokine(MDC) levels are closely related with the diseaseactivity of atopic dermatitis. Clin Exp Immunol, 127:
270-3, 2002.
5. Ihn H, Yamane K, Asano Y, Kubo M, Tamaki K. IL-4
upregulates the tissue inhibitor of metalloproteinase2 in dermal fibroblasts via the p38 mitogen-activatedprotein kinase-dependent pathway. J Immunol 168:
1895-902, 2002.
6. Tamaki K, Nakamura K: The role of lymphocyte inhealthy and eczematous skin. Curr Opinion Allergy
Clin. Immunol 1:455-60,2001
7. Kubo M, Ihn H, Yamane K, Asano Y, Jinnin M,
Tamaki K: Differential expression of transforming
growth factor-beta receptors in squamous cell carcinoma. Br J Dermatol. 145: 840-2, 2001
8. Kubo M, Ihn H, Kuwana M, Yamane K, Yazawa N,
Tamaki K: Prevalence in myositis of antibodies recognizing anti-U3 RNA probably in a novel complex
with 22/25 kD protein and not fibrillarin. Clin Exp
Immunol. 126: 339-44, 2001
9. Yazawa N, Ihn H, Yamane K, Kubo M, Fujimoto M,
Kikuchi K, Soma Y, Tamaki K: Elevated circulating
soluble CD30 levels in patients withpolymyositis/dermatomyositis. Br J Dermatol. 145:
676-8, 2001
10. Hoashi T. Kadono T. Kikuchi K. Etoh T. Tamaki K:
Differential growth regulation in human melanoma
cell lines by TIMP-1 and TIMP-2. Biochem Biophys
Res Commun. 288: 371-9, 2001
11. Komine M. Okinaga M. Takeda F. Nashiro K.
Kikuchi K. Murakami T. Soma Y. Tamaki K: Patternsof basal cell keratin 14 expression in Bowen’s disease: a possible marker for tumour progression. Br J
Dermatol. 145: 223-8, 2001
12. Wakugawa M. Nakamura K. Kakinuma T. Onai N.
Matsushima K. Tamaki K: CC chemokine receptor 4
expression on peripheral blood CD4+ T cells reflects
disease activity of atopic dermatitis. J Invest
Dermatol. 117: 188-96, 2001
13. Tamaki K, Sugaya M, Tada Y, Uasaka N, Uehira M,
Nishimoto H, Nakamura K: Epidermal and dermal
gamma-delta T cells. Chem Immunol 79:43-51,2001
14. Asano Y. Ihn H. Yamane K. Yazawa N. Kubo M.
Fujimoto M. Tamaki K: Clinical significance of surfactant protein D as a serum marker for evaluating
pulmonary fibrosis in patients with systemic sclerosis. Arthritis Rheum. 44: 1363-9, 2001
15. Asahina A. Tada Y. Nakamura K. Tamaki K:
Griseofulvin has a potential to modulate the expression of cell adhesion molecules on leukocytes andvascular endothelial cells. Int Immunopharmacol. 1:
75-83, 2001
16. Kubo M. Ihn H. Yamane K. Tamaki K: Up-regulated
expression of transforming growth factor beta receptors in dermal fibroblasts in skin sections frompatients with localized scleroderma. Arthritis
Rheum. 44: 731-4, 2001
17. Tada Y. Asahina A. Nakamura K. Miyazono K.
Tomura M. Fujiwara H. Tamaki K: Transforming

growth factor-beta up-regulates CD40-engaged IL-12
production of mouse Langerhans cells. Eur J
Immunol. 31: 294-300, 2001
18. Kakinuma T. Nakamura K. Wakugawa M. Mitsui H.
Tada Y. Saeki H. Torii H. Asahina A. Onai N.
Matsushima K. Tamaki K: Thymus and activationregulated chemokine in atopic dermatitis: Serum thymus and activation-regulated chemokine level isclosely related with disease activity. J Allergy Clin
Immunol. 107: 535-41, 2001
19. Yazawa N. Ihn H. Yamane K. Etoh T. Tamaki K: The
successful treatment of prurigo pigmentosa withmacrolide antibiotics. Dermatology. 202: 67-9, 2001
20. Ihn H. Yamane K. Kubo M. Tamaki K: Blockade ofendogenous transforming growth factor beta signaling prevents up-regulated collagen synthesis in scleroderma fibroblasts: association with increased
expression of transforming growth factor beta receptors. Arthritis Rheum. 44: 474-80, 2001
21. Kawabata Y. Ohara K. Hino H. Tamaki K: Two kindsof Hutchinson’s sign, benign and malignant. J Am
Acad Dermatol. 44: 305-7, 2001
22. Wakugawa M. Hayashi K. Nakamura K. Tamaki K:
Evaluation of mite allergen-induced Th1 and Th2
cytokine secretion of peripheral blood mononuclear
cells from atopic dermatitis patients: association
between IL-13 and mite-specific IgE levels. J
Dermatol Sci. 25: 116-26, 2001
23. Asahina A. Tada Y. Nakamura K. Tamaki K:
Colchicine and griseofulvin inhibit VCAM-1 expression on human vascular endothelial cells - evidence
for the association of VCAM-1 expression withmicrotubules. J Dermatol Sci. 25: 1-9, 2001
24. Watanabe T. Nakamura K. Wakugawa M. Kato A.
Nagai Y. Shioda T. Iwamoto A. Tamaki K:
Antibodies to molluscum contagiosum virus in thegeneral population and susceptible patients. Arch
Dermatol. 136: 1518-22, 2000
25. Ihn H. Tamaki K: Increased phosphorylation of transcription factor Sp1 in scleroderma fibroblasts: association with increased expression of the type I collagen gene. Arthritis Rheum. 43: 2240-7, 2000
26. Ihn H. Tamaki K: Oncostatin M stimulates thegrowth of dermal fibroblasts via a mitogen-activatedprotein kinase-dependent pathway. J Immunol. 165:
2149-55, 2000
27. Wakugawa M. Nakamura K. Hino H. Toyama K.
Hattori N. Okochi H. Yamada H. Hirai K. Tamaki K.
Furue M: Elevated levels of eotaxin and interleukin-5
in blister fluid of bullous pemphigoid: correlation
with tissue eosinophilia. Br J Dermatol. 143: 112-6,
2000
28. Nakamura K. Imakado S. Takizawa M. Adachi M.
Sugaya M. Wakugawa M. Asahina A. Tamaki K:
Exacerbation of pustulosis palmaris et plantaris after
topical application of metals accompanied by elevated levels of leukotriene B4 in pustules. J Am Acad
Dermatol. 42: 1021-5, 2000

29. Ihn H. Tamaki K: Competition analysis of the humanalpha2(I) collagen promoter using synthetic oligonucleotides. J Invest Dermatol. 114: 1011-6, 2000
30. Tada Y. Asahina A. Nakamura K. Tomura M.
Fujiwara H. Tamaki K: Granulocyte/macrophagecolony-stimulating factor inhibits IL-12 production ofmouse Langerhans cells. J Immunol. 164: 5113-9,
2000
31. Ihn H. Sato S. Fujimoto M. Igarashi A. Yazawa N.
Kubo M. Kikuchi K. Takehara K. Tamaki K:
Characterization of autoantibodies to endothelial
cells in systemic sclerosis (SSc): association with pulmonary fibrosis. Clin Exp Immunol. 119: 203-9, 2000
32. Yazawa N. Kikuchi K. Ihn H. Fujimoto M. Kubo M.
Tamaki T. Tamaki K: Serum levels of tissue inhibitorof metalloproteinases 2 in patients with systemicsclerosis. J Am Acad Dermatol. 42: 70-5, 2000
33. Yamane K. Ihn H. Kubo M. Yazawa N. Kikuchi K.
Soma Y. Tamaki K: Increased serum levels of solublevascular cell adhesion molecule 1 and E-selectin inpatients with localized scleroderma. J Am Acad
Dermatol. 42: 64-9, 2000
34. Salgado CG, Nakamura K, Sugaya, M, Tada Y,
Asahina A, Fukada, S, Koyama Y, Irie S, Tamaki K.
Differential effects of cytokines and immunosuppressive drugs on CD40, B7-1, and B7-2 expression on
purified epidermal Langerhans cells. J Invest
Dermatol 113: 1021-7, 1999.
35. Kanda N, Tamaki K. Ganglioside GD1b suppresses
immunoglobulin production by human peripheral
blood mononuclear cells. Exp Hematol 27: 1487-93,
1999.
36. Sugaya M, Nakamura K, Tamaki K. Interleukins 18
and 12 synergistically upregulate interferon-g production by murine dendritic epidermal T cells. J
Invest Dermatol 113: 350-4, 1999.
37. Salgado CG, Nakamura K, Sugaya M, Tada Y,
Asahina A, Koyama Y, Irie S, Tamaki K. Functional
CD40 ligand is expressed on epidermal Langerhans
cells. J Leukocyte Biol 66: 281-5, 1999.
38. Kubo M, Ihn H, Sato S, Kikuchi K, Tamaki K.
Prevalence and antigen specificity of anti-histone
antibodies in patients with polymyositis/dermatomyositis. J Invest Dermatol 112: 711-5, 1999.
39. Watanabe T, Tsuchida T, Kanda N, Mori K, Hayashi
Y, Tamaki K. Anti-a-fodrin antibodies in Sjogren syndrome and lupus erythematosus. Arch Dermatol 135:
535-9, 1999.
40. Kanda N, Tamaki K. Estrogen enhances
immunoglobulin production by human PBMCs. J
Allergy Clin Immunol 103: 282-8, 1999.
41. Kanda N, Tsuchida T, Tamak K. Estrogen enhancement of anti-double-stranded DNA antibody andimmunoglobulin G production in peripheral bloodmononuclear cells from patients with systemic lupus
erythematosus. Arthritis Rheum 42: 328-37, 1999.
42. Kubo M, Kikuchi K, Nashiro K, Kakinuma T, Hayashi
N, Nanko H, Tamaki K. Expression of fibrogenic
cytokines in desmoplastic malignant melanoma. Br J
Dermatol 139: 192-7, 1998.

253

43. Kanda N, Tamak K. Ganglioside CQ1b enhances Ig
production by human PBMCs. J Allergy Clin
Immunol 102: 813-20, 1998.
44. Kubo M, Kikuchi K, Fujimoto M, Tamaki K. Serum
concentration of carboxyterminal propeptide of type1 procollagen and tissue inhibitor of metaloproteinase 1 in patients with dermatomyositis. Arch
Dermatol Res 290: 253-7, 1998.
45. Kawabata Y, Tamaki K. Distinctive dermatoscopic
features of acral lentiginous melanoma in situ fromplantar melanocytic nevi and their histopathologic
correlation. J Cutan Med Surg 2: 199-204, 1998.
46. Kikuchi K, Kubo M, Kadono T, Yazawa N, Ihn H,
Tamaki K. Serum concentrations of vascularendothelial growth factor in various collagen diseases. Br J Dermatol 139: 1049-51, 1998.
47. Watanabe T, Tsuchida T, Ito Y, Kanda N, Ueda Y,
Tamaki K. Annular erythema associated with lupus
erythematosus / Sjogren’s syndrome. J Am Acad
Dermatol 36: 214-8, 1997.
48. Kanda N, Tsuchida T, Tamaki K. Testosterone suppresses anti-DNA antibody production in peripheralmononuclear cells from patients with systemic lupus
erythematosus. Arthritis Rheum 40: 1703-11, 1997.
49. Kikuchi K, Kadono T, Furue M, Tamaki K. Tissue
inhibitor of metalloproteinase-1 (TIMP-1) may be an
autocrine growth factor in scleroderma fibroblasts. J
Invest Dermatol 108: 281-4, 1997.
50. Furue M, Kato M, Nakamura K, Nashiro K, Kikuchi
K, Okochi H, Miyazono K Tamaki K. Dysregulated
expression of transforming growth factor β and its
type I and II receptors in basal cell carcinoma. Int J
Cancer 71: 505-9, 1997.

254

Department of Plastic andReconstructive Surgery
Outline and Research Objectives
The Department of Plastic and Reconstructive Surgery was established in 1960 as thefirst independent clinical department in Japan. Clinical activities were initially focused oncongenital anomalies and skin surgery. In the 1970’s, however, surgical techniques such as
microsurgery, craniofacial surgery, tissue expansion, etc. were developed, and our activities
were significantly expanded so that could lead the world in the field of the free tissue transfers, head and neck cancer reconstruction, hand surgery, craniomaxillofacial surgery, etc.
Recently, cosmetic surgery and aesthetic dermatology were also added to our clinical field,
and now we have the greatest number of patients for cosmetic purposes among all universities in Japan. Current research objectives are tissue and cell engineering from somatic stem
cells, hair regeneration, anti-aging effects of hormones on skin, embryonic development ofcongenital anomaly, and application of retinoids to skin and keloids.

Faculties and Students:
Professor and Chair Kiyonori Harii, M.D. (1989-)
Associate Professors Hirotaka Asato, M.D. (1998-)
Lecturer
Kotaro Yoshimura, M.D. (1998-)
Associates ...............................4
Postdoctral Fellows.................5
Graduate Students ..................4
Secretaries ...............................2

Past Research and Major Accomplishments
Professor Harii has developed a number of surgical
techniques in reconstructive microsurgery such asmuscle transplantation for facial reanimation in facialparalyzed patients, and microsurgical reconstructionof head and neck cancer patients. These activities
were published in various international medical journals and textbooks. He accepted a number of trainees
from countries all over the world, who studied andmastered microsurgery in our laboratory.

Current Research
We have several research projects, both basic andclinical. In basic research, molecular and cellularapproaches to reveal the mechanism of congenital
anomalies, such as cleft lips and palates, ear deformities, and craniofacial deformities, are ongoing. We are
seeking genes which can cause those anomalies. In
addition, influences of hormones, especially estrogen,on skin aging are being investigated. Genes, which
can be upregulated or downregulated by estrogen indermal fibroblasts, are being investigated by microarray technique. We are trying to determine the mechanism by which estrogen affects the chronologicalaging of skin. The third one is on hair regeneration.
We harvest and culture dermal papilla cells from hairfollicles, which can transform epidermal stem cells
into hair follicles. We have tried to determine criticalfactors released by dermal papilla cells in order toapply to a practical treatment of alopecia. The last one
is mesenchymal stem cells extraction from lipoaspirates. We are trying to transform the stem cells into
adipocytes, chondrocytes, osteoblasts, dermal fibroblasts, and others for clinical use. The optimal protocols to culture and transform the stem cells have been
investigated.
Clinically, we have attempted to establish a classification of pigmented skin lesions. Also, the mechanism of hyperpigmentation and rational therapies are
currently being established. Clinical effects of achemopreventional therapy for keloids using retinoids
are now under estimation.

Future Prospects
Our research projects will be more focused on tissue and cell engineering using somatic (mesenchymal)
stem cells in the future. Cosmetic augmentation ofsoft tissue such as breasts, cheek, and eyelids may beclinically available using adipose cells or tissues produced by mesenchymal stem cells extracted fromlipoasipates. The stem cells could be applied to therapies of a wide variety of diseases such as hepatic dysfunction, arterial sclerosis, joint cartilage problems,
myocardial infarction, leukemia, etc.

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Research Grants
1) Kotaro Yoshimura, Grants-in aid for scientific research(B) 2002-2003, Molecular biological research on
MMPs and TIMPs expression of human keloids.
2) Hirotaka Asato, Grants-in aid for scientific research (B)2002-2003, Study on a specific skeletogenic potentialof the cranial neural crest cells.
3) Kiyonori Harii, Grants-in aid for scientific research (A),
2000-2001, Evaluation of rejection at the transplantation of cultured cell and induction of local torelance
without systemic immunosuppuression.
4) Kotaro Yoshimura, Grants-in aid for scientific research(B), 2000-2001, Molecular biological research on
actions of retinoids on wound healing and pigmentation in skin.
5) Shinichi Wakita, Grants-in aid for scientific research(B), 1999-2000, Mesenchymal stem cells; actions inwound healing and application to tissue engineering.

Select Publications
1. Takushima A, Asato H, Harii K, Masashi S.
Simultaneous harvest of intercostal nerves and elevation of rectus abdominis musculocutaneous flap for
facial nerve cable grafting. Plast Reconstr Surg 110,541-544, 2002.
2. Yoshimura K, Asato H, Jejurikar SS, Cederna PS,
Urbanchek MG, Kuzon WM Jr. The effect of two
episodes of denervation and reinnervation on skeletal muscle contractile function. Plast Reconstr Surg
109, 212-219, 2002.
3. Takushima A, Harii K, Asato H, Yamada A.
Neurovascular free-muscle transfer to treat facial
paralysis associated with hemifacial microsomia.
Plast Reconstr Surg 109, 1219-1927, 2002.
4. Kimata Y, Uchiyama K, Sakuraba M, Ebihara S,
Hayashi R, Haneda T, Onitsuka T, Asakage T,
Nakatsuka T, Harii K. Velopharyngeal function after
microsurgical reconstruction of lateral and superior
oropharyngeal defects. Laryngoscope 112, 10371042, 2000.
5. Harii K., Asato H.: Facial reanimation. The
Unfavorable Result in Plastic Surgery, 3rd Ed. (ed. by
Goldwyn, R.M., Cohen, M.N.), Lippincott, Williams
and Wilkins, Philadelphia, 2001
6. Kitano Y, Yoshimura K, Uchida G, Sato K, Harii K.
Pretreatment with topical all-trans-retinoic acid isbeneficial for wound healing in genetically diabetic
mice. Arch Dermatol Res 293, 515-521, 2001.
7. Ichioka S, Nakatsuka T, Ohura N, Sato Y, Harii K.
Clinical use of amrinone (a selective phosphodiesterase III inhibitor) in reconstructive surgery. Plast
Reconstr Surg 108, 1931-1937, 2001.
8. Nagase T, Nakamura S, Harii K, Osumi N. Ectopically
localized HNK-1 epitope perturbs migration of the
midbrain neural crest cells in Pax6 mutant rat. Dev
Growth Differ 43, 683-692, 2001.
9. Takushima A, Harii K, Asato H, Nakatsuka T, Kimata
Y. Mandibular reconstruction using microvascular

256

free flaps: a statistical analysis of 178 cases. Plast
Reconstr Surg 108, 1555-1563, 2001.
10. Yoshimura K, Tsukamoto K, Okazaki M, Virador
VM, Lei TC, Suzuki Y, Uchida G, Kitano Y, Harii K.
Effects of all-trans retinoic acid on melanogenesis inpigmented skin equivalents and monolayer culture ofmelanocytes. J Dermatol Sci 27, S68-75, 2001.
11. Nakagawa M, Miyagoe-Suzuki Y, Ikezoe K, Miyata Y,
Nonaka I, Harii K, Takeda S. Schwann cell myelination occurred without basal lamina formation inlaminin alpha2 chain-null mutant (dy3K/dy3K) mice.
Glia 35, 101-110, 2001.
12. Yoshimura K, Harii K, Masuda Y, Takahashi M,
Aoyama T, Iga T. Usefulness of a narrow-bandreflectance spectrophotometer in evaluating effects
of depigmenting treatment. Aesthetic Plast Surg 25,
129-133, 2001.
13. Nagase T, Shimoda Y, Sanai Y, Nakamura S, Harii K,
Osumi N.: Differential expression of two glucuronyltransferases synthesizing HNK-1 carbohydrate epitope in the sublineages of the rat myogenic progenitors. Mech Dev 98, 145-149, 2000,.
14. Kimata Y, Uchiyama K, Ebihara S, Saikawa M,
Hayashi R, Haneda T, Ohyma W, Kishimoto S, Asai
M, Nakatsuka T, Harii K.: Postoperative complications and functional results after total glossectomy
with microvascular reconstruction. Plast Reconstr
Surg 106, 1028-1035, 2000.
15. Kimata Y, Uchiyama K, Ebihara S, Sakuraba M, Iida
H, Nakatsuka T, Harii K. Anterolateral thigh flap
donor-site complications and morbidity. Plast
Reconstr Surg 106, 584-589, 2000.
16. Ichioka S, Nakatsuka T, Ohura N, Sato Y, Harii K.:
Topical application of amrinone (a selective phosphodiesterase III inhibitor) for relief of vasospasm. J SurgRes 93, 149-155, 2000.
17. Gonda K, Nakaoka T, Yoshimura K, OtawaraHamamoto Y, Harrii K. Heterotopic ossification ofdegenerating rat skeletal muscle induced by adenovirus-mediated transfer of bone morphogenetic protein-2 gene. J Bone Miner Res 15, 1056-1065, 2000.
18. Yoshimura K, Harii K, Aoyama T, Iga T. Experience
with a strong bleaching treatment for skin hyperpigmentation in Orientals. Plast Reconstr Surg 105,
1097-1108, 2000.
19. Yoshimura K, Ouchi K, Wakita S, Uda K, Harii K.
Surgical correction of cryptotia with superiorly based
superficial mastoid fascia and skin paddle. Plast
Reconstr Surg 105, 836-841, 2000.
20. Kimata Y, Uchiyama K, Ebihara S, Kishimoto S, Asai
M, Saikawa M, Ohyama W, Haneda T, Hayashi R,
Onitsuka T, Nakatsuka T, Harii K. Comparison of
innervated and noninnervated free flaps in oral
reconstruction. Plast Reconstr Surg104, 1307-1313,
1999.
21. Shimoda Y, Tajima Y, Nagase T, Harii K, Osumi N,
Sanai Y. Cloning and expression of a novel galactoside beta1, 3-glucuronyltransferase involved in thebiosynthesis of HNK-1 epitope. J Biol Chem 274,
17115-17122, 1999.

22. Kimata Y, Uchiyama K, Sekido M, Sakuraba M, Iida
H, Nakatsuka T, Harii K. Anterolateral thigh flap for
abdominal wall reconstruction. Plast Reconstr Surg
103, 1191-1197, 1999.
23. Nakatsuka T, Harii K, Takushima A, Yoshimura K,
Ichioka S, Sugasawa M, Ichimura K, Seto Y, Nagawa
H. Prefabricated free jejunal transfer: a new reconstructive technique for high pharyngeal defects. Plast
Reconstr Surg103, 458-64, 1999.
24. Harii K. Asato H. Nakatsuka T. Ebihara S.
Reconstructive plastic surgery in cancer treatment:
surgery for quality of life. Jap Soc Clin Oncol 4, 193201, 1999.
25. Yoshimura K, Harii K. A regenerative change during
muscle adaptation to denervation in rats. J Surg Res
81, 139-146, 1999.
26. Gonda K, Okamoto H, Takuwa N, Yatomi Y, Okazaki
H, Sakurai T, Kimura S, Sillard R, Harii K, Takuwa Y.
The novel sphingosine 1-phosphate receptor AGR16
is coupled via pertussis toxin-sensitive and -insensitive G-proteins to multiple signalling pathways.
Biochem J 337, 67-75, 1999.
27. Harii,K. : Free groin flap reconstruction of severeburn neck constructure. Grabb’s encyclopedia offlaps second edition (eds. Strauch,B. et al.) vol.1,
pp.478-480, Lippincott-Raven, Philadelphia, 1998.
28. Harii,K. Asato,H. : Microneurovascular free gracilis
muscle transfer for facial reanimation. Grabb’s encyclopedia of flaps second edition (eds. Strauch,B. etal.) vol.1, pp.521-524, Lippincott-Raven, Philadelphia,
1998.
29. Harii,K. : Microvascular free transfer of the groin skinflap. Grabb’s encyclopedia of flaps second edition(eds. Strauch,B. et al.) vol.II, pp.1110-1111,
Lippincott-Raven, Philadelphia, 1998.
30. Harii,K. : Microvascular free transfer of a groin skinflap. Grabb’s encyclopedia of flaps second edition(eds. Strauch,B. et al.) vol.III, pp.1328-1330,
Lippincott-Raven, Philadelphia, 1998.
31. Hirabayashi S, Sugawara Y, Sakurai A, Harii K, Park
S. Frontoorbital advancement by gradual distraction.
Technical note. J Neurosurg 89, 1058-61, 1998.
32. Kimata Y, Uchiyama K, Ebihara S, Nakatsuka T, Harii
K. Anatomic variations and technical problems of theanterolateral thigh flap: a report of 74 cases. Plast
Reconstr Surg 102, 1517-1523, 1998.
33. Yoshimura K, Harii K, Asato H, Ueda K, Yamada A.
Immunohistochemical analysis of clinically transplanted muscles. J Surg Res 79, 31-38, 1998.
34. Harii K, Asato H, Yoshimura K, Sugawara Y,
Nakatsuka T, Ueda K. One-stage transfer of the latissimus dorsi muscle for reanimation of a paralyzed
face: a new alternative. Plast Reconstr Surg 102, 941951, 1998.
35. Takushima A, Kitano Y, Harii K. Osteogenic potentialof cultured periosteal cells in a distracted bone gap in
rabbits. J Surg Res 78, 68-77, 1998.
36. Yoshimura K, Kuzon WM, Harii K. Myosin heavy
chain expression in skeletal muscle autografts underneural or aneural conditions. J Surg Res 75, 135-147,
1998.
37. Ueda K, Harii K, Asato H, Yamada A. Neurovascular
free muscle transfer combined with cross-face nerve
grafting for the treatment of facial paralysis in children. Plast Reconstr Surg 101, 1765-1773, 1998.
38. Ichioka S, Nakatsuka T, Sato Y, Shibata M, Kamiya
A, Harii K. Amrinone, a selective phosphodiesterase
III inhibitor, improves microcirculation and flap survival: a comparative study with prostaglandin E1. J
Surg Res 75, 42-48, 1998.
39. Kitano Y, Kurihara H, Kurihara Y, Maemura K, Ryo Y,
Yazaki Y, Harii K. Gene expression of bone matrix
proteins and endothelin receptors in endothelin-1deficient mice revealed by in situ hybridization. J
Bone Miner Res 13, 237-244, 1998.
40. Nakaoka T, Gonda K, Ogita T, Otawara-Hamamoto
Y, Okabe F, Kira Y, Harii K, Miyazono K, Takuwa Y,
Fujita T. Inhibition of rat vascular smooth muscle
proliferation in vitro and in vivo by bone morphogenetic protein-2. J Clin Invest 100, 2824-2832, 1998.
41. Ichioka S, Shibata M, Kosaki K, Sato Y, Harii K,
Kamiya A. Effects of shear stress on wound-healing
angiogenesis in the rabbit ear chamber. J Surg Res
72, 29-35, 1997.
42. Ueda K, Harii K, Yamada A. Free vascularized double
muscle transplantation for the treatment of facial
paralysis. Plast Reconstr Surg 95, 1288-1296, 1995.
43. Yamada A, Harii K, Ueda K, Asato H, Tanaka H.
Versatility of a cross-leg free rectus abdominis flap
for leg reconstruction under difficult and unfavorable
conditions. Plast Reconstr Surg 95, 1253-1257, 1995.
44. Kimata Y, Uetake M, Tsukada S, Harii K. Follow-up
study of patients treated for ingrown nails with thenail matrix phenolization method. Plast Reconstr
Surg 95, 719-724, 1995.
45. Ueda K, Harii K, Yamada A. Electromyographic studyof functional recovery of free vascularized muscles
grafted to the face. Plast Reconstr Surg 95, 296-304,
1995.
46. Kawashima T, Yamada A, Ueda K, Asato H, Harii K.
Tissue expansion in facial reconstruction. Plast
Reconstr Surg 94, 944-950, 1994.
47. Ueda K, Harii K, Yamada A. Free neurovascular muscle transplantation for the treatment of facial paralysis using the hypoglossal nerve as a recipient motor
source. Plast Reconstr Surg 94, 808-817, 1994.
48. Komuro Y, Takato T, Harii K, Yonehara Y. The histologic analysis of distraction osteogenesis of themandible in rabbits. Plast Reconstr Surg 94, 152-159,
1994.
49. Ueda K, Harii K, Yamada A. Long-term follow-up of
nerve conduction velocity in cross-face nerve grafting
for the treatment of facial paralysis. Plast Reconstr
Surg 93, 1146-1149, 1994.
50. Nakatsuka T, Harii K, Yamada A, Asato H, Ebihara S.
Versatility of a free inferior rectus abdominis flap forhead and neck reconstruction: analysis of 200 cases.
Plast Reconstr Surg 93, 762-769, 1994.

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Department of Oral andMaxillofacial Surgery
Outline and Research Objectives
Department of Oral and Maxillofacial Surgery, commenced by Dr. Hisashi Ishihara in
1900, is one of the oldest departments in Graduate School of Medicine, University of Tokyo.
Professor Takato is the 7th professor in our department. The department has been the centerof the clinical and basic research in oral and maxillofacial region in Japan. Congenital anomalies, jaw deformities, benign and malignant oral tumors, facial bone fractures, temporomandibular joint diseases, and dental treatment in systemic diseases are mainly treated andresearched in our department. Multidisciplinary treatment teamed by plastic surgeons, oral
surgeons, orthodontists, prothodontists, etc. is characteristic and has performed excellent
results in clinical works. In research fields, we have mainly performed the experimental
studies on the regenerative capacity of tissues, especially bone, periosteum, cartilage, perichondrium, vessels, nerve, skin, etc. At present, we are focusing on tissue engineering inresearch works especially in bone, cartilage, vessels. Professor Takato has established
Division of Tissue Engineering in Tokyo University Hospital and our department has two
endowment departments: Department of Cartilage and Bone Regeneration (Menicon
Co.,Ltd.) and Department of Clinical Vascular Regeneration (Daiichi Pharmaceutical Co.,
Ltd.) in Tissue Engineering Division. These departments have 1 associate professor, 1staff,
and 4 graduate students in each. These staff are focusing on translational research works inmaxillofacial regions.

Research Objectives
1) Multidisciplinary treatment of facial deformities in patients with cleft lip and palate orother congenital maxillofacial anomalies
2) Multidisciplinary treatment of dentomaxillofacial deformities, fractures and temporomandibular diseases3) Multidisciplinary treatment of malignant tumors in head and neck region4) Reconstruction using bone and cartilage grafts in maxillofacial region5) Clinical and research works on distraction osteogenesis in maxillofacial region6) Basic research of bone metabolism7) Basic research on osteogenic capacity of periosteum8) Basic research on capacity of perichondrial regeneration
9) Basic research on growth plate
10) Tissue engineering on bone, cartilage, and vessels, etc

Faculties and Students
Professor and Chair

Tsuyoshi Takato, M.D., PhD.
(since 1996)
Associate Professor Takafumi Susami, D.D.S., PhD.
Lecturer
Yoshiyuki Mori, D.D.S., PhD.
Tomoaki Eguchi, M.D., PhD.
Tetsuya Yoda, D.D.S., PhD.
Hisako Hikiji, D.D.S., PhD.
Associate ...............................10
Clinical Stuff............................7
Resident ..................................9
Graduate student ..................12
Research student.....................1

258

Secretary .................................2

Past Research and Major Accomplishments
1. Clinical research:
1) Multidisciplinary treatment of facial deformities
in patients with cleft lip and palate or other congenital maxillofacial anomalies
2) Multidisciplinary treatment of dentomaxillofacial
deformities, fractures and temporomandibulardiseases
3) Multidisciplinary treatment of malignant tumors
in head and neck region4) Facial bone lengthening by distraction osteogenesis5) Correction of facial deformity in cleft lip andpalate patients
6) Speech problems in cleft lip and palate patients
7) Facial growth in craniofacial anomalies
8) Evaluation of the treatment outcomes in patientswith cleft lip and/or palate
9) Combined surgical-chemical-radiological treatment
for malignant tumors
10) Development of dental implant
11) Surgical-orthodontic treatment of dentofacial
deformities
12) Effects of arthrocentesis or therapeutic exercisesfor temporomandibular disorders
13) Effects of masticatory force on facial form
14) Non-surgical treatment system for facial bone fractures
2. Basic and experimental research:
1) Osteogenic capacity of periosteum2) Capacity of perichondrial regeneration
3) Osteogenic capacity of growth plate4) Development of various types of new skin flaps
5) Metabolism of poly ADP-ribose in DNA repair andcell differentiation
6) Gene analysis of congenital anomalies of oral andmaxillofacial region7) Effect of free radicals on bone metabolism
8) Intracellular calcium handling on osteoblasts9) Differentiation mechanism of osteoblasts in termsof cell cycle molecule10) Functional analysis on domains of P130Cas in actincytoskeleton organization, cell migration, and
Src transformation
11) Osteochondrogenic differentiation of bone marrow derived mesenchymal stem cells by spheroid
culture12) Cloning of the 5’Upstream Region of the Rat p16
Gene and Its Role in Silencing
13) Studies on relationship between dystrophin associated protein and advanced heart failure14) Mandibular lengthening by floating bone method
15) Periodontal tissue regeneration on dental implants16) Characterization of skin derived multipotent stem
cells, especially differentiation mechanism into
neuronal cell17) Bone and cartilage repair in dentomaxillofacial
region using tissue engineering technique259

Current ResearchMultidisciplinary treatment
Transplantation (carterige,bone,etc)

Oral surgeon

Distraction osteogenesis

Plastic surgeon

Dental Implant

Orthodontists

Orthognathic Surgery
Orthodontic treatment

Prothodontist

Cancer treatment

Translational Research
Tissue Engineering
Division of Tissue Engineering
Bone and Cartilage Regenerative Medicine (Takeda Chemical Industories, Ltd.)
Cartilage and Bone Regeneration (Menicon Co.,Ltd.)

School of Enginnering
Scaffold

Clinical Vascular Regeneration (Daiichi Pharmaceutical Co., Ltd.)Drug delivery system

Hematopoietic Regeneration(Kirin Brewery Company, Limited)

School of medicine
Clinical Renal Regeneration(Mochida Pharmaceutical Co.,Ltd.)

Corneal Tissue Regeneration(HOYA Healthcare Corporation)

Internal Medicine
Vascular Surgery
Orthopedic Surgery
Gene Therapy
Drug delivery system
etc.

International Medical Center Research InstituteNational Cancer Center(Gene therapy, Stem Cell)

Kitazato Medical School (Stem Cell)
National Institute of Advanced Industrial Science and Technology
KOKEN Co.,Ltd260

Future Prospects
Our department has been the center of the treatment and research in dentomaxillofacial diseases forthe last century in Japan. For decade years, we established the system of the multidisciplinary treatment
for diseases in dentaomaxillofacial region. We are
now in the next step to develop the new treatment
for deficiency of dentomaxillofacial region using tissue-engineering technique. For this purpose, Professor
Takato established the Division of Tissue Engineeringin Tokyo University Hospital in 2001. We have been
researching the development of the materials, tissues
and organs which are constructed with the tissueengineering technique. Three-dimensional construction of cartilage and bone with neovascularization is
our most important theme to investigate. These
regenerated tissues would repair the deficiency of thedentomaxillofacial region without the harvest of grafted materials from the donor sites. To achieve thistranslational research, we are cooperating with several
companies and Ministries.

Research Grants
The Ministry of Edition, Science, Culture and Technology
Research Grant B2 (1995-1996)
Research Grant B1 (1997-1998)
Research Grant B2 (1997-1998)
Research Grant B2 (1999−2001)
Grant for development of advanced medicine (2002-2004)

Select Publications
1. Harii K., Ebihara S., Ono I., Saito H., Terui S., andTakato T.: Pharyngoesophageal reconstruction usinga fabricated forearm free flap. Annals of Plastic
Surgery, 85: 463-474, 1985
2. Takato T., Ono I., Ebihara S., and Saeki T.: Repair of
stenosed tracheostoma. Journal of Oto-RhinoLaryngology & its Related Specialties, 47: 299-302,
1985
3. Takato T., Ono I., Ebihara S., and Matsumoto T.:
Reconstruction of the oral commissure. Oral Surgery,
Oral Medicine, Oral Pathology, 62: 132-134, 1986
4. Takato T., Harii K., Nakatsuka T., Ueda K., andOhtake T.: Vascularized periosteal graft : An experimental study using two different forms of tibial
periosteum in rabbits. Plastic and Reconstructive
Surgery, 78: 489-497, 1986
5. Takato T., Harii K., Ebihara S., Ono I., Yoshizumi T.,
and Nakatsuka T.: Oral and pharyngeal reconstruction using the free forearm flap. Archives of
Otolaryngology, 113: 873-879, 1987
6. Takato T., Harii K., and Nakatsuka T.: The development of bone after perichondrial grafting : An experimental study using ear and rib perichondrium in rab-

bits. British Journal of Plastic Surgery, 40: 636-641,
1987
7. Takato T., Harii K., and Nakatsuka T.: The sequential evaluation of bone scintigraphy : an analysis ofrevascularised bone grafts. British Journal of Plastic
Surgery, 41: 262-269, 1988
8. Takato T., Harii K., and Nakatsuka T.: Osteogenic
capacity of vascularized periosteum : experimental
study using rib periosteum in rabbits. British Journalof Plastic Surgery, 41: 528-532, 1988
9. Takato T., Harii K., Nakatsuka T., and Ohtake T.:
Experimental study of vascularized bone :Quantitative analysis of bone scintigraphy and histology. Journal of Reconstructive Microsurgery, 4:391-397, 1988
10. Takato T., Ohsone H., and Tsukakoshi H.:
Treatment of severe microstomia by swallowing
caustic soda. Oral surgery, Oral medicine, Oralpathology, 67: 20-24, 1989
11. Takato T., Kamei M., Kato K., and Kitano I.: Cleft
palate in the Beckwith-Wiedemann syndrome.
Annals of Plastic Surgery, 22: 347-349, 1989
12. Takato T., Itoh M., Yokota M., Kyoku I., Kitano M.,
Misuhara T., and Sakamoto K.: Delayed thoracic closure after cardiac surgery in infants. British Journalof Plastic Surgery, 44: 106-108, 1991
13. Takato T., Zuker R.M., and Turley C.B.:
Prefabrication of skin flaps using vein graft : anexperimental study in rabbits. British Journal of
Plastic Surgery, 44: 593-598, 1991
14. Takato T., Nakai H., Yonehara Y., Takeda H., Susam
P., Kitano Y., and Kato K.: Binder’s syndrome :Peculiarities in Japanese patients. Annals of Plastic
Surgery, 27: 371-381, 1991
15. Takato T., Zuker R.M., and Turley C.B: Viability andversatility of arterialized venous perfusion flaps andprefabricated flaps : an experimental study in rabbits. Journal of Reconstructive Microsurgery, 8: 111119, 1992
16. Takato T., Komuro Y., Yonehara Y., and Zuker R.M.:
Viability and versatility of arterialized venous perfusion flaps and prefabricated flaps : an experimental
study in rabbits. British Journal of Plastic Surgery,48: 111-119, 1993
17. Takato T., Komuro Y., Yonehara Y., and Zuker R.M.:
Prefabricated venous flaps : an experimental study in
rabbits. British Journal of Plastic Surgery, 46: 122126, 1993
18. Takato T., Harii K., Komuro Y., and Yonehara Y.:
Experimental study on growth of epiphysial plate :free graft in rabbits. British Journal of Plastic Surgery,46: 416-420, 1993
19. Komuro Y., Takato T., Yamada A., and Yonehara Y.:
Experimental study of prefabricated flaps using vein
grafts. Journal of Reconstructive Microsurgery, 9:373-380, 1993
20. Takato T., Harii K., Komuro Y., Yonehara Y., andSusami T.: Mandibular lengthening by gradual distraction : analysis using accurate skull replicas.
British Journal of Plastic Surgery, 46: 686-693, 1993

261

21. Komuro Y., Takato T., Harii K., and Yonehara Y.:
The histological analysis of distraction osteogenesisof mandible in rabbits. Plastic and Reconstructive
Surgery, 94: 152-159, 1994
22. Takato T., Harii K., Yonehara Y., Komuro Y., Susami
T., and Uoshima K.: Correction of the cleft nasal
deformity with an L-shaped iliac bone graft. Annalsof Plastic Surgery, 33: 486-493, 1994
23. Komori Y., Takato T., and Akagawa T.: Use of alaser-hardened three dimensional replica for simulated surgery. Journal of Oral and Maxillofacial Surgery,52: 516-521, 1994
24. Takato T., Yonehara Y., and Susami T.: Early correction of the nose in unilateral cleft lip patients using
an open method : A 10-year review. Journal of Oraland Maxillofacial Surgery, 53: 28-33, 1995
25. Takato T., Yonehara Y., and Susami T.: Columella
lengthening using a cartilage graft in the bilateralcleft lip-associated nose : Choice of cartilage according to age. Journal of Oral and Maxillofacial Surgery,53: 149-157, 1995
26. Takato T., Yonehara Y., Mori Y., Sakamoto Y., andSusami T.: Use of Cantilever Iliac Bone Grafts for
Reconstruction of Cleft Lip-Associated Nasal
Deformities. Journal of Oral and Maxillofacial
Surgery, 53: 757-762, 1995
27. Komori T., Takato T., and Yokoyama K.: Integrationof dental models in a three-dimensional replica.
Journal of Oral and Maxillofacial Surgery, 53: 621624, 1995
28. Komori T., Matsumoto K., Yokoyama K., and Takato
T.: A Morphological Study of Bone Cuts with
Aluminium Oxide Particles. Japanese Journal of Oral
Diagnosis/ oral Medicine. 8: 208-212, 1995
29. Takato T., Yonehara Y., Susami T., and Uoshima K.:
Modification of the Abbe Flap for Reconstruction fo
the Vermilion Tubercle and Cupid’s Bow in Cleft Lip
Patients. Journal of Oral and Maxillofacial Surgery,
54: 256-261, 1996
30. Komori T., Yokoyama K., Takato T., and Matsumoto
K.: Clinical application of the erbium:YAG laser forapicoectomy. Journal of Endodontics, 23:748-750，
1997
31. Nozaki T., Masutani M., Sugimura T., Takato T., andWakabayashi K.: Abrogation of G1 arrest after DNA
damages is associated with constitutive expressionof Mdm2, Cdk4, and Irf1 mRNAs in the BALB/c 3T3
A31 Variant 1-1 Clone. Biochemical & BiophysicalResearch Communications, 233: 1997
32. Hikiji H., Wee Soo Shin, Oida S., Takato T, Koizumi
T, and Toyooka T: Direct action of nitric oxide on
osteoblastic differentiation. FEBS Letters, 410:238242, 1997
33. Yonehara Y., Takato T., Harii K., Hirabayashi S.,
Susami T., Komori T., Matsumoto S., Hikiji H., andNakatsuka T.: Secondary lengthening of the reconstructed mandible using a gradual distraction technique-two case reports. British Journal of Plastic
Surgery, 51: 356-358, 1998

262

34. Nakatsuka T., Harii K., Yamada A., Yonehara Y.,
Takato T., Kawahara N., Sakaki T., Yamasoba T.,
Nibu K., and Ebihara S.: Immediate Free Flap
Reconstruction for Head and Neck Pediatric
Malignancies. Annals of Plastic Surgery, 40(6): 594599, 1998
35. Sakata Y., Tokunaga K., Yonehara Y., Bannai M.,
Tsuchiya N., Susami T., and Takato T.: Significant
association of HLA-B and HLA-DRB1 alleles withcleft lip with or without cleft palate. Tissue
Antigens, 53: 147-152, 1999
36. Takushima A., Susami T., Nakatsuka T., Harii K.,
and Takato T.: Multi-Bracket Appliance inManagement of Mandibular Reconstr uction with
Vascularized Bone Graft. Japanese Journal of ClinicalOncology, 29: 119-126, 1999
37. Yonehara Y., Takato T., Matsumoto S., Nakatsuka
T.: Distraction of scarred soft tissue before secondarybone grafting. International Journal of Oral
Maxillofacial Surgery, 28: 347-348, 1999
38. Yonehara Y., Nakatsuka T., Ichioka I., Takato T.,
Matsumoto S., and Yamada A.: Intramuscular haemangioma of the anterior chest wall. British Journalof Plastic Surgery, 53: 257-259，2000
39. Yonehara Y., Takato T., Matsumoto S., Mori Y.,
Nakatsuka T., and Hikiji H.: Correction of cleft lip
nasal deformity in Orientals with a cantilevered iliacbone graft. Scandinavian Journal of Plastic andReconstructive Surgery and Hand Surgery, 34: 137143，2000
40. Yonehara Y., Takato T., Susami T., and Mori Y.:
Correction of Micrognathia Attributable to Ankylosisof the Temporomandibular Joint Using a GradualDistraction Technique: Case Report. Journal of Oraland Maxillofacial Surgery, 58:1415-1418，2000
41. Chikazu D., Shindo M., Iwasaka T., Katagiri M.,
Manabe N., Takato T., Nakamura K., and Kawaguchi
H.: A Novel Synthetic Triazolotriazepine Derivative
JTT-606 Inhibits Bone Resorption by DownRegulation of Action and Production of Bone
Resorptive Factors. Journal of Bone Miner Research,
15(4):674-682，2000
42. Hikiji H., Shin WS, Koizumi T., Takato T., Susami T.,
Koizumi Y., Okai-Matsuo Y., and Toyooka T.:
Peroxynitrite production by TNF-alpha and IL-1beta:
implication for suppression of osteoblastic differentiation. American Journal of Physiology Endocrinology & Metabolism, 278: E1031-1037，
2000
43. Hikiji H., Takato T., Matsumoto S., and Mori Y.:
Experimental Study of Reconstruction of the
Temporomandibular Joint Using a Bone Transport
Technique. Journal of Oral and Maxillofacial Surgery,
58: 1270-1276，2000
44. Chikazu D., Hakeda Y., Nemoto K., Itabashi A.,
Takato T., Kumegawa M., Nakamura K., andKawaguchi H.: Fibroblast growth factor-2 directly
stimulates mature osteoclasts activity through autophosphorylation of FGF receptor 1. Journal of
Biological Chemistry2000, 275: 31444-31450，2000

45. Katagiri M., Hakeda Y., Chikazu D., Ogasawara T.,
Takato T., Kumegawa M., Nakamura K., andKawaguchi H. :Mechanism of Stimulation of
Osteoclastic Bone Resorption through Gas6／Tyro3,
a Receptor Tyrosine Kinase Signaling in Mouse
Osteoclasts. Journal of Biological Chemistry2001,
276(10):b7376-7382, 2001
46. Eguchi T., Asato H., Takushima A., Takato T., andHarii K. : Surgical repair for congenital macrostomia:
A vermilion square flap method. Annals of Plastic
Surgery 47:629-635, 2001
47. Chikazu D., Katagiri M., Ogasawara T., Ogata N.,
Shimoaka T., Takato T., Nakamura K., andKawaguchi H.: Regulation of osteoclast differentiation by fibroblast growth factor 2: Stimulation ofreceptor activator of nuclear factorκB ligand/osteoclast differentiation factor expression in osteoblasts
and inhibition of macrophage colony-stimulating factor function in osteoclast precursors. Journal of Bone& Mineral Research, 16(11): 2074-2081, 2001
48. Sakai N., Tokunaga K., Yamazaki Y., Shida H., Sakata
Y., Susami T., Takato T. and Uchinuma E. : Sequence
analysis of fibroblast growth factor receptor 2
FGFR2) in Japanese patients with craniosynostosis.
Journal of Craniofacial Surgery, 12: 580-585, 2001
49. Nozaki T., Fujihara H., Kamada N., Ueda O., TakatoT., Nakagama H., Sugimura T., M.J.A., Suzuki H.,
and Masutani M. ：Hyperploidy of embryonic
fibroblasts derived from Parp-1 knockout mouse.
Proceeding of the Japan Academy, 77B(6) : 121124,2001
50. Jinhong Huang, Hamasaki H., Nakamoto T., Honda
H., Hirai H., Saito M., Takato T., Sakai R. :Differential regulation of cell migration, actin stressfiber organization and cell transformation by functional domains of Cas Journal of Biological Chemistry277(30): 27265-27272, 2002

263

Department ofOrthopaedic Surgery
Outline and Research Objectives
Our department was established in 1906 as the first department of orthopaedic surgery
in Japan. Since then our efforts have been dedicated to responding to the needs of patientsfor orthopaedic care and to related research. We have pioneered handicapped children’scare, developed new and innovative surgical techniques, and invented arthroscopy which is
now used worldwide.
Our department seeks to elucidate the molecular and genetic backgrounds of bone andcartilage disorders including osteoporosis, osteoarthritis, and joint destruction in rheumatoid arthritis. To achieve this, we utilize the newest methodologies, and with the knowledge gained, we are working to develop groundbreaking treatments for these conditions,such as bone and cartilage regeneration. At the same time we are attempting to establish a
non-invasive analyzing system to evaluate the mechanical property of the skeletal system
and a computer-guided operating system to put to use our findings of the last half decade.

Faculty and Students
Professor and Chair

Kozo Nakamura, M.D., Ph.D.
(1998-)
Associate Professor Hiromi Oda, M.D., Ph.D.
Yoshio Takatori, M.D., Ph.D.
Lecturer
Isao Ohnishi, M.D., Ph.D.
Atsushi Seichi, M.D., Ph.D.
Takahiro Goto, M.D., Ph.D.
Hiroshi Kawaguchi, M.D., Ph.D.
Hiroshi Okazaki, M.D., Ph.D.
Associate ...............................12
Postdoctoral Fellow ................1
Graduate student ..................25
Secretary .................................3

Past Research and Major Accomplishments
1. Regulation of anabolic and catabolic bonemetabolism by fibroblast growth factor-2 (FGF-2)
We have had achieved notable advances in laboratory and clinical research on cytokines / growth factors in bone. Perhaps our most important contribution has been in defining the actions of FGF-2 onbone. We have reported the anabolic action of FGF-2
on bone formation using several physiological andpathological animal models of both rodents and primates. Our efforts in this area have led to studies ofclinical application of FGF-2 to fracture healing andosteoporosis. The involvement of FGF-2 on bone
resorption has also been investigated and this may
possibly be involved in the joint destruction byrheumatoid arthritis.

264

2. Molecular mechanism of age-related bone loss
(A novel aging suppressor gene: klotho)
Reverse genetic analysis revealed that mice deficient in the klotho gene exhibited bone loss with low
turnover which is characteristic of human osteoporosis with aging. Using a forward genetic approach, we
found that two polymorphisms at the human klotho
locus are correlated with bone density in postmenopausal women, especially in their older years
and independent of race, suggesting that the klotho
gene may be involved in bone loss with aging.
3. Involvement of insulin signaling in bone anabolic action
Through clinical studies we found that the severityof ossification of the posterior longitudinal ligamentof the spine (OPLL) is associated with insulin secretory response due to impairment of the response toinsulin in its target tissues. The importance of insulin
signaling in the anabolic bone metabolism was confirmed by reverse genetic approach on knock-out miceof the insulin receptor substrates IRS-1 and IRS-2,
which are essential intracellular signaling molecules of
insulin. It was shown that IRS-1 maintains boneturnover while IRS-2 is needed to maintain the predominance of bone formation over bone resorption;
the integration of these two signalings causes a potent
bone anabolic action by insulin.
4. Molecular mechanism of osteoclast differentiation in rheumatoid arthritis (RA)Recent studies have revealed that a TNF-α superfamily cytokine, receptor activator of nuclear factor-kB
ligand (RANKL) plays a pivotal role in osteoclast differentiation and activation. We found that RANKL ishighly expressed in synovial fibroblasts obtained
from RA patient synovial tissues, and critically implicated in the bone and joint destruction of RA.
5. Gene transfer to osteoclasts and gene therapy
We recently established a gene transduction system into mature osteoclasts using adenovirus vectors,
and successfully regulated osteoclast function byintroducing several genes that modulate the function
and survival. Protooncogene product c-Src is a nonreceptor type tyrosine kinase which is indispensablefor osteoclast activation. The kinase activity of c-Src isregulated by another cytoplasmic tyrosine kinase Csk(C-terminal Src kinase). Introduction of csk gene into
osteoclasts by the adenovirus vector completely disrupted the cytoskeletal organization of the cells, andstrongly suppressed their bone-resorbing activity.
Using the adenovirus vector-mediated gene transduction system, we also found that the Ras-ERK pathway
is critical for the survival of osteoclasts.
6. Gene therapy of arthritic joint destruction
using adenovirus vectors
Adenovirus vectors are effective not only in vitro
but also in vivo gene delivery into osteoclasts. Local
injection of Csk adenovirus reduced IL-1-inducedbone destruction in calvarial bone. When injected into
arthritic joints, adenovirus vectors efficiently transduce synovial fibroblasts as well as osteoclasts. Csk
adenovirus inhibited proliferation of synovial fibroblasts and their proinflammatory cytokine production
in vitro, and intraarticular injection of the virus significantly ameliorated joint destruction in adjuvant
arthritis rats. These results demonstrate that the adenovirus vector system is a good therapeutic approachto arthritic bone destruction.
We also found that adenovirus vector-mediatedcsk gene transduction suppressed the tumor metastasis and constitutively active MEK1 gene recovered theparaplegia after spinal cord injury.
7. Regulation of bone homeostasis by RANKL and
IFNs (interferons)
Investigation of the regulation of osteoclastogenesis by the immune system showed the critical involvement of IFNs in bone metabolism. Mice deficient in
IFNGR1 (IFN-γ receptor) exhibited more severe bonedamage accompanied by enhanced osteoclastogenesis
when stimulated with lipopolysaccharide (LPS).
Activated T cells strongly inhibited osteoclastogenesis
via IFN-γ, and IFN-γ interfered with RANKL signaling
by inhibiting expression of TRAF6 (TNF receptor
associated factor 6). Thus, IFN-γ protects against T
cell-mediated inflammatory bone loss. In contrast,
mice deficient in IFNAR1 (IFN-α receptor, one of IFNα/β receptor components) spontaneously develop

marked osteopenia accompanied by enhanced osteoclastogenesis. RANKL induces IFN-β gene in osteoclast precursor cells and IFN-β inhibits the differentiation by interfering with the RANKL-induced expression of c-Fos, an essential transcription factor for
osteoclastogenesis. Thus, IFN-β acts as a negative
feedback regulator of RANKL-induced osteoclastogenesis in physiological bone remodeling. Thus, IFN-γ
and IFN-β, both of which play pivotal roles in the
immune system, also function in the negative regulation of RANKL signaling via distinct mechanisms.

Current Research
1. Reverse and forward genetic approach to thepathophysiology of osteoarthritis
Osteoarthritis, a major skeletal disorder, is considered a collective result of heterogeneous etiopathologic factors affecting cartilage. We actually identified
two novel genes associated with chondrocyte differentiation and degradation: cystacin10 and cgk2. To
further determine the molecular background of thedisorder, we created an osteoarthritis model in miceby cutting ligaments in and around the knee joint.
Several knock-out or transgenic mice were subjected
to an operation to learn the involvement of themanipulated gene in the disorder. In addition, as aforward genetic approach, we are in the middle of the
genome-wide association study on the human knee
and hip osteoarthritis.
2. Gemone-wide association study of OPLL
Based on a reverse genetic study, we have foundthat nucleotide pyrophosphate (Npps) gene is related
to the incidence and progression of OPLL. Since OPLL
is a polygenic disease, we are conducting a genomewide association study of the condition in whichmore than 100,000 single nucleotide polymorphisms(SNPs) will be evaluated for their association with susceptibility to and severity of OPLL. DNA samples are
now being collected in a national project headed by
the presenter.
3. Molecular mechanism of age-related bone loss
(PPAR-γ)Considering that osteoblasts and adipocytes shareorigins in bone marrow mesenchymal cells, and thathuman aging is associated with a reciprocal increasein adipocytes and a decrease in osteoblasts in bonemarrow, PPAR-γ may be clinically involved in thepathophysiology of bone loss with aging as a suppressor of osteoblastogenesis. To identify the role ofendogenous PPAR-γ in vivo, we examined the bone of
PPAR-γ hetero knock-out mice, and found that bonemass in these mice increases with stimulation of265osteoblastogenesis from bone marrow progenitors
without affecting mature osteoblasts or osteoclast-lineage cells. The forward genetic approach beginningwith the screening of polymorphisms of the human
PPAR-γ gene is now underway.
4. Cloning master gene(s) of osteoclast differentiation
In an attempt to identify gene(s) that regulate
osteoclast differentiation, we are utilizing a molecularindexing technique, and have found several genesthat regulate osteoclast-specific promoter activity. We
are currently working on the function of the genes inosteoclasts.
5. Treating arthritic bone destruction by targeting
RANKL/RANK pathways
As mentioned above, RANKL/RANK pathways are
critically involved in pathological bone resorption,and therefore can be a good therapeutic target. The
effect of osteoprotegerin (OPG), a natural inhibitor of
RANKL, on pathological bone resorption is currently
being examined. In a preliminary study, local injection of recombinant human OPG into arthritic joints
effectively ameliorated the bone destruction in adjuvant arthritis rats.
We are also trying to establish a technology of
RANKL vaccination therapy, in which modified
RANKL is utilized to induce anti-RANKL self-antibody, which has neutralizing activity against self
RANKL, and therefore has a therapeutic effect onbone destruction.
6. Molecular mechanism of osteoclast apoptosis
Osteoclasts are terminally differentiated non-proliferating cells with a very short life span. We are now
trying to identify molecule(s) that induce osteoclast
apoptosis, and have found that a pro-apoptotic Bcl-2
family member Bim plays an essential role. In ourstudies on the role of small GTP-binding proteins on
osteoclast survival, Rac1 transduction was found to
prolong the survival.
7. Gene therapy of arthritic bone resorption
We recently found that adenovirus vectors carrying dominant negative Ras gene (AxRasDN) strongly
suppressed the proliferation and proinflammatory
cytokine production of synovial fibroblasts. We are
currently investigating the effect of AxRas DN on
arthritic bone destruction, and in a preliminary study,
found that this virus could significantly suppress joint
destruction in adjuvant arthritis rats.
8. Evaluation of mechanical properties of skeletal
system
A system of analyzing mechanical properties of a266

skeletal system has been required to clinically evaluate the efficacy of osteoporosis treatment and to
decide on a method or term of fracture treatment. We
are developing a non-invasive analyzing system invivo using CT based finite element analyses which
can make it possible to predict fracture load and location, which is based on the maximum principal stressfailure theory.
9. Investigation of adult neural progenitors as a
new therapeutic tool for damaged spinal cord
Seeking a therapeutic tool useful in the treatmentof the damaged spinal cord, we have investigatedadult endogenous neural progenitors, including stem
cells. Their widespread occurrence and proliferationin response to injury have been demonstrated. One of
their regulatory mechanisms in vivo, Notch signaling,is suggested from their molecular property. Thus,
genetic manipulations of endogenous progenitors insitu may enable the recruitment of their latent regenerative potential. Further advances of such strategies
may make possible significant structural repair of thedamaged spinal cord.
10. Development of a joint prosthesis with lessloosening
We are developing a new type of hip joint prosthesis whose surface is covered with a synthetic material,methacryloyloxyethyl phoshorylcholine (MPC),
resembling the human membrane phospholipid. We
have confirmed that the prosthesis successfully
decreases the friction index up to 1/5 of the conventional one. Due to the similarity of the material to living membrane, this may possibly elicit a less rejective
response causing loosening. This prosthesis will
improve the outcome of joint replacement surgery
and lead to a new era in osteoarthritis treatment.
11. Navigation robot for minimally invasive surgery
Image-guided surgery is gradually spreading withthe advancement of pre-surgical simulation using Xray CT or MR images to achieve minimally invasive
and safer surgical procedures. In cervical bone fixation
surgery, highly accurate positioning accuracy isrequired during drilling to avoid injuring the spinal
cord and major blood vessels. However, when using
an image-guided navigation system, we encounter difficulties in obtaining accurate “registration” between
the navigation results and a device such as a needle,
screw or drill; these difficulties arise mainly either
from a surgeon’s tremor or complicated spatial orientation relations. To overcome these problems, wehave developed the first prototype of a simple robot
having two degrees-of-freedom (2-DOF), which controls the orientation of a device. The characteristics of

the robot, low DOF, restricted range of movement,
and light weight, make it possible to use it in a clinicalsetting.

Future Prospects
1. Bone and cartilage regeneration
We plan to introduce a new type of bone and cartilage regeneration based not only on tissue engineering, but also on molecular and genetic biology.
Endochondral bone development is characterized by aprocess in which a cartilage mold is replaced by bone.
During this process, cartilage induces bone in theadjacent tissue. We have found that a subgroup of cartilage containing hypertrophic chondrocytes plays anessential role in this induction. In addition, thesechondrocytes induce vascular invasion. Based onthese two findings, we came up with a new strategy
to regenerate bone, which is a relatively well vascularized organ. Instead of separately regenerating boneand vasculature and subsequently combining andtransplanting them to create viable bone tissue, wehave proposed to graft hypertrophic chondrocytes tohave them induce both bone and vasculature simultaneously. To realize this unique strategy, we are trying
to improve methods to derive chondrocytes from various stem cells, differentiate them further into hypertrophic chondrocytes, and combine them with appropriate scaffolds to create new osteogenic-angiogenic
biomaterial.

2. Control of pathological bone destruction
In the future, we will be able to establish novel
treatments for pathological bone destruction, and theRANKL/RANK pathway can be a good therapeutic target. RANKL vaccination technology is a promisingpreventive therapy. By identifying the genes and signal transduction pathways that regulate osteoclast differentiation, activation, and survival, we will be able
to regulate bone resorption much more precisely andmeticulously. Adenovirus vector-mediated gene transduction will be a good means of gene therapy targeting osteoclasts by modulating intracellular signals.
3. Surgical navigation system with intuitive threedimensional display and navigation robot
We are developing a new surgical navigation system which superimposes the real, intuitive threedimensional (3-D) image of the patient’s internal
structure on the patient’s body, and helps surgeons toperform surgery. The system consists of a personal
computer, a lens array, a supporting stage, a liquid
crystal display and a half-silvered mirror. The 3-D
images are generated by real-time computer-generated
integral photography, and superimposed on thepatient’s body via a half-silvered mirror, as if they
were seen through the body. Because of the simplicity
and the intuitiveness of the navigation image, thissystem will become applicable for clinical use in thenear future in combination with the navigation robot
system described above.

Research Grants (Figure 1)

267

Select Publications (50)
1. Nakamura K, Kurokawa T, Aoyama I, Hanada K,
Tamura M, Kawaguchi H: Stimulation of bone formation by intraosseous application of basic fibroblast
growth factor in ovariectomized rats. Int Orthop 22:
49-54, 1998
2. Tanaka S, Takahashi T, Takayanagi H, Miyazaki T,
Oda H, Nakamura K, Hirai H, Kurokawa T:
Modulation of osteoclast function by adenovirus vector-induced epidermal growth factor receptor. J Bone
Miner Res 13: 1714-1720, 1998
3. Nakamura K, Matsushita T, Mamada K, Okazaki H,
Ou W, Okuma Y, Kurokawa T: Changes of callus
diameter during axial loading and after fixator
removal in leg lengthening. Arch Orthop Trauma
Surg 117: 464-467, 1998
4. Kato T, Kawaguchi H, Hanada K, Aoyama I, Hiyama
Y, Kuzutani T, Tamura M, Kurokawa T, Nakamura K:
Single local injection of recombinant fibroblast
growth factor-2 stimulates healing of segmental bone
defects in rabbits. J Orthop Res 16: 654-659, 1998
5. Okazaki H, Kurokawa T, Nakamura K, Matsushita T,
Mamada K, Kawaguchi H: Stimulation of bone formation by recombinant fibroblast growth factor-2 incallotasis bone lengthening of rabbits. Calcif Tissue
Int 64: 542-546, 1999
6. Nakamura I, Ikegawa S, Okawa A, Okuda S,
Koshizuka Y, Kawaguchi H, Nakamura K, Koyama T,
Goto S, Toguchida J, Matsushita M, Ochi T, Takaoka
K, Nakamura Y. Association of the human NPPS
gene with ossification of the posterior longitudinal
ligament of the spine (OPLL). Hum Genet 104: 492497, 1999
7. Manabe N, Oda H, Nakamura K, Kuga Y, Uchida S,
Kawaguchi H: Involvement of fibroblast growth factor-2 in joint destruction of rheumatoid arthritis
patients. Rheumatology 38: 714-720, 1999
8. Takayanagi H, Juji T, Miyazaki T, Iizuka H,
Takahashi T, Isshiki M, Okada M, Tanaka Y,
Koshihara Y, Oda H, Kurokawa T, Nakamura K,
Tanaka S: Suppression of arthritic bone destruction
by adenovirus-mediated csk gene transfer to synoviocytes and osteoclasts. J Clin Invest 104: 137-146,
1999
9. Kawaguchi H, Manabe N, Miyaura C, Chikuda H,
Nakamura K, Kuro-o M: Independent impairment of
osteoblast and osteoclast differentiation in klotho
mouse exhibiting low-turnover osteopenia. J Clin
Invest 104: 229-237, 1999
10. Nishikawa T, Ogawa S, Kogita K, Manabe N,
Katsumata T, Nakamura K, Kawaguchi H: Additiveeffects of combined treatment with etidronate andalfacalcidol on bone mass and mechanical properties
in ovariectomized rats. Bone 27: 647-654, 2000
11. Takayanagi H, Iizuka H, Juji T, Nakagawa T,
Yamamoto A, Miyazaki T, Koshihara Y, Oda H,
Nakamura K, Tanaka S: Involvement of receptor activator of nuclear factor kB ligand/osteoclast differentiation factor in osteoclastogenesis from synoviocytes268

in rheumatoid arthritis. Arthritis Rheum 43: 259269, 2000
12. Kawaguchi H, Chikazu D, Nakamura K, Kumegawa
M, Hakeda Y: Direct and indirect actions of fibroblast growth factor-2 on osteoclastic bone resorption
in cultures. J Bone Miner Res 15: 466-473, 2000
13. Ogata N, Chikazu D, Kubota N, Terauchi Y, Tobe K,
Azuma Y, Ohta T, Kadowaki T, Nakamura K,
Kawaguchi H: Insulin receptor substrate-1 inosteoblast is indispensable for maintaining boneturnover. J Clin Invest 105: 935-943, 2000
14. Takayanagi H, Ogasawara K, Hida S, Chiba T,
Murata S, Sato K, Takaoka A, Yokochi T, Oda H,
Tanaka K, Nakamura K, Taniguchi T: T cell-mediatedregulation of osteoclastogenesis by signaling crosstalk between RANKL and IFN-γ. Nature 40: 600605, 2000
15. Nakagawa T, Tanaka S, Suzuki T, Takayanagi H,
Miyazaki T, Nakamura K. Tsuruo T: Overexpressionof the csk gene suppresses tumor metastasis in vivo.
Int J Cancer 88: 384-391, 2000
16. Miyazaki T, Takayanagi H, Isshiki M, Takahashi T,
Okada M, Fukui Y, Oda H, Nakamura K, Hirai H,
Kurokawa T, Tanaka S: In vitro and in vivo suppression of osteoclast function by adenovirus vectorinduced csk gene. J Bone Miner Res 15: 41-51, 2000
17. Miyazaki T, Katagiri H, Kanegae Y, Takayanagi H,
Sawada Y, Yamamoto A, Pando MP, Asano T, Verma
IM, Oda H, Nakamura K, Tanaka S: Reciprocal role of
ERK and NF-κB pathways in survival and activationof osteoclasts. J Cell Biol 148: 333-342, 2000
18. Miura T, Tanaka S, Seichi A, Arai M, Goto T, Katagiri
H, Asano T, Oda H, Nakamura K: Partial functional
recovery of paraplegic rat by adenovirus-mediatedgene delivery of constitutively active MEK1. Exp
Neurol 166: 115-126, 2000
19. Kawaguchi H, Manabe N, Chikuda H, Nakamura K,
Kuro-o K: Cellular and molecular mechanism of lowturnover osteopenia in the klotho-deficient mouse.
Cell Mol Life Sci 57: 731-737, 2000
20. Ohtsuka T, Ohnishi I, Nakamura K, Takamoto S:
New instrumentation for video-assisted anterior
spine release. Surg Endosc 14: 682-684, 2000
21. Manabe N, Kawaguchi H, Chikuda H, Miyaura C,
Inada M, Nagai R, Nabeshima Y, Nakamura K,
Sinclair AM, Scheuermann RH, Kuro-o M:
Connection between B lymphocyte and osteoclastdifferentiation pathways. J Immunol 167: 2625-31,
2001
22. Yamamoto N, Yamamoto S, Inagaki F, Kawaichi M,
Fukamizu A, Kishi N, Matsuno K, Nakamura K,
Weinmaster G, Okano H, Nakafuku M: Role ofDeltex-1 as a transcriptional regulator downstreamof the Notch receptor. J Biol Chem 276: 45031-45040,
2001
23. Yamamoto S, Nagano M, Sugimori M, Kosako H,
Nakatomi H, Yamamoto N, Takebayashi H,
Nabeshima Y, Kitamura T, Weinmaster G, Nakamura
K, Nakafuku M: Transcription factor expression andNotch-dependent regulation of neural progenitors inthe adult rat spinal cord. J Neurosci 21: 9814-9823,
2001
24. Yamamoto S, Yamamoto N, Kitamura T, Nakamura
K, Nakafuku M: Proliferation of parenchymal neuralprogenitors in response to injury in the adult rat
spinal cord. Exp Neurol 172: 115-127, 2001
25. Ikeda T, Mabuchi A, Fukuda A, Hiraoka H,
Kawakami A, Yamamoto S, Machida H, Takatori Y,
Kawaguchi H, Nakamura K, Ikegawa S: Identificationof sequence polymorphisms in two sulfation-related
genes, PAPSS2 and SLC26A2, and an association
analysis with knee osteoarthritis. J Hum Genet 46:
538-543, 2001
26. Kawaguchi H, Nakamura K, Tabata Y, Ikada Y,
Aoyama I, Anzai J, Nakamura T, Hiyama Y, Tamura
M: Acceleration of fracture healing in non-humanprimates by fibroblast growth factor-2. J Clin
Endocrinol Metab 86: 875-880, 2001
27. Koshizuka Y, Ikegawa S, Sano M, Nakamura K,
Nakamura Y: Isolation of novel mouse genes associated with ectopic ossification by differential display
method using ttw, a mouse model for ectopic ossification. Cytogenet Cell Genet 94: 163-168, 2001
28. Seichi A, Takeshita K, Ohnishi I, Kawaguchi H,
Akune T, Anamizu Y, Kitagawa T, Nakamura K:
Long-term results of double-door laminoplasty forcervical stenotic myelopathy. Spine 26: 479-487,
2001
29. Ogata N, Shiraki M, Koshizuka Y, Nakamura K,
Kawaguchi H: A polymorphic variant at the Wernerhelicase (WRN) gene is associated with bone density,
but not spondylosis, in postmenopausal women. J
Bone Miner Metab19: 296-301, 2001
30. Nakajima S, Nakamura K, Masamura K, Sakuma I,
Dohi T: Three-dimensional medical imaging display
with computer-generated integral photography.
Comput Med Imaging Graph 25: 235-241, 2001
31. Nakajima S, Nakamura K, Masamura K, Sakuma I,
Dohi T: Development of a tracking method for image
analysis of time-lapse deformation of trabecular
structure in a human bone. Comput Med Imaging
Graph 25: 443-448, 2001
32. Koshizuka Y, Ikegawa S, Sano M, Nakamura K,
Nakamura Y: Isolation, characterization, and mapping of the mouse and human WDR8 genes, members of a novel WD-repeat gene family. Genomics 72:
252-259, 2001
33. Nishimura G, Manabe N, Kosaki K, Haga N, Ohashi
H, Nakamura K, Ikegawa S: Spondylar dysplasia in
Type X collagenopathy. Pediatr Radiol 31: 76-80,
2001
34. Akune T, Ogata N, Seichi A, Ohnishi I, Nakamura K,
Kawaguchi H: Insulin secretory response is positively
associated with the extent of ossification of the posterior longitudinal ligament of the spine. J Bone Joint
Surg Am 83-A: 1537-1544, 2001
35. Mabuchi A, Ikeda T, Fukuda A, Koshizuka Y,
Hiraoka H, Miyoshi K, Haga N, Kawaguchi H,
Kawakami A, Yamamoto S, Takatori Y, Nakamura K,
Ikegawa S: Identification of sequence polymor-

phisms of the COMP (cartilage oligomeric matrix
protein) gene and association study in osteoarthrosisof the knee and hip joints. J Hum Genet 46: 456-462,
2001
36. Mabuchi A, Haga N, Ikeda T, Manabe N, Ohashi H,
Takatori Y, Nakamura K, Ikegawa S: Novel mutationin exon 18 of the cartilage oligomeric matrix protein
gene causes a severe pseudoachondroplasia. Am J
Med Genet 104: 135-139, 2001
37. Fukui N, Nakajima K, Tashiro T, Oda H, Nakamura
K: Neutralization of fibroblast growth factor-2
reduces intraarticular adhesions. Clin Orthop 383:
250-258, 2001
38. Fukui N, Fukuda A, Kojima K, Nakajima K, Oda H,
Nakamura K: Suppression of fibrous adhesion byproteoglycan decorin. J Orthop Res 19: 456-462, 2001
39. Ikeda T, Mabuchi A, Fukuda A, Kawakami A,
Yamada R, Yamamoto S, Miyoshi K, Haga Y, Hiraoka
H, Takatori Y, Kawaguchi H, Nakamura K, Ikegawa
S: Association analysis of single nucleotide polymorphisms in cartilage-specific collagen genes with knee
and hip osteoarthritis in the Japanese population. J
Bone Miner Res 17: 1290-1296, 2002.
40. Yamamoto A, Miyazaki T, Kadono Y, Takayanagi H,
Miura T, Nishina H, Katada T, Wakabayashi K, Oda
H, Nakamura K, Tanaka S: Possible involvement of
IkB kinase 2 and MKK7 in osteoclastgenesis induced
by receptor activator of nuclear factor kB ligand. J
Bone Miner Res 17: 612-621, 2002
41. Takayanagi H, Kim S, Matsuo K, Suzuki H, Suzuki T,
Sato K, Yokochi T, Oda H, Nakamura K, Ida N,
Wagner EF, Taniguchi T: RANKL maintains bonehomeostasis through c-Fos-dependent induction ofinterferon-β. Nature 416: 744-749, 2002
42. Matsuda K, Yoshida K, Taya Y, Nakamura K,
Nakamura Y, Arakawa H: p53AIP1 regulates the
mitochondrial apoptotic pathway. Cancer Res 62:
2883-2889, 2002
43. Koshizuka Y, Kawaguchi H, Ogata N, Ikeda T,
Mabuchi A, Seichi A, Nakamura Y, Nakamura K,
Ikegawa S: Nucleotide pyrophosphatase gene polymorphism associated with ossification of the posterior longitudinal ligament of the spine. J Bone Miner
Res 17: 138-144, 2002
44. Shimoaka T, Ogasawara T, Yonamine A, Chikazu D,
Hirotaka H, Nakamura K, Itoh N, Kawaguchi H:
Regulation of osteoblast, chondrocyte, and osteoclast
functions by fibroblast growth factor (FGF)-18 incomparison with FGF-2 and FGF-10. J Biol Chem
277: 7493-7500, 2002
45. Ogata N, Matsumura Y, Shiraki M, Kawano K,
Koshizuka Y, Hosoi T, Nakamura K, Kuro-o M,
Kawaguchi H: Association of klotho gene polymorphism with bone density and spondylosis of thelumbar spine in postmenopausal women. Bone 31:
37-42, 2002
46. Ogata N, Koshizuka Y, Miura T, Iwasaki M, Hosoi T,
Shiraki M, Seichi A, Nakamura K, Kawaguchi H:
Association of bone metabolism regulatory factor
gene polymorphisms with susceptibility to ossifica-269

tion of the posterior longitudinal ligament of thespine and its severity. Spine. 27: 1765-1771, 2002
47. Kawano K, Ogata N, Chiano M, Molloy H, Kleyn P,
Spector TD, Uchida M, Hosoi T, Suzuki T, Orimo H,
Inoue S, Nabeshima Y, Nakamura K, Kuro-o M,
Kawaguchi H: Klotho gene polymorphisms associated with bone density of aged postmenopausal
women. J Bone Miner Res (in press)
48. Ikeda T, Junwei Z, Chano T, Mabuchi A, Fukuda A,
Kawaguchi H, Nakamura K, Ikegawa S: Identificationand characterization of the human long form of Sox5
(L-SOX5) gene. Gene (in press)
49. Kotani H, Kawaguchi H, Ueno S, Shimoaka T,
Iwasaka M, Ozawa H, Nakamura K, Hoshi K: Strong
static magnetic field stimulates bone formation to adefinite orientation in vitro and in vivo. J Bone Miner
Res (in press)
50. Akune T, Ogata N, Hoshi K, Kubota N, Terauchi Y,
Tobe K, Takagi H, Azuma Y, Kadowaki T, Nakamura
K, Kawaguchi H: Insulin receptor substrate-2 maintains predominance of anabolic function over catabolic function of osteoblasts. J Cell Biol (in press)

270

Department ofOphthalmology
Outline and Research Objectives
The Department of Ophthalmology University of Tokyo was established in 1889. Since
then, over a century, our department has played a crucial role as a pioneer in every aspects
of ophthalmology in Japan and delivered a lot of international leaders in both clinical andresearch fields of ophthalmology. The chairman Jujiro Komoto (1889-1922) introduced
Western ophthalmology into Japan and established the basics in Japanese ophthalmologic
society. The chairman Shinobu Ishihara (1922-1940) developed Ishihara test plate for color
blindness, which is still a world-wide golden standard as a screening examination for color
blindness. The chairman Yoshiharu Shoji (1940-1949) investigated on the pathogenesis of
cataract and the chairman Hagiwara (1915-1964) conducted basic and clinical research on
Behcet’s disease, the cornea, and the extra-ocular muscles. The chairman Shinichi Shikano(1964-1971) performed research on Behcet’s disease and autonomic nervous system in theeye. The chairman Saiichi Mishima (1971-1987) controlled comprehensive research on thephysiology of the cornea, ocular pharmacology, and surgical biology, and served as a chieforganizer of international congress of ophthalmology held in Kyoto in 1978. The chairman
Kanjiro Masuda (1987-1997) conducted research on the pathogenesis and therapy of uveitis.
The chairman Makoto Araie (1997-present) has organized thorough research on ocular pharmacology, ocular blood-flow, and clinical and basic aspects of glaucoma and cornea transplantation.

Faculties and Students:
Professor and Chair
Associate Professor

Makoto Araie, MD., Ph.D. (1997~)
Hidetoshi Kawashima, MD., Ph.D.
Goji Tomita, MD., Ph.D.
Lecturer
Yasuyuki Suzuki, MD., Ph.D.
Yasuhiro Tamaki, MD., Ph.D.
Satoru Kato, MD., Ph.D.
Miyuki Nagahara, MD., Ph.D.
Associate ...............................10
Postdoctoral Fellow ................1
Secretary .................................2

Past Researches and Major accomplishments
1) Evaluation of important parameters of ocularphysiology by means of in vivo fluorophotometry
or in vivo measurements of protein concentrationin the aqueous humor:
Effects of various anti-glaucoma eye drops on boththe aqueous flow rate and blood-aqueous-barrier(BAB) permeability were first and quantitatively determined in human eyes and anti−inflammatory effects
of topical NSAID were first quantitatively determinedin patients undergoing cataract surgery. Further, amethod to non-invasively monitor the aqueous production reducing effects of anti−glaucoma drugs inhuman eyes or toxic effects of intra-ocular irrigating

solution in animal eyes have been first developed.
The techniques thus establish were used in developing a new anti-glaucoma drug, which is now marketed.

2) Study of cornea:
Permeability to the cornea is the rate-limiting factor for intra-ocular penetration of topically instilled
drugs. A pre-existing method for in vivo determination of corneal endothelial (CE) permeability was
improved so that status of its harrier function can be
estimated more systematically. A new in-vitro
method for quantitative evaluation of CE permeability
was established, which was world-wide used for comparing safety of various intra-ocular irrigating solutions, and the lowest Ca 2+ concentration needed tomaintain the CE integrity was first determined.
Further, molecular biological features of corneal
endothelial cells were investigated and the above
studies are followed by the present investigation ofregenerative medicine of corneal graft for corneal
transplantation.
3) Study of ocular pharmacokinetics:
Using the 2-compartment computer model, ocularpharmacokinetics of topically instilled beta-blockers,
one of the most important anti-glaucoma drug, werestudied and binding of topically instilled beta-blockers
to uveal pigment and its clinical implication were first271

demonstrated. Trans-corneal and intravitreal kineticsof flourophores were studied to determine factors tofacilitate intra-ocular penetration of topically instilled
and systemically administered drugs.

4) Clinical study of glaucoma:
Glaucoma is the leading cause of blindness indeveloped countries and open angle glaucoma represents 75% of all glaucomas in Japan. Surgical techniques of glaucoma were improved by introducing
antimetabolites and basic features of adjunctive use ofanti metabolites investigated. By computer analysis ofvisual field results, IOP-dependent damages in thevisual field were elucidated, optimal sectorization oftest points determined and it was also found that IOP
is a risk factor for progression also in normal tension
glaucoma. And, effects of glaucoma surgery or calcium
antagonists on the visual field progression in normal
tension glaucoma were first qualitatively determined.
New image analysis techniques, psychophysical methods and new anti− glaucoma eye drops were evaluated and one anti−glaucoma eye drop could be successfully developed and marketed.
5) Basic study of glaucoma:
Clinical study of glaucoma demonstrated that IOP
is not the only factor contributing to progression ofglaucoma, which lead to experimental studies on neuroprotection. Conditioned medium from human amniotic epithelial cells was first found to have neurotrophic effects on cultured neurons including retinal
ganglion cells (RGCS) and interaction between retinal
glial cells or NO and isolated RGCs were determined.
Novel gene mutation in Japanese glaucoma patientswas found and MYOC, one of glaucoma genes, wascloned in the bovine eye.
6) Study of ocular blood flow:
Clinical studies of glaucoma revealed that local circulation is also involved in progression of glaucoma.
Apparatuses which can non-invasively estimate theblood flow in the iris, retina, choroids or optic nerve
head utilizing laser speckle phenomenon in livingeyes were constructed, establishing the laser speckle
method as an in-vivo, non-invasive peripheral bloodflow measurement method which can be applied inhuman subjects. Using this method, effects of calcium
antagonists, topical eye drops or surgical procedureson ocular circulation of various ocular tissues could befirst systemically studied. It was first found that theocular tissue most effectively affected is different
depending on the Ca2+-antagonist used (nicardipine
selectively affects choroidal circulation, lomerizine
ONH circulation etc). The important new finding
obtained by the ocular blood studies was, topicallyinstilled drugs can influence the circulation in the pos-272

terior parts of the eye through their direct penetration
to the posterior part of the eye, which have been considered to be impossible. This finding not only has
great clinical implication in ocular therapeutics, butalso prompted the current study to confirm that topically instilled drugs penetrate to the posterior retina,
choroids or retrobulbar tissues at pharmacologicalconcentrations.

Current Researches and Future Prospects.
1) Study of Cornea:
To cope with very limited supply of donor corneas
and allograft rejection in corneal transplantation, thereconstruction of artificial cornea has been attemptedusing the tissue engineering technology. A method to
culture human corneal endothelial cells (HCEC) has
been first established and, using cultured HCFCs andhuman corneal stroma the cornea could be successfully reconstructed. Thus reconstructed cornea has 7080% of cell density, pump function and barrier function of normal HCECs. The reconstructed cornea
transplanted to rabbit eye stayed transparent during
postoperative 6 months, showing transplanted HCEC
normally functioning. Animal corneal stroma as a carrier of cultured HCECs is being investigated. The
cornea reconstructed with porcine corneal stroma and
HCECs showed average cell density pump function of
about 60-70% of normal HCECs and bone marrowcells injected into rat anterior chamber were found totransform into CE-like cells, suggesting that self
immature cells can transform into CEs. Possibility ofutilizing cultured HCEC from fetus is now investigated. Antigen-specific T cell activation is a critical stepin the rejection of transplanted allografts. To activate
T cells, two kinds of signal are necessary: signal mediated by T cell receptor (TCR)/CD3 complex and the costimulatory signal by cell surface adhesion molecules
such as CD80/86 and ICAM-1. As novel therapies forrejection after corneal transplantation, the feasibilityof anti-CD80/86 antibodies is being investigated using
murine corneal transplantation model with a complete mismatch at major and minor histocompatibility. Clinical usage of reconstructed cornea, first in highrisk corneal transplantation cases, is planned in 5
years, to resolve very limited supply of donor corneas
in Japan. By incorporating gene technology, grafts
much better functioning with much less chance to be
rejected than the donor cornea may be constructed.
To reduce allograft rejection, therapy with anti-CD
80/86 antibodies and chemokine receptor antagonist
such as CCRI antagonist, which is also involved in
induction of rejection, may be also possible.

2) Study of ocular pharmacokinetics:
Our new finding suggesting penetration of topically instilled drug to the posterior parts of the eye at
pharmacological concentrations will be a breakthrough in medical treatment of diseases of posterior
parts of the eye such as the retinal, choroidal or optic
nerve disorders. Whole-head autoradiography and a
new method of isolation of posterior ocular tissues are
now being carried out and confirmed the above findings. Factors and routes contributing to the posterior
penetration of instilled drugs will be elucidated, andsystemically safe, local treatment of retinal, choroidal
or optic nerve disorders is to be developed.
3) Clinical study of glaucoma:
As one of the three chief investigators, Makoto
Araie conducted an epidemiological study of glaucoma
and related ocular diseases in Tajimi area and now isanalyzing the obtained results as a chief of data analysis committee. This is the first ophthalmological epidemiological study carried out in mongoloids and its
result will have a great impact in the field of ophthalmology. Several longitudinal, some of them randomized and prospective, studies are being conducted by
Makoto Araie as a chief investigator a) A 3−yearprospective, randomized，multi-centered and comparative study of nipradilol and timolol on the progression of normal tension glaucoma. The result of
this study will first elucidate if non-IOP dependentrisk factors of glaucoma can be treated by topicaldrugs or not, and if a positive result is obtained, it will
be a great break-through in the medical treatment ofglaucoma. b) The multi-centered, randomized, placebo-controlled study on the effects lomerizine, a Ca 2+blocker, on the retinal blood flow is the first to study
if retinal blood flow can be selectively increased bysystemically safe oral Ca2+-antagonis, lomerizine, andif a positive result is obtained, it will be a breakthrough in the medical treatment of retinal diseases.
Other longitudinal or cross-sectional studies now
ongoing includes disk bleeding study to investigate
whether myopic-type disc is associated less disc
bleeding but more progression of damage as compared with non-myopic type disc, advanced stage
glaucoma study where disability in daily life in thelate stage of the disease is prospectively questioned,
frequency doubling technology visual field study
where this new psychophysical test is prospectively
evaluated from view point of early detection of glaucoma progression and scanning laser ophthalmoscopy
study where morphometry of the optic disc inJapanese patients is quantitatively estimated on multicenter-based protocol, development of visual disability questionnaire designed to evaluate visual disability
in Japanese glaucoma patients who vertically read andwrite, analysis of filtering bleb characteristics after

trabeculectomy in Japanese patients to find contributing factors to the late stage bleb-related complications,the most vision-threatening complication of glaucoma
surgery. The result obtained from the above studies infuture will shed new light on the diagnosis, follow-up,
treatment and rehabilitation of glaucoma, the second
leading cause of blindness in Japan, of which prevalence in adults was found to be as high as 5% (unpublished result of the above mentioned Tajimi Study).

4) Basic study of glaucoma:
Makoto Araie is serving as a chairman of
Glaucoma Optic Neuropathy Study Group and established monkey glaucoma model is used to study how
glaucomatous damaging processes impair the optic
nerve head (ONH) structure, ONH circulation andneural functions (visual field, receptor field of neurons
in lateral geniculate body, etc) and how the potential
drugs can modify them, which is impossible to study
in humans. ONH is the tissue primarily involved inglaucomatous damage and ONH damage results inloss of retinal ganglion cells in retina. Using established system of culture of isolated retinal ganglion
cells and isolated Müller cells, factors, affecting survival of these cells, including therapeutic agents, are
now intensively studied. Mammalian retina is composed of seven major cell types, Müller glia, and six
neural cell types, retinal ganglion cell (RGC), bipolarcell, horizontal cell, amacrine cell, rod and cone photoreceptor cell. Unlike amphibian or fish, adult mammalian retina has been considered to be devoid ofstem or progenitor cells and that substantial regeneration dose not take place.
However, a recent study has unexpectedly demonstrated the presence of retinal stem cells in adultmice, suggesting the possibility that these cells can beutilized as a potential cell source of transplantation
therapy for damaged retinal cells including RGCs.
Animal model is pre-requisite for the realization of thetransplantation therapy, and rabbit is one of the mostsuitable animals because of its relatively large-sized
eyes, easiness in handling and reasonable price.
Method to isolate retinal stem cells from ciliary
epithelium using neurosphere-forming assay has been
established not only in mice but also rabbits andmonkeys. Ongoing studies address factors involved indifferentiation of retinal stem cells and how thesecells can be utilized as cell source of transplantation
therapy of retinal degenerative disorders including
glaucoma. In culturing isolated RGCs and retinal stem
cells, and in providing substrate for transplantation,
human amniotic epithelial cells and our experience onthem should be of considerable use.
5) Study of Ocular blood flow:
In addition to the laser speckle method that were273

already developed for non-invasive measurement of
peripheral blood flow in ocular tissues, an apparatus
for Color Doppler Imaging and that for Laser Doppler
Velocimetry have been recently introduced. The former measures the absolute blood velocity of vessels in
the orbit (diameter > 100µ) and the latter that of large
retinal vessels (diameter ≒100µ). Our laboratory is
the only one in the world that equipped with theabove 3 apparatuses and using 3 apparatuses, physiology, pathology and pharmacology of ocular circulationfrom feeding vessels to peripheral circulation can be
first systematically investigated. The subjects of the
on-going studies are: Measurement of peripheral
resistance in normal subjects with genetically highrisk for systemic hypertension; Change in autoregulatory capacity of retinal and ONH circulation withaging, development of glaucoma or development ofdiabetic retinopathy; and Assessment of hemodynamic parameters most correctly and sensitively reflecting
status of peripheral vascular bed. The results obtainedby above studies will shed light not only on further
understanding of pathophysiology of systemic hypertension and ocular disorders associated with circulatory abnormalities, but also lead to establishment of
hemodynamic parameters most sensitively reflecting
therapeutic effects of various drugs on retinal or ONH
disorders including glaucoma.

Research Grants
Grant-in-aid for scientific research from Ministryof Education, Culture, Sports, Science andTechnology:
1. Tenkai-Kenkyu (A)2, No.10357016, 1998-1999:| Chief
investigator, ¥19,500,000274

2. Kiban-Kenkyu (A)1, No. 11307036, 1999-2001: Chief
investigator, ¥41,010,000
3. Kiban-Kenkyu (B)2, No. 12557146, 2000-2001: Chief
investigator, ¥19,700,000
4. Kiban-Kenkyu (A)1, No. 14207069, 2002-2004: Chief
investigator, ¥49,998,000

Grant-in-aid for scientific research from Ministryof Health, Labour and Welfare. H-10-Sensory Organ-005, 1998-2000: Chief investigator, ¥73,500,000

Select Publications
1. Makoto ARAIE, Masahiro TAKASE, Yasuo SAKAI,
Yukihisa ISHII, Yukio YOKOYAMA & Masatoshi
KITAGAWA: Beta-adrenergic blockers: ocular penetration and binding to the uveal pigment. Japanese
Journal of Ophthalmology 26:248-263, 1982
2. Makoto ARAIE: A Reevaluation of corneal endothelial permeability to fluorescein. Experimental Eye
Research 41: 383-390, 1985
3. Makoto ARAIE: Barrier function of the corneal
endothelium and intraocular irrigating solutions.
Archives of Ophthalmology, 104;435-438, 1986
4. Makoto ARAIE & David M MAURICE: The rate ofdiffusion of fluorophores through the corneal epithelium and stroma. Experimental Eye Research 44: 7387, 1987
5. Chihiro SETO, Makoto ARAIE, Mitsuru SAWA, &Masahiro TAKASE: Human corneal endothelial permeability to fluorescein and fluorescein glucuronide.
Investigative Ophthalmology & Visual Science 28:1457-1463, 1987
6. Makoto ARAIE, Eiichi SHIRASAWA & Koji HIKITA:
Effects of oxidized glutathione on the barrier function of the corneal endothelium. InvestigativeOphthalmology & Visual Science 29: 1884-1887,
1988
7. Masami KONDO & Makoto ARAIE: Concentration
change of 5-fluorouracil in the external segment of
the eye after subconjunctival injection. Archives ofOphthalmology, 106: 1718-1721, 1988
8. Masami KONDO & Makoto ARAIE: Iontophoresis of
5-fluorouracil into the conjunctiva and sclera.
Investigative Ophthalmology & Visual Science 30:
583-585, 1989
9. Tetsuro OSHIKA & Makoto ARAIE: Time course ofchanges in aqueous protein concentration and flowrate after oral acetazolamide. Investigative
Ophthalmology & Visual Science 31: 527-534, 1990
10. Makoto ARAIE, Eiichi SHIRASAWA & TokieOHHASHI: Intraocular irrigating solutions and permeability of the blood-aqueous barrier. Archives ofOphthalmology, 108: 882-885, 1990
11. Makoto ARAIE, Koichi HAMANO, Shuichiro
EGUCHI & Shun MATSUMOTO: Effect of calcium
ion concentration of the permeability of the corneal
endothelium. Investigative Ophthalmology & Visual
Science 31:2191-2193, 1990
12. Makoto ARAIE & David M Maurice: The loss of fluorescein, fluorescein glucuronide and FITC-dextran
from the vitreous by the anterior and retinal pathways. Experimental Eye Research 52: 27-39, 1991
13. Mikiro MORI & Makoto ARAIE: A simple method ofdetermining the time course of timolol’s effects on
aqueous flow in humans. Archives ofOphthalmology 109: 1099-1103, 1991
14. Tetsuro OSHIKA, Makoto ARAIE, Tetsuya SUGIYAMA, Masayuki NAKAJIMA & Ikuo AZUMA: Effect ofbunazosin hydrochloride on intraocular pressure andaqueous humor dynamics in normotensive humaneyes. Archives of Ophthalmology 109: 1569-1574,
1991
15. Mikiro MORI, Makoto ARAIE, Masahiro SKURAI &Tetsuro OSHIKA: Effects of pilocarpine and tropicamide on blood-aqueous barrier permeability inman. Investigative Ophthalmology & Visual Science
33: 416-423, 1992
16. Yasuyuki SUZUKI, Makoto ARAIE & Yazuo OHASHI:
Sectorization of central 30∞ visual field in glaucoma.
Ophthalmology 100: 69-75, 1993
17. Satoshi KOYANO, Makoto ARAIE & Shuichiro
EGUCHI: Movement of fluorescein and its glucuronide across retinal pigment epithelium-choroid.
Investigative Ophthalmology & Visual Science 34:
531-583, 1993
18. Junkichi YAMAGAMI, Makoto ARAIE, Makoto
AIHARA & Seiichiro YAMAMOTO: Diurnal variation
in intraocular pressure of normal-tension glaucoma
eyes. Ophthalmology 100: 643-650, 1993
19. Makoto ARAIE, Junkichi YAMAGAMI & Yasuyuki
SUZUKI: Visual field defects in normal-and high-tension glaucomas. Ophthalmology 100: 1808-1814,
1993
20. Makoto ARAIE & Kiyoshi ISHII: Effects of apraclonidine on intraocular pressure and blood-aqueous bar-rier permeability after phacoemulsification andintraocular lens implantation. American Journal of
Ophthalmology 116: 67-71, 1993
21. Makoto ARAIE, Maki SEKINE, Yasuyuki SUZUKI &Nobuyuki KOSEKI: Factors contributing to the progression of visual field damage in eyes with normaltension glaucoma. Ophthalmology 101: 1440-1444,
1994
22. Yasuhiro TAMAKI, Makoto ARAIE, Eizo KAWAMOTO, Shuichiro EGUCHI & Hitoshi FUJII: Noncontact,
two-dimensional measurement of retinal microcirculation using laser speckle phenomenon. Investigative
Ophthalmology & Visual Science 35: 3825-3834,
1994
23. Yasuhiro TAMAKI, Makoto ARAIE, Eizo KAWAMOTO, Shuichiro EGUCHI & Hitoshi FUJITA: Non-contact, two-dimensional measurement of tissue circulation in choroid and optic nerve head using laser
speckle phenomenon. Experimental Eye Research 60:
373-384, 1995
24. Minoru KIMURA, Makoto ARAIE & Satoru KOYANO: Movement of carboxyfluorescein across retinal
pigment epithelium-choroid. Experimental Eye
Research 63: 51-56, 1996
25. Hiromu K MISHIMA, Kanjiro MASUDA, Yoshiaki
KITAZAWA, Ikuo AZUMA & Makoto ARAIE: A
comparison of latanoprost and timolol in primary
open-angle glaucoma and ocular hypertension. A 12week study. Archives of Ophthalmology 114: 929932, 1996
26. Nobuyuki KOSEKI, Makoto ARAIE, Shiroaki SHIRATO & Seiichiro YAMAMOTO: Effect of trabeculectomy on visual field performance in central 30∞ visual
field in progressive normal tension glaucoma.
Ophthalmology 104: 197-201;1997
27. Makoto ARAIE, Mariko KITAZAWA & Nobuyuki
KOSEKI : Intraocular pressure and central visual fieldof normal tension glaucoma. British Journal of
Ophthalmology 81:852-856,1997
28. Makoto ARAIE & Minoru KIMURA: Intraocular irrigating solutions and barrier function of retinal pigment epithelium. British Journal of Ophthalmology
81: 150-153;1997
29. Mikiko KANNO, Makoto ARAIE, Ken TOMITA &Kimio SAWANOBORI. Effects of topical nipradilol, aβ- blocking agent with α-blocking and nitroglycerin-like activities, on aqueous humor dynamics andfundus circulation. Investigative Ophthalmology &Visual Science 39:736-743,1998
30. Atsuo TOMIDOKORO, Makoto ARAIE, Yasuhiro
TAMAKI & Ken TOMITA. In vivo measurement of
iridial circulation using laser speckle phenomenon.
Investigative Ophthalmology & Visual Science
39:364-371,1998
31. Nobuyuki KOSEKI, Makoto ARAIE, Junkichi YAMAGAMI, Shiroaki SHIRATO & Seiichiro YAMAMOTO. Effect of oral brovincamine on visual field damage in normal tension glaucoma with low-normalpressure. Journal of Glaucoma 8:117-123,1999

275

32. Ken TOMITA, Makoto ARAIE, Yasuhiro TAMAKI,
Miyuki NAGAHARA & Tetuya SUGIYAMA. Effectsof nilvadipine, a calcium antagonist, on rabbit ocular
circulation and optic never head circulation in normal tension glaucoma subjects. Investigative
Ophthalmology & Visual Science. 40: 11441151,1999
33. Yasuhiro TAMAKI, Makoto ARAIE, Miyuki NAGAHARA & Ken TOMITA. Acute effects of cigarettesmoking on tissue circulation in human optic nerve
head and choroid-retina. Ophthalmolgy 106:564569,1999
34. Mikiko KANNO, Makoto ARAIE, Hiroshi KOIBUCHI& Kanjirou MASUDA. Effects of topical nipradilol, aβ-blocking agent with α-blocking and nitroglycerin-like activities, on intraocular pressure and aqueous dynamics in humans. British Journal of
Ophthalmology 84:293-299, 2000
35. Yuichi KAJI, Tomohiko USUI, Tetsuro OSHIKA,
Masao MATUBARA, Hidetoshi YAMASHITA,
Makoto ARAIE, Toshinori MURATA, Tetsuro
ISHIBASHI, Ryoji NAGAI, Seikoh HORIUCHI &Shiro AMANO. Advanced Glycation end products indiabetic corneas. Investigative Ophthalmology &Visual Science 41:362-368, 2000
36. Fumiko TANIGUCHI, Yasuyuki SUZUKI, Hiroki
KURIHARA, Yukiko KURIHARA, Hiroyoshi KASAI,
Shiroaki SHIRATO & Makoto ARAIE. Molecularcloning of the bovine MYOC and induction of its
expression in trabecular meshwork cells.
Investigative Ophthalmology & Visual Science
41:2070-2075, 2000
37. Kenji INOUE, Shunichiro KUBOTA, Tadahiko
TSURU, Makoto ARAIE & Yousuke SEYAMA.
Cholestanol induces apoptosis of corneal endothelial
and lens epithelial cells. Investigative
Ophthalmology & Visual Science 41:991-997, 2000
38. Tomohiko USUI, Shiro AMINO, Kaori SUZUKI,
Kazunori MIYATA, Makoto ARAIE, Paraskevi
HELDIN & Hidetoshi YAMASHITA. Expression andregulation of hyaluronan synthase in corneal
cndothelial cells. Investigative Ophthalmology &Visual Science 41:3261-3267, 2000
39. Saiko UCHIDA, Yukiko INANAGA, Makiko
KOBAYASHI, Shoei HURUKAWA, Makoto ARAIE &Norio SAKURAGAWA. Neurotrophic function of
conditioned medium from human amniotic epithelial
cells. J Neuroscience Research 62:585-590, 2000
40. Yasuhiro TAMAKI, Makoto ARAIE, Tomohiko
HASEGAWA & Miyuki NAGAHARA. Optic nerve
head circulation after intraocular pressure reduction
achieved by trabeculectomy. Ophthalmology 108:627-632, 2001
41. Ken MIZUNO, Takashi KOIDE, Mitsuo YOSHIMURA& Makoto ARAIE. Neuroprotective effect andintraocular penetration of nipradilol, a beta-blocker
with nitric oxide donative action. Investigative
Ophthalmology & Visual Science 42:688-694, 2001
42. Tomohiko USUI, Masumi HARA, Hiroaki SATOH,
Nobuo MORIYAMA, Humie KAGAYA, Shiro

276

AMANO, Tetsuro OSHIKA, Yasuo ISHII, Nobuhiro
IBARAKI, Chiaki HARA, Motoei KUNIMI, Eisei
Noiri, Kazuhisa TSUKAMOTO, Jun INATOMI,
Hayato KAWAKAMI, Hitoshi ENDOU, Takashi
IGARASHI, Astuo GOTO, Toshiro FUJITA, Makoto
ARAIE, George SEKI. Molecular basis of ocularabnormalities associated with proximal renal tubularacidosis. Journal of Clinical Investigation
2001;108:107-115
43. Yukihiko MASHIMA, Yasuyuki SUZUKI, Yuri
SERGEEV, Yuichiro OHTAKE, Tomihiko TANINO,
Itaru KIMURA, Hiroshi MIYATA, Makoto AIHARA,
Hidenobu TANIHARA, Masaru INATANI, Noriyuki
AZUMA, Takeshi IWATA, Makoto ARAIE. Novelcytochrome P4501B1(CYP1B1) gene mutations inJapanese patients with primary congenital glaucoma.
Investigative Ophthalmology & Visual Science
2001;42: 2211-2216
44. Kenji KASHIWAGI, Yoko IIZUKA, Makoto ARAIE,
Yasuyuki SUZUKI, Shigeo TSUKAHARA. Effects ofretinal glial cells on isolated rat retinal ganglion cells.
Investigative Ophthalmology & Visual Science
2001;42:2686-2694
45. Saiko UCHIDA, Yasuyuki SUZUKI, Makoto ARAIE,
Takashi SHIGEEDA, Takeshi HARA, Shiroaki
SHIRATO. Long-term follow-up of initial 5-fluorouracil trabeculectomy in primary open-angle glaucoma in Japanese patients. Journal of Glaucoma
2001;10:458-465
46. Kiyoshi ISHII, Atsuo TOMIDOKORO, Miyuki
NAGAHARA, Yasuhiro TAMAKI, Mikiko KANNO,
Yasuhiro FUKAYA, Makoto ARAIE. The effects of
topical latanoprost on optic nerve circulation in rabbits, monkeys and humans. Investigative
Ophthalmology & Visual Science 2001;42:2957-2963
47. Takashi SHIGEEDA, Atsuo TOMIDOKORO, Makoto
ARAIE, Nobuyuki KOSEKI, Seiichiro YAMAMOTO.
Long-term follow-up of visual field progression after
trabeculectomy in progressive normal-tension glaucoma. Ophthalmology 2002;109:766-770
48. Miyuki NAGAHARA, Atsuo TOMIDOKORO,
Sawako SANDO, Makoto ARAIE, Sumiyoshi
TANAKA. MD., Ph.D. MD., Ph.D. An apparatus forcolor Doppler imaging in seated subjects. American
Journal of Ophthalmology 2002;133:270-272
49. Hiroshi MATSUO, Atsuo TOMIDOKORO, Yasuyuki
SUZUKI, Shiroaki SHIRATO, Makoto ARAIE. Lateonset transconjunctival oozing and point leak of
aqueous humor from filtering bleb after trabeculectomy. American Journal of Ophthalmology 2002; 133:
456-462
50. Chihiro MAYAMA, Yasuyuki SUZUKI, Makoto
ARAIE, Kyoko ISHIDA, Akira TSUJI, Tetsuya
YAMAMOTO, Yoshiaki KITAZAWA, Shigeo
FUNAKI, Motohiro SHIRAKASHI, Haruki ABE,
Hidetoshi TSUKAMOTO, Koji OKADA, Hiromu K
MISHIMA. Myopia and advanced-stage open angle
glaucoma. Ophthalmology (in press)Department ofOtorhinolaryngology,
Head and Neck surgery
Outlines
The Department of Otorhinolaryngology was founded in 1899 by Prof. Waichiro Okada.
This is the first department of otorhinolaryngology of the medical school and the university
hospital in Japan. The first professor, Waichiro Okada (1902-1924), the 2nd, Taneji Masuda(1924-1946), the 3rd, Kotoji Satta, the 4th, Ichiro Kirikae (1947-1969), the 5th, Yasuo Sato
(1970-1980), the 6th, Yasuya Nomura (1980-1991), the 7th, Kimitaka Kaga (1992-)

Research Objectives
Our department covers all of otorhinolaryngological diseases and associated systemic
diseases, and has specialized clinical and basic research in middle ear and inner ear diseases,
peripheral and central deafness in infant and children, adult and elderly, facial paresis, vertigo and balance disorders, olfactory disorders, paranasal diseases, voice and speech disorders, cleft palate, taste and swallowing disorders, head & neck cancer, psychosomatic medicine and medical education.
1) Hearing research; Morphology and neurophysiology of the inner ear focusing on sensory
neural deafness: human temporal bone pathology, electron microscopic study in animalmodels with gene abnormality, clinical application of otoacoustic emissions and auditory
brainstem responses. Hair cell physiology of synaptic transmission.
2) Facial nerve research; Morphology of the facial nerves focusing on degeneration andregeneration.
3) Head and neck cancer research; Molecular biology and gene therapy of head and neckcancers.
4) Evoked potential research; Generators of auditory evoked potentials in the central auditory system.
5) Auditory perception and cognition research; normal subjects and patients with central
deafness and cochlear implant using event related potentials and MEG.
6) Voice research; Morphology and electrophysiology of the larynx and voice production.
7) Basic research on auditory brainstem implant.
8) Vestibular research; The oculomotor and balance system in the brainstem, cerebellum andcerebrum. Vestibular myogenic evoked potential and its origin and clinical application.
9) Taste research; Taste perception and disorders.
10) Olfactory research; Olfactory epithelium morphology and function of olfaction.
11) Language research; Development of hearing, speech and language in neonates andinfants.
12) Artificial organ research; Development of artificial sensory and motor organs.

Faculties and Students
Professor
Associate Professor
Lecturer

Kimitaka Kaga, MD,PhD (1992-)
Masashi Sugasawa, MD,
Tatsuya Yamasoba, MD
Toshihisa Murofushi, MD,PhD,
Naonobu Takeuchi, MD,PhD
Ken Ito, MD,PhD,
Kenichiro Ishio, MD
Ryuzaburo Higo, MD,PhD

Associate ...............................10
Postdoctoral Fellow ................2
Graduate Student....................6
Research Student ....................4
Secretary .................................3277

Past Research and Major Accomplishments

Current Research

Otology and neurotology
1. Development of new surgical procedure: total middle ear reconstruction and auricle, canal plasty andossicular reconstruction for congenital microtia
and atresia.
2. Comprehensive study on central auditory systemand central auditory disorders.
3. Demonstration of origins of auditory evoked
potentials in animal and human subjects: auditory
brainstem response and middle latency.
4. Introduction of gene therapy into inner ear diseases. 5. Demonstration of sound lateralization ofbone conduction hearing.
6. Invention of bilateral bone conduction hearing aid.
7. Discovery of a new hearing disorder of auditory
nerve disease.
8. Proposal of new critical age of language development.
9. Mechanism of perception and cognition of musicand environmental sounds.
10. Discovery of a new antibody of inner ear.

Otology and neurotology
I. Clinical research
1. Early operation of cochlear implant around two
years of age.
2. Music perception of children and adults withcochlear implant.
3. Sound lateralization perception and time-intensity
trading ability in normal subjects with absolute
pitch and patients with brain damage.
4. Development of super-low noise bone conduction
hearing aid.
5. Cooperative reconstruction surgery of auricle andcanal plasty for congenital microtia and atresia byplastic surgeons and otological surgeons.
6. New total middle ear reconstruction surgery for
infectious radicalized ears.
7. Implantation of bone anchored hearing aid forchildren with Treacher Collins syndrome.
8. Early discovery of congenital deafness in neonatesand early hearing aid fitting and education of
speech and language.
9. Contribution to double handicapped children withhearing problem.
10. Perception and cognition of cortical deafness.

Head and Neck Cancer:
1. Long term follow up study on different treatmentof maxillary cancer and proposal of the best treatment.
2. Identification of adeno virus in nasopharyngeal cancer.
3. Development of new chemotherapy of head andneck cancer.
4. Development of new surgical procedure for parapharyngeal tumor.
Rhinology:
Demonstration of developing and aging changesof olfactory neuroepithelium.
Laryngology:
Development of new artificial voice production
system for patients after total laryngectomy.
Bronchoesophagology
Development of electrophysiological swallowing
system.
Medical eduacation:
1. Introduction of clinical counselling with head andneck cancer patients for medical students in bedside learning.
2. Evaluation system of teachers in medical school
by medical students.

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II. Basic study
1. Development of polyimide multiple-site surface
microelectrode for epidural cortical recording ofhigh spatial resolution evoked-potentials.
2. Development of independent component analysis
of multiple-site auditory evoked potentials.
3. Molecular biology of inner ear hair cell differention.
4. Molecular biology of experimental noise induced
deafness.
5. Regeneration mechanism of inner ear hair cells inchick.
6. Development of gene therapy for experimental
inner ear diseases.
7. Origin of binaural interaction of hearing using
midline section of brainstem and localized lesionsof brainstem in animal models.
8. Plasticity of central auditory system.

Head and neck surgery
1. Development of a new procedure of skull base surgery.
2. Molecular biological study on hypopharyngeal
cancer.
3. Development of a new procedure of partial laryngectomy.

4. Quality of life between conservative and surgicaltreatment.
5. Development of navigation to support safer headand neck surgery.
Rhinology
1. Olfaction change of human subjects under environment of modern society.
2. A study on distribution of histamine receptor inhuman nasal mucosa.
3. Application of navigation system for endoscopic
nasal surgery.
Laryngology, Bronchoesophagology
1. Application of Botulinus toxin to treat spastic dysphonia.
2. A new procedure to treat hoarseness in elderly
patients inplanting fascia in the vocal cord.
3. Language development of children using tracheal
canula in the first year of life.
4. Comparative study of vocal cords in differential
species.
5. Measurements of pressure changes of swallowing
system in patients with dysphagia.
6. Surgical procedures to treat dysphagia.
Medical education
1. How to teach humanity in medical education.
2. Development of assessment of teachers activity in
lectures, seminars and bedside learning in medical
education.

Future Prospects
New artifial sensory and motor organs and genetherapy will be new directions checked by gene analysis of each cancer individually and treated by
1. Otology: After present cochlear implant for profound sensory deafness, we will challenge to create a new cochlear implant for music perception,
auditory brainstem implant for neural deafness
and super low noise bilateral bone conduction
hearing aids for conductive deafness.
2. Head and neck surgery: Head and neck cancer
treatment will be checked by gene analysis of eachcancer individually and treated by gene therapy
due to phylosophy based on tailored medicine.
3. Rhinology: Allergic rhinitis is incurable diseases
but will be challenged by new immunotherapy.
Sensory and neural anosmia is desperate to recover at present but will be challenged to innovate
artificial sensory device for implantation in olfactory bulb.
4. Laryngology: Voice is very important in communication. Loss of vocal cord result from total laryngectomy because of surgical cancer treatment.

Now we are challenging to innovate new computerized devices for providing the same voice fromthe oral cavity before surgery which uses voicesynthesis technology and orally fitted voice generating system.

Research Grants
1998 Hatamura Y, Ishii T, Kaga K: Advanced medical
tools for clinicians and development of fine instruments. Ministry of Welfare. 1998-2001, Total¥400,000,000.
1999 Kaga K, et al: Basic study on auditory brainstem
implant. Ministry of Education. 1999-2001, Total¥14,000,000.
2000 Kaga K, et al: Study on auditory space in higher
brain. Ministry of Education. 2000-2001, Total¥6,000,000.
2001 Kaga K: Identification of sound localization inhuman brain. Ministry of Education ¥2,100,000.
2002 Kaga K, et al: Neonatal universal screening andlong term follow up. Ministry of Welfare and Labor¥1,260,000.

Select Publications
1. Kaga K, Uebo K, Sakata H, Abe Y, Haebara H,
Kosakai M: Auditory brainstem responses and temporal bone and brainstem pathology in brainstem
death, with special reference to autolysis of red
blood cells. Acta Otolaryngol (Stockh) 115:183186,1995.
2. Kaga K, Tanaka Y: Auditory air and bone conduction
brainstem responses and damped rotation test foryoung children with bilateral congenital atresia ofthe ears. Int J Pediat Otorhinolaryngol 32:1321,1995.
3. Kaga K, Tamai F, Kitazumi E, Kodama K: Auditory
brainstem responses in children with Cornelia deLange syndrome. Int J Pediat Otorhinolaryngol
31:137-146,1995.
4. Kaga K, Nakamura M, Shinogami M, Tsuzuku T,
Yamada K, Shindo M: Auditory nerve disease of bothears revealed by auditory brainstem reponses, electrocochleography and otoacoustic emissions.
Scandinavian Audiol 25:233-238,1996.
5. Ismail Noorhassim, Kaga K: Pure tones audiometry
and auditory brainstem responses in noise-induced
deafness. American J Otolaryngol 17:31-35,1996.
6. Suzuki M, Kaga K: Effect of cisplastin on the negative
charge barrier in strial vessels of the guinea pig. A
transmission electron microscopic study using polyethyleneimine molecules. Eur Arch Otorhinolaryngol
253:351-355,1996.
7. Yamada K, Kaga K, Uno A, Shindo M: Sound lateralization in patients with lesions including the auditory cortex: comparison of interaural time difference(ITD) discrimination and interaural intensity difference (IID) discrimination. Hearing Research 101:173180,1996.

279

8. Yamasoba T, Oka Y, Tsukuda K, Nakamura M, Kaga
K: Auditory findings in patients with maternallyinherited diabetes and deafness harboring a point
mutation in the mitochondrial transfer RNAleu(UUR)
gene. Laryngoscope 106:49-53,1996.
9. Yamasoba T, Suzuki M, Kaga K: Influence of chronic
kanamycin administration on basement membraneanionic sites in the labyrinth. Hearing Research
102:116-124,1996.
10. Kaga K, Shindo M, Tanaka Y: Central auditory information processing in patients with bilateral auditory
cortex lesions. Acta Otolaryngol (Stockh) Suppl
532:77,82,1997.
11. Tanioka H, Kaga K, Zusho H, Araki T, Sasaki Y: MRof the endolymphatic duct and sac: findings in
Meniere disease. AJNR Am J Neuroradiol 18:4551,1997.
12. Suzuki M, Kaga K: Effect of cisplatin on the basement membrane anionic sites in the ampulla, macula, and strial vascularis of guinea pigs. Ann Otol
Rhinol Laryngol 106:971-975,1997.
13. Nakamura M, Yamasoba T, Kaga K: Changes in otoacoustic emissions in patients with idiopathic sudden
deafness. Audiology 36:121-135,1997.
14. Yamada K, Kaga K, Sakata H, Uno A, Tsuzuku T:
Auditory evoked potentials under total spinal anesthesia in rats. Ann Otorhinolaryngol 10871092,1997.
15. Suzuki Y, Kaga K, Sugiuchi Y, Ishii T, Suzuki J-I,
Takiguchi T: Sudden bilateral hearing loss due togastric carcinoma and its histological evidence. J
Laryngol Otol 111:1142-1146,1997.
16. Kojima Y, Kaga K, Shindo M, Hirose A:
Electromyographic examination of patients with unilateral cortical facial paralysis. Otolaryng Head Neck
Surg 117:S121-124,1997.
17. Kaga K, Uebo K, Sakata H, Suzuki J: Auditory andvestibular pathology in brainstem death revealed byauditory brainstem response. Acta Otolaryng(Stockh) Suppl 503:95-103,1993
18. Uno A, Kaga K, Tsuzuku T, Kuroki M: Middle-latency responses of awake and anesthetized Japanesemacaques. Audiology 32:302-307,1993.
19. Kim LS, Kaga K, Tsuzuku T, Uno A: Effects of primary auditory cortex lesions on middle latency
responses in awake cats. A.N.L. (Tokyo) 20:155165,1993.
20. Yamada K, Kaga K, Tsuzuku T, Uno A: Analysis ofauditory brain stem response with lidocaine injection into the cerebrospinal fluid in rats. Ann Otol
Rhinol Laryng 103:796-800,1994.
21. Yamada K, Kaga K, Suzuki J-I: Temporal bone pathology in patients without caloric response. Acta
Otolaryngol (Stockh) 114:586-594,1994.
22. Iwasaki S, Kaga K: Chronological changes of auditory
brainstem responses in Cockayne's syndrome. Int J
Ped Otolaryng 30:211-221,1994.
23. Kaga K, Uebo K, Sakata H, Abe Y, Haebara H,
Kosakai M: Auditory brainstem responses and temporal bone and brainstem pathology in brainstem

280

death, with special reference to autolysis of red
blood cells. Acta Otolaryngol (Stockh) 115:183186,1995.
24. Kaga K, Ichimura K, Kitazumi E, Kodama K, Tamai F:
Auditory brainstem respnses in infants an children
with anoxic brain damage due to near-suffocation ornear-drowning. Int J Pediatr Otorhinolaryngol
36:231-239,1996.
25. Kaga K, Nakamura M, Shinogami M, Tsuzuku T,
Yamada K, Shindo M: Auditory nerve disease of bothears revealed by auditory brainstem responses, electrocochleography and otoacoustic emissions.
Scandinavian Audiol 25:233-238,1996.
26. Iwasaki S, Kaga K, Yagi M, Kuroda M: Vestibular
findings syndrome. ORL 58:343-346,1996.
27. Kaga K, Shinoda Y, Suzuki J-I: Origin of auditory
brainstem responses in cats: Whole brainstem mapping, and a lesion and HRP study of the inferior colliculus. Acta Otolaryngol (Stockh) 117:197-201,1997.
28. Kaga K, Ichimura K: A preliminary report clinicaleffects of otic solution of ofloxacin in infantile
myringitis and chronic otitis media. Int J Pediaric
Otorhinolaryngol 42:199-205,1998.
29. Suzuki M, Yamasoba T, Kaga K: Development of theblood-labyrinth barrier in the rat. Hearing Research
116:107-112, 19989.
30. Suzuki M, Yamasoba T, Kaga K: Effects of chronic
administration of Kanamycin on the basement membrane anionic sites in the crista ampullaris of guinea
pigs. Laryngoscope 108:81-86,1998j.
31. Sakamoto M, Sugasawa M, Kaga K, Kamio T:
Average thresholds in the 8 to 20 kHz range as afunction of age. Scand Audiol 27:189-192,1998.
32. Kaga K, Ono M, Yakumaru K, Owada M, Mizutani T:
Brainstem pathology of infantile Gaucher's disease
with only wave I and II of auditory brainstem
response. J Laryngol Otol 112:1069-1073,1998.
33. Kaga K, Takeuchi N, Sakurai H: Facial nerve pathology of two patients with herpes simplex encephalitis.
A temporal bone study. Facial Nerve, pp 1-6, Proc
VIII International Symposium on the Facial Nerve.
Kugler Pub, The Netherlands, 1998.
34. Kaga K, Yamada K, Tsuzuku T, Uno A: The longterm effect of bilateral inferior colliculus ablation on
auditory brainstem response in awake cats. Acta
Otolaryngol (Stockh) 119:128-131m1999.
35. Kaga K: Vestibular compensation in infants and children with congenital and acquired vestibular loss inboth ears. Int J Pediatr Otorhinolaryngol 49:215224,1999.
36. Naito R, Murofushi T, Mizutani M, Kaga K: Auditory
brainstem responses, electrocochleograms, andcochlear microphonics in the myelin deficientmutant hamster 'bt'. Hearing Research 136:4448,1999.
37. Yamasoba T, Tsukuda K, Oka Y, Kobayashi T, Kaga
K: Cochlear histopathology associated with mitochondrial transfer RNALeu(UUR) gene mutation.
Neurology 52:1705-1707,1999.

38. Kaga K, Shindo M, Tanaka Y, Haebara H:
Neuropathology of auditory agnosia following bilateral temporal lobe lesions: A case study. Acta
Otolaryngol 120:259-262,2000.
39. Takegoshi H, Kaga K, Kikuchi S, Ito K:
Mandibulofacial dysostosis: CT evaluation of thetemporal bones for surgical risk assessment inpatients of bilateral aural atresia. Int J PediatricOtorhinolaryngol 54:33-40,2000.
40. Yamada K, Kaga K, Tsuzuku T, Uno A: Long-termchanges in middle latency response and evidence ofretrograde degeneration in the medial geniculate
body after auditory cortical ablation in cats. Acta
Otolaryngol 120:744-949,2000.
41. Itoh K, Yumoto M, Uno A, Kurauchi T, Kaga K:
Temporal stream of cortical representation for auditory spatial localization in human hemispheres.
Neruoscience Letters 292:215-219,2000.
42. Kaga K, Setou M, Nakamura M: Bone-conductedsound lateralization of interaural time difference andinteraural intensity difference in children and ayoung adult with lateral microtia and atresia of theears. Acta Otolaryngol 121:274-277,2001.
43. Setou M, Kurauchi T, Tsuzuki T, Kaga K: Binauralinteraction of bone-conducted auditory brainstem
responses. Acta Otolaryngol 121:486-489,2001.
44. Sheykholeslami K, Kaga K, Tsuzuku T, Mizutani M:
Electrophysiological measures of auditory function in
the neurofilament-deficient mutant quail (Quv).
Hearing Research 153:115-122,2001.
45. Kaga K, Sakurai H, Ogawa Y, Mizutani T, Toriyama
M: Morphological changes of vestibular ganglion
cells in human fetuses and in pediatric patients. Int J
Pediatric Otorhinolaryngol 60:11-20,2001.
46. Komura Y, Tamura R, Uwano T, Nishijo N, Kaga K,
Ono T: Retrospective and prospective coding for predicted reward in the sensory thalamus. Nature
142:546-549,2001.
47. Sheykholeslami K, Kermany MH, Kaga K: Frequency
sensitivity range of the saccule to bone-conducted
stimuli measured by vestibular evoked myogenic
potentials. Hearing Research 160:58-62,2001.
48. Kaga K, Yasui T, Yuge T: Auditory behaviors andauditory brainstem responses of infants with hypogenesis of cerebral hemispheres. Acta Otolaryngol
122:16-20,2002.
49. Kondo K, Sagara H, Hirosawa K, Kaga K, Matsushima
S, Mabuchi K, Uchimura H, Watanabe T: Hair cell
development in vivo and in vitro: analysis by using a
monoclonal antibody specific to hair cell in the chickinner ear. J Comp Neurol 445:176-198,2002.
50. Ishimoto S, Ho K, Sasaki T, Shinogami M, Kaga K:
Total middle ear reconstructive surgery for the radicalized ear. Otol Neurotol 23:262-266,2002.

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Department of
Rehabilitation Medicine
Outline and Research Objectives
The Department of Rehabilitation Medicine was established by Ministry of Education,
Culture, Sports, Science and Technology in April 2001. It is one of the newest fields in the
University of Tokyo Graduate School of Medicine. It belongs to the Sensory and Motor
System Medicine Course of the Surgical Sciences Study Major. Current staff is only one professor. We have accepted the graduate school student formally since fiscal year 2001 when
the Rehabilitation Medicine Field was installed in. Two students of the first year class ofgraduate school and two of the second year class started some rehabilitation research. An
activity of our department is at one with the Central Rehabilitation Service of the university
hospital. As for the staff of the rehabilitation service of the university hospital, there are one
lecturer, one associate and three part-time medical doctors. In addition, one temporary associate has accompanied in April 2002, and he takes charge of the day hospital in psychiatry
rehabilitation.
The Central Rehabilitation Service derives from the establishment of the exercise therapy room in 1963 in the central medical service department. It aimed at the development of
the medical rehabilitation in our country corresponding to needs of rehabilitation service
that had become clear in the health care of the 20th century middle.
In the university hospital, the rehabilitation section increased and maintained ahydrotherapy room and the occupational therapy room, etc., and it was renamed the rehabilitation center in 1966. Then, it became an independent section by changing the title as
the central rehabilitation service in 1970. Both were names by measures in the hospital. A
formal name of the section was a department of physical therapy. The professor employment was set up as a full-time medical director in 1984. However, a formal name of the section was still a physical therapy department.
Rehabilitation medicine is a comprehensive medical management of disability including
its diagnosis and treatment. It was born in the flow of the health care and medical service atmodern ages by which it came to value the enhancement of not only extending years ofpatient’s life but also adding life to years. In Declaration of Alma-Ata by World HealthOrganization (WHO) of 1978, it was described, “primary health care addresses the main
health problems in the community, providing promotive, preventive, and curative and rehabilitative services”. Regardless of rapid expansion of those needs, acknowledgment of medical rehabilitation was delayed in the frame of conventional diagnosis and treatment department. In our country, it was 1996 that the rehabilitation department was authorized as theformal clinical department in the health insurance system by the former Ministry of Healthand Welfare. On the other hand, the professor of the physical therapy department was positioned as an assistance instructor in the sensory and motor system medicine course by the
University of Tokyo Faculty of Medicine having shifted to the graduate school course system between fiscal year 1995 and fiscal year 1997.
In the university hospital, the name was changed from the physical therapy department
to the rehabilitation medicine department by the budget measures in fiscal year 2001. Now
we integrated related occupational categories, which had historically belonged to the orthopedic surgery department and the former physical medicine department, and started themaintenance of the department. The Central Rehabilitation Service includes 14 physicaltherapists (one person is a part-time) and 5 occupational therapists and 5 acupuncture therapists as a rehabilitation team staff other than current medical doctors.
Our mission is to provide exceptional rehabilitation for our patients through clinicalexcellence and compassionate care, all dedicated to helping patients achieve their maximum
functional capacity and highest quality of life. Another significant focal point of the
Department is research. The research objectives of the Department are to make clear configuration of disabilities and to develop measures to reduce the obstacles for patients’ independent living as well as to promote reintegration in their society.

282

Faculties and Students
Professor and Chair Eto, Fumio, MD, Ph. D (1998~)
Lecturer
Kodama, Yoshiaki, MD, Ph. D
Associate .................................2
Visiting Fellow........................1
Graduate student ....................4
Secretary .................................1 (part-time employment)

Past Research and Major Accomplishments
The rehabilitation medicine field had not been
admitted as an established course before reorganization of the graduate school course system. Therefore,
there is still no own laboratory space. There is yet nospace of doctor’s medical office and other staff’s duty
rooms in the university hospital though the physicaltherapy department has been engaged in the medical
service activity of rehabilitation since 1963. In ourconventional research activity, a clinical research thatadheres to a daily medical management in rehabilitation has been a major subject.
Research topics that have been executed so far are
the following content.
1. The influence of the trouble in patient’s daily life
due to various diseases is analyzed, and an appropriate treatment program is developed. It is
included to describe natural course of the disease
and the trouble clearly in life cycle of physicallyhandicapped person. A study to make clear a pattern of the disorder type change in cerebral palsy
by the development process from infancy was
done. Natural course of Werdnig-Hoffmann disease and congenital muscular dystrophy, whichwere a rare pediatric disease, was described, andthe indication of rehabilitation was clarified.
2. Proper management of the factor disturbing rehabilitation process is important in everyday medical
rehabilitation care. The pain management was a
typical problem, and the pathophysiological study
on shoulder-hand syndrome in poststroke patientswas done.
3. Most of activities in daily life depend on walking
and locomotion ability. Especially, a study about
evaluation method of gait abnormalities anddevelopment of specific treatment of an abnormal
gait is necessary for regain of useful mobility inphysically disabled. The effect of various shoe
insoles including arch support as apparatus fortreatments was clarified by the gait analysis system.
4. We have studied on development of rehabilitation
therapy to promote independent living of thepatients with higher brain dysfunction due tocerebrovascular disorders, traumatic brain injury,
Alzheimer’s disease and so on. Such a study

included the development of assessment tools toevaluate the effectiveness of the rehabilitativeintervention.
5. The ordering a bed rest is one of the treatment
measures. We have clarified the meaning of disuse
syndromes in the rehabilitation as the adverse
effect, which includes the deconditioning state.
Moreover, decreased activity in daily life after discharge from hospital may be a significant cause of
disuse syndromes, especially in the older people.
The research on risk factors of accidental fall has
revealed that incidence of fall is higher in theolder people with decreased daily activities.
6. The disuse syndromes and hypoactive daily life
may be serious matter in the poststroke patientsliving in the community. We have examined theinfluence of daily activity on changes in the physical fitness of people with poststroke hemiplegia.
Then, we have concluded that they can improvetheir physical fitness without formal supervised
training by simply increasing their daily activities.
7. We have developed the comprehensive assessment
tool of activities of daily living (ADL) for the elderly based on the hierarchical structure of the ADL.
In rehabilitation of the dementia patients the usefulness of ADL measurement has been revealed
for classification of the severity of dementia as
well as for evaluation of the effect of rehabilitation
program.
8. A basic research including animal experiment isalso necessary to improve the rehabilitation management. In collaboration with some outside laboratories we have studied a causative mechanism ofmuscle atrophy, joint contracture and bone atrophy, which are main components of disuse syndromes. In addition, we have examined the effectof physical agents in such a condition; for example, effects of dietary calcium depletion and repletion on tibial bone volume, influences of exercise
and mechanical loading on bone strength, effects
of electrical stimulation for reducing the degree of
joint contracture, and so on.

Current Research
The department’s research interests are directed
towards reduction of the major disabilities of thehandicapped persons, namely immobility, instability
and intellectual impairment. The approach is boththeoretical, by seeking a better understanding of thebaneful processes which produce disability in daily
life of the handicapped persons; and practical by the
introduction and evaluation of new methods of management. Research workers of variety of medical andnon-medical backgrounds shall work together onthese problems. Such a research policy is the future283

directionality, but the present state is in extension of
the past research activities.
Two or more equipment (force platform, video
operation analysis system, surface electromyography,
and ataxiameter, etc.) are usually combined for theevaluation of walking and motion disturbances. This
combined operation analysis system is arranged forthe synchronized determination. In addition, synchronous measurements such as the torque machines forthe muscular power measurement and the surface
electromyography are possible. We have been analyzing human locomotion activities, and also examining
the effects of shoes with various insert or heel wedges
for walking in normal subjects and physically
impaired persons. The present research addresses
examining the effects of heel elevation of shoes on
walking of the patient with Parkinson’s disease.
Moreover, a biomechanical change in the walking pattern after surgical operation on the ligaments of theknee has been analyzed with those devices.
The accomplishment of human activities such as
locomotion requires the functional integration of atleast three major organic systems; the bone, joint, andmuscular system that carries out various motions indaily activities; central and peripheral nervous systemthat controls musculoskeletal system organs; cardiopulmonary system that supplies oxygen and glucose to the former two system organs. Therefore, clinical research interests of rehabilitation medicine are in
the development of integrated physical exercise program, the improvement of cardiopulmonary management during rehabilitation therapy, and restorative
design for brain damage and nerve injuries. Now weare studying on the evaluation of higher brain dysfunction concerning everyday activities and also on
the physical fitness measurement by using theergometer for the development of an appropriate
rehabilitation program.
The clinical practice of rehabilitation is characterized in the team approach by multidisciplinary cooperation. Because two or more specialized professionals
work together in practice, the area of research neededis broad. There are a lot of necessary, but unsettledresearch topics for development and enhancement of
rehabilitation practice. However, the physicians andnon-medical staffs cannot afford to spend time in the
research work because the needs of rehabilitation
medical service nowadays increase rapidly in the university hospital. Our present research environment isunder a poor condition, and therefore, more cooperation with the outside laboratory or organization
including other faculty of the university may be recommended to accomplish the research plan in order
to investigate a problem thoroughly.

284

Future Prospects
Rehabilitation medicine is an important task of primary health care. Theories of disability in health practice and research are area of a new frontier. A researchon treatment and prevention of disability depends onthe concept and the hypothesis of disability. In addition to an experimental study, a narrative based study
in ethnography is important so that the subjective
problems like disability experiences may be discussed.
Then, not only a biological approach but also a sociological approach should promote it concurrently.
Our academic field has just started. We haverequested a laboratory to accomplish experimental
study, but it is not given yet. Since historical process
and the financial situation of a current university, it ispessimistic to set up the research environment on theuniversity campus. We should struggle with the matter connected directly with the activity of the university hospital, which is a training organization of a
medical doctor, though there are an enormous number of unsolved research topics. We have entrusted
the attending lecturer to the staff of outside institutions and the organization in order to support ourclinical and educational tasks. We are now arranging
the cooperation system with staffs of these external
institutions for research and development of the welfare equipments to facilitate the independence of thedisabled persons. This system may be of help to theresearch activity coherent to clinical practice in rehabilitation.
The physical therapist and the occupational therapist are new type of jobs in our country in the development process, and they are positioned as medical
service staffs in the hospital. It is necessary toincrease the chance that those co-medical professionals also participate in the research activity in order toactivate a clinical research.
The research of rehabilitation medicine is a new
field, internationally. There increases a number ofinquiries to hope for studying at our department fromseveral foreign researchers of other Asian nations.
Unfortunately, our present environment is insufficientto accept them. Nevertheless, one Chinese doctor and
one Mongolia doctor are working with us now. The
former has already completed the doctor degree thesis,
and the latter is about to start a study now. Professor
Eto was invited to Chinese Rehabilitation Medicine
Association Conference held in Beijing in China in
October 2001 and gave a lecture, entitled “the current
situation of rehabilitation medicine in Japan, from aviewpoint of medical education”. In addition, he lectured several topics on stroke rehabilitation at the university hospitals in Wuhan city and Shijiazhuang city
in China. International cooperation in the rehabilitation medicine research is the future matter.

Table 1. Representative subjects for research grants

Amount of
Fiscal year Resource
Subject (Author)
money (yen)The MEXT ScientificStudy on effect of electric stimulation
1998-1999 Research Fund, Basicin prevention of disuse changes of
1,500,000
Research (C)bone and soft tissues (Dr. Akai, M)
The MEXT ScientificStudy on effect of heel elevation for
2000-2002 Research Fund, Basic
Parkinsonian gait (Dr. Eto, F)
4,100,000
Research (C)The MEXT ScientificStudy on functional recovery and ADL
2000
Research Fund, Basicof stroke patient's with higher brain
1,900,000
Research (C)dysfunction (Dr. Ohtsuru, I)
The MEXT ScientificStudy on evaluation of joint lesion by
2000
Research Fund, Basicusing noninvasive measures such as
3,100,000
Research (C)vibration response (Dr. Akai M)
Ministry of Health, Labor
Study on functional recovery and ADL
and Welfare, Researchin higher brain dysfunction (Dr. Eto, F)
2001-2002 Consignment Expense for
1,600,000
Psychiatric and Neurological
Disease(Abbreviation, MEXT: Ministry of Education, Culture, Sports, Science and Technology)

Research Grants
The total of the competing research expense,
which the doctor researcher who belonged to theCentral Rehabilitation Service of the university hospital acquired, is 13,200,000 yen since 1998. Five representative research topics and the capital sources are as
follows in table 1.

Select Publications
1. Ueda S, Miura T, Eto F. Shoulder-Hand Syndrome as
an important complication of hemiplegia. J Jap Ass
Phys Med Balneol Climatol 38: 35-42, 1975.
2. Eto F, Yoshikawa M, Morimatsu M, Hirai S, Ueda S.
Shoulder-hand syndrome as a complication of hemiplegia. Jpn J Geriat 12: 245-251, 1975.
3. Eto F, Ueda S. Long-term change of motor patterns in
cerebral palsy. Jpn J Rehabil Med 14: 261-266, 1977.
4. Eto F, Yoshikawa M, Ueda S, Hirai S. Posthemiplegic
shoulder-hand syndrome, with special reference torelated cerebral localization. J Am Geriatr Soc 28: 1317, 1980.
5. Eto F. Complications due to prolonged or excessive
rest in rehabilitation medicine. J Clinical andExperimental Medicine (Igaku no ayumi) 116: 416422, 1981.
6. Eto F. Activities of daily living of the patients withdementia in the long-stay psychogeriatric wards. In:
Flax HJ, Matta AA, eds. New Frontiers that Influence
Disease and Rehabilitation, IRMA IV, Puerto Rico,
1983, pp.125-129.
7. Watamori T, Eto F. A longitudinal study of linguistic
and nonlinguistic functions in a patient with
Alzheimer's disease. Rinshou Seishin Igaku (Clin
Psychiatry) 12: 1155-1168, 1983.

8. Eto F, Harasawa M, Hirai S. Hand dexterity related toage: Pegboard test as an indicator of aging and brain
dysfunction. Jpn J Geriat 20: 405- 409, 1983.
9. Eto F, Yoshikawa M. A dose-response study ofDihydroergotoxine (DHET) Mesylate for cerebrovascular disturbances. In: Gaitz CM, Samorajaski Y, eds.
Aging 2000: Our Health Care Destiny, vol 1:
Biomedical Issues, Springer-Verlag, New York, 1985,
pp.421-431.
10. Ueda S, Hasegawa T, Ando K, Sakuma A, Kusunoki
T. Standardization of the finger function test andupper extremity ability test for hemiplegia. Jpn J
Rehabil Med 22: 143-160, 1985.
11. Okawa Y, Ueda S, Eto F, Kikuchi N. The analysis of
the course of motor impairment in Fukuyama typecongenital muscular dystrophy. Jpn J Rehabil Med
22: 197-202, 1985.
12. Okawa Y, Eto F, Ueda S. The analysis of the courseof disability in Werdnig-Hoffman disease (chronictype). Jpn J Rehabil Med 22: 115-120, 1986.
13. Eto F. Treatment of degenerative neurological diseases in the elderly: a rehabilitation approach. Jpn J
Geriat 23: 35-40, 1986.
14. Eto F. Rehabilitation of post-stroke patients with cardiac disease. J Clinical and Experimental Medicine(Igaku no ayumi) 139: 691-695, 1986.
15. Ueda S, Eto F, Okawa Y. Standardization of a functional test for hemiplegia for the evaluation of total
body motion. Scand J Rehabil Med 17 (Suppl): 163,1988.
16. Eguchi K, Eto F, Ohkawa Y, Ueda S. Factors affecting
return to work following stroke in youth and young
adults. Jpn J Stroke. 10: 124-126, 1988.
17. Eto F. Physical disability and dementia. Jpn J Rehabil
Med 25: 85-86, 1988.
18. Eto F. Rehabilitation of the elderly patients. In: Hsu
C-J, Orimo H, eds. Problems of the aged. Asia &285

Pacific Council for Science & Technology, Taipei,
1989, pp64-88.
19. Majima M, Ueda S. Major factor of the deteriorationof physical fitness in poststroke hemiplegics. Jpn J
Rehabil Med 27: 53-57, 1990.
20. Eto F. Activities of daily living and mental status inelderly patients with organic brain disorders. In:
O'Neill D ed. Carers, Professionals and Alzheimer's
Disease, John Libbey & Company, London, 1991,
pp323-327.
21. Eto F. Falls and immobilization: A vicious circle in
the elderly. In: Orimo H, Fukuchi Y, Kuramoto K,
Iriki M eds. New Horizons in Aging, Univ of Tokyo
Press, Tokyo, 1992, pp.280-281.
22. Eto F, Tanaka M, Mizoguchi T. Comprehensive activities of daily living (ADL) index for the elderly. In
Beregi E, Gergely IA, Rajczi K, eds.: Recent Advancesin Aging Science, vol.2, Monduzzi Editore, Bologna,
1993, pp.2183-2187.
23. Obana M, Eto F. Progressive pes equinovarus complicated with Parkinson's disease. J Clin Rehabil 2: 7880, 1993.
24. Kakurai S, Akai M. Clinical experiences with a convertible thermoplastic knee-ankle-foot orthosis forpost-stroke hemiplegic patients. Prosthetics andOrthotics International 20:191-194, 1996.
25. Eto F, Saotome I, Akiya N, Furuichi T. Early intervention of rehabilitation service for acute stroke
patients. Eur J Neurol 3(Suppl 2): 102, 1996.
26. Akai M, Shirasaki Y, Tateishi T. Electrical stimulation on joint contracture; an experiment in rat modelwith direct current. Arch Phys Med Rehabil 78:405409, 1997.
27. Eto F. Rehabilitation of memory. Jpn J Rehabil Med
34: 395-399, 1997.
28. Akai M, Usuba M, Maeshima T, Shirasaki Y, Yasuoka
S. Laser's effect on bone and cartilage change
induced by joint immobilization; An experimentwith animal model. Lasers in Surgery and Medicine
21:480-484, 1997.
29. Kodama Y, Nakayama K, Fuse H, Fukumoto S,
Kawahara H, Takahashi H, Kurokawa T, Sekiguchi C,
Nakamura T, Matsumoto T. Inhibition of bone
resorption by pamidronate cannot restore normal
gain in cortical bone mass and strength in tail-suspended rapidly growing rats. J Bone Miner Res 12:1058-1067, 1997.
30. Usuba M, Akai M, Shirasaki Y. Effect of Low-Level
Laser Therapy (LLLT) on viscoelasticity of the contracted knee joint; Comparison with whirlpool treatment in rats. Lasers in Surgery and Medicine 22:8185, 1998.
31. Yokota K, Itsukaichi K, Kakurai S, Asakura K.
Consideration of training for walker-assisted gait:
From the investigation of the load under walker's
tips. Bull Jpn Soc Prosthetics Orthotics 14: 76-81,
1998.
32. Itsukaichi K, Kaneko H, Kakurai S, Tazawa E.
Habilitation of the congenital limb deficient children.
Bull Jpn Soc Prosthetics Orthotics 14: 88-94, 1998.

286

33. Eto F, Saotome I, Furuichi T, Ogasawara M. Effectsof long-term use of benzodiazepines on gait andstanding balance in the elderly. Annals of New York
Academy of Sciences 860: 543-545, 1998.
34. Kodama Y, Takeuchi Y, Suzawa M, Fukumoto S,
Murayama H, Yamato H, Fujita T, Kurokawa T,
Matsumoto T. Reduced expression of interleukin-11
in bone marrow stromal cells of senescence-accelerated mice (SAMP6): relationship to osteopenia withenhanced adipogenesis. J Bone Miner Res 13: 13701377, 1998, 1998.
35. Miyake N, Yanagihara K, Sindo N. House work offemale stroke patients. Jpn J Rehabil Med 36: 644648, 1999.
36. Kodama Y, Dimai HP, Wergedal J, Sheng M, Malpe R,
Kutilek S, Beamer W, Donahue LR, Rosen C, Baylink
DJ, Farley J. Cortical tibial bone volume in two
strains of mice: effects of sciatic neurectomy andgenetic regulation of bone response to mechanicalloading. Bone 25: 183-190, 1999.
37. Linkhart TA, Linkhart SG, Kodama Y, Farley JR,
Dimai HP, Wright KR, Wergedal JE, Sheng M,
Beamer WG, Donahue LR, Rosen CJ, Baylink DJ.
Osteoclast formation in bone marrow cultures from
two inbred strains of mice with different bone densities. J Bone Miner Res 14: 39-46, 1999.
38. Eto F. Recent advances in geriatric rehabilitation. Jpn
J Geriat 36: 153-161, 1999.
39. Xu H, Akai M, Kakurai S, Yokota K, Kaneko H. Effectof shoe modifications on center of pressure and inshoe plantar pressures, Am J Phys Med Rehabil
78:516-524, 1999.
40. Fujitani J, Ishikawa T, Akai M, Kakurai S. Influenceof daily activity on changes in physical fitness forpeople with post-stroke hemiplegia, Am J Phys Med
Rehabil 78:540-544, 1999.
41. Akai M, Shirasaki Y, Tateishi T. The effect of electricity on joint contracture due to immobilization;
Direct current stimulation in rat experiment. In:
Electricity and Magnetism in Biology and Medicine.
Bersani F ed., Kluwer Academic/Plenum Publishers,
New York, 1999, p.883-886.
42. Honda F, Akai M, Itsukaichi K, Kakurai S, Tazawa E.
A report on progress of trainings and obtained activities about electronic prosthesis fitting for the congenital limbs deficient child. Sagyo Ryoho (J Jpn Ass
OT) 18: 478-487, 1999.
43. Nara A, Umishima M, Imase A, Mineshima T. Studyon the effects of pulp-to-pulp distance in measuring
the pinch strength - compare the young with theaged in a nursing home -. Sagyo Ryoho (J Jpn Ass
OT) 19: 338-345, 2000.
44. Hirose K, Eto F. Exercise training for internal disability in the elderly. J Clin Rehabil 9: 3 42-350, 2000.
45. Eto F, Sakata T. Influential factors on health-related
quality of life after cerebral vascular disease. Jpn J
Geriat 37: 554-560, 2000.
46. Kodama Y, Miyakoshi N, Linkhart TA, Wergedal J,
Srivastava A, Beamer W, Donahue LR, Rosen C,
Baylink DJ, Farley J. Effects of dietary calcium deple-

tion and repletion on dynamic determinants of tibial
bone volume in two inbred strains of mice. Bone. 27:
445-452, 2000.
47. Kodama Y, Umemura Y, Nagasawa S, Beamer WG,
Donahue LR, Rosen CR, Baylink DJ, Farley JR.
Exercise and mechanical loading increase periostealbone formation and whole bone strength in
C57BL/6J mice but not in C3H/Hej mice. Calcif TissueInt. 66: 298-306, 2000.
48. Eto F. Causes of falls in the elderly. Japan Medical
Association Journal 44: 299-305, 2001.
49. Ohtsuru I, Eto F, Wada N, Saotome I, Furuichi T.
New reliable and valid motor assessment scale forstroke patients: Hemiplegic motor behavior tests.
Geriatrics Gerontology Int 1: 45-51, 2001.
50. Kodama Y, Eto F. How to find out patients with ahigh risk for falls. J Clin Rehabil 10: 961-964, 2001.

287

Department ofAnesthesiology
Outline and Research Objectives
The department of Anesthesiology was established in1952. Our department has residents and eight researchers from China. We give lectures and provide clinical education for
postgraduate students and medical students.
We have pain clinic services in our department, which was established in 1962. It is provided for not only out-patients but also including patients in the ward of the other departments on a daily basis in all areas of painful diseases.
Anesthesia service including pre-and post-operative care is provided every day for elective and emergency surgical procedures. Recently, we have started acute pain service forpostoperative pain relief and palliative medicine for cancer pain relief and terminal care.

Research activities:
There are eight laboratories in our department, which are investigating clinical and basic
project of anesthesia research.
These themes of the research groups of our department are listed as bellow.

Perioperative
anesthesia
management for
surgical patient
respiration
Emergency medicine
circulation
General anesthesia

Intensive care medicine

New
deviceandtechniqueforanesthesia
andpain medicine

Muscle relaxation
Resusitology

Local anesthesia

Shock
Pain medicine

Perioperative
Pain physiology

Palliative medicine

QOL

behavior

Anesthesiology, basic and clinical respiration
・ Measurement of pulmonary vascular resistance of normal and pulmonary edema using
rabbit・ The effect of anesthetics upon pulmonary function
・ The model of neurogenic pulmonary edema of rabbit
・ Basic research of lipopolysaccharide which interact with apoprotein from the lung of
neonatal mouse
・ Basic research of the changes in transmission and distribution of gas with pulmonary
lesion・ Clinical assessment of pulse-oximeter288

circulation
・ Relationship between sympathetic nervous system and cardiac function
・ Clinical assessment of anti-hypertensive agents during anesthesia
・ Prostaglandins and tissue blood flow of the liver and kidney
pain mechanism (neurobiology, neuro-physiology, behavior investigation)
・ Relationship between noxious stimuli and catecholamines in the anesthetized rat
・ Physiology of pain and the mechanisms of analgesia using spinal rat
・ The effect of midazolam upon the spinal analgesical mechanisms
・ The effect of peripheral sensory nervous activities by acupuncture
・ Basic and clinical assessment of benzodiazepine antagonist
・ Basic and clinical assessment of enkepharinase inhibitor・ Analgesic mechanism of nociceptin in spinal rat
・ Assessment of bispectral index monitor
・ Clinical evaluation of drug challenge test
・ Analgesic mechanism of laser treatment
・ Analgesic mechanism of oriental medicine
・ Clinical assessment of acupuncturemuscle relaxation
・ Mechanism and the pharmacological analysis of non-depolarizing muscle relaxants inhuman
・ Clinical assessment of malignant hyperthermia
・ Basic and clinical assessment of non-depolarizing muscle relaxants
new devices and techniques for anesthesia
・ Mechanism of H-type epidural-spinal anesthesia
・ Development of skin surface anesthetics by lidocaine tape・ Clinical assessment of the induction by midazolam combined with barbiturates
・ Anesthetic method by low dose buprenorphine with inhalational anesthetics
・ The effect of epidural buprenorphine and morphine on post-operative pain control
・ Clinical usefulness of patient-controlled analgesia (PCA)・ Clinical assessment of stellate ganglion block・ Assessment of anesthetic machines
・ Development of device for intubation
mechanism of anesthesia
・ Mechanism of anesthetics from auditory brainstem response and Fourier analysis of
EEG・ Inhalational anesthetics and rate potential
・ Analgesic mechanisms of hyperventilation
・ The interaction of central analgesic and spinal analgesic mechanisms
shock・ Response to immunological system against the endotoxin in the blood
・ The effect of poly-enzyme inhibitor upon tissue microcirculation
・ Effects of anesthetics upon liver cell and Kupffer cell
QOL in surgical patients during perioperative period and chronic pain patients・ Perioperative QOL in case of surgical patient
・ QOL of donor of blood
・ QOL of intractable chronic pain patient such as post-herpetic neuralgia and low back
pain

289

Faculty and Students

Future Project

Professor and Chair Kazuo Hanaoka, MD, Ph., D. (1991~)
Associate Professors Hideko Arita, MD, Ph.,D.
Tomoki Nishiyama MD, Ph., D
Lecturer
Cyoku Yajima, MD, Ph., D
Yasuo Ide, MD, Ph.,D.
Masakazu Hayasida MD, Ph.,D.
Associate ...............................13
Postdoctoral Fellow ..............12
Graduate student ....................9
Research student.....................6
Secretary .................................3

We are seeking the method of safe anesthesia
including perioperative period. Also safe managementof pain is really necessary in 21 century, because thepopulation of aged people will be tremendously
increased. Therefore, the investigated area of above
mentioned will be continued. Palliative care medicine
will be important field also in the near future.
Therefore, we will investigate this field. Actually clinical investigation of palliative care medicine has started from this year as a field study.

Research Grants
Past Research and Major Accomplishments
In Anesthesiology, Respiration, Circulation, Pain,
Muscle relaxation, New devices and techniques foranesthesia, Mechanism of anesthesia, Shock andPerioperative QOL are mainly involved.
For each project, we are investigating both basically and clinically. The main purpose of these investigation is that the safety and quality of perioperative
anesthesia care are sought, and also the treatments ofpatients with intractable pain as chronic pain andpostoperative pain as acute pain are important field of
pain management.
We investigated the effect of Drug Challenge Test
(DCT) for chronic intractable pain patient and established the easy and accurate method of DCT. This
DCT were applied to many chronic patients and got
good results by the treatment of administered adequate drugs orally. Epidurascopy for chronic lumbar
pain are investigated and developed clinically.
Postoperative pain management are also investigated and established guideline for the method using
local anesthetics combined with narcotics via epidural
catheter to the epidural space. Patient Controlled
Analgesia (PCA) and pre-emptive analgesia are also
investigated.

Current Research
Respiration, circulation, pain, muscle relaxation,
new devices and techniques for anesthesia, mechanism of anesthesia, shock and perioperative QOL arebeing investigated. Each project is progressing well.
Especially new devices for epiduroscopy with usefulattachment such as manipulating device are being
developed.
We are making of the animal model of post herpetic neuralgia (PHN), which is very useful for the investigation of PHN. The number of patients with PHN
will increase because of large population of high aged
people in the near future. Therefore, this is very
important investigation.

290

1. 1998-2001 Research Grant (C2), #10671404 The
Involvement of NMDA receptor in the intrinsic
analgesia evoked by hyperventilation.
2. 1998-2001 Research project. Grant-in-aid for scientific research (B)#10470315 The effects of vasoactive drugs upon the spinal pain modulation.
3. 1999-2002 Research Grant (C2), #11671480
The effects of inhalational anesthetics and nitricoxide upon the activities of spinal dorsal hornneuron.
4. 2000-2002 Research Grant (C2), #12671453
Investigation of network by analgesic mechanismsof spinal cord
5. 2002-2004 Research Grant(C2), #14571422 The
development and clinical application of cheeper
and more accurate neuromuscular monitoring
devices now in use.

Select Publications
respiration
1. Kitamura T, Uchida K, Tanaka N, Tsuchiya T,
Watanabe J, Yamada Y, Hanaoaka K, Seymour JF,
Schoch OD, Doyle I, Inoue Y, Sakatani M, Kudoh S,
Azuma A, Nukiwa T, Tomita T, Katagiri M, Fujita A,
Kurashima A, Kanagasaki S, Nakata K:Serologicaldiagnosis of idiopathic pulmonary alveolar proteinosis. American Journal of Respiratory & Critical
Care Medicine 162:658-662, 2000
circulation
2. Nishiyama T, Hanaoaka K,: Nicardipine did not activate rennin-angiotensin- aldosterone system duringisoflurane or sevoflurane anesthesia. Canadian
Journal of Anaesthesia 47(12):1249-1252,2000
3. Nishiyama T, Hanaoaka K: Free hemoglobin concentrations in patients receiving massive blood transfusions during emergency surgery for trauma.
Canadian Journal of Anaesthesia 47(19): 881-885,
2000
4. Nishiyama T, Yokoyama T, Matsukawa T, Hanaoka
K: Continuous nicardipine infusion to control bloodpressure after evacuation of acute cerebral hemorrhage. Canadian Journal of Anaesthesia 47(12):11961201,2000
5. Orii R, Sugawara Y, Hayashida M, Yamada Y, Kubota
K, Takayama T, Harihara Y, Makuuchi M, Hanaoaka
K:Peri-operative blood lactate levels in recipients ofliving-related liver transplantation. Transplantation
69(10):2124-7, 2000
6. Takeda K, Sawamura S, Tamai H, Hagihara R,
Hanaoaka K: Reversible tricuspid valve obstruction
during removal of renal cell carcinoma with intracardiac tumor extension. Anesthesia & Analgesia
91:1137-38, 2000
7. R. Orii, Y. Sawamura, M. Hayashida, Y. Yamada, K
Chang, T. Takayama, M. Makuudhi and K.Hanaoka:
Effects of amrinone on ischaemia-reperfusion injury
in cirrhotic patient undergoing hepatectomy: a comparative study with prostaglandin E1. British Journalof Anaesthesia 85(3): 389-95, 2000
8. Orii R, Sugawara Y, Hayashida M, Uchida K, Yamada
Y,Takayama T., Makuuchi T., Hanaoka K: Lactate iscorrelated with the indocyanine green elimination
rate in liver resection for cirrhotic patients.
Anesthesia & Analgesia 92(4):1064-070,2001
9. Kazuo Hanaoka, Akiyoshi Namiki, Shiji Dohi,
Yoshihisa Koga, Osafumi Yuge, Yasushi Kayanuma,
Kazuyuki Hidaka, Tadashi Kusunoki; A dose-ranging
study of midazolam for postoperative sedation ofpatients: A randomized, double- blind, placebo-controlled trial. Crit Care Med. 30 (6): 1256-1260, 2002
10. Masakazu Hayashida, Mieko Chinzei,, Haruko
Fujiwara, Kyoko Komatsu, Hisako Usui, Kanji
Uchida, Toshiya Tomioka, Kazuo Hanaoka:
Bispectral Index as an Indicator of Cerebral Function
during Surgery Using Deep Hypotheremia andCirculatory Arrest. Cardiovascular Anaesthesia 6(1):9-13, 2002

pain mechanism (neurobiology, neuro-physiology,
behavior investigation)
11. Tomoki Nishiyama, Takeshi Yokoyama, Kazuo
Hanaoka: Midazolam improves postoperative epidural analgesia with continuous impression of local
anesthetics. Canadian Journal of Anaesthesia 45(6) :551-555, 1998
12. T. Nishiyama and K. Hanaoka: Effect of diluent volume on post-operative analgesia and sedation produced by epidurally administrated Midazolam.
European Journal of Anesthesiology 15: 275-279,
1998
13. T. Nishiyama, T. Matsukawa and K. Hanaoka:
Continuous epidural administration of Midazolam
and Bupivacaine for postoperative analgesia. Acta
Anaethesiolgica Scandinaviva 43 : 568-572, 1999
14. Tomoki Nishiyama, Takashi Matsukawa, Kazuo
Hanaoka: Acute Phase Histopathological Study ofSpinally Administered Midazolam in Cats.
Anesthesia & Analgesia. 89: 717-20, 1999
15. Hanaoaka k, Hayashida M,Arita H, Sumida T, Ide
Y:Management of Postoperative Pain. The

Management of Acute and Chronic Pain The Use of
the Tools of Trade: 133-137,2000
16. Hanaoka K, Meno A, Arita H, Tamai H, Orii R,
Sumida T: Drug Challenge Tests in Chronic PainManagement. The Management of Acute andChronic Pain TheUse of the Tools of Trade: 345349,2000
17. H. Sekiyama, J Utsumi, S G Shimada, H Nagase andMeno A, Arita H, Hanaoaka K:The Efficacy of Drug
Challenge Test (DCT) in the Treatment of Pain
Attributable to Brachial Plexus Avulsion,
Management of Pain. World Perspective:183-186,
2000
18. Nishiyama T, Hanaoaka K: The effects of epidural
bupivacaine, morphine, and their combination on
thermal nociception with different stimulus intensity
in rats. Anesthesia &Analgesia 91(13) :652-656, 2000
19. Arita H, Meno A, Zhang L, Hanaoka K: The CPT and
Drug Treatment for Brachial Plexus Avulsion,
Management of Pain. A World Prespective:115-8,
2000
20. Nishiyama T, Gyermek L, Lee C, Kawasaki –Yatsugi
S, Yamaguchi T, Hanaoaka K:The analgesic interaction between intrathecal clonidine and glutamate
receptor antagonists on thermal and formalininduced pain in rats. 92 Anesthesia & Analgesia (3):725-732,2001
21. Hanaoaka K: Drug Challenge Tests in Chronic Pain
Patients. The role of catecholamines and other neuromediates in neuropathic pain. Proceeding of theSatellite Symposium of the 9 th International
Catecholamines Symposium: 69- 81,2001
22. Toshiya Tomioka, Yutaka Awaya, Kenji Nihei andKazuo Hanaoka: Post- Herpetic neuralgia in a patientwith congenital insensitivity to pain and anhirosis
Journal of Anesthesia 16(1): 84-6,2002
23. S. Swamura, T. Tomioka, and K. Hanaoka: The
Importance of Tail temperature monitoring during
tail-flick test in Evaluating the antinociceptive actionof valatile anesthstics. Acta Anaesthesiologica
Scandinavica 46:451-4, 2002
24. Hanaoka K., Hayashida M., Arita H., Sumida T., Ide
Y.: How to Set up an Acut Pain Service. Japan
Perspective, Recent Views on Clinical Pain Edited
by Varrassi G. Monduzzi Editor, Bologna: 93-97,
2002
25. Min Dai, Toshinobu Sumida, Megumi Tagami, Yasuo
Ide, Masaki Nagasae, Hiroshi Sekiyama and Kazuo
Hanaoka: Suppressive effect of spinal dorsal-horn
neuronal activity by local spinal-cord cooling inreversed by naloxone in cats. Journal of Anesthesia
16: 211-15, 2002
26. Li WM., Wu GC., Arita H., Hanaoka K.: Acupuncture
stimulation inhibits somoato-renal sympathetic Aand C-reflexes in anesthetized rats. Acupuncture &Electro-Therapeutics Research 27(2): 119-27, 2002.

muscle relaxation
27. Hideto Oyamada, Keiko Oguchi, Naoto Saitoh,Toshiko Yamazawa, Kenzo Hirose, Yoko kawara,291

Kazunao Wakatsuki, Katsuji Oguchi, Megumi
Tagami, Kazuo Hanaoka, Makoto Endo,andMasamitu Iino: Novel Mutations in C-teminal channel Region of the Ryanodire Receptor in Malignant
Hypermia Patients. Jpn. J. Pharmacol.88:159-166,
2002

new devices and techniques for anesthesia
28. Tomoki Nishiyama, Takashi Matsukawa, Kazuo
Hanaoka: The Effect of Age and Gender on the
Optimal Premedication Dose of Intramuscular
Midazolam. Anesthesia & Analgesia 86: 1103-1108,
1998
29. T. Nishiyama, N. Sugai and K. Hanaoka: Inivitro
Changes in the Transparency and PH of Cerebro
Spinal Fluid Caused by Adding Midazolam. European
Journal of Anaesthesiology 15: 27-31, 1998
30. Tomoki Nishiyama, Hideto Nakayama, Kazuo
Hanaoka,: Sevoflurane or Thipotenal-Isoflurane forInduction and Laryngeal Mask Insertion?
Comparison by Side Effects; Hemodynamics, andSpectral analysis of Heart Rate Variability.
Anesthesia and Resuscitation 35(2) : 99-103, 1999
31. Takayuki Kitamura, Yoshitugu Yamada, Hong-Lin
Du, Kazuo Hanaoka: Efficiency of a New Fiberoptic
Stylet Scope in Tracheal Intubation. Anesthesiology
91(6) :1628-1632, 1999
32. Kitamura T, Yamada Y, Du HL, Hanaoka K: An efficient technique for tracheal intubation using theStylet Scope alone. Anesthesiology 92(4) :1210-1211,
2000
33. T. Kitamura, Y.Yamada, M. Chinzei, H.L.Du andK.Hanaoka: Attenuation of heamodynamic responses
to tracheal intubation by the Stylet Scope. British
Journal of Anaesthesia 86(2):275-277,2001
34. Kato,M. Sugiyama,Y. Orii,R. Hayashida,M. Kaneko,J.
Takayama,T. Hanaoka, K. Makuuchi,M : Lactate levels in cirrhotic patients undergoing liver resection.
Hepato-Gestroentreology 48 (40):1106-9,2001
35. Toshiya Tomioka, Yutaka Awaya, Kenji Nihei,
Hiroshi Sekiyama, Shigehito Sawamura, Kazuo
Hanaoka : Anesthesia for Patients with Congenital
Insentivity to Pain and Anhidrosis; A Questionnarire
Study in Japan. Anesthesia & Analgesia 94:271274,2002
36. Nobuhide Kin, Masakazu Hayasida, Kyung-ho
Chang, Kanji Uchida, Kazuo Hanaoka: External manual compression of the abdominal aorta to control
hemorrhage from a ruptured aneurysm. Journal of
Anesthesia 16: 164-6, 2002
37. Tomoki Nishiyama,, Takashi Mastukawa, Kazuo
Hanaoka: Effects of adding Midazolam on the
Postoperative Epidural Analgesia with Two Different
Doses of Bupivacaine. Journal of Clinical Anesthesia
14: 92-7, 2002
38. Nishiyama T., Misawa K., Yokoyama T., Hanaoka K.:
Effects of combining Midazolam and Barbiturate on
the response toracheal intubation : changes in autonomic nervous system. Journal of Clinical Anesthesia
14(5):344, 2002

292

39. Nishiyama T., Hanaoka K., Ochiai Y.: The medianapproach to transsacral epidural block. Anesthesia &Analgesia 95(4): 1067-70, 2002
40. Tomoki Nishiyama, Takashi Mastukawa, Takeshi
Yokoyama, Kazuo Hanaoka: Rapid Inhalation
Induction with 7% Sevoflurane Combined with
Intravenous Midazolam.Journal of Clinical
Anesthesia 14:290-295, 2002

mechanism of anesthesia
41. Sawamura S, Kingery WA, Davies MF, Agashe G,
Clark DJ, Kobilka BK, Hashimoto T, Maze M:
Antinociceptive action of nitrous oxide is mediatedby stimulation of noradrenergic neurons in the brainstem and activation of alpha 2B adrenoceptors.
Journal of Neuroscience 20:9242-9251,2000
shock
42. T. Nishiyama, T. Yokoyama and K. Hanaoka: Effects
of sevoflurane and isoflurane anesthesia on arterial
ketone body ratio and liver function. Acta
Anaesthesiolgica Scandinavica 43: 347-351, 1999
43. Nishiyama T, Hanaoaka K: A traumatic asphyxia in achild. Canadian Journal of Anaesthesia 47:10991102, 2000
44. Nishiyama T, Matsukawa T, Hanaoka K: Is proteaseinhibitor a choice for the treatment of pre-or mild
disseminated intravascular coagulation? Critical Care
Medicine 28:1419-22, 2000
45. Takayuki Kitamura, Yoshitsugu Yamada, Yoshifumi
Beck, Sae Asai, Hong-Lin Du and Kazuo Hanaoka:
Postoperative left recurrent laryngeal nerve palsy
possibly caused by coincidental swelling of themetastatic mediastinal lymphonode. Journal of
Anesthesia 14(4):216-7,2000
46. Nishiyama T, Hanaoka K: Hemolysis in stored red
blood cell concentrates: modulation by haptoglobin
or ulinastatin, a protease inhibitor. Crit Care Med. 29(10):1978-82,2001
47. Nishiyama T, Hanaoka K: Do the effects of a protease inhibitor, ulinastatin, on elastase release byblood transfusion depend on interleukin 6 ? Crit CareMed. 29(11): 2106-2110,2001
48. Nishiyama T, Hanaoka K: Propofol-induced bronchoconstriction: two case reports. Anesthesia &Analgesia 93 (3): 645-646,2001
QOL pre-operation
49. Mina Nishimori, Akiko Takeoka, Arinobu
Tojyo,Yasuo Nakano, Yoshitsugu Yamada and Kazuo
Hanaoka: Self-reported recovery time of daily activity
after bone marrow harvesting from healthy doners.
Journal of Anesthesia 15:1-5,2001
50. Mina Nishimori, Nelly Moerman, Shunichi
Fukuhara, F. S. A. M. Van Dam, M.J. Muller, Kazuo
Hanaoka & Yoshitsugu Yamada: Translation and validation of the Amsterdam preoperative anxiety andinformation scale (APAIS) for use in Japan. Quality ofLife Research 11: 361-4, 2002

Department of Emergency
and Critical Care Medicine
OutlineDepartment of Emergency and Critical Care Medicine was established in 1965, as the
emergency service within the Central Clinical Service Facilities of the University of Tokyo
Hospital and at the same time as the intensive care service for in-hospital patients, it
became a tertiary emergency care and critical care center in the metropolitan Tokyo and also
became the principal teaching facility of the University of Tokyo. It is a designated Level I
Trauma Center, and also the home of one of the newest Life Flight aeromedical services in
the country.
The Emergency Center sees approximately 15000 patients per year. It contains major
trauma and cardiac resuscitation rooms complete with STAT X-ray and full monitoring andresuscitation equipment. There are 9 treatment spaces including space for orthopedic, gynecology, and optho-ENT evaluation. X-ray, rapid spiral CT, ultrasound, angiography and
STAT Lab are located adjacent to the Emergency Center.
The Critical Care Center contains adult intensive care unit (ICU) of 8 beds, cardiac careunit (CCU) of 6 beds, surgical high care unit (SHCU) of 36 beds, medical high care unit
(MHCU) of 15 beds, pediatric intensive care unit (PICU) of 6 beds, and neonatal intensive
care unit (NICU) of 6 beds.
The Emergency Center and Critical Care Center sees an excellent mix of multiple traumas, high-acuity medical, surgical, pediatric, and gynecological patients. The Life Flight
service provides another opportunity for exposure to critically ill patients. Consult services
are available from each of the clinical departments of the Hospital.

Faculties and Students
Professor and Chairman Naoki YAHAGI, M.D., Ph.D.
Associates ...............................9
Staff.........................................3
Research student.....................1
Secretaries ...............................3

Past Research and Major Accomplishments• Study of the effect of anesthetics on the thresholdfor transpulmonary passage of venous air emboli.
(Resuscitation 13: 81-86, 1986, Anesthesiology 67:
905-909, 1987, Anesth Analg 75: 720-723, 1992) .
We found that the threshold for transpulmonary passage of venous air was significantly
lower during halothane anesthesia (0.01-0.05
ml/kg), as compared with pentobarbital (1.0ml/kg), fentanyl (0.5 ml/kg), or ketamine (0.35
ml/kg) anesthesia.
• Study on the effect of catecholamine on regional
cerebral blood flow in dogs (Masui 36: 1176-1180,
1986)
Development of Hemoperfusion System (HF)
1) For the patients of cardiac surgery
2) For the regulation of water balance in the patientsof renal insufficiency
3) For the expansion of the indication of bloodlesspriming for cardiopulmonary bypass by hemoconcentration using HF.

The usage of HF during cardiopulmonary bypass in NCVC

1985

1986

1987

1988

1989

1990

Total

644

623

593

573

586

594

HF

13

78

115

84

120

180

HF in adults
(%)

13
2.0

56
9.0

83
12.0

44
7.8

67
11.6

97
16.3

HF in infants
(%)

0
0.0

22
3.5

32
5.4

40
7.0

52
8.9

83
14.0

293

• Development of HF pump system for the perioperative management of cardiac surgical patients suffering from renal insufficiency and deteriorated
cardiac function. (Ube-Junken; 1984)• Induction of hypothermia for the management of
the patient who undertaken open heart surgery
complicated by acute cerebral infarction (Kyukyuigaku 18:917-921, 1994) (1984).
• We elucidated the structural parts involved in activation and inactivation of Na channel by bioengineering and electrophysiology. (FEBS Letters 228:
195-200, 1987, Nature 339: 597-603, 1989) .
• Effect of low molecular weight dextran for ARDS
induced by oleic acid (Am J Emerg Med18: 180183, 2000)(1988).
• Yahagi cared 5300 cases of critically ill patients
including cardiac surgical patients, severely illpatients of all categories in NCVC, and emergency
cases. Yahagi developed a handy bronchofiberscope. (Easyscope:FiberTech, 21000BZZ00571000,1998). Yahagi engaged in the preparation of a manual for cardiac transplantation, and established
The International Society of Medical Gas in 1995.
• Induction of mild hypothermia for the management of severe cardiac insufficiency (Anesth Analg
79: 581-582, 1994 , J Clin Anesth 10: 120-125,
1998) (1991)

Inhaled nitric oxide (NO)1) Indication and the 2-4years follow up of the cases(Artif Organs 21: 17-20, 1997, Artif Organs 21: 8384, 1997, Artif Organs 22: 886-891, 1998, Artif
Organs 23: 169-174, 1999)

2) Fontan type operation (Ann Thorac Surg 57: 13711372, 1994)
3) LVAS (Artif Organs 19: 557-558, 1995, Ann Thorac
Surg 65: 345, 1998)• Helium inhalation (Anesth Analg 80: 1042-1045,
1995, Artif Organs 21: 24-27, 1997) We demonstrated the beneficial effect of helium/oxygen to
improve oxygenation of the postcardiac surgicalpatients who showed impairment of oxygenation
without physiological findings and with normal
chest radiographs despite having a positive end
expiratory pressure of up to 10 cmH2O.

The effect of helium-oxygen on oxygenation and lung parameters
Evaluation of Airway
1) Effect of change of position (Ann Thorac Surg min-90 minbaseline
He 90 min67: 894, 1999)
56*
PaO2/FiO2
117±45113±39174±56*
2) Ultrasound detection of diaphragmatic paraly- ±5* Qs/Qt (%)
27±7
29±6
19±5*
sis after cardiac operations (Ann Thorac Surg 18*
Cdyn (ml/cm H2O) 60±18
60±18
65±18*
65: 1841, 1998)±4
PIP (cm H2O)
25±4
25±3
25±4
3) Double lumen endobronchial tube Broncho- *: p<0.05 vs baseline values.
Cath II (Anesthesiology 81: 781-782, 1994)
4) The cause for needing airway maneuvers to • Autotriggering caused by cardiogenic oscillation
maintain a patent airway during the use ofduring flow-triggered mechanical ventilation (Crit
Care Med 28: 402-407, 2000)
cuffed oropharyngeal airway (COPA) using
endoscope, and the difficulty of maintaining • Clinical use of electrolyzed water (Artif Organs 21:
39-42,1997, Artif Organs 24:984-987, 2000).
patent airway during COPA use might by collapse of the larynx at the level of the hyoid • Elucidation of peripheral neural regulation of heart(Brain Res 761: 329-332, 1997, J Autonom Nerv
bone. (Resuscitation in press)
5) Diagnosis of airway obstruction (Anesth Analg
System 68: 43-48, 1998, Brain Res 794: 146-150,
76: 207, 1993, Anaesthesia 50: 91-92, 1995,
199, Brain Research 864: 157-161, 2000)
Anesth Analg 85: 1180-1181, 1997)
6) Bronchial lavage ofthe infant using alaryngeal mask airway ( Anaesthesia
49: 450, 1994)( Anaesthesia 49:
450, 1994)
7) Development of ahandy-type bronchofiberscope294

Current Research and Future Prospects
We are now concentrating to develop “the Vital
Care Network System” which is to manage the great
number of high risk people continually. The system iscomposed of wearable monitor, wrist watch type PHS
which is to amplify the signal, the information center
to receive and manage the vital sign signals and to callon hospitals for aid. (The Next Generation Software
Project of Information-technology Promotion Agency,
Japan. “Vital Care Network System”. 2002)

Research Grants
1. The Next Generation Software Project ofInformation-technology Promotion Agency, Japan.
“Vital Care Network System”. 2002. ¥128,000,000
2. JSPS Grants-in-aid for Scientific Research “The basic
research of electrolyzed water for clinical use”.
12470316 Basic Research (B)(2) 20002001.¥6,200,000
3. JSPS Grants-in-aid for Scientific Research “Intratissue
blood vessels detection using infrared for the aid of
minimum invasive surgery”. 13878179 Early Stage
Research. 2001-2002. ¥1,300,000
4. Grants-in-aid for Innovative Project of IndustryUniversity-Government Cooperation in Ministry of

Education, Culture, Sports, Science and Technology“The research for practical use of medical integral
videography” 2000-2003. ¥130,000,000
5. Tokyo Metropolis Grants-in-aid for IndustryUniversity-Government “Electrolyzed water for thebacterial wound healing” April 1999~Feb 2000.
¥6,569,000

Select Publications
1. Kitagawa H, Yamazaki T, Akiyama T, Yahagi N,
Kawada T, Mori H, Sunagawa K: Modulatory effects
of ketamine on catecholamine efflux from in vivo
cardiac sympathetic nerve ending in cats. Neurosci
Lett 24: 232-236; 2002
2. Kono M, Yahagi N, Kitahara M, Fujiwara Y, Sha M,
Ohmura A: Cardiac arrest associated with use of anargon beam coagulator during laparoscopic cholecyctectomy. Br J Anaesth 87: 644-646, 2001
3. Yahagi N, Kono M, Kitahara M, Fujiwara Y, Asakawa
Y, Katagiri J, Sha M, Ohmura A, Murakami A,
Takamoto S: Cause of airway obstruction during
cuffed oropharyngeal airway. Resuscitation 48: 275278, 2001
4. Yahagi N, Kono M, Kitahara M, Ohmura A, Sumita
O, Hashimoto T, Hori K, Ning-Juan C, Woodson P,
Kubota S, Murakami A, Takamoto S: Effect of electrolyzed waters on wound healing. Artif Organs 24:
984-987, 2000
5. Yahagi N, Matsui J, Matsui S, Amakata Y, Kumon K,
Ueda-Ishibashi H. Low molecular weight dextran
attenuates effects of oleic acid-induced lung injury inrats. Am J Emerg Med 18: 180-183, 2000
6. Yahagi N, Yamazaki T, Akiyama T: Either
desipramine or TMB-8 suppresses cyanide on
induced norepinephrine efflux from in vivo cardiac
sympathetic nerves of cats. Brain Research 864: 157161, 2000
7. Imanaka H, Nishimura M, Takeuchi M, Yahagi N,
Kumon K: Autotriggering due to cardiogenic oscillation during flow-triggering mechanical ventilation.
Crit Care Med 28: 402-407, 2000
8. Murakami A, Kaneko Y, Imanaka K, Takamoto S,
Yahagi N: Easy aortic cannulation: a tranxyphoidal
approach. Artif Organs 24: 156-157, 2000
9. Yahagi N, Shimizu R, Kono M, Kitahara M, Katagiri J,
Sha M, Ohmura A: Retractor to facilitate fiberscopeaided tracheal intubation. Resuscitation 41: 283-284,
1999
10. Kumon K, Yahagi N, Imanaka H, Takeuchi M,
Miyano H, Ohashi Y. Nitric oxide inhalation as achemical assist for the circulation in patients after
cardiac surgery. Artif Organs 23: 169-174, 1999
11. Yahagi N, Tanigami H, Watanabe Y, Kumon K:
Semiprone position relieved airway obstruction
resulting from dilated pulmonary artery. Ann Thorac
Surg 67: 894, 1999
12. Yahagi N, Kumon K, Tanigami H, Watanabe Y,
Haruna M, Hayashi H, Takamoto S. Cardiac surgery

295and inhaled nitric oxide: Indication and follow up.
Artif Organs 22: 886-891, 1998
13. Yahagi N, Kumon K, Watanabe Y, Tanigami H,
Haruna M, Hayashi H, Imanaka H, Takeuchi M,
Ohashi Y, Takamoto S: Value of mild hypothermia inpatients who have severe circulatory insufficiency
despite the use of IABP. J Clin Anesth 10: 120-125,
1998
14. Yahagi N, Akiyama T, Yamazaki T: Effect of omegaconotoxin GVIA on cardiac sympathetic nerve endings. J Autonom Nerv System 68: 43-48, 1998
15. Yahagi N, Watanabe Y, Kumon K: Ultrasound detection of diaphragmatic paralysis after cardiac surgery.
Ann Thorac Surg 65: 1841, 1998
16. Yahagi N: Invited commentary for “A randomized,
double-blind trial of inhaled nitric oxide in LVAD
recipients with pulmonary hypertension”. Ann
Thorac Surg 65: 345, 1998
17. Haruna M, Kumon K, Yahagi N, Watanabe Y: Blood
volume measurement at bed side using ICG pulse
spectrophotometry. Anesthesiology 89: 1322-1328,
1998
18. Kumon K, Yahagi N. Nitric oxide inhalation
increased oxygen delivery after cardiovascular surgery in adult patients whether or not they have pulmonary hypertension. J.Anesth 12: 180-184, 1998
19. Yamazaki T, Akiyama T, Kawada T, Kitagawa H,
Takauchi Y, Yahagi N, Sunagawa K: Norepinephrine
efflux evoked by potassium chloride in cat sympathetic nerves: dual mechanism of action. Brain Res
794: 146-150, 1998
20. Nakayama M, Kumon K, Yahagi N, Haruna M,
Watanabe Y, Hayashi H: Antiphospholipid antibody
syndrome in a case with redo coronary artery bypass
grafting under cardiopulmonary bypass. Surg Today
28: 423-426, 1998
21. Yahagi N, Kumon K, Tanigami H, Watanabe Y,
Haruna M, Matsui J: Helium/Oxygen breathing
improved hypoxemia after cardiac surgery. Artif
Organs 21: 24-27, 1997
22. Yahagi N, Kumon K, Watanabe Y, Tanigami H,
Haruna M, Hayashi H, Imanaka H, Takeuchi M,
Murakami A: Detection of tracheal stenosis bycineangiocardiography in patients with congenital
heart disease. Anesth Analg 85: 1180-1181, 1997
23. Matsui J, Yahagi N, Kumon K, Hayashi H, Watanabe
Y, Haruna M, Yagihara T, Takamoto S: Effect of
inhaled nitric oxide on postoperative pulmonary circulation in patients with congenital heart disease.
Artif Organs 21: 17-20, 1997
24. Tanigami H, Yahagi N, Kumon K, Watanabe Y,
Haruna M, Matsui J, Hayashi H: Long-term sedation
with isoflurane for patients after cardiac surgery.
Artif Organs 21: 21-23, 1997
25. Hayashi H, Kumon K, Yahagi N, Haruna M,
Watanabe Y, Matsui J, Hattori R: Successful treatment of mediastinitis after cardiaovascular surgery
using electrolyzed strong acid aqueous solution. Artif
Organs 21: 39-42, 1997

296

26. Yamazaki T, Kawada T, Akiyama T, Kitagawa H,
Takauchi Y, Yahagi N, Sunagawa K: Omega-conotoxin GVIA and desipramine insensitive norepinephrine
efflux from cardiac sympathetic nerve terminal.
Brain Res 761: 329-332, 1997
27. Murakami A, Fukahara K, Ichida F, Yahagi N,
Kumon K: Significance of nitric oxide inhalation forreperfused lung. Artif Organs 21: 83-84, 1997
28. Watanabe Y, Kumon K, Yahagi N, Haruna M,
Hayashi H, Matsui J: Use of a pulse dye-densitometry
ICG-test (indocyanine green clearance test) to evaluate liver cirrhosis before cardiac surgery. Comparison
with Child’s classification. World Congress Intensive
Crit Care Med 9: 94, 1997
29. Watanabe Y, Kumon K, Yahagi N, Haruna M,
Hayashi H, Matsui J: Assessment of hepatic function
using a pulse dye-densitometry ICG-test (indocyanine green clearance test): value in patients withdeteriorated liver function after cardiac surgery. 9thEuropean Congress on Intensive Care Medicine pp797-801, 1996
30. Yahagi N, Kumon K, Tanigami H, Watanabe Y,
Matsui J: Helium/Oxygen breathing improved
hypoxemia after cardiac surgery: Case reports.
Anesth Analg 80: 1042-1045, 1995
31. Yahagi N, Kumon K, Nakatani H, Matsui J, Sasako Y,
Isobe F, Sakakibara Y, Kitoh Y, Nagata S, Haruna M,
Watanabe Y, Takamoto S: Inhaled nitric oxide forthe management of acute right ventricular failure inpatients with a left ventricular assist system. Artif
Organs 19: 557-558, 1995
32. Yahagi N, Tanigami H, Watanabe Y, Kumon K: A
problem with an infant speaking tracheotomy tube
(Vocalaid”). Anaesthesia 50: 91-92, 1995
33. Yahagi N, Kumon K, Umemoto T, Shimura H,
Kawaguchi AT, Tanigami H, Miyashita K: Cyclic
decrease in mixed venous oxygen saturation for the
early diagnosis of seizure complications after cardiac
surgery. Anesth Analg 80: 404-407, 1995
34. Yahagi N, Kumon K, Tanigami H, Watanabe Y, Sato
T: A new, simple device for fixing orotracheal tubes.
Anaesthesia 49: 916, 1994
35. Yahagi N, Kumon K, Tanigami H, Watanabe Y,
Ishizaka T, Yamamoto F, Nishigaki K, Matsuki O,
Yagihara T: Inhaled nitric oxide for the postoperative
management of Fontan-type operation. Ann Thorac
Surg 57: 1371-1372, 1994
36. Yahagi N, Furuya H, Matsui J, Sai Y, Amakata Y,
Kumon K: Improvement of the left Boncho-Cath”
double lumen tube. Anesthesiology 81: 781-782,
1994
37. Yahagi N, Kumon K, Nakatani T, Ishikawa T,
Tanigami H, Eishi K, Takahashi S: Peripartum cardiomyopathy and tachycardia followed by multiple
organ failure. Anesth Analg 79: 581-582, 1994
38. Yahagi N, Tanigami H, Kumon K, Yamada K,
Takahashi O, Yagihara T: Tracheal granulations secondary to wall compression by endotracheal tubes in
infants. Can J Anaesth 41: 1123-1124, 1994

39. Yahagi N, Kumon K, Tanigami H: Bronchial lavage
with a fibreoptic bronchoscope via a laryngeal mask
airway in an infant. Anaesthesia 49: 450, 1994
40. Murakami A, Fukahara K, Uozaki Y, Ichida F,
Tsubata S, Yagihara T, Yahagi N. Three cases of left
ventricular outflow tract obstruction after intracardiac repair of congenital heart disease. Advanced
Cardio-Thoracic Surg , Axel Springer Japan
Publishing. pp11-4, 1994
41. Yahagi N, Kumon K, Sugimoto H, Inagaki Y: A use oftelescope of rigid type bronchoscope through endotracheal tube in infants. Anesth Analg 76: 207, 1993
42. Kumon K, Sugimoto H, Kitahara H, Yahagi N:
Application of recombinant human erythropoietin
for preoperative autologous blood banking on
patients undergoing cardiac surgery. Therapeutic
Plasmapheresis (Xll), pp 875-878, 1993
43. Inagaki Y, Kumon K, Sugimoto H, Yahagi N: A new
hemofiltration device with reversible roller pump forcorrecting volume controlled by a precise weight balancer. Therapeutic Plasmapheresis (Xll), pp 757-759,
1993
44. Yahagi N, Nishikawa A, Matsui S, Komoda Y, Sai Y,
Amakata Y: Pituitary apoplexy following cholecystectomy. Anaesthesia 47: 234-236, 1992
45. Yahagi N, Nishikawa A, Sai Y, Matsui J, Amakata Y:
Double aortic arch presenting as massive haematemesis after removal of a nasogastric tube. Can J
Anaesth 39: 894, 1992
46. Yahagi N, Furuya H, Sai Y, Amakata Y: Effect of
halothane, fentanyl, and ketamine on the thresholdfor transpulmonary passage of venous air emboli in
dogs. Anesth Analg 75: 720-723, 1992
47. Stuhmar W, Conti F, Suzuki H, Wang X, Noda M,
Yahagi N, Kubo H, Numa S: Structural parts involvedin activation and inactivation of the sodium channel.
Nature 339: 597-603, 1989
48. Suzuki h, Bechk S, Kayano T, Noda M, Kubo H,
Yahagi N, Numa S: Functional expression of cloned
cDNA encoding sodium channel lll. FEBS Letters
228: 195-200, 1987
49. Yahagi N, Furuya H: The effects of halothane andpentobarbital on the threshold of transpulmonary
passage of venous air emboli in dogs. Anesthesiology
67: 905-909, 1987
50. Yahagi N, Miyazaki M, Itoh Y: Cardiopulmonary
effect of halothane concentration on pulmonary circulation during air embolism in dogs. Resuscitation
13: 81-86, 1986297

Department of
Mental Health
Outline and Research Objectives
Outline
The department was first established as the 4th Department of Clinical Medicine andNursing in the School of Health Care and Nursing in the Faculty of Medicine University ofTokyo in 1953. The department was renamed to the department of Mental Hygiene according to the reorganization of the School of Health Care and Nursing into the School of Health
Sciences in 1965. In 1992, the department underwent second renaming to the Departmentof Mental Health and Psychiatric Nursing by the reorganization of the School of Health
Sciences into the School of Health Sciences and Nursing. In 1997, according to the restructuring of the Faculty of Medicine into the Graduate School of Medicine, the Department of
Mental Health and Psychiatric Nursing was divided into the Department of Mental Healthand the Department of Psychiatric Nursing.

Research Objectives
The research objectives of this department are to conduct clinical psychosocial studies inthe vast field of mental health and to disseminate results of those studies internationally.
For the last five decades, the department has had many clinical research projects, forexample, psychotherapy, forensic psychiatry, community mental health, mental health/psychiatric services, psychiatric rehabilitation, child and adolescent mental health, and developmental disorders, depending on the specialty and interests of faculty members, who haveconducted those studies in collaboration with researchers in and outside Japan by employing psychosocial methodologies adopted from diverse allied disciplines (i.e., psychiatry, clinical psychology, nursing, education and sociology).
For the past five years, this department has conducted clinical studies with close collaboration with the Department of Psychiatric Nursing established in 1997. The Department of
Psychiatric Nursing has many clinical research projects of its own to substantiate clinicalactivities in psychiatric nursing: time and motion studies of nurses in acute psychiatricwards; clinical competency of psychiatric nursing; skills of community psychiatric nursing;mother’s mental health and child abuse; issues of spirituality with cancer patients; and careburden of families with ALS clients.
For the last decade, the Department of Mental Health has concentrated its research
activities mainly in 5 areas: (a) developmental disorders, (b) child and adolescent mental
health, (c) mental health services, (d) geriatric psychiatry/mental health, and (e) social psychiatry/community mental health, as introduced in the past research and major accomplishment section.

Faculties and Students

Past Research and Major Accomplishments

Professor and Chair Hiroshi Kurita, MD, PhD (since 1992)
Associate Professor in Mental HealthIwao Oshima, PhD (since 1996)
Associate Professor in Psychiatric Nursing
Mami Kayama, PhD (since 2002)
Associate .................................1
Graduate student ..................37
Research student.....................8
Secretary .................................4

The major accomplishments of this department
are summarized below in 5 areas, with the number ofa related article listed in the select publications section in parentheses.

298(a) Developmental disorders (1-16)
In this area, studies on Childhood disintegrative
disorder (CDD) and related conditions (1, 5, 9) are particularly of note. CDD is an autistic condition of
unknown etiology characterized by severe metalregression emerged after normal mental development
at least by age 2 followed by a severe autistic andmentally retarded state. Prof. Kurita, one of leading
clinical researchers of CDD, has conducted diagnostic/nosological studies on CDD or its synonym, disintegrative psychosis (DP), in comparison with autismthat could show similar yet much milder regression
than CDD/DP. Those studies contributed to the adoption of CDD as a type of pervasive developmental disorders (PDD) in the two international diagnostic systems: the Diagnostic and Statistical Manual of Mental
Disorders 4th edition (DSM-IV) (American Psychiatric
Association, 1994) and the International Classificationof Diseases and Related Health Problems 10the revision (ICD-10) (World Health Organization, 1993).
(b) Child and adolescent mental health (17-24)
We conducted studies aiming at clarifying theprevalence of behavior and emotional problems in
Chinese children and their risk factors by using a
Chinese version of the Child Behavior Checklist(CBCL) in collaboration with the Department of
Psychiatry, Shandong Medical School, China. The
studies participated by about 3,000 children and adolescents and their parents and teachers in Shandong
province yielded a series of outstanding papers. The
findings in those studies would contribute to the promotion of mental health of Chinese children and adolescents.
(c) Mental health services (25-34)
Studies in this area clarified present status andchallenges of mental health services: service systems(26, 27, 30, 33); evaluation of services (28, 32, 34); andmental health problems in professionals (29, 31).
(d) Geriatric psychiatry/mental health (35-42)
Studies in this area clarified early neuropsychological deficits of patients with dementia (37, 38); pathophysiology of Alzheimer’s disease (35, 40, 41); mental
health problems of the elderly (39); and mental healthof caregivers of patients with dementia (36, 42).
(e) Social psychiatry/community mental health(43-46)This area involves studies on alcohol dependency(43), mental health problems of sexual harassment
victims (44), and supporting patients with schizophrenia (45, 46).

Current Research(a) Screening pervasive developmental disorders
Pervasive developmental disorders (PDD) are agroup of autistic conditions adopted in the current
international classification systems of mental disorders, DSM-IV and ICD-10. The prevalence of PDD wasrecently reported as high as 0.6%.
The early detection of PDD is important, since
early intervention is suggested as effective in ameliorating autistic symptoms and facilitating developmentof children with PDD even if the cure is not possible.
For early detection of PDD, we developed a Japaneseversion of CHAT (Checklist for Autism in Toddlers), a
promising screening tool for PDD developed in UK,
and are now gathering clinical data. We also used theTokyo Autistic Behavior Scale (TABS) developed by us
and the Infant Behavior Checklist (IBC), both of whichrate autistic behavior in infancy, in the same clinicalfields as the CHAT data are gathered to collect data on
early features of PDD. Based on these data, we areplanning to develop an efficient screening system for
PDD.
(b) Early development of high-functioning PDD
High-functioning PDD (HPDD) is a PDD with an
IQ over 70 (i.e., not mentally retarded). Although such
PDD was considered rare until late 1980s, more than
50% of PDD patients are now suggested to be HPDD.
However, social and vocational outcomes of HPDD
patients are quite unsatisfactory due to a lack of adequate programs for their education and social adaptation. It is quite unknown how HPDD differs frommentally retarded PDD in early developmental characteristics. Since such features would help early identification of HPDD among PDD, we are now gathering
relevant data in young PDD children by using theTABS, IBC and other scales.
(c) Adult outcomes of PDD patients
PDD is a life-long condition but its adult outcomesare still unclear, since many of long-term outcome
studies of PDD were conducted before 1990, when
remedial education and social support systems were
not well established. Since we have one of the largest
data bases on PDD in the world and have followed-up
a large number (more than 150) of adult PDD patients(over age 20). We are now conducting a comparative
study of adult outcomes between autistic disorder(prototypical PDD) and other PDD. We are also planning a study to clarify prognostic factors of adult outcomes of PDD.
(d) Difference and similarity of high-functioning
PDD and attention deficit/hyperactivity disorder(ADHD)In clinical settings, it is frequently difficult to differentiate between high-functioning PDD (HPDD) and
ADHD, though difference in treatment does exist
between them. We are now conducting a comparative299

study between HPDD and ADHD to identify clinicalvariables to differentiate between both conditions.

Future Prospects
The most important thing for the department in
the foreseeable future is to facilitate research activities
on mental health more intensively and extensively incollaboration with researchers in and outside Japan.
Since the current professor will retire in 2 years fromnow and in view of the diversity and broadness of the
field of mental health, it is difficult to prospect fixedresearch themes in the future. However, it is crucially
important to have a world-class clinical researcher
who can perform excellent studies on mental healthin his/her own specialty and educate graduate students based on those research activities. Such a person would come from any specialty relating to mental
health.
It is also important to facilitate psychiatric nursing
studies in collaboration with the Department of
Psychiatric Nursing established in 1997, which has
published many quality Japanese papers based on its
clinical studies and is now concentrating to publish
English papers. Since many graduate students haveparticipated in research projects in both departments,
it is productive to continue the close collaboration of
the two departments in clinical studies.

Research Grants (for the past 5 years)
1. The Research Grant (8B-3) for Nervous and Mental
Disorders (the Ministry of Health and Welfare).
Title: The study on pathophysiology and treatment
of disorders of behavioral, emotional and psychological development from infancy to adolescence. Term:
1996-1998. Head investigator: Hiroshi Kurita, MD
2. Grant-in-Aid for Scientific Research (C) (2) (theMinistry of Education, Science, Sports and Culture).
Title: A clinical study on symptoms, development
and course of childhood disintegrative disorders.
Term: 1997 to 2000. Head investigator: Hiroshi
Kurita, MD
3. Grant-in-Aid (11A-6) for Research on Nervous andMental Disorders (the Ministry of Health, Labourand Welfare). Title: A study on the relationshipbetween attention-deficit/hyperactivity disorder andpervasive developmental disorders. Term: 19992001. Investigator: Hiroshi Kurita, MD
4. Grant-in-Aid for Scientific Research (C) (2) (theMinistry of Education, Culture, Sports, Science andTechnology). Title: A nosological and diagnostic
study on other pervasive developmental disorders.
Term: 2001 to 2003. Head investigator: Hiroshi
Kurita, MD
5. Grant-in-Aid (14A-8) for Research on Nervous andMental Disorders (the Ministry of Health, Labourand Welfare). Title: A study on the difference and300

similarity of attention-deficit/hyperactivity disorderand pervasive developmental disorders. Term: 20022004. Investigator: Hiroshi Kurita, MD

Select Publications(a) Developmental disorders
1. Kurita H. Infantile autism with speech loss before
the age of thirty months. Journal of the American
Academy of Child Psychiatry 24, 191-196, 1985.
2. Kurita H, Uchiyama T and Takesada M. Tokyo Child
Development Schedule. I. Test-retest reliability andconcurrent validity. Folia Psychiatrica et Neurologica
Japonica 39, 129-138, 1985.
3. Kurita H. Variables relating to the mental development of children with infantile autism. Japanese
Journal of Psychiatry and Neurology 40, 161-168,
1986.
4. Kurita H. A case of Heller’s syndrome with school
refusal. Journal of Autism and Developmental
Disorders 18, 315-319, 1988.
5. Kurita H. The concept and nosology of Heller’s syndrome: Review of articles and report of two cases.
Japanese Journal of Psychiatry and Neurology 42,785-793, 1988.
6. Kurita H, Miyake Y and Katsuno K. Reliability andvalidity of the Childhood Autism Rating Scale—Tokyo Version (CARS-TV). Journal of Autism andDevelopmental Disorders 19, 389-396, 1989.
7. Kurita H and Miyake Y. The reliability and validity of
the Tokyo Autistic Behavior Scale. Japanese Journalof Psychiatry and Neurology 44, 25-32, 1990.
8. Kurita H. School refusal in pervasive developmental
disorders. Journal of Autism and Developmental
Disorders 21, 1-15, 1991.
9. Kurita H, Kita M and Miyake Y. A comparative studyof development and symptoms among disintegrative
psychosis and infantile autism with and without
speech loss. Journal of Autism and Developmental
Disorders 22, 175-188, 1992.
10. Kurita H, Shiiya J and Ito H. A nationwide survey of
day-care for children with mental retardation in
Japan. Japanese Journal of Psychiatry and Neurology
48, 57-63, 1994.
11. Kurita H and Nakayasu N. An autistic male presenting seasonal affective disorder (SAD) and trichotillomania. Journal of Autism and Developmental
Disorders 24, 687-692, 1994.
12. Kurita H, Saito T and Kita M. Regression in mental
development following a psychosocial stressor in disintegrative psychosis. Shimizu M (Ed.). Recent
Progress in Child and Adolescent Psychiatry, pp.2128, Springer, 1996.
13. Kurita H. Specificity and developmental consequences of speech loss in children with pervasive
developmental disorders. Psychiatry and Clinical
Neurosciences 50, 181-184, 1996.

14. Kurita H. A comparative study of Asperger syndrome
with high-functioning atypical autism. Psychiatry
and Clinical Neurosciences 51, 67-70, 1997.
15. Kurita H. Delusional disorder in a male adolescentwith high-functioning PDDNOS. Journal of Autism
and Developmental Disorders, 29:419-423, 1999.
16. Tachimori H, Osada H, Kurita H. Childhood Autism
Rating Scale--Tokyo version (CARS-TV) for screening
pervasive developmental disorders (PDD). Psychiatry
and Clinical Neurosciences (in press).

(b) Child and adolescent mental health
17. Liu XC, Ma DD, Kurita H and Tang MQ. Self-reported depressive symptoms among Chinese adolescents.
Social Psychiatry and Psychiatric Epidemiology 34,
44-47, 1999.
18. Mori M. The influence of father-daughter relationship and girl’s sex-roles on girl’s self-esteem.
Archives of Women’s Mental Health 2, 45-47, 1999.
19. Liu XC, Kurita H, Guo CQ, Miyake Y, Ze J and Cao
HL. Prevalence and risk factors of behavioral andemotional problems among Chinese children aged 611. Journal of the American Academy of Child andAdolescent Psychiatry 38, 708-715, 1999.
20. Liu XC, Kurita H, Sun ZX and Wang FX. Risk factorsof psychopathology among Chinese children.
Psychiatry and Clinical Neurosciences 53, 497-503,
1999.
21. Liu XC, Uchiyama M, Okawa M and Kurita H.
Prevalence and correlates of self-reported sleep problems among Chinese adolescents. Sleep 23, 27-34,
2000.
22. Liu XC, Kurita H, Guo CQ, Tachimori H, Ze J andOkawa M. Behavioral and emotional problems in
Chinese children: Teacher reports for ages 6 to 11.
Journal of Child Psychology and Psychiatry 41, 253260, 2000.
23. Liu XC, Guo CQ, Okawa M, Zhai J, Li Y, Uchiyama
M, Neiderhiser JM and Kurita H. Behavioral andemotional problems in Chinese children of divorced
parents. Journal of the American Academy of Childand Adolescent Psychiatry 39, 896-903, 2000.
24. Liu XC, Kurita H, Uchiyama M, Okawa M, Liu LQ
and Ma DD. Life events, locus of control, and behavioral problems among Chinese adolescents. Journalof Clinical Psychology 56, 1565-1577, 2000.
(c) Mental health services
25. Kumakura N, Ito H, Mori T, Saito T, Kurisu E, Asaka
A, Sasaki Y, Rhi B, Kim Y, Ha K, Rin H and Yang LY.
Attitudes change towards mental illness during nursing education: A cross-cultural study of student nurses in Korea, Republic of China and Japan. Asia-Pacific
Journal of Public Health 6, 120-125, 1992/1993.
26. Ito H, Mori T, Kumagai N, Miyauchi M, Harada S,
Yamamura M, Nakano R, Kurita H and Sasaki Y.
Changing pattern of mental health utilization in the
Hachijojima. Psychiatry and Clinical Neurosciences49, 185-187, 1995.

27. Ito H, Kishi Y and Kurosawa H. A preliminary studyof staff perception of psychiatric services in general
hospitals. General Hospital Psychiatry 21, 57-61,
1999.
28. Ito H, Shingai N, Yamazumi S, Sawa Y and Iwasaki S.
Characteristics of nonresponders to a patient satisfaction survey at discharge from psychiatric hospitals. Psychiatric Services 50, 410-412, 1999.
29. Ito H, Kurita H and Shiiya J. Burnout among directcare staff members of facilities for persons with mental retardation in Japan. Mental Retardation 37, 477481, 1999.
30. Ito H, Eisen SV, Sederer LI and Tachimori H.
Involuntary admission in Japanese psychiatric hospitals. International Medical Journal 7, 109-112, 2000.
31. Ito H, Eisen SV, Sederer LI, Yamada O andTachimori H. Factors affecting psychiatric nurses’
intention to leave their current job. PsychiatricServices 52, 232-234, 2001.
32. Ito H, Tachimori H, Miyamoto Y and Morimura Y.
Are the care levels of people with dementia correctly
assessed for eligibility of the Japanese long-term careinsurance? International Journal of Geriatric
Psychiatry 16, 1078-1084, 2001.
33. Ito H, Eisen SV and Sederer LI. Acute care psychiatryat McLean Hospital, Massachusetts. International
Medical Journal 8, 91-96, 2001.
34. Ito H and Sederer LI. Are publicly-insured psychiatricoutpatients in Japan satisfied? Health Policy 56, 205213, 2001.

(e) Geriatric psychiatry/mental health
35. Dai XY, Nanko S, Hattori M, Fukuda R, Nagata K,
Isse K, Ueki A and Kazamatsuri H. Association of
apolipoprotein E4 with sporadic Alzheimer’s diseaseis more pronounced in early onset type.
Neuroscience Letters 175, 74-76, 1994.
36. Matsuda O. The effect of coping on the caregiver of
elderly patients with dementia. Psychiatry andClinical Neurosciences 49, 209-212, 1995.
37. Matsuda O, Saito M and Sugishita M. Cognitive
deficits of mild dementia: A comparison between
dementia of the Alzheimer’s type and vasculardementia. Psychiatry and Clinical Neurosciences 52,
87-91, 1998.
38. Matsuda O and Saito M. Features of Alzheimer’s disease: Crystallized and fluid intelligence in elderly
patients with mild dementia of the Alzheimer type.
International Psychogeriatrics 10, 147-154, 1998.
39. Hashimoto K, Kurita H, Hratani T, Fujii K and
Ishibashi T. Direct and buffering effects of social
support on depressive symptoms of the elderly withhome help. Psychiatry and Clinical Neurosciences 53,
95-100, 1999.
40. Asada T, Motonaga T, Yamagata Z, Uno M and
Takahashi K. Association between retrospectively
recalled napping behavior and later development of
Alzheimer’s disease: Association with APOE genotypes. Sleep 23, 629-634, 2000.

301

41. Asada T, Motonaga T and Kinoshita T. Predictors ofseverity of behavioral disturbance among community-dwelling elderly individuals with Alzheimer’s disease: A 6-year follow-up study. Psychiatry andClinical Neurosciences 54, 673-677, 2000.
42. Miyamoto Y, Ito H, Otsuka T, Kurita H. Caregiverburden in mobile and non-mobile demented
patients: A comparative study. International Journalof Geriatric Psychiatry 17, 765-773, 2002.

(d) Social psychiatry/community mental health43. Matsuoka K, Kim Y, Toshida S and Oshima N.
Relationship between age of onset, antisocial history
and general psychopathological traits in Japanese
alcoholics. Psychiatry and Clinical Neurosciences 54,
413-417, 2000.
44. Matsuoka K, Kurita H, Abe R and Osada H. Mental
health of sexual harassment victims in Japanesenurses. Archives of Women’s Mental Health 4, 1-4,
2001.
45. Kayama M, Zerwech J, Thornton K and Murashima
S. Japanese expert public health nurses empower
clients with schizophrenia living in the community.
Journal of Psychosocial Nursing and Mental HealthServices 39, 40-45, 2001.
46. Oshima I, Ito J, Taira N. Need for an integrated program of case management and pychoeducation inlong stay psychiatric patients in Japan. Kashima H,
Falloon IRH, Mizuno M and Asai M. (eds).
Comprehensive Treatment of Schizophrenia: Linking
neurobehavioral findings to psychosocial approaches.
Springer, Tokyo, pp.196-203, 2002.

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Department Of
Social Gerontology
Outline And Research Objectives
It is often voiced from the general public that recent advancement of medicinal technology would not necessarily lead to the happiness of people: Life prolongation technology
enables even the terminally ill to live for a considerable period. How to use the technology
is a serious problem in clinical practice. Also, there is evidence that the prolongation of life
expectancy for the elderly does not mean the prolongation of health and productivity, butthat of morbidity.
Taking another example, we are experiencing ethical dilemmas with the application of
medical technology such as genetic screenings and organ transplantation. When we turn to
the worldwide situation regarding health, we will find poverty and unequal distribution interms of health resources and outcomes.
The department is studying these health-related problems from psycho-social-ethicalperspectives, many of which are often difficult to decide upon. Major topics include elderly
health, cancer and/or terminal care, international health and bio-medical ethics among others. We are currently conducting several research projects as described below.

Faculties and Students
Professor and Chair

Ichiro Kai, M.D., M.P.H. (since
1997)
Associate Professor and Lecturer None
Associate .................................2
Visiting Researcher ...............22
Graduate student ..................18
Research student.....................3

Past Research And Major Accomplishments
1. Elderly HealthThis is the major field of interest of our department. Several most important researches will bebriefly described.
1) Psychotic manifestations of the demented elderly and their relationship with caretakers
We examined the correlation between the psychotic symptoms of the demented elderly and theirrelationship with caretakers. We developed a scalemeasuring the relationship (CPR Scale), and conductedsurvey in three culturally different areas, Tokyo,
Nagano and Okinawa. We found the better relationship indicated by higher CPR score led to less psychotic symptoms. The results suggest that the psychotic
manifestations of dementia can be influenced by environmental factors. (References 2-5, 12,17)2) Functional capacity and active life expectancy(ALE) of the elderly
The health status of a population is traditionallymeasured by mortality. However, indices based on
mortality are increasingly unsatisfactory to assess
health status, especially in an aging society. We conducted survey on functional capacity of the elderly
and its chronological changes, using a large-scalecohort of the community residents in Nagano
Prefecture. We calculated active life expectancy (ALE)and suggested it could be a useful health indicator for
elderly population. (References 16,18, 37-38, 46)3) Reciprocity of social support on subjective
well-being of the elderly
Traditional support study emphasizes the importance of receiving support. We examined the patternof support exchange (i.e., receiving and providing) andits effects on the subjective well-being of the elderly inrural Japan as well as a number of Asian countries
such as Korea, Malaysia and Indonesia. In Korea, both
receiving and providing support correlated to better
subjective well-being, while only providing did inJapan. We concluded the meaning of receiving support
were different between the two cultures. The findings
led to further intervention study, as described later inthis document. (References 23, 41-42)
4) Long-term care of the elderly
Long-term care of the elderly becomes increasingly
important medical/welfare issue in Japan. Geriatricintermediate care facilities (GICFs) were established in1986 to facilitate discharge from hospitals and sup-303

port home care. Since then, we have conducted surveys on how the GICFs were used in Japan, and proposed the policy for their effective utilization.
(References 20, 22, 39)
2. Cancer Care and Terminal Care
We conducted several surveys on cancer/terminal
care with special emphasis on psycho-social and quality of life (QOL) aspects. We emphasize the importanceof effective communication between patients andhealth care providers. Recently, we are interested insocio-cultural aspects of sexuality after cancer therapy.
The latter is partly supported by Grant-in-Aid for
Scientific Research from Japanese Society for
Promotion of Science (JSPS). (References 14, 19, 24,
34, 36, 40)
3. International Health
The majority of the studies regarding international
health were done while I was affiliated with
Department of International Community Health,
School of International Health, from 1992 to 1997.
We were particularly interested the prevention of
AIDS in Japan and the other countries in Asia.
(References 10-11, 13, 15, 25, 28-29)
4. Bio-medical Ethics
The majority of the studies regarding bio-medical
ethics were done in collaboration with Prof. Akira
Akabayashi, who was a lecturer of our department
and is now the chair of Department of Health
Economics. Most studies are questionnaire surveys,
and the topics include the ethics education in medical
schools and the role of ethics committees among others. (References 21, 26-27, 31, 33, 35, 43-44)

Current ResearchTwo of the ongoing research projects of the department will be described below.
1. Intervention Study on Social Support Exchange
Our previous study suggests that providing support to spouse, children and friends is important topromote the subjective well-being of the elderly. We
devised an intergenerational program involving theelderly and college students. The intervention was
basically an interview in which the elderly were supposed to help the college students by telling their life
history to the students and, in turn, were benefitedby the interview. We found the majority of outcome
indices such as life satisfaction and the attitudes
toward young generation had improved considerably
after the intervention. This study was partly supported by Grant-in-Aid for Scientific Research from JSPS.

304

2. Frequency of Going Outside and Risk ofFunctional Decline among the Frail Elderly
To be house-bound is hypothesized to heighten
the risk of functional and intellectual deterioration
among the frail elderly. We followed up a cohort of
the frail elderly in Nagano Prefecture, and examined
the relationship between the frequency of going outside home and various physical and psychological outcomes indices. Even after controlling a number of confounding factors, going outside home was shown toinversely correlate to deterioration in physical andpsychological health one-year later. These findings
suggest the importance of preventing house-boundstatus of the frail elderly. Subsequently, we are conducting an intervention study involving home visitingby public health nurses. This study is partly supported by Grant-in-Aid for Longevity Study from Ministryof Health, Labor and Welfare and Mitsubishi
Foundation.

Future Prospects
Two of the possibly important research fields forour department will be briefly mentioned below.
1. Study on Successful Aging and Productivity
Successful aging is an important concept in thefield of contemporary gerontology. One of the key factors to enable successful aging is productivity. We
plan to conduct survey on productivity includingsalaried work, household work and volunteer, and its
relationship with the health of the elderly. Large-scaleintervention study on prosocial behaviors by the elderly (e.g., providing support to others) will be done aswell.
2. Effective Policy for Long-term Care
Since the establishment of Long-term Care
Insurance (LCI) in 2000, care of the frail elderly has
become important social issue. In future, we plan toput more emphasis on practical aspects and policy
issues in social gerontology. We plan to conduct alarge-scale survey on LCI users in Tokyo Metropolitan
Area and make recommendations on the policy for
effective long-term care in Japan.

Research Grants
1. Intervention study on social support exchange: Anintergenerational program involving the elderly andhigh school students. Grant-in-Aid for Scientific
Research (KAKENHI) from JSPS (three years from1998 to 2001) 2.8 million yen
2. Change and its related factors in sexuality forpatients experiencing breast cancer surgery. Grant-

in-Aid for Scientific Research (KAKENHI) from JSPS
(three year from 2000 to 2003) 3 million yen
3. Frequency of going outside home and functional
decline among the frail elderly. Grant-in-Aid forLongevity Study (KAKENHI) from Ministry of
Health, Labor and Welfare (three years from 2000 to2003) 5.35 million yen
4. Effectiveness of a home visiting program for thehouse-bound elderly. Mitsubishi Foundation (twoyears from 2001 to 2003) 2 million yen
5. Predictors for frailty and/or mortality among therural elderly. Daiwa Health Foundation (one year
from 2001 to 2002) 1 million yen

Select Publications(1) Original Articles
1. Kai I, Inaba Y: Statistical analysis on large-scale foodpoisoning outbreaks of unknown etiology. Food
Hygiene Study 29(12):1031-1040, 1979 (in Japanese)
2. Ohi G, Miyama T, Kai I, et al.: Psychotic manifestations in the bedfast elderly: The role of intellectual
impairment and environmental stress. Jap. J.
Gerontology 6(2):124-138, 1984 (in Japanese)
3. Kai I, Ohi G, Suzuki S: Applicability of Hasegawa’s
Dementia Rating Scale in rural areas in Japan:
Comparative study in Tokyo and Okinawa.
Gerontological Psychiatry 2(2):293-298, 1985 (inJapanese)
4. Ichikawa S, Nakagawa M, Ohi G, et al.: Developmentof caretaker-patient relationship assessment scale forthe frail elderly. Jap. J. Public Health 32(5):253-257,
1985 (in Japanese)
5. Miyama T, Suzuki S, Ohi G, et al.: Psychotic manifestations in the elderly with intellectual impairment:
Influence of the degree of their intellectual deterioration and caretaker-patient relationship. Jap. J. Public
Health 32(7):325-332, 1985 (in Japanese)
6. Nagahashi M, Yamazaki N, Ohi G, et al.: Dietary fiber
intake and diverticular disease of the colon: A casecontrol study. Jap. J. Hygiene 40(4):781-788, 1985 (inJapanese)
7. Takenaga N, Kai I, Ohi G: Evaluation of three cervicalcancer detection programs in Japan with special reference to cost-benefit analysis. Cancer 55(10):25142519, 1985
8. Yamaoka K, Ohi G, Miyama T, et al.: Burden of caretakers of the bedridden elderly: Development of aunidimensional scale. Jap. J. Public Health 33(6):279284, 1986 (in Japanese)
9. Ohi G, Hasegawa T, Kumano H, et al.: Why are cadaveric renal transplants so hard to find in Japan?: Ananalysis of economic and attitudinal aspects. Health
Policy 6:269-278, 1986
10. Ohi G, Kai I, Kobayashi Y, et al.: Epidemic potentialof AIDS and health policy in Japan. Jap. J. Public
Health 34(2):49-54, 1987 (in Japanese)
11. Kai I, Ohi G: Infectious disease specialists in academeand AIDS epidemics: Comparison of specialists inpublic health and clinical medicine. Jap. J. Public
Health 34(4):195-198, 1987 (in Japanese)
12. Takenaga N, Kai I, Ohi G, et al.: The caretakerpatient relationship and psychotic manifestations in
the bedridden elderly. Jap. J. Hygiene 42(2):563-569,
1987 (in Japanese)
13. Ohi G, Kai I, Kobayashi Y, et al.: AIDS prevention in
Japan and its cost-benefit aspects. Health Policy 8:1727, 1987
14. Kobayashi Y, Yamaoka K, Ohi G, et al.: The economic
analysis of home and hospital care for terminally illpatients. Jap. J. Public Health 35(1):11-18, 1988 (inJapanese)
15. Ohi G, Terao H, Hasegawa T, et al.: Notification of
HIV carriers: Possible effect on uptake of AIDS testing. Lancet II:947-949, 1988
16. Kobayashi Y, Kai I, Ohi G, et al.: A study on instrumental activities of daily living and its related factors
in the elderly in the farming community. Jap. J.
Public Health 36(4):243-249, 1989 (in Japanese)
17. Ohi G, Kai I, Ichikawa S, et al.: Psychotic manifestations in the bed-fast elderly: A preliminary communication. J. Human Ergology 18:237-240, 1989
18. Kai I, Ohi G, Kobayashi Y, et al.: Quality of life: A
possible health index for the elderly. Asia-Pacific J.
Public Health 5(3):221-227, 1991
19. Kai I, Ohi G, Yano E, et al.: Communication between
patients and physicians about terminal care: A survey in Japan. Soc. Sci. & Med. 36(9):1151-1159, 1993
20. Ishizaki T, Kai I, Hirayama T: Factors influencing
users’ destination upon discharge from a geriatricintermediate care facility in an urban area. Jap. J.
Geriatrics 32(2):105-110, 1995 (in Japanese)
21. Nishimura T, Miyasaka M, Akabayashi A, et al.: The
development of evaluation method for bioethics education in Japan: A questionnaire for measuring the
ethical problem identification ability. Bioethics
5(1):73-77, 1995 (in Japanese)
22. Ishizaki T, Kai I, Hisata M, et al.: Factors influencing
users’ return home on discharge from a geriatricintermediate care facility in Japan. J. Am. GeriatricSoc. 43(6):623-626, 1995
23. Kim HK, Lee SK, Hisata M, et al.: Social support
exchange and quality of life among the Korean elderly in a rural area. Jap. J. Public Health 43(1):37-49,
1996 (in Japanese)
24. Hisata M, Okazaki N, Kai I, et al.: Attitudes of outpatients toward informed consent in cancer treatment.
J. Jap. Soc. Cancer Therapy 31(3):171-185, 1996 (inJapanese)
25. Ohi G, Uemura I, Akabayashi A, et al.: Preventive
behaviors against HIV transmission adopted by
Japanese commercial sex workers (CSWs). Environ.
Health & Prev. Med. 2(3):132-134, 1997
26. Akabayashi A, Kai I, Itoh K, et al.: The acceptabilityof advance directive in Japanese society: A questionnaire study for healthy people in the physical checkup settings. Bioethics 7(1):31-40, 1997 (in Japanese)
27. Fukatsu N, Akabayashi A, Kai I: The current statusof ethics committees and decision making proce-305

dures in general hospitals in Japan. Bioethics
7(1):130-136, 1997 (in Japanese)
28. Nishimura-Takahashi T, Akabayashi A, Kai I, et al.:Social and behavioral factors associated with condom
use among female commercial sex workers (CSWs)
in Tarlac, the Philippines. Environ. Health & Prev.
Med. 2(4):167-171, 1998
29. Takai A, Wongkhomthong S, Akabayashi A, et al.:Correlation between history of contact with people
living with HIV/AIDS (PWAs) and tolerant attitudes
toward HIV/AIDS and PWAs in rural Thailand.
International J. STD & AIDS 9(8):482-484, 1998
30. Honda T, Kai I, Ohi G: Fat and dietary fiber intake
and colon cancer mortality: A chronological comparison between Japan and the US. Nutr. & Cancer
33(1):95-99, 1999
31. Akabayashi A, Miyasaka M, Kai I, et al.: Survey of
medical ethics education in Japanese medical
schools. Med. Education 30(1):47-53, 1999 (inJapanese)
32. Yokokawa Y, Kai I, Nakajima T: Development of a
“Self Efficacy for Health Promotion Scale” in community-dwelling elderly. Jap. J. Public Health 46(2):103112, 1999 (in Japanese)
33. Akabayashi A, Kai I: Study of 5th-year medical students interest in medical ethics and related factors.
Med. Education 30(2):77-82, 1999 (in Japanese)
34. Akabayashi A, Kai I, Takemura H, et al.: Truth
telling in the case of a pessimistic diagnosis in Japan.
Lancet 354(9186):1263, 1999
35. Miyasaka M, Akabayashi A, Kai I, et al.: Teaching
medical ethics: An international survey of medical
ethics curricula in Asia. J. Med. Ethics 25(6):514-521,
1999
36. Takahashi M, Kai I, Akabayashi A, et al.: Whoshould meet the needs of breast cancer patients?: Aquestionnaire survey of Japanese surgeons. Jap. J.
Breast Cancer 14(4):495-502, 1999 (in Japanese)
37. Ishizaki T, Kai I, Kobayashi Y: Estimation of active
life expectancy by Katz method: The results from
Saku Cohort Study. Health Indices 46(4):23-27, 1999
(in Japanese)
38. Ishizaki T, Kobayashi Y, Kai I: Functional transition
in instrumental activities of daily living among older
Japanese. J. Epidemiol. 10(4):249-254, 2000
39. Ishizaki T, Kai I: Risk of acute hospitalization among
long-term care patients. Jap. J. Geriatrics 37(6):472478, 2000 (in Japanese)
40. Watanabe Y, Akabayashi A, Ikeda T, et al.: An analysis of CPR decision-making by middle-aged and elderly healthy people in Japan. Bioethics 10(1):111-119,
2000 (in Japanese)
41. Kim HK, Kai I, Hisata M, et al.: Social support
exchange and subjective well-being among theJapanese elderly in a rural area. Jap. J. Gerontology
22(3):395-403, 2000 (in Japanese)
42. Kim HK, Hisata M, Kai I, et al.: Social support
exchange and quality of life among the Korean elderly. J. Cross-Cultural Gerontology 15(4):331-347, 2000

306

43. Watanabe Y, Kai I, Akabayashi A: Attitude of generalpublic toward genetic testing for colon cancer.
Bioethics 12:32-40, 2001 (in Japanese)
44. Akizuki Y, Kai I, Akabayashi A: Attitudes of the general public toward artificial insemination by donor.
Bioethics 12:62-70, 2001 (in Japanese)
45. Ohta M, Kai I: The development of a filial responsibility scale. Jap. J. Social Welfare 42(2):130-7, 2002
(in Japanese)
46. Ishizaki T. Kai I, Kobayashi Y, et al.: Functional transitions and active life expectancy for older Japaneseliving in a community. Arch. Gerontology &Geriatrics 35(2):107-119, 2002

(2) Other Papers (Books And Proceedings In English)
47. Ohi G, Kai I, Terao H, et al.: Cost-benefit aspects of
AIDS prevention programs: Its limitation in policy
making. in Fleming AF, et al. (eds.): The Global
Impact of AIDS , Alan R. Liss, Inc. (New York),
pp.251-262, 1988
48. Ohi G, Hisata M, Kai I: High-rise living and the elderly. in Takano T (ed.): Urban Health, Kyoiku Syoseki(Tokyo), pp.7-13, 1993
49. Ishizaki T, Kai I, Hisata M, et al.: Factors influencing
users’ destination upon discharge from a geriatricintermediate care facility. in Beregi E, et al. (eds.):
Recent Advances in Aging Science, Monduzzi Editore(Bologna), pp.2193-2196, 1993
50. Kai I.: Active life expectancy and its correlates among
the elderly living in a rural community in Japan. in
Hisamichi S, et al. (eds.): WHO-CC Monograph No.3
Active (Disability-Free) Life Expectancy , WHOCollaborating Center for Well-being in Aging(Sendai), pp.52-53, 1995

Department of
Health Economics /Health Promotion Sciences /Biomedical Ethics
Outline and Research Objectives
The former Department of Health Administration was established in 1967 and Dr.
Tsuneo Tanaka became its first professor in 1974. He devoted himself to the developmentof the community health care system in Japan and publishes numerous papers concerning
the social theory of health administration and the data management systems for community health care. He also contributed to the establishment of the School of Health Sciences. In
1985, Dr. Atsuaki Gunji became the second professor of the department. During Dr. Gunji’stenure, two major research projects were undertaken. One was “the effects of physical activity and inactivity on health.” From 1990, a 20- day bed rest human experimental study was
conducted in the context of an international cooperative research project that was supported by government grants. The other project concerned health care systems, especiallyhealth care economics and the quality of hospital care.
In 1996, the Department of Health Administration was devided into two departments:
the Department of Health Economics and the Department of Health Promotion Sciences.
Both were established as departments of the Graduate School of Medicine. In 1998, Dr.
Yasuki Kobayashi became professor of the Department of Health Economics. He conducted
research into health care delivery systems in Japan. In 2001, he moved to the Department of
Public Health. Since 1996, Dr. Kiyoshi Kawakubo has been taking charge of the Department
of Health Promotion Sciences.
In June 2002, Dr. Akira Akabayashi (now concurrently at Kyoto University) was appointed as Dr. Kobayashi’s successor. Dr. Akabayashi’s research area is in biomedical ethics and a
new Department of Biomedical Ethics will be officially established in April 2003.

Faculties and Students
Professor and Chair

Akira Akabayashi, M.D., Dr. Med.
Sc., (2002~, concurrently at Kyoto
University)
Associate Professors Kiyoshi Kawakubo, M.D., Dr.
Med. Sc., (1996~)
Associates ...............................1
Postdoctoral Fellows ........none
Graduate Students ..................7
Research Students...................2
Secretaries ...............................3

Past Research and Major Accomplishments
Past research can be divided into two fields, healtheconomics and health promotion sciences. In the fieldof health economics, analysis of terminal care cost for
the aged in a community, analysis of regional differences in medical care costs for the aged and analysis
of hospital care costs and revenues were the main
research topics. About 70% of the regional differences

was accounted for by the overall quantity of medicalcare supplies such as the number of hospital beds.
Other topics were related to the economic aspects ofhealth care and the quality of care. Both supply anddemand analyses were performed on, for example,
hospital costs and patient behavior. Tools for evaluating the quality of hospital care were also developed.
Recent research topics include studies related to either
or both the supply and demand sides of health care in
Japan such as physician distribution, outcomes of theseparation of pharmaceutical dispensing from medical
practice, and patient behavior under Japan’s national
health insurance system. The main research questions
are related to how the health and social systems
shape the health care systems and health itself, andwhat system should be established in order to maintain both efficiency and equity.
The focus of research activities of the Department
of Health Promotion Sciences is on the scientific basisof health promotion, specifically the effects of physical activity and inactivity on health status. Since 1991,
a bed rest study has been performed, involving the307investigation of the effects of simulated microgravity
on cardiovascular and metabolic parameters as well as
on bone density. This is the first bed rest study conducted in Japan. The “Healthy Japan 21“ strategy was
introduced in 2000 by the former Ministry of Healthand Welfare, and the department has devoted part ofits “physical activity and exercise” studies to this strategy. Currently the department is engaged in the development of health promotion strategies for the community and workplace. The assessment of health promotion resources in the community and workplace isthe major concern of these research activities.

Current ResearchWith the establishment of the new department of
Biomedical Ethics in April 2003, the foci of theresearch projects may shift to healthcare ethics andhealth promotion sciences (behavioral science).
Research in the field of health economics will also be
performed in close relationship with the Departmentof Public Health (chaired by Dr. Y. Kobayashi). The
research projects being conducted mainly by Dr.
Akabayashi are described below.
Grant-supported research projects (within 5
years)
1. Study of the methods for the formation of social
consensus related to advanced medical technology. The purposes of this project are: 1) to clarify
ethical, legal, and social issues related to theimplementation of new medical technology, and2) to explore methods for the formation of social
consensus and policy. This project includes thetheoretical aspects of social consensus, historicalcase analysis of organ transplantation and genetherapy in Japan, and three nationwide surveys.
Supported by the Special Coordination Fund for
Promoting Science and Technology, The Ministryof Education, Culture, Sports, Science andTechnology.
2. Publication of a medical ethics case book forJapan. The purpose of this project is to collectillustrative cases of ethical issues and to produce acasebook for the field of medical ethics in Japan,
which is hoped to become a standard textbook for
those who study or teach medical ethics as well asa resource for research. The book is also expected
to help explain current Japanese medical ethics tointerested persons in other countries. Supported
by a Grant-in-Aid for Scientific Research (B), The
Ministry of Education, Science, Sports andCulture.
3. Study of the functions and responsibilities of
ethics committees in Japan. The growing numberof ethics committees (ECs) indicates that signifi-308

cant changes have occurred in the decision making process in the clinical setting in Japan. The
main purposes of this study are:1) to investigate
the current status; 2) to make guidelines for establishing and running an EC in a general hospitalsetting; 3) to explore the extent of discretion andresponsibility of ECs, and 4) to make a comprehensive proposal.. Supported by a Grant-in-Aid for
Scientific Research (C), The Ministry of Education,
Science, Sports and Culture.
4. Comparative study of clinical ethics in the Asian
region. The purpose of this project is to engenderongoing inter and intra regional dialogue,
research, education and development of clinicalethics throughout the Asian region. Specifically,
this project will focus on issues that stem from thenature of the clinician-patient relationship, such as
how information is shared (informed consent,delivering bad news, role of the family, etc.), competence or decision-making capacity, confidentiality, and end-of-life decision making. The results
obtained through this preliminary study will be
used as the basis for a comprehensive study..
Supported by a Grant-in-Aid for Scientific
Research (C), The Ministry of Education, Science,Sports and Culture.
5. Others: Other research activities include research
into the acceptability of advance directives inJapan (a grant from the Ministry of Education in
1996-1997), the development of evaluation methods for biomedical ethics education (a grant from
the Ministry of Education in 1995-1996), and theethical and psycho-social aspects of living related
liver transplantation.

Future Prospects
Modern biomedical ethics in Japan has emerged asa new medical field and many medical schools are
establishing relevant departments or units. As long as
this discipline remains in the medical arena, it needs
to deal with both theoretical and practical issues.
Even though this discipline has not been fully established yet, many issues have already arisen. It is clearthat the focus will have to be on the development ofbasic teaching and research methods that can garnerthe support of both the medical community and society in general. Ethics committees constitute aninescapable part of the activities related to this discipline. Without sound ethics committees, no research
involving human subjects can be implemented.
Within this departments unit, health economics andbehavioral science make a major contribution to thestudy of biomedical ethics because allocation of healthcare resource is a major topic in healthcare ethics, andbehavioral science is essential for the teaching of com-munication skills with patients and an understandingof patient behavior. In the near future, this departments unit is hoped to become a domestic as well as
an Asian regional center for academic research.

Research Grants (within 5 years)
1. Study of the methods for the formation of social consensus related to advanced medical technology .
Special Coordination Fund for Promoting Scienceand Technology, The Ministry of Education, Culture,
Sports, Science and Technology 2001-2002,¥55,000,000 (PI: Akabayashi, A)
2. Study on the health promotion environmental
indices for the community: The Ministry of Health,
Labor and Welfare, Health Sciences Research Grant,
2000-2001, ¥55,000,000 (PI: Kawakubo, K)
3. Publication of a medical ethics case book for Japan.
Grant-in-Aid for Scientific Research (B), 1998-2000,
The Ministry of Education, Science, Sports andCulture, ¥3,600,000 (PI: Akabayashi, A)
4. Comparative study of clinical ethics in the Asian
region. Grant-in-Aid for Scientific Research (C), 2000,
The Ministry of Education, Science, Sports andCulture, ¥1,200,000 (PI: Akabayashi, A)
5. Study of the functions and responsibilities of ethics
committees in Japan . Grant-in-Aid for Scientific
Research (C), 1997-1999, The Ministry of Education,
Science, Sports and Culture, ¥2,200,000 (PI:Akabayashi, A)

Select Publications
1. Akabayashi A. Euthanasia, assisted suicide, and cessation of life support: Japan’s policy, law, and ananalysis of whistle blowing in two recent mercy
killing cases. Social Science & Medicine 55: 517-527,
2002.
2. Akabayashi A, Nishimori M, Fujita M, Slingsby BT.
Living related liver transplantation: a Japanese experience and development of a checklist for donor’s
informed consent. Gut 51: 2002 (in press)
3. Akabayashi A, Slingsby BT, Voltz R. Advance
Directives. In Bruera E. eds., Palliative Care in
Neurology Oxford University Press, New York, NY,
2002 (in press).
4. Matsushima T, Akabayashi A, Nishitateno K. The current status of bereavement follow-up in hospice andpalliative care in Japan. Palliative Medicine 16: 151158, 2002.
5. Akabayashi A. “To Intervene?”, “Misrepresenting
Research” (commentary). In Thomasma DC andKushner T eds., Ward Ethics, Cambridge University
Press, NY, pp. 244-247, 249-251, 2001.
6. Akabayashi A, Voltz R. Advance Directives in
Different Cultures. In Portenoy RK and Bruera E eds.,
Topics in Palliative Care vol. 5, Oxford University
Press, New York, NY, pp.107-122, 2001.
7. Lee JS, Kawakubo K, Sato H, Kobayashi Y, Haruna Y.
Relationship between total and regional bone miner-

al density and menopause state, body compositionand life style factors in overweight Japanese women.
Int J Obese 25: 880-886, 2001.
8. Lee JS, Kawakubo K, Kobayashi Y, Mori K, Kashihara
H, Tamura H. Effects of ten year body weight variability on cardiovascular risk factors in Japanese middle-aged men and women. Int J Obese 25: 10631067, 2001.
9. Akabayashi A, Fetters MD. Paying for informed consent. Journal of Medical Ethics 26: 3; 212-214, 2000.
10. Fukusaki C, Kawakubo K, Yamamoto Y. Assessmentof the primary effect of aging on heart rate variability
in humans. Clinical Autonomic Research 10: 123130, 2000.
11. Kashihara H, Ohno H, Tamura M, Kawakubo K,
Gunji A. Risk factors for ischemic heart disease in an
urban area of Japan: a case-control study in AMHTS.
Method Inform Med 39: 223-228, 2000.
12. Ishizaki T, Kobayashi Y, Kai I. Functional transitionsin instrumental activities of daily living among older
Japanese. Journal of Epidemiology 10: 249-254, 2000.
13. Akabayashi A, Kai I, Takemura H, Okazaki H. Truth
telling in the case of a pessimistic diagnosis in Japan.
The Lancet 354;1263, 1999.
14. Akabayashi A, Fetters MD, Elwyn TS. Family consent, communication, and advance directive for cancer disclosure: A Japanese case and discussion.
Journal of Medical Ethics 25 : 4; 296-301, 1999.
15. Miyasaka M, Akabayashi A, Kai I, Ohi G. An international survey of medical ethics curricula in Asia.
Journal of Medical Ethics 25: 6; 514-521, 1999.
16. Voltz R, Akabayashi A, Reese C, Ohi G, Sass HM.
Attitude of health care professionals towards clinicaldecisions in palliative care: A cross-cultural comparison. Journal of Clinical Ethics 10: 4; 309-315, 1999.
17. Akabayashi A. Transplantation from a Brain Dead
Donor in Japan. The Hastings Center Report MayJune 29(3): 48, 1999.
18. Haruna Y, Lee JS, Kawakubo K, Suzuki Y.
Cardiovascular reserve to orthostatic stress in active
living elders. J Gravit Physiol 6: 107-108, 1999.
19. Suzuki Y, Takenaka K, Haruna Y, Akima H,
Fukunaga T, Kawakubo K, Gunji A. Relatively shorter but intensive isotonic exercise in legs prevented
the decrease in maximal aerobic power after 20 daysof head down tilt bed rest in man. J Gravit Physiol 6:
117-118, 1999.
20. Oba M, Kawakubo K, Lee JS, Haruna Y, Suzuki Y,
and Gunji A. Degraded Diet Induced Thermogenesis
Induced by 20-days 6º Head-Down Bed Rest and the
Effect of Resistance Training. J Gravit Physiol 6: 14950, 1999.
21. Akabayashi A. Female circumcision - A health issue
or a human rights issue. Health Care Analysis 6 (1):55-58, 1998.
22. Takai A., Wongkhomthong S, Akabayashi A, Kai I,
Ohi G, Naka K. Correlation between history of contact with people living with HIV/AIDS(PWAs) and
Tolerant Attitudes toward HIV/AIDS and PWAs inrural Thailand. Int J STD and AIDS 9: 482-484, 1998.

309

23. Voltz R, Akabayashi A, Reese C, Ohi G, Sass HM.
End-of-life decisions and advance directives in palliative care: A cross-cultural survey of patients andhealth care professionals. Journal of Pain and
Symptom Management 16: 153-162, 1998.
24. Ohi G, Akabayashi A, Miyasaka M. Japan’s healthcare system as egalitarian: A brief analysis of its history and selected social systems. Health Care
Analysis 6 (2): 141-149, 1998.
25. Nishimura-Takahashi T, Akabayashi A, Kai I,
Cabigon J, Ohi G, Naka K. Social and Behavioral
Factors Associated with condom use among femalecommercial sex workers (CSWs) in Tarlac, thePhilippines. Environmental Health and Preventive
Medicine 2(4): 167-171, 1998.
26. Yanagibori R, Kondo K, Suzuki Y, Kawakubo K,
Iwamoto T, Itakura H, Gunji A. Effect of 20 days’ bed
rest on the reverse cholesterol transport system inhealthy young subjects. J Intern Med 243: 307-312,
1998.
27. Murakami T, Yamada N, Kawakubo K, Oku J,
Mizushima Y, Lino S, Sugimoto T. Lipoprotein
abnormalities in non-insulin dependent diabetic
patients with ischemic heart disease. Artery 22: 206232, 1997.
28. Akabayashi A. Finally done -- Japan’s final decision
on organ transplantation. The Hastings Center
Report Sep-Oct 47, 1997.
29. Akabayashi A. Japan’s parliament passes brain-deathlaw. The Lancet 349: 9069, 1895, 1997.
30. Voltz R, Akabayashi A, Reese C, Ohi G, Saas HM.
Organization and patients’ perception of palliative
care: A crosscultural comparison. Palliative Medicine
11: 351-357, 1997.
31. Gunji. A. Short review of human prolonged horizontal bed rest studies in Japan. J Gravit Physiol 4: S1-9,
1997.
32. Akabayashi A. The concept of happiness in oriental
thought and its significance in clinical medicine. In
Japanese and Western Bioethics , Philosophy &Medicine Series 54, Kluwer Academic Publishers,
Netherlands, pp.161-164, 1997.
33. Akabayashi A. Questions raised over release of sideeffects data in Japan. The Lancet 350: 9071, 124,
1997.
34. Sass H, Bonkovsky F, Akabayashi A, Kielstein R,
Olick RS. Advance Health Care Documents in
Multicultural Perspective. Ann. Rev. of Law andEthics. Band 4, 465-508, 1996.
35. Suzuki Y, Akima H, Igawa S, Fukunaga T, Kawakubo
K, Goto S, Makita Y, Gunji A. Decrease of bone mineral density and muscle and/or strength in the leg
during 20 days horizontal bed rest. J Gravit Physiol
3: 42-43, 1996.
36. Linnarsson D, Sundberg CJ, Tedner B, Haruna Y,
Karemaker JM, Antonutto G, Prampero PE. Bloodpressure and heart rate responses to sudden changesof gravity during exercise. Am.J.Physiol. (Heart Cir.
Physiol. 39): H2132-H2142, 1996.

310

37. Akabayashi A. Financing health care - A Japaneseperspective. Health Care Analysis 3: 123-125, 1995.
38. Akabayashi A. Open letter to those diet members
responsible for legislating Japanese organ transplantation. Health Care Analysis 3: 358-360, 1995.
39. Suzuki Y, Haruna Y, Kuriyama K, Kawakubo K, Igawa
S, Goto S, Makita Y, Gunji A. Effects of 20 days bed
rest on mechanical efficiency during upright cycling
and leg muscle mass in young males. J Gravit Physiol
2: 74-75, 1995.
40. Haruna Y, Suzuki Y, Yanagibori R, Kawakubo K,
Gunji A. Activated renin-angiotensin-aldosterone
system and its effects on cardiovascular system during 20-days horizontal bed rest. J Gravit Physiol 2:
27-28, 1995.
41. Miyashita S, Haruna Y, Suzuki Y, Gunji A. Effects ofposture on cardiorespiratory responses during mild
exercise. J. Phys. The. Sci. 7: 21-25, 1995.
42. Lee JS, Aoki K, Kawakubo K, Gunji A. A study on
indices of body fat distribution for screening for obesity. J Occup Health 37: 9-18,1995.
43. Jeong HS, Gunji A. The influence of system factors
upon the macro-economic efficiency of health care:
implications for the health policies of developing anddeveloped countries. Health Policy 27: 113-40,1994.
44. Koyano W, Hashimoto M, Gunji A, Fukawa T,
Shibata H. The social support system of the Japaneseelderly. Journal of Cross-Cultural Gerontology 9: 32333,1994.
45. Kawakubo K, Gunji A, Sakamoto S, Nakamura F.
Exercise-induced ST segment changes and coronary
risk factors in asymptomatic men. Jpn Heart J 35:
531-532, 1994.
46. Suzuki Y, Kashihara H, Takenaka K, Kawakubo K,
Makita Y, Goto S, Ikawa S, Gunji A. Effects of daily
mild supine exercise on physical performance after
20 days bed rest in young persons. Acta Astronautica
33: 101-11,1994.
47. Bonde-Petersen F, Suzuki Y, Kawakubo K, Gunji A.
Capillary filtration rate, venous compliance, andblood flow in arms and legs do not change during
bed rest for 20 days. J Grav Physiol 1: 27-8, 1994.
48. Suzuki Y, Murakami T, Kawakubo K, Haruna Y,
Takenaka K, Goto S, Makita Y, Igawa S, Gunji A.
Regional changes in muscle mass and strength following 20 days of bed rest, and the effects on orthostatic tolerance capacity in young subjects. J Grav
Physiol 1: 57-8,1994.
49. Akabayashi A, Morioka M. Ethical issues raised by
medical use of brain-dead bodies in the 1990s.
BioLaw II (48): S531-538, 1991.
50. Akabayashi A, Morioka M. Research on dead persons. Ann. Intern. Med. 111 (1): 89, 1989.

Department of Biostatistics /Department of Epidemiology
and Preventive Health Sciences
Outline and Research Objectives
These two departments were established from the department of Biostatistics andEpidemiology in 1996 according to the reorganization of the Graduate School. They are now
jointly managed under the supervision of Prof. Ohashi, who moved to the School in 1990
from the University Hospital Computer Center, the University of Tokyo Hospital. When
Prof. Ohashi moved to the School, the name of the department was ‘Epidemiology’ and thename was changed into the above ‘Department of Biostatistics and Epidemiology’ in 1992
according to the reorganization of the undergraduate and graduate schools and their educational systems. This department was the first official department of biostatistics in Japanese
universities, which shows this interdisciplinary and indispensable specialty field required
for medical research has been neglected in Japanese university educational system. Now
there are three departments of biostatistics; the two others are in Kitasato University (from1999) and Kyoto University (from 2000), respectively. Our department has, as the firstdepartment which can provide comprehensive educational courses in epidemiology and biostatistics from the under-graduate level, brought up and sent many biostatisticians and theoretical epidemiologists to research institutions including Kyoto University, the Ministry of
Health, Labor and Welfare and the National Cancer Center as well as pharmaceutical industry.
(1) Development of statistical-analytical methodology required for medical research including genome data analysis (such as microarray data)(2) Development of methodology of clinical trials including data management(3) Collaborative epidemiological research (JALS Study described later)(4) QOL research(5) Outcome research, pharmacoeconomical and/or pharmacoepidemiological research
The last research project is now conducted in collaboration with the department ofPharmacoepidemiology (established in 1993 with the main support from our department)
and the department of Pharmacoeconomics in the School of Pharmacy. The partnershipwith the Clinical Bioinformatics Educational Unit, which was just established in 2002, will
deepen and expand our research activities especially in the field of clinical epidemiology.

Faculty and Students
Professor and Chair Yasuo Ohashi Ph.D. (1990-)
Associate .................................2
Postdoctoral Fellow ................3
Graduate student7 (doctor) 8 (master)
Research student...................12
Secretary .................................3

Past Research and Major Accomplishments(number in the parenthesis is the paper inthe selected list)
A new methodology of random allocation, the centralized dynamic balancing, was firstly introduced by
Ohashi (in 1988 and (1)) into Japan and implemented
in many clinical fields rather than oncology, the original application field. The possibility of utilizing Web(internet) system for registration/randomization was
firstly (in the world) discussed in (21) and implemented in the UMIN (University Medical Information
Network: one of the largest medical network systemmanaged by the University Hospital Computer
Center). As a statistician, Ohashi participated into
many clinical trials, especially large-scale long-termclinical trials which had been very rare in Japan until
1990 and contributed to the design, conduct andanalysis. Clinical areas include: oncology (37), hematology (25), organ-transplantation, bone-marrowtransplantation (45), infectious diseases (8,38), diabetes, cardiovascular diseases (48), hypercholesterolemia (44), cerebrovascular diseases (10,36), kidney diseases (28), neurological disorders and urology(9,32). These collaborative studies stimulated new
methodological developments in design and analysisof clinical trials. Examples include:311

Fig.1 Clinical Trial Research Model・ Bayesian dose-escalation design in early cancer
clinical trials (29,50)
・ Drug randomization and deliverly system using
network (35)・ Analysis method for investigating institutional differences (27,33,34,47)
・ Analysis method of recurrent events (31)・ Analysis method of bivariate longitudinal data
(23,41)
Consultation is a major mission of biostatisticiansin medical institutions and contributes to new findings in each research area as well as confirming usefulness of statistical methodology and education ofyoung statisticians. Illustrative examples we experienced include:・ Application of time-dependent covariate analysis(2,4,6)・ Application of kernel smoothing (30)・ Application of cluster analysis (5)Ohashi and his colleague conducted several metaanalyses of Japanese original oncology drugs, which
had been widely used in the surgery adjuvant setting
and criticized due to lack of clinical evidence, and provided the data for evaluating the risk-benefit ratio(42). Pharmacoepidemiology is a new active field
explored in collaboration with the department ofPharmacoepidemiology. Besides specific reserches(15,39), we have established the original adverseevents monitoring system (J-PEM) with participationof pharmacists and published the results of pilot studies (40,49). Some pure epidemiological researches
have been conducted by Ohashi and colleagues(19,46), and they are highly evaluated for their rigorous methodology. These experiences lead to a newresearch area, a prospective collaborative meta-analy-

312

sis (JALS) of Japanese representative cohort studies
which is described later.
QOL research is an interdisciplinary area which
requires collaboration with not only care providers
but also other specialists such as psychologists andstatisticians. Ohashi started pioneering works inJapanese oncology area (12,43) as a statistician and arepreparing several papers based on the results of largescale clinical trials in breast cancer and non-small-celllung cancer.

Current Research and Future Prospects
The research objectives described in the earlier
section are pursued in collaboration with inside andoutside researchers as in the past; the recent topics
includes:・ Analysis method for multiple (different type)
events・ Joint modeling of longitudinal data and events・ Analysis method for determining the racial difference in drug responses
・ Method of sample size re-estimation in clinical trials
・ Method of cluster randomization in clinical trials
In addition, new research models emerged recently.
In 2001, Ohashi established a NPO titled Japan
Clinical Research Support Unit (J-CRSU) as a chair-person for・ Support for investigator-initiated clinical researchthrough statistical analysis, data management andquality assurance
・ Educational support of clinical research methodology and biostatistics, especially education of clinical research coordinators (CRC; research nurses)
・ Support for education and dissemination of medical writing and he also started a model projectFig.2 JALS participating cohorts

called Comprehensive Support Project forOncology Research-Breast Cancer (CSPOR-BC) in
the Public Health Research Foundation for・ Planning and conduct of investigator-initiated clinical trials for breast cancer patients
・ QOL research in oncology
・ Education of CRC
・ Information support for breast cancer patients andresearchers/CRC
The research model is described in Fig.1, where
quality control is conducted through the collaboration
between data center located in the NPO and CRC dispatched from site management organizations (SMO)or CRC hired in each institution. For increasing efficacy (reducing cost) and keeping security, remote data
entry system utilizing UMIN and private virtual network system is designed. Three clinical trials on thisscheme are ongoing or will be started by the end of
2002 (2 breast cancer trials (1200 and 2500 patients,
respectively) supported by CSPOR-BC and 1 osteoporosis trial (1800 patients) supported by the similar
project called A-TOP).
One big epidemiological research project called
Japanese Arteriosclerosis Longitudinal Study (JALS)
has started from 2001 supported by the Japanese
Arteriosclerosis Prevention Fund. The objective of
JALS is to conduct a prospective meta-analysis collecting the base-line and follow-up data from individuallyconducted Japanese cohort studies (including thefamous Hisayama cohort) and estimate the gender-age
specific incidence rates of cardiovascular and cerebrovascular events as well as quantify the influence ofvarious risk factors. The operation office is located in
the NPO and we have conducted the standardization

procedures including standardization of blood-pressure measurements, lipid measurements and developments of the common questionnaires for nutrition
intake and physical activities. At present (November,
2002), the cohort size is expected to be 90,000(regional cohorts) and 25,000 (job-office cohorts) as
shown in Fig.2.
All department staffs and students are involved in
the above research activities and these models will
provide new opportunities of collaboration and education not only for inside persons but also outside
researchers and students as well as industry persons.
Recent research topics conducted in these activities include:
・ Development of analytical methodology required
for meta-analysis
・ Validation of physical activity questionnaire using
a newly developed instrument measuring acceleration every 4 seconds
・ QOL and neurotoxicity by chemotherapy in breast
cancer patients・ Epidemiology of postmenopausal symptoms anddepression in breast cancer patients
A variety of research fields are expected to emerge
from mutual stimulation and enlightenment.

Research Grants
11 grant projects from the Ministry of Health, Laborand Welfare and 1 grant project from the Ministry of
Education and Science are ongoing in 2002 including:(1) Establishment of the tailor-made strategy for osteoporosis(2) Mild hypothermia therapy for severe head injuries inhuman: a multicenter randomized control trial

313

(3) Establishment of novel treatment strategy for breast
cancer using cDNA array technology(4) Clinical development of new modality of allogeneic
hematopoietic stem cell transplantation with areduced-intensity regime(5) Study on establishment of clinical trial system of
good quality through cooperative group mechanism

Select Publications (* with reprints)
1. Ohashi Y : Randomization in cancer clinical trials :Permutation test and development of a computer
program. Environmental Health Perspectives; 87: 137., 1990.
2. Funatsu H, Yamashita H, Ohashi Y, Ishigaki T :Correlation between progression of diabetic retinopathy and blood gulcose control. Japanese J.
Ophthalmology; 35: 306-16., 1991.
3. Shiina S, Fujino H, Uta Y. Ohashi Y et al. :Relationship of HbsAg subtypes with HbeAg/antiHbe status and chronic liver disease. Part I: Analysisof 1744 HbsAg carriers. The American J.
Gastroenterology; 86: 866-71., 1991.
4. Funatsu H, Yamashita H, Ohashi Y, Ishigaki T : Effectof rapid glycemic control on progression of diabetic
retinopathy. Japanese J. Ophthalmology; 36: 356-67.,
1992.
5. Suzuki Y, Araie M, Ohashi Y : Sectorization of the central 30゜visual field in glaucoma. Ophthalmology;
100(1): 69-75., 1993.
6*. Yoshimoto M, Sakamoto G, Ohashi Y : Time dependency of the influence of prognostic factors on relapse
in breast cancer. Cancer; 72(10): 2993-3001., 1993.
7. Yamada N, Gotoda T, Yoshinaga H, Ohashi Y et al :Increased risk factors for coronary artery disease inJapanese subjects with hyperinsulinemia or glucose
intolerance. Diabetes Care; 17(2): 107-14., 1994.
8. Morikawa Y, Ohashi Y, Harada K, Asai T et al : A multicenter, randomized, controlled trial of intravenous
gamma globulin therapy in children with acute
Kawasaki disease. Acta Paediatrica Japonica; 36: 34754., 1994.
9. Isaka S, Shimazaki J, Akimoto S, Ohashi Y et al : A
prospective randomized trial for treating stages B2and C prostate cancer : Radical surgery or irradiation
with neoadjuvant endocrine therapy. Japanese J.
Clinical Oncology; 24(4): 218-23., 1994.
10. Sasaki T, Ohta T, Kikuchi H, Ohashi Y et al : A phase
II clinical trial of recombinant human tissue-type
plasminogen activator against cerebral vasospasm
after aneurysmal subarachnoid hemorrhage.
Neurosurgery; 35(4): 597-605.
11. Matsumoto T, Yamada N, Ohashi Y, Kikuchi M :Coronary heart disease mortality is actually low indiabetic Japanese by direct comparison with the
Joslin cohort. Diabetes Care; 17(9): 1062-3., 1994.
12. Yasue M, Sakamoto J, Morimoto T, Ohashi Y et al :Evaluation of the effect of pancreatic resection inadvanced pancreatic cancer with special reference314

using hospital-free survival as a measure of quality oflife. Japanese J. Clinical Oncology; 25: 37-45., 1995.
13. De Mulder PHM, Williams R, Ohashi Y, Blumenstein
BA et al : The Design of clinical trials for new therapies in bladder cancer: Is it time for new strategies?
International J. Urology; 2(Suppl2): 113-6., 1995.
14. Tanimoto K, Nakano K, Kano S, Ohashi Y et al :Classification criteria for polymyositis and dermatomyositis. The Journal of Rheumatology; 22(4): 66874., 1995.
15. Matsuyama Y, Ohashi Y, Uchino H, Takaku F et al :Vesnarinone-induced WBC Disorders in Japan.
Pharmacoepidemiology and Drug Safety; 5: 87-93.,
1996.
16. Sakamoto J, Teramukai S, Koike A, Ohashi Y et al :Prognostic value of preoperative Immunosuppressive
Acidic Protein in patients with gastric carcinoma.
Cancer; 77(11): 2206-12., 1996.
17. Homma Y, Kawabe K, Tsukamoto T, Yamaguchi, O,
Ohashi Y et al : Estimate criteria for diagnosis andseverity in benign prostatic hyperplasia.
International J. of Urology; 3: 261-6., 1996.
18. Sato K, Nohtomi K, Shizume K, Ohashi Y et al : 17
β-Estradiol increases calcium content in fetal mouseparietal bones cultured in serum-free medium only at
physiological concentrations. Bone; 19(3) :213-21.,
1996.
19. Honda H, Shimizu Y, Misumi K, Niimi M, Ohashi Y :Cumulative incidence and prevalence of childhood
autism in children in Japan. British Journal
Psychiatry; 169: 228-35., 1996.
20. Ohashi Y, Izumoto T : Comparison of intra-subject
variability for different formulation by crossover trials. F.I.P.Bio-International`96;113-8., 1996.
21*. Kiuchi T, Konishi M, Bandai Y, Kosuge T, Ohashi Y: A World Wide Web-based user interface for a data
management system for use in multi-institutional
clinical trials-Development and experimental operation of an automated patient registration and random allocation system. Controlled Clinical Trials; 17:476-93., 1996.
22. Kim−Motoyama H, Yasuda K, Yamaguchi T,
Yamada N, Ohashi Y : A mutation of the β3−adrenergic receptor is associated with visceral obesity but decreased serum triglyceride. Diabetologia; 40:469-72., 1997.
23*. Matsuyama Y, Ohashi Y : Mixed models for bivariate response repeated measures data using Gibbs
sampling. Statistics in Medicine 1997; 16:1587-601.
24. Fukuda H, Ohashi Y: A guideline for reporting resultsof statistical analysis in Japanese Journal of ClinicalOncology. Japanese J. of Clinical Oncology 1997;
27(3): 121-7., 1997.
25. Ohno R, Miyawaki S, Hatake K, Kuriyama K, Ohashi
Y et al : Human urinary macrophage colony - stimulating factor reduces the incidence and duration offebrile neutropenia and shortens the period required
to finish three courses of intensive consolidation
therapy in acute myeloid leukemia: A double-blindcontrolled study. J. of Clinical Oncology 1997; 15(8):2954-65., 1997.
26. Hukuhara S,Tanabe N,Sato K,Kurokawa K,Ohashi Y :Good Clinical Practice in Japan before and after ICH:
Problems and potential impacts on clinical trials andmedical practice. International J. of Pharmaceutical
Medicine; 11: 147-53., 1997.
27*. Matsuyama Y, Sakamoto J, Ohashi Y : A Bayesianhierarchical survival model for the institutional
effects in a multi-centre cancer clinical trial. Statistics
in Medicine; 17: 1893-908., 1998.
28. Akiba T, Marumo F, Owada A, Ohashi Y et al :Controlled trial of falecalcitriol versus alfacalcidol in
suppression of parathyroid hormone in hemodialysis
patients with secondary hyperparathyroidism.
American J. of Kidney Diseases; 32(2): 238-46., 1998.
29. Furuse K, Kawahara M, Nishiwaki Y, Ohashi Y :Phase I/II study of vinorelbine, mitomycin and cisplatin for stage IIIB or IV non-small-cell lung cancer.
Journal of Clinical Oncology ; 17-10: 3195-200., 1999.
30*. Hinotsu S, Akaza H, Kotake T, Ohashi Y :Intravesical chemotherapy for maximum prophylaxisof new early phase superficial bladder carcinoma
treated by transurethral resection: A combined
analysis of trials by the Japanese Urological Cancer
Research Groupusing smoothed hazard function.
Cancer; 86: 1818-26., 1999.
31*. Matsui S, Ohashi Y : Analysis of recurrent events:
Application to a clinical trial of colony stimulating
factor with the endpoint of febrile neutropenia.
Statistics in Medicine; 18: 2409-20., 1999.
32. Akakura K, Isaka S, Akimoto S, Ohashi Y et al :Long-term results of a randomized trial for the treatment of stages B2 and C prostate cancer : Radical
prostatectomy versus external beam radiation therapy with a common endocrine therapy in both modalities. Urology; 54-2 : 313-8., 1999.
33. Skamoto J, Mtsuyama Y, Ohashi Y: An analysis ofinstitutional effects in a multicenter cancer clinical
trials- Is it also plausible from the clinicians’ point ofview-. Japan Journal of Clinical Oncology;29-8:403-5.,
1999.
34. Yamaguchi T, Ohashi Y : Investigating centre effects
in a multi-centre clinical trials of superficial bladdercancer. Statistics in Medicine; 18-15: 1961-71., 1999.
35*. Gotoh F, Ohashi Y and the Cilostazol Stroke
Prevention Study group: Design and organization of
the Cilostazol Stroke Prevention Study. J. of Stroke
and Cerebro- vascular Diseases; 9: 36-44., 2000.
36. Gotoh F, Tohgi H, Hirai S, Ohashi Y et al : Cilostazol
Stroke Prevention Study : A Placebo-controlled double-blind trial for secondary prevention of cerebral
infarction. J. of Stroke and Cerebrovascular Diseases;
9:147-57., 2000.
37. Takayama T, Sekine T, Makuuchi M, Ohashi Y et al :Adoptive immunotherapy to lower postsurgicalrecurrence rates of hepatocellular carcinoma: a randomised trial. The Lancet; 356: 802-7., 2000.
38. Morikawa Y, Harada K, Asai T, Ohashi Y et al :Coronary risks after high-dose γ-globulin in chil-dren with Kawasaki disease. Pediatrics International;
42: 464-9., 2000.
39*. Matsuyama Y, Tominaga T, Nomura Y, Ohashi Y etal : Second cancers after adjuvant tamoxifen therapy
for breast cancer in Japan. Annals of Oncology; 11:1537-43., 2000.
40*. Kubota K, Kawabe E, Hinotsu S, Ohashi Y et al :Pilot study of Prescription-Event Monitoring in Japan
comparing troglitazone with alternative oral hypoglycemics. Eur. J. Clin. Pharmacology; 56:831-8.,
2001.
41. Toyoda I, Matsuyama Y, Ohashi Y : Estimation andcomparison of rates of change in repeated-measures
studies with planned dropout. Controlled Clinical
Trials; 22(6): 620-38., 2001.
42. Sakamoto J. Teramukai S. Watanabe Y. Ohashi Y etal : Meta-analysis of adjuvant immunochemotherapy
using OK-432 in patients with resected non-smallcell lung cancer. Journal of Immunotherapy; 24(3):250-6., 2001.
43. Fumimoto H. Kobayashi K. Chang C H. Ohashi Y. et
al : Cross-cultural validation of an international questionnaire, the general measure of the Functional
Assessment of Cancer Therapy Scale (FACT-G) forJapanese. Quality of Life Research; 10: 701-9., 2001.
44. Ito H. Ouchi Y. Saito Y. Ohashi Y. et al : A comparison of low versus standard dose pravastatin therepy
for the prevention of cardiovascular events in theelderly: The Pravastatin Anti-atherosclerosis Trial in
the Elderly (PATE). Journal of Atherosclerosis andThrombosis; 8(2): 33-44., 2001.
45. Hiraoka A. Okamoto S. Moriyama Y. Ohashi Y. et al :Phase III study comparing tacrolimus (FK506) withcyclosporine for graft-versus-host disease prophylaxis after allogeneic bone marrow transplantation.
Bone Marrow Transplantation; 28: 181-5., 2001.
46*. Ohashi Y, Nakai S, Tsukamoto T, Masumori N et al: Habitual intake of lactic acid bacteria and risk
reduction of bladder cancer. Urologia Internationalis;
68: 273-80., 2002.
47. Yamaguchi T, Ohashi Y, Matsuyama Y : Proportional
hazards models with random effects to examine centre effects in multicentre cancer clinical trials.
Statistical Methods in Medical Research; 11: 221-36.,
2002.
48. The EPOCH Study Group : Effects of pimobendan on
adverse cardiac events and physical activities inpatients with mild to moderate chronic heart failure:
The effects of pimobendan on chronic heart failure
study (EPOCH Study) Circulation J.; 66: 149-57.,
2002.
49. Samizo K, Kawabe E, Hinotsu S, Ohashi Y et al :Comparison of losartan with ACE inhibitors anddihydropyridine calcium channel antagonists. A pilot
study of Prescription-Event Monitoring in Japan.
Drug Safety; 25: 811-21., 2002.
50. Ishizuka N, Ohashi Y : The continual reassessment
method and its applications: a Bayesian methodology
for phase I cancer clinical trials. Statistics in
Medicine; 20: 2661-81., 2001.

315

Department ofAdvanced Clinical Nursing /Department of
Nursing Administration
Outline and Research Objectives
The origins of the Departments of Advanced Clinical Nursing and Nursing
Administration can be tracked down to the Department of Fundamental Nursing, one offour nursing departments in the School of Health Care and Nursing. In 1953, the School of
Health Care and Nursing was instituted at the Faculty of Medicine, the University of Tokyo,
as the second 4-year baccalaureate nursing program in Japan. In 1965, the school was reorganized and renamed as the School of Health Sciences, with a focus on health sciences education. Accordingly, the four nursing departments were integrated into one department, theDepartment of Nursing, which became primarily responsible for undergraduate nursing education at the school until 1992. In that year the school was again, reorganized and renamed
as the School of Health Sciences and Nursing, taking on the nation-wide momentum forenhancing baccalaureate programs in nursing. Three departments, including adult healthand nursing, family nursing, and community health nursing, were instituted in addition to
the Department of Fundamental Nursing (formerly the Department of Nursing). In 1997, as
the result of the shift to the chair system of the Graduate School of Medicine, theDepartment of Fundamental Nursing was reorganized into two graduate departments—the
Department of Advanced Clinical Nursing and the Department of Nursing
Administration—.
Currently, these two departments are also responsible for undergraduate education in
the following subjects: Fundamental Nursing (introductory lectures, skills laboratory, andpracticum; 10 credits in total); First Aid and Cardiopulmonary Resuscitation (1 credit);
Nursing Research (2 credits); and Nursing Administration (lecture and practicum, 2 credits).
The major research activities of the two departments include: studies on the links between
nursing quality and patient safety and outcomes; studies on nursing management issues;
the development and validation of nursing quality measurements; and physiological studies
on human body responses to psychophysiological stress (e.g., patients’ responses to invasive
treatment, family caregivers’ responses to nighttime caregiving activities).

Faculty, Staff, and Students
Professor and Chair

Katsuya KANDA, R.N., PhD. (since
2002)
Associate .................................2
Graduate student ....................7
Research student.....................4
Secretary .................................2

Past Research and Major Accomplishments
The author has primarily been conducting studies
on nursing administration, especially on the issue ofnursing quality and care delivery management.
Secondly, the author has been conducting physiological studies on human body responses to psychophysiological stress and stimulus, such as invasive treat-316

ment or intensive caregiving activities.
The author’s initial research involvement in thearea of nursing administration was a study of nursing
home staffing in the United States of America (USA).
In 1983, as part of the national strategies to control
the ever-increasing medical expenditures in the
Medicare reimbursement system, the federal government of the USA introduced the prospective payment
system (PPS), utilizing the diagnosis related groups(DRGs) as the payment basis. As a result of the PPS
introduction, elderly patients who were not fully
recovered from their illnesses were transferred from
hospitals to nursing homes. However, the tight budget situation brought about reductions in the numberof nursing home staff and the quality of nursing,despite ever-increasing disease severity and careneeds of nursing home residents. The author and col-leagues analyzed the time-series data of multiple nursing homes obtained from a health data center in thestate of Pennsylvania, and verified the above-mentioned issue of understaffed nursing homes.
In Japan, the author conducted a series of studies
on the links between nurse staffing and patient safety
and outcomes. The primary factor affecting the quality of nursing is the nurse-patient ratio. The claim thatthe understaffing of nurses brings about a too busy
working environment for nurses to pay attentions to
the details of patient care (which may cause unfavorable consequences to patients) has been an unspoken
agreement among nurses for many years. The average
length of hospital stay is one of the patient outcome
indices that reflect the quality of healthcare and nursing. The author obtained permission from the
Ministry of Health and Welfare (MHW) to use thedata of “Hospital Reports” surveyed by the MHW, andanalyzed the relationship between the number of“nursing staff” (including licensed practical nurses andnurses’ aids) assigned per bed and the averagepatients’ length of hospital stay. The data obtained
from approximately 7,000 general hospitals were analyzed in this study. The average length of stay amonghospitals with a high “nursing staff”-to-bed ratio wasshorter than that of hospitals with a low “nursing
staff”-to-bed ratio. The average length of stay amonghospitals with a high ratio of registered nurses (RNs)
to “nursing staff” tended to be shorter than that of
hospitals with a low RNs-to-“nursing staff” ratio.
Similar results were confirmed after variables related
to hospital characteristics (which are known to have astrong link to patient characteristics) were adjusted.
The author then obtained permission from theManagement and Coordination Agency to use thedata of “Patient Survey” and “Survey of Medical Care
Institutions” surveyed by the MHW, and analyzed therelationship between the nurse staffing level approvedon the nursing fee schedule of health insurance revenue and the incidence of adverse events, such ascomplications. The “Patient Survey” data have twovariables related to diagnostic classifications (the primary diagnosis and the secondary diagnosis, if applicable). Two nurse researchers with advanced clinicalexpertise independently evaluated the incidence ofadverse events from approximately 295,000 in-hospital patients’ survey data. Discrepancies between thetwo nurse researchers’ evaluations were discussed andresolved by incorporating a third nurse researcher’s
independent evaluation. The result of this adverse
event analysis was appalling: the decreased number ofnurses per patient was associated with an increased
incidence of adverse events; especially, the incidenceof adverse events was different between hospitals
with a patient-to-nurse ratio of 2.5-to-1 and hospitals
with a ratio of 3-to-1.

Japan has fewer numbers of registered nurses
assigned to patients in acute care hospitals comparedto other developed countries. A large number of registered nurses per patient does not necessarily bringabout better quality of care; however, a small numberof registered nurses per patient never brings about
better quality of care. The author maintains that an
adequate number of registered nurses assigned per
patient is a “necessity” of better quality of nursing.
In relation to studies of care management, theauthor and colleagues actually developed and implemented a critical pathway for patients with abdominal aortic aneurysms undergoing surgery. The authors
measured the effects of the critical pathway before
and after its introduction to a vascular surgical careunit at the University of Tokyo Hospital. A criticalpathway is one of the patient care management tools.
Multidisciplinary members involved in patient careneed to participate in the process of critical pathway
development, making suggestions and modifications
on pathways from their clinical expertise. Nurses
should clarify as to when and for which components
of care they are accountable. Therefore, a study ofcritical pathway implementation is also a study ofnursing care itself. Additionally, critical pathways arehighly effective for those patients with high-risk diseases who tend to be vulnerable to multiple complications, and without adequate care management, theytend to have delayed recovery and discharge. We disseminated the study results to nurses and educatedthem, emphasizing that critical pathways are usefulfor high-risk patients, and that a comprehensive discussion among multidisciplinary team members isnecessary during the process of pathway development.
One of the most serious nursing administrative
issues involves the frequent uses and their side effectsof physical restraint on patients. Regarding physiological studies, the author and colleagues studied theeffects of physical restraint on patients via electroencephalography. Elderly healthy volunteers participating in this experimental study with a crossover design
received loose physical restraints. Their electroencephalography data showed deviations from normal
patterns, despite the fact that they rarely reported
subjective discomfort. Nursing research also encompasses patient family members as study subjects. In
order to contribute to the knowledge of interventions(the development of supporting systems) for nighttime family caregivers at home, the authors studied
the sleep patterns of female family caregivers who
were routinely providing nighttime care to bed-ridden
elderly individuals. The authors compared their sleep
patterns via electroencephalography data with thoseof an age-matched healthy control group. The study
results confirmed that the nighttime family caregiver317

group had less slow-wave sleep (deeper sleep stages)
than did the control non-caregiver group.

Current Research
A nation-wide healthcare reform, including systematic renovations to the functions of healthcare
facilities, is currently underway in Japan in order toimprove the efficacy of healthcare delivery systems.
Given this situation, healthcare facilities are beingclassified into two levels: acute care and long-termcare facilities. Nurses at long-term care settings needto assume a role different from that of acute care settings. For example, a high proportion of physicallydependent or mentally impaired residents (as opposed
to residents with medical needs), and a high proportion of care workers who are responsible for residents’personal care and assistance in their daily life activities are characteristic to long-term care settings. The
authors have just completed part of a survey involving
nurses’ roles, clinical expertise, job assignment andsharing in relation to care workers, and staffing levels
necessary to maintain a good quality of care at longterm care settings, such as health services facilities forthe elderly or sanatorium-type wards of hospitals.
The history of research on nursing care quality andthe methods of quality evaluation are very new toJapan; the availability of such data is very limited. The
authors are conducting studies to identify and establish patient outcome indicators that have: (1) known
empirical or theoretical strong links to the quality ofnursing; (2) reliable and accurate information widely
available to the majority of facilities; and (3) ease ofmeasurement or ease of access to information sources.
The authors are also attempting to standardize datacollection methods of such quality indicators. The
authors conducted a pilot study at a facility, identified
quality indicators meeting the above-mentioned criteria, and developed a trial version of standardized datacollection methods. A follow-up study at multiple
sites is underway in order to validate the utility ofthese indicators and methods.
Regarding studies on critical pathways, theauthors are currently conducting a study to examine
the utility of critical pathways in rehabilitation settings in addition to acute care settings. Specifically,
the authors are conducting a study of critical pathways for patients with hip fractures undergoing surgery, comparing the data from multiple sites regarding
the entire process of introduction and revisions ofcritical pathways, and the efficacy of the process.
Regarding studies of physical restraint, the authors
are examining an application of alternative and complimentary therapies that are recently attracting public attention as a method of alleviating psychologicalstress. When physical restraints are unavoidable with318

an understanding of their risks and unfavorable
effects, can we alleviate the psychosomatic distress ofpatients who are under restraint by using aroma therapy? We conducted an experimental study with a control group in order to measure the relaxing effects oflavender aroma on individuals under physicalrestraint. Currently, we are analyzing the electroencephalography and electrocardiography data.
Additionally, we are conducting a study to compare
the effects of the chair-sitting position (with the upper
body upright and detached from the back of a chair)and of the wheelchair-reclining position (with theupper body reclined on the back of a wheelchair) on
activities in the autonomous nervous system. Part ofthe literature and practice guidelines suggest the useof the chair-sitting position instead of the widely andfrequently used wheelchair-reclining position for
physically dependent or mentally impaired elderly
individuals. Some evidence supports the notion thatthe wheelchair-reclining position, given that the upper
body is totally reclined onto the back of the wheelchair, may decrease their ability to maintain theirbody position, or may decrease activities in the autonomic nervous system (as well as mental alertness).

Future Prospects
First, the long-term objective of research on nursing administration includes the development and validation of standardized methods and measurements(indices) of patient outcomes related to the quality ofnursing. A nation-wide database of nursing quality
indicators should be developed, starting off withinviting a certain number of hospitals willing to participate in the study. Continuous quality improvement
in healthcare and nursing involves organizational
efforts to identify their own national rankings and torenovate themselves through system-wide learningand changing. Such efforts of national database development and benchmarking would enable comparisonsamong organizations and would provide opportunities
for them to learn from top-ranked organizations.
When the nursing profession establishes such anational database of quality indicators, the author
believes that this profession would be able to takepolitical action and make suggestions with convincing
evidence to the government regarding concrete strategies for improving the quality of nursing. The author
would also like to conduct studies on the reliability
and sensitivity of quality indicators in order toincrease the validity of the comparisons.
Secondly, nursing research must provide convincing evidence that nursing can contribute to the healthpromotion of individuals/family/community (or the
optimization of their health experiences), so that theefficacy of nursing is reasonably recognized by thepublic. The effectiveness of nursing, including
patients’ psychophysiological responses to care, tended to be addressed more by anecdotal experiences
than by scientific studies providing evidence.
“Evidence-based nursing” is currently a widely-usedcatch phrase in Japan; however, the evidence regarding the effectiveness of care is still very weak. The
author believes that the Departments of Advanced
Clinical Nursing and Nursing Administration, which
are responsible for the fundamentals of nursing andskills laboratories in undergraduate education, arealso responsible for conducting studies on the efficacy
of nursing. With respect to critical pathways, many
retrospective studies have been conducted so far bynurse researchers in Japan. The author is planning toconduct a prospective and comparative study on theeffectiveness of critical pathways in order to obtain
more reliable and convincing evidence than that previously obtained. The author believes that randomized comparative studies from multiple sites are necessary to conduct in the future.
Finally, nursing informatics is one of the areas ofresearch and education still underdeveloped in thesedepartments. The author strongly believes that
research and educational programs of nursing informatics need to be developed in the future because of
the following reasons: (1) The implementation of abetter quality of nursing and an evaluation on its outcomes involves the development of evaluation criteria
and standards on the performance and quality ofnursing, and the actual implementation of evaluation
measures and procedures, thus ensuring accurate andhigh-quality information on healthcare and its administration; and (2) The study of nursing informatics
lays the foundation of practice, administration, education and research in nursing. Informatics in nursing
provides meaningful links among these areas and promotes development in each of these areas of nursing.

Research Grants
1. Principal Investigator: Physical Restraint of theElderly in Institutional Care Setting and Alternatives
to Restraint Use. Grant-in-Aid for Scientific Research,funded by the Ministry of Education, Science andCulture, 1996-1997.
2. Principal Investigator: Economic Evaluation of
Nursing Services. Grant-in-Aid for Health Sciences
Research, funded by the Ministry of Health andWelfare, 1996-1998.
3. Principal Investigator: Nurse Staffing and Nurses’ Job
by Type of Healthcare Facility. Grant-in-Aid for
Health Sciences Research, funded by the Ministry of
Health and Welfare, 1999-2001.
4. Principal Investigator: Effect Verification of
Development and Implementation of Critical
Pathway for Hip Fracture. Funded by JapaneseNursing Association, Project of Promotion of
Infrastructure Improvement for Nursing Policy
Planning, 2001.

Select Publications
1. Minamisawa H, Takeo K, Kanda K. Body positions
and heart rate: focusing on patient’s daily life.
Nursing 33(12): 44-51, 1981. (in Japanese)
2. Tagami Y, Hatano H, Hashimoto M, Kanda K, Tanaka
T. A study on the distribution of medical facilities insmall cities. Hospital Administration 19: 257-284,
1982. (in Japanese)
3. Takeo K, Minamisawa H, Kanda K, Hasegawa S: Heartrates during the daily activity of “Zen” priests.
Journal of Human Ergology 13: 83-87, 1984.
4. Kanda K. Promotion of home care and visiting nursing. The Tokyo Journal of Medical Science 95: 387390, 1988. (in Japanese)
5. Takeo K, Minamisawa H, Kanda K, Tukamoto M,
Kimura K, Mori Y, Ishida M, Watanuki S. Daily activity and food intake in patients with senile dementia.
Annual Report of Sasakawa Health Science
Foundation, 5(2), 103-112, 1989. (in Japanese)
6. Kanda K, Minamisawa H. How do nurse practitioners
use their time? In “The Improvement of the Quality
of Nursing Care by Introducing Primary Nursing,”
Report of Grant-in-Aid for Scientific Research from
the Ministry of Education, Science and Culture (pp.
65-82), 1990. (in Japanese)
7. Kanda K, Mezey M: Registered nurse staffing in
Pennsylvania nursing homes: Comparison before andafter implementation of Medicare’s Prospective
Payment System. Gerontologist 31: 318-324, 1991.
8. Kanda K, Maitoko M, Shimada Y, Takenawa N,
Awaya N, Mitoh M. How much are nurses performing other health care professionals’ duties in hospitals? Japanese Journal of Nursing Administration 1:
371-376, 1991. (in Japanese)
9. Kanda K, Takahashi M, Fujieda C. Nurse staffing at
outpatient clinics. Japanese Journal of Nursing
Administration 2: 55-61, 1992. (in Japanese)
10. Kanda K, Takahashi M, Fujieda C. Activities andfunctions performed by nurses in ambulatory caresetting. Japanese Journal of Nursing Administration
2: 179-185, 1992. (in Japanese)
11. Fujieda C, Takahashi M, Bojo N, Miyamoto A, Kanda
K. A survey on the levels of staffing by nurses in
hospitals. Japanese Journal of Nursing
Administration 2: 307-311, 1992. (in Japanese)
12. Kanda K, Fujieda C, Takahashi M, Bojo N, Miyamoto
A. Nurse staffing mechanism with relevance to casemix of patients. Japanese Journal of Nursing
Administration 2: 363-370, 1992. (in Japanese)
13. Kanda K, Takahashi M, Fujieda C, Bojo N, Miyamoto
A. Actual condition of hospital structure and equipment influencing nursing service. Japanese Journal of
Nursing Administration 3: 52-55, 1993. (in Japanese)
14. Kanda K. Trend of registered nurse staffing in nursing homes. Report of Research Activities by319

Sasakawa Health Science Fellowship Grantees, 7-17,
1993. (in Japanese)
15. Maitoko M, Kanda K. Factors effecting medication
errors. Journal of Japan Academy of Nursing Science
13(2): 29-36, 1993. (in Japanese)
16. Kanda K, Kusakari J, Awaya N. A study on the characteristics and length of employment of nonrotatingshift night nurses. Hospital Administration 30: 215222, 1993. (in Japanese)
17. Mitoh T, Takeo K, Minamisawa H, Kanda K, Hirose
H. A study to determine necessity of QOL and ADL
evaluation and care of institutionalized senile elderly.
Report of Grant-in-Aid for Scientific Research from
the Ministry of Education, Science and Culture, 1993.
(in Japanese)
18. Maitoko M, Kanda K. An approach to evaluation ofquality of nursing, focusing on medication errors.
Japanese Journal of Nursing 58: 122-125, 1994. (inJapanese)
19. Kanda K. Proportion of high-acuity patients according to hospital characteristics. Journal of Japan
Academy of Nursing Science 15(4): 9-14, 1995. (inJapanese)
20. Kanda K, Watanuki S, Kunioka T, Tama E, Kojima Y.
The development and increased use of machineryand equipment in nursing: Current availability andusage. Japanese Journal of Nursing Administration 6:
742-747, 1996. (in Japanese)
21. Kanda K, Watanuki S, Kunioka T, Tama E, Kojima Y.
The development and increased use of machineryand equipment in nursing: Issues related to future
development and increased usage. Japanese Journalof Nursing Administration 6: 834-841, 1996. (inJapanese)
22. Sakaguchi M, Kanda K, Sakai I, Chen JX, Tamura Y,
Shiga Y, Takeo K: Survey on the care needs of persons with health problems and their families living
in urban areas of Japan. Journal of Shiga Academic
Society of Nursing Science 1: 15-19, 1996.
23. Kanda K, Koike J. Predicting quantity of care needs
and nursing intensity. In “Measuring Care Needs ofPatients in Medical Care Institutions,” Report ofGrant-in-Aid for Health Sciences Research from the
Ministry of Health and Welfare (pp. 29-35), 1996. (inJapanese)
24. Kanda K, Sato R, Nagata A. Effects of intermittent
wakening on sleep pattern of nighttime caregiver:
An EEG study with a single subject. Journal of Japan
Academy of Nursing Science 17(1): 75-81, 1997. (inJapanese)
25. Kojima M, Horiuchi S, Ota K, Oikawa I, Kaharu C,
Nomura M, Toyomasu K, Hatono Y, Kanda K,
Tanaka Y. A study of nursing systems in transition
in developed countries. Bulletin of St. Luke’s College
of Nursing 23: 49-68, 1997. (in Japanese)
26. Kanoya Y, Domon Y, Sato R, Kanda K. The nurses’
contribution to the quality improvement of care
under the health care policy to contain costs: areview. Japanese Journal of Nursing Administration
7: 668-675, 1997. (in Japanese)

320

27. Kanda K. Association of nursing service with lengthof hospital stay of patients. Japanese Journal of
Nursing 61: 1018-1022, 1997. (in Japanese)
28. Sato R, Osaka Y, Nomura I, Fuse S, Kanda K. Nurses’
selection of analgesics and the frequency of theiradministration to the patients in 24 hours after transcatheter arterial embolization, transcatheter arterial
infusion, and transcatheter hepatic angiography.
Japanese Journal of Nursing Arts 44: 1001-1006,
1998. (in Japanese)
29. Watanuki S, Kanda K, Kimura K, Minamisawa H,
Mori Y, Takeo K, Tsukamoto M. Body weight loss
and BMI(Body Mass Index) decrease of hospitalized
elders with senile dementia. Journal of Japan
Academy of Nursing Science 18(2): 51-62, 1998. (inJapanese)
30. Sato R, Kanda K, Nagatomo M, Kusakari J.
Relationships between task performance of visiting
care agencies and physicians’ orders. Hospital
Administration 35: 177-184, 1998. (in Japanese)
31. Matsumoto A, Kanda K. Concept and effects of discharge planning. Japanese Journal of Nursing Arts
44: 710-713, 1998. (in Japanese)
32. Kanda K. Physical Restraint of the Elderly in
Institutional Care Setting and Alternatives to
Restraint Use, Report of Grant-in-Aid for Scientific
Research from the Ministry of Education, Science
and Culture, 1998. (in Japanese)
33. Nagatomo M, Kanda K, Sato R, Kusakari J.
Relationship between physicians’ orders and nurses’
service sphere of activity -- a comparison between
home health care agencies and university hospitals.
Hospital Administration 36: 5-16, 1999. (in Japanese)
34. Kanda K, Sato R, Kodama S, Takemura Y, Matsumoto
A. Relations between nurse-staffing levels andadverse outcomes. In “Economic Evaluation of
Nursing Services,” Report of Grant-in-Aid for Health
Sciences Research from the Ministry of Health andWelfare (pp.16-41), 1999. (in Japanese)
35. Sato R, Kanda K, Anan M. Sleep patterns of middleaged and older female family caregivers providing
routine nighttime care for elderly individuals at
home. Journal of Japan Academy of Nursing Science20(3): 40-49, 2000. (in Japanese)
36. Kanda K, Takemura Y. Research agenda for criticalpathways. Japanese Journal of Nursing Research 33:
221-230, 2000. (in Japanese)
37. Kanda K, Sato R, Matsumoto A, Yamagishi M,
Takemura Y. Nurse Staffing and Nurses’ Job by Typeof Healthcare Facility -- Job and Role Expectations ofNurses in Health Services Facilities for the Elderly,
Report of Grant-in-Aid for Health Sciences Researchfrom the Ministry of Health and Welfare, 2000. (inJapanese)
38. Zhao H, Kanda K. Translation and validation of theStandard Chinese version of the EORTC QLQ-C30.
Quality of Life Research 9: 129-137, 2000.
39. Takemura Y, Kanda K. The elements and the processof ‘good nursing practice’ perceived by nurses.

Japanese Journal of Nursing Research 34: 329-339,
2001. (in Japanese)
40. Kanda K, Sato R, Emori K, Takemura Y, Matsumoto
A, Ogawa R. Nurse Staffing and Nurses’ Job by Typeof Healthcare Facility -- Role and Job of Nurses inGeneral Clinics, Report of Grant-in-Aid for Health
Sciences Research from the Ministry of Health andWelfare, 2001. (in Japanese)
41. Kanda K, Zhao H, Takahashi Y. Quality of life inChinese patients with gynecological cancer.
Proceedings of the 2nd Conference on Nursing
Education & Research between School of Nursing,Seoul National University and School of Health
Sciences & Nursing, The University of Tokyo : 6-10,
2001.
42. Kawana N, Ishimatsu S, Kanda K. Psycho-physiological effects of the sarin attack on the Tokyo subway
system. Military Medicine 166 Suppl. 2:23-26, 2001.
43. Ibe T, Ichiduka K, Kawano R, Kanda K, Moriwaki M,
Yanagisawa H. Actual Condition of Nurses as ClinicalResearch Coordinators: Comparison of the Results
Surveyed in 1998, 1999 and 2000. Japanese Nursing
Association, 2001. (in Japanese)
44. Kanda K, Takemura Y, Ogawa R, Yamagishi M,
Kodama S. Nurse Staffing and Nurses’ Job by Type of
Healthcare Facility -- Role and Job of Nurses in
Sanatorium-type Wards of General Hospitals, Reportof Grant-in-Aid for Health Sciences Research from
the Ministry of Health and Welfare, 2002. (inJapanese)
45. Yamagishi M, Kanda K, Takemura Y. Development ofmethods for detecting occurrences of adverse events
as nursing quality indicators. Journal of Japan Societyfor Health Care Management 3: 363-369, 2002. (inJapanese)
46. Matsumoto A, Kanda K, Shigematsu H. Development
and implementation of a critical pathway for abdominal aortic aneurysms in Japan. Journal of VascularNursing 20:14-21, 2002.
47. Kanda K, Ozawa M, Irimura R. Allocation of nursing
time at university hospitals in Japan. Journal of
Nursing Studies National College of Nursing, Japan 1:
17-26, 2002.
48. Kanda K, Kodama S. Determinants of nursing service
in urban public hospitals: An economic perspective.
Nursing and Health Sciences 4: 63-72, 2002.
49. Takemura Y, Kanda K, Matsumoto A, Yamagishi M.
Role expectations of nurses in health service facilities
for the elderly. Nursing and Health Sciences 4: 73-83,
2002.
50. Sato R, Kanda K, Anan M, Watanuki S. Sleep EEG
patterns and fatigue of middle-aged and older femalefamily caregivers providing routine nighttime carefor elderly persons at home. Perceptual and Motor
Skills (in press).

321

Department of
Adult Nursing / Terminal andLong-term Care Nursing
Outline and Research Objectives
The origin of the Department of Adult Nursing /Terminal and Long-term Care Nursing
was “Department of Adult Health” in the School of Health Sciences (1965-1992), and afterthat “Department of Adult Health and Nursing” in the School of Health Sciences & Nursing(1992-).
From 1995 to 1997, the Graduate School of Medicine shifted to the chair system ofGraduate Schools, and our new two departments were established. The members of two
departments cooperate in education and research together.
Our major research objectives are concerning followings issues.
1. Cancer Nursing
2. Adult Health Nursing
3. Nursing Care System for Outpatients
4. Evaluation of Palliative Care Services
5. Spiritual Care for Terminally Ill Persons and Their Families
6. Continuity in Palliative Care System
7. Dissemination of Palliative Care
8. Methodology of Nursing Research and Education

Faculties and Students
Professor and Chair

Keiko Kazuma, M. of Hlth.Sc., The
Univ. of Tokyo, School of Health
Sciences, 1976; Dr. Hlth.Sc., The
Univ. of Tokyo, 1989. Since 1999
Lecturer
Masako Kawa, M. of Hlth.Sc., The
Univ. of Tokyo, School of Health
Sciences, Since 1996
Associate .................................2
Postdoctoral Fellow ................4
Graduate student ..................14 (Dr. program;6, Ms. program;8)
Research student.....................7
Secretary .................................1

Past Research and Major Accomplishments
1. Cancer Nursing
One of the main subjects of our cancer nursing
research is cancer risk counseling, especially in genetic
counseling of patients and /or kindred of patientswith familial cancer syndrome, expressed in adulthood. In genetic counseling, ethical consideration isvery important for application of gene testing. At
present, prophylactic surgery is the first choice as a322

preventive strategy for familial cancer syndrome
expressed in adulthood. We have studied clients’needs and the role of nurse-counselors, based on theirbackground of familiarity and practical experienceswith perioperative management and intervention foradaptation to the patients’ altered intestinal structure
after surgery. We developed a decision tree for theprocess of cancer risk counseling based on the foreign
literature and our clinical experience with familial
adenomatous polyposis family. The decision tree wasagreed with peer group of cancer genetic counselingand is introduced in seminars for familial cancer counselors.
Another concern is intervention to facilitate adaptation after cancer surgery. We developed a measurement scale for feeling of stability in daily life with astoma for people with a permanent colostomy
because of malignancy, and explored the factors influencing the feeling of stability. The results showedthat the frequency of the stoma clinic and support
from nurses contributed to that feeling
2. Adult Health Nursing
The domain of adult health nursing contains work
concerning health promotion and symptom management. For the former, a brisk walking program wasapplied for the middle nurse managers, a group thattends to lack of exercise. Several positive effects inserum characteristics were approved in the intervention group compared to control group.
The subjects of symptom management researchwere patients who underwent surgery for gastrointestinal disease or inflammatory disease. After gastrectomy, we found that increased symptoms wererelated to changes in eating behavior associated withreturn to the workplace, and it was suggested thenecessity of intervention after discharge. A measurement scale was also developed to evaluate difficulties
in daily life after surgery for oral tumor.
The subjects of symptom management for inflammatory disease were patients with ulcerative colitis(UC) or rheumatoid arthritis (RA). UC patients experience difficulties in daily life related to decreased vigor
and altered defecation. We developed an instrument
to measure the difficulties of life, and explored thefactors influencing it. For RA patients, a series of studies was carried out on association with fatigue, coping
behavior, and nutrition, and an exploratory study was
implemented on home exercise and the factors influencing it.
3. Nursing Care System for Outpatients
Recently, nurses’ role in hospital outpatient departments and in home care settings has been changing
dramatically, because of the shortening of hospital
stays, changes in types of disease, and increasing elderly population. Several surveys were conducted onnurses’ recognition of the care needs of outpatients,
evaluation of nurses’ activities for outpatients, andfact-finding about consultation and teaching activities
by nurses throughout Japan (dental university hospital, and all hospitals above 200 beds).
4. Evaluation of Care Services in Palliative Care
Institutions
It was difficult to investigate quality of life in vulnerable patients, so that it was assumed that theeffectiveness of palliative care could be clarified byclinical audit, which is the systematic critical analysisof the quality of clinical care by the audit tool insteadof the QOL questionnaire. Therefore research by theaudit tool was instituted. The feasibility of the
Support Team Assessment Schedule (STAS) and
Edmonton Symptom Assessment System (ESAS),developed abroad as an audit tool, was examined in
two palliative care units. In this study we collaborated
with a research group investigating the reliability andvalidity of the Japanese version of the STAS.
5. Spiritual Care for Terminally Ill Persons andTheir Families
This theme resulted from the spiritual pain discovered in reviewing patients’ complaints of distressing
conditions recorded on their charts. The study on

spirituality was undertaken as collaborative researchwith other universities. After a literature review, interviews were conducted with patients admitted to palliative care units, and the data from the interviews
were analyzed by Grounded Theory Approach.
6. Continuity in the Palliative Care System
We used a questionnaire to investigate the status
and needs of people attending hospice clinics. The
results showed that consultation in which patientsdecide where to stay should be provided at an appropriate time before referral to a palliative care institution.
7. Promotion of Palliative Care;
A member of the teaching staff of our department
joined the working group on “Attitudes toward terminal care” in research implemented by the Ministry of
Health and Welfare in 1998. The results showed thatpalliative care needed to be further understood byboth the general public and the clinical staff. We subsequently conducted a survey of nurses working in
general hospitals to investigate the difficulties
encountered in the care of terminally ill cancer
patients and related factors.
8. Methodology of nursing research and education;
This contains several issues, including surveys andcritiques for application of statistical techniques innursing research, review of utilization of the concentric circle model of body extremities as a non-invasive
measurement tool, and concerning the nursing
process, nursing diagnosis, and terminology in nursing.

Current Research
1. Cancer Nursing;To establish nurses’ role in cancer genetic counseling in Japan, we have joined a research project with
MDs in a continuous study group.
Negotiations concerning to a prospective cohort
study of people with a stoma is in progress with theaim of collaboration between our group and otherinstitutions concerning the process of how to acquirea feeling of stability
2. Adult Health Nursing
Concerning about health promotion, we started acollaborative research program on physical activity
and physical component in frail elderly with a Koreangroup at Seoul National University to promote health.
Symptom management research is conducted in
UC patients after total colectomy. Difficulties in a tem-323

porally ileostomy period are focused on. We are analyzing the cross-sectional data from 84 patients
whether related or not to body weight loss in that
period. And despite of case studies with a few
patients, we ascertained that body weight loss andhypoglycemia were not appeared in patients forwhom educational intervention was period about how
to eat in that period.
3. Nursing Care System for Outpatient
Nursing function and efficiency in hospital outpatient department is being surveyed with diabetics notusing insulin as the model, and a prospective cohort
study is being planned.
4. Evaluation of Palliative Care Services
The study of the reliability and validity of theJapanese version of the STAS has just been completed. We are now preparing a booklet to encourage useof this tool in clinical settings. We are also participating in a study group that is establishing evaluation criteria according to the peer review method developedin the UK.
5. Spiritual Care for Terminally Ill Persons andTheir Families
Interview data are being analyzed and some of theresults are being prepared for submission to a foreign
journal. Next, we must plan a study on appropriate
spiritual care for terminally ill persons.
6. Continuity in the Palliative Care System
We obtained a grant for Day Care research.
Preparation for needs assessment prior to development of a day care program is in progress. Researchon consultation and support for patients discharged
from cancer centers is also continuing.
7. Promotion of Palliative Care;
Barriers to palliative care are being examined topromote palliative care in general practice. In addition, research on palliative care for other chronic illnesses is being planned.
8. Methodology of nursing research and education
Projects of terminology in nursing are being continued in International Classification of Nursing
Practice (ICNP) and nursing actions in Japan.

Future Projects
Both departments intend to continue to develop
current research projects. A new project is being considered: making the measurement scales we developed available on our homepage. This project will be

324very useful in refining the scales and compiling clinical data.

Research Grants
1. Keiko Kazuma: The Ministry of Education, Culture,
Sports, Science and Technology (MEXT), Grant-inaid for Scientific Research (C); The Development of
Nursing strategy for the postoperative patients withlower gastro-intestinal Diseases (2,300,000 yen,
Representative researcher, 2000-2003).
2. Keiko Kazuma: The Ministry of Health, Labor andWelfare (MHLW), Grant-in-aid for Scientific
Research; Nursing function of outpatient department
in hospital for chronically ill people (6,500,000
yen;2002, Collaborative researcher, 2002-2003).
3. Keiko Kazuma: Japan Society for the Promotion of
Science (JSPS), The Japan-Korea Basic Scientific
Cooperation Program; A Comparison of Bio-psychosocial status related to Physical Activity of theElderly between Korea and Japan: Biophysical Index,
Health status, Competence of Daily Activity andDepression. (2,400,000 yen, Representative
researcher, 2002-2004).
4. Masako Kawa : The Mitsubishi Foundation, Grant
for Social Welfare Activities and Research; Basic
study on the development of spiritual care for thewell-being of terminally ill patients with cancer.
(2,000,000 yen, 2000-2001.)
5. Masako Kawa : Japan Society for the Promotion of
Science (JSPS), Grant-in-aid for Scientific Research(C); The development of palliative day care program
for the persons with advanced cancer. (3,900,000
yen, Representative researcher, 2002-2004.)

Select Publications
1. Imamura Y, Ozawa T, Miyashita M, Kawa M, Kojima
M. Consultation service at hospice clinic and information given at referrals on hospice and palliative
care: The attitude among terminally ill patient withcancer and family members. J Jpn Soc Cancer Nurs
13(2): 60-68., 1999. (in Japanese)
2. Ojima T, Nakamura Y, Hashimoto S, Miyashita M,
Hayashi M, Kato M, Matsumura Y, Fukutomi K. The
development of the regional health and welfare
indexes. Journal of Health and Welfare Statistics
46(15): 3-9., 1999.(in Japanese)
3. Takeda Y. Parents’ attitudes to predictive genetic
testing in their children for familial adenomatous
polyposis. The Ochanomizu Medical Journal 47(3):129-146., 1999. (in Japanese)
4. Koike T. The relationship between fatigue and nutritional status in patients with rheumatoid arthritis.
The Ochanomizu Medical Journal 48(3･4): 95-107.,
2000. (in Japanese)
5. Furuse K, Kawahara M, Nishiwaki, Fukuoka M,
Takada M, Miyashita M, Ohashi Y. Phase I/II studyof vinorelbine, mitomycin, and cisplatin for stage

IIIB or IV non-small-cell lung cancer. Journal of
Clinical Oncology 17(10): 3195-3200., 1999.
6. Ito N, Miyashita M, Kojima M. The self-care ability
and medical/social support in ostomates. J Jpn Soc
Cancer Nurs 1999. 13(1): 25-34., (in Japanese)
7. Miyashita M, Hashimoto S, Kawa M, Kojima M.
Attitudes towards terminal care among the general
population and medical practitioners in Japan.
Japanese Journal of Public Health 46(5): 391-400.,
1999. (in Japanese)
8. Matsumura Y, Nakamura Y, Hayashi M, Kato M,
Miyashita M, Ojima T, Hashimoto S, Fukutomi K.
Prefectural differences of the proportion of the smoking population. Journal of Health and Welfare
Statistics 46(6): 23-28., 1999. (in Japanese)
9. Miyashita M, Hashimoto S, Ojima T, Nakamura Y,
Hayashi M, Kato M, Fukutomi K. An estimation of
the proportion of handicapped people and healthexpectancy for aged person: prefectural estimation
based on existing health statistical resources. Journalof Health and Welfare Statistics 46(5): 25-29., 1999.
(in Japanese)
10. Hashimoto S, Miyashita M, Tsuji I, A comparison of
calculation method for health expectancy. Journal of
Health and Welfare Statistics 46(4): 12-16., 1999. (inJapanese)
11. Hayashi M, Hashimoto S, Kato M, Ojima T,
Nakamura Y, Miyashita M, Fukutomi K. The importance or regional health and welfare indexes. Bulletinof Fukushima school of nursing 1: 33-38., 1999. (inJapanese)
12. Kazuma K, Kawagoe H, Kawamura S, Yamazaki M.
Development of the nursing protocol for management of medical treatment in home care, (1) The fact
finding study of visiting nurse station providing
management of medical treatment. Kango 51: 110116., 1999. (in Japanese)
13. Kazuma K, Kawagoe H, Kawamura S, Yamazaki M.
Development of the nursing protocol for management of medical treatment in home care, (2) The fact
finding study of visiting nurse station providing
management of medical treatment. Kango 52: 108114., 1999. (in Japanese)
14. Kitano K, Kawamura S, Kazuma K. A study on working women caring for their husbands with an
intractable disease. J J Intrac Ill Nurs 3: 53-66., 1999.
(in Japanese)
15. Mizuno Y, Ogura A, Kawamura S, Kazuma K. Studyon planning and nursing management for safe outside activities for ventilator-assisted-individuals. J J
intrac Ill Nurs, 3: 42-52., 1999. (in Japanese)
16. Amano S, Kawamura S, Kazuma K, Ushikubo M.
Protocol of pressure ulcers management in homehealth care-algorithm of nutritional assessment andmanagement. J Jpn Acad Home Health Care, 2: 5159., 1999. (in Japanese)
17. Kazuma K. Utilization of the concentric circle modelin clinical nursing: a Review. Journal of Medical and
Dental Sciences. 46(4): 139-143., 1999.

18. Takeda Y, Kazuma K. The nurses’ role in cancer
genetic testing and counseling. Nursing Today, 14:
72-74., 1999. (in Japanese)
19. Kazuma K. The Current of nursing process and nursing diagnoses. Jpn Nurs Admin, 9: 671-677., 1999.
(in Japanese)
20. Kawa M. Supporting the bereaved family. In Ohba
M, Endo E, and Inayoshi M (eds.). The new cancer
nursing, Brain Press, Tokyo 361-365., 1999. (inJapanese)
21. Kawa M. The subjects to be studied in palliative care.
In Ohba M, Endo E, and Inayoshi M (eds.). The newcancer nursing, Brain Press, Tokyo 366-374., 1999.
(in Japanese)
22. Kazuma K, Takeda Y: Genetic counseling in cancer
nursing, in Ohba M, Endo E, Inayoshi M (eds.). New
cancer nursing, Brain Press,: 59-71., 1999. (inJapanese)
23. Okusaka k, Kazuma K. Investigation of changes insymptoms and eating behavior associated with thereturn to the workplace of postgastrectomy patients.
J Jpn Acad Nurs Sci 20(3): 60-68., 2000.
24. Koike T, Kazuma K, Kawamura S. The relationshipbetween fatigue, coping behavior, and inflammationin patients with rheumatoid arthritis. The Japan
Rheumatism Association and Springer-Verlingere
Tokyo 2000. Mod Rheumatol. 141-149, 2000.
25. Hashimoto M, Koike T. Development of instrument
to measure difficulties of life in patients with ulcerative colitis. The Ochanomizu Medical Journal 48(2):71-76., 2000 (in Japanese)
26. Aoki H, Sakai E. Proposal of a measurement scale toevaluate difficulties of daily life in patients after surgery of oraltumor. The Ochanomizu Medical Journal
48(2): 63-70., 2000.(in Japanese)
27. Koyama C, Kawa M, Kasahara Y. Progression of disease and process of psychological change of terminally ill patients with cancer, An analysis of medical
and nursing charts. Japanese Journal of ClinicalThanatology 5(2): 85-91., 2000. (in Japanese)
28. Koike T, Kazuma K. Recognition by nurses of theneed for outpatient consultation and guidance, andchallenge for the future. J Jpan Acad Nurs Admin
Policies 4(1): 23-32., 2000. (in Japanese)
29. Kazuma K, Yokomura T, Hara M, Aoki H, Okamoto
N. Practice and evaluation of primary nursing in outpatient department. J Jpan Acad Nurs Admin Policies
4(1): 34-38., 2000. (in Japanese)
30. Kato M, Hashimoto S, Miyashita M, Hayashi M,
Nakamura Y, Ojima T, Matsumura Y, Fukutomi K.
The transition of elderly welfare policy differences in
prefectures and municipalities. Journal of Health andWelfare Statistics 47(4): 8-13., 2000. (in Japanese)
31. Ushikubo M, Kazuma K. Research and challenge forthe care of chronically ill people. J J Nurs Res 33: 203211., 2000. (in Japanese)
32. Takeda Y, Kazuma K. Family cancer and nursing.
Gastroenterolog Surg Nurs 15: 205-211., 2000. (inJapanese)32533. Sato M. How to assessment defecation dysfunction
and how to approach these patients. The Japanese
Journal of Nursing Art 46(11): 26-32., 2000. (inJapanese)
34. Kawa M. The nursing role in palliative care. In:
Higashiha M and Kondo M (eds.). Palliative Care.
Nursing QOL Books. Igakushoin. Tokyo 117-128.,
2000. (in Japanese)
35. Kazuma K. Risk counseling for familial cancer. Journalof Family Tumor 1(1): 9-15., 2001. (in Japanese)
36. Hayakawa E, Ushikubo M, Kawamura S, Kazuma K,
Yuasa S, Takagi K. Analysis of the activity of the
medical counseling services for intractable illnesses(Nanbyo) of the Tokyo medical association. Jpn J
Prim Care 24(3): 189-196., 2001. (in Japanese)
37. Okano Y, Okamura H, Watanabe T, Narabayashi M,
Katsumata N, Ando M, Adachi I, Kazuma K, Akechi
T, Uchitomi Y. Mental adjustment to first recurrence
and correlated factors in patients with breast cancer.
Breast Cancer Res Treat 67(3): 255-262., 2001.
38. Murakami Y, Gondo N, Okamura H, Akechi T,
Uchitomi Y. Guilt from negative genetic test findings.
Am J Psychiatry 158(11): 1929., 2001.
39. Kazuma K, The relationship between fatigue andnutritional status in patients with Reumatoid
Arthritis. The international conference. Tokyo
University, Oita University & S.N.U. Seoul: 109-112.,
2001.
40. Agata T, Kawa M. Research on medicine and nursing. In Agata T. (ed.), PubMed, Impact Factor for
EBM. Chugaiigakusha, Tokyo:1-20., 2001. (inJapanese)
41. Takeda Y, Kazuma K. Practice of genetic nursing; cancer. In Andou H. et al (eds.), Genetic Nursing.
Ishiyaku Publishing Co., 108-118., 2001. (in Japanese)
42. Imamura Y. Hospice home care. Kango 53(10): 98102., 2001. (in Japanese)
43. Takeda Y. Need of the support system for family
members with hereditary cancer. The Japanese
Journal of Nursing 65(10): 948–951., 2001. (inJapanese)
44. Aoki H, Ohta K, Kazuma K. The current status andproblems of nursing for outpatients department oforal surgery of dental university hospitals in Japan.
The Journal of the Stomatological Society, Japan
69(2): 9-14., 2002. (in Japanese)
45. Emoto A. A study on the long-term effects and contributing factors of pelvic floor muscle exercise in
women with stress urinary incontinence. The
Ochanomizu Medical Journal 50(1): 19-34., 2002.(inJapanese)
46. Watanuki E. Nursing staff’s understanding of their
liability in nursing practice. Jpn Nurs Res 25(2): 6169., 2002. (in Japanese)
47. Sato N, Kato A, Sugimura H. Physician’s level ofknowledge regarding genetic counseling in the fieldof clinical genetics. Journal of Familial Tumor 2(1):20-25., 2002. (in Japanese)
48. Imamura Y, Kawa M, Kayama M, Mizuno M, Otsuka
M, Murata H. Conceptual structure on spirituality in

326

end-of-life patients with cancer. The Japanese
Journal of Hospice and Palliative Care 12(5): 425434., 2002. (in Japanese)
49. Kawa M, Koyama C, Imamura Y, Miyashita M,
Ashiya T, Kasahara Y. Physical state and psychological expression of terminally ill patients with cancer
in a palliative care unit. The Japanese Journal of
Clinical Thanatology 7(1)., 2002. (in press)
50. Furukawa F, Kazuma K, Kawa M, Miyashita M, Niiro
K, Kusukawa T, Kojima M. Effects of an off-site
walking program on energy expenditure, serum
lipids, and glucose metabolism in middle-aged
women. Biological Research for Nursing 4(3)., 2002.
(in press)Department of
Family Nursing
Outline and Research Objectives
The Department of Family Nursing was established in April 1992 and was started at
October 1992 when Dr.Sugishita had a position of Professor of Family Nursing. Currently, it
has four faculty members; one professor, one associate professor and two associates.
Our department is based on new concept which combines together the following four
nursing fields: maternal-child nursing, pediatric nursing, gerontological nursing and family
nursing.
Our department provides lectures, clinical practicums and fieldwork on the four nursing
fields to the undergraduate students of School of Health Sciences and Nursing. Our department also gives two required lectures, that is, “clinical immunology ” and “laboratory methods in health science” to the undergraduate students of our school.
We also have responsibilities to give special lectures, advanced seminar and laboratory
and/or field work on family nursing to the postgraduate (master’s and doctoral) students.
Our research field concerns the development of methodology to improve nursing skills
and techniques, and the development of physical, psychological and interactive assessment
procedures for patients and families in our field.

Faculties and Students
Professor and Chair

Chieko Sugishita R.N., Ph.D.
1992~
Associate Professors Kiyoko Kamibeppu R.N., Ph.D.
Associate .................................2
Postdoctoral Fellow ................5
Graduate student ..................10
Research student...................15
Secretary .................................2

Past Research and Major Accomplishments
Topics of our past research projects were as follows.
①Control of maternal-child transmission of microorganism, ②Control of body fluid circulation in
women supported with physical compression, ③
Evaluation of antiboby development after immunization in children and analysis of immune system, ④Environment/food factors of allergic diseases amongchildren, ⑤Recent anxiety coping strategies in Japan,⑥Psychological factors of the family caregiving forthe frail elderly, ⑦Development of new devices forevaluating autonomic nervous activities of the frail
elderly, ⑧Development of Japanese version of family
functioning survey.

Major accomplishments were as follows.
Number indicates the number of research project.
①We examined the samples from neonates, theirmothers, medical staffs and environment in amaternal ward to isolate some bacteria in order todemonstrate the route of transmission toneonates. Some of the neonates had the same bacteria as their mothers, medical staffs and environment at the point of discharge. Therefore, Several
bacteria could have been transmitted to theneonates from their mothers, medical staffs
and/or environment in hospital.
Our data also showed that mamma trouble did
not decrease even if they use nipple wiped withsterilized cotton. It seemed that hand washing
before breastfeeding was more important.
② The physical compression with stocking was effective to control the blood pressure decrease whensupine position was changed to standing up position, especially in the elderly. It was suggested thestocking might prevent the orthostatic hypotension in the elderly.
③Vaccine-induced immunity of measles, mumps,
rubella and chickenpox at one to three years ofage continued until nine years of age. But the levelof immunity was lower than naturally aquired
immunity.

327

④This study showed that incidence of atopic children
was decreased by age, but any risk factors could
not be found in atopic children compared with
non-atopic children.
⑧We developed Japanese version of Feetham Family
Functioning Survey (FFFS)which enables to evaluate the family functions for young families withchildren.

Current Research
Current research projects include four fields, thatis, maternal-child nursing, pediatric nursing, gerontological nursing and family nursing.
Topics of our current research projects are as follows: ①Control of hospital infection such as transmission of bacteria including MRSA between mothers
and their babies, ②Exploring the new nursing caremodel for the children with disabled and/or chronic
illness in the community, ③Interaction between nursing home residents and their families, ④Nurses’ interests in family care for critical patients, ⑤Assessmentof autonomic nervous activity in the elderly withchronic illness in hospital, ⑥Incidence of adverse
reactions associated with Japanese-style medical
acupuncture for the elderly, ⑦Follow up study ofphysical, mental and social activities of patients withdementia.

Future Prospects
Since two more Departments, ‘Midwifery Nursing
and Women’s Health’ and ‘Gerontological Nursing’,
will be established in 2002 and 2003 respectively, ourdepartment will be responsible mainly for Family
Nursing and Pediatric Nursing.
Concept of family nursing was introduced to Japancountry only ten years ago, and The Japanese
Association for Research in Family Nursing was
founded by our department in 1994. Our department
will explore the establishment of the theory in family
nursing collaborating with its research, practice andeducation.
Education and research of pediatric nursing arealso involved in our activities. We intend to research
new fields of pediatric nursing in the viewpoints ofchild rights, good prognoses of severe illness, birth ofchildren with diversified origins on the basis of the
progress of reproduction technology, and well-being
for all of the families.

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Research Grants (within 5 years)
1, Chieko Sugishita, Naohiro Hohashi: Infection control among nursing mothers and neonates, andrelated quality of nursing care -According to analyses of bacteria including MRSA in a maternity clinic-, Grants-in-Aid for Scientific Research (B)(2),
1998-1999, The Ministry of Education, Culture,
Sport and Science, ¥10,200,000
2, Chieko Sugishita, Noriko Yamamoto, Mamoru
Kumada: Development of passive of leg cycle exercise for the elderly to improve the walking rhythm
and enhance the autonomic nervous activity.
Grants-in-Aid for Scientific Research (B)(2), 19992000, The Ministry of Education, Culture, Sportand Science, ¥6,400,000
3, Chieko Sugishita, Kunihiko Hayashi, Naohiro
Hohashi, Kunie Mituhashi: Development of two
dimensional scale in order to assess the familysystem and care ability, Grants-in-Aid for
Exploratory Research, 1999-2000, The Ministry of
Education, Culture, Sport and Science, ¥2,100,000
4, Hiroko Torii, Hideko Mori, Chieko Sugishita:
Study on nurses’ views on family and family nursing, Grants-in-Aid for Exploratory Research, 19992000, The Ministry of Education, Culture, Sportand Science, ¥800,000
5, Chieko Sugishita, Kiyoko Kamibeppu, Keiko
Murata, Naohiro Hohashi, Hiroko Torii, Makiko
Owaki, Kunihiko Hayashi: Development of nursing practice model for the children with chronic
illness or/and disabilities and their families incommunity. Grants-in-Aid for Scientific Research(B)(1), 2001-2003, The Japan Society for the
Promotion of Science, ¥12,100,000

Select Publications
1. Sugishita C, O’shea S, Best J, and Banatvala J E.
Rubella serology by solid-phase radioimmunoassay.
its potential for screening programmes.
Cline.Exp.Immunol 31(1), 50-54, 1978.
2. Shishdo A, Sugishita C, Takahashi C, Sakata H,
Hirayama M, and. Kimura M. A ten-year follow-up
study on measles vaccination in Japan: Evaluation of
the efficacy analyzed on a computer system.
Japanese Journal of Medical Science & Biology 31(4),339-356, 1978.
3. Kono R, M. Hirayama, Sugishita C, and Miyamura K.
Epidemiology of rubella and congenital rubella infection in Japan. Reviews of Infectious Diseases,
Supplement 1, 56-63, 1985.
4. Maekawa K, Nozaki H, Fukushima K, Sugishita C,
and Kuriya N. Clinical analysis of measles, mumps
and rubella vaccine meningitis?Comparative study ofmumps, mumps meningitis and MMR meningitis?.
Jikeikai Medical Journal 38(4), 361-368, 1991.
5. Iida Y, Sugishita C, Matsuura K, and Torii H. English
education in colleges of allied health sciences.
Bulletin of Tokyo Metropolitan College of Allied
Medical Sciences 7 97-102, 1994.
6. Nagase H and C. Sugishita. Mother’s anxiety related
to their child rearing on telephone consultation.
Analysis of statement about the in health center-. J
Child Health 53, 668-76, 1994. (in Japanese)
7. Sugishita C, Tezuka K, Hohashi N and Higurashi M.
Effects of medical compression stockings on lower
leg girth in healthy women. Jpn J Maternal Health
35,50-53, 1994.(in Japanese)
8. Hohashi N, Etoh T, Kurahashi T, Tejima R and
Sugishita C. Seroepidemiology of primary school children in the third grade for naturally acquired or vaccine
inducedimmunityofmeasles,mumps,rubella,and chickenpox determinedby presence of IgG antibody. Infection and Immunity
in Childhood 6,251-4, 1994. (in Japanese)
9. Sugishita C, Mori H, Nagase H, Tezuka K, Kurahashi
T and Hirayama M. Immunization of communicabledisease and morbidity rate in handicapped children
and young adults. J Child Health 53, 842-8, 1994. (inJapanese)
10. Tamura T, Naoko H, Nagase H, Morokata T, Igarashi
O, Oshimi Y, Miyazaki S and Nariuchi H. Early activation signal transduction pathways of Th1 and Th2cell clones stimulated with anti-CD3. J. Immunol 24,
4692-4701,1994.
11. Yoshimoto T, Nagase H, Nakano H, Matsuzawa A
and Nariuchi H. AVβ8.2-specific superantigen fromexogenous mouse mammary tumor virus carried by
FM mice. Eur.J.Immunol. 24, 1612-1619, 1994.
12. Sugishita C, H Yonezawa, N Hohashi, Y Iida, F
Komoda, T Iwata and H. Hayakawa. The significanceof guidance for mothers of allergic children withregard to dietary restrictions. Proceeding of The

XVth World Conference of the International Union
for Health Promotion and Education. 312-315, 1995.
13. Sugishita C, Morotomi C, and Murata I, etc.
Evaluation of PHA/RIA antibody responses to CHO
cell derived HB vaccine compared with yeast andplasma derived HB vaccines. Clinical Virology 23,
231-236, 1995.
14. Sato Y, Sugishita C, Kataoka Y. The effect of walking
on blood pressure changes and accelerated plethysmogram (APG) in middle aged and elderly people
with hypertension. Jpn J Res Fm N 2, 28-35, 1996.
15. Hohashi N, Hayashi T, Fusaki N, Takeuchi M,
Higurashi M, et al. The protein tyrosine kinase Fyn
activates transcription from the HIV promoter via
activation of NF K b-like DNA-binding proteins. Int.
Immunol 7, 1851-1859, 1995.
16. Tezuka K, Sato R, Igarashi T and Sugishita C. The
effect of pneumatic massage on the leg circumferences of healthy women. Clinical Report 30, 203-209,
1996.
17. Tezuka K, Sugishita C, Kuwaki T, Higurashi M and
Kumada M. Effects of compression stockings onblood pressure and its orthostatic change in female
subjects. Pathophysiology 4, 81-86, 1997.
18. Iwata T, Hayakawa H, Komoda F, Koseki M,
Miyakawa T, Sakurayama T, Yamasaki K, Honjou M,
Hirakawa N, Inoue K, Hohashi N, and Sugishita C.
Prospective study of atopic status in infants of thecohort in Tokyo,Japan. Allergology International 46,
221-225, 1997.
19. Miki A, Haratani T, Sugishita C, Kawakami N, Araki
S, and Kawamura N. The effects of job stressors on
depression and job satisfaction among hospital nurses in Japan. Proceedings 15th Asia Conference On
Occupational Health 226-231, 1997.
20. Yoshidome A, C. Sugishita.A study on the relationship between mamma trouble and practice of nipple
wipping with sterilized cotton. Japanese Journal of
Maternal Health 38, 350-354, 1997. (in Japanese)
21. Tsuchiya H, Kusaka S, Hohashi N, Sugishita C. The
decrease of the seroprevalence of antibodies against
herpes simplex virus and cytomegarovirus among
students of nursing college and consideration of
their lifestyles and rearing environments. Infection
and Immunity in Childhood 9, 95-102, 1997. (inJapanese)
22. Page J, Lim H, Lynett L-Y, Heller R, Fukuhara H, Ban
S, Yamamoto N, and Kurokawa K. Critical appraisal
workshop in Japan: a short term evaluation.
Education for Health. 10, 371-377, 1997.
23. Yamamoto N and Wallhagen M I. The continuationof family caregiving in Japan, Journal of Health andSocial Behavior. 38, 164-175, 1997.
24. Nagase H, Wang C, Yoshimoto T, Sugishita C, andShiroishi T, Matuzawa A and Nariuchi H. Analysis ofnovel mutant mice secreting soluble CD4 without
expression of membranebound CD4. Eur.J.Immunol
28, 403-412, 1998.

329

25. Naito N, Hashimoto Y, Sugishita C. Development of
the childcaring perspective scale for mothers withchildren aged three or younger(CPS-M 97): -Validity
and Reliability of the CPS M 97-. Journal of Japan
Academy of Nursing Science 18, 1-9, 1998. (inJapanese)
26. Torii H and Sugishita C. Behavioral problems of thechild when his/her mother is stating with his/her
hospitalized sibling. Japanese Journal of Research in
Family Nursing 4, 18-23, 1998. (in Japanese)
27. Matsui N, Ju K, Kumada M and Sugishita C. The
effect of water exercise with a jet on the frail elderly.
The Autonomic Nervous System 35, 419-423, 1998.
(in Japanese)
28. Yamamoto N and Sugishita C. Utilization patters ofcare assistance by family caregivers of visitors to aclinic for the elderly. Japanese Journal of Gerontology
19, 129-139, 1998.
29. Yoshinaga, A., and Horibe, H. Serum lipid concentrations in Japanese children: A synthesis of 27 studies.
CVD Prevention. 1: 55-70, 1998.
30. Yamamoto N, and Wallhagen M I. Service use by
family caregivers in Japan. Social Science andMedicine, 47, 677-691, 1998.
31. Sugishita C. Development of family nursing in Japanpersent and future perspectives. Journal of Family
Nursing 5(2), 239-244, 1999.
32. Matsui N, Ju K, Sugishita C and Kumada M.
Assessment of baroreceptor reflex sensitivity duringpassive leg cycle exercise using bivariate autoregressive model. Proceedings of the IFMBE/IMIA 3rd
International Workshop on Biosignal Interpretation.
91-94, 1999.
33. Omine F, Hamamoto I, Odo S, Miyagi M, Sunagawa
Y, Sugishita C. A Study of the Effect of Peer
Education on Student Sexual Awareness. Journal ofJapan Academy of Nursing Science 19(3), 64-73,
1999. (in Japanese)
34. Nakatani Y, Hohashi N and Sugishita C. Changes ofhemodynamics due to white noise stress according
to the extents of obesity. The Japanese Journal of
Health and Human Ecology 65, 208-218, 1999. (inJapanese)
35. Hohashi N, Matsumoto K and Sugishita C. National
survey of actual conditions related to MRSA conducted through home-visit nursing care station-Focusing
on incidence of MRSA and problems related to homecare-. Japanese Journal of Research in Family Nursing
4, 119-124, 1999. (in Japanese)
36. Mori N, Ito T, Tsushima R, Hhashi N, Hiramatu K
and, Sugishita C. Bacteriological and molecular epidemiological analysis of methicillin-resistant staphylococcus aureus isolated in a maternity hospital.
Environmental Infections 14, 296-302, 1999. (inJapanese)
37. Yamamoto-Mitani N, and Sugishita C. Discharge education for neurology patients and their family:
Comparison between the recognition of nurses,

330

patients, and their family. Japanese Journal of
Research in Family Nursing 5(2), 4-18, 1999. (inJapanese)
38. Yamamoto N M, Tamura M, Deguchi Y, Ito K and
Sugishita C. The attitude of Japanese family caregivers toward the elderly with dementia.
International Journal of Nursing Sutudies 37, 415−
422, 2000.
39. Mori N, Hitomi S, Okuzumi K, Yoneyama A,
Sugishita C, and Kimura S; Susceptibility to vancomycin of methicillin-resistant staphylococcus
aureus isolated in a university hospital in Japan. J. J.
A. Inf. D. 74(11), 966-972, 2000.
40. Yamamoto N M, Abe T, Yamada Y, Yamazato C,
Amemiya H, Sugishita C and Kamata K. Reliability
and validity of Japanese quality of life scale for theelderly with dementia. Nursing and Health Sciences2(2), 69-78, 2000.
41. Matsui N, Ju K, Ohmori N, Sugishita C and Kumada
M. A new device for evaluating autonomic nervous
activity of cardiovascular system. Biomedical
Sciences Instrumentation 36, 87-92, 2000.
42. Maeda M, Hohashi N, Sugishita C. Survey of thefamily accompaniment of hospitalized children.
Japanese Journal Research in Family Nursing 5, 94100, 2000. (in Japanese)
43. Kudo Y and Sugishita C. A Study of infection control
among HPN patients. Infection Control 179-182,
2000. (in Japanese)
44. Yamamoto-Mitani N and C. Sugishita. The effectiveness of nurses’ discharge education and post-discharge patient problem evaluation. Japanese Journalof Nursing Science 20(2), 21-28, 2000. (in Japanese)
45. Hohashi N, Maeda M and Sugishita C. Developmentof the Japanese-language Feetham Family
Functioning Survey(FFFS) and evaluation of its effectiveness. Japanese Journal of Research in Family
Nursing 6, 2-10, 2000. (in Japanese)
46. Yamashita H, Tsukayama H, Hori N, Kimura T and
Tanno Y. Incidence of adverse reactions associatedwith acupuncture. J Altern Complement Med. 6(4),345-350, 2000.
47. Kobayashi N. Formal service utilization of the frail
elderly at home during the last six months of life,
Nursing and Health Sciences 2, 201-204, 2000.
48. H Yamashita, H Tsukayama, and Sugishita C; Local
adverse reactions commonly seen in Japanese-style
medical acupuncture practice. Clinical Acupuncture
and Oriental Medicine 2, 132-137, 2001.
49. Yamashita H, Tsukayama H, White A R, Tanno Y,
Sugishita C, and Ernst E. Systematic review ofadverse events following acupuncture: the Japanese
literature. Complementary Therapies in Medicine, 9,98-104, 2001.
50. Yamamoto-Mitani N, Sugishita C, Ishigaki K,
Hasegawa K, MaekawaN, Kuniyoshi M, and Hayashi
K. Development of instruments to measure appraisalof care among Japanese family caregivers of the eld-

erly, Scholarly Inquiry for Nursing Practice: An
International Journal, 15, 113-135, 2001.

331

Department ofCommunity Health Nursing
Outline and Research Objectives
Department of Community Health Nursing (CHN) was established in June 1992 with theappointment of Dr. Katsuko Kanagawa as the first professor of the department. In May
1993, Dr. Sachiyo Murashima joined as associate professor and took charge as professor
after Dr. Kanagawa’s retirement.
During ten years since the department’s inauguration, 12 students have succeeded ingaining the degree of Doctor of Health Sciences; 19 the degree of Master of Health Sciences,and in the undergraduate course 20 students elected to write theses in community healthnursing.
In order to enhance the practice of nursing and public health, the department strives toundertake research for improvement of quality of life (QOL) of people in various community
settings by focusing on:
1. Developing a comprehensive health care system for community care through publichealth nursing leadership;
2. Intervention projects to demonstrate efficacy of new initiatives directed at improvementof QOL of bedridden, frail and demented elderly;
3. Preventive programs to improve clinical practice in home (e.g. falls) and to reform policy
directions;
4. Undertaking commissioned research with government and non-governmental organizations driven by needs and resources of individuals, families, targeted population groups
and organizations in order to measure cost-effectiveness and quality of care;
5. Developing models for community nursing practice to provide efficient care in partnership with other professionals, families and communities.
The major studies undertaken by the members of the CHN department can be classified
as following;
Directed at

Descriptive studies

Intervention studies

Individuals• Influencing factors of social rehabilitation among former
patients with Hansen’s disease• Meaning of health among cancer patients• Home visiting for schizophrenic patients• Prevention for deterioration of ADL among bedridden elders• Effectiveness of reminiscence for the demented elderly(RCT)• Effectiveness of early discharge planning(RCT)Families• Attitude and interaction of grand children to the demented elders at home • Effectiveness of improved home visit program for• Care of families of children with chronic disease
newborne babies

Aggregates• Characteristics of professional support to family groups of
psychiatric patients• Meaning and strategy of rural women for maintaining both
agriculture and care-giving• Effectiveness of improving support program tofamily groups of psychiatric patients (RCT)Home health service • Community diagnosis and networking• Development and evaluation of around-the-clock
and a community• Comparison between two types of around-the-clock in-home care systems : in-home care system
workinga single type of home help service and a combination type with nursing

Faculty and Students
Sachiyo Murashima, D.Hlth.Sc.,
R.N., P.H.N. (2001~)
Associate Professor Noriko Nishikido, D.Hlth.Sc.,
R.N., P.H.N.
Associates ...............................2
Graduate Students ................11 (7 in Master course; 4 inDoctoral course)
Research Students...................2
Secretaries ...............................3

Past Research and Major Accomplishments

Professor and Chair

332

Research Areas of Community Health Nursing
Community health nursing research has been considered from three perspectives; ‘nursing in the community’, ‘nursing of the community’ and ‘nursing into
the community.’ A majority of studies have focusedon continuity of care and empowering people bystrengthening their potentiality through creating caresystems.

1. ‘Nursing in the community’‘Nursing in the community’ means offering nursing services or health promoting activities to people inthe community in collaboration with other professionals and/or lay people. It is operationalized bydeveloping and testing a new programs to meet needsof individuals or aggregates.
In 1993, when I was transferred to the Universityof Tokyo, care burden for the elderly was a topicalissue in Japan. Following favorable reports of nighttime home helpers’ visiting service, a need to add visiting nurse service was strongly solicited. A study
testing the efficacy of model programs involving four
visiting nurse stations providing around-the-clocknursing care (ACC) commenced with assistance from
the Ministry of Health and Welfare in 1994.
Based on the needs assessment for ACC, an intervention of visiting nursing care early in the morningand evening was provided; and simultaneous evaluation was carried out before, during and after 3
months. Major findings indicated positive effects
related to stabilized medical status, reduced stress,
thereby reduced complications such as decubitus (seearticle number 22).
The results also impacted on policy change: in1996 the fees for ACC visiting nurses, which enabled
the night time visits by nurses, were included in thenational health insurance program. The combination
type of nursing and home help service functioned
more effectively than a single type of home help service (see article number 34).
2. Nursing of the community
Research for ‘nursing of the community’ means
creating care systems in the community for providing
effective care of individuals to enhance their capacity.
A study depicting the role of public health nurses inenabling the community to establish its own care system to solve problems (see article number 25) was in arural town M with a focus on ACC system.
The major purpose of this study was to test thefeasibility of ACC visiting nurse service as desired by
the community, to measure its effectiveness and eventually to develop a policy for providing this service
within the local government health plan. The intervention included establishing the need and acceptance of service, involving residents and other healthworkers, resulting in whole community handling conflicting concepts such as ‘Sekentei’, a kind of socio-cultural norm (see articles number 31, 43, and 48).
Intervention and control population were used forcomparison to measure the effectiveness of aroundthe-clock visiting nurse service. The outcomes of ACC
by nurses and home-helpers indicated lower rates of
institutionalization and reduced waiting lists in the
study town, compared with the control town,
although control town rates of institutionalization did
not differ from all Japan rates (see article number 47).
3. Nursing into the community: Creating seamless
care
The third area, ‘nursing into the community’,refers to developing models to assist patients andfamilies when patients return home through multidisciplinary and collaborative discharge planning in hospitals in order to implement seamless care. To determine the effectiveness of newly established dischargeplanning service, several studies were conducted,
including the characteristics of patients referred fordischarge planning and of those identified being at
risk (see article number 35).
A quasi-experimental trial rexamined the effectiveness of early discharge planning for at-risk hospitalized elders. The findings indicated decreased anxiety
in patients and caregivers, and effectiveness in terms
of prevention of prolonged hospital stay (see article
number 36).
A book ‘Discharge Planning: A Challenge at theTokyo University Hospital’ was published in 2002, as
one of scholarly outcomes of this research activity.

Current Research Projects
Current research of the community health nursing
department is dedicated to further exploration ofways to promote health and continuation of projects
in the preceding years. To list a few are;1. Support program for the lonely elderly, fall prevention program, and demonstration projects such as
developing check list of unilateral spatial neglect
and related disorders amongst cerebro-vascularpatients in community settings.
2. Review and expansion of long-term care in a ruraltown, Town M, for the evaluation Long-Term Care
Insurance.
3. Seamless care between hospital and community 1)
Development and test of usefulness of check list333

for potential high-risk patients for referral of discharge planning support team; 2) Investigation ofdaily life problems immediately after discharge.

Future Plan
Since numbers of elderly in Japan are rapidly
increasing and family support is dwindling, thedepartment endeavors to take a lead role in research
related to establishing basic home care programs thatare efficient, acceptable by funding bodies (e.g. government) and adaptable by the local communities.
Therefore,
1. Faculty and graduate students of department arededicated to developing community interventions
specifically for care of demented and bed-ridden
elders.
2. Health promotion intervention studies to strengthen community competence and lines of resistancethrough programs, such as: 1) developing volunteer networking which offers day care programs toprevent elders from deterioration, and bed-ridden
or demented; 2) support programs for 75 years
and older by offering preventive home visiting 2-3
times a year; 3) strengthen and support community health nurses in developing skills and sustainable capacity-building developmental approaches
towards community partnerships and activities in
the community.

Research Grants (selected 5)
Sachiyo Murashima (P.I.), Katsuko Kanagawa, Hiromi
Kawagoe.
A study on organization and specification of functions of home care nursing: From the viewpoint of
24-hour care plan.
Grant-in-Aid for Scientific Research (B) by the
Ministry of Education and Culture in 1998-1999;￥12,400,000.
Sachiyo Murashima (P.I.), Shuhei Ryu, Masako Yamada,
Yutaka Tagami, Hiromi Watanabe, Naomi Sumi,
Naoko Takeuchi.
A study on promoting the model project of a combination type of nursing and home-help service.
Research Grant for Welfare of the Elderly by the
Ministry of Health, Labour and Welfare in 1999;￥5,000,000.
Sachiyo Murashima (P.I.), Tomoe Nogawa.
A research on promotion of well-being of the elderly
by public health nurses’ activities.
Research Grant for Welfare of the Elderly by the
Ministry of Health, Labour and Welfare: Research
Project for Supporting Independent and Healthy
Elders in 2000;￥10,000,000.
Hidetoshi Endo (P.I.), Sachiyo Murashima, Satoko
Nagata.

334

A study on effectiveness of the support system ofdischarge planning for hospitalized elders in special
functioning hospitals.
Research Grant for Longevity Sciences from the
Ministry of Health, Labour and Welfare in 20002002; ￥1,500,000 in each year.
Sachiyo Murashima (P.I.), Ryutaro Takahashi, Satoko
Nagata, Megumi Haruna.
Effectiveness of the program for discharge planningfor elderly in-patients.
Grant-in-Aid for Scientific Research (B) by the
Ministry of Education and Culture in 2001-2002;￥9,200,000.

Select Publications*: copy attached.
1. Murashima S and Iida S. A study on health behavior
and life style of primary school children: The difference between urban and rural communities, fifteen
years ago and now. Research Journal of School
Health 23(11), 531-539, 1981. (in Japanese)
2. Murashima S, Ryu S, Tanaka T and Tomari T. A
study on trends of the regional variations of physicalstandards in Japanese schoolchildren. Research
Journal of School Health 24(2), 89-97, 1982. (inJapanese)
3. Murashima S. A case study on family life of young
women with Diabetes Mellitus: How much can theirhusbands, relatives, neighborhood and children
understand their situations? Journal of Japan
Academy of Nursing Science 2(1), 39-47, 1982. (inJapanese)
4. Murashima S, Iida S, Ishii Y, Fujita A, Matsushita K
and Watanabe H. Health condition and home care of
the aged with 75 years and over: Regional differences
between urban and suburban areas. Bulletin of St.
Luke’s College of Nursing 15, 28-43, 1989. (inJapanese)
5. Murashima S, Iida S, Ishii Y, Matsushita K, Fujimura
M, Sanuki J, Hanazawa K, Watanabe H and Sakurai
N. Health conditions and home care of the frail elders with 75 years and over: Regional differences
between urban and suburban areas. Bulletin of St.
Luke’s College of Nursing 16, 60-69, 1990. (inJapanese)
6. Yoshio K, Murashima S and Iida S. A study onchanges of DM patients’ life style and the affecting
factors. Journal of Japan Academy of Nursing Science10(1), 24-36, 1990. (in Japanese)
7. Fukanoki T, Murashima S and Iida S. Analyses offactors affecting home oxygen therapy (HOT)patients’ Quality of Life: Focusing on their life satisfaction. Journal of Japan Academy of Nursing Science11(1), 9-21, 1991. (in Japanese)
8. Fukaya A, Murashima S and Iida S. Analyses of factors affecting the change in ADL of post-infarcthemiplegics elders living at home: Focusing on thedaily life style of the patients and their families.

Journal of Japan Academy of Nursing Science 11(2),44-54, 1991. (in Japanese)
9. Murashima S, Iida S, Matsushita K, Sakurai S andNaruki H. Building a care system for frail elderly inthe community. Annual Report: St. Luke’s College ofNursing, WHO Collaborating Center for Nursing andMidwifery Development in PHC 1990/1, 48-54, 1992.
10. Mizuno T, Murashima S and Iida S. Analyses on careburden and cognitive discrepancies of care role
between frail elders and their care-givers. Journal ofJapan Academy of Nursing Science 12(2), 17-29,
1992. (in Japanese)
11. Murashima S, Satoh R, Naruki H, Kurebayashi M and
Iida S. Issues in mobilizing the bedridden elderly athome. Annual Report: St. Luke’s College of Nursing,
WHO Collaborating Center for Nursing andMidwifery Development in PHC 1991/2, 8-20, 1993.
12. Nishio J, Murashima S and Iida S. Health behavior
model based on analysis of factors affecting alcohol
behavior of company employees. Japanese Journal of
Public Health 41(7), 607-617, 1994. (in Japanese)
13. Murashima S. Limitation of care-giving to the elders
by the families and the conditions for the society totake care of the elders. Research Journal of Care andWelfare 2(1), 23-30, 1995. (in Japanese)
14. Ota K, Tamura M, Sei M, Koyama M, Hishinuma M
and Murashima S. The content and its teaching
method of nursing for elderly in Baccalaureate
Program. Bulletin of St. Luke’s College of Nursing 21,
34-40, 1995. (in Japanese)
15. Murashima S, Murakami K, Kadokawa T, Sasaki M,
Naruki H, Kawakami H, Arai H, Iwanaga T and
Iwamuro S. Strategies of public health nurses topractice in rural areas and urban areas. Japanese
Journal of Public Health Nurses 51, 91-96, 1995. (inJapanese)
16. Murashima S, Yamada M, Tagami Y, Satoh M,
Uchida E, Miyazaki W, Kawagoe H, Hoshi T, Ooura
H, Horii T, Nakayama I and Yamazaki M. Factsthrough the project of around-the-clock nursing andcare provided by visiting nurse service stations.
Japanese Journal of Home Care Nursing 1(2), 83-92,
1996. (in Japanese)
17. Murashima S, Kim J and Fukui S. Outcomes in homecare. Japanese Journal of Nursing Research 30(5),351-363, 1997. (in Japanese)
18. Murashima S, Fukui S and Kim J. Several kinds ofrecording formats for home care developed on thebasis of outcome. Japanese Journal of Nursing
Research 30(5), 47-63, 1997. (in Japanese)
19. Okada M, Murashima S and Asahara K. A study oncompetencies used by public health nurses in creating new health care systems in the community.
Japanese Journal of Public Health 44(4), 309-321,
1997. (in Japanese)
20. Murashima S and Yamada M. Support for home carein the community: Principles of providing home help
and nursing care. Health Sciences 14(1), 6-11, 1998.
(in Japanese)

21. Murashima S. Role and issues of visiting nurse service in the aging society: For effective home care.
Japanese Journal of Gerontological Sociology 20(1),16-24, 1998. (in Japanese)
22*.Murashima S, Zerwekh JV, Yamada M and Tagami Y.
Around-the-clock nursing care for the elderly inJapan．Image: Journal of Nursing Scholarship 30(1),37-41, 1998．
23. Konishi K, Murashima S and Kanagawa K. A study
on stress-coping in the stroke rehabilitation process:A qualitative analysis on problems recognized bypatients. Japanese Journal of Home Care 1(1), 56-66,
1998. (in Japanese)
24. Okamoto E, Murashima S and Saito E. The significance of day care service for elders with dementia
and their caregivers: What is the difference between
the day with day care service and the day with noservice? Japanese Journal of Public Health 45(12),1152-1161, 1998. (in Japanese)
25*.Murashima S, Hatono Y, Whyte N and Asahara K.
Public health nursing in Japan: New opportunities
for health promotion. Public Health Nursing 16(2),133-139, 1999.
26. Murashima S. Function and role of nurses in aroundthe-clock in-home care service. Health Care 40(5),387-392, 1998. (in Japanese)
27. Murashima S, Ryu S, Asahara K, Yamada M and
Tagami Y. A system supporting the elderly at home:
Demand and effectiveness of 24 hour home carewhich includes medical treatment. GeriatricMedicine 36(12), 1789-1795, 1998. (in Japanese)
28. Yamawaki A and Murashima S. Home care service
utilization by severely mentally and physically handicapped persons. Japanese Journal of Public Health45(6), 499-511, 1998. (in Japanese)
29. Ito A, Murashima S, Choi J and Horii T. The meaningof the characteristics on the function of night-timenurses’ visits: Through the analyses of the users’
record of the visiting nurse station which provides
around-the-clock in-home care by three shifts ofnurses. Journal of Japan Academy of Home HealthCare 3(1), 68-77, 1999. (in Japanese)
30. Murashima S, Saito E, Hattori M, Hagiwara A, Sumi
N, Ueno M and Kanagawa K. Study on competenciesof community health nurses and introduction of theactivity by Dr. Joyce Zerwekh: Focused on home visits and visiting nurse service. The Japanese Journal of
Nursing Research 32(1), 3-14, 1999. (in Japanese)
31*.Murashima S, Asahara K, White MC and Ryu S. The
meaning of public health nursing: Creating 24 hourcare in a community in Japan. Nursing and Health
Sciences 1, 83-92, 1999.
32. Murashima S and Hollaender L. How do the people
in Denmark treat with elders with dementia. The
Japanese Journal of Occupational Therapy 34(5), 594597, 2000. (in Japanese)
33. Murashima S. Evaluation of home care service.
Health Care 42(4), 263-269, 1999. (in Japanese)
34*.Takeuchi N, Murashima S and Hattori M.
Comparison between two types of around-the-clock335

in-home care systems: A single type of home help
service and a combination type of nursing and home
help service, focused on the way of collaboration andits influence. Journal of Japan Academy of Home
Health Care 4(1), 24-30, 2000. (in Japanese)
35*.Murashima S, Nagata S, Toba K, Ouchi Y and Sagawa
Y. Characteristics of patients referred for dischargeplanning from a geriatric ward at a national university hospital in Japan: Implications for improving hospital programs. Nursing and Health Sciences 2, 153161, 2000.
36*.Sumi N, Murashima S, Toba K and Ouchi Y. The
effectiveness of early discharge planning for hospitalized elders who had expected difficulties of discharge: A quasi-experimental trial in a geriatric wardof a highly advanced medical center. Journal of theJapanese Society on Hospital Administration 38(1),29-40, 2001. (in Japanese)
37. Yokoyama A, Murashima S, Nagata S, Yanagisawa A,
Wakabayashi K, Tashiro T, Toba K and Ouchi Y.
Follow-up study of patients who received dischargeplanning by a special department in a national university hospital. Journal of the Japanese Society on
Hospital Administration 38(1), 53-61, 2001. (inJapanese)
38. Kayama M, Zerwekh J, Thornton KA and Murashima
S. Japanese expert public health nurses empower
clients with schizophrenia living in the community.
Journal of Psychosocial Nursing 39(2), 40-45, 2001.
39. Toba K, Akishita M, Tanaka S, Kato T, Kawai S,
Yamaguchi N, Murashima S, Iguchi A and Sasaki H.
Knowledge and utilization on the comprehensive
evaluation of functions and of physicians’ opinionsin Japan: Joint research based on a wide variety of
hospitals and facilities in Japan. Journal of Japan
Academy of Geriatric Medicine 38(2), 139-147, 2001.
(in Japanese)
40. Nagata S and Murashima S. Current situation andissues of support program for early discharge. HealthCare 44(2), 95-99, 2002. (in Japanese)
41. Murashima S, Nagata S and Haruna M. Visiting
nurse service: Transition from hospitals to visiting
nurse service stations. The Japanese Journal of
Nursing Research 35 (1), 15-24, 2002. (in Japanese)
42. Kim J and Murashima S. Comparison of the homecare system for the elderly in Japan and Korea:
Towards an advanced home care system. Journal of
Korean Academy of Nursing 31(7), 1166-1173, 2001.
43*.Asahara K, Momose Y, Murashima S, Okubo N and
Magilvy JK. The Relationship of social norms to useof services and caregiver burden in Japan. Journal of
Nursing Scholarship 33(4), 375-380, 2001.
44*.Murashima S, Nagata S, Magilvy JK, Fukui S andKayama M. Home care nursing in Japan: A challenge
for providing good care at home. Public HealthNursing 19(2), 94-103, 2002.
45. Asahara K, Momose Y and Murashima S. Familycare-giving of the elderly and Long-Term Care
Insurance in rural Japan. International Journal of
Nursing Practice 8, 167-172, 2002.

336

46. Honda A, Saito E, Kanagawa K and Murashima S.
The relation between characteristics and activities ofdaily living of the elderly living alone. Japanese
Journal of Public Health 49(8), 795-801, 2002. (inJapanese)
47*.Murashima S and Asahara K. The effectiveness of
the around-the-clock in-home care system: Did it
prevent the institutionalization of frail elderly? Public
Health Nursing 20(1): 13-24, 2002.
48*.Choi J, Murashima S, Hattori M, Horii T, Nagata S
and Asahara K. Factors influencing the utilization ofvisiting nursing and home-help services. Japanese
Journal of Public Health 49(9), 948-958, 2002. (inJapanese)
49. Murashima S, Yokoyama A, Nagata S and Asahara K.
The implementation of the Long-Term Care
Insurance in Japan: Focused on the trend of homecare. Home Health Care Management & Practice15(6), 2002 (in press).
50. Momose Y, Asahara K and Murashima S. A trial tosupport fam;;ily caregivers in the Long-Term Care
Insurance: Self-help groups in small communities.
Home Health Care Management & Practice (in press).

Department of International
Community Health
Outline and Research Objectives
The mission of our department is to seek equity and equality in health and health care,and social justice within and across the nations with caring, sharing and compassionate
spirit, by the people, with the people and for the people, by means of comprehensive andintegrated scientific research and education on health and health care, and social development in collaboration with other related institutes of various disciplines including universities, governmental and non-governmental organizations and private sectors.
Our aim is to bring together the clinical, public health and social science research toaddress the broad issues of health and health care.
The goals are: 1) to investigate how to change the status quo by improving health statusof the most vulnerable, the most underprivileged and the poorest of the poor; 2) to undertake research on the influences of ‘globalization’ and ‘free market’ system on health andsocial development; 3) to investigate the mechanisms to reduce inequalities between andwithin nations on health and development and 4) to develop the mechanisms to link theabove three goals with individual research activities described below.
Focusing on the followings:
1. Target diseases or topics: 1) HIV/AIDS (prevention and health care services), 2)
Tuberculosis control, 3) Leprosy control, 4) Appropriate treatment and prevention ofcommon diseases and injuries including traffic accidents in Japan and developing countries, 5) Public health aspects of Stroke, 6) Health care services for international migrant
workers in Japan, 7) Maternal and Child Health Care (issues on contraceptives excluded).
2. Site/Field: 1) Aral Sea area, 2) Cambodia, 3) Bangladesh, 4) Nepal, 5) Nicaragua and 5)
Japan.
3. Research topics are related to Epidemiology, prevention and health care service delivery.

Faculties and Students
Professor and Chair Susumu Wakai, MD, PhD (1999-)
Lecturer
Nasamine Jimba, MD, PhD
Associate .................................2
Graduate student ..................20
Research student.....................1
Secretary .................................2

Past Research and Major Accomplishments
Please refer to the Select Publications related to
Research on International Health including Advocacy
papers (number 1 to 31)

Current Research
1) Injury prevention in Japan and developing countries2) HIV/AIDS prevention and care among migrant
workers living in Japan
3) Leprosy and tuberculosis control in Asian countries

4) Environmental degradation and people’s health:Aral Sea region and elsewhere
5) Epidemiology of Stroke and its prevention
6) Maternal and child healthIn each six research field a concerted effort has
been made with the Faculty members and Graduate
students to contribute for improving people’s healthstatus in developing countries but in developed countries including Japan as well.

Future Prospects
We continue focusing our research efforts on the 6
major research fields described above. We utilize deferent tools such as Epidemiological, medical.
Quantitative and Qualitative research methods. We
also need human resource development for continuing such research efforts by mobilizing Graduate students.

337

Research Grants
Ministry of Education and Science
1. FY 1999 5.5 million Yen: S Wakai, K Pyo.
Mechanisms of spondylotic changes of the spine.
2. FY 2001~2002: 3.4 million Yen: S Yanagisawa andS Wakai. Health reform in Cambodia and its
effects on people’s health seeking behavior
3. FY 2002-2003: 8 million Yen: J Okumura and S
Wakai Ministry of Health and Welfare
4. FY 2000-2002: 21.5 million Yen: S Wakai. Healthsystem building for supporting people with
HIV/AIDS from overseas.

Select Publications
Research on International Health including
Advocacy paper
1. Wakai S: Primary health care projects and social
development. Lancet 345:1241, 1995
2. Wakai S: Sanctions against Iraq. Lancet 347: 200,
1996
3. Wakai S: Access to the Internet. Science 271: 1347,
1996
4. Kunii O, Wakai S, Honda T, Tsujimoto K: Role ofexternal medical volunteers after disasters. Lancet
347: 1411, 1996
5. Wakai S. Role of neurosurgeons in Japan. Lancet 349:
140, 1997
6. Yoshimoto Y, Wakai S: Unruptured intracranial vertebral artery dissection: clinical course and serial radiographic imagings. Stroke 28:370-374, 1997
7. Wakai S: Global health care and neurosurgeons.
Neurol Med Chir 37:483-490, 1997
8. Wakai S: Scourge of Japanese encephalitis in southwestern Nepal. Lancet 351:759, 1998
9. Wakai S: Health care in Burma. Lancet 352: 1230,
1998
10. Wakai S: Surgical treatment for incidentally discovered intracranial aneurysms. Lancet 1999; 353:19751976
11. Barua S, Wakai S, Shwe T, Umenai T: Leprosy elimination through integrated basic health services in
Myanmar: the role of midwives in the pre- and postelimination phases. Leprosy Rev 70:174-179, 1999
12. Yoshimoto Y, Wakai S: Cost-effectiveness analysis ofscreening for asymptomatic, unruptured intracranial
aneurysms: A mathematical model. Stroke 30: 16211627 1999
13. Inaoka E, Wakai S, Nakamura Y, Babily YA,
Saghayroun AAR. Correlate of visit regularity amongfamily planning clients in urban Yemen. Adv
Contraception 1999; 15: 257-274
14. Wakai S. Pysicians for peace. Lancet 2000; 355: 13651366
15. Wakai S. Life after sanctions: the fate of Iraq. Lancet
2000; 356: 685

338

16. Kuroiwa C, Vongphrachanh P, Chosa T, Murakami
H, Hashizume M, Wakai S, Tanaka M. Risk ofpoliomyelitis importation and reemergence in Laos.
Lancet 2000; 356: 1487-88.
17. Toyama N, Wakai S, Nakamura Y, Andryansyah
Arifin. Mother’s working status and nutritional status of children under the age of five in urban lowincome community, Surabaya, Indonesia. J Trop
Pedatr 2001; 47: 179-181
18. Jimba M, Wakai S. Is Gandhi’s philosophy of nonviolence dead in Nepal? Lancet 2001:358;1018- 2.
19. Nakahara S, Wakai S. Underreporting of traffic
injuries involving children in Japan. Injury
Prevention 2001;7: 242-244
20. Sase E, Wakai S. A world war against terrorism.
Lancet 2001:358;1365
21. Ichikawa M, Nakahara S, Wakai S. Death withunbeled passengers. Lancet 2002:359;43-44
22. Ichikawa M, Okumura J, Wakai S. Two decisions ofJapanese court on detaind Afghan asylum seekers.
Lancet 2002:359;448
23. Saengdidtha B, Lapparat G, Torugsa K, Suppadit W,
Wakai S. Sexual Behavior and HIVInfection Among
Thai Army Conscripts between 1992 and 1998.
Military Med 2002: 167(4):272-6
24. Nakahara S, Wakai S. Differences between Japanesepre-school and school-age pedestrian mortality andmoridity trends. Public Health 2002:116;1-7
25. Kunii O, Nakamura S, Abdur R, Wakai S. The Impact
on Health and Risk Factors of the Diarrhoea
Epidemics in the 1998 Bangladesh Floods. Public
Health 2002:116; 68-74
26. Okumura J, Wakai S, Umenai T. Drug utilisation
practice at household focusing self-medication inrural communities in Vietnam. Soc Sci Med
2002:54;1875-1886
27. Md. Akramul Islam, Susumu Wakai, Nobukatsu
Ishikawa, A.M.R. Chowdhury, J. Patrick Vaughan:
Tuberculosis control by community health workers
inBangladesh: are they more cost-effective? Bull
WHO 2002:80;445-450
28. Phoxay C, Okumura J, Nakamura Y, Wakai S.
Influence of Women’s Knowledge and HealthSeeking Behaviour on Maternal Health Care
Utilization in Southern Laos. Asia-Pacific J Public
Health 2001:13;13-18
29. Flores FP, Umenai T, Wakai S. Shoud communitymanaged drugstores be phased out? Asia-Pacific J
Public Health 2001:13;9-12
30. Wakai S. Mobilization of Cuban doctors in developing countries. Lancet 2002: 360;92
31. Miura S, Kunii O, Wakai S. Home Gardening in
Urban Poor Communities of the Philippines. Int J
Food Sci Nutrition 2002 (in print)
32. M Hashizume,1 O Kunii,1 S Sasaki,2 T Shimoda,3 S
Wakai1 and M Chiba. Anemia and Anthropometrical
Status of School-age Children in the Aral Sea Region,
Kazakhstan. J Trop Pediatr (in press)

Clinical Research and Epidemiology
33. Wakai S, Fukushima T, Furihata T, Sano K:
Association of cerebral aneurysm with pituitary adenoma. Surg Neurol 12:503-507, 1979
34. Wakai S, Fukushima T, Teramoto A, Sano K:
Pituitary apoplexy: its incidence and clinical significance. J Neurosurg 55:187-193, 1981
35. Sano K, Wakai S, Ochiai C, Takakura K:
Characteristics of intracranial meningiomas in childhood. Child’ Brain 8:98-106, 1981
36. Wakai S, Yamakawa K, Mananka S, Takakura K:
Spontaneous intracranial hemorrhage caused bybrain tumor: its incidence and clinical significance.
Neurosurgery 10:437-444, 1982
37. Segawa H, Wakai S, Tamura K, Yoshimasu N,
Kakamura O, Ohta M: Computed tomographic measurement of local cerebral blood flow by Xenon
enhancement. Stroke 14:356-362, 1983
38. Wakai S, Narita J, Hashimoto K, Nagai M: Dermoid
cyst communicating with the subarachnoid spaceand lateral ventricle: demonstration by metrizamide
computed
tomographic
cisternography.
Neurosurgery 13:696-698, 1983
39. Wakai S, Arai T, Nagai M: Congenital brain tumors.
Surg Neurol 21:597-609, 1984
40. Wakai S, Nagai M: Ventricular diverticulum. J Neurol
Neurosurg Psychiatry 47:514-517, 1984
41. Wakai S, Ueda Y, Inoh S, Nagai M: Angiographicallyoccult angiomas: A report of thirteen cases withanalysis of the cases documented in the literature.
Neurosurgery 17:549-556, 1985
42. Wakai S, Nagai M: Histological verification of
microaneurysms as a cause of cerebral haemorrhage
in surgical specimens. J Neurol Neurosurg Psychiatry
52:595-599, 1989
43. Wakai S, Hashimoto K, Watanabe N, Inoh S, Ochiai
C, Nagai M: Efficacy of closed-drainage system intreating chronic subdural hematoma: A prospective
comparative study. Neurosurgery 26:771-773, 1990
44. Iwanaga H, Wakai S, Ochiai C, Narita J, Inoh S, Nagai
M: Ruptured cerebral aneurysms missed by initialangiography. Neurosurgery 27:45-51, 1990
45. Wakai S, Kumakura N, Nagai M: Lobar intracerebral
hemorrhage: A clinical, radiographic and pathologic
study of 29 consecutive operated cases with negative
angiography. J Neurosurg 76: 231-238, 1992

extracellular space of the brain. Neurosurgery
18:548-554, 1986
50. Wakai S, Meiselman SE, Brightman MW: Focal circumvention of blood-brain barrier with grafts ofmuscle, skin and autonomic ganglia. Brain Res
386:209-222, 1986

Biomedical Research
46. Wakai S, Hirokawa N: Development of the bloodbrain barrier to horseradish peroxidase in the chickembryo. Cell Tiss Res 195:195-203, 1978
47. Wakai S, Matsutani M, Mizutani H, Sano K: Tight
junctions in choroid plexus papillomas. Acta
Neuropathol 45:159-160, 1979
48. Wakai S, Hirokawa N: Development of blood-cerebrospinal fluid barrier to horseradish peroxidase in
the avian choroidal epithelium. Cell Tiss Res
214:271-278, 1981
49. Wakai S, Meiselman SE, Brightman MW: Muscle
grafts as entries for blood-borne proteins into the339

Department of
Human Genetics
Outline and Research Objectives
The Department of Human Genetics was established in April 1992 as one of the departments at the newly established School of International Health. The department educates
students at the Graduate School of International Health with courses covering basic principles as well as clinical applications of human genetics. A series of lectures on human genetics are also provided to the first year students at the School of Medicine (compulsory). In
addition, for undergraduate students at the School of Health Sciences, a series of lectures
are given to each of the sophomore (Human Genetics I, compulsory) and junior (Human
Genetics II, elective) classes.
The major research projects in the department are: (1) Search for susceptibility genes torheumatic diseases and other autoimmune diseases, (2) Search for susceptibility genes toclinical subsets of malaria, (3) Genome diversity of Asia-Pacific populations, (4) Search forsusceptibility genes to behavioral disorders, and (5) Analysis of gene expression profiles in
inflammatory tissues and cell differentiation.

Faculties and Students
Professor and Chair Katsushi TOKUNAGA, DSc
Associate Professors Naoyuki TSUCHIYA, MD, DMSc
Associate
Jun OHASHI, DHSc
Postdoctoral Fellow ................2
Graduate student ..................12
Research student.....................3
Research Technician ...............7
Secretary .................................2

Past Research and Major Accomplishments(1) By means of extensive variation screening of anumber of candidate genes followed by association analyses with complex diseases including
rheumatic diseases, malaria and narcolepsy, several disease-gene associations have been newly
found as follows.
- Rheumatoid arthritis (RA): LIR1(ILT2),
BLYS/BAFF-R, TNFR2, FCGR3A/HLA-DRB1- Systemic lupus erythematosus (SLE): TNFR2,
CD19, FCGR2B, FCGR3A
Cerebral malaria (in Thailand): HLA-B, TNFA, CD36
Severe malaria (in Thailand): HLA-DRB1, iNOS
Narcolepsy: TNFA, TNFR2
ANCA-associated vasculitis: HLA-DRB1(2) Studies on HLA gene and haplotype distribution in
East Asian and Pacific populations have demonstrated genetic affinities and clustering of the populations, multiple migration and dispersal routesof ancestral populations, and the genetic link
between Asians and Native Americans.

340(3) Gene expression profiling in synovia from RA
patients demonstrated several up-regulated genes
including ID family genes. Also a number of upregulated and down-regulated genes including
TNF-TNFR family were identified in the differentiation of monocytes into dendritic cells.

Current Research(1) Establishment of Human SNP (single nucleotide
polymorphism) Typing Center for large scale SNP
typing (goal: 1,000,000 typing/year).
(2) Continue the search for susceptibility genes torheumatic diseases, malaria subsets, and narcolepsy mainly by means of candidate gene approach.
(3) Screening of novel candidate regions for susceptibility genes to SLE and narcolepsy by means ofwhole genome association studies.
(4) Analyses of linkage disequilibrium blocks on chromosome 1 for the determination of primary genes.
(5) Functional analyses of disease-associated SNPs andID family genes.

Future Prospects(1) High-throughput SNP typing in the Human SNP
Typing Center will contribute to the identificationof susceptibility genes to various complex diseases
including allergy, diabetes, hypertension, andinfectious diseases.
(2) Using the new strategy of the whole genome association analyses, several candidate regions for SLE
and narcolepsy will be newly found.

(3) Extensive studies on linkage disequilibrium blocks
will contribute to effective search of disease associated genes.
(4) Further development of new high-throughput SNP
typing technologies.

Research Grants<Only the grants of more than 10,000,000
yen/year are listed below>
1. “Medical Genome Science”, Grant-in-Aid for
Scientific Research on Priority Areas, Grant-in-Aid
for Scientific Research by The Ministry of Education,
Culture, Sports, Science and Technology: Human
SNP Typing Center. 2001-2004.
2. “Medical Genome Science”, Grant-in Aid for
Scientific Research on Priority Areas, Grant-in Aid
for Scientific Research by The Ministry of Education,
Culture, Sports, Science and Technology:
Susceptibility genes to rheumatic diseases.
28,000,000 yen/year. 2000-2004.
3. “Genome Informatics Research and Technology
Development”, by the New Energy and Industrial
Technology Development Organization (NEDO):Development of DNA capillary array. 10,000,000
yen/year. 1999-2002.
4. “Genome Diversity Project”, by the New Energy andIndustrial Technology Development Organization(NEDO): Genome-wide association studies onrheumatoid arthritis. 10,000,000 yen/year. 20012004.

Select Publications
1. Tokunaga K, Araki C, Juji T, and Omoto K: Genetic
polymorphism of the complement C2 in Japanese.
Hum. Genet. 58: 213-216, 1981.
2. Tokunaga K, Omoto K, Juji T, and Itoh T:
Polymorphism of BF, C2 and GLO in Japanesepatients with insulin-dependent diabetes mellitus:
Confirmation of an increase of BF*FT. Hum. Genet.
62: 86-88, 1982.
3. Tokunaga K, Yukiyama Y, and Omoto K:
Polymorphism of the complement C6 in Japanese. J.
Immunogenet. 10: 419-424, 1983.
4. Tokunaga K, Omoto K, Yukiyama Y, Sakurai M, SajiH, and Maruya E: Further study on a BF silent allele.
Hum. Genet. 67: 449-451, 1984.
5. Matsuki K, Juji T, Tokunaga K, Naohara T, Satake M,
and Honda Y: Human histocompatibility leukocyte
antigen (HLA) haplotype frequencies estimated fromthe data on HLA class I, II, and III antigens in 111
Japanese narcoleptics. J. Clin. Invest. 76: 2078-2083,
1985.
6. Tokunaga K, Dewald G, Omoto K, and Juji T: Family
study on the polymorphisms of the sixth and seventh components (C6 and C7) of human complement: Linkage and haplotype analyses. Am. J. Hum.
Genet. 39: 414-419, 1986.

7. Matsuki k, Juji T, Tokunaga K, Mochizuki M,
Hayashi K, Fujino Y, Numaga J, and Yamashita H:
HLA antigens in Behcet’s disease with refractoryattacks. Tissue Antigens 29: 208-213, 1987.
8. Tokunaga K, Sauerecker G, Kay P, Christiansen F,
Anand R, and Dawkins RL: Extensive deletions andinsertions in different MHC supratypes detected bypulsed field gel electrophoresis. J. Exp. Med. 168:933-940, 1988.
9. Watanabe Y, Tokunaga K, Matsuki K, Takeuchi F,
Matsuta K, Maeda H, Omoto K, and Juji T: Putative
amino acid sequence of HLA-DRB chain contributing
to rheumatoid arthritis susceptibility. J. Exp. Med.
169: 2263-2268, 1989.
10. Matsuki K, Juji T, Tokunaga K, Takamizawa M,
Maeda H, Soda M, Nomura Y, and Segawa M: HLA
antigens in Japanese patients with myasthenia
gravis. J. Clin. Invest. 86: 392-399, 1990.
11. Tokunaga K, Jin F, Mauff G, Nishimukai H, Simeoni
E, Suzuki K, Weber W: C6 reference typing report.
Complement Inflamm. 7: 234-239, 1990.
12. Kuwata S, Tokunaga K, Jin F, Juji T, Sasaki T, andHonda Y: Analysis of HLA-DRB gene in narcolepsy.
N. Engl. J. Med. 324: 271-272, 1991.
13. Tokunaga K, Zhang WJ, Christiansen FT, andDawkins RL: The genomic structure of two ancestral
haplotypes carrying C4A duplications. Immunogenet.
34: 247-251, 1991.
14. Uchigata Y, Kuwata S, Tokunaga K, Eguchi Y,
Takayama-Hasumi S, Miyamoto M, Omori Y, Juji T,
and Hirata Y: Strong association of insulin autoimmune syndrome with HLA-DR4. Lancet 339: 393394, 1992.
15. Mitsunaga S, Kuwata S, Tokunaga K, Uchikawa C,
Takahashi K, Akaza T, Mitomi Y, and Juji T: Family
study on HLA-DPB1 polymorphism: Linkage analysis
with HLA-DR/DQ and two “new” alleles. Hum.
Immunol. 34: 203-211, 1992.
16. Bannai M, Mazda T, Tokunaga K, and Juji T: DNA
single-strand conformation polymorphism method
to distinguish DR4 alleles. Lancet 341: 769, 1993.
17. Lin L, Tokunaga K, Ogawa A, Ishikawa Y, Kashiwase
K, Akaza T, Tadokoro K, and Juji T: Genotyping andassociation analysis of HLA-B61 in Japanese. Hum.
Immunol. 37: 95-100, 1993.
18. Wang L, Juji T, Tokunaga K, Takahashi K, Kuwata S,
Uchida S, Tadokoro K, and Takai K: Polymorphic
microsatellite markers for the diagnosis of graft-versus-host disease. N. Engl. J. Med. 330: 398-401, 1994.
19. Oka T, Matsunaga H, Tokunaga K, Mitsunaga S, JujiT, and Yamane A: A simple method for detecting
single base substitutions and its application to HLADPB1 typing. Nucleic Acid Research 22: 1541-1547,
1994.
20. Tokunaga K, Sideltseva EW, Tanaka H, Uchikawa C,
Nieda M, Sideltsev VV, Zhuravleva E, Imanishi T,
Itoh K, Akaza T, Takahashi K, Khalturin V, Alexeev
LP, and Juji T: Distribution of HLA antigens and haplotypes in the Buryat population in Siberia. Tissue
Antigens 45: 98-102, 1995.

341

21. Yoshida M, Ohtsuka R, Nakazawa M, Juji T, andTokunaga K: HLA-DRB1 frequencies of nonAustronesian-speaking Gidra in South New Guinea
and their genetic affinities with Oceanian populations. Am. J. Phys. Anthrop. 96: 177-181, 1995.
22. Uchigata Y, Tokunaga K, Nepom G, Bannai M,
Kuwata S, Dozio N, Benson EA, Ronningen KS,
Spinas GA, Tadokoro K, Hirata Y, Juji T, and Omori
Y: Differential immunogenetic determinants of polyclonal insulin autoimmune syndrome (Hirata’s
Disease) and monoclonal insulin autoimmune syndrome. Diabetes 44: 1227-1232, 1995.
23. Suto Y, Tokunaga K, Watanabe Y, and Hirai M:
Visual demonstration of the organization of thehuman complement C4 and 21-hydroxylase genes byhigh-resolution fluorescence in situ hybridization.
Genomics 33: 321-324, 1996.
24. Lin L, Tokunaga K, Tanaka H, Nakajima F, Imanishi
T, Kashiwase K, Bannai M, Mizuno S, Akaza T,
Tadokoro K, Shibata Y, and Juji T: Further moleculardiversity in the HLA-B15 group. Tissue Antigens 47:
265-274, 1996.
25. Ogasawara K, Yabe R, Uchikawa M, Saitou N, Bannai
M, Nakata K, Takenaka M, Fujisawa K, Ishikawa Y,
Juji T and Tokunaga K: Molecular genetic analysis ofvariant phenotypes of the ABO blood group system.
Blood 88: 2732-2737, 1996.
26. Fujiwara K, Isa K, Oka T, Maekawajiri S, Yamase A,
Akaza T, Tadokoro K, Juji T, Shibata Y, andTokunaga K: Large-scale DNA typing for humanplatelet alloantigens by PCR-PHFA (preferential
homoduplex formation assay). Brit. J.Haematol. 95:
198-203, 1996.
27. Bannai M, Tokunaga K, Tanaka H, Lin L, Kashiwase
K, Tokunaga K, and Juji T: Five HLA-B22 group alleles in Japanese. Tissue Antigens 49: 376-382, 1997.
28. Miyake M, Nakahori Y, Matsushita I, Kobayashi K,
Mizuno K, Hirai M, Kanazawa I, Nakagome Y,
Tokunaga K, and Toda T: YAC and cosmid contigs
encompassing the Fukuyama-type congenital muscular dystrophy (FCMD) candidate region on 9q31.
Genomics 40: 284-293, 1997.
29. Watanabe Y, Tokunaga K, Geraghty DE, Tadokoro K,
and Juji T: Large scale comparative mapping of MHC
class I region of predominant haplotypes in Japanese.
Immunogenet. 46: 135-141, 1997.
30. Tokunaga K, Ishikawa Y, Ogawa A, Wang H,
Mitsunaga S, Moriyama S, Lin L, Bannai M,
Watanabe Y, Kashiwase K, Tanaka H, Akaza T,
Tadokoro K, and Juji T: Sequence-based association
analysis of HLA class I and II alleles in Japanese supports conservation of common haplotypes.
Immunogenet. 46: 199-205, 1997.
31. Tsuchiya N, Shiota M, Moriyama S, Ogawa A,
Komatsu-Wakui M, Mitsui H, Geraghty ED, andTokunaga K: MICA allele typing of HLA-B27 positive
Japanese patients with seronegative spondylarthropathies and healthy individuals: Differential
linkage disequilibrium with HLA-B27 subtypes.
Arthritis Rheum. 41: 68-73, 1998.

342

32. Ogawa A, Tokunaga K, Lin L, Kashiwase K, Tanaka
H, Herrero MJ, Vilches C,Park MH, Jia GJ, Chimge NO, Sideltseva EW, Ishikawa Y, Akaza T, Tadokoro K,
and Juji T: Diversity of HLA-B61 alleles and haplotypes in East Asians and Spanish Gypsies. Tissue
Antigens 51:356-366, 1998.
33. Kobayashi K, Nakahori Y, Miyake M, Matsumura K,
Kondo-lida E, Nomura Y, Segawa M, Yoshioka M,
Saito K, Osawa M, Hamano K, Sakakibara Y, Nonaka
I, Nakagome Y, Kanazawa I, Nakamura Y, Tokunaga
K, and Toda T: An ancient retrotransposal insertion
causes Fukuyama-type congenital muscular dystrophy. Nature 394: 388-392, 1998.
34. Wuerzer R, Witzel-Schloemp K, Tokunaga K, Fernie
BA, Hobart MJ, and Orren A: Reference typing reportfor complement components C6, C7, and C9 including mutations leading to deficiencies. Exp. Clin.
Immunogenet. 15: 268-285, 1998.
35. Komata T, Tsuchiya N, Matsushita M, Hagiwara K,
and Tokunaga K: Association of tumor necrosis factor receptor 2 (TNFR2) polymorphism with susceptibility to systemic lupus erythematosus. Tissue
Antigens 53(6): 527-533, 1999.
36. Komatsu-Wakui M, Tokunaga K, Ishikawa Y,
Kashiwase K, Moriyama S, Tsuchiya N, Ando H,
Shiina T, Geraghty DE, Inoko H, and Juji T: MIC-A
polymorphism in Japanese and a MIC-A-MIC-B null
haplotype. Immunogenet. 49(7/8): 620-628, 1999.
37. Hatta Y, Ohashi J, Imanishi T, Kamiyama H, Iha M,
Shimabukuro T, Ogawa A, Tanaka H, Akaza T,
Gojobori T, Juji T, and Tokunaga K: HLA genes andhaplotypes in Ryukyuan suggest a recent gene flow
to the Okinawa islands. Hum. Biol. 71(3): 353-365,
1999.
38. Chida S, Hohjoh H, and Tokunaga K: Molecularanalyses of the possible RNA-binding protein genelocated in the human leukocyte antigen (HLA)-DR
subregion. Gene 240: 125-132 1999.
39. Bannai M, Ohashi J, Harihara S, Takahashi Y, Juji T,
Omoto K, and Tokunaga K: Analysis of HLA genes
and haplotypes in Ainu (from Hokkaido, northern
Japan) supports the premise that they descent from
Upper Paleolithic populations of East Asia. Tissue
Antigens 55 (2):128 -139, 2000.
40. Keicho N, Ohashi J, Tamiya G, Nakata K, Taguchi Y,
Azuma A, Ohishi N, Emi M, Park MH, Inoko H,
Tokunaga K, and Kudoh S: Fine localization of amajor disease-susceptibility locus for diffuse panbronchiolitis. Am. J. Hum. Genet. 66 (2): 501-507,
2000.
41. Shibue T, Tsuchiya N, Komata T, Matsushita M,
Shiota M, Ohashi J, Wakui M, Matsuta K, andTokunaga K: Tumor necrosis factorα 5’-flanking
region, TNF receptor ll, and HLA-DRB1 polymorphisms in the Japanese patients with rheumatoid
arthritis. Arthritis Rheum. 43 (4): 753-757, 2000.
42. Ohashi J, Yamamoto S, Tsuchiya N, Hatta Y, Komata
T, Matsushita M, and Tokunaga K: Comparison of
statistical power between 2×2 allele frequency andallele positivity tables in case-control studies of com-plex diseases genes. Ann. Hum. Genet. 65(2): 197206, 2001.
43. Ogasawara K, Yabe R, Uchikawa M, Nakata K,
Watanabe J, Takahashi Y, and Tokunaga K:
Recombination and gene conversion-like events may
contribute to ABO gene diversity causing various
phenotypes. Immunogenet. 53(3): 190-199, 2001.
44. Sakurai D, Yamaguchi A, Tsuchiya N, Yamamoto K,
and Tokunaga K: Expression of ID family genes inthe synovia from patients with rheumatoid arthritis.
Biochem. Biophys. Res. Commun. 284(2): 436-442,
2001.
45. Lapteva N, Ando Y, Nieda M, Hohjoh H, Okai M,
Kikuchi A, Dymshits G, Ishikawa Y, Juji T, andTokunaga K. Profiling of genes expressed in humanmonocytes and monocyte-derived dendritic cells
using cDNA expression array. Br. J. Haematol.
114(1): 191-197, 2001.
46. Tokunaga K, Ohashi J, Bannai M, and Juji T: Genetic
link between Asians and Native Americans:
Evidence from HLA genes and haplotypes. Hum.
Immunol. 62(9): 1001-1008, 2001.
47. Sirikong M, Tsuchiya N, Chandanayingyong D,
Bejrachandra S, Suthipinittharm S, Luangtrakool K,
Srinak D, Thongpradit R, Siriboonrit U, andTokunaga K: Association of HLA-DRB1*1502DQB1*0501 haplotype with susceptibility to systemic lupus erythematosus in Thais. Tissue Antigens
59 (2): 113-117, 2002.
48. Kyogoku C, Dijstelboem M.H, Tsuchiya N, Hatta Y,
Kato H, Yamaguchi A, Fukazawa T, Jansen D.M,
Hashimoto H, Winkel G.J. J, Kallenberg G.M. C, andTokunaga K: Fcg receptor gene polymorphisms inJapanese patients with systemic lupus erythematosus: Contribution of FCGR2B to genetic susceptibility. Arthritis Rheum. 46(5): 1242-1254, 2002.
49. Ohashi J, Naka I, Patarapotikul J, Hananantachai H,
Looareesuwan S, and Tokunaga K: Significant association of longer forms of CCTTT microsatellite repeat
in inducible nitric oxide synthase (iNOS) promoterwith severe malaria in Thailand. J. Infect. Dis.186:
578-581, 2002.
50. Kuroki K, Tsuchiya N, Tsao B, Grossman J, Fukazawa
T, Hagiwara T, Kano H, Takazoe M, Iwata T,
Hashimoto H, and Tokunaga K: Polymorphisms ofhuman CD19 gene: Possible association with susceptibility to systemic lupus erythematosus in Japanese.
Genes. Immun. (in press)(Number of the other original papers in English: 229)

343

Department of Developmental
Medical Sciences
Outline and Research Objectives
In 1996, the Department of Mother and Child Health was formed at the University ofTokyo. In Japan, it was the first division in International Health, which specifically focusedon maternal and child health issues. Maternal and child health involves all activities tomaintain and promote psychological and physiological health of mothers and children. The
field of maternal and child health covers (1) countermeasure for infectious diseases, (2)maternal and child nutrition, (3) prevention of diseases, and (4) protection of mothers andchildren from environmental unhealthy factors.
The study of maternal and child health not only covers research areas where clinicalintervention is possible but also it is beneficial for studying diseases where intervention isdifficult and abnormalities cannot be prevented. Maternal and child health research helps
administration and health policy makers to develop policies and to take decisions that are
in accordance with the current problems and are in the best interest of the communities.
Since 1998, in order to broaden its vision in research, scope of work and teaching activities
the division changed its name to “Developmental Medical Sciences”. Aging, human andenvironment, and their related items are newly added to our theme. This comes underInternational Biomedical Sciences, which is a division of Institute of International Health.
Since then, our research scope and work have expanded to foreign countries, mainly in Asia

Faculties and Students
Professor

Hiroshi Ushijima, M.D., Ph.D.
(1995 - to date)
Associate Professor Hideoki Fukuoka, M.D., Ph.D.
Associate .................................2
Postdoctoral Fellow ................3
Graduate student ..................20
Research student.....................4
Secretary .................................2

Past Research and Major Accomplishments
Background
After graduation from medical university andobtaining license of medical doctor, I first worked at
Tokyo University Hospital and related hospitals as apediatrician. At the same time, I studied clinicalresearch regarding pediatric infection, pediatricimmunology, and pediatric neurology. My research
produced results, which were related to clinical medicine. Following that I had an opportunity to study in
the United States as a virologist (Research fellow) atState University of Alabama for about 2 years. My
major was molecular study of Bunyaviruses, which
infect humans as well as animals. Some of the viral344

strains caused emerging diseases. During my research,
I discovered reassortant viruses and non-structure
proteins in the family. After returning to Japan, in
addition to being a pediatrician I also worked as a
researcher on molecular epidemiology, diagnosis anddevelopment of medicine for diarrhea caused by viruses and HIV at National Institute of Infectious Diseases(NIID) and National Institute of Public Health (NIPH)
as a chief and director, respectively. After that, I
moved to the University of Tokyo in 1995. Since then,I have been engaged in various researches in topics
ranging from infection, nutrition, maternal and childhealth in the world, with a special focus in Asia.
Followings are few researches, which I had carriedout and supervised in the University of Tokyo. The
researches include basic and field studies.
Diarrheal viruses
Rotavirus disease is an important diarrheal disease
especially in children. More than 500,000 children die
every year especially in developing countries. In thedeveloped countries, almost all infants have viral disease within 2 years after birth. My experience inworking on molecular epidemiology of rotavirus in
Japan expands over a period of a little more than 20
years. Besides, in the past four years, extensive collaboration has been done with researchers, microbiolo-

gists and pediatricians in countries like Korea, China,
Vietnam and Thailand. It has been done with similar
themes of epidemiological studies of diarrheal viruses.
These studies are important to develop vaccines
because serotypes and genotypes sometimes change
and the most appropriate vaccines are needed.
Major diarrheal viruses are rotavirus, astrovirus,
adenovirus, Norwalk virus and Sapporo virus.
Currently, we study all of them up to molecular level.
Immunodeficient persons discharge the viruses forlong time. Through research we have highlighted
complications of encephalopathy, hepatitis and others, which were caused by these viruses. Stool samples, slightly over 10,000 were collected for more than
20 years in five areas in Japan. Besides Japan, stool
samples were also collected in Asian countries. In
complicated cases, cerebrospinal fluids, bloods andliver tissues are also examined at times.
I have the honor of pioneering the concept anddeveloping molecular epidemiology of diarrheal viruses in Japan. During the process, I established particulars about diarrheal viruses causing encephalopathy
and other complications in humans. Due to seasonalchanges and serotype distribution, detail analyses of
all the viruses were done during the past 20 years.
Reassortment and recombination were also discovered. The changes in serotypes or genotypes wereaffected principally by time and also by geographic
distributions across the countries. We established thefacts about Norwalk virus spreading in early winterand rotavirus spreading in late winter in Japan.
However the detail of mode of spread of viruses isstill largely unknown, except for outbreaks of foodpoisoning and outbreaks at institutes. We alsobrought forward the fact that rotavirus infection
starts at the same time all over Japan. We have established a research system in Japan and nowadays weare examining the shellfishes and sewerages regularly.
The results of the research will contribute to thehealth policy and decision-making in Japan.
We newly developed RT-PCR, enzyme immunoassay, immunochromatography and latex agglutination
tests for rapid, easy and sensitive diagnosis. Anti-diarrheal virus agents are examined by in vitro assay. We
found that tea extracts, pine seed extracts and poly IC
were effective on rotavirus infection. This method isuseful to find anti-viral agents in vitro. In vivo assays
by using mouse model are also used. Mouse pups
born to and nursed by dams fed Bifidobacterium
breve and immunized orally with rotavirus were morestrongly protected against rotavirus-induced diarrhea
than those born to and nursed by dams immunizedwith rotavirus only.

tant areas of research not only in Asia in general but
also in Japan in particular. It is now an establishedfact that HIV, hepatitis B and C viruses cause motherto child infection. Transportation of such sample safely across different countries has always been a problem. To transport HIV samples safely from foreigncounties, we introduced the use of filter paper method
for blood and saliva. New primers were designed for
subtyping of HIV subtype B and E. We succeeded todetermine HIV from several countries. Now we aredeveloping primers for all subtypes. The PCR was useful for the samples from blood, saliva and breast milk.
Basic research of HIV infection was also done throughcollaboration with Prof. Muller in Germany; it was
found that exposure to gp120 of HIV induces apoptosis in neuronal cell culture. Moreover, I am still amember of National research group to find anti-HIV
candidate drugs. For the said purpose we screen antiHIV drugs distributed from pharmaceutical companies. We have found some effective chemicals in vitro.
Molecular Epidemiology of hepatitis B has been done
through the collaboration with Dr. Abe, NIH Japan.
Our results revealed that there was high prevalence of
HBV pre-S mutant in Asia, where there are highendemic regions of HBV, except for Japan. The
appearance of HBV mutation seemed to be correlated
to duration of persistent infection and genotypes of
HBV. In fact, we found higher prevalence of HBVmutation in hepatocellular carcinoma patients andgenotypes B and C. Mode of transmission of hepatitis
C infection in Japan is different to that in Asia. The
patients in most countries in Asia are due to co-infection with HIV but the majority of the patients inJapan are due to misuse of syringes.
Field studies
The field study of maternal and child health is also
integral part of our department. One project is “maternal and child health in minority groups in Asian countries”. Dr. Li Y, a researcher from Yunnan, China, hasconducted a project in rural minority area of Yunnan.
An interview of 2019 rural mother was conducted.
We found that maternal child-rearing behaviors differed significantly among ethnic groups as well as
between urban and rural areas. The behaviors weresignificantly associated with maternal education level,family type, family income, maternal age at delivery,
and number of children in the family. We continued
this project and found the malnutrition was especially
due to vitamin B1 deficiency. The main reason waseating too much polished rice, deficiency of vegetables
and longer time for cooking.

HIV and hepatitis viruses
HIV and hepatitis infections have become impor-345Current Research
Viruses
We have continued epidemiological studies of diarrheal viruses, HIV and hepatitis, especially in children. It will help achieve the final goal of decreasingmortality and morbidity caused by these diseases.
Enzyme immunoassay, (RT-) PCR, sequence analysis
and other methods are used for the diagnosis.
Immunochromatographic method is useful for diagnosis in outpatient clinics because of simplicity andrapidity.
We are trying to develop;(1) Diarrheal virus detection kit for rotavirus, entericadenovirus, astrovirus and Norwalk virus,(2) Hepatitis virus detection kit for hepatitis A, B, C
and E, and TTV.
To make antibodies for immunochromatograph,
we culture the viruses that grow in vitro, make synthetic peptides and make virus like particles withgenetic engineering. Rapid detection kit using gene
amplification is also proceeding.
Field research
Asia: As further activity in Yunnan, we conductedtraining course of maternal and child health for about
60 health workers and physicians who belong tominority areas. Education session focused on proper
cooking methodology, general physical examination,knowledge about HIV and other diseases. We madepamphlets and CD. Then, the health workers andphysicians teach the caregivers by using them.
Vitamin B1 is used in the area as a treatment. In addition, we used rapid methods of detecting viruses in
the areas.
A student from Vietnam, who worked before at
NGO GROUP, is studying in minority group in rural
mountain areas in north part of Vietnam. Physicalconditions of infants and attitudes of caregivers are
examined every 3 months after birth for 2 years. In
2003, a student will come from Laos. She will also
conduct similar research in Laos. Maternal and childhealth in minority groups in the border areas in
China, Thailand, Laos and Vietnam are examined andintervention is planned.
Japan: We are conducting a project of maternaland child health in ethnic minority in Japan. This
project started in 2001. Due to globalization, childrenof ethnic minority group have increased in Japan in
the last 10 years. However health check system, medical care and health insurance system are not enoughfor them. In addition, education of mother and child,communication among health workers and residents
are sometimes not sufficient for them. We have multipronged approach for the research. It includes (a)Ask

346

3000 local health workers about their activities onmother and child health in ethnic minority groups,(b)Interview health workers, physicians, NGO members and administrators deeply at two local areas how
they succeeded in high quality of health care forminority groups. (c)Some families stay in Japan over
the time than officially allowed. They do not receive
enough medical care. To improve the situation, we
examine it and make proposals for their better living.
(d)Lack of communication among themselves andwith health workers causes anxiety among them.
Proper and efficient communication system andhealth service system will be suggested. (e)In addition
to health care, education of mother and child is alsoneeded. Both are related and necessary for their better
health. All minorities in high class, middle class andlower class should be improved irrespective of ethnicity, religion or country of origin.
OthersOther activities are concurrently carried out in mydepartment are such as relation between developmentof voice / sucking and growth, mother’s
relaxation/infant development and touch care, maternal mortality and obesity in developed and developing countries. Moreover, other staffs in the division
have their own projects. We regularly communicate,
help and discuss together on our research agendas.

Future Prospects
21 st century has started in the wake of social
chaos. World has become a global village and emerging and reemerging infectious diseases are gaining
importance. While, exchanges of people and information are rapidly increasing via various modes of transportation and internet service. Under these circumstances, the health of mother, child, and elderly isespecially important. Our research of health in children among ethnic minority groups will become highly valuable. The results will contribute in making evidence based decision as regards maternal and childhealth.
In order to keep environment safe, a decrease ininfection diseases and malnutrition are equally important. Our diagnostic methods are useful to for theabove said cause and thus help fight against diseases.
Further development of vaccine, medicine and molecular epidemiological survey of infection is the need of
the time, to make the future safe for our generationsin the times to come.
Followings are schema of our research of past,present and future.

Research Grants
a: Health-welfare and labor science research(1) Maternal and child heath in multi-racial culturalsociety, a project of “general research on child andfamily” (2001-2004) 11,000,000 yen/3years(2) Research of survey of microorganism contamination in foods and evaluation of safety, a project of“general research on safety of food and chemicals”(2001-2004) 1,000,000 yen/year
b: Basic research supported by the ministry ofeducation and science(1) Diagnosis, molecular epidemiology, pathology, prevention and treatment of diarrheal viruses. (20022005) 18,891,000 yen(2002)(2) Molecular epidemiological study of maternal andchild infection in Asia. (2001-2004) 4,200,000
yen(2002)
c: Others(1) Research for gravity and infection supported by
space development project. 520,000 yen(2002)(2) Research for Nano-technology supported byhuman science foundation. Other small grants are
not shown. 7,500,000 yen/year

Select Publications
1. Ushijima H, Nishio O, Klocking R, Schroder HC,
Perovic S, Muller WEG: Exposure to gp120 of HIV-1
induces an increase release of aranchidonic acid in
rat neuronal cells followed by NMDA receptor-mediated neurotoxity. Eur J Neuroscience 7:13531359,1995.
2. Kakizawa J, Ushijima H, Ando S, Yamamoto Y, Oka
S, Ikeda Y, Muller WEG: Detection of HIV-1 RNA
and DNA in saliva by RT-PCR and PCR under different condition. Acta Paediatrica Japonica 38 : 218-223,
1996.

3. Yasui H, Kiyoshima J, Ushijima H. Passive protection
against rotavirus-induced diarrhea of pups born toand nursed by dams fed Bifidobacterium breve
YIT4064 J Infect Dis 172: 403-409, 199
4. Schroder HC, Ushijima H, Theis C, Seve A-P, HubertJ, Muller WEG. Expression of nuclear lectin carbohydrate-binding protein 35 in human immunodeficiency virus type 1-infected Molt-3 cells J AIDS 9:340348, 1995.
5. Saito K, Ushijima H, Nishio O, Oseto M, Motohiro H,
Ueda Y, Takagi M, Nakaya S, Ando T, Glass R,
Zaiman K. Detection of astroviruses from stool samples in Japan using reverse transcription and polymerase chain reaction amplification. Microbiology
and Immunology 39:825-828, 1995.
6. Wen L, Ushijima H, Kakizawa J, Fang Z-Y, Nishimura
S, Nishio O, Morokawa S, Motohiro T. Genetic variation in VP7 gene of human rotavirus serotype 2 (G2type) isolated in Japan, China, and Pakistan.
Microbiology and Immunology 39;911-915, 1995.
7. Ushijima H, Morikawa S, Mukoyama A, Nishio O.
Characterization of VP4 and VP7 of a murine
rotavirus (YR-1) isolated in Japan. Jpn. J. Med. Sci.
Biol. 48: 237-247, 1995.
8. Kakizawa J, Ushijima H, Morishita Y, Oka S, Ikeda Y,
Muller WEG. Diversity of HIV type 1 envelope V3loop region in saliva. Human retrovirus and AIDS,
12: 561-563, 1996.
9. Nakayama M, Ueda Y, Kawamoto H, Yue HJ, Saio K,
Nishio O, Ushijima H. Detection and sequencing of
Norwalk-like viruses from stool samples in Japan
using RT-PCR. Microbiology and Immunology 40:
317-320, 1996.
10. Yue Han-Jun, Hiroshi Ushijima. Detection and genotyping of astroviruses by RT-PCR and sequencing J.
Jap Assoc Infect Dis 70: 1220-1226, 1996
11. Perovic S, Schroder HC, Pergande G, Ushijima H,
Muller WEG. Downregulation of Bcl-2 and glutationelevel in neuronal cells in vitro treated with the prion
protein fragment (PrP106-126): Prevention by flupirtine Experimental Neurology 147:518-524, 1997.
12. Akihara S, Nakayama M, Kakizawa J, Bosu K, Izumi
K, Ushijima H. An outbreak of Norwalk-like virus347infection in Tokyo and Saitama in late 1995. J. Jap.
Assoc. Infect. Dis. 70: 840-841, 1996.
13. Wen L, Nakayama M, Yamanishi Y, Nishio O, Fang
Z-Y, Nakagomi O, Araki K, Nishimura S, Hasegawa
A, Muller WEG, Ushijima H. Genetic variation in the
VP7 gene of human rotavirus serotype 3 (G3 Type)isolated in China and Japan. Arch Virol 142:14811489,1997.
14. Kakizawa J, Ushijima H, Wen L, Ikeda Y, Oseto K.
Genetic analysis of the capsid region of human astrovirus serotype 3 isolated in Japan. Microbiol
Immunol 41: 637-640, 1977.
15. Schroder HC, Perovic S, Ushijima H, Muller WEG.
Mechanisms of prionｓｃ- and HIV-1 gp120 induced
neuronal cell death. Neuro Toxicology 19,683-688,
1998.
16. Kato H, Horino A, Sakurai S, Ushijima H, Komuro K,
Uchida T: Inhibition of tumor cell growth by murine
splenic adherent cells stimulated with IFN-ganma Int
Arch Allergy Immunol 115:115-120, 1998.
17. Hakamada-Taguchi R, Kato T, Ushijima H, Murakami
M, Uede T, Nariuchi H. Expression and co-stimulatory function of B7-2 on murine CD4+ T cells. Eur J
Immunol 28:865-873, 1998.
18. Matsui M, Ushijima H, Hachiya M, Kakizawa J,
Leying Wen, Oseto M, Morooka K, Kurz JB.
Detection of serotypes of astroviruses by reverse
transcription-polymerase chain reaction and homologies of the types by the sequencing of Japanese
Isolates. Microbiol. Immunol. 42: 539-547, 1998.
19. Cao X-R, Akihara S, Fang Z-Y, Nakagomi O, Ushijima
H. Genetic variation in the VP4 gene and the NSP4
gene of human rotavirus serotype 3 (G3 type) isolated in China and Japan. Microbiol Immunol 43(2):171175,1999.
20. Adhikary AK,Zhou Y, Kakizawa J, Numaga J, Akihara
S, Fang ZY, Ushijima H. Distribution of rotavirus VP4
genotype and VP7 serotype among Chinese Children.
Acta Paediatrica Japonica 40:641-643, 1998.
21. Abe T, Kobayashi M, Araki K, Ushijima H. Viral
infection-new concepts and treatment -infantile convulsion and viral infection- Brain and Development,
20:392-393, 1998.
22. Li Y, Liu J, Liu F, Guo G, Anme T, Ushijima H.
Maternal child-rearing behaviors correlates in rural
minority areas of Yunnan, China. Developmental andbehavioral pediatrics 21: 114-122, 2000
23. Li Y, Guo G, Shi A, Li Y, Anme T, Ushijima H.
Prevalence and correlates of Malnutrition among
Children in Rural Minority Areas of China. Pediatrics
International 41:549-556, 1999
24. Li Y, Kong L,Hotta M, Wongkhomthong S, Ushijima
H. Breat feeding in Bangkok, Thailand: the Current
Status, Maternal Knowledge, Attetude and Social
Support. Pediatrics International 41:648-654, 1999
25. Nishiwaki M, Yasumizu T, Hoshi K, Ushijima H. The
effect of pregnancy, lactation, and weaning on thebone mineral density of rats determined by dualenergy X-ray absorptionmetry. Endocrine Journal 46:
711-716, 1999.

348

26. Sakamoto T，Wang Q-H， Akihara S，KimＢ，
Nishimura S，Kaneshi K，Nakaya S，Ueda Y，
Sugita K，Motohiro T，Nishimura T，Ushijima H．
Molecular epidemiology of astrovirus in Japan from1995 to 1998 by RT-PCR with serotype-specificPrimers (1to 8) J Med Virol 61:326-331,2000.
27)Hachiya M, Matsui M, Sanogo M, Oseto M,
Morooka K, Ushijima H. Genetic variation in the capsid region of human astrovirus serotype 4 isolated inJapan. Microbiol Immunol 43(11),1067-1070,1999.
28. Ushijima H, Perovic S, Leuck J, Rytik P, Muller WEG,
HC Scroder.: Suppression of PrPsc- and HIV-1 gp120
induced neuronal cell death by sulfated colominic
acid. J Neurovirology 5:289-299, 1999.
29. Tanaka N,Shimoike T, Ishii K, Suzuki R, Suzuki T,
Ushijima H, Matsuura Y, Miyamura T. Selective
binding of hepatitis C virus core protein to synthetic
oligonucleotides corresponding to the 5’untranslated
region of the viral genome. Virology 270; 229-236,
2000.
30. Konishi K, Gu Y, Hatano I, Ushijima H. Effect of colominic acid on enteric-virus (rotavirus, poliovirus andCozsackievirus) infection in vitro. Japanese Journal of
Infectious Diseases. Jpn J Infect Dis 53:62-66, 2000.
31. Muller WEG, Laplanche JL, Ushijima H, Schroder
HC. Novel approaches in diagnosis and therapy of
Creutzfeldt-Jakob disease. Mechanisms of aging anddevelopment 31:116(2-3) 193-218, 2000 32 Ushijima
H. Rotavirus infection in Asia. Pediatrics
International. 42:392-394,2000
33. Kohno H, Akihara S, Nishio O, Ushijima H.
Development of a simple and rapid latex test for
rotavirus in stool. Pediatrics International 42:395400, 2000
34. Maneekarn N, Ushijima H. Epidemiology ofrotavirus infection in Thailand. Pediatrics
International 42:415-421, 2000
35. Nishio O, Matsui K, Lan DTP, Ushijima H, Isomura
S. Rotavirus infection among infants with diarrhea in
Vietnam. Pediatric International 42: 422-424, 2000
36. Zhou Y, Li L, Kim B, Kaneshi K, Nishimura S,
Kuroiwa T, Nishimura T, Sugita K, Ueda Y, Nakaya
T, Ushijima H. Rotavirus infection in children in
Japan. Pediatric International. 42:428-439, 2000
37. Gu Y, Gu Q, Kodama H, Muller WEG, Ushijima H.
Development of anti-rotavirus agents in Asia.
Pediatric International 42:440-447, 2000.
38. Wang QH, Kakizawa J, Wen L, Simizu M, Nishio O,
Fang Z, Ushijima H. Genetic Analysis of the Capsid
Region of Astroviruses. J Med Virol 64:245-255,2001.
39. Jonassen CM, Jonassen TO, Saif YM, Snodgrass DR,
Ushijima H, Shimizu M, Grinde B. Comparison ofcapsid sequences from human and animal astroviruses. J Gen Virol. 82:1061-1067, 2001.
40. Abe T, Kobayashi M, Araki K, Kodama H, Fujita Y,
Shinozaki T, Ushijima H. Infantile convulsions withmild gastroenteritis. Brain & Development 22:301306, 2000.
41. Adhikary AK, Numaga J, Kaburaki T, Kawashima H,
Kato S, Araike M, Miyata K, Simizu H, Suzuki E,

Yagyu F, Ushijima H. Rapid detection and typing of
oculopathogenic strain of subgenus D adenovirusesby fiber based PCR and restriction enzyme analysis.
Investigative ophthalmology & visual science. 42:
2010-2015, 2001.
42. Li Y, Shi A, Wan Y, Hotta M, Ushijima H. Childbehavior problems: Prevalence and correlates in rural
minority areas of China. Pediatrics International.
43:651-661, 2001.
43. Tani H, Nishimura M, Ushijima H, Miyamura T,
Matsuura Y. Characterization of cell-surface determinants important for baculovirus infection. Virology
279: 343-353, 2001.
44. Kudo S, Zhou Y, Cao XR, Yamanishi S, Nakaya S,
Ushijima H. Molecular characterization in the VP7,
VP4 and NSP4 genes of human rotavirus serotype 4(G4) isolated in Japan and Kenya. Microbiol Immunol
45 : 167-171, 2001.
45. Gu Y, Kodama H, Murata Y, Mochizuki D, Yanagawa
Y, Ushijima H, Shiba T, Lee CC. Mutations in the
ATP7A gene and copper metabolism in patients with
Menkes disease and occipital horn syndrome.
American Journal of Medical Genetics 99:217-222,
2001.
46. Kakizawa J, Nitta Y, Yamashita T, Ushijima H, Katow
S. Mutations of rubella virus vaccine TO-336 strain
occurred in the attenuation process of wild promotor
virus. Vaccine 19: 2793-2802, 2001.
47. Yamaguchi K, Ushijima H, Hisano M, Inoue Y,
Shimamura T, Hirano T, Muller WEG.
Immunomodulatory effect of gold sodium thiomalateof murine acquired immunodeficiency Syndrome.
Microbiol Immunol 45:549-555, 2001.
48. Zhou Y, Supawadee J, Khawan C, Tonusin S,
Peerakome S, Kim B, Kaneshi K, Ueda Y, Nakaya S,
Akatani K, Maneekarn N, Ushijima H.
Characterization of human rotavirus serotype G9 isolated in Japan and Thailand 1995 to 1997. J Med
Virol 65:619-628,2001.
49. Yagyu F, Ikeda Y, Sugiura W, Wongkhomthong SA,
Masuda M, Ushijima H. Differentiation of serotypes
B and E of human immunodeficiency virus type 1 bypolymerase chain reaction using novel env geneprimers. J Virol Methods;10: 11-20. 2002
50. Li Luibai, Thi Phuong Lan D, Hoa NT. Prevalence ofbreast-feeding and its correlates in Ho Chi Minh
City, Vietnum. Pediatrics International 44: 4754,2002. Other 20 English papers from 1995 to 2002.
(Total 70 papers)
10 majors are No.1, No.3, No.22, No.26, No..28, No.32,
No.38, No.40, No.41, No.48 and No.49.

349

Department of
Human Ecology
Outline and Research Objectives
In 1965, our department was established in the School of Health Sciences. From 1966 to
1972, however, there was no full-time professor in our department and thus its scientificactivities were limited. Between 1972 and 1976, Professor Akira Koizumi took the office andmade efforts for founding the bases in this new field, focusing on environmental health andpopulation analysis. For three years from 1976 when Professor Koizumi transferred to
Department of Public Health in our university, however, there was no professor again. In
1979, Professor Tsuguyoshi Suzuki transferred from Tohoku University, and since thenresearch activities of our department have flourished; in this period, particularly in the later
half, I fully assisted Professor Suzuki as an associate professor. In 1992, when I took hisplace, our department was reorganized as one of six departments in the newly established
School of International Health.
Regarding the research objectives, human ecology is unique in elucidating health andsurvival of humans at the individual and population levels, particularly from the relations orinteractions with their environment (not only the natural components but also the sociocultural components). Thus, the research of human ecology aims to integrate various aspects,
i.e. biomedical, environmental and sociocultural, as far as possible.
Since 1972 and particularly since 1979, the staff and students in our department haveconducted fieldworks and experimental works, with special interest in the linkage between
them. For the former, various populations in the Asia-Oceania region have been investigated, focusing on food-producing and consuming activities, nutritional and health status, population dynamics including births, deaths and migrations, and infectious and chronic degenerative (lifestyle-related) diseases; my own major contributions were related to the fieldworks. For the latter, toxicological and nutritional examinations have been conducted forbiological specimens (i.e. urine, blood and hair) and environmental samples (i.e. water andfoods), most of which were collected in our fieldworks; in addition, the associated animal
experiments have also been carried out.

Faculties and Students (in October 2002)
Professor and Chair Ryutaro Ohtsuka, D.Sc., since 1992.
Associate Professor Chiho Watanabe, Ph.D.
Associates ...............................2
Postdoctoral Fellow ................1 (foreigner).
Graduate Students ................13 (including 3 foreigners).
Research Students...................5 (including 3 foreigners).
Secretaries ...............................3 (1 full-time and 2 parttime).

Past Research and Major Accomplishments
It is reasonable to review the whole research activities of our department from the 1980s to the recent
past since I, as an associate professor from 1981 to
1992, actively participated in most of them, while
assisting, and collaborating with, the then Professor
T. Suzuki. For convenience, the major research fociare shown in Table 1, separately for the 1st period (up350

to 1992, when I became a professor) and the 2nd period (up to the late 1990s); in the same table, those in
the current and future periods are also mentioned. It
is noted that this table does not include the common
themes, such as nutritional adaptation, behavioral
adaptation and population dynamics, which havealways been pursued.
The most important goal of my research (withProfessor T. Suzuki and other staff) in the 1980s wasto establish the research framework of human ecology, focusing on human population ecology. Briefly, thesignificance of human population ecology or humanpopulation biology, which aims to elucidate humanadaptation at the population level not only in the
short term, e.g. within a year, but also in the longterm, e.g. through generations, had long been pointed
out by many human ecologists and human biologists
worldwide, but its systematic studies had been
lacked. For this purpose, my efforts were devoted tothe collaborative research in the Gidra-speaking popu-Table 1. The major research items, broken down into four periods
1st
2nd
3rd
4th
(-1992) (1990s) (current) (future)
Establishment of research framework++

Developments/Innovations of methodologies*
Human activity+
Nutritional/health status+++
Energy expenditure+++
Microdemography++
Specific themes**
Infectious diseases
Toxic elements
Lifestyle-related diseases
Harmonized development

*
**

+

++
+
+

+
+
+

(?)
(?)
(?)
(?)

+
++
+
+

+
+
++
++

Including only fieldwork-based methodologies.
The common themes are not included.
Infectious diseases: mostly, malaria in the earlier stages
and schistosomiasis in the later stages.
Toxic elements: mostly, mercury in the 2nd period andarsenic in the 3rd and 4th periods.
Lifestyle-related diseases: mostly, obesity, hypertension
and cardiovascular disorders.

lation in Papua New Guinea. The Gidra people aresuited to this purpose since they have maintained thebasic characteristics of a population, forming a unit ofmarriages and thus a unit of gene pool, despite thesmall number, i.e. about 2,000 in total, but theirmicroenvironmental conditions differ from village tovillage, e.g. inland, riverine and coastal terrains fromthe natural settings and in the degree of modernization from the sociocultural settings.
Our Gidra studies were characterized by the following two points. First, long-term field investigations were repeatedly conducted, based on observation, measurements and interviews, for the people’s
daily activity pattern, especially food procurements
and intakes, nutritional and health status and population dynamics. Second, biological specimens (blood,urine and hair) and environmental samples (foods andwater) were systematically collected for laboratory
analyses.
In tandem with the establishment of the researchframework, my efforts were directed to the developments and innovations of various biological, behavioral and microdemographic methods applicable tofield investigations. The major achievements included
1) genealogical-demographic analysis which has been
useful for reconstruction of long-term change in fertility, mortality and increase rates, 2) heart rate-monitoring method for estimation of energy expenditure
which has now become popular in human ecologicalfieldworks and 3) determination of sodium, potassium and urea nitrogen concentrations from urine samples, using filter papers, which has also been widely
used.
Following the findings from the Gidra studies,most of which were based on the combined efforts offieldwork and laboratory analysis, more than 40 original papers were published in international journals. In
addition, I, with Professor T. Suzuki, collected theimportant papers to edit a book in English, titled
Population Ecology of Human Survival: BioecologicalStudies of the Gidra in Papua New Guinea (No. 15 inSelect Publications), which has been distributed
worldwide to not only human ecologists but also scientists in the related fields. The 20 chapters of thisbook are categorized into four parts: “The ecology offood production,” “The ecology of food consumption,”“Nutrition and health” and “Population structure anddynamics.” Among various findings, some contributed
to the health promotion plans of the national andprovincial governments of Papua New Guinea: for
instance, disadvantageous effects of dietary modernization due to reduced intakes of essential micronutrients (e.g. zinc and potassium) and less effects of ironsupplementation to the anemia patients in malaria
endemic areas because their anemic states werecaused not by inadequate iron intake but by malariainduced haemolysis and dyserythropoiesis.
Since the late 1980s, I have increased the study
populations for more generalized understanding of
the people’s health and survival, based on the
research framework which was established. For
instance, our studies revealed an extraordinarily highmercury levels (in hair) among the Lake Murry population under artificially nonpolluted environment due
to large consumed amounts of fish in which mercury
was accumulated through biological concentration.
For another instance, our studies in the most modernized rural population, called Balopa, disclosed that
their prevalence rates of obesity and hypertension
were higher than those in many other populations
even in developed countries, mostly due to their lessenergy-consuming activity pattern and large amounts
of intake of fat-rich purchased foods.
Aside from the studies in Papua New Guinea, bothfield research and toxicological and nutritional experiments were conducted for various Japanese popula-351

tions, though only a few of them are listed in the
Select Publications.

Current Research
Since the late 1990s, my research foci have extended to clarify the effects of modernization on humanhealth and survival on the one hand, and on theother, the effects of some specific harmful environmental factors on human health and their mitigations.
In Papua New Guinea, our major efforts have been
devoted to the comparative analysis of rural (homeland) dwellers and rural-urban migrants of the same
populations, particularly the Huli and the Balopa,
whose genetic backgrounds markedly differ. For thispurpose, we have conducted research in both environments, the homelands and Port Moresby, i.e. the capital city, focusing on energy and nutrient intakes, energy expenditure and the prevalence of obesity, hypertension and other cardiovascular diseases and havedisclosed the significance of lifestyle change on them;protein status has been improved while obesity,
hypertension and many other cardiovascular risks
have been worsened. Furthermore, the comparison
between the two rural-urban migrant populations inPort Moresby revealed difference in the trigger factor,
obesity for the Balopa (an Austronesian group) andhigh lipoprotein-a level for the Huli (a NonAustronesian group).
Concurrently, I began research projects in several
Asian countries. Of them, the study for Bangladeshi
populations under arsenic hazards has been the major
target in the last several years. Despite that 35-50 million Bangladeshi people are recognized as a risk population of arsenic poisoning, systematic analyses, thedose-response relationships in particular, have seldom
been conducted. Our research contributed to concrete
elucidation for the interrelations among arsenic concentration of drinking water, urinary arsenic concentration and the extents of skin lesions (i.e. keratosis
and melanosis as the major symptoms of arsenic poisoning), in collaboration with dermatologists in our
University Hospital. Furthermore, in this research
project headed by me, many colleagues of different
disciplines, such as water chemists and public healthengineers from Switzerland and Japan and hydrologists from USA, have collaborated to seek integrated
ways for mitigation of arsenic hazards and sustainable
water supply systems in arsenic affected areas.
Energetics, i.e. the balance between energy intake
and energy expenditure, has been one of the important research themes in our department. The recent
studies have clarified its particularly significant roles
on degradation of health status in modernization
process among populations in Papua New Guinea,
Thailand and Bangladesh.

352

Finally, I began a biosocial research, which seeks
harmonized ways of community development andenvironmental conservation in the Asia-Oceania
region. The major reason why I organized this projectcame from my judgment that the basic idea of humanecology is useful for this issue. In this project, I
recruited not only biomedical scientists but rathersociocultural scientists from Japan and some other
Asian and Oceanian countries. At the beginning stage,
our findings have still been limited mostly due to difficulties in data collection by means of sociocultural
methods, but I will continue this research in the
future.

Future Prospects
My term of professor is not long, and thus I willconcentrate in the three research projects: the arsenic
hazards in Asian countries (not only Bangladesh but
several countries) and the harmonized community
development with environmental conservation in the
Asia-Oceania region, both which have already commenced, and a new project which has begun in theyear 2002 and will continue for three years. This new
project aims to elucidate the causes of markedly different prevalence of lifestyle-related diseases among
Asian, Melanesian and Polynesian populations. Basedon my previous research findings and in collaboration
with the staff in our school, particularly those from
Department of Human Genetics (headed by Professor
K. Tokunaga), the effects of both genetic factors andenvironmental (mostly lifestyle) factors will be simultaneously clarified.
My intentions for these three projects, with special
interest in collaborations with scientists of other disciplines, are as follows. The arsenic project, which has
been and will be sponsored by the Alliance for Global
Sustainability (the joint programs of Massachusetts
Institute of Technology in USA, Swiss Federal
University of Technology and our university), is
expected to contribute our human ecology research tothe integrated efforts for solution/mitigation of thisenvironmental issue, recognized very serious forhuman health and survival in several Asian countries.
The genetico-ecological project for lifestyle-related diseases aims at development of collaborative works
with many colleagues, particularly, those from
Departments of Human Genetics, Biomedical
Chemistry and Developmental Medical Sciences (all ofwhich belong to a large Department called
International Biomedical Sciences, together with ourdepartment). The harmonized development project is
expected to broaden human ecology aspects throughcollaboration with scientists of social medicine andsocial ecology in our school, i.e. a large Department ofInternational Social Medicine) and some others.

Research Grants
1. (as the Cluster Sub-leader, with many colleagues
from many countries) “People, land management andenvironmental change” organized by the United
Nations University and sponsored by the Global
Environmental Facility through the United Nations
Environmental Programme: 1998-99; 7,816,860 Yen(in total, for my own team).
2. (as the Leader, with 15 colleagues from Japan, USA,
Switzerland, Bangladesh and Nepal) “Mitigation ofgroundwater-derived arsenic hazards and sustainable
water supply system in Asian countries” sponsoredby the Alliance for Global Sustainability Programs;
1999-2002; 29,420,518 Yen (in total).
3. (as the Leader, with 30 colleagues from Japan, China,
Solomon Islands and Nepal) “Influences of development on local communities and their mitigations”
sponsored by the Japan Society for Promotion of
Science: 1999-2002; 184,000,000 Yen (in total).
4. (as the Leader, with 15 colleagues from Japan,
Solomon Islands and Indonesia) “Genetico-ecologicalstudies for high risks of lifestyle-related diseases in
Oceanian populations” sponsored by the
Monbukagaku-sho, Japan (Grant-in-Aid for Scientific
Research); 2002- 2004; 29,300,000 Yen (in total).
5. (as the Leader, with 10 colleagues from Japan, Papua
New Guinea, Malaysia and Nepal) “Application ofhigh-quality satellite images to analyses of land use
and environmental change” sponsored by the
Monbukagaku-sho, Japan (Grant-in-Aid for Scientific
Research); 2002-2005; 15,800,000 Yen (in total).

Select Publications
1. Ohtsuka, R. Oriomo Papuans: Ecology of Sago-Eatersin Lowland Papua. University of Tokyo Press, Tokyo,
1983.
2. Ohtsuka R, Kawabe T, Inaoka T, Suzuki T, Hongo T,
Akimichi T, Sugahara T. Composition of local andpurchased foods consumed by the Gidra in lowland
Papua. Ecology of Food and Nutrition, 15, 159-169,
1984.
3. Ohtsuka R, Kawabe T, Inaoka T, Akimichi T, Suzuki
T. Inter- and intra-population migration of the Gidrain lowland Papua. Human Biology, 57, 33-45, 1985.
4. Ohtsuka R, Inaoka T, Kawabe T, Suzuki T, Hongo T,
Akimichi T. Diversity and change of food consumption and nutrient intake among the Gidra in lowland
Papua. Ecology of Food and Nutrition, 16, 339-350,
1985.
5. Kawabe T, Ohtsuka R, Inaoka T, Akimichi T, Suzuki
T. Visual acuity of the Gidra in lowland Papua New
Guinea. Journal of Biosocial Science, 17, 361-369,
1985.
6. Ohtsuka R, Hongo T, Kawabe T, Suzuki T, Inaoka T,
Akimichi T, Sasano H. Mineral content of drinking
water in lowland Papua. Environment International,
11, 505-508, 1985.

7. Ohtsuka R. Low rate of population increase of theGidra Papuans in the past: A genealogical-demographic analysis. American Journal of Physical
Anthropology, 71, 13-23, 1986.
8. Inaoka T, Suzuki T, Ohtsuka R, Kawabe T, Akimichi
T, Takemori K, Sasaki N. Salt consumption, body fatness and blood pressure of the Gidra in lowland
Papua. Ecology of Food and Nutrition, 20, 55-66,
1987.
9. Suzuki T, Ohtsuka R. [Eds.] Human Ecology ofHealth and Survival in Asia and the South Pacific.
University of Tokyo Press, Tokyo, 1987.
10. Ohtsuka R, Suzuki T, Morita M. Sodium-rich tree ash
as a native salt source in lowland Papua. Economic
Botany, 41, 55-59, 1987.
11. Suzuki T, Watanabe S, Hongo T, Kawabe T, Inaoka
T, Ohtsuka R, Akimichi T. Mercury in scalp hair ofPapuans in the Fly estuary, Papua New Guinea.
Asia-Pacific Journal of Public Health, 2, 39-47, 1988.
12. Ohtsuka, R. Hunting activity and aging among theGidra Papuans: A biobehavioral analysis. American
Journal of Physical Anthropology, 79, 31-39, 1989.
13. Hongo T, Suzuki T, Ohtsuka R, Kawabe T, Inaoka T,
Akimichi T. Element intake of the Gidra in lowland
Papua: Inter-village variation and the comparison
with contemporary levels in developed countries.
Ecology of Food and Nutrition, 23, 293-309, 1989.
14. Hongo T, Suzuki T, Ohtsuka R, Kawabe T, Inaoka T,
Akimichi T. Hair element concentrations of theGidra in lowland Papua: The comparison withdietary element intakes and water element concentrations. Ecology of Food and Nutrition, 24, 167-179,
1990.
15. Ohtsuka R, Suzuki T. [Eds.] Population Ecology of
Human Survival: Bioecological Studies of the Gidra in
Papua New Guinea . University of Tokyo Press,
Tokyo, 1990.
16. Yoshinaga J, Minagawa M, Suzuki T, Ohtsuka R,
Kawabe T, Hongo T, Inaoka T, Akimichi T. Carbon
and nitrogen isotopic characterization for Papua New
Guinea foods. Ecology of Food and Nutrition, 26,
17-25, 1991.
17. Yoshinaga J, Suzuki T, Ohtsuka R, Kawabe T, Hongo
T, Imai H, Inaoka T, Akimichi T. Dietary selenium
intake of the Gidra, Papua New Guinea. Ecology ofFood and Nutrition, 26, 27-36, 1991.
18. Yoshinaga J, Suzuki T, Hongo T, Minagawa M,
Ohtsuka R, Kawabe T, Inaoka T, Akimichi T.
Mercury concentration correlates with the nitrogenstable isotope ratio in the animal food of Papuans.
Ecotoxicology and Environmental Safety, 24, 37-45,
1992.
19. Hongo T, Ohtsuka R, Nakazawa M, Kawabe T,
Inaoka T, Akimichi T, Suzuki T. Serum mineral andtrace element concentrations in the Gidra of lowland
Papua New Guinea: Inter-village variation and comparison with the levels in developed countries.
Ecology of Food and Nutrition, 29, 307-318, 1993.
20. Hongo T, Abe T, Ohtsuka R, Komai M, Okiyama T,
Amano K, Toyo-oka T, Suzuki T. Urinary mercury

353

monitoring of university staff and students occasionally exposed to mercury vapor. Industrial Health, 32,
17-27, 1994.
21. Ohtsuka R. Subsistence ecology and carrying capacity in two Papua New Guinea populations. Journal of
Biosocial Science, 26, 395-407, 1994.
22. Nakazawa M, Ohtsuka R, Kawabe T, Hongo T,
Suzuki T, Inaoka T, Akimichi T, Kano S, Suzuki M.
Differential malaria prevalence among villages of theGidra in lowland Papua New Guinea. Tropical andGeographical Medicine, 46, 350-354, 1994.
23. Hongo T, Ohtsuka R, Inaoka T, Kawabe T, Akimichi
T, Kuchikura Y, Suda K, Tohyama C, Suzuki T.
Health status comparison by urynalysis (dipsticktest) among four populations in Papua New Guinea.
Asia-Pacific Journal of Public Health , 7, 165-172,
1994.
24. Yoshida M, Ohtsuka R, Nakazawa M, Juji T,
Tokunaga K. HLA-DRB1 frequencies of
Non-Austronesian-speaking Gidra in south New
Guinea and their genetic affinities with Oceanian
populations. American Journal of Physical
Anthropology, 96, 177-181, 1995.
25. Ohtsuka R, Inaoka T, Umezaki M, Nakada N, Abe T.
Long-term subsistence adaptation to the diversified
Papua New Guinea environment: Human ecologicalassessments and prospects. Global Environmental
Change, 5, 347-353, 1995.
26. Abe T, Ohtsuka R, Hongo T, Suzuki T, Tohyama C,
Nakano A, Akagi H, Akimichi T. High hair and urinary mercury levels of fish eaters in the nonpolluted
environment of Papua New Guinea. Archives ofEnvironmental Health, 50, 367-373, 1995.
27. Yoshinaga J, Minagawa M, Suzuki T, Ohtsuka R,
Kawabe T, Inaoka T, Akimichi T. Stable carbon andnitrogen isotopic composition of diet and hair of
Gidra-speaking Papuans. American Journal of
Physical Anthropology, 100, 23-34, 1996.
28. Dejima Y, Fukuda S, Ichijoh Y, Takasaka K, Ohtsuka
R. Cold-induced salt intake in mice and catecholamine, renin and thermogenesis mechanisms.
Appetite, 26, 203-220, 1996.
29. Nakazawa M, Ohtsuka R, Kawabe T, Hongo T,
Inaoka T, Akimichi T, Suzuki T. Iron nutrition andanaemia in a malaria-endemic environment:
Haematological investigation of the Gidra-speaking
population in lowland Papua New Guinea. British
Journal of Nutrition, 76, 333-346, 1996.
30. Inaoka T, Ohtsuka R, Kawabe T, Hongo T, Suzuki T.
Emergence of degenerative diseases in Papua New
Guinea islanders. Environmental Sciences, 4, 79-93,
1996.
31. Nakazawa M, Ohtsuka R. Analysis of completed parity using microsimulation modeling. Mathematical
Population Studies, 6, 173-186, 1997.
32. Umezaki M, Ohtsuka R. Impact of rural-urban migration on fertility: A population ecology analysis in the
Kombio, Papua New Guinea. Journal of Biosocial
Science, 30, 411-422, 1998.

354

33. Ohtsuka R, Abe T, Umezaki M. Environmentally
Sound Agricultural Development in Rural Societies:
A Comparative View from Papua New Guinea andSouth China. UNESCO, Paris, 1998.
34. Sudo N, Ohtsuka R. Sleep patterns and sleep disorders among female shift workers in a computer factory of Japan. Journal of Human Ergology, 28, 39-47,
1999.
35. Umezaki M, Ishimaru H, Ohtsuka R. Daily timebudgets of long-distance commuting workers inTokyo Megalopolis. Journal of Biosocial Science, 31,
71-78, 1999.
36. Murayama N, Ohtsuka R. Seasonal fluctuation inenergy balance among farmers in Northeast
Thailand: The lack of response of energy intake tothe change of energy expenditure. European Journalof Clinical Nutrition, 53, 39-49, 1999.
37. Umezaki M, Yamauchi T, Ohtsuka R. Diet among the
Huli in Papua New Guinea Highlands when they
were influenced by the extended rainy period.
Ecology of Food and Nutrition, 37, 409-427, 1999.
38. Murayama N, Ohtsuka R. Heart rate indicators forassessing physical activity level in the field.
American Journal of Human Biology, 11, 647-657,
1999.
39. Natsuhara K, Inaoka T, Umezaki M, Yamauchi T,
Hongo T, Nagano M, Ohtsuka R. Cardiovascular risk
factors of migrants in Port Moresby from the highlands and island villages, Papua New Guinea.
American Journal of Human Biology, 12, 655-664,
2000.
40. Nakazawa M, Ohtsuka R, Hongo T, Inaoka T,
Kawabe T, Suzuki T. Serum biochemical data of theGidra in lowland Papua New Guinea: Considerationof their normal ranges. Journal of Nutrition andEnvironmental Medicine, 10, 153-162, 2000.
41. Ohashi J, Yoshida M, Ohtsuka R, Nakazawa M, Juji
T, Tokunaga K. Analysis of HLA-DRB1 polymorphism in the Gidra of Papua New Guinea. Human
Biology, 72, 337-347, 2000.
42. Yamauchi T, Ohtsuka R. Basal metabolic rate andenergy costs at rest and during exercise in rural- andurban-dwelling Papua New Guinea Highlanders.
European Journal of Clinical Nutrition, 54, 494-499,
2000.
43. Yamauchi T, Umezaki M, Ohtsuka R. Energy expenditure, physical exertion and time allocation among
Huli-speaking people in the Papua New Guinea
Highlands. Annals of Human Biology, 27, 571-585,
2000.
44. Umezaki M, Kuchikura Y, Yamauchi T, Ohtsuka R.
Impact of population pressure on food production:
An analysis of land use change and subsistence pattern in the Tari basin in Papua New Guinea
Highlands. Human Ecology, 28, 359-381, 2000.
45. Yamauchi T, Umezaki M, Ohtsuka R. Influence ofurbanisation on physical activity and dietary changes
in Huli-speaking population: A comparative study ofvillage dwellers and migrants in urban settlements.
British Journal of Nutrition, 85, 65-73, 2001.

46. Yamauchi T, Umezaki M, Ohtsuka R. Physical activity and subsistence pattern of the Huli, a Papua New
Guinea Highland population. American Journal of
Physical Anthropology, 114, 258-268, 2001.
47. Umezaki M, Natsuhara K, Ohtsuka R. Protein content and amino acid scores of sweet potatoes in
Papua New Guinea Highlands. Ecology of Food andNutrition, 40, 471-480, 2001.
48. Watanabe C, Inaoka T, Kadono T, Nagano M,
Nakamura S, Ushijima K, Murayama N, Miyazaki K,
Ohtsuka R. Males in rural Bangladeshi communities
are more susceptible to chronic arsenic poisoning
than females: Analyses based on urinary arsenic.
Environmental Health Perspectives, 109, 1265-1270,
2001.
49. Sudo N, Ohtsuka R. Nutrient intake among female
shift workers in a computer factory in Japan.
International Journal of Food Sciences and Nutrition,
52, 367-378, 2001.
50. Umezaki M, Yamauchi T, Ohtsuka R. Time allocation
to subsistence activities among the Huli in rural andurban Papua New Guinea. Journal of Biosocial
Science, 34, 133-137, 2002.

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Department of
Biomedical Chemistry
Outline and Research Objectives
Aim of our department is to contribute to global health and welfare from basic research.
Our department, formerly named Biochemistry and Nutrition was renamed on April 1st,
1996 to The Department of Biomedical Chemistry as newly affiliating with Biomedical
Science Division of International Health, Graduate School of Medicine, The University ofTokyo. Prof. Kita has moved from The Institute of Medical Science, The University of Tokyo
on March 1st, 1998. Therefore, we are quite new and all of our members are highly motivated.

Faculties and Students
Professor and Chair

KITA Kiyoshi, Dr. of Pharmacy(1998-)
Lecturer
WATANABE Yoh-ichi, Dr. ofTechnology
Associate .................................3
Postdoctoral Fellow ................2
Graduate student ..................12
Research student.....................4
Secretary .................................1

Past Research and Major Accomplishments
Energy metabolism is essential for the survival,continued growth and reproduction of living organisms. From the standpoint of biological adaptation,
we have been studying on the molecular mechanismof energy transducing systems such as mitochondrialand bacterial respiratory chain. Because energy metabolisms of pathogenic organisms such as bacteria andparasites are quite different from that of mammalianhost and these unique aspects of pathogens would bepromising targets for chemotherapy.
For the first step to understand a mechanism ofadaptation to the change of oxygen availability, composition and its change during aerobic growth ofEscherichia coli was studied. Finally, we found two
different terminal oxidases, cytochrome bo and bd
complexes. Cytochrome bo complex is expressed inearly log phase and possesses proton-pumping activity, while cytochrome bd complex is expressed fromlate log phase to stationary phase with higher affinity
to oxygen than cytochrome bo complex. Thus, wefound that E. coli keep their energy supply by changing the respiratory chain during their growth.
Then, we have started our research on the biological strategy to adapt to different oxygen availability ineukaryotes. Purpose of this project is multiple. We

356

would like to know the common strategy to maintainenergy supply in different environments and also tounderstand unique feature of energy metabolism inpathogenic organisms. If we could understand themechanism clearly, it would be quite important information to fight against infectious diseases caused bybacteria and parasites in addition to the basic knowledge on the molecular mechanism of biological adaptation. As a model system, we selected Ascaris suum
which is a parasitic nematode resides in our small
intestine where oxygen tension is quite limited,
because 1) they are rather large in size allowing biochemical study, 2) their life cycle has been well characterized and 3) they are closely related to
Caenorhabditis elegans which is well known free-living nematode, of which whole genome sequence has
been reported.

Current Research
I. Human mitochondria
1) succinate dehydrogenase2) mitochondrial myopathy
Succinate-ubiqinone oxidoreductase (complex II
and/or SQR) is an important enzyme complex in boththe tricarboxylic acid cycle and the aerobic respiratory
chains of eukaryotic mitochondria and prokaryotic
cells. Complex II catalyzes the oxidation of succinate
to fumarate (succinate dehydrogenase: SDH) andtransfers its reducing equivalent to ubiquinone. To
elucidate the molecular basis of a mitochondrial disease caused by a deficiency in the SDH activity ofcomplex II, we cloned the cDNAs for all four subunitsof human liver complex II. During this study, we
found two isoforms of complex II containing Fp subunits of liver type and heart type.
Then we have mapped large (CybL) and small(CybS) subunits of cytochrome b in complex II ofhuman mitochondria to chromosome 1q21 and
11q23, respectively. Recently, the human SDHD geneencoding CybS was cloned and characterized. The
gene comprises four exons and three introns extending over 19 kb. Sequence analysis of the 5’ promoter
region showed several motifs for the binding of transcription factors including nuclear respiratory factors
NRF-1 and NRF-2 at positions -137 and -104, respectively. In addition to this gene, six pseudogenes of
CybS were isolated and mapped on the chromosome.
II. Ascaris suum and Caenorhabditis elegans
1) molecular mechanism of adaptation to low oxygen
tension (regulation of gene expression of mitochondrial proteins)2) mitochondrial quinol-fumarate reductase (structure-function relationship, enzyme evolution)3) C. elegans as a model system of parasitic nematodes and ageing (expression of foreign genes,
gene knockout, oxygen stress)
Parasites have developed a variety of physiological
functions necessary for existence within the specialized environment of the host. Regarding energy
metabolism, which is an essential factor for survival,
parasites adapt to low oxygen tension in host mammals using metabolic systems that are very different
from that of the host. The majority of parasites does
not use the oxygen available within the host, but
employ systems other than oxidative phosphorylation
for ATP synthesis.
In addition, all parasites have a life cycle. In manycases, the parasite employs aerobic metabolism during
their free-living stage outside the host. In such systems, parasite mitochondria play diverse roles. In particular, marked changes in the morphology and components of the mitochondria during the life cycle arevery interesting elements of biological processes suchas developmental control and environmental adaptation. Recent our research on adult A. suum has shownthat the stage-specific mitochondrial complex II plays
an important role in the anaerobic energy metabolismof parasites inhabiting hosts, by acting as quinolfumarate reductase (QFR). Sequence analysis of thesubunits in the enzyme revealed that mitochondrial
QFR is new enzyme evolved by “reverse evolution” of
SQR rather than direct evolution from bacterial QFR.
We have been studying free-living nematode, C.
elegans as a control nematode living in an aerobic
condition. In addition, we also have been studying
ageing by using C. elegans. Our study on long-lived
clk-1 mutants of C. elegans demonstrated that biosynthesis of UQ is dramatically altered in mutant mitochondria. Demethoxy ubiquinone (DMQ), that accumulates in clk-1 mutants in place of UQ, may contribute to the extension of life span. We proposed thepossible mechanisms of life span extension in clk-1
mutants, with particular emphasis on the electrochemical property of DMQ. On the other hand, in the
short-lived mev-1 mutant, a point of mutation glycine
71 of CybL in SQR results in hypersensitivity tooxidative stress. We demonstrated the production ofreactive oxygen species (ROS) was higher in the
mutant complex II than wild type, indicating ROS is
responsible for the short life span of mev-1 mutant.

III. Malaria and Trypanosome (Plasmodium falciparum, Trypanosoma brucei,Trypanosoma cruzi)
1) characterization of mitochondria as a target for thechemotherapy2) molecular biology of mitochondrial DNA3) structure based drug design (SBDD)
The mitochondria of parasitic protozoa that havebeen most studied are those of Trypanosoma brucei,
which is the causative agents of African trypanosomiasis in human and nagana in cattle. Parasitic protozoa
living within a mammalian host do not generally use
oxygen, and instead synthesize ATP via the glycolytic
pathway. In comparison to research on helminthes,
research on mitochondrial electron transfer of protozoa during the developmental stage within the host islacking. However, mitochondria have become a focusof chemotherapy, and researches, particularly on
mitochondria of the malaria parasite and trypanosomes are currently progressing.
To study the unique properties of Plasmodium
complex II, we have cloned and characterized thegenes for the flavoprotein subunit (Fp) and the ironsulfur protein subunit (Ip) subunits of complex II.
This is the first report of the primary structure of protozoan complex II. Interestingly, a P. falciparum-specific insertion and a unicellular organism-specific deletion were found in the amino acid sequence of Fp.
Despite the fact that Fp and Ip are generally well-conserved subunits, the structure of both subunits in themalaria parasite clearly differs considerably from thatof the host.
Cyanide-insensitive trypanosome alternative oxidase (TAO) is the terminal oxidase of the respiratory
chain of long slender bloodstream forms of theAfrican trypanosome. TAO is a cytochrome-independent, cyanide-insensitive quinol oxidase. These characteristics are distinct from those of the bacterial quinoloxidase, a protein that belongs to the heme-copper
terminal oxidase superfamily. TAO has been targeted
for the development of anti-trypanosomal drugs
because it does not exist in the host. We found themost potent inhibitor of TAO to date, ascofuranone, acompound isolated from phytopathogenic fungus,
Ascochyta visiae. The inability to purify stable TAO
has severely hampered biochemical studies of the357

alternative oxidase family. Recently, we were able topurify recombinant TAO to homogeneity from E. coli
membranes using the detergent digitonin. Kinetic
analysis of the purified TAO revealed that ascofuranone is a competitive inhibitor of ubiquinol oxidase
activity.
IV. Escherichia coli
1) succinate dehydrogenase complex (mechanism ofmolecular assembly, electron transport mechanismin the complex)2) regulation of energy supply

E. coli is one of the useful model systems to investigate biological reactions common to living organisms. In order to investigate the role of the heme in
the assembly of complex II, we used a hemH (encodes
ferrochelatase) mutant of E. coli lacking the ability toinsert iron into the porphyrin ring. The hemH mutant
failed to synthesize functional complex II in the cytoplasmic membrane and the catalytic portion of complex II, Fp and Ip, was localized in the cytoplasm of
the cell. In contrast, complex II was assembled in themembrane of a heme-permeable and hemH double
mutant when hemin was present in the culture. Onlya small amount of succinate-ubiquinone reductase(SQR) activity was found in the membrane whenhemin was replaced by non-iron metalloporphyrins;
Mn-, Co-, Ni-, Zn-, Cu-protoporphyrin IX or protoporphyrin IX. These results indicate the indispensabilityof iron of heme for the functional assembly of complex II in the cytoplasmic membrane of E. coli, andadd a new aspect to the biological role of heme in themolecular assembly of the multi-subunit enzyme complex.

Future Prospects
Dynamic rearrangement of the respiratory chain
during parasite life cycle that we found is a key element of their adaptation to different environments. In
the case of mammals, cells are able to sense decreased
oxygen and activate response systems, including
hypoxia-inducible factor-1 (HIF-1)–mediated transcriptional activation of several genes. Quite recently,
HIF-β and a homologue of HIF-1α were found in thefree-living nematode C. elegans. Although it is notknown whether such a pathway exists in the parasites, A. suum shows a very clear transition between
larval and adult metabolic systems. Thus, A. suum is
an excellent model system for studying the regulationof transcription by the oxygen level in the environment.
Current research on C. elegans including our result
has revealed that oxygen concentration, its availability, and oxidative stress can produce a variety of inter-358

esting phenotypes such as modified life spans. As
described above, in the long-lived C. elegans mutant
clk-1, UQ biosynthesis is altered so that mitochondria
do not possess detectable levels of UQ-9 but instead
contain the UQ biosynthesis intermediate DMQ-9. On
the other hand, in the short-lived mev-1 mutant, apoint of mutation glycine 71 of CybL in SQR results in
hypersensitivity to oxidative stress. These findings
indicate that the respiratory chain plays an importantrole in sensing and responding to the oxygen level inthe environment. Consistent with this, recent reportssuggest that complex II functions as an oxygen sensor
in human. We would like to understand a molecularmechanism of biological adaptation to the change ofoxygen availability by using these nematodes andhuman systems. Diversity of complex II was also
found in human complex II. Studies on the humanrespiratory chain will be achieved from the viewpointof physiological role of energy supply and related diseases.
It remains unclear which critical factor determines
the catalytic direction of electron transfer in complex
II. Biochemical and molecular biological studies
showed that both the properties and the primarystructures of mitochondrial and bacterial quinolfumarate reductase (QFR) differ significantly.
Identification of the amino acid residues of mitochondrial QFR responsible for the directional specificity of
its catalysis should help to clarify the molecular mechanism of this “new” parasite enzyme. Furthermore,investigations of the molecular pathways of parasite
survival may provide insight into general mechanismsof biological adaptation.
Finally, we would like to develop anti-parasitic
drugs which should be used in endemic areas. As
described in “Current Research”, the parasite metabolic system is an attractive target for chemotherapy
because it is different than the host metabolic systemand because it is essential for parasite survival. TAO
in trypanosome mitochondria and its specificinhibitor, ascofuranone are excellent examples in thisregard. As pointed out by Dr. Opperdoes, ascofuranone is the most potent TAO inhibitor to date.
Because of its high selectivity and low toxicity, ascofuranone is a potential candidate for the next generation of anti-trypanosomal agents. Indeed, current
studies are examining the molecular mechanism of
ascofuranone and ways of improving its efficacy invivo. We already have started a trial by using goat in
Africa. One problem that must be solved is that glycerol must be administered along with ascofuranone,
to inhibit the reverse reaction of glycerol kinase.
Although glycerol is not toxic, it is not practical to
give the large dose (3g /kg) required, and an inhibitorof the reverse reaction of glycerol kinase is expected
to be more useful. Fortunately, glycerol kinase hasbeen cloned and the recombinant enzyme can be overexpressed. We are now trying to screen and find a
specific inhibitor of glycerol kinase.
Thus, we would like to contribute to global healthand welfare from basic research and train young scientists to be thoughtful and powerful colleagues.

Research Grants
1. “Molecular and biochemical studies on Plasmodium
mitochondria” from a Grant-in-aid for scientificresearch on priority areas from the Ministry of
Education, Science, Culture and Sport, Japan
40,000,000 yen (2001-)
2. “Crystalization of parasite proteins and molecular
design of anti-parasitic drugs” from Pilot Applied
Research Project for the Industrial Use of Space” ofthe National Space Development Agency of Japan(NASDA) and Japan Space Utilization Promotion
Center (JSUP).
30,000,000 yen (2000-)
3. “New anti-trypanosome drug, ascofuranone” from a
Grant for research on emerging and re-emerging
infectious diseases from the Ministry of Health andWelfare120,000,000 yen (1999-2001)
4. “Molecular structure and role of mitochondrial
fumarate reductase from Ascaris suum ” from a
Grant-in-aid for scientific research on priority areasfrom the Ministry of Education, Science, Culture andSport, Japan
13,200,000 yen (1999-2000)
5. “Molecular properties of Plasmodium mitochondria
in the erythrocytic stages” from a Grant-in-aid for scientific research on priority areas from the Ministryof Education, Science, Culture and Sport, Japan
28,000,000 yen (1997-2000)

Select Publications
1. Kita, K., Yamato, I., and Anraku, Y. Purification andproperties of cytochrome b 556 in the respiratory
chain of aerobically grown Escherichia coli K12. J.
Biol. Chem. 253, 8910-8915, 1978.
2. Kita, K., Kasahara, M., and Anraku, Y. Formation ofa membrane potential by reconstituted liposomes
made with cytochrome b562-o complex, a terminal
oxidase of Escherichia coli K12. J. Biol. Chem. 257,7933-7935, 1982.
3. Kita, K., Konishi, K., and Anraku, Y. Terminal oxidases of Escherichia coli aerobic respiratory chain. I.
Purification and properties of cytochrome b 562- ocomplex from cells in the early exponential phase of
aerobic growth. J. Biol. Chem. 259, 3368-3374, 1984,
II. Kita, K., Konishi, K., and Anraku, Y. Purificationand properties of cytochrome b558-d complex from
cells grown with limited oxygen and evidence ofbranched electron carrying systems. J. Biol. Chem.
259, 3375-3381, 1984.

4. Kita, K., Takamiya, S., Furushima, R., Ma, Y., andOya, H. Complex II is a major component of the respiratory chain in the muscle mitochondria of Ascaris
suum with high fumarate reductase activity. Comp.
Biochem. Physiol. 89, 31-34, 1988.
5. Kita, K., Takamiya, S., Furushima, R., Ma, Y., Suzuki,
H., Ozawa, T., and Oya, H.Electron-transfer comlexof Ascaris suum muscle mitochondria. III.
Composition and fumarate reductase activity of complex II. Biochim. Biophys. Acta 935, 130-140, 1988.
6. Hata-Tanaka, A., Kita, K., Furushima, R., Oya, H.,
and Itoh, S. ESR studies on iron-sulfur clusters ofcomplex II in Ascaris suum mitochondria which
exhibits strong fumarate reductase activity. FEBS
Lett. 242, 183-186, 1988.
7. Kita, K., Vibat, C., Meinhardt, S., Guest, J., andGennis, R. One-step purification from Escherichia
coli of complex II (succinate-ubiquinone oxidoreductase) associated with succinate reducible cytochromeb556. J. Biol. Chem. 264, 2672-2677, 1989.
8. Kita, K., Oya, H., Gennis, R., Ackrell, B.A.C., andKasahara, M. Human complex II (succinateubiquinone oxidoreductase): cDNA cloning of ironsulfur (Ip) subunit of liver mitochondria. Biochem.
Biophys. Res. Commun. 166, 101-108, 1990.
9. Furushima, R., Takamiya, S., Okochi, I., Kita, K.,
Aoki, T., and Oya, H. A comparative study of theeffects of phebrol on the respiratory chains of rat
liver, Biomphalaria glabrata and Oncomelania
nosophora. Comp. Biochem. Physiol. 99C, 191-199,
1991.
10. Kita, K. Electron-transfer complexes of mitochondria
in Ascaris suum. Parasitol. Today 8, 155-159, 1992.
11. Hirawake, H., Wang, H., Kuramochi, T., Kojima, S.,
and Kita, K. Human Complex II (succinateubiquinone oxidoreductase): cDNA cloning of theflavoprotein (Fp) subunit of liver mitochondria. J.
Biochem. 116, 221-227, 1994.
12. Kuramochi, T., Hirawake, H., Kojima, S., Takamiya,S., Furushima, R., Aoki, T., Komuniecki, R., and Kita,
K. Sequence comparison between the flavoprotein
subunit of the fumarate reductase (Complex II) of theanaerobic parasitic nematode, Ascaris suum and thesuccinate dehydrogenase of the aerobic, free-living
nematode, Caenorhabditis elegans. Mol. Biochem.
Parasitol. 68, 177-187, 1994.
13. Watanabe, Y., Tsurui, H., Ueda, T., Furushima, R.,
Takamiya, S., Kita, K., Nishikawa, K., and Watanabe,
K. Primary and higher order structures of nematode(Ascaris suum) mitochondrial tRNAs lacking either
the T or D stem. J. Biol. Chem. 269, 22902-22906,
1994.
14. Saruta, F., Kuramochi, T., Nakamura, K., Takamiya,S., Yu, Y., Aoki, T., Sekimizu, K., Kojima, S., andKita, K. Stage-specific isoforms of ComplexII (succinate-ubiquinone oxidoreductase) in mitochondria
from the parasitic nematode, Ascaris suum. J. Biol.
Chem. 270, 928-932, 1995.
15. Kuramochi, T., Kita, K., Takamiya, S., Kojima, S., andHayasaki, M. Comparative study and cDNA cloning359

of the flavoprotein subunit of mitochondrial complex
II (succinate-ubiquinone oxidoreductase: fumarate
reductase) from the dog heartworm, Dirofilaria
immitis. Comp. Biochem. Physiol. 111B, 491-502,
1995.
16. Nakamura, K., Yamaki, M., Sarada, M., Nakayama,S., Vibat, C.R.T., Gennis, R.B., Nakayashiki, T.,
Inokuchi, H., Kojima, S., and Kita, K. Two hydrophobic subunits are essential for the heme b ligation andfunctional assembly of complex II (succinateubiquinone oxidoreductase) from Escherichia coli. J.
Biol. Chem. 271, 521-527, 1996.
17. Takamiya, S., Hirawake, H., Kuramochi, T., Aoki, T.,
Kojima, S., and Kita, K. cDNA cloning for mitochondrial cytochrome c and its adult-specific isoform
from Ascaris suum. Mol. Biochem. Parasitol. 76, 293297, 1996.
18. Ohtsuki, T., Kawai, G., Watanabe, Y., Kita, K.,
Nishikawa, K., and Watanabe, K. Preparation of biologically active Ascaris suum mitochondrial
tRNAMet with a TV-replacement loop by ligation ofchemically synthesized RNA fragments. Nucleic
Acids Res. 24, 662-667, 1996.
19. Noor Rain, A., Furuta, T., Radzan, T., Kita, K.,
Kojima, S., and Mak, J.W. Development of a new
diagnostic method for Plasmodium falciparum infection using a reverse transcriptase-polymerase chain
reaction. Am. J. Trop. Med. Hyg. 54, 162-163, 1996.
20. Saruta, F., Hirawake, H., Takamiya, S., Ma, Y-C.,
Aoki, T., Sekimizu, K., Kojima, S., and Kita, K.
Cloning of a cDNA encoding the small subunit ofcytochrome b558 (cybS) of mitochondrial fumarate
reductase (Complex II) from adult Ascaris suum .
Biochim. Biophys. Acta 1276, 1-5, 1996.
21. Minagawa, N., Yabu, Y., Kita, K., Ohta, N., Meguro,
K., Sakajo, S., and Yoshimoto,A. An antibiotic,
Ascofuranone, specifically inhibits respiration and invitro growth of long slender bloodstream forms of
Trypanosoma brucei brucei. Mol. Biochem. Parasitol.
81, 127-136, 1996.
22. Lim, P.K.C., Watanabe, Y., Tan, S.K., Furuta, T.,
Aoki, T., Kojima, S., Kita, K., Sharma, Y.D., and Mak,
J.W. Cytochrome c oxidase III (COIII) gene of
Plasmodium vivax: Complete coding sequence andits expression in the erythrocytic stage of the parasite. Jpn. J. Parasitol. 45, 373-383, 1997.
23. Nara, T., Tanabe, K., Mahakunkijcharoen, Y., Osada,Y., Matsumoto, N., Kita, K., and Kojima, S. The B cell
epitope of paramyosin recognized by a protective
monoclonal IgE antibody to Schistosoma japonicum.
Vaccine 15, 79-84, 1997.
24. Watanabe, Y., Tsurui, H., Ueda, T., Furushima, R.,
Takamiya, S., Kita, K., Nishikawa, K., and Watanabe,
K. Primary sequence of mitochondrial tRNAArg of anematode Ascaris suum: occurrence of unmodified
adenosine at the first position of the anticodon.
Biochim. Biophys. Acta 1350, 119-122, 1997.
25. Kita, K., Hirawake, H., and Takamiya, S.
Cytochromes in the respiratory chain of helminth
mitochondria. Int. J. Parasitol. 27, 617-630, 1997.

360

26. Hirawake, H., Kita, K., and Sharma, Y.D. Variation in
the C-terminal repeats of the knob-associated histidine-rich protein of Plasmodium falciparum .
Biochim. Biophys. Acta 1360, 105-108, 1997.
27. Isobe, K., Kishino, S., Inoue, K., Takai, D., Hirawake,
H., Kita, K., Miyabayashi, S., and Hayashi, J.
Identification of inheritance modes of mitochondrial
diseases by introduction of pure nuclei from mtDNAless HeLa cells to patient-derived fibroblasts. J. Biol.
Chem. 272, 12606-12610, 1997.
28. Watanabe, Y., Kita, K., Ueda, T., and Watanabe, K.
cDNA sequence of a translational elongation factor
Ts homologue from Caenorhabditis elegans: mitochondrial factor-specific features found in the nematode homologue peptide. Biochim. Biophys. Acta
1353, 7-12, 1997.
29. Hirawake, H., Taniwaki, M., Tamura, A., Kojima, S.,
and Kita, K. Cytochrome b in human complex II (succinate-ubiquinone oxidoreductase): cDNA cloning of
the components in liver mitochondria and chromosome assignment of the genes for the large (SDHC)and small (SDHD) subunits to 1q21 and 11q23.
Cytogenet. Cell Genet. 79, 132-138, 1997.
30. Vibat, C.R.T., Cecchini, G., Nakamura, K., Kita, K.,
and Gennis, R.B. Localization of histidine residues
responsible for heme axial ligation in cytochromeb556 of complex II (succinate:ubiquinone oxidoreductase) in Escherichia coli. Biochemistry 37, 4148-4159,
1998.
31. Fukai, Y., Amino, H., Hirawake, H., Yabu, Y., Ohta,
N., Minagawa, N., Sakajo, S., Yoshimoto, A., Nagai,
K., Takamiya, S., Kojima, S., and Kita, K. Functional
expression of the ascofuranone-sensitive
Trypanosoma brucei brucei alternative oxidase in thecytoplasmic membrane of Escherichia coli. Comp.
Biochem. Physiol. 124, 141-148, 1999.
32. Hirawake, H., Taniwaki, M., Tamura, A., Amino, H.,
Tomitsuka, A., and Kita, K. Characterization of thehuman SDHD gene encoding the small subunit ofcytochrome b (cybS) in mitochondrial succinateubiquinone oxidoreductase. Biochim. Biophys. Acta
1412, 295-300, 1999.
33. Amino, H., Wang, H., Hirawake, H., Saruta, F.,
Mizuchi, D., Mineki, R., Shindo, N., Murayama, K.,
Takamiya, S., Aoki, T., Kojima, S., and Kita, K. Stagespecific isoforms of Ascaris suum complex II: thefumarate reductase of the parasitic adult and the succinate dehydrogenase of free-living larvae share acommon iron-sulfur subunit. Mol. Biochem.
Parasitol. 106, 63-76, 2000.
34. Takeo, S., Kokaze, A., Ng, C. S., Mizuchi, D.,
Watanabe, J., Tanabe,K., Kojima, S., and Kita, K.
Succinate dehydrogenase in Plasmodium falciparummitochondria: Molecular characterization of the
SDHA and SDHB genes for the catalytic subunits,
the flavoprotein (Fp) and iron-sulfur (Ip) subunits.
Mol. Biochem. Parasitol. 107, 191-205, 2000.
35. Omura, S., Miyadera, H., Ui, H., Shiomi, K.,
Yamaguchi, Y., Masuma, R., Nagamitsu, T., Iwai, Y.,
Takano, D., Sunazuka, T., Harder, A., Klbl, H.,

Namikoshi, M., Miyoshi, H., Sakamoto, K., and Kita,
K. An anthelmintic compound nafuredin shows
selective inhibition of complex I in helminth mitochondria. Proc. Natl. Acad. Sci. USA, 98, 60-62, 2001.
36. Nihei, C., Nakayashiki, T., Nakamura, K., Inokuchi,
H., Gennis, R. B., Kojima, S., and Kita, K. Abortive
Assembly of Succinate-Ubiquinone Reductase(Complex II) in a Ferrochelatase-deficient Mutant ofEscherichia coli. Mol. Genet. Genomics. 265, 394404, 2001.
37. Miyadera, H., Amino, H., Hiraishi, A., Taka, H.,
Murayama, M., Miyoshi, H., Sakamoto, K., Ishii, N.,
Hekimi, S., Kita, K. Altered quinone biosynthesis in
the long-lived clk-1 mutants of Caenorhabditis elegans. J. Biol. Chem., 276, 7713-7716, 2001.
38. Ohtsuki, T., Watanabe, Y., Takemoto, C., Kawai, G.,
Ueda, T., Kita, K., Kojima, S., Kaziro, Y., Nyborg, J.,
and Watanabe, K. An “Elongated” translation elongation factor Tu for truncated tRNAs In nematodemitochondria. J. Biol. Chem. 276, 21571-21577, 2001.
39. Takashima, E., Takamiya, S., Takeo, S., Mi-ichi, F.,
Amino, H.and Kita, K. Isolation of mitochondria
from Plasmodium falciparum showing dihydroorotate dependent respiration. Parasitol. Int. 50, 273278, 2001.
40. Ishii, N., Kita, K. and Hartman, P. S. Mitochondrial
contributions to aging in the nematode
Caenorhabdiitis elegans. Current Genomics 2, 349355, 2001.
41. Matsuzaki , M., Kikuchi, T., Kita , K., Kojima, S., andKuroiwa, T. Large amount of apicoplast nucleoid
DNA and its segregation in Toxoplasma gondii .
Protoplasma, 218, 180-191, 2001.
42. Levavasseur, F., Miyadera, H., Sirois, J., Tremblay,
M., Kita, K., Shoubridge, E. and Hekimi, S.
Ubiquinone is necessary for mouse embryonic development but is not essential for mitochondrial respiration. J. Biol. Chem. 276, 46160-46164, 2001.
43. Kita, K., Hirawake, H., Miyadera, H., Amino, H., andTakeo, S. Role of complex II in anaerobic respirationof the parasite mitochondria from Ascaris suum andPlasmodium falciparum . Biochim. Biophys. Acta,
1553, 123-139, 2002.
44. Nihei, C., Fukai, Y. and Kita, K. Trypanosome
Alternative Oxidase as a Target of Chemotherapy.
Biochim. Biophys. Acta, 1587, 234-239, 2002.
45. Watanabe, Y., Yokobori, S., Inaba, T., Yamagishi, A.,
Oshima, T., Kawarabayasi, Y., Kikuchi, H. and Kita,
K. Introns in protein-coding genes in Archaea. FEBS
Lett. 510, 27-30, 2002.
46. Miyadera, H., Kano, K., Miyoshi, H., Ishii, N.,
Hekimi, S. and Kita, K. Quinones in long-lived clk-1
mutants of Caenorhabditis elegans. FEBS Lett. 512,33-37, 2002.
47. Kita, K., and Takamiya, S. Electron transfer complexes in Ascaris mitochondria. Advance in Parasitology.
51, 95-131, 2002.
48. Fukai, Y., Nihei, C., Yabu, Y., Suzuki, T., Ohta, N.,
Minagawa, N., Nagai, K. and Kita, K. Strain-specificdifference in amino acid sequences of Trypanosome

Alternative Oxidase. Parasitol. Int. 51, 195-199,
2002.
49. Takashima, E., Ken, D., Osanai, A., Nara, T., Odaka,
M., Aoki, T., Inaka, K., Haradad, S. and Kita, K.
Characterization of the dihydroorotate dehydrogenase as a soluble fumarate reductase in Trypanosoma
cruzi. Mol. Biochem. Parasitol. 122, 189-200, 2002.
50. Miyadera, H., Shiomi, K., Ui, H., Yamaguchi, Y.,
Masuma, R., Tomoda, H., Miyoshi, H., Osanai, A.,
Kita, K. and Omura, H. Atpenins, potent and specificinhibitors of mitochondrial complex II (succinateubiquinone oxidoreductase). Proc. Natl. Acad. Sci.
USA, in press361

Department ofOrgan Pathophysiology andInternal Medicine
Health Service Center
Outline and Research Objectives
In 1972, Health Service Center was founded under the auspices of the Ministry of
Education and Culture to keep sound physical and mental environment in the University ofTokyo. The director of the Center has been elected from a member of Professors in Facultyof Medicine and dealt with two main services, 1) Primary care for undergraduate, graduate
and postgraduate students as well as Faculty Staff and 2) Health Control strategy. At the
opening of the Center, the main task was restricted to the Student with infectious disease
including tuberculosis and malnutrition and medical check of all students in several aspects.
The Center has three branch offices, in Hongo, Komaba and Kashiwa Campuses from 2001.
Recently, the number of students in the University is enormously increasing and reachedabout 25,000. The University also contains Faculty Staffs about 5,000. Target diseases
include a wide variety of sickness, ranging from acute but minor problems like common
cold, acute gastroenteritis, several accidents during experiment, injuries in sports, insomnia
or neurosis to grave or life-threatening sickness such as malignant neoplasms, AIDS,
Hepatitis C, imported infections from foreign countries, DMI and suicide. Graduate andPostgraduate Students are actually main human resources to produce the Research output inLaboratory. Their burdened life in both hard works to do until midnight and competitive
stress to reach a top-level scientist would cause several serious problems.
Official duties of the Center contain 1) Routine medical care of several problems, 2)
Health Check after entrance exam, 3) Regular Health check of all Faculty Members, 4)Regular Health Check for Students from Abroad, 5) Medical Education Programming forchronic diseases, 6) Regular Health Check for Students and Faculty Members treating hazardous materials or radioisotopes, 7) Issue of Health Check Forms, 8) Health Consultation in
general, 9) Health Check for the candidates of new staffs and so on.

Faculties and Students
Professor and Chair Teruhiko TOYO-OKA, M.D., Ph.D. (1991-)
Associate Professors Yoshio UEHRA, M.D., Ph.D. (1999-)
Tsukasa SASAKI, M.D., Ph.D. (2000-)
Lecturers
Shu UWATOKO, M.D., Ph.D. (1997-)
Jun-ichi SUZUKI, M.D., Ph.D. (1998-)
Takashi ISHIKAWA, M.D., Ph.D. (1998-)
Katsuyuki ANDO, M.D., Ph.D. (1997-)
Miyuki SADAMATSU, M.D., Ph.D. (2000-)
Hisako HIKIJI, M.D., Ph.D. (2001-)
Associates ...............................8

Past Research and Major Accomplishments
From 1976, the author has addressed to the pathophysiology of the most serious and popular problems
in medicine in Japan, heart failure. Precise mechanismof the development is still unknown on advanced

362

heart failure from pressure overload secondary tohypertension, several kinds of valvular stenosis, volume overload due to congenital heart anomaly or
valvular insufficiency or from unidentified etiology
including idiopathic cardiomyopathy. Based on an
exact diagnosis, the treatment should be performed,
considering the clinical setting of the patients.
After finishing Ph.D. course in the laboratory ofProf. Setsuro EBASI, who is one of the most outstanding scientists in muscle biology, Dr. Tomoh MASAKI
and I have purified calcium- activated neutral protease(CANP or calpain) from cardiac muscle for the firsttime and characterized both the biochemical andpathological features. Studies on calpain continued
when I moved to the Department of Cardiology in
San Diego or Physiologisches Institut in Heidelberg
where calpain was found to be involved in the loss ofmyofibrillar proteins in myocardial infarction orplatelet aggregation.

Using in vivo 1H and 31P-NMR, the author has indicated structure abnormalities of sarcolemma as thecause for dilated cardiomyopathy (DCM) that is still
the most grave and incurable heart failure. The hereditary form of DCM due to gene abnormalities of cardiac actin, dystrophin, several sarcoglycans (SG) anddesmin has been reported in human cases. We havediscovered gene deletion of δ-SG in human and hamster with hereditary DCM and identified its breakpoint (Sakamoto et al., Proc.Natl.Acad.Sci.USA 1997;
Tsubata et al., J.Clin.Invest. 2000). Comparing withnormal control, BIO 14.6 strains that develop DCM
following a hypertrophy phase, some, but not all, SGs
except δ-SG remained. On the other hand, TO-2
strains, which develop DCM from an early stage, did
not express any SG at all (Kawada et. al., FEBS Lett.
1999). These results indicate the scheme that the
integrity of cell membrane is normally preserved
when all SG are present, but as demonstrated with
BIO 14.6 hamsters, develops compensatory hypertrophy when SG is partially present, and finally reaches
advanced heart failure, when all SGs were lost.
Results from northern blot analysis confirmed abundant mRNA expression of α−, β−, and γ-SGs, andtheir promoter structures were preserved.
Accordingly, we concluded that the actual cause of
DCM is the gene mutation of δ-SG and loss of otherSGs were originated from post-translational level.
To modulate translation of specific mRNA to protein, we demonstrated that transgene expression ismost effectively inhibited with CMV promoter-activated antisense DNA plasmid, rather than general antisense DNA (Chen et al., J.Biol.Chem. 2000; Wang etal., Circ.Res., 2001). In addition, with approval of ethical committee, we conducted genetic diagnosis of
mitochondrial cardiomyopathy and identified a novel
DCM-linked D-loop (Shin et al., Am.J.Hum.Genet.,
2000).
Since the discovery of dystrophin, heart failure
due to dystrophin-related proteins (DRP) abnormalities have been regarded as accompanying symptomsof congenital muscular dystrophy. Although there wasa report on proteolysis of dystrophin following viral
myocarditis, we have reported most cases of acquired
DRP disruption observed in chronic heart failure models were caused by the direct degradation of DRP complex including catecholamine toxicity and heart failure of the old myocardial infarction.
To develop gene therapy of advanced heart failure,
we employed several vectors. Using proteoliposome,
we have transfected eNOS gene to cardiac muscles invivo, and succeeded initiating apoptosis (Kawaguchi
et al., Circulation 1997). We have also indicated thenecessity to employ other vectors for the efficient,
long-term, and biologically inert transfection (Kawada
et al., BBRC, 1999). Our attempt to transduce thecausative gene when it is identified has shown that
responsible gene does improve pathophysiology andrescue the hereditary DCM (Kawada et al., PNAS
2002).

Current Research
Although the prognosis of advanced heart failure
typified by DCM is now gradually improving throughvarious pharmaceutical agents, the final choice isheart transplantation. However, there are still many
ethical, social and medical obstacles such as keeping
the number of donor hearts, and continuing immunosuppressant therapy after the transplantation duringwhole life. The development of gene therapy is anticipated as an ideal method to overcome these problems. As a solution for these problems, we proposed
the following 3-year-plan to develop new strategy forthe prevention and treatment of advanced heart failure (2002 to 2004).
• Research and Survey Projects in the Ministry of
Welfare to investigate the patient population of
the mutated gene previously cloned (2002).
• Development of new potential gene-delivery
system. Using safe and long-lasting adenoassociated virus (rAAV) as a vector, we previouslytransferred responsible gene with normal
sequence, via thoracotomy to above animal model,
which improved the prognosis. To examine if thismethod is clinically applicable to larger subhumananimals, as a step towards clinical use of the basic
research, we will conduct preclinical studies onmonkeys (2002-2004). Specifically, after initiatinglarge-scale production of rAAV (2002), and simultaneously develop administration methods withhigh transfection efficiency. (2003)• Search for more realistic gene-delivery route.
Clinical situation of the patients with advanced
heart failure, however, requires less aggressiveroute not under open chest surgery. Anotheradministration route via transcoronary route
might be preferable to attain the efficient and safedelivery (2004).
• Strict evaluation of safety in regards to generalpathology, immunology, and germ-line cell
infection. Trail of the rAAV administration toskeletal muscle cells is similar but more easilytransfectable than myocardial cells (2002-2003).
• Creation of advanced heart failure presenting
human-type DCM in larger animals. Small animal model does not always represent human diseases. Rhesus monkeys will be employed, because
the Ministry of Welfare in Japan has accepted thisspecies in a preclinical study (2003). Animal models of human-like DCM will be highly innovative
for the development of various disease treatments.

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• Cell transplantation combined with the genetherapy. Patients seek treatment only when
symptoms become severe. Gene therapy waseffective to prevent the disease progression but
could not regenerate the normal cells.
Transduction of normal sequence transgene to cultured muscle cells and create normalized cells in
vitro might be promising for the patients after the
onset of the symptoms. Fortunately, skeletal
myoblasts can proliferate and be transplantable tocardiac tissue and show new development of cardiac muscle cells. Transplantation of geneticallynormalized myoblasts will be promising for the
next generation gene therapy.

Future Prospects
Present project consists of two sections, as follows.
• Double vector strategy for the development ofknockout heart and novel therapy with monkeys. Previous report that knockout of δ-SG genein mice caused not only DCM but also coronary
spasm (Coral-Vasquez et al., Cell 1999). The contribution of myocardial ischemia is of great significance for the pathogenesis of DCM, although they
did not show clear evidence to support thescheme. Because of the limitation of handling insmall heart, no group has succeeded in the verification of the microvessel spasm. To clarify thecontribution of coronary spasm, we are trying toprepare not transgenic mice but knockout heart inlarge animals like Rhesus, to make it possible tovisualize coronary spasm by coronary angiogrrraphy and anaerobic metabolism as follows. AAV
consists of 5 subtypes, Serotype I to V.
Preliminary experiment showed that type V was
the most efficiently transfected both skeletal andcardiac muscle cells in vivo, followed by type III
and II. We have been verifying in vivo since lastyear and succeed in modifying the rAAVs for gene
vector. The rAAV type II as gene therapy vector,
known to be safe and to possess a long expression
period. In addition to type II, we have already prepared type V and are now making sure the safety.
(2002).
Since the serotype of these rAAV vectors completely differ, mutual interference does not take place.
This difference of serotypes has crucial meaning toour study. By applying different rAAV vectors at theinitial and secondary administration, different antigenicity does not prevent double transfection. In ourpresent study, we will initially administer rAAV type
V that is the most efficient with δ-SG antisense,
extensively to monkey cardiac muscle in vivo, andcreate a cardiospecific δ-SG deficient heart. Following

364

validation of transfection via myocardial biopsy using
specific antibody to δ-SG, the pathogenesis will be
precisely examined whether they develop human-likeDCM (2003).
Subsequently, expression of δ-SG containing normal or abnormal sequence can be examined, usingrAAV type II. With the former gene, the rescue will be
expected, as demonstrated previously with hamsters.
With the latter animals, we will confirm the presenceof transgene, and examine the persistence of clinicalsymptoms of dilated cardiomyopathy by physiologicalexaminations by hemodynamics, echocardiography
and assay of BNP (2004).
• Combination therapy of gene and cell therapy at
the advanced heart failure.
Cell therapy will be promising for the treatment ofadvanced heart failure after progression to the degenerative process of cardiomyocytes. Simple cell transplantation tried in humans is not applicable to thecase with gene-deletion or mutation, because intrinsic
gene mutation universally occurs in all tissues including cardiac muscle and skeletal muscle. After skeletal
myoblasts in culture were proliferated and transfectedby normal gene, the treated and normalized muscle
cells can be transplanted to DCM heart. For ethicalconsiderations, genetic diagnosis a priori is essential
for identifying responsible gene or at least, actually
aggravating factor and for developing gene therapy. In
accordance with recommendations by Helsinki
Declaration, Science Council, Ministry of Education,
Ministry of Health and Welfare and related 8
Academic Societies, we requested for a strict inquisition by ethical committee. As stated above, we havealready demonstrated serious consideration to privacy
protection in our previous study on nation-wide
genetic diagnosis survey for the prevention of sudden
death and obtained very meaningful results (Shin etal., Am.J.Hum.Genet. 2000).

Research Grants from 1998 to 2002.
1998-2000 The Ministry of Education, Culture,
Science and Sports; Section A (1), “Novel gene therapyof dilated cardiomyopathy secondary to disruption ofdystrophin-related proteins”.
1998-2000 The Ministry of Education, Culture,
Science and Sports; Section A (2), “Prevention trial of
sudden death with using gene-based presymptomatic
diagnosis”.
2001 Uehara Memorial Foundation, “Developmentof gene-based therapy for dilated cardiomyopathy”.
2002-4 The Ministry of Education, Science, Culture
and Sports; Section A (1), “Gene or cell-engineered
therapy of advanced heart failure”.
2002-4 The Ministry of Health, Labor and Welfare;Translational Research Project from Basic Medicine toClinical Therapy, “Clinical application of gene therapy
with cell engineering for the treatment of advanced
heart failure”.

Select Publications
1. Ebashi S, Nonomura Y, Toyo-oka T. and Katayama E.
Regulation of muscle contraction by the calcium-troponin-tropomyosin system. Cambridge University
Press, Symposia of the Society for Experimental
Biology 30, 349-360, 1976.
2. Toyo-oka T, Shimizu T. and Masaki T. Inhibition ofproteolytic activity of calcium activated neutral proteasebyleupeptin
andantipain.
Biochem.Biophys.Res.Commun. 82, 484-491, 1978.
3. Toyo-oka T. Increased activity of intramuscular proteases in the hyperthyroid state. F.E.B.S.Lett. 117,
122-124, 1980.
4. Toyo-oka T. Calcium ion-insensitive contraction ofglycerinated porcine cardiac muscle fibers by Mg-inosine triphosphate. Circ.Res. 49, 1350-1355, 1981.
5. Toyo-oka T. and Ross JJ. Ca2+ sensitivity change andtroponin loss in cardiac natural actomyosin after
coronary occlusion. Am.J.Physiol. 240, H704-H708,
1981.
6. Toyo-oka T. Phosphorylation with cyclic adenosine 3’
5’ monophosphate-dependent protein kinase renders
bovine cardiac troponin sensitive to the degradationbycalcium-activatedneutralprotease.
Biochem.Biophys.Res.Commun. 107, 44-50, 1982.
7. Toyo-oka T, Nagayama K, Umeda M, Eguchi K. and
Hosoda S. Rhythmic change of myocardial phosphatemetabolite content in cardiac cycle observed bydepth-selected and EKG-gated in vivo 31P-NMR spectroscopy
in
a
whole
animal.
Biochem.Biophys.Res.Commun. 135, 808-815, 1986.
8. Ikeda U, Toyo-oka T. and Hosoda S. Identification of
two new calcium dependent hydrolases in thehuman heart. Biochem.Biophys.Res.Commun. 136,
773-777, 1986.
9. Toyo-oka T, Nagayama K, Umeda M, Ogawa S, Eguchi
K, Hosoda S. and Sugimoto T. Combined study of 1Hmagnetic resonance imaging and depth-selected,
EKG-gated 31P-magnetic resonance spectroscopy ofthe heart in vivo. Biochem.Biophys.Res.Commun.
144, 841-848, 1987.
10. Toyosaki N, Toyo-oka T, Natsume T, Katsuki T,
Tateishi T, Yaginuma T. and Hosoda S. Combination
therapy with diltiazem and nifedipine in patientswith effort angina pectoris. Circulation 77, 13701375, 1988.
11. Toyo-oka T. A critical review of NMR imaging andspectroscopy for the evaluation of cardiac hypertrophy in humans and experimental animals.
J.Mol.Cell.Cardiol. 5, 141-147, 1989.
12. Toyo-oka T, Arisaka H, Sanma H, Shin WS, Dan Y.
and Sugimoto T. Synergistic deleterious effect of
micromolar Ca ions and free radicals on respiratory
function of heart mitochondria at cytochrome C andits salvage trial. Biochem.Biophys.Res.Commun. 163,1397-1403, 1989.
13. Toyo-oka T, Arisaka H, Takayasu T, Tezuka T. and
Hosoda S. In vivo 31P-NMR spectroscopy of heart inhypoxia and identification of biochemical factors
responsible for contractility loss. Prog.Clin.Biol.Res.
315, 567-579, 1989.
14. Toyo-oka T, Shin WS, Okai Y, Dan Y, Morita M,
Iizuka M. and Sugimoto T. Collagen-stimulated
human platelet aggregation is mediated by endogenous calcium-activated neutral protease. Circ.Res. 64,
407-410, 1989.
15. Takayasu T, Toyo-oka T. and Hosoda S. Waste of
ATP for tension development in myocardial acidosis:
Chemomechanical uncoupling at myofibrillar level.
J.Mol.Cell.Cardiol. 22, 127-130, 1990.
16. Naitoh T, Toyo-oka T. and Sugimoto T. An endogenous Ca2+ channel agonist, endothelin-1, does notdirectly activate partially purified dihydropyridinesensitive Ca2+channel from cardiac muscle in a reconstituted system. Biochem.Biophys.Res.Commun. 171,1205-1210, 1990.
17. Okai-Matsuo Y, Takano-Ohmuro H, Toyo-oka T. and
Sugimoto T. A novel myosin heavy chain isoform invascular smooth muscle. Biochem. Biophys. Res.
Commun. 176, 1365-1370, 1991.
18. Shin WS, Toyo-oka T, Masuo M, Okai Y, Fujita H.
and Sugimoto T. Subpopulations of rat vascularsmooth muscle cells as discriminated by calcium
release mechanisms from internal stores. Circ.Res.
69, 551-556, 1991.
19. Masuo M, Toyo-oka T, Shin WS. and Sugimoto T.
Growth-dependent alterations of intracellular Ca2+handling mechanisms of vascular smooth muscle
cells. Circ.Res. 69, 1327-1339, 1991.
20. Suzuki T, Toyo-oka T, Shin WS, and Sugimoto T.
Cell growth-dependent expression of endothelin-1
provocable Ca2+ channels in cloned vascular smoothmuscle cells. J.Cardiovasc. Pharmacol. 17 (Suppl. 7),
S187-S189, 1991.
21. Toyo-oka T, Aizawa T, Suzuki N, Hirata Y, Miyauchi
T, Shin WS, Yanagisawa M, Masaki T. and Sugimoto
T. Increased plasma level of endothelin-1 and coronary spasm induction in patients with vasospastic
angina pectoris. Circulation 83, 476-483, 1991.
22. Toyo-oka T. and Sugimoto T. Endothelin: Key tocoronary vasospasm? Circulation 84, 1451-1452,
1991.
23. Shin WS, Sasaki T, Kato M, Hara K, Seko A, Yang
WD, Shimamoto N, Sugimoto T. and Toyo-oka T.
Autocrine and paracrine effects of endotheliumderived relaxing factor on intracellular Ca 2+ ofendothelial cells and vascular smooth muscle cells:
Identification by two-dimensional image analysis in
coculture. J.Biol.Chem. 267, 20377-20382, 1992.
24. Toyo-oka T, Nagayama K, Suzuki J. and Sugimoto T.
Noninvasive assessment of cardiomyopathy development with simultaneous measurement of topical 1Hand 31P-magnetic resonance spectroscopy. Circulation
86, 295-301, 1992.

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25. Toyo-oka T, Wang YP, Inukai M. and Sakamoto A.
An endothelin family. Recent Progress in Basic andClinical Sciences. Coronary Artery Dis. 2 (Review inDepth), 65-71, 1993.
26. Sakamoto A, Yanagisawa M, Sawamura T, Enoki T,
Ohtani T, Sakurai T, Nakao K, Toyo-oka T. andMasaki T. Distinct subdomains of human endothelin
receptors determine their selectivity to endothelinAselective antagonist and endothelinB-selective agonists. J.Biol.Chem. 268, 8547-8553, 1993.
27. Yang W, Ando J, Korenaga R, Toyo-oka T. andKamiya A. Exogenous nitric oxide inhibits proli-feration of cultured vascular endothelial cells.
Biochem.Biophys.Res.Commun. 203, 1160-1167,
1994.
28. Uehara Y, Hirawa N, Kawabata Y, Suzuki T, Ohshima
N, Oka K, Ikeda T, Goto A, Toyo-oka T, Kizuki K.
Long-term infusion of kallikrein attenuates renal
injury in Dahl salt-sensitive rats. Hypertension 24,
770-778, 1994.
29. Korenaga R, Ando J, Tsuboi H, Yang W, Sakuma I,
Toyo-oka T. and Kamiya A. Laminar flow stimulatesATP- and shear stress-dependent nitric oxide production in cultured bovine endothelial cells.
Biochem.Biophys.Res.Commun. 198, 213-219, 1994.
30. Sakamoto A, Yanagisawa M, Tsujimoto G, Nakao K,
Toyo-oka T. and Masaki T. Pseudo-noncompetitive
antagonism by BQ123 of intracellular calcium transients mediated by human ETA endothelin receptor.
Biochem.Biophys.Res.Commun.200, 679-686, 1994.
31. Kato M, Takazawa K, Kimura A, Ruegg JC, Amano K,
Wang Y, Sakaki Y. and Toyo-oka T. Altered actin
binding with myosin mutation in hypertrophic cardiomyopathy and sudden death. Lancet 345, 1247,
1995.
32. Shin WS, Kawaguchi H, Sasaki T, Wang YP, Yang
WD, Inukai M. and Toyo-oka T.. The role of nitricoxide in cardiovascular system. Ann.N.Y.Acad.Sci.
786, 233-244, 1996.
33. Wang YP, Shin WS, Kawaguchi H, Inukai M, Kato M,
Sakamoto A, Uehara Y, Miyamoto M, Shimamoto N,
Korenaga R, Ando J. and Toyo-oka T. Contribution ofsustained Ca2+ elevation for nitric oxide production inendothelial cells and subsequent modulation of Ca2+transient in vascular smooth muscle cells in coculture. J.Biol.Chem. 271, 5647-5655, 1996.
34. Toyo-oka T. and Naylor WG. Third generation calcium entry blockers. Blood Pressure 5, 206-208, 1996.
35. Kawaguchi H, Shin WS, Wang Y, Inukai M, Kato M,
Matsuo-Okai Y, Sakamoto A, Kaneda Y, and Toyooka T. Apoptosis-like myocardial cell death withoverexpression of human ecNOS gene-Identification
by Sendai virus-coated liposomes and in vivo genetransfection-. Circulation 95, 2441-2447, 1997.
36. Sakamoto A, Ono K, Abe M, Jasmin G, Eki T,
Murakami Y, Masaki T, Toyo-oka T, and Hanaoka, F.
Both hypertrophic and dilated cardiomyopathy arecaused by mutation of the same gene, δ-sarcoglycan,
in hamster. Proc.Natl.Acad.Sci.USA. 94, 1387313878, 1997.

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37. Iwasawa K, Nakajima T, Hazama H, Goto A, Shin
WS, Toyo-oka T and Omata M. Effects of extracellular pH on receptor-mediated Ca2+ influx in A7r5 rat
smooth muscle cells. J.Physiol. 503, 237-251, 1997.
38. Nagoshi H, Uehara Y, Kanai F, Maeda S, Ogura T,
Goto A, Toyo-oka T, Esumi H, Shimizu T and Omata
M. Prostaglandin D2 inhibits inducible nitric oxide
synthase expression in rat vascular smooth muscle.
Circ.Res. 82, 204-209, 1998.
39. Shimamoto R, Suzuki J, Nishikawa J, Fujimori Y,
Nakamura F, Shin WS, Tomaru T and Toyo-oka T.
Measuring the diameter of coronary arteries on MR
angiograms using spatial profile curves. Am.J.Roent.
170, 889-893, 1998.
40. Suzuki J-i, Shimamoto R, Nishikawa J, Fujimori Y,
Nakamura F, Shin WS, Tomaru T and Toyo-oka T.
Vector analysis of the hemodynamics of atherogenesis in the human thoracic aorta with use of MR
velocity mapping. Am.J.Roent. 171, 1285-1290, 1998.
41. Suzuki J-i, Shimamoto R, Nishikawa J, Yamazaki T,
Tsuji T, Nakamura F, Shin WS, Nakajima F, Toyooka T, Ohotomo K. Morphological onset and early
diagnosis in apical hypertrophic cardio- myopathy: along-term analysis with nuclear magnetic resonanceimaging. J.Am.Coll.Cardiol. 33, 146-151, 1999.
42. Kawada T, Shin WS, Nakatsuru Y, Koizumi T,
Sakamoto A, Nakajima T, Nakazawa M, Sato H
Takato T, Ishikawa T and Toyo-oka T. Precise identification of gene-products in hearts after in vivo genetransfection, using Sendai virus-coated proteoliposomes. Biochem.Biophys.Res.Commun. 259, 408413, 1999.
43. Hirawa N, Uehara Y, Kawabata Y, Numabe A, Gomi
T, Ikeda I, Suzuki T, Goto A, Toyo-oka T and Omata
M. Long-term inhibition of renin-angiotensin system
sustains memory function in aged Dahl rats.
Hypertension 34, 496-502, 1999.
44. Kawada T, Nakatsuru Y, Sakamoto A, Koizumi T,
Shin WS, Okai-Matsuo Y, Suzuki J, Uehara Y,
Nakazawa M, Satoh H, Ishikawa T and Toyo-oka T.
Strain- and age-dependent loss of sarcoglycan complex in cardiomyopathic hamster hearts and its reexpression by d-sarcoglycan gene transfer in vivo.
F.E.B.S.Lett. 458, 405-408, 1999.
45. Shin WS, Tanaka M, Suzuki J, and Toyo-ok T. A
novel homoplamic mutation in mtDNA with a single
evolutionary origin as a risk factor for cardiomyopathy. Am.J.Hum.Genet. 67, 1617-1620, 2000.
46. Chen J, Wang YP, Nakajima T, Iwasawa K, Hikiji H,
Sunamoto M, Yoshida Y, Sakaki Y, and Toyo-oka T.
Autocrine action and its underlying mechanism of
nitric oxide on intracellular Ca2+ homeostasis in vascular endothelial cells. J.Biol.Chem. 275,2873928749, 2000.
47. Wang YP, Chen J, Wang Y, Taylor CW, Hirata Y,
Hagiwara H, Mikoshiba K, Toyo-oka T, Omata M,
and Sakaki Y. Crucial role of type 1, but not type 3,
IP3 receptors in IP3-induced Ca2+ release, capacitative
Ca2+ entry and proliferation of A7r5 vascular smoothmuscle cells. Circ.Res. 88, 202-299, 2001.

48. Kawada T, Sakamoto A, Nakazawa M, Urabe M,
Masuda F, Hemmi C, Wang Y, Shin WS, Nakatsuru
Y, Sato H, Ozawa K and Toyo-oka T. Morphologicaland physiological restorations of hereditary form of
dilated cardiomyopathy by somatic gene therapy.
Biochem.Biophys.Res.Commun. 284, 431-435, 2001.
49. Shin WS. Hemmi C and Toyo-oka T. Coculture andcross talk between endothelial cells and vascularsmooth muscle cells mediated by intracellular Ca2+.
Methods in Molecular Biology, Ed. C. Wise, Medical
Research Council Press (London), 347-357, 2001.
50. Kawada T, Nakazawa M, Nakauchi S, Yamazaki K,
Shimamoto R, Urabe M, Nakata J, Hemmi C, Masui
F, Nakajima T, Suzuki J, Monahan J, Sato H, Takeo
S, Ozawa K and Toyo-oka T. Rescue of hereditary
form of dilated cardiomyopathy by rAAV-mediatedsomatic gene therapy.Proc.Natl.Acad.Sci.USA. 99,
901-906, 2002.

367

Radiation
Research Institute
Outline and Research Objectives
Radiation Research Institute (formerly called as Radioisotope Research Institute) wasestablished in 1992 for the purpose of radiation safety control of five radioisotope researchlaboratories belonging to Faculty of Medicine. They are located in basic science buildings,
Animal Center, University Hospital, and University Branch Hospital. (The laboratory in
University Branch Hospital was closed in March, 2002.) The research is also expected to theInstitute on the subjects related to radiations and radioisotopes such as radiation biology,
medical application of radiations/radioisotopes, and health physics.

Faculties and Students
Professor and Chair

Kunio Shinohara, Dr. of Medical
Science (1996-)
Lecturer
Takahiko Suzuki, Dr. of
Pharmaceutical Science
Research Student ....................1

Past Research and Major Accomplishments
I (K. Shinohara) learned applied physics at undergraduate course. After I got masters degree in biophysics, I took a doctoral course in medical science. In
the field of radiation biology, I started my research
work on the cytotoxic effects of γ-rays on cultured
mammalian cells in a colony state that is a model system for tumor cord [1]. Then the subject moved to
DNA damage and its repair induced by radiation andradiomimetic chemicals such as methylazoximethanol
acetate, an active principle of cycasin which is naturally occurring carcinogen in cycad, and benzo(a)pyrene.
When new radiation source, synchrotron radiation,
became available, our research subjects shifted tofocus on biological/medical application of monochromatic X-rays such as Auger enhancement of radiationinduced cell death and development and applicationof soft X-ray microscopy. In the mean time we foundthat to image hydrated specimens by soft X-ray
microscopy at high resolution, we need intense X-ray
source to image at a shorter time than the movement
by Brownian motion. For this purpose, we appliedlaser-produced plasma X-rays, by which the exposure
time was as short as 300 psec.
1. DNA damage and its repair induced by radiation and radiomimetic chemicals
With the aid of caffeine, we found that methylazoxymethanol acetate induces damage, which isrepaired by a mechanism analogous to post-replication repair of UV light-induced damage [3]. In this368

study, we used synchronized cells obtained by theautomatic synchronizer which we developed [2].
In the study of another carcinogen,benzo(a)pyrene, we detected the formation and excision repair of benzo(a)pyrine-DNA adducts in mammalian cells [4, 6]. This adducts were also detected inperipheral human lung tissue when the tissue wascultured in the presence of benzo(a)pyrene [5].
It is also observed that depression level of DNA
synthesis is well correlated to the sensitivity of cell
death to UV-irradiation in normal human cells and incells derived from xeroderma pigmentosum patient
deficient in DNA repair [8].
2. Radiation- and heat-induced cell death(1) Heat-induced cell death
Hyperthermia is developed as one of the method
for cancer treatment. The sensitivity of cell death
induced by the heat-treatment was studied withrespect to energy metabolism. It has been demonstrated that when the synthesis of ATP is inhibited,
sensitivity of the cell to heat increases [16]. Our
results show that ATP level decreases after heat-treatment and in a treatment-time dependent manner [15].
The sensitivity of cells to heat is well correlated to the
ATP level of the cells at normal growth condition
which corresponds to the pool size of ATP (or potential to synthesize ATP) [26,49]. The results indicatethat energy metabolism is a good marker to evaluate
heat sensitivity. We also observed that Ga citrate, atumor-seeking compound, is a good sensitizer to heat
treatment [17,47].
(2) Studies on radiation-induced cell death
X-rays inhibit DNA synthesis. Initiation of DNA
synthesis is much more sensitive to X-rays than its
elongation. We studied carefully this inhibition andresumption of replicon initiation after X-irradiation[7].
Synchrotron radiation is an intense radiation,

intense enough to study the effects of monochromatic
X-rays. We first studied an action spectrum of lethal
effects on mammalian cells in the wavelength range
from 160 to 254 nm. Cells were sensitive to 160 nm
and 254 nm and relatively resistant to 190 nm [9]. We
found that target site was not DNA but membrane at
the wavelength of 160 nm [13].
(3) Auger enhancement
When elements absorb X-rays at the inner shellelectrons, they release Auger electrons and are highly
toxic. Their LET corresponds to 5 MeV α particles(100 keV/µm). Therefore, efficient induction of Auger
effects in cells by X-rays will result in the sensitization to the cells. We studied this enhancement bylabeling the cells with 5-bromodeoxyuridine or 5iododeoxyuridine and monochromatic synchrotron
radiations at the wavelengths slightly higher than K
absorption edges of these elements (i.e., Br and I). Our
results are the first data for the Auger enhancementof cytotoxicity on mammalian cells obtained by synchrotron radiation [10,11]. Auger enhancement is also
observed in the induction of chromosome aberration[18]. Theoretical reports pointed the great enhancement and suggested the new possible modality forradiation therapy. However, our experimental results
were not in accord with this theory [11]. We re-evaluated this theoretical consideration and found that thetheory needs correction. Our correction of the theory
fits well to the experimental data [14]. This report[14] is cited in News and Views in Nature by John
Humm (336, 710-711, 1988). The results were against
the expectation for radiation therapy, but this is science. In the same issue of Nature, another method ofAuger enhancement was reported. The method basedon the nuclear resonance absorption of the element.
Although the results presented in the report could
have not been confirmed yet, the idea is attractive.
We estimated the enhancement ratio using the same
equation as above and found that more than 10 times
higher enhancement will be expectable [unpublishedresults]. However, it is hard to obtain the experimental data to evaluate this theoretical estimate at thepresent stage. To keep the potential to study Auger
enhancement, we tried to detect and characterize
Auger enhancement using laboratory X-ray source.
We observed small fraction of Auger enhancementthat was protectable by cysteamine as a proton donor
for radiation damage [41]. We also studied suicide
experiment with a radioisotope of iodine-125 [28].
(4) Molecular mechanisms of radiation-induced
cell death
Radiation-induced cell death has been classified
into two groups: reproductive death and interphase
death. Reproductive death is determined by thecolony forming ability, while interphase death isthought to be functional loss and estimated by loss ofmembrane function (as demonstrated by the dye
exclusion test) or energy metabolism. We first studied
the difference in these two modes of cell death. For
this purpose human leukemic MOLT-4 cells are well
suited. They show interphase death at relatively low
dose irradiation and also are able to be determined forthe colony forming ability. The data [29] show thatboth types of cell death occur in MOLT-4 cells andthat the mechanism of cell death is apoptosis. Next,
we compared MOLT-4 with M10, which is a radiosensitive mutant of L5178Y derived from mouse lymphocytic leukemia and has the same level of radiosensitivity. Unexpectedly, we found that M10 developed
necrosis, while MOLT-4 died by apoptosis at the same
dose level of irradiation [31]. It is confirmed with theheat-treatment that M10 has a system to induce apoptosis (, which is not working at X-irradiation,) probably by the different mechanism from MOLT-4 [31,42].
Based on these results, we are interested in themolecular mechanisms of radiation-induced apoptosis(MOLT-4) and necrosis (M10). Now we are focused on
the apoptosis in MOLT-4 cells. (Because of the lack of
manpower, radiation-induced necrosis in M10 isremained to be studied in future.) Radiation-induced
apoptosis in MOLT-4 cells is p53 dependent [45,48]
and caspase-3 dependent [50]. It should be noted that
inhibition of proteasome also induced apoptosis in
MOLT-4 cells probably with the different mechanism
from X-rays because both apoptosis were additive[39].

3. Development and application of soft X-ray
microscopy
Soft X-rays in a wavelength range of 1-10 nm areprovided by synchrotron radiation and moderately
absorbed by biological materials. Soft X-ray
microscopy has following advantages over optical andelectron microscopy: higher penetration depth thanelectron microscopy and higher resolution than optical microscopy. Therefore, with soft X-ray
microscopy, it is expected that we can observe awhole live cell at higher resolution than optical
microscopy. When we started this project, there wasno soft X-ray microscope. And now, internationally,
there are some microscopes with the potential toimage at a resolution of 20 nm. However, unfortunately, no active microscope at high resolution has beenworking in Japan. This is the present stage. Morethan 15 years ago, we were interested in soft X-ray
microscopy and started to study the importance of
this new method in biology.
When we started, the only method available forexperimental test was a contact microscopy using a369

thin film of polymethylmetacrylate (PMMA) that isdeveloped and characterized by IBM for industrial
purpose. We studied and applied it for imaging
human cells and chromosomes with some improvement of the method for the observation of PMMA bya transmission electron microscope [12]. We have succeeded in the observation of chromosome fibers andnucleosomes [19-21,33]. The results (Fig. 1) were top
data with respect to the resolution to image biologically meaningful specimen and were cited in books from
USA, China and Japan. Then we studied the condition
to image hydrated biological specimens including the
problem of radiation damage and image blurring byBrownian motion [22,24,25]. The conclusion of thisstudy proposed the use of short pulsed X-rays forimaging intact specimens in hydrated condition withsuccessful results for hydrated chromosomes with asingle shot exposure of laser-produced plasma X-rays
at the exposure time of 300 picoseconds [23,30,32].
However, our results were obtained by contact
microscopy and the method is good for molecules but
not well suited for imaging inside a whole cell.
Therefore, we proposed a possible microscope system
to image whole cells [27]. The system is now in thestage of development by other groups.
Another important factor for the application ofsoft X-ray microscopy to biological/medical science isthe contrast of the images for thick specimens. To
overcome this problem, specific staining with threedimensional observation may be the best choice. In
the case of soft X-rays, there is another possibility toidentify elements and molecules: spectroscopic analysis of imaging (spectromicroscopy). We have beenworking on spectromicroscopy in two aspects: instrumentation [43,44,46] and application [34,37]. This lineof work is in progress. For three-dimensional imaging,
we have also studied on holographic microscopy

Fig. 1. X-ray image of a stretched portion of a human chromosome [21]. An enlargement of the area indicated by an arrow in(a) is presented in (b).

370

[35,36,38,40] and found that even with holographic
observation, three dimensional reconstruction may
not be possible with a single exposure.

Current Research
1. Radiation-induced cell death(1) Molecular mechanisms of radiation-induced
apoptosis
Our present study is focussed on the switchingmechanism of p53 to select DNA repair or to decidesuicide. For this study we are currently collecting thedata for p53, and related molecules and phenomenain each single cell to get the idea what is going on in
an individual single cell.
(2) Auger enhancement
As shown above, theoretical consideration showsthat Auger enhancement following nuclear resonanceabsorption is an attractive method. Although we havean idea to evaluate the theoretical results experimentally, we have no clue to apply this method for clinicaltumor therapy. Therefore, the work remains in theoretical level at the present stage.
(3) Cytotoxic effects of ultrahigh dose rate X-rays
Time scale of radiation effects on life begins withphysical interaction (10-18-10-15 sec) followed by chemical and biological processes. At a conventional
method of X-irradiation, these physical and chemical
processes will happen dispersively and induce ionization as one after another. In contrast, particle beamdensely ionizes irradiated materials and gives
increased cytotoxicity to cells. It is of interest to see
what will happen if one can concentrate ionization by
X-rays in a short instance especially in shorter than
one picosecond. Since such a radiation source is available in recent years, we started to study the cytotoxic
effects of ultrahigh dose rate X-rays on mammaliancells at the Institute of Laser Engineering, Osaka
University. Because of limited beam time, we are gradually correcting results and have no publication so far,though five years have already passed.

2. Development and application of soft X-ray
microscopy
Currently our subject is focussed on spectromicroscopy of the cell to understand images for the
improvement of contrast. We have collected images ofa cell at various wavelengths in a wide range (1-10nm). With absorption characteristics of elements andmolecules, we are trying to identify these elements
and molecules using computer-assisted image process-ing. The work is in collaboration with Prof. Ito of
School of Engineering, Tokai University.

Future Prospects
1. Radiation-induced cell death(1) Auger enhancement
It is not clear that Auger enhancement can beapplied for radiation therapy. However, I believe it isworth to continue studying.
(2) Ultrahigh dose rate X-rays in biology and medicine
The new radiation source may be a useful methodfor discovering the new mode of diagnostics and radiation therapy. At present the work is in the beginningstage. We will keep working on it.
(3) Molecular mechanisms of radiation-induced
apoptosis and necrosis
We are planning to understand the full scheme of
the flow of signal transduction process in an individual cell from DNA damage induced by X-rays to thedevelopment of apoptosis. The results will contributeto understand the whole scheme of apoptosis and to
explore the new mode of radiation therapy with theinduction of apoptosis.
2. Soft X-ray microscopy
We believe that soft X-ray microscopy will beestablished and applied to biological/medical research
as a complementary method to optical and electron
microscopy in future. The system will become compact and stay in laboratory in the size comparable to atransmission electron microscope.

Research Grants
1. Grant-in-Aid for Scientific Research (B) (1) [20012003] Studies on biological effects of ultrahigh dose
rate pulsed X-rays.
2. Grant-in-Aid for Scientific Research (A) (1) [19972000] Development and its medical/biological application of a projection X-ray microscope using synchrotron radiation.
3. Grant-in-Aid for Exploratory Research [1997-1998]
Studies on cytotoxic effects of ultrahigh dose rate Xrays.
4. Grant-in-Aid for Scientific Research (A) (1) [19951997] High resolution imaging analysis of elements
and molecules in a cell with an X-ray contact microscope.

Select Publications
1. Shinohara, K. and Okada, S.: Radiosensitivities ofmurine lymphoma L5178Y cells in a multi-cellularcolony system. J. Radiat. Res. 13, 109-116, 1972.
2. Okada, S. and Shinohara, K.: An automatic synchronizer for suspension culture of L5178Y cells. Cell
Tissue Kinet. 7, 195-199, 1974.
3. Shinohara, K. and Matsudaira H.: Evidence for caffeine-sensitive damage in methylazoxymethanol
acetate-treated L5178Y cells. Chem.-Biol. Interactions
12, 53-63, 1976.
4. Shinohara, K. and Cerutti, P.A.: Excision repair ofbenzo(a)pyrene-deoxyguanosine adducts in baby
hamster kidney 21/C13 cells and in secondary mouseembryo fibroblasts C57BL/6J. Proc. Nat’l. Acad. Sci.
USA 74, 979-983, 1977.
5. Shinohara, K. and Cerutti, P.A.: Formation of
benzo(a)pyrene-DNA adducts in peripheral humanlung tissue. Cancer Letters 3, 303-309, 1977.
6. Feldman, G., Remsen, J., Shinohara, K. and Cerutti,
P.A.: Excisability and persistence of benzo(a)pyrene
DNA adducts in epithelioid human lung cells. Nature
274, 796-798, 1978.
7. Shinohara, K. and Watanabe, I.: Inhibition andresumption of replicon initiation in X-irradiatedmouse L5178Y cells. Radiat. Environ. Biophys. 21,
163-173, 1983.
8. Shinohara, K.: Lethality and the depression of DNA
synthesis in UV-irradiated normal human and xeroderma pigmentosum cells. Mutation Res. 122, 385389, 1983.
9. Shinohara, K., Ito, T., Ito, A., Hieda, K., Kobayashi, K.
and Okada, S.: An approach to the use of synchrotron radiation for investigating radiation effects on
cultured mammalian cells in the range from 160 to
254 nm. Radiat. Res. 97, 211-215, 1984.
10. Shinohara, K., Ohara, H., Kobayashi, K., Maezawa,
H., Hieda, K., Okada, S. and Ito, T.: Enhanced killingof HeLa cells pre-labeled with 5-bromodeoxyuridine
by monochromatic synchrotron radiation at 0.9 Å:An evidence for Auger enhancement in mammaliancells. J. Radiat. Res. 26, 334-338, 1985.
11. Shinohara, K., Ohara, H., Hiraoka, T., Nakano, H.
and Ando, M.: Irradiation of cultured mammaliancells labeled with iododeoxyuridine by monochromatic synchrotron radiation at 0.370 and 0.380 Å.
Photon Factory Activity Report 1984/85, National
Laboratory for High Energy Physics, p. VI-153, 1986.
12. Shinohara, K., Aoki, S., Yanagihara, M., Yagishita, A.,
Iguchi, Y. and Tanaka, A.: A new approach to theobservation of the resist in X-ray contact
microscopy. Photochem. Photobiol. 44, 401-403,
1986.
13. Shinohara, K., Ito, A. and Ito, T.: Cell surface damage
in cultured mammalian cells with synchrotron radiation at 160 nm. Photochem. Photobiol. 44, 405-407,
1986.
14. Shinohara, K., Nakano, H., Hiraoka, T. and Ohara,
H.: Re-evaluation of the theoretical study of photon

371

activation therapy. Photon Factory Activity Report
1986, National Laboratory for High Energy Physics,
p. 278, 1987.
15. Shinohara, K., Kugotani, M., Ishiwata, J. andMatsuda, T.: Early change of adenosine triphosphatelevels in L5178Y cells during hyperthermia. Jpn. J.
Cancer Res. (Gann) 78, 333-335, 1987.
16. Shinohara, K. and Matsuda, T.: Hyperthermia andcellular ATP levels. Radiosensitization Newsletter 6,
9-10, 1987.
17. Shinohara, K., Kawakami, N., Kugotani, M. andNakano, H.: Gallium citrate, a new sensitizer of cells
to hyperthermia. Jpn. J. Cancer Res. (Gann) 79, 2123, 1988.
18. Ohara, H., Shinohara, K., Kobayashi, K. and Ito, T.:
An additional enhancement in BrdU-labelled cultured mammalian cells with monoenergetic synchrotron radiation at 0.09 nm: Auger effect in mammalian cells. In “DNA Damage by Auger Emitters”(Ed. by K. F. Baverstock and D. E. Charlton), Taylor& Fancis, pp. 123-133, 1988.
19. Shinohara, K.: Observation of human chromosomeswith soft X-ray contact microscopy. SPIE 1140, 196200, 1989.
20. Shinohara, K.: Application of X-ray microscopy tomammalian cells. In “X-Ray Microscopy in Biology
and Medicine” (Ed. by Kunio Shinohara, Hiroshi
Kihara, Takuma Saito and Keiji Yada), Japan
Scientific Societies Press/Springer-Verlag, pp. 203212, 1990.
21. Shinohara, K., Nakano, H., Kinjo, Y. and Watanabe,M.: Fine structure of unstained human chromosome
fibres dried with no fixative as observed by X-ray
contact microscopy. J. Microscopy 158, 335-342,
1990.
22. Shinohara, K. and Ito, A.: Radiation damage in soft
X-ray microscopy of live mammalian cells. J.
Microscopy 161, 463-472, 1991.
23. Shinohara, K., Ito, A., Kinjo, Y. Watanabe, M.,
Kikuchi, Y. and Tanaka, K.A.: A simple chamber forhydrated specimen in a vacuum and its application
to X-ray contact microscopy with laser produced
plasma X-rays. In “X-RAY MICROSCOPY III” (Ed. by
A. Michette, G. Morrison and C. Buckley), Springer
Series in Optical Sciences Vol. 67, Springer-Verlag,
pp. 347-349, 1992.
24. Shinohara, K. and Ito, A.: Factors affecting the observation of hydrated mammalian cells by soft X-ray
microscopy. In “X-RAY MICROSCOPY III” (Ed. by A.
Michette, G. Morrison and C. Buckley), Springer
Series in Optical Sciences Vol. 67, Springer-Verlag,
pp. 397-399, 1992.
25. Ito, A. and Shinohara, K.: Image blurring by thermal
diffusion in the observation of hydrated biomolecules with soft X-ray microscopy. Cell Structure andFunction 17, 209-212, 1992.
26. Shinohara, K., Kawakami, N., Ito, A., Kugotani, M.,
Nakano, H., Ishiwata, J. and Matsuda, T.: The role of
ATP metabolism in hyperthermic cell killing. In“Cancer Treatment by Hyperthermia, Radiation and372

Drugs” (Ed. by T. Matsuda), Taylor and Francis, pp.
79-91, 1993.
27. Richardson, M., Shinohara, K., Kinjo, Y., Tanaka,
K.A., Ikeda, H. and Kado, M.: Pulsed X-ray
microscopy of biological specimens with laser plasma
source. SPIE 1741, 133-141, 1993.
28. Miyazaki, N. and Shinohara, K.: Cell killing induced
by decay of 125I during the cell cycle: comparison of
125
I-antipyrine with 125I-bovine serum albumin. Radiat.
Res. 133, 182-186, 1993.
29. Shinohara, K. and Nakano, H.: Interphase death andreproductive death in X-irradiated MOLT-4 cells.
Radiat. Res. 135, 197-205, 1993.
30. Shinohara, K., Ito, A. and Kinjo, Y.: X-ray microscopyof biological specimens with laser plasma X-rays.
SPIE 2015, 10-19, 1994.
31. Nakano, H. and Shinohara, K.: X-ray-induced cell
death: apoptosis and necrosis. Radiat. Res. 140, 1-9,
1994.
32. Kinjo, Y., Shinohara, K., Ito, A., Nakano, H.,
Watanabe, M., Horiike, Y., Kikuchi, Y., Richardson,
M.C. and Tanaka, K.A.: Direct imaging in a waterlayer of human chromosome fibres composed of
nucleosomes and their higher-order structures bylaser plasma X-ray contact microscopy. J. Microscopy
176, 63-74, 1994.
33. Shinohara, K., Yada, K. and Kinjo, Y.: X-ray
microscopy of chromosome structures. In “X-ray
Microscopy IV” (Ed. by A. I. Erko and V. V. Aristov),
Bogorodskii Pechatnik Publishing Company, pp. 213216, 1994.
34. Shinohara, K., Ito, A., Matsumura, T., Kinoshita, K.
and Kobayashi, K.: Absorption spectra at the wavelength range of 1.5-10 nm and element-imaging of
cultured mammalian cells with electronic zooming
tube. In “X-ray Microscopy IV” (Ed. by A. I. Erko and
V. V. Aristov), Bogorodskii Pechatnik Publishing
Company, pp. 264-267, 1994.
35. Daido, H., Schulz, M.S., Murai, K., Kodama, R., Yuan,
G., Goto, J., Tanaka, K.A., Kato, Y., Nakai, S.,
Shinohara, K., Honda, T., Kodama, I., Iwasaki, H.,
Yshinobu, T., Tsukamoto, M., Niibe, M., Fukuda, Y.,
Neely, D., MacPhee, A. and Slark, G.: Subnano-second in-line soft X-ray holography using germanium
laser in the 20 nm wavelength region. J. X-ray Sci.
Technol. 5, 105-120, 1995.
36. Shinohara, K., Ito, A., Nakano, H., Kodama, I.,
Honda, T., Matsumura, T. and Kinoshita, K.: X-ray
holographic microscopy of biological specimens with
an electronic zooming tube. J. Synchrotron Radiat. 3,
35-40, 1996.
37. Ito, A., Shinohara, K., Nakano, H., Matsumura, T.
and Kinoshita, K.: Measurement of soft X-ray absorption spectra and elemental analysis in local regionsof mammalian cells using an electronic zooming
tube. J. Microscopy 181, 54-60, 1996.
38. Kodama, I., Yamaguchi, M., Ohyama, N., Honda, T.,
Shinohara, K., Ito, A., T., Matsumura, T., Kinoshita,
K. and Yada, K.: Image reconstruction from an in-line

X-ray hologram with intensity distribution constraint. Optics Commun. 125, 36-42, 1996.
39. Shinohara, K., Tomioka, M., Nakano, H., Toné, S.,
Ito, H. and Kawashima, S.: Apoptosis induction
resulting from proteasome inhibition. Biochem. J.
317, 385-388, 1996.
40. Shinohara, K.: Possible applications of X-ray lasers inbiology: X-ray microscopy and X-ray lasers. In “X-ray
Lasers 1996” (Ed. by S. Svanberg and C.-G.
Wahlstrom), Institute of Physics Conference Series
Number 151, Institute of Physics, pp. 533-538, 1996.
41. Shinohara, K., Nakano, H. and Ohara, H.: Detectionof Auger enhancement induced in HeLa cells labeledwith iododeoxyuridine and irradiated with 150 kV
X-rays: Effects of cysteamine and dimethylsulfoxide.
Acta Oncologica 35, 869-875, 1996.
42. Nakano, H., Kurihara, K., Okamoto, M., Toné, S. andShinohara, K.: Heat-induced apoptosis and p53 in
cultured mammalian cells. Int. J. Radiat. Biol. 71,
519-529, 1997.
43. Ito, A., Shinohara, K., Mizukami, Y., Nakano, H.,
Yada, K., Uehara, T. and Honda, T.: X-ray contact
microscopy system for spectromicroscopy of biological specimens. J. Synchrotron Radiat. 5, 1099-1101,
1998.
44. Shinohara, K., Ito, A., Nakano, H., Honda, T. and
Yada, K.: X-ray microscopy system with an electronic
zooming tube. In “X-Ray Microscopy andSpectromicroscopy” (Ed. by J. Thieme, G. Schmahl, E.
Umbach and D. Rudolph), Springer-Verlag, pp. I-169I-173, 1998.
45. Nakano, H. and Shinohara, K.: Correlation between
unirradiated cell TP53 protein levels and radiosensitivity in MOLT-4 cells. Radiat. Res. 151, 686-693,
1999.
46. Shinohara, K., Ito, A., Honda, T., Yoshimura, H. and
Yada, K.: Development of an X-ray projection microscope using synchrotron radiation. In “X-Ray
Microscopy: Proceedings of the Sixth International
Conference” (Ed. by W. Meyer-Ilse, T. Warwick, and
D. Attwood), AIP Conference Proceedings 507,
American Institute of Physics, pp. 346-349, 2000.
47. Miyazaki, N., Nakano, H., Kawakami, N., Kugotani,M., Nishihara, K., Aoki, Y. and Shinohara, K.:
Sensitizing effects of gallium citrate on hyperthermic
cell killing in vitro. Int. J. Hyperthermia 16, 457-470,
2000.
48. Nakano, H., Kohara, M. and Shinohara, K.:
Evaluation of the relative contribution of a p53-mediated pathway in X-ray-induced apoptosis in MOLT-4
cells by transfection with mutant p53 gene at different expression levels. Cell Tissue Res. 306, 101-106,
2001.
49. Miyazaki, N., Kurihara, K., Nakano, H. andShinohara, K.: Role of ATP in the sensitivity to heatand the induction of apoptosis in mammalian cells.
Int. J. Hyperthermia 18, 316-331, 2002.
50. Nakano, H. and Shinohara, K.: Time sequence analysis of caspase-3-independent programmed cell death

and apoptosis in X-irradiated human leukemic
MOLT-4 cells. Cell Tissue Res., 310, 305-311, 2002.

373