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Program / Abstract Book

Asian Society of Clinical Pathology and
Laboratory Medicine
12 th Meeting of the Asian Society of Clinical Pathology and Laboratory Medicine
(ASCPaLM KYOTO 2012)
The Future of Laboratory Medicine in
Clinical Practice
29 November – 1 December 2012
Kyoto International Conference Center
Room D / Event Hall
www3.kmu.ac.jp/ascpalm2012/information.html

Welcome
Dear Friends and Colleagues,
On behalf of the Organizing Committee for the 12th Meeting of the Asian Society of Clinical Pathology and
Laboratory Medicine (ASCPaLM), we take great pleasure in welcoming you to Kyoto.
This is a truly international meeting with participants from all over the world, coming together to hear of the latest
developments in the basic, clinical and population research into clinical pathology and laboratory medicine and
associated conditions. We are sure that you’ll find the excellent program contemporary, stimulating and relevant. The
state-of-the-art reviews presented by the industry-sponsored seminars will provide opportunities to bring yourself right
up-to-date in the diversity of themes outside your expertise. The program incorporates a variety of presentation formats
including Plenary Sessions, Sponsored Symposia, Luncheon Seminar and Poster Sessions that have been structured to
facilitate interaction and discussion.
Networking with colleagues is not just limited to the scientific program. The social program is great time to catchup
with friends and collaborators in a more relaxed atmosphere, while experiencing the best that spectacular Kyoto has to
offer.
We look forward to seeing you during the coming month and would like to take this opportunity to thank you for
attending ASCPaLM KYOTO 2012.
Hakuo Takahashi
Chair, Organising Committee
ASCPaLM KYOTO 2012
President, ASCPaLM
Organizing committee for the ASCPaLM KYOTO 2012
Department of Clinical Sciences and Laboratory Medicine
Kansai Medical University
2-3-1 Shinmachi, Hirakata City, Osaka 573-1191, Japan
TEL/FAX: +81-72-804-2773

Table of Contents
Committees 1
Sponsors and Supporters 3
General Information 3-8
Scientific Program Information 9-11
Program at a glance 12
Keynote lecture 13
Featured Oral Presentation 14
abstracts 15-20
Advanced technical seminars 21-34
Scientific and Poster Sessions (chairperson, title & Speakers) 35-52
Abstracts-Clinical Chemistry 54-74
Abstracts-Endocrinology 75-79
Abstracts-Immunology and Blood Bank 80-87
Abstracts-Hematology 88-104
Abstracts-Clinical Microbiology 105-118
Abstracts-Physiological Function test 119-120
Abstracts-Pathology 121-127
Abstracts-Molecular biology 128-139
Abstracts-Others 140-152
Notes 154-

ASCPaLM 2012 Executive Committee
President Hakuo Takahashi Department of Clinical Sciences and Laboratory
Medicine, Kansai Medical University, Osaka, Japan
Vice president Oh Hun Kwon Department of Laboratory Medicine, Yonsei University
College of Medicine, Seoul, Korea
Secretary General Tjin Shing Jap Section of Biochemistry, Department of Pathology and
Laboratory Medicine, Taipei Veterans General Hospital,
Taiwan
National Representatives (President of the Society)
Mitsuru Murata The Japanese Society of Laboratory Medicine (JSLM)
Won-ki Min The Korean Society for Laboratory Medicine
Lia G. Partakusuma Indonesian Association of Clinical Pathologists
Tjin Shing Jap Taiwan Society of Clinical Pathologists
N. Munkhtuvshin Mongolian Society of Clinical Pathologists
Organisers
Chairperson Hakuo Takahashi Kansai Medical University
Advisory Tadashi Kawai Former President, the JSLM
Board Ikunosuke
Sakurabayashi
Former President, the JSLM
Kiyoaki Watanabe Former President, the JSLM
Yukihisa Miyazawa Former President, the JSLM, Teikyo University
Mitsuru Murata President, The JSLM, Keio University, School of
Medicine
Tomohiro Samori President, the Japanese Association of Clinical Laboratory
Physicians
Satoshi Ichiyama Chairman, the 59 th JSLM annual meeting, Kyoto
University
Yutaka Yatomi Tokyo University, Graduate School of Medicine
Hayato Miyachi Tokai University, School of Medicine
Akiko Yoneyama Toranomon Hospital
Masami Murakami Gunma University, Graduate School of Medicine
Masato Maekawa Hamamatsu University, School of Medicine
Junichi Chihara Akita University, Graduate School of Medicine
- 1 -

Program
Committee
Yukio Ozaki University of Yamanashi, Faculty of Medicine
Akira Suwabe Iwate Medical University, School of Medicine
Isao Kitajima University of Toyama, Graduate School of Medicine and
Pharmacological Sciences
Yukio Ando Kumamoto University, Graduate School of Medical
Sciences
Tsutomu Nobori Mie university Graduate School of Medicine
Hidetoshi Okabe Shiga University of Medical Science
Hiroo Tominaga Kasai City Hospital
Hitosi Asayama Osaka Association of Medical Technologist
Yasuyuki Okamoto Nara Medical University
Shunichi Kumagai Kobe University, Graduate School of Medicine
Masahiro Koshiba Hyogo College of Medicine, School of Medicine
Tokio Sanke Wakayama University of Medical Science
Takayuki Takubo Osaka Medical College
Yoh Hidaka Osaka University Graduate School of Medicine
Naohisa Fujita Kyoto Prefectural University of Medicine
Toshinori Kamisako Kinki University, School of Medicine
Kuniaki Saito Kyoto University Graduate School of Medicine
Shuji Matsuo Tenriyorozu Hospital
Shunji Takakura Kyoto University Graduate School of Medicine
Takahiro Doi Kyoto University Graduate School of Medicine
Yutaka Furukawa Kobe City Medical Center General hospital
Yoshikazu Yamamoto Tenriyorozu Hospital
Michio Tanaka Kyoto University Graduate School of Medicine
Akihiko Nakaizumi Kyoto University Graduate School of Medicine
- 2 -

Sponsors and supporters
Abbott Japan Co., Ltd.
ARKRAY Inc.
Beckman-Coulter Inc.
Denka-Seiken Co., Ltd.
Hitachi High-Technologies Corp.
Japan Association of Clinical Reagents Industries (JACR)
Japanese Society of Laboratory Medicine (JSLM)
Roche Diagnostics K.K.
Sekisui Medical Co., Ltd.
Siemens Japan K.K.
Sysmex Corp.
General Information
Dates
Thursday, 29 November– Saturday; 1 December, 2012
Venue
Kyoto International Conference Center (http://www.icckyoto.or.jp/en/index.html)
Room D
Official Language
English: No simultaneous interpretation will be provided for any language.
Tourist Information
Climate
The average temperature in Kyoto at the time of the meeting is 4.7 ~ 13.7°C.
Tipping and Consumption Tax
Tipping is not customary in Japan and is unnecessary in all situations. However, major restaurants or
hotels may automatically add a 10% to 15% service charge to your bill. A 5% consumption tax is also
automatically added to purchases at shops, restaurants and hotels.
Currency
Japanese yen (JPY) is the only currency accepted at regular stores and restaurants. Most currencies and
traveler’s checks can be exchanged at international airports, large branches of major banks, and hotels.
Banks are open from Monday to Friday, 9:00-15:00. The organizing committee will accept only
Japanese yen (JPY) is the only currency accepted at regular stores and restaurants. Most currencies and
traveler's checks can be exchanged at international airports, large branches of major banks, and hotels.
Banks are open from Monday to Friday, 9:00-15:00. The organizing committee will accept only
Japanese yen at the registration desk.
- 3 -

Electricity
The electric current in use is 100V 60Hz in Western Japan.
Insurance
The Organizing Committee can accept no responsibility for accidents or damage to the private property
of participants. Please make your own arrangements for health insurance, traveler's insurance, and any
other necessary insurance.
Passport and Visas
A valid passport is required to enter Japan. In addition, a visa is required for visitors from some nations;
please contact the Japanese Consulate or diplomatic mission in your country as soon as possible so that
those who are required to do so can take the necessary steps. For details, please refer to
http://www.mofa.go.jp/j_info/visit/visa/index.html (Guide to Japanese VISAS from the Ministry of
Foreign Affairs). If you are required to obtain a visa and some documents for visa purposes, please
contact the secretariat: ascpalm2006@congre.co.jp. Please note that this procedure is intended to assist
registered participants who need to obtain a visa and it does not imply any financial support from the
Meeting.
Time
Japan Standard Time is 9 hours ahead of Greenwich Mean Time.
From Airport to Kyoto International conference Center
- 4 -

Approximate Flight Times to Japan
From Asia
Bangkok [ 5hrs ] /Beijing [ 1.5hrs ] / Busan [ 1hr ] / Hong Kong [ 3hrs ] / Kuala Lumpur [ 6hrs ] /
Manila [ 4hrs ]/ New Delhi [ 8hrs ] / Seoul [ 1.5hrs ] / Ulaanbaatar{8hrs}/ Shanghai [ 1.5hrs ] /
Singapore [ 6hrs ] / Taipei [ 2hrs ]
The modern Kyoto Station serves as a link between Kyoto and the rest of the country.
- 5 -

Kyoto Heian Hotel(京都平安ホテル)
http://kyoto-heian-hotel.com/
Rubino Kyoto Horikawa Hotel
(ルビノ京都堀川ホテル)
http://www.rubino.gr.jp/
Kansai International Airport
- 6 -
Kyoto International Conference Center
http://www.icckyoto.or.jp/
Kyoto Station

The Kyoto City Subway system is easy to navigate, and is convenient for commuting from hotel to the
Conference Center.
- 7 -

A fully covered wheelchair-friendly tunnel and walkway stretches from the subway to the Conference
Center's front doors.
Floor Map (Room D)
Subway
Station
Entrance
Registration
- 8 -
Event Hall
Room D
Room H
Speaker
Preparation
Space

Scientific Program Information
Speaker preparation space
The speaker preparation space is set in room H near room D.
Opening hours;
Friday 30 November 08:00 – 17:00
Saturday 1 December 08:00 – 16:00
All oral presenters are asked to load / check their presentation at least one (1) hour prior to
their session commencing to ensure the presentation is checked and tested. You will be briefed
on how to use the system when you meet with the audio visual technicians.
Speakers for the featured oral session are asked to present their papers for less than 15 minutes,
leaving more than 5 minutes for discussion.
Information and Guidelines for Poster Presenters
Presentation
Poster boards will be located inside the Exhibition hall (Event Hall).
Poster presenters will be handed a ‘poster number’ upon registration, and must mount their
poster on their allocated board on the appropriate day as follows.
Poster #01 - #51
Mounting: 08:00 – 10:00 (Friday)
Removal: 17:00 – 18:00 (Friday)
Poster #52 - #99
Mounting: 08:00 – 10:00 (Saturday)
Removal: 18:00 – (Saturday)
The organisers are not responsible for any item, including posters, left in the room outside of
the allocated hours.
Poster presenters are asked to present a short oral presentation for less than 5 minutes and free
discussion for 5 minutes standing by their posters during the formal Poster Session (13:00 –
14:00 on Friday or 17:00 – 18:00 on Saturday) to field questions and discussion.
Display Facilities
� One panel is available for display of each poster. To fit within the poster frame,posters must be
sized as follows: posters must not exceed 90cm wide by 190cm high.
� A limited supply of Velcro for attaching the poster display to the fabric panel backing is supplied.
� Electrical outlets will not be provided in the poster presentation area.
- 9 -

Preparation of a Poster
� The official language for the posters is English.
� Prepare the poster on material that is lightweight. The material can be on one sheet so that it can be
rolled up for easy transport or on separate panels for individual mounting. Posters should be
readable from a distance of 2 meters. For adequate visibility,capital letters should be at least 1 cm
high after enlargement to full poster size.
� Your poster should be self-explanatory so that you are free to supplement and discuss particular
points raised by enquiry. It may include:
Diagrams and charts
Reaction schemes
Photographs
Example of Layout
Poster Layout
On the top left corner of your poster reserve a space 20cm wide x 20 cm high for the poster number
provided by the ASCPaLM organizer. This number will also be identified in the program, so people that
have an interest in your poster can easily find it.
20 cm
Aim
Title / Names / Institution
Materials and Methods
Results
Conclusions
70 cm
90 cm
- 10 -
20 cm
190 cm

Event Hall is located by the conference center at the
north side. A fully covered walkway stretches from the
conference room to the Event Hall.
- 11 -
Event Hall
Poster Session
Conference Room

Program at a glance
Nov. 29 (Thu) Nov. 30 (Fri) Dec. 1 (Sat)
8:50 Opening
9:00 Featured Oral Session
6 presentations
11:00 Innovative Molecular
Biomarkers in thrombus
Formation
12:00
-
12:55
13:00
(Sekisui Medical)
Luncheon seminar
The clinical significance and
measuring method of
albuminuria
T. Konta
(Siemens Healthcare)
Registration Poster session #1
(PO. 1 – PO. 51)
- 12 -
9:00 The Globalization of Medicine:
Appropriate Utilization of
Laboratory Tests
Keynote Lecture by
E. Blair Holladay
Executive Vice President/Chief Executive,
American Society for Clinical Pathology
10:00 Establishment of Reference
Intervals: Still a Challenging
Issue!
(Beckman Coulter)
12:00
-
12:55
13:00
-
14:50
Luncheon seminar
Small Dense LDL-Cholesterol
and Risk for CHD
Ron C. Hoogeveen
(Denka Seiken)
From diagnosis to personalized
medicine: comprehensive
approaches for patient care
14:00 Exhibition: site visit
(Roche Diagnostics)
15:00 National The Diagnosis and Treatment 15:00 Scientific Innovation Seminar
16:00
Representative
Meeting
of Leukemia: A look into the
future
-
16:50
‘Infectious Diseases’
(Abbott Diagnostics)
17:00 Welcome
Reception
(Sysmex Corp.)
17:00 Poster session #2
(PO. 52 – PO. 99) )
18:00 Banquet
18:00 Closing
20:00 (by Sysmex)
19:30 Farewell Party
(at Rubino-Horikawa Hotel)

Keynote Lecture
(Chaired by Mitsuru Murata)
The Globalization of Medicine: Appropriate Utilization of
Laboratory Tests
E. Blair Holladay, BA, BS, MS, Ph.D., SCT (ASCP) CM
Executive Vice President/Chief Executive
American Society for Clinical Pathology
As the borders between individual countries within the world edge “closer together” as a relationship
of travel, virtual communication (virtual mail, virtual conferences, social media) and ready access to the
internet and telepathology, healthcare providers are finding themselves regularly exchanging best
practices as well as discussing the latest research and disease management paradigms. As medical
laboratory practitioners, it is incumbent that we share best practices and also discuss which types of tests
are appropriate or inappropriate to conduct, depending on the clinical scenario. Pathologists and
laboratory professional are in a best position to help reduce healthcare expenditures by advising
clinicians and other healthcare providers about appropriate testing. The axiom, “right test, right patient,
right time and the right cost” is becoming more critical in an age of limited healthcare spending.
Examples of inappropriate and over utilized tests are pervasive throughout both anatomic and clinical
pathology and laboratory medicine, such as use of low-risk human papillomavirus testing for cervical
cancer screening3 (proven to have no relationship to the development of cervical cancer), and the
practice of unnecessary preoperative testing panels unrelated to the corresponding surgery (antinuclear
antibodies testing for lupus and rheumatoid, hepatitis A and B, etc.). Another factor is the poorly
characterized definitions of test use. Clinicians are often unclear about when or how to use many tests
and thus may not make the appropriate choices. With approximately 4,000 new tests on the horizon over
the next decade, it will be much more difficult for clinical to be kept informed on the evidenced-based
data that helps them to choose the most appropriate test, especially in the new and impending age of
genomics. Unnecessary testing can also result from duplicate orders and rework, such as when a
patient is transferred between heath settings without sharing of test data or when more than one
physician is involved in a patient’s treatment. Research indicates there is no connection between the
amount of laboratory testing done and the quality of care. Excessive testing may also lead to a chance of
false positive diagnoses. Approximately 5 percent of healthy patients get abnormal test results leading
to unnecessary testing and potentially risky treatments.
- 13 -

Featured Oral Presentation
Chairpersons: Won-Ki Min and Yukio Ozaki
1. CLINICAL PATHOLOGY SCOPE IN CANCER SURVEILLANCE
Siti Boedina Kresno
Dharmais National Cancer Center, Jakarta Indonesia
2. CLINICAL APLICATION OF LIQUID CHROMATOGRAPHY TANDEM MASS
SPECTROMETRY
Bai Hsiun Chen M.D.; FCAP
Department of Laboratory Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan,
Director, Division of Toxicology, Department of Laboratory Medicine, Kaohsiung Medical
University Hospital, Kaohsiung, Taiwan
3. SUSTAINED EXPRESSION OF A NEURON-SPECIFIC ISOFORM OF THE TAF1 GENE IN
DEVELOPMENT STAGES AND AGING IN MICE
Jamiyansuren Jambaldorj 1,2 , Satoshi Makino 2,3 , Gen Tamiya 2 , Batmunkh Munkhbat 1
1. Central Scientific Research Laboratory, Institute of Medical Sciences, Ulaanbaatar, Mongolia
2. Advanced Molecular Epidemiology Research Institute, Yamagata University Faculty of
Medicine, Yamagata, Japan
3. Molecular Neuroscience Research Center, Shiga University of Medical Science, Otsu, Japan
4. OPTIMIZATION OF PRONASE TREATMENT FOR X-MATCH TESTING
Gansuvd Balgansuren, MD, PhD 1 , Prof. Namid Munkhtuvshin MD,PhD 2
1
Clinical Transplantation Immunology Laboratory, Duke University Health System, Duke
University, 2 Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
5. INDIAN STUDY OF LIVER FUNCTION TESTS IN POST LIVER TRANSPLANT PATIENTS
Pradeep Naik, Premsagar.Bandi, Mallikarjuna Madhavarapu
Department of Clinical Biochemistry, Global Hospitals, Lakdikapool, Hyderabad-500004 INDIA
6. The Analysis on the familial tendency of Transferrin Saturation (TS) in Korean
Families Based on The Fifth Korea National Health and Nutrition Examination
Survey (KNHNES, V-1) 2010
Sung-Hee Oh, So-Young Kim, Woochang Lee, Sail Chun, and Won-Ki Min*
Department of Laboratory Medicine, Asan Medical Center and University of Ulsan College of
Medicine, Korea
- 14 -

FO 1
CLINICAL PATHOLOGY SCOPE IN CANCER SURVEILLANCE
Siti Boedina Kresno
Dharmais National Cancer Center,
Jakarta Indonesia
The development of molecular biology gives us a deeper understanding of the mechanism and
pathogenesis of disease and the medical science in the 21 st century will therefore be based on the
understanding of disease process at molecular levels. These new trends in the field of oncology
challenges clinical laboratory professionals to provide molecular diagnostic tools.
Cancer surveillance considered for this presentation is not monitoring of cancer trends over time or
collecting and analyzing data on cancer occurrence and risk factors at population levels, but in the
context of testing people who are at increased risk for developing cancer or have previously had cancer
to predict early recurrence or metastasis.
The scope of clinical pathology in surveillance, prediction and prognostication in cancer includes
hereditary cancer as well as sporadic cancer in particular by using lymphocytes for risk assessment of
hereditary cancer and using circulating tumor cells (CTC) and or circulating or cell-free DNA/RNA in
the prognostication of sporadic cancer.
An Indonesian experience in this field will be presented.
- 15 -

FO 2
CLINICAL APLICATION OF LIQUID CHROMATOGRAPHY TANDEM MASS
SPECTROMETRY
Bai Hsiun Chen M.D.; FCAP
Department of Laboratory Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan
Director, Division of Toxicology, Department of Laboratory Medicine, Kaohsiung Medical
University Hospital, Kaohsiung, Taiwan
Liquid chromatography-tandem mass spectrometry (LC-MS/MS) has seen enormous growth in
clinical laboratories during the last 10–15 years. It offers better analytical specificity than that of
immunoassays or conventional high performance liquid chromatography (HPLC) for low molecular
weight analytes and has higher throughput than gas chromatography-mass spectrometry (GC-MS)
which the analyte needs to be volatile and is more time consuming by derivatization. The main
applications of LC-MS/MS have been seen in the field of Drug/Toxicology and Biochemical
Genetics/Newborn Screening, but in recent years there had more application in the field of
Endocrine laboratories.
Measures to improve specificity and sensitivity include sample clean-up and optimising
chromatography to avoid interferences and ion suppression due to sample-matrix components.
We will discussed our experience of LC-MS/MS in developing quantification of arecholine,
melamine, ketamine, DEHP, 25(OH)D2 and 25(OH)D3.
- 16 -

FO 3
SUSTAINED EXPRESSION OF A NEURON-SPECIFIC ISOFORM OF THE
TAF1 GENE IN DEVELOPMENT STAGES AND AGING IN MICE
Jamiyansuren Jambaldorj 1,2 , Satoshi Makino 2,3 , Gen Tamiya 2 , Batmunkh Munkhbat 1
1. Central Scientific Research Laboratory, Institute of Medical Sciences, Ulaanbaatar, Mongolia
2. Advanced Molecular Epidemiology Research Institute, Yamagata University Faculty of Medicine,
Yamagata, Japan
3. Molecular Neuroscience Research Center, Shiga University of Medical Science, Otsu, Japan
TATA-box binding protein associated factor 1 (TAF1) protein is the largest and the essential component
of the TFIID complex in the pathway of RNA polymerase II–mediated gene transcription, and it
regulates transcription of a large number of genes related to cell division. The neuron-specific isoform
of theTAF1 gene (N-TAF1), which we reported previously, may have an essential role in neurons
through transcriptional regulation of many neuron-specific genes. In the present study, we cloned the
full-length cDNA that encodes the mouse homologue of N-TAF1 (N-Taf1) protein. By carrying out of
real time RT-PCR, we investigated the expression analysis of the N-Taf1 mRNA in mouse tissues and
cell lines. As well as the human N-TAF1, the N-Taf1 showed limited expression in the brain and
neuroblastoma, whereas Taf1 expressed elsewhere. Furthermore, in mouse embryo head or mouse brain,
mRNA expression of TAF1 changes dramatically during development but N-Taf1 showed sustained
expression. Our result suggests that the N-Taf1 gene has an important role in non-dividing neuronal cell
rather than in cell division and proliferation during neurogenesis.
- 17 -

FO 4
OPTIMIZATION OF PRONASE TREATMENT FOR X-MATCH TESTING
Gansuvd Balgansuren, MD, PhD 1 , Prof. Namid Munkhtuvshin MD,PhD 2
1 Clinical Transplantation Immunology Laboratory, Duke University Health System, Duke University,
2 Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
Objectives: Due to increased receipt of crossmatch test requests from patients treated with rituximab
desensitization therapy we aimed to optimize pronase concentration for routine testing that could
significantly reduce non-specific background binding CD20 while keeping CD19.
Materials and Methods: Peripheral blood mononuclear cells (PBMNC) were incubated with different
concentrations (0.5; 1.0; 1.5 and 20 mg/ml) of pronase for 30 minutes at 37°C. Two kinds of normal
human sera (GEN & GTI) and positive pooled serum (PPS) were used as negative and positive controls.
10 µg/ml rituximab spiked NHS and rituximab treated patient’s sera were used as test samples.
Anti-human CD19 and CD20 antibodies were used for phenotypic analysis.
Results: 15 min incubation of donor cells with various concentrations of pronase resulted no change on
CD20 expression, however extended incubation of donor cells with pronase at the dose of 1.5 mg/ml
and 2.0 mg/ml for 30 min completely removed CD20 from B cell surface. The latter dose also markedly
reduced CD19 expression (55%±26%, n=8) as well as its intensity. Therefore, 1.5mg/ml of pronase has
been chosen for further X-Match testing. MCS of rituximab spiked NHS (GEN & GTI) with
un-pronased donor cells in B cell X-Match was 512±50 & 452±53 and which has reduced up to 68±28
& 47±18 with 1.5 mg/ml pronase treated donor cells, respectively (n=20). Rituximab treated patients
sera with un-pronased donor cells in B cell X-Match showed MCS of 445, 447 and 468 which has also
reduced up to 20, 48 and 6 with 1.5mg/ml pronase treated donor cells. The pronase treatment
significantly reduced non-specific background binding in both T and B cell X-Match and significantly
increased sensitivity only in T cell X-Match.
Conclusion: 1.5 mg/ml pronase treatment of donor cells for 30 min at 37°C could completely remove
CD20 from B cell surface while keeping sufficient number of CD19 positive cells to safely X-Match
rituximab treated patients alone with other patients.
- 18 -

FO 5
INDIAN STUDY OF LIVER FUNCTION TESTS IN POST LIVER TRANSPLANT
PATIENTS
Pradeep Naik, Premsagar.Bandi, Mallikarjuna Madhavarapu
Department of Clinical Biochemistry, Global Hospitals, Lakdikapool, Hyderabad-500004 INDIA
Liver transplantation means surgical replacement of a diseased liver with a healthy liver. The survival
rate was 30% after one year. LTx was considered to be the last procedure when all medical or surgical
intervention failed. Advances in donor organ preservation, surgical techniques, patient selection,
immunosuppressive regimens and treatments for opportunistic infections all have contributed to
substantially improve the survival rates. Despite substantial technological, medical and surgical
advances, liver transplantation remains a complex procedure that is accompanied by significant
morbidity and mortality.
The post-operative outcome of each patient varies greatly depending on the patient’s pre- operative state,
quality of the donated organ and the complexity of the surgery. Complications occur both immediately
post transplant and in the long term. Most of the problems can be satisfactorily assessed with a panel of
routine LFTs results of which are generated quickly, cheaply on the analyzer which operates 24 hrs.
Liver Function Test identifies the presence of problem but not problem itself. Abnormal results can be
meaningful only when used with clinical data, radiological findings.
The study includes 75 post LTx patients in three groups Adults (Non ACR ), Pediatrics and ACR. All
recipients were on immunosuppressive therapy (tacrolimus, micophenolate and Methylprednisolone),
antiviral (gancyclovir), antiprotozoal, antibacterial and antifungal (fluconazole). 5 ml of blood was
drawn in plain vacutainer from the post LTx patients every day for 15 days and LFT and GGT was done.
Routinely performed liver function tests correlates well with clinical complications involving liver in the
transplant patients. Instead of daily testing, may be alternate day analysis of LFT should be sufficient for
effective monitoring of patients. The total protein and albumin and the transaminases offer little help in
monitoring LFT post LTx.
The elevated levels of serum GGT and ALP may be related to chronic immune damage to the
transplanted liver. Serum GGT and ALP can be used as early markers for diagnosing biliary
complications and can be used to asses adequacy of endoscopic treatment in the group of patients
presenting early. Thus, most of the problems can be satisfactorily assed with a panel of routine LFTs
generated quickly, cheaply on analyzer which operates 24 hrs each day. However, it must be emphasized
that LFTs may identify the presence of problems but not the problem itself and the abnormal results are
meaningful only when correlated with other clinical information.
- 19 -

FO 6
The Analysis on the familial tendency of Transferrin Saturation (TS) in Korean
Families Based on The Fifth Korea National Health and Nutrition Examination
Survey (KNHNES, V-1) 2010
Sung-Hee Oh, So-Young Kim, Woochang Lee, Sail Chun, and Won-Ki Min*
Department of Laboratory Medicine, Asan Medical Center and University of Ulsan College of Medicine,
Korea
Background TS is an index used to screen hereditary hemochromatosis (HH), a disease characterized
by the deposition of excessive amounts of iron in parenchymal cells. Despite very low incidences of
C282Y and H63D mutations, well-known common genetic causes of HH, some studies showed higher
TS level in Koreans compared with those of Caucasians. This study aims to compare TS level between
Korean population and the U.S. Caucasian by using the data of KNHANES V-1 and National Health and
Nutrition Examination Survey (NHANES), 2001-2002. In addition, we tried to assess the existence of a
familial tendency in Korean’s TS level by statistical analysis of parents’ and children’s TS value.
Methods Among 8958 KNHANES V-1 subjects, those without anemia and hepatitis B antigen were
enrolled in this study. The age distribution of 2946 males was 10-91 years old (mean±SD, 43.3±19.3)
and 10-93 years old for 3243 females (44.6±18.6). 557 families with at least one child were included for
an analysis for familial tendency and total number of subjects within them was 1907.Each parent was
grouped into four quartiles (designated as 1, 2, 3 and 4) based on his or her TS levels and then the mean
of paternal and maternal TS four quartile indices (MTQI) were calculated and used for further analysis.
Their offspring’s mean TS levels (OMTL) among each MTQI group were compared using one-way
analysis of variance and post-hoc multiple comparison to show the statistical differences. NHANES
2001-2002 was used to evaluate the mean TS level in Caucasian and unpaired student t-test was
performed to assess the statistical difference. We considered a p-value

Advanced Technical Seminar
Sekisui Medical Co., Ltd.
Innovative Molecular Biomarkers in thrombus Formation
Siemens Japan K.K.
The clinical significance and measuring method of
albuminuria
Sysmex Corporation
The Diagnosis and Treatment of Leukemia: A look into the
future
Beckman-Coulter Inc.
Establishment of Reference Intervals: Still a Challenging
Issue!
Denka-Seiken Co., Ltd.
Small Dense LDL-Cholesterol and Risk for CHD
Roche Diagnostics K.K.
From diagnosis to personalized medicine: comprehensive
approaches for patient care
Abbott Japan Co., Ltd.
Scientific Innovation Seminar
‘Infectious Diseases’
- 21 -

Innovative Molecular Biomarkers in thrombus
Formation (Sekisui Medical)
FDP and D-dimer: How we use them in various clinical situations
Mitsuhiro Uchiba
Department of Blood Transfusion and Cell Therapy, Kumamoto University Hospital Chuoku Honjo
1-1-1 Kumamoto, Japan
Fibrin degradation product (FDP) and D-dimer are important molecular marker of coagulation and
fibrinolysis, and are generated from fibrin by plasmin. Thrombin, an end product of coagulation cascade,
releases fibrinopeptide from fibrinogen and thereby produces fibrin monomer. Fibrin monomers
aggregate each other, and finally form unstable fibrin. After generation of unstable fibrin, D-domain of
fibrin molecules are covalently cross-linked each other by coagulation factor XIII. Finally, stable fibrin
is generated. Plasmin potentially degrades any types of fibrin described above. As D-dimer is generated
only from cross-linked fibrin, increasing D-dimers indicates presence of stable fibrin and its degradation
by plasmin, and is useful for diagnosis of deep vein thrombosis (DVT). However, D-dimers are
generated from any types of stable fibrin including fibrin in hemostasis thrombi on wound and in ascites.
Therefore, D-dimers are used as rule out marker for DVT. In contrast, FDPs are generated any type of
fibrin including fibrin monomer and unstable fibrin. Therefore, FDP sensitively responses in fibrin
formation and fibrinolysis. Also FDP increases early phase of activation of coagulation without or
slightly increase in D-dimer. FDP is a useful marker for diagnosis of DIC. FDP, but not D-dimer, is
potentially produced from fibrinogen. FDP increases in fibrinogenolytic condition resulting uncontrolled
fibrinolysis situation.
- 22 -

Measurement of Albuminuria (Siemens
Healthcare)
The clinical significance and measuring method of albuminuria
Tsuneo Konta
Department of Cardiology, Pulmonology, and Nephrology, Yamagata University School of Medicine
In Japan, a number of dialysis patient keeps rising continuously, reaching more than 300,000 this year.
Although diabetic renal disease alone accounts for more than 40% of the total primary disease for
dialysis, the early detection and prompt therapy can suppress the progress of the disease.
The early stage of diabetic nephropathy does not cause any certain symptoms except moderate increase
of urinary albumin excretion (“microalbuminuria”). If untreated, it can progress into macroalbuminuria,
irreversibly damage the kidney and eventually lead into end-stage renal failure. Thus, the early detection
of nephropathy by monitoring urinary albumin excretion is essential to treat diabetic patients.
In recent years, microalbuminuria is known to exist in the majority of hypertensive patients, besides
diabetic patients. Furthermore, the presence of microalbuminuria is closely correlated with the incident
of cardiovascular diseases; and the decrease of urinary albumin excretion can abate these risks.
However, urinary albumin is not measured adequately, even for diabetes patients, because it takes time
to analyze and the frequency of its measurement is limited in Japan. Moreover, the urinary albumin
concentration can be affected by body position and fluid intake. To diagnose albuminuria correctly, it is
recommended to measure both albumin and creatinine levels in morning urine sample.
In June 2012, a new CKD treatment guide 2012 is launched by the Japanese Society of Nephrology,
recommending the use of urinary albumin/creatinine ratio for diabetic patients and urinary
protein/creatinine ratio for nondiabetic patients for CKD classification. In both cases, the correction by
urinary creatinine is essential to alleviate the urinary concentration.
At this seminar, the clinical data will be presented, including the association between urinary albumin,
kidney disease, and cardiovascular disease, and the importance of creatinine modification for the
accurate evaluation of urinary albumin excretion; based on Japanese patients.
- 23 -

The Diagnosis and Treatment of Leukemia: A
look into the future(Sysmex)
1. The Current Status of the Diagnosis and Classification of Hematopoietic Malignancies
A . Aziz Baba
Stem Cell Transplantation and Clinical Haematology Unit, School of Medical Sciences, Universiti Sains
Malaysia, USM-Health Campus, 16150 Kubang Kerian Kelantan, Malaysia
2. Challenges of Cytogenetics &FISH Diagnosis in Leukaemias and Lymphomas
Tien Sim Leng
Department of Haematology and SGH Blood Bank & Cytogenetics Department of Pathology Singapore
General Hospital, Outram Road, Singapore 169608
3. The Latest Topic of Stem Cell Transplantation for Leukemia and its Development in the Future in
Taiwan
Jih-Luh Tang
TaiCheng Stem Cell Therapy Center, National Taiwan University Hospital, 10002 No.7,
ZhongShan South Road, Taipei, Taiwan
- 24 -

The current status of the diagnosis and classification of hematopoietic malignancies
A . Aziz Baba
Stem Cell Transplantation and Clinical Haematology Unit, School of Medical Sciences, Universiti Sains
Malaysia, USM-Health Campus, 16150 Kubang Kerian Kelantan, Malaysia
The hematopoietic malignancies are a diverse group of disease entities that include the lymphomas,
leukemias, myeloproliferative neoplasms, myelodysplastic syndromes, plasma cell dyscrasias,
histiocytic and mast cell neoplasms. Over the years multiple classification systems have been employed
to provide reliable criteria in their diagnosis and their classification into clinically relevant disease
entities. Earlier systems were predominantly based on cellular morphology and tissue architecture.
Increased knowledge and understanding of the genetic and molecular basis of these disorders has led to
continuous reassessment and refinement of the classification with incorporation of cytogenetic and
molecular genetic analyses as part of the diagnostic criteria. In this talk I will discuss changes and
issues regarding the revised World Health Organisation (WHO) classification of hematopoietic and
lymphoid neoplasms. The integration of molecular information together with morphology,
immunophenotype and clinical information has helped to further refine the WHO classification and
improve its prognostic value. Illustrative examples, with an emphasis on the myeloid neoplasms will
be presented.
- 25 -

Challenges of Cytogenetics &FISH Diagnosis in Leukaemias and Lymphomas
Tien Sim Leng
Department of Haematology and SGH Blood Bank & Cytogenetics Department of Pathology Singapore
General Hospital, Outram Road, Singapore 169608
All human DNAs including the genes are neatly packed into 46 chromosomes, 22 pairs of autosomal
chromosomes and one pair of sex chromosomes. Cytogenetics is a branch of genetics that is concerned
with the study of chromosomes. When combined with molecular techniques such as Fluorescence in-situ
Hybridization (FISH), cytogenetic analysis has widespread clinical applications in the diagnosis of
leukaemias and lymphomas.
Cytogenetic studies and FISH are now offered as a standard of care in the diagnosis of patients with
leukaemia and lymphoma. However, there are challenges with the use of these two diagnostic tools and
their limitations will be discussed. Improved processes and techniques including slide dryer chamber
and metaphase finder are employed to enhance the efficiency and to improve turn-around times.
In leukaemias and lymphomas, cytogenetic study plays a significant role in the detection of abnormal
neoplastic clones, classification of malignant disorders, prediction of disease progression, monitoring
response to treatment, detection of minimal residual disease and institution of targeted therapy, notably
chronic myeloid leukaemia (CML). Molecular targeted drugs and the treatment outcome in CML will be
illustrated. Various recurring chromosomal aberrations in acute and chronic leukaemias as well as
lymphomas have been found. In bone marrow or stem cell transplantation, cytogenetic is used to
determine engraftment or chimerism and to assess disease remission or relapse. FISH strategies using
various fluorescent-labeled DNA probes including FISH panel testing enhance the usefulness of
cytogenetic analysis of leukaemias and lymphomas. Other strategies include comparative genomic
hybridization (CGH) arrays and multi-colour FISH (M-FISH).
Any new discovery of more specific genes and recurring chromosomal aberrations in leukaemias and
lymphomas in conjunction with the availability of targeted FISH probes will enhance the diagnosis of
leukaemias and lymphomas which may lead to better risk stratification and more concise molecular
targeted therapy in future.
- 26 -

Establishment of Reference Intervals: Still a
Challenging Issue! (Beckman Coulter)
Kiyoshi Ichihara and Jeong-Ho Kim (Chairpersons)
1) What is an optimal protocol for the IFCC global study on reference values?
Kiyoshi Ichihara, MD & PhD (Japan)
2) Controversies over statistical methods for derivation of reference intervals.
Swarup A Shah, PhD, and Tester F Ashavaid, PhD (India)
3) Analytical issues in conducting the multicenter studies on reference values.
Kiyohide Ishikura, PhD (Japan)
4) Age-specific reference intervals: derivation for pediatric population from the hospital information
system.
Jeong-Ho Kim, MD & PhD (Republic of Korea)
Although global effort led to standardization of many of the common laboratory tests, their reference
intervals (RI) remain widely variable amongst laboratories. This reflects difficulty in deriving RIs by
individual laboratories and highlights the necessity of deriving it collaboratively for common use. The
guidelines used for defining, establishing and verifying RIs are available, which was published by CLSI
in collaboration with IFCC. There is a strong global need for establishment of proper RIs for common
use by conducting multicenter studies. However, there remain controversies on various aspects of
conducting the study.
In this symposium, Prof. Ichihara presents the common protocol for the global multicenter study on
RIs, which is proposed as well as currently being conducted by Committee on Reference Intervals and
Decision Limits (C-RIDL) of IFCC. In order to validate the protocol, the following aspects will be
discussed: 1) criteria for recruitment of healthy volunteers, 2) ways of controlling pre-analytical
variables, 3) harmonization of results across collaborating countries based on common measurements of
a panel of sera, 4) methods for secondary (posteriori) exclusion based on test results, 5) transference of
the established RIs based on a “mini panel” of sera.
Dr. Ishikura, a member of C-RIDL, will present an overview about the feasibility of the panel-of-sera
strategy in harmonizing results worldwide, and will also discuss the possible use of mini-panel in the
transference of established RIs.
Dr. Shah and Dr. Ashavaid will talk on the controversies over the use of parametric or nonparametric
computation of RIs and secondary exclusion of individuals after obtaining testing results. The issues
based on the simulation study will also be discussed.
Prof. Kim will talk about the difficulty in establishing RIs for the pediatric, and geriatric population.
He will review the issue in the literature and will present his approach in selecting appropriate reference
individuals from large hospital records for deriving the pediatric RIs,
With this overview, we would like to rationally discuss the optimal ways of establishing the RIs at the
- 27 -

end of the session.
Establishing pediatric reference interval for 13 clinical chemistry tests derived from
normal subjects in the pediatric endocrinology clinic in Korea
Jeong-Ho Kim 1) , Sun-Mi Cho 1) , Youkyung Seo 1) , Ho Seong Kim 2) , Woonhyoung Lee 1) , Oh Hun
Kwon 1) , *
Departments of Laboratory Medicine 1) and Pediatrics 2) , Severance hospital, Yonsei
University College of Medicine, Korea
Background: Defining pediatric reference intervals is one of the most difficult tasks for laboratory
physicians. The continuously changing physiology of growing children makes their laboratory values
moving targets. In addition, ethnic and behavioral differences may also cause variations. Age- and sexspecific
partitioned reference intervals of 13 serum biochemical analytes in Korean children were
presented.
Methods: A total of 2,474 patients, girls from 2 to 14 years and boys from 2 to 16 years, tested in a
short stature workup but diagnosed as normal at the pediatric endocrinology clinic of Severance
Hospital from Sep 2010 to Jun 2012 were included in this study. Calcium, inorganic phosphorus, blood
urea nitrogen (BUN), creatinine, uric acid, glucose, total cholesterol, total protein, albumin, alkaline
phosphatase (ALP), aspartic aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin
test were performed on Hitachi 7600 analyzer (Hitachi High-Technologies, Tokyo, Japan). Partitionings
for gender or age group were performed by ANOVA, Mann Whitney test, Harris and Boyd method
(Clin Chem 1990;36:265-70), and Lahti criteria (Clin Chem 2004;50:891-900).
Results: Age specific partitioned reference intervals for ALP, creatinine, and total bilirubin in both
genders could be established. In addition, age specific partitioning of AST in female and total protein
and uric acid in male was required. However, age or gender partitioning of calcium and albumin in
children was not required.
Conclusions: Age and gender specific partitioned pediatric reference intervals of 13 clinical chemistry
assays derived from Korean healthy children performed in pediatric endocrinology clinic could be
established.
- 28 -

Innovative Lipid Biomarkers (Denka Seiken Co.)
Small Dense LDL-Cholesterol and Risk for Coronary Heart Disease: Recent
Findings from the Atherosclerosis Risk in Communities (ARIC) Study
Ron C. Hoogeveen, Ph.D., Assistant Professor
Section of Atherosclerosis and Vascular Medicine, Department of Medicine, Baylor College of Medicine
and Center for Cardiovascular Disease Prevention, Methodist DeBakey Heart and Vascular Center,
Houston, TX
Background- Evidence from in vitro studies indicates that small dense LDL (sd-LDL) is more
atherogenic than large buoyant LDL (lb-LDL). Furthermore, circulating levels of sd-LDL are highly
correlated with triglycerides and are increased in individuals with an atherogenic lipoprotein profile
such as patients with diabetes and metabolic syndrome. Previously, sd-LDL has been associated with
risk for vascular disease. However, the lack of a standardized sd-LDL assay which can be performed in
routine clinical laboratories has hampered its clinical application.
Objectives- We recently examined the relationship between plasma sd-LDL-cholesterol (sd-LDL-C)
level and risk for incident coronary heart disease (CHD) and stroke in the ARIC cohort.
Methods- Plasma sd-LDL-C was measured in 11,419 men and women of the biracial ARIC study using
a newly developed automated homogeneous assay. A proportional hazards model was used to examine
the relationship between sd-LDL-C, vascular risk factors, and risk for CHD events and stroke over a
period of ≈10 years.
Results- Mean plasma sd-LDL-C was higher in Caucasians than in African Americans (45.2 vs. 37.4
mg/dL, p

From diagnosis to personalized medicine:
comprehensive approaches for patient care
(Roche Diagnostics)
Yasuhito Tanaka (Chairperson)
Professor,Director
Department of Virology & Liver Unit, Crinical Laboratory
Nagoya City University Graduate School of Medical Sciences
(Speakers)
Reinhard Bertele (Roche Diagnostics K.K.)
Chris Newhouse (Roche Diagnostics K.K.)
Shigehisa Nagahashi (Roche Diagnostics K.K.)
The field of diagnostics is evolving from simply providing tools for disease diagnosis, to also providing
tools for personalized medicine. Roche has an industry leading portfolio, which utilizes many
technologies to support disease diagnosis, monitoring, and therapy guidance as well as treatment options.
We would like share with you some insight into some of the new tools we have developed for
physicians to manage patient care in the areas of infectious diseases and oncology and which are
currently in focus in the medical community. This seminar will cover recent advances in the fields of
Immunochemistry, Nucleic Acid Testing, and Automated Tissue Diagnostics.
Roche has developed a comprehensive panel of immunoassays, coupled with nucleic acid tests which
cover a wide variety of applications such as screening, diagnosis of acute and chronic infection and
therapy monitoring. In addition, in the field of oncology, there are many new tools which analyze the
disease status within tissue samples. All of these examples bring us step-by-step closer to having truly
personalized medicines.
Many of the above tools come together as part of Roche’s leading efforts in the development of
diagnostic tests along with innovative drugs to realize the dream of personalized healthcare. Roche
continues to improve our tests and to expand the portfolio for different clinical settings, maintaining our
global leadership. Roche continues to invest in many exciting activities on our continuous path to
innovate healthcare.
- 30 -

Scientific Innovation Seminar (Abbott
Diagnostics)
1. Meeting the Challenge of HIV Diversity and Application of Metagenomic-Based Technologies
for Virus Discovery
John R Hackett, Jr., Ph.D.
Abbott Laboratories, 100 Abbott Park Road, D-09GN, Bldg. AP20, Abbott Park, IL 60064-3500
USA
2. PLEX-ID Technology as a Single Tool for the Detection and Typing of Microbiological
Infections in Immuno-Compromised Patients
Jérôme Le Goff 1 , Linda Feghoul 1 , Jean Menotti 1 , Jean Luc Donay 1 , Séverine Mercier-Delarue 1 ,
Jean Louis Pons 1 , Marcus Picard-Maureau² and François Simon 1
1- Université Paris-Diderot, Hôpital Saint-Louis, Paris, France
2- Abbott Ibis Biosciences, 65205 Wiesbaden, Germany
- 31 -

Meeting the Challenge of HIV Diversity and Application of Metagenomic-Based
Technologies for Virus Discovery
John R Hackett, Jr., Ph.D.
Abbott Laboratories, 100 Abbott Park Road, D-09GN, Bldg. AP20, Abbott Park, IL 60064-3500 USA
The high level of genetic diversity characteristic of Human Immunodeficiency Virus Type 1 (HIV-1) has
significant implications for diagnosis, screening, vaccine development, and patient monitoring.
Phylogenetic analysis has revealed four distinct lineages or groups of HIV-1, designated as groups M, N,
O and P. Group M strains are further subdivided into subtypes A-D, F-H, J and K. For env, sequence
differences between subtypes and groups of HIV-1 range from 15-25% to 50%, respectively.
Recombination greatly increases overall diversity. Currently, more than 51 circulating recombinant
forms and a myriad of unique recombinant forms have been identified; recombinant strains constitute
>20% of worldwide HIV-1 infections. Recognizing the implications of HIV-1 diversity, the Abbott
HIV-1 Global Surveillance Program was established. Goals of this comprehensive program include:
monitoring global diversification of HIV-1, identifying newly emerging variants, and development of
panels of genetically and geographically diverse specimens. Studies advancing our understanding of
HIV-1 diversity will be presented. Globalization, growing numbers of immunocompromised patients,
and environmental changes have fueled a need for techniques to detect novel or previously unrecognized
pathogens. Recent advances in metagenomic technologies have transformed virus discovery. The
UCSF-Abbott Viral Diagnostics and Discovery Center represents one of few laboratories in the world
that leverages both pan-viral DNA microarrays (Virochip) and high-throughput sequencing for viral
discovery and screening. Both technologies offer a unique opportunity to screen in a non-biased
manner for all known viruses as well as for unknown viruses. Recent advances in application of these
metagenomic tools will be discussed. Advancement of these technologies offers unprecedented
opportunities for pathogen discovery and diagnostics.
- 32 -

PLEX-ID Technology as a Single Tool for the Detection and Typing of Microbiological
Infections in Immuno-Compromised Patients
Jérôme Le Goff 1 , Linda Feghoul 1 , Jean Menotti 1 , Jean Luc Donay 1 , Séverine Mercier-Delarue 1 ,
Jean Louis Pons 1 , Marcus Picard-Maureau² and François Simon 1
1- Université Paris-Diderot, Hôpital Saint-Louis, Paris, France
2- Abbott Ibis Biosciences, 65205 Wiesbaden, Germany
Introduction : the PLEX-ID technology (Abbott Ibis Biosciences), combining broad-range multiwell
PCR and mass spectrometry enables the detection and identification of a large spectrum of pathogens in
a single test. The PLEX-ID analysis is based on a broad multiwell PCR followed by an electrospray
ionization of the generated amplicons to analyse their time of flight by mass spectrometry. The weight
of the amplicons allows to determine their base compositions. The PLEX-ID technology allows to
identify large a variety of pathogens from various sample types (blood, plasma, biopsy, CSF, BAL..) in a
few hours, enabling the resolution of mixtures and according to the assay, a high resolution in
genotyping. A large panel of assays is available, from public health and biothreat to medical diagnostics.
That integrated solution allows the analysis of up to 15 x 96-well plates over 12 hours for a total
throughput of up to 250 samples per day.
We evaluated in our university hospital, specialized in Hematopoietic stem cells (HSC) and kidney
transplants, the performances and reliability of the PLEX-ID technology.
Objectives : to compare the accuracy and the rate of infections detected by PLEX-ID clinical assays and
routine standard methods and to compare the turnaround times of both procedures.
Methods : We evaluated 3 different clinical PLEX-ID assays for medical microbiology.
- The Viral IC assay detects 174 RNA and DNA viruses species and 2611 subspecies. We first analyzed
275 frozen samples from 54 children and 11 adults infected with Adv (245) and from non-infected
patients (30). In a second study, we parallelized the routine vs PLEX-ID in 79 fresh samples from HSC
and kidney transplants.
- The BAC Detection assay identifies 6026 species of bacteria and Candida and 4 antibiotic
resistance markers ( kpc, mec A, van A, van B). Peripheral blood and catheter sampling were tested in
BACT Alert blood culture (bioMérieux) and PLEX-ID. Preliminary study included 48 samples from
adult patients.
- The Broad Fungal assay identifies 2108 species. Fresh broncho-alveolar lavages and nasopharyngal
aspirates from 25 patients were compared between fungal culture and PLEX-ID analysis.
Additional samples collected from patients with undiagnosed fever were included in the BAC and
Fungal studies.
Results : All the negative PLEX-ID results in clinical samples were negative in the routine procedure
considered as the gold standard, pointing the excellent PLEX-ID negative predictive value. A high
negative predictive value was also observed with stored plasma samples in Viral IC assay.
- Viral IC assay in Adenovirus-infected patients: ADV was detected in 96.4% of ADV positive stored
plasma. The accuracy for species identification was 0.985. Out of 29 routine-negative samples collected
from known ADV infected patients, 7 were positive with PLEX-ID. Viral loads determined with
- 33 -

Adenovirus R-gene were correlated with PLEX-ID levels up to 6 log10 copies/ml (r²=0.67, p

POSTER SESSION
1. Clinical Chemistry (PC 1-PC 21)
Midori Masuda (Chairperson)
No. 1. Tacrolimus in Adult Renal Transplant Recipients
Pradeep Naik, Mallikarjuna Madhavarapu
Department of Clinical Biochemistry, Global Hospitals, Lakdikapool, Hyderabad-500004, India
No. 2. Soluble FcγRIIIa Mφ levels in plasma correlate with maximum plaque diameter in patients
examined with carotid arterial echography
Midori Masuda 1 , Keiko Kouno 2 , Noriko Nishimura 1 , Hiroya Masaki 1 , Toyohiko Yokoi 1 , Masamichi
Yoshika 1 , Yutaka Komiyama 1 , Toshiji Iwasaka 2 , and Hakuo Takahashi 1
Departments of 1 Clinical Sciences and Laboratory Medicine, and 2 Medicine II, Kansai Medical
University, Osaka, Japan
No. 3. The Role of S100B Protein and GFAP in Predicting Discharged NIHSS of Anterior Circulation
Ischemic Stroke
Yenny Surjawan, Suryani As’ad, Teguh A.S Ranakusuma, Andi Wijaya
Medical Faculty of Hasanuddin University, Indonesia
No. 4. Detection of hereditary abnormality of phenylalanine metabolism using microbiologic method
E.Shuree 1 , G.Oyungerel 2 , N. Munkhtuvshin 2
1
Oyushur Laboratory, Ulaanbaatar, Mongolia
2
Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
Hitoshi Chiba (Chairperson)
No. 5. External quality assessment schemes for Clinical Chemistry in Mongolia
G.Oyungerel 1 , Ya Amarjargal 2 , N.Munkhtuvshin 1
1
Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
2
Dept. Medical Care Policy Implementation and Coordination, MOH, Mongolia
No. 6. Analysis of cholesteryl ester hydroperoxides in oxidized lipoprotein by Use of an Orbitrap
LC-MS
Shu-Ping Hui 1 , Toshihiro Sakurai 1 , Futaba Ohkawa 1 , Hiroaki Furumaki 1 , Shigeki Jin 1 , Hirotoshi
Fuda 1 , Seiji Takeda 1 , Takao Kurosawa 2 , Hitoshi Chiba 1
- 35 -

1 Faculty of Health Sciences, Hokkaido University, Kita-12, Nishi-5, Sapporo 060-0812, Japan 2
Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Ishikari-Tobetsu,
Hokkaido 061-0293, Japan
No. 7. An approach to chemical education at medical technologist training institutions in Japan
Hidetusgu Kohzaki
Department of Cell Biology, Institute for Virus Research, Kyoto University, Japan
No. 8. Development of the enzymatic method of serum ethanolamine and examination of its clinical
utility
E. Ohta (1) , E.Hokazono (1) , M.Ono (2) , T.Hotta (2) , S.Osawa (3) , Y.Kayamori (1)
1 Division of Medical Technology, Department of Health Sciences, Graduate school of Medicine,
Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka City, Japan
2 Department of Clinical Chemistry and Laboratory Medicine, Kyushu University Hospital, 3-1-1
Maidashi, Higashi-ku, Fukuoka City, Japan 3 Division of Clinical Laboratory Science, Department
of Medical Risk And Crisis Management, Chiba Institute of Science, 15-8 Shiomi-cho, Choshi,
Chiba, Japan
Masaharu Yamazaki (Chairperson)
No. 9. Establishment of enzyme-linked immunosorbent assay for urinary Tamm-Horsfall protein
Aki Nakayama 1 , Ryo Kubota 2 , Minoru Sakatsume 3 , Shiro Iijima 1,4 , Kiyoko Shiba 4
1. Faculty of Health Science Technology, Bunkyo Gakuin University, Japan
2. Department of Health Sciences, Saitama Prefectural University, Japan
3. Division of Clinical Nephrology and Rheumatology, Graduate School of Medical and Dental
Sciences, Niigata University, Japan, Graduate School of Health Care Science, Bunkyo Gakuin
University, Japan
No. 10. Direct analysis of chemical substances in biofluids without sample pretreatment
Ritsuko Wakayama 1 , Yasushi Iwasaki 1 , Motoaki Kamachi 1 , Natsumi Sawa 2 , Katsuko Hara 2 , Yutaka
Komiyama 2
1 Showa Denko K.K, Japan, 2 Department of Clinical Sciences and Laboratory Medicine, Kansai
Medical University Takii hospital, Japan
No. 11. Indoxyl Sulfate is an Independent Risk Factor for Coronary Heart Disease in Patients with
Chronic Kidney Disease
Ippei Watanabe 1 , Shunji Namba 2 , Yuko Okuda 2 , Akiko Morimoto 2 , Tetsumi Kojima 2 , Masanari
Sano 2 , Toshisuke Morita 1
1. Deartment of Laboratory Medicine Graduate School of Medicine Toho University, Japan
2. Clinical Laboratory Toho University Omori Medical Center, Japan
No. 12. Reference value for serum folic acid level in Japanese adults
- 36 -

Naoko Yamaguchi, Tomohiro Takeda, Chizuko Kuramoto, Takao Uchiike, Masaharu Yamazaki, and
Yasuyuki Okamoto
Central Clinical Laboratories, Nara Medical University, Japan
Konen Obayashi (Chairperson)
No. 13. Effects of alpha lipoic acid as an antioxidant in preventing renal mitochondrial oxidative
damage in diabetic rats
Noraihan Mat Harun, Rahajoe Imam Santosa
Department of Basic Medical Sciences, Kulliyyah of Medicine, International Islamic University,
Malaysia, Kuantan-Malaysia
No. 14. Detection of autoantibodies against ATTR in patients with FAP ATTR V30M
Konen Obayashi 1 , Masayoshi Tasaki 2 , Satoru Shinriki 1 , Genki Suenaga 2 and Yukio Ando 2
1
Diagnostic Unit for Amyloidosis, Department of Laboratory Medicine, Kumamoto University
Hospital, and 2 Department of Neurology, Graduate School of Medical Sciences, Kumamoto
University, Japan
No. 15. Preparation of multi-purpose matrix-based material for laboratory service and investigation
Sumiyoshi Naito 1 , Kazumi Akasaska 3 , Shuichi Kino 3 , Yutaka Tomoda 3 , Tadashi Matsubayashi 4 ,
Nobuyuki Kubota 2 , Junichi Ando 5 , Shigemi Hosogaya 6 , Yoshihisa Itoh 1 .
Asahikawa Medical University 1 , Eiken Chemical Co., Ltd. 2 , Asahikawa Medical University
Hospital 3 , Nissui Pharmaceutical Co., Ltd. 4 , SRL, Inc. 5 , Kagawa Prefectural University of Health
Sciences 6 , Japan
No. 16. Temporal changes in estimated glomerular filtration rate (delta eGFR) may be a predictor of
progression to coronary heart disease
So-Young Kim, Tae-Dong Jeong, Woochang Lee, Sail Chun, Won-Ki Min*
Department of Laboratory Medicine, Asan Medical Center and University of Ulsan College of
Medicine, Seoul, Korea
Hitoshi Ikeda (Chairperson)
No. 17. Development of C-reactive protein assay for human saliva
Yumi Narita, Taku Shirakawa
Department of Biophysics, Graduate School of Health Sciences, Kobe University, Japan
No. 18. IODINE STATUS AMONG THE SCHOOL CHILDREN OF HILLY AND PLAIN REGION
OF EASTERN NEPAL
Baral N 1 , Khatiwada S 1 , Shakya PR 1 , Sah GS 2 , Gelal B 1 , Das BKL, Lamsal M 1
1 2
Department of Biochemistry, Department of Pediatrics and Adolescent Medicine, B. P. Koirala
- 37 -

Institute of Health Sciences, Dharan, Nepal
No. 19. Lipid Profile In Menopausal women of Difference Parity and Basal Metabolic index status in
South west, Nigeria.
Adesina Adeyemi Adeleke; Ogunlewe Jayeola; Oyedeji Samuel
Department of Chemical Pathology,OAUTHC,Ife,PMB5538,Osun-state,Nigeria
No. 20. Plasma free Fatty Acid Levels In Normo and Hypertensive Pregnant women before and 72
Hour after delivery in Osun-state Nigeria
Adesina Adeyemi Adeleke; Oyedeji Samuel;Taiwo Funke
Department of Chemical Pathology,OAUTHC,Ife,PMB5538,Osun-State,Nigeria
No. 21. Lipid Peroxidation and Selected Non-Enzymatic Antioxidants in Sickle Cell Disease Patients in
South West, Nigeria
Adesina Adeyemi Adeleke; Oyedeji Samuel; Oladepo Ridwan; Oke Taiwo; Fasakin Kolawole
Department Of Chemical Pathology, OAUTHC, Ife, PMB5538, Osun-State, Nigeria.
Department of Heamatology and Blood Transfusion, OAUTHC, Ife, Osun-State, Nigeria
2. Endocrinology (PE 1-PE 5)
Kuniaki Saito (Chairperson)
No. 22. Two-Step Biochemical Differential Diagnosis of Classic 21-hydroxylase Deficiency and
Cytochrome P450 Oxidoreductase Deficiency in Japanese Infants Using Urinary Steroid
Metabolites by Gas Chromatography - Mass Spectrometry
Yuhei Koyama 1)2) , Keiko Homma 3) , Maki Fukami 4) , Masayuki Miwa 5) , Kazushige Ikeda 5) , Tsutomu
Ogata 4) , Tomonobu Hasegawa 5) , Mitsuru Murata 1)
1) Department of Laboratory Medicine, Keio University School of Medicine, 2) Mitsubishi
Chemical Medience Co., 3) Central Clinical Laboratories, Keio University Hospital, 4) Department
of Endocrinology and Metabolism, National Research Institute for Child Health and Development,
5) Department of Pediatrics, Keio University School of Medicine, Japan
No. 23. Total cholesterol/HDL cholesterol ratio and LDL cholesterol/Apo B ratio as risk factors for
cardiovascular disease in prediabetic of first-degree relatives from patients with type 2 diabetes
mellitus.
Bety Kus Rini Sari, Marzuki Suryaatmadja
Department of Clinical Pathology, Medical Faculty, Universitas Indonesia
No. 24. Adiponectin HMW Level and Homeostasis Model Assessment-Insulin Resistance (HOMA-IR)
- 38 -

in Type 2 Diabetes Mellitus First Degree Relatives
Yulia Sari, Marzuki Suryaatmadja, Budiman Darmowidjojo
Department of Clinical Pathology, Medical Faculty, Universitas Indonesia
No. 25. Age-related Anti-Müllerian hormone Concentrations in Healthy Korean Females
Lee, A 1 ; Jang, TG 2 ; Lim, SY 2 ; Park, JC 2 ; Rhee, JH 2 ;Kim, YJ 1 ; Lee, KY 1
Seoul Medical Science Institute/Seoul Clinical Laboratories, Seoul, Korea 1 , Department of
Obstetrics and Gynecology, School of Medicine, Keimyung University, Daegu, Korea 2
No. 26. Analysis of von Willebrand Factors’ Level in Type 2 Diabetes Mellitus
Mansyur Arif 1 , Ichwan Meinardi 1 , Uleng Bahrun 1 , Harsinen Sanusi 2
1 Department of Clinical Pathology, 2 Department of Internal Medicine, Faculty of Medicine
Hasanuddin University, Makassar, Indonesia
3. Immunology and Blood bank (PI 1 - PI-8)
Seiji Kawano (Chairperson)
No. 27. Combination of Anti-HCV Assay to Improve Performance of Hepatitis C Screening Test in
Prodia National Reference Laboratory, Indonesia
Romimatul Fadhilah, Yenny Surjawan, Indriyanti R. Sukmawati
Prodia Clinical Laboratory, Indonesia
No. 28. Diagnostic value of AFP-L3 and PIVKA-II in hepatocellular carcinoma according to the level
of total-AFP
Eun-Jee Oh 1 , Seung Won Jung 1 , Jong Young Choi 2 , Hee Yeon Kim 2 , Myungshin Kim 1 , Yonggoo
Kim 1 , Dong Goo Kim 3
Department of Laboratory Medicine 1 , Internal Medicine 2 and Surgery 3 , School of Medicine, The
Catholic University of Korea, Korea
No. 29. Development of ABO and Lewis typing by enzyme-linked immunosorbent assay for disaster
Terutaka Sagawa and Mariko Okada
Division of Biomedical Sciences, Institute of Medical Technology, Ehime Prefectural University Of
Health Sciences, Japan
No. 30. Evaluation of Hepatrio multiplex kit for early and simultaneous detection of hepatitis A, B and
C
Chi Hyun Cho, M.D. 1 , Sang Kyung Jo, M.D. 2 , Young Joo Kwon, M.D. 2 , Chae Seung Lim, M.D. 1
and Yun Jung Cho, M.D. 1*
- 39 -

1 Department of Laboratory Medicine, College of Medicine, Korea University, Seoul, Korea
2 Department of Nephrology and Hypertension, College of Medicine, Korea University, Seoul,
Korea
No. 31. Impact of interleukin-29 on interferon alpha secretion of plasmacytoid dendritic cell
Chi Hyun Cho, M.D. 1 , Chae Seung Lim, M.D. and Yun Jung Cho, M.D. 1*
1 Department of Laboratory Medicine, College of Medicine, Korea University, Seoul, Korea
Testuo Kubota (Chairperson)
No. 32. Discrepancy between serologic and genotyping results of ‘Mi a ’ blood antigen and antiserum in
Taiwan
Chien-Feng Sun 1,2 , Tai-Di Chen 2 , Ding-Ping Chen 2
1 Department of Anatomic Pathology, 2 Department of Laboratory Medicine, Linkou Medical Center,
Chang Gung Memorial Hospital, Taoyuan, Taiwan
No. 33. Immune response to MSP2 antigen of plasmodium falciparum in mamuji, west Sulawesi,
Indonesia
Nurhayana Sennang 1 , Dianawaty Amiruddin 2 , Bahrani 2 , Sitti Wahyuni 2 , Syafruddin 2,4 , Irawan
Yusuf 3 , Stephen Rogerson 5
1 2 3
Clinical Pathology Department, Parasitology Department, Physiology Department, Faculty of
Medicine, Hasanuddin University, Makassar, South Sulawesi, Indonesia; 4 Eijkman Institute, Jakarta,
Indonesia; 5 Department of Medicine, University of Melbourne, Post Office Royal Melbourne
Hospital, Melbourne, Victoria, Australia.
No. 34. Inhibition of the NF-kappa B pathway as a candidate strategy for treatment of
cryopyrin-associated periodic syndrome
Satoka Takahashi
Saitama Medical University, Japan
4. Hematology (PH 1-PH 17)
Mitsuhiro Uchiba (Chairperson)
No. 35. Gene Diagnoses of Tumors of Hematopoietic Organs - Diagnosis of Leukemia by both
Southern Method and PCR Method –
Kazuhiro Kabe
Business Department of Medical Technology, Laboratory of Medical Technology, Medic21
Association, Japan
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No. 36. Identification of autoantibodies expressed in acquired aplastic anaemia
Kageaki Kuribayashi 1,2 , Maki Goto 1 , Yusuke Takahashi 1,2 , Takashi Kondoh 1,2 , Maki Tanaka 1,2 ,
Daisuke Kobayashi 1,2 , Naoki Watanabe 1,2,
1 Department of Clinical Laboratory Medicine, Sapporo Medical University School of Medicine,
Japan
2 Division of Laboratory Diagnosis, Sapporo Medical University Hospital, Japan
No. 37. Clinical significance of overexpressed serine-threonine tyrosine kinase 1 in leukemia
Daisuke Kobayashi, Takashi Kondoh, Maki Tanaka, Kageaki Kuribayashi Naoki Watanabe
Department of Clinical Laboratory Medicine, Sapporo Medical University, School of Medicine,
Japan
No. 38. Contribution of uncontrolled fibrinolysis to bleeding diathesis in aortic aneurysm
Mitsuhiro Uchiba, Yuji Yonemura, Yukio Ando
Department of Blood Transfusion and Cell Therapy, Kumamoto University Hospital Kumamoto,
Japan
Naotaka Hamasaki (Chairperson)
No. 39. Aberrant DNA methylation of HOXA4 is associated with resistance to imatinib mesylate in
chronic myeloid leukemia patients
Ravindran Ankathil 1 , Marjanu Hikmah Elias 1 , Azlan H 2 , Sarina Sulong 1 , RoslineHassan 3 , Goh Ai
Sim 4 , S Fadilah Abdul Wahid 5 , Abdul Aziz Baba 2 .
1 2 3
Human Genome Centre, Haemato-Oncology Unit, Department of Internal Medicine, Hematology
Department, School of Medical Sciences, Health Campus, Universiti Sains Malaysia 4 Hospital
Pulau Pinang, Malaysia, 5 Medicine Department & Cell Therapy Centre, UKM Medical Centre,
Malaysia
No. 40. Red Blood Cells absorb and store DARC-affinity chemokines
Hiroyuki Kayaba 1) , Toshihiro Shiratori 1) , Yumiko Yamauti 3) , Norihiro Saito 2) , Junichi Chihara 3) ,
Mihoko Kushibiki 1) , Hiroyuki Akimoto 1) , Shoji Tsutaya 1) , Keiya Kojima 1)
Clinical Laboratory, Hirosaki University Hospital 1) , Yokote Municipal Hospital 2) , Japan
Central Clinical Laboratory, Akita University Hospital 3) , Japan
No. 41. Asian Thrombophilia: Dysfunction of the APC anticoagulation system.
Naotaka Hamasaki
Department of Clinical Chemistry, Faculty of Pharmaceutical Sciences, Nagasaki International
University, Japan
- 41 -

No. 42. Great impact of the brand-new Sysmex XN-Series automated hematology analyzer on
workflow efficiency and utility
Etsuko Hamada, Masato Maekawa
Department of Laboratory Medicine, Hamamatsu University School of Medicine, Hamamatsu,
Japan
Nobuo Masauzi (Chairperson)
No. 43. The comparison of the notations for quantitative evaluation of adhesive molecules’ expression
on CD34 positive cells
Nobuo Masauzi 12 , Junji Tanaka 2 , Masaharu Kasai 3 , Masahiro Ogasawara 3 , Minami Doi 4 , Naoki
Kobayashi 3 , Yoshio Kiyama 3 , Minami Doi 4 , Sayaka Fukui 4 , Nao Fujimoto 4 , Misaki Yamada 4 ,
Keiko Miwa, 1 , Masanobu Kobayashi 5 ,Masahiro Imamura 3 .
1: Department of Medical Laboratory Science, Faculty of Health Sciences, Hokkaido University.
Sapporo, 2: Department of Hematology, Hokkaido University Graduate School of Medicine,
Sapporo, 3: Department of Hematology, Sapporo Hokuyu Hospital. Sapporo, 4: Department of
Health Science, School of Medicine, Hokkaido University, Sapporo, 5: Department of Nursing,
Health Science University of Hokkaido, Ishikari, Japan.
No. 44. Relation between VKORC1 and CYP2C9 polymorphisms and warfarin dose requirement in
Japanese patients
Takeda Tomohiro, Mika Kataoka, Naoko Yamaguchi, Tizuko Kuramoto, Takao Uchiike ,Yasuyuki
Okamoto,
Central Clinical Laboratory, Nara Medical University Hospital, Japan
No. 45. T-cell Acute Lymphoblastic Leukemia with Chromosomal Rearrangements Involving the
Immunoglobulin Heavy Chain Breakpoint at Band 14q32
Joonhong Park 1 , Myungshin Kim 1 , Jihyang Lim 1 , Yonggoo Kim 1 , Kyungja Han 1 , and Seok Lee 2
Department of Laboratory Medicine 1 , Division of Hematology 2 , Department of Internal Medicine,
The Catholic University of Korea, Seoul, Korea
No. 46. Suspect of malaria in Sysmex XT series; A case report of two cases
Iwan Joseph, Mansyur Arif, Hardjoeno
Department of Clinical Pathology, Faculty of Medicine Hasanuddin University, Makassar,
Indonesia
Katsuyasu Saigo (Chairperson)
No. 47. Usefulness of platelet most frequent volume for platelet size evaluation in thrombocytopenic
patients
Setsuki Isono 1) , Megumi Tatsuta 1) , Keiko Nagata 1) , Keiko Numata 1) , Tosiaki Koujitani 1) , Akiharu
- 42 -

Okamura 1) , Masahumi Takata 2) , Mariko Okano 3) , Humiko Hatano 3) , Hiroyasu Uemura 3) , Takeshi
Morisawa 3) , Masahiko Yonetani 3) , Mariko Takenouchi 4) , Katsuyasu Saigo 4)
1) Department of Laboratory Medicine, Kakogawa West City Hospital, Kakogawa, Hyogo. 2)
Department of Internal Medicine, Kakogawa West City Hospital, Kakogawa, Hyogo. 3) Department
of Pediatrics, Kakogawa West City Hospital, Kakogawa, Hyogo. 4) Faculty of Pharmacological
Sciences, Himeji Dokkyo University, Himeji, Hyogo, Japan.
No. 48. Intravascular large B-cell lymphoma in Korea: Clinical and pathological findings
Hyun-Ki Kim 1 , Seongsoo Jang 1 , Chan-Jeoung Park 1 , Hyun-Sook Chi 1 , Jooryung Huh 2 and
Cheolwon Suh 3
Departments of Laboratory Medicine 1 , Pathology 2 and Internal Medicine 3 , University of Ulsan
College of Medicine and Asan Medical Center, Seoul, Korea
No. 49. Evaluation of CellaVision DM96, an automated digital cell morphology identification
system,for the examination of body fluid cytospin slides
Hyun-Sook Chu, Young-Uk Cho, Sang Hyuk Park, Seongsoo Jang, Chan-jeoung Park
Department of Laboratory Medicine, university of Ulsan, College of Medicine and Asan Medical
Center, Seoul, Korea
No. 50. Calculation of Measurement Uncertainty for Complete Blood Cell Counts (CBC)
Atsushi Shirakami, Masako Koguchi, Tsutomu Kakuyama, Kanako Kiso, Takashi Kitagawa,
Sysmex Corporation, Japan
No. 51. Serum Thymidine Kinase 1 as a Potential Marker for Aggressive Behavior in Patients with
B-cell Lymphoma
Heyjin Kim, Jin Kyung Lee, Young Jun Hong, Seok-Il Hong, and Yoon Hwan Chang
Department of Laboratory Medicine, Korea Cancer Center Hospital, Seoul, Korea
5. Clinical Microbiology (PM1-PM14)
Masami Murakami (Chairperson)
No. 52. High seroprevalence of human herpes virus type 8 in patients with lung carcinoma
Cheng-Chuan Su 1,2,3 , Chun-Liang Lai 4 , Ming Nan Lin 5
1
Institute of Medical Sciences, and Departments of Laboratory Medicine and Pathology, School of
Medicine, Tzu Chi University, Hualien, Taiwan; Departments of 2 Clinical Pathology, 3 Anatomic
Pathology, 4 Chest Medicine, and 5 Family Medicine, Buddhist Dalin Tzu Chi General Hospital,
Chiayi, Taiwan
- 43 -

No. 53. HIV-1 RNA viral load in seminal fluid: quantification with NASBA
Lyana Setiawan, Agus Susanto Kosasih, Samsuridjal Djauzi, Haridana Indah Mahdi,
Runingsih
Dharmais Cancer Hospital, Indonesia
No. 54. Epidemiological and microbiological feature of meningococcal infection in Ulaanbaatar city
B.Uyanga, MD 1 , G.Oyungerel, MD 2
1
SOS Medica Hospital, Ulaanbaatar, Mongolia
2
Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
No. 55. The inhibitory effect of serum HLA class I antigens on EBV-specific CD8+CTL in vitro
N.Erdenesuvd 1,3 , B.Gansuvd 1,2 , B.Munkhbat 1,2 , M.Hagihara 2 , T.Hotta 2 , N.Munkhtuvshin 1 ,
1
Central Scientific Research laboratory, Nat’l Institute of Medicine, Mongolia
2
Department of Transplantation Immunology, Tokai University, Japan
3
Dept. Laboratory Medicine, General Hospital, Uvurkhangai, Mongolia
No. 56. Tuberculosis screening by a T cell interferon-γ release assay in students of medical school and
international students in Gunma University
Rumi Watanabe 1) , Takao Kimura 2) , Yutaka Tokue 3) , Takayuki Ogiwara 3) , Makoto Nara 3) , Yoshino
Kobayashi 1) , Toshiya Inoue 1) , Hiroyuki Sumino 1) , Tadashi Morimura 1) , Osamu Araki 1) , Katsuhiko
Tsunekawa 1) , Tomoyuki Aoki 1) , Toshiko Obuchi 3) , Yukie Yomoda 1) , Kihachi Ohshima 2) , Masami
1, 2, 3)
Murakami
1) Clinical Laboratory Center, Gunma University Hospital, Maebashi, Japan. 2) Department of
Clinical Laboratory Medicine, Gunma University Graduate School of Medicine, Maebashi, Japan, 3)
Infection Control and Prevention Center, Gunma University Hospital, Maebashi, Japan.
Hiroaki Onishi (Chairperson)
No. 57. Differences of Vancomycin MICs for MRSA Isolates Based upon the study Susceptibility Test
Method Used
K Watanabe, M Mikami, Y Saya, C Tanaka, F Omata, K Furukawa, K Takeda
St.Luke’s Inte,v,vrnational Hospital, Japan
No. 58. Mycobacterium Kyorinense Infection: Clinical Features and Antimicrobial Susceptibility
Hiroaki Ohnishi 1 , Shota Yonetani 1 , Satsuki Matsushima 1 , Kouki Ohtsuka 1 , Tomonori Kishino 1 ,
Hiroo Wada 2 , Hajime Goto 2 , Takashi Watanabe 1
Departments of 1 Laboratory Medicine and 2 Respiratory Medicine, Kyorin University, School of
Medicine, Japan
No. 59. The first case of Lecythophora hoffmanni peritonitis in Korea
- 44 -

Hunsuk Suh 1 MD, PhD, Jonghee Shin 2 MD,PhD
1.Laboratory medicine, Daegu Catholic University Hospital 2. Laboratory medicine, Jeonnam
University Hospital, Korea
No. 60 Sensitive, one, two-drug resistance, and MDRP Pseudomonas aeruginosa integron carrying rates
Daisuke Tanabe
Bunkyo Gakuin University graduate school, Japan
Satoshi Kimura (Chairperson)
No. 61. Application of MALDI-TOF MS-based strain typing for characterization of epidemiological
relationships among bacterial strains
Megumi Oho 1) , Zenzo Nagasawa 1) , Koji Kusaba 1) , Takanori Higashitani 1) , Shoitiro Ohta 1) , Eisaburo
Sueoka 1) , Hiroshi Miyamoto 2)
Department of Laboratory Medicine, Saga University Hospital 1) , Division of Microbiology,
Department of Pathology and Microbiology, Faculty of Medicine, Saga University 2) , Japan
No. 62. Simultaneous inhibition of NF-κB and caspase-1 by a cell-permeable compound DHMEQ leads
to potent suppression of IL-1β
Sayaka Ito, Misako Iwata, Tetsuo Kubota
Graduate School of Health Care Sciences, Tokyo Medical and Dental University, Japan
No. 63. Comparison of a novel real-time PCR system in the quantification of HIV-1 RNA
Demak Lumban Tobing, Sri Hartini, Theresia Koeshandini, Runingsih
Dharmais Cancer Hospital Clinical Pathology Laboratory, Indonesia
No. 64. Measurement of Procalcitonin (PCT) and C-Reactive Protein (CRP) in Patients with
ESBL-negative Bacteremia
Soohun Yoo 1 , Seri Jeong 1 , Yongjung Park 1 , Nam Su Ku 2 , Dongeun Yong 1 and Hyon-Suk Kim 1*
1 Department of Laboratory Medicine and 2 Internal Medicine, Severance Hospital, Yonsei
University College of Medicine, Seoul, South Korea
No. 65. Significant Increase of Sensitivity on Rapid Influenza Antigen Assays Using Silver
Amplification Immunochromatography
Satoshi KIMURA 1) , Hideyoshi OHTO 2) , Hayato YAMAGUCHI 1) , Seiji FUKUOKA 1) , Emiko
NAKAMA 1) , Akiko TERADA 3) , and Akira UMEDA 2)
1) Department of Laboratory Medicine and Central Clinical Laboratory, Showa University Northern
Yokohama Hospital, Yokohama 224-8503, Japan
2) Department of Pediatrics, Showa University Northern Yokohama Hospital
3) R & D department, FUJIFILM Medical Co., Ltd, Tokyo, 106-0031, Japan
- 45 -

6. Physiological function test (PF1-PF2)
Masato Nishimura (Chairperson)
No. 66. Comparison of four portable apnea recording devices in daytime screening
session for severe sleep apnea patients screening
Chikage Torisawa, Chie Watanabe, Takeshi Tomihara, Chisato Shimazu, Taiji Furukawa
Department of Laboratory Medicine, Teikyo University School of Medicine
No. 67. The relationship between arterial wall elasticity by the phased tracking method
and vascular manifestations in type 2 diabetes mellitus patients
Michiaki Miyamoto 1) , Kazuhiko Kotani 1) , Hideyuki Hasegawa 2, 3) , Hiroshi Kanai 3, 2) , Harumi
Koibuchi 1) , Yasutomo Fujii 1) , Kei Konno 1) , Toshiyuki Yamada 1) and Nobuyuki Taniguchi 1)
1) Department of Clinical Laboratory Medicine, Jichi Medical University, Tochigi, Japan
2) Graduate School of Biomedical Engineering, Tohoku University, Sendai, Japan
3) Graduate School of Engineering, Tohoku University, Sendai, Japan
7. Pathology (PP1-PP7)
Naoki Hosaka (Chairperson)
No. 68. The expressions of O 6 -methylguanine DNA methyltransferase and epidermal growth factor
receptor on ganglioglioma: A clinicopathologic and immunohistochemical study
I-Wei Chang 1,2 , Jui-Wei Lin 3
1
Department of Pathology, E-DA Hospital/I-Shou University
2
Institute of Biotechnology and Chemical Engineering, I-Shou University
3
Department of Pathology , Kaohsiung Chang Gung Memorial Hospital, Taiwan
No. 69. Pathological characteristics of HER2/neu-amplified gastric carcinomas
M. Bamba, H. Sugihara, S. Takemura, K. Fukuda, M. Masuyama, T. Shigematsu and R. Kushima
Depts. Pathology (MB&ST), Surgery (KF&MM) and Gastroenterology (TS), Saiseikai Shiga Hosp.
Dept. Pathology, Shiga Univ. Med. Sci. (HS) and Dept. Pathology, National Cancer Center Hosp.,
Japan
No. 70. Poorly differentiated squamous cell carcinoma of the nipple; A uniqu case for marked
exophytic growth, but little invasion with neuroendocrine differentiation
Naoki Hosaka, Susumu Ikehara, Hakuo Takahashi
- 46 -

Department of Pathology, Kansai Medical University-Kori Hosptal, Department of stem cell
disoders, Department of Clinical Sciences and Laboratory Medicine, Kansai Medical University,
Osaka, Japan
No. 71. Typing of amyloidosis by proteome analysis
Masayoshi Tasaki 1, 2 , Mitsuharu Ueda 3 , Konen Obayashi 3 , Hiroyuki Hata 2 , Hirofumi Jono 3 , Genki
Suenaga 1 , Su Yu 1 , Satoru Shinriki 3 , Taro Yamashita 1 , Yukio Ando 1 1 Department of Neurology,
Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan
2 Department of Immunology and Hematology, Division of Health Sciences, Faculty of Life
Sciences, Kumamoto University, Kumamoto, Japan
3 Department of Diagnostic Medicine, Graduate School of Medical Sciences, Kumamoto University,
Kumamoto, Japan
Tatsuo Sawada (Chairperson)
No. 72. The morphological change after molecular targeting therapy of malignant brain tumor
Tatsuo Sawada 1 , Yoshinori Takekawa 2 , Saori Okamoto 3 , Takashi Maruyama 3 , and Noriyuki Shibata 1
1
Department of Pathology, Tokyo Women’s Medical University, Japan
2
Department of Neurosurgery Tokyo Women’s Medical University, Japan
3
Department of Surgical Pathology Yokosuka Municipal Hospital, Japan
No. 73. Clinicopathological analysis of asbestos-related diseases presenting asbestos bodies in routine
cytology specimens
Makihara K 1) , Kanazawa S 1) , Yoshida T 1) , Sosogi A 1) , Matsuki Y 2) , Shimajiri S 3) , Yamasaki K 4) ,
Hamada T 1)
1) Dept. Surgical Pathology and Asbestos Disease Center, Kyushu Rosai Hospital, Japan
2) Dept. Surgical Pathology, Ootemachi Hospital, Japn
3) Dept. Surgical Pathology, Univ. Occupational and Environmental Health, Japan
4) Dept. Respiratory Medicine, Univ. Occupational and Environmental Health, Japan
No. 74. Neuropathological findings in a case of amyotrophic lateral sclerosis resembling
CIDP
Masaru Yoshioka (1), Hidehiko Konno (1), Toshiaki Takahashi (1), Hiroyasu Tanaka (1), Hiroshi
Onodera (1) , Maki Tateyama (2)
(1) Department of Neurology, National Hospital Organization Nishitaga Hospital, Japan
(2) Department of Neurology, Tohoku University School of Medicine, Japan
8. Molecular Biology (PMB1-PMB12)
- 47 -

Misako Shibakura (Chairperson)
No. 75. Synthesis of cDNA and Detection of Gene Fragment of Protein Disulfide Isomerase Family A
Member 4 (PDIA4) in Bone Marrow Cell
Stefanus Lembar
Atma Jaya Medical Faculty Indonesia
No. 76. Diagnostic strategies lynch syndrome – role of immunohistochemistry and
germline mutation screening -
Mohd Nizam Zahary 1 , Gurjeet Kaur 2 , Muhammad Radzi Abu Hassan 3 , Ahmad Shanwani Mohd
Sidek 4 , Harjinder Singh 5 , Sharifah Emilia Tuan Shariff 6 , Ravindran Ankathil 1 .
1 Human Genome Centre, School of Medical Sciences, Universiti Sains Malaysia Health Campus,
Kubang Kerian, Kelantan, 2 Institute For Research in Molecular Medicine, Universiti Sains Malaysia,
3 Internal Medicine Department, Hospital Alor Setar, Kedah, 4 Surgery Department, Hospital Raja
Perempuan Zainab II, Kota Bharu, Kelantan, 5 Department of Medicine, Hospital Queen Elizabeth,
Kota Kinabalu, Sabah, 6 Department of Pathology, School of Medical Sciences, Universiti Sains
Malaysia Health Campus, Kubang Kerian, Kelantan, Malaysia
No. 77. Corneodesmosin (MHC S) Gene Polymorphism in Mongolian Psoriasis Patients
Ch.Tserendulam 1,3 , B.Munkbat 1,2 , M.Tomizawa 2 , G.Oyungerel 1 , G.Tamiya 2 , Sh.Myagmarjav 4
H.Inoko 2 , N.Munkhtuvshin 1
1 Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
2 Depart. Molecular Life Science, Tokai Uni. School of Medicine, Isehara, Kanagawa, Japan
3 Jargalan Hospital Laboratory, Khuvsgul, Mongolia
No. 78. Study of essential oil for anti-allergy effect in murine asthma model
Tomoe Ueno Iio, Misako Shibakura, Michinori Aoe, Tomoko Hyoda, Mihoko Kohara, Naohisa
Nakagawa, Takako Nakahara, Mikio Kataoka.
Graduate School of Health Sciences, Okayama University., Japan
Yoko Tabe (Chairperson)
No. 79. TGF-β-neutralization enhances cytarabine-induced apoptosis in AML cells in the bone marrow
microenvironment
Yoko Tabe 1,2 , Linhua Jin 1 , Yasuhito Hatanaka 1 , Takashi Miida 1 , Michael Andreeff 2 , Marina
Konopleva 2
1, Department of Clinical Laboratory Medicine, Juntendo University School of Medicine, Tokyo,
Japan
2, Department of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, Texas,
USA
- 48 -

No. 80. Introduction of HCV Quantification as a Diagnostic Tool in Mongolia: Significance and
Lessons Learned
Batmunkh Sayabold 1 , Nemekhbaatar Lkhaasuren 2 , Jazag Amarsanaa 1,5 , Jigjidsuren Chinburen 1,3 ,
Oidov Baatarkhuu 1,2 , Byambaa Suvd 1 , Tseveg Tsogtkhishig 2 , Bira Tsatsralt-Od 1,4 , Batmunkh
Munkhbat 5
1) Mongolian Association for the Study of Liver Diseases, Mongolia; 2) Health Sciences
University of Mongolia, Mongolia; 3) National Cancer Center, Mongolia; 4) National Center of
Communicable Diseases, Mongolia; 5) Institute of Medical Sciences, Mongolia.
No. 81. Development of Thermostabilised MultiplexPCR For Detection of Salmonella Enteritidis and
Vancomycin-Resistant enterococci (VRE)
Tan Ka Liong a , Chan Yean Yean b
a b
Department of pharmacology, and Department of microbiology and parasitology, School of
Medical Sciences, Universiti Sains Malaysia, Malaysia
No. 82. Analysis of Fas exon 6 splicing and gene expression of transcription factors in various human
organs and lymphocyte subsets
Ryo Uozumi 1 , Hitoshi Shibuya 1 , Akio Shigematsu 1 , Kazuhiko Matsuno 1 , Chikara Shimizu 1 , Seiichi
Kobayashi 2 .
1
Department of Clinical Laboratory, Support of Clinical Practice, Hokkaido University Hospital,
2
Department of Medical Laboratory Science, Faculty of Health Sciences, Hokkaido University,
Japan
Kaname Nakatani (Chairperson)
No. 83. The genetic basis of Congenital Hyperinsulinism in Japan
Kumiko Ohkubo 1)2) , Mariko Nagashima 1) , Tomoe Matsuzaki 2) , Makiko Yuki 2) , Hironobu
Kawashima 2) , Akira Matsunaga 1)2)
2) Department of Laboratory Medicine, Faculty of Medicine, Fukuoka University, Japan
3) Department of Clinical Laboratory, Fukuoka University Hospital, Japan
No. 84. A rapid and automated detection system for BRAF mutations in malignant melanoma.
Masaru Ide, Shinichi Koba, Yumi Nagano, Naoko Aragane-Sueoka, Akemi Sato, Takuya Inoue,
Naomi Kobayashi, Noriyuki Misago, Yutaka Narisawa, Shinya Kimura and Eisaburo Sueoka
Faculty of Medicine, Saga University, Japan
No. 85. Detection of Mycobacterium kansasii using amplicons generated by the COBAS TaqMan 48
Analyzer
- 49 -

Arizumi Kikuchi 1 , Yuko Wakayama 1 , Takahiro Sawamura 1 , Satsuki Nakamura 2 , Shu Taga 2 , Yuji
Itoh 2 , Takeshi Seki 2 , Hiroya Sakuma 1,2 , Osami Daimaru 1,2 , Takeo Nakakita 1,2 , Shinichi Itoh 3
1 Daiyukai Second Medical and Science Research Laboratories, 2 Daiyukai General Hospital, 3
Daiyukai First Hospital, Ichinomiya, Aichi, Japan
No. 86. Increased expression of mitochondrial isoenzyme of creatine kinase in hepatocellular
carcinoma cells may be associated with enhanced proliferation and migration
Uranbileg B., Ikeda H., Yatomi Y.
Project researcher, Department of Clinical Laboratory Medicine, Graduate School of Medicine, The
University of Tokyo, Japan
9. Others (PO 1-PO13)
Taiji Furukawa (Chairperson )
No. 87. Application for Diagnostic Tests of Facial Portraits by CCD Camera
Kazuhiro Kabe
Business Department of Medical Technology, Laboratory of Medical Technology, Medic21
Association, Japan
No. 88. Assessment of early manifestation of anthracycline-induced cardiomyopathy in patients with
preserved left ventricular ejection fraction
Reiko Mizuno 1 , Shinichi Fujimoto 2 , Yasuyuki Okamoto 1
1 2
Central Clinical Laboratory, Nara Medical University, Center for Education Development, Nara
Medical University, Japan
No. 89. Influence of masticatory dysfunction on antioxidant capacity in rat model
Maki Tanaka, Kageaki Kuribayashi, Daisuke Kobayashi, Naoki Watanabe
Department of Clinical Laboratory Medicine, Sapporo Medical University School of Medicine,
Japan
No. 90. The role of Rb2/p130 and PP2A interaction in ATRA treatment induced growth arrest of
ovarian carcinoma cells
M.Oyundelger 1,3 , P.Enkhtsetseg 1,2 , Soprano DR 2 , Soprano KJ 2 , N.Munkhtuvshin 1
1
Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
2
Department of Microbiology and Immunology, Temple University School of Medicine,
Philadelphia,
3 CEO Laboratory, Monolab Co., Ulaanbaatar, Mongolia
- 50 -

No. 91. Genome-wide association analysis with selective genotyping identifies candidate loci for adult
height at 8q21.13 and 15q22.33-q23 in Mongolians
M.Enkhdelger 1,3 , T.Kimura 2 , T.Kobayashi 2 , B.Munkhbat 1,2 , G. Oyungerel 1 , H. Hayashi 2 A.Oka 2 , I.
Inoue 2 , H.Inoko 2 , N.Munkhtuvshin 1
1 Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
2 Department of Molecular Life Science, Tokai University School of Medicine, Isehara, Kanagawa,
Japan
3 CEO Laboratory, Monolab Co., Ulaanbaatar Mongolia
Hayato Miyachi (Chairperson)
No. 92. Molecular analysis of HLA polymorphism in ethnic minority of Khoton-Mongolians
P.Nyamragchaa 1,3 , B.Munkhbat 1,2 , T.Sato 2 , M.Hagihara 2 , K.Sato 2 , A.Kimura 2 , K.Tsuji 2 ,
N.Munkhtuvshin 1
1 Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
2 Department of Transplantation Immunology, Tokai University, Japan
3 Dept. Laboratory Medicine, General Hospital, Dundgobi, Mongolia
No. 93. TNF-β gene-polymorphism in population and clinical studies
B.Ankhchimeg 1,3 , B.Munkhbat 1,2 , B.Gansuvd 1,2 , G.Oyungerel 1 , T.Shimura 2 N.Munkhtuvshin 1
1 Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
2 Department of Transplantation Immunology, Tokai University, Japan
3 Dept. Laboratory Medicine, General Hospital, Arkhangai, Mongolia
No. 94. The genealogical study of Charcot-Marie-Tooth disease prevalence among some Families in
Bayan-Uul soum of Gobi-Altai province
Batchimeg B 1 , Bilegtsaikhan Ts 1 , Oyungerel G 1 , Tselmen D 1 , Erdenechimeg Ya 2 , Oyuntsetseg М 3 ,
Baasanjav D 2 , Munkhtuvshin N 1 , Munkhbat B 1
1Central Scientific Research Laboratory, Institute of Medical Sciences, Mongolia
2Department of Neurology, Institute of Medical Sciences, Mongolia
3Department of Neurology, Central Hospital of Gobi-Altai Province
No. 95. Number and morphology of mesothelial cells in peritoneal dialysis effluent as a potential
predictive marker for advanced peritoneal dysfunction
Mayumi Idei 1 , Yoko Tabe 1 , Kazunori Miyake 1 , Chieko Hamada 2 , Hiroyuki Takemura 3 , Hiroaki Io 2 ,
Kiyoshi Ishii 3 , Takashi Horii 3 , Yasuhiko Tomino 2 , Akimichi Ohsaka 3 , Takashi Miida 1
1, Department of Clinical Laboratory Medicine, Juntendo University School of Medicine, Tokyo,
Japan
2, Division of Nephrology, Department of Internal Medicine, Juntendo University School of
Medicine , Tokyo, Japan
- 51 -

3, Division of Clinical Laboratory, Juntendo University Hospital, Tokyo, Japan
Masahiro Koshiba (Chairperson)
No. 96. Regulatory and pro-inflammatory properties of CD4+ T-cell subsets defined by CD45RA,
CCR7, CD27, and CD28 in patients with rheumatoid arthritis
Fumichika Matsuki*†, Jun Saegusa*†, Yoshiaki Miyamoto†, Kenta Misaki†, Shunichi Kumagai*§,
and Akio Morinobu†
Department of Evidence-based Laboratory Medicine, Kobe University Graduate School of
Medicine, Kobe, Japan; †Department of Rheumatology and Clinical Immunology, Kobe University
Graduate School of Medicine, Kobe, Japan; §The Center for Rheumatic Diseases, Shinko Hospital,
Kobe, Japan
No. 97. 5-Ethynyl-2′-deoxyuridine (EdU)-induced replication banding patterns in human chromosomes
Osamu Hoshi
Anatomy and Physiological Science, Graduate School of Health Care Science, Tokyo Medical and
Dental University, Japan
No. 98. Chaotic Nature of Myocardial Ultrasonic Radio Frequency Signals Reflect Cardiac Fibrosis and
Hypertrophy in Patients with Hypertension.
Mitsuru Masaki, *# ; Akiko Goda, * ; Masataka Sugahara, * ; Miho Fukui, *# ; Minoru Murakami, # ; Miho
Kuroda, # ; Ayumi Igaki, # ; Seiji Fujii, # ; Tadanao Wada, # ; Kohji Inuzumi, # , Masahiro Koshiba, # ; and
Tohru Masuyama, *
Cardiovascular Division, Department of Internal Medicine, Hyogo College of Medicine *
Division of Clinical Laboratory Medicine, Hyogo College of Medicine #
No. 99. Drug susceptibility test for Giardia intestinalis using WST-1 reagent
Takahiro Matsumura *,** , Tomoji Yuno * , Masaharu Tokoro **
*
Department of Clinical Laboratory, Kanazawa Red Cross Hospital
**
Department of Parasitology, Graduate School of Medical Science, Kanazawa University, Japan
- 52 -

Abstracts
- 53 -

No. 1 (PC 1)
Tacrolimus in Adult Renal Transplant Recipients
Pradeep Naik, Mallikarjuna Madhavarapu
Department of Clinical Biochemistry, Global Hospitals, Lakdikapool, Hyderabad-500004, India
Background
Success of an immunosuppressive drug therapy depends on extent of exposure to the drugs (the blood
levels and duration) which is measured as area under the curve (AUC). Tacrolimus also shows
considerable variability in its pharmacokinetics, with poor correlation between Tacrolimus trough level
and systemic exposure, as measured by the area under the curve of concentration time. Monitoring
trough levels helps not only in reducing nephrotoxiicity but also the chances of acute rejection; even
though there is no international consensus, the trough concentration is used to determine dosing and the
AUC for calculating the exposure of the patient to the drug. The major objective of this study was to
find the best sampling time for an abbreviated AUC0-6 (area under the concentration time curve) to
predict the total body exposure to Tacrolimus in adult renal transplantation recipients.
Methods
The study involved retrospective analysis of 14 renal transplant patients (2 female+12 male) who were
on triple immuno suppressive therapy, methyl prednisolone, mycophenolate mofetil and Tacrolimus.
Blood samples were collected before administering Tacrolimus (0 h) to determine trough concentration
and at fixed time points of 2h, 4h and 6h after administration of oral Tacrolimus and analyzed
induplicate by micro particle enzyme immunoassay. AUC0-6 was determined using the linear trapezoidal
rule. Association between the blood concentration and AUC6 were evaluated by Pearson correlation
coefficient. All statistical analyses were performed using SPSS software program.
Results
The trough levels were fairly consistent at 7.9-18 ng . h/mL in all the patients included in this study and
this did not show variation with age or sex. The AUC0-6 was higher (202-290 ng/mL at 3-8 mg Bis-Daily
(BD) dosage) in patients who received kidney from cadaver compared to recipients from live donors
(60.5-171 ng/mL at 3-8 mg BD dosage) but whether the clinical significance of this is not known.
Highest AUC0-6 was 246 ng/mL observed at 4.5 mg BD dosage. Dosages higher than 2 mg BD did not
result in noticeable increase in AUC0-6. Peak blood levels of Tacrolimus were obtained 4 h after
administration
Conclusion
Trough level determination and C2, C4 two-point limited sampling strategy may be useful to plan the
dosing strategy and estimate exposure of renal transplant patients to Tacrolimus.
- 54 -

No. 2 (PC2)
Soluble FcγRIIIa Mφ levels in plasma correlate with maximum plaque diameter in
patients examined with carotid arterial echography.
Midori MASUDA 1 , Keiko KOUNO 2 , Noriko NISHIMURA 1 , Hiroya MASAKI 1 , Toyohiko
YOKOI 1 , Masamichi YOSHIKA 1 , Yutaka KOMIYAMA 1 , Toshiji IWASAKA 2 , and Hakuo
TAKAHASHI 1
Departments of 1 Clinical Sciences and Laboratory Medicine, and 2 Medicine II, Kansai Medical
University, Osaka, Japan
Macrophages play a major role in the development of vascular lesions in atherogenesis. The cells
express FcγRIIIa (CD16) identical to that in NK cells, but with a cell type-specific glycosylation, and
these soluble forms (sFcγRIIIa) are present in plasma. We measured sFcγRIIIa Mφ derived from
macrophages in plasma with anti-FcγRIIIa Mφ monoclonal antibodies and found that the levels of
sFcγRIIIa Mφ were related to the severity of coronary atherosclerosis in patients with coronary artery
diseases (CAD).In volunteers, the sFcγRIIIa Mφ levels measured at the annual medical checkup were
related to the number of risk factors for atherosclerosis and were correlated with carotid maximum
intima-media thickness (IMT). In the present study, we measured sFcγRIIIa Mφ in plasma from patients
examined with carotid artery echography. The levels of sFcγRIIIa Mφ , but not the sFcγRIIIa, were
significantly increased in patients examined with carotid arterial echography, compared with
age-matched healthy controls. Patients with echogenic/echolucent plaques or mixed with
echogenic/echolucent and calcified plaques had the higher sFcγRIIIa Mφ levels, but patients with only
calcified plaques had levels similar to patients with no plaque or intact carotid artery. The levels of
sFcγRIIIa Mφ were correlated with maximum plaque diameters, especially in patients with
echogenic/echolucent plaques. Because unstable plaques contain a lot of lipid and macrophages inside,
the sFcγRIIIa Mφ may serve as a novel risk biomarker for unstable angina or myocardial infarction.
- 55 -

No. 3 (PC 3)
The Role of S100B Protein and GFAP in Predicting Discharged NIHSS of Anterior
Circulation Ischemic Stroke
Yenny Surjawan, Suryani As’ad, Teguh A.S Ranakusuma, Andi Wijaya
Medical Faculty of Hasanuddin University, INDONESIA
Background: Patient with larger ischemic lesion will suffer more severe neurological deficit. Lesion
size measurement by MRI is still limited. Another approach pursued through examination of markers
released by damaged brain cell. S100B protein and GFAP, markers of damaged brain cell, had been
reported to have an association with stroke. This study aimed to know whether both markers could
estimate the discharged NIHSS of anterior circulation ischemic stroke.
Methods: This observational prospective study was performed on 74 anterior circulation ischemic
stroke patients. S100B protein and GFAP were measured by ELISA.
Results: A significant difference was found in S100B protein concentration between mild and not mild
discharged NIHSS (p < 0.05). S100B protein concentration showed a significant correlation with
discharged NIHSS (r = 0.488; p = 0.000). An equation was yield to predict the risk and probability to get
a not mild discharged NIHSS by S100B protein (OR = 1.009; 95 % CI 1.0003 - 1.0188; p = 0.044).
S100B protein at 78.3215 ng/l produced a 66 % sensitivity and 70 % specificity, with positive and
negative predictive value of 76 % and 58 %, respectively (AUC = 73.9 %, p = 0.001, 95 % CI: 62.7 % –
85.2 %). There was a significant difference in the proportion of subjects with low and high GFAP in
either not severe or severe group of NIHSS at discharge (p = 0.008).
Conclusion: S100B protein measured at 48 to 72 hours after onset could predict the risk and probability
to get a not mild discharged NIHSS in anterior circulation ischemic stroke patients. No subject with low
GFAP level showed a severe NIHSS at discharge.
- 56 -

No. 4 (PC 4)
Detection of hereditary abnormality of phenylalanine metabolism using
microbiologic method
E.Shuree 1 , G.Oyungerel 2 , N. Munkhtuvshin 2
1
Oyushur Laboratory, Ulaanbaatar, Mongolia
2
Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
Objectives: Mass screening of hereditary abnormality of phenylalanine metabolism in newborn as
essential test procedures in many countries request evaluate feasibility of low cost microbiological test
in screening of children in the country with less developed infrastructure.
Materials and Methods: Blood samples were collected from 620 pupils studied at specialized school
№29 of Ulaanbaatar with clinical diagnosis of mental deficiency, different forms. Genetic defect of
phenylalanine metabolism was investigated by Guthrie test, based on competition between the amino
acid Phenylalanine and inhibitor for phenylalanine ß-2-Thyenilalanine, kindly provided by WHO
country representative from supplier Difco laboratories inc, 920 Henry Street, Detroit, MI 48201-2532.
Results: Phenylketonuria (PKU) is a recessive hereditary disease, with estimated frequency of
1:10.000. Patients have deficient hepatic enzyme Phenylalanine-hydroxylase, which catalyzes the
conversion of Phenylalanine to Tyrosine and consequently an increase of Phenylalanine in blood and an
increase of Phenylpiruvic Acid in urine (PKU), which causes a mental deficiency in new-born infants. A
possibility to avoid mental damages is feeding new-born babies with a Phenylalanine free diet from the
first weeks of life. Mass screening of PKU is not introduced in the country yet. Therefore cost efficient
screening test to diagnose PKU is very useful since it gives a chance to prevent any mental damage. Of
the 620 investigated children (285-girls, 345- boys within the age of 8-18) positive results were obtained
in 10 mentally retarded children (1.63%).
Conclusion: 1. Microbiological method of Guthrie is semi-quantitative determination of phenylalanine
in the blood and characterized by high sensibility and reproducibility and feasible test for screening of
children in countries with limited resources. 2. The prevalence of positive results was almost 2 and half
times more in comparison to the results of foreign investigators, where used biochemical urine test and
where PKU is revealed in 7 of 1125 mentally retarded patients (0.6%).
- 57 -

No. 5 (PC 5)
External quality assessment schemes for Clinical Chemistry in Mongolia
G.Oyungerel 1 , Ya Amarjargal 2 , N.Munkhtuvshin 1
1
Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
2
Dept. Medical Care Policy Implementation and Coordination, MOH, Mongolia
Objectives: The main objective of NEQAS in Mongolia is to provide an educational support towards
improvement of clinical chemistry laboratory diagnostic services and participation is voluntary and
confidential, though State Inspectorate makes participation obligatory for both governmental and private
laboratories.
Materials and methods: Lyophilized commercial QC materials for 20 parameters and structured report
format distributed to the participants monthly and the Scoring System (Bullock & Wilde, 1985) used for
calculation. The system depends on number of related indices such as Variance index score (VIS), the
difference, between the result returned and the designated value, expressed as a percentage of the
designated value, divided by the chosen coefficient of Variation (CCV) for the analyte, expressed as
percentage, and OMRVIS, the mean of the 40 most recent variance index scores irrespective of analyte
for each laboratory.
Results: CSRL, NIMM, approved by the Health Minister, Mongolia as principal organizer of the
NEQAS and reflects the WHO recommendations on EQAS design and purpose and the scheme provide
assessments of the state of the art (general performance) of both individual laboratories and countrywide
services and the effects of individual analytical procedures (method principles, reagents and
instruments). Doubled decrease (improvement) of countrywide OMRVIS achieved in 2011since 2000
(from 320 to 160) and dramatic decrease of MRVIS observed in individual best laboratories during this
period of time (from 220 to 55).
Conclusions: System of Clinical Chemistry NEQASs of Mongolia has been introduced since over more
than 20 years and its usefulness is supported by the remarkable evidence of slow but continuing
improvement of performance throughout the country, having importance in facilitating the provision of
reliable diagnostic services.
- 58 -

No. 6 (PC 6)
Analysis of cholesteryl ester hydroperoxides in oxidized lipoprotein by Use of an
Orbitrap LC-MS
Shu-Ping Hui 1 , Toshihiro Sakurai 1 , Futaba Ohkawa 1 , Hiroaki Furumaki 1 , Shigeki Jin 1 , Hirotoshi
Fuda 1 , Seiji Takeda 1 , Takao Kurosawa 2 , Hitoshi Chiba 1
1 2
Faculty of Health Sciences, Hokkaido University, Kita-12, Nishi-5, Sapporo 060-0812, Japan Faculty
of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido
061-0293, Japan
Oxidation of cholesteryl esters in lipoproteins by reactive oxygen species yields cholesteryl ester
hydroperoxides (CEOOH). In this study, we developed a novel method for identification and
characterization of CEOOH molecules in human lipoproteins by use of reversed-phase liquid
chromatography with an hybrid linear ion trap–Orbitrap mass spectrometer (LC–LTQ Orbitrap).
Electrospray ionization tandem mass spectrometric analysis was performed in both positive-ion and
negative-ion modes. Identification of CEOOH molecules was completed by use of high-massaccuracy
(MA) mass spectrometric data obtained by using the spectrometer in Fourier-transform (FT) mode.
Native low-density lipoproteins (nLDL) and native high-density lipoproteins (nHDL) from a healthy
donor were oxidized by CuSO4, furnishing oxidized LDL (oxLDL) and oxidized HDL (oxHDL). No
CEOOH molecules were detected in the nLDL and the nHDL, whereas six CEOOH molecules were
detected in the oxLDL and the oxHDL. In positive-ion mode, CEOOH was detected as [M + NH4] + and
[M + Na] + ions. In negative-ion mode, CEOOH was detected as [M +CH3COO] − ions.
CEOOH were more easily ionized in positive-ion mode than in negative-ion mode. The LC–LTQ
Orbitrap method was applied to human plasma and six species of CEOOH were detected. The limit of
detection was 0.1 pmol (S/N=5:1) for synthesized CEOOH.
- 59 -

No. 7 (PC 7)
An approach to chemical education at medical technologist training institutions in
Japan
Hidetusgu Kohzaki
Department of Cell Biology, Institute for Virus Research, Kyoto University, Japan
I teach chemistry at a medical technology school (1, 2, 3, 4). Chemistry is not necessarily the favorite
subject of Japanese students receiving paramedical education (1). However, many biochemical,
microbiological, and advanced genetic tests (3, 4) are performed in the medical setting, and solutions
and media also have to be prepared. Nurses and paramedics including medical technologists rarely
prepare solutions because of automation in the medical setting and a lack of time. However, they might
feel uneasy about certain aspects of medical practice, such as the use of infusions and injections, if they
do not know how to prepare and label solutions. In addition, the annual national examinations include
questions on concentration calculations. In this article, I introduce our approach to providing chemical
education for medical technologists.
1. Kohzaki, H. A proposal of chemistry education for medical technologist/paramedics in Japan.
Chemical Education Journal 14, 3, 2011. (http://chem.sci.utsunomiya-u.ac.jp/v14n1/kohzaki/)
kohzaki. html)
2. Kohzaki H. A proposal regarding English education at schools to train paramedics/medical
technologists in Japan. Journal of Medical English Education. J. Med. Eng. Edu. 11(1), 7-14, 2012.
3. Kohzaki H. Detection of chromosomal abnormality and mutations by Chromatin
immunoprecipitation (ChIP) assay. J. Chromosome and Gene Analysis 28:122-126, 2010.
4. Kohzaki H. A proposal of chromosome and gene analysis education in training institutions for
medical technologists and an idea of National examinations for them. J. Chromosome and Gene
Analysis. 30:68-74, 2012.
- 60 -

No. 8 (PC 8)
Development of the enzymatic method of serum ethanolamine and examination of
its clinical utility
E. Ohta (1) , E.Hokazono (1) , M.Ono (2) , T.Hotta (2) , S.Osawa (3) , Y.Kayamori (1)
1
Division of Medical Technology, Department of Health Sciences, Graduate school of
Medicine, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka City, Japan
2
Department of Clinical Chemistry and Laboratory Medicine, Kyushu University
Hospital, 3-1-1 Maidashi, Higashi-ku, Fukuoka City, Japan 3 Division of Clinical
Laboratory Science, Department of Medical Risk And Crisis Management, Chiba
Institute of Science, 15-8 Shiomi-cho, Choshi, Chiba, Japan
Ethanolamine (EA) is mainly hydrolyzed from phosphatidyl ethanolamine (PE) by phospholipase D
(PLD) in vivo. A study reported that urine EA increased in newborns as Zellweger syndrome, a
congenital metabolic disease, but its concentration in blood and clinical significance in adults were not
clarified. Recently, a metabolomics study using mass spectrometry reported that EA in saliva of
pancreatic cancer patients increased significantly. Therefore, our purposes are to develop a rapid and
simple enzymatic method with an amine oxidase involving copper from Arthrobacter sp. (AAO) (EC
1.4.3.6) and to examine the clinical meaning of EA in serum. In our measurement method, reagent 1
(R-1) contained 0.1 mol/L HEPES buffer (pH 8.2 at 25ºC), 1.6 mmol/L TOOS, 5.0 kU/L POD, 1.12
mmol/L N-ethylmaleimide, 16.8 kU/L L-ascorbate oxidase (ASOD) and 7.47 mmol/L NaN3. Reagent 2
(R-2) contained 0.1 mol/L HEPES buffer (pH 8.2 at 25ºC), 20.0 kU/L AAO and 0.4 mmol/L
4-aminoantipyrine. The assay used a Hitachi 7170 type analyzer. A 10 µL specimen was mixed with 180
µL R-1; after incubation for 5 min at 37 ºC, 90 µL R-2 was added; after another 5 min, the mixture was
measured using a 2-point end assay performed at 37 ºC, with wavelengths of 700/546 (sub/main
wavelength). The within-run CVs of the present method with three kinds of EA solutions ranged
0.43-7.13% (n=10). The standard curve showed linearity from 0 to 800 µmol/L. Analytical recovery was
98.4%. The reference interval of EA in normal was 8.17 ± 4.83 µmol/L (n=19). EA in hepatocellular
carcinoma patient sera was significantly higher than normal. In conclusion, our method for serum EA is
suitable for routine clinical use in the laboratory for determination of accuracy and simplicity. We
consider EA in serum may be a new biomarker of hepatocellular carcinoma.
- 61 -

No. 9 (PC 9)
Establishment of enzyme-linked immunosorbent assay for urinary Tamm-Horsfall
protein
Aki Nakayama 1 , Ryo Kubota 2 , Minoru Sakatsume 3 , Shiro Iijima 1,4 , Kiyoko Shiba 4
1. Faculty of Health Science Technology, Bunkyo Gakuin University, Japan
2. Department of Health Sciences, Saitama Prefectural University, Japan
3. Division of Clinical Nephrology and Rheumatology, Graduate School of Medical and
Dental Sciences, Niigata University, Japan
4. Graduate School of Health Care Science, Bunkyo Gakuin University, Japan
Background: The Tamm-Horsfall protein (THP) is a 80–90 kD glycoprotein synthesized in the thick
ascending limbs of the Henle’s loop in the kidney. Upregulation- or downregulation of this protein’s
expression is observed in patients with several renal diseases. THP aggregates with itself or other
urinary components. To dissociate this, the assay procedures require urine samples be highly diluted
(>100-fold). Therefore, we aimed to develop a highly sensitive enzyme-linked immunosorbent assay
(ELISA) for determining urinary THP levels, and investigated the usefulness of measuring urinary THP
levels in diagnosing renal dysfunction.
Methods: THP was purified from pooled urine samples of healthy subjects by performing diatomaceous
earth filtration. This THP was used as an antigen in mice to raise monoclonal antibodies by using classic
hybridoma techniques, and used for the capturing of the antibody in the ELISA. Polyclonal antibody
was also raised in rabbits, and used for the detection antibody. We analyzed 25 samples from untreated
patients with different kinds of glomerulonephritis and age-matched healthy subjects. The urine samples
were diluted to ~1:10000 with distilled water and with an alkaline diluent to inhibit THP polymerization
under acidic urine conditions.
Results: The range of the ELISA calibration curve was 0–40 μg/L, and the lower detection limit was
0.313 μg/L. The dilution curves of urine samples showed good linearity. The within-run CV and the
between-day CV were less than 8%. The assay recovery was 81–123%. The urinary THP concentrations
of the patients with glomerulonephritis (10.8 ± 2.04 mg/L) were significantly lower than those of the
healthy subjects (49.2 ± 5.48 mg/L).
Conclusion: The newly developed ELISA showed high sensitivity for detecting THP in highly diluted
urine samples and showed that urinary THP levels could aid in diagnosing renal dysfunction.
- 62 -

No. 10 (PC 10)
Direct analysis of chemical substances in biofluids without sample pretreatment
Ritsuko Wakayama 1 , Yasushi Iwasaki 1 , Motoaki Kamachi 1 , Natsumi Sawa 2 , Katsuko Hara 2 ,
Yutaka Komiyama 2
1 Showa Denko K.K, Japan, 2 Department of Clinical Sciences and Laboratory Medicine, Kansai Medical
University Takii hospital, Japan
Sample pretreatment is often a time consuming and laborsome step which can be a restricting factor for
the rapid analysis. This is especially true for the analysis of drugs in biofluids containing many
interfering matrices. Therefore, we developed a direct analysis method for drugs in biofluids, using a
polymer-based reversed-phase HPLC column which has a unique feature of retaining hydrophilic target
compounds while excluding large biological matrices prior to the elution of targets. Following
conditions were used. Column; Shodex ODP2 HP-4D (4.6 mm ID x 150 mm L) and ODP2 HP-4E (4.6
mm ID x 250 mm L) from Showa Denko K.K. Eluent; 0.1% TFA/CH3CN = 93/7. Flow rate; between
0.3 and 0.6 mL/min. Injection volume; from 5 to 10 uL. Detector; UV with wavelength between 215 and
272 nm. Column temperature; 40°C. Standards were prepared in the eluent. Optimal conditions for each
target and biofluids varied slightly. For the identification purpose, each peak were fractionated, and then
injected directly to a MS/MS. The components of typical cold medicines were detected without the
influence of biological matrices. Blood serum, urine, and gastric juice samples were collected from
suspected drug-poisoned patients transported to the emergency and critical care medicine. Several
components in cold medicine were detected in one patient’s serum: 44.1 ug/mL acetaminophen, 11.0
ug/mL dihydrocodein phosphate, and 22.6 ug/mL aspirin. A small amount of salicylamide and salicylic
acid was also found. Theophylline and risperidone were detectable in other patients’ serum samples. In
one case of accidental insecticide poisoning, imidacloprid was detected in the patient’s gastric juice.
The presented method using the ODP2 HP column provides a simple and rapid analysis of drugs in
biofluids without the necessity of complex sample pretreatment. This method is applicable for the
diagnosis and treatment of patients who are poisoned or overdosed counter medicine.
- 63 -

No. 11 (PC 11)
Indoxyl Sulfate is an Independent Risk Factor for Coronary Heart Disease in
Patients with Chronic Kidney Disease
Ippei Watanabe 1 , Shunji Namba 2 , Yuko Okuda 2 , Akiko Morimoto 2 , Tetsumi Kojima 2 ,
Masanari Sano 2 , Toshisuke Morita 1
1. Deartment of Laboratory Medicine Graduate School of Medicine Toho University, Japan
2. Clinical Laboratory Toho University Omori Medical Center, Japan
Background: Chronic kidney disease (CKD) has been recognized as an independent risk factor for
coronary heart disease (CHD). Indoxyl sulfate (IS), one of uremic toxins, may play an important role
in cardio-renal syndrome and our previous in vitro study showed that IS mediates vascular toxicity
through enhanced oxidative stress in human endothelial cells. However, it is still unclear how
accumulated IS in patients with renal dysfunction affects clinical outcome in those patients. In the
present study, we examined the relation between CHD and serum IS levels in patients with CKD.
Methods: In November 2010, patients in different CKD stages were divided into two groups (CHD
group n=50 and non-CHD group n=62). CHD was diagnosed by coronary angiography, nuclear
cardiac scintigraphy or multi-slice CT. The background and clinical characteristics of two groups were
compared using receiver-operating characteristic analysis and multivariate analysis. The cut-off value of
serum IS level was calculated to achieve its risk index on CHD.
Results: Serum IS levels were significantly higher in the CHD group than in the non-CHD group
(P

No. 12 (PC 12)
Reference value for serum folic acid level in Japanese adults
Naoko Yamaguchi, Tomohiro Takeda, Chizuko Kuramoto, Takao Uchiike, Masaharu Yamazaki,
and Yasuyuki Okamoto
Central Clinical Laboratories, Nara Medical University, Japan
Since folic acid deficiency in early pregnancy may cause neural tube closure defect in embryos,
Japanese Ministry of Health, Labour and Welfare recommends a daily intake of 0.4 mg folic acid for
women who can become pregnant. It has been believed that the food folate is generally sufficient in
Japanese adults, however, it has been suggested that recent changes in domestic eating habits might
have reduced the intake. Thus, we measured folic acid in sera of Japanese healthy adults using a
chemiluminescent enzyme immunoassay (CLEIA), and here report the reference value obtained as a
result.
Materials and Methods
Three hundreds and fifty-nine healthy volunteers in our hospital were enrolled in the study. Serum
folic acid, vitamin B12 and ferritin were measured on an Access2 analyzer (Beckman-Coulter Inc.).
Results
The reference value between the 2.5th and 97.5th percentiles in the frequency distribution of serum
folate levels was determined as 2.41-11.93 ng/mL. Median values for serum folate levels were: 4.16
ng/mL in males, 4.75 ng/mL in females, and 4.57 ng/mL in all subjects. Serum folate levels tended to be
higher in females and lower in younger subjects.
Discussion
Our reference value for serum folate levels is similar to that reported previously in the UK using the
same analyzer. The finding that serum folate levels are higher in female subjects is also consistent with
the previous reports. The reason why serum folate levels were lower in our younger subjects is
unknown, but might be explained by differences in eating habits between generations. We are preparing
to report a further study on serum folate levels in pregnant women and anemic patients.
- 65 -

No. 13 (PC 13)
Effects of alpha lipoic acid as an antioxidant in preventing renal mitochondrial
oxidative damage in diabetic rats
Noraihan Mat Harun, Rahajoe Imam Santosa
Department of Basic Medical Sciences, Kulliyyah of Medicine, International Islamic University,
Malaysia, Kuantan-Malaysia
Diabetes mellitus is a metabolic disorder characterized by chronic hyperglycemia due to inadequate
secretion of insulin or insufficient action of the endogenous insulin. Oxidative stress has an important
role in the pathogenesis of diabetic complications and mitochondria represent a major source of the
reactive oxidants that lead to cellular oxidative damage. This study is design to observe the effect of
alpha lipoic acid (ALA) as mitochondrial antioxidant in preventing renal mitochondrial oxidative
damage in diabetic induced rats. In this study, the rats were divided into three groups. Normal Control
(NC) group was served as a control; Diabetic Control (DC) group was induced for diabetes without
given any supplementation, while Diabetic-Alpha Lipoic Acid (D-ALA) group was
streptozotocin-induced diabetic rats, supplemented with ALA daily for six weeks. The ability of ALA in
preventing renal mitochondrial oxidative damage was evaluated by comparing the renal function in
terms of serum urea, serum creatinine and creatinine clearance in both supplemented and
non-supplemented rats, as well as mitochondrial H2O2 concentration and GSH/GSSG ratio as parameters
for the oxidative stress. The results showed that ALA capable to ameliorate the defective antioxidant
defense system leads to modulate the oxidative stress in terms of lower H2O2 concentration and higher
GSH/GSSG ratio in D-ALA group compared with DC group. However the significant reduction of
oxidative stress did not correlate well with the renal function parameters, in which only showed
significant improvement of serum urea, but not serum creatinine or creatinine clearance. In conclusion,
ALA treatment capable to ameliorate the defective antioxidant defense system and modulate the
oxidative stress, but partially preventing the progression of renal dysfunction in diabetic rats.
- 66 -

No. 14 (PC 14)
Detection of autoantibodies against ATTR in patients with FAP ATTR V30M
Konen Obayashi 1 , Masayoshi Tasaki 2 , Satoru Shinriki 1 , Genki Suenaga 2 and Yukio Ando 2
1
Diagnostic Unit for Amyloidosis, Department of Laboratory Medicine, Kumamoto University Hospital,
and 2 Department of Neurology, Graduate School of Medical Sciences, Kumamoto University, Japan
Background: It has been well documented that conformational changes via instability of tetrameric form
TTR occurs during the process of amyloid formation in FAP. Some cryptic epitopes expose on
amyloidogenic TTR (ATTR) molecule surface during the process in sera of FAP patients. There is a
possibility that de novo antibodies responded to cryptic epitopes of ATTR may be synthesized in serum
of FAP patients. Objective: To examine whether a relationship exists between the incidence of
autoantibodies against ATTR and the phenotype of FAP ATTR V30M. Methods: Serum samples were
collected from 25 Japanese (28-32 years old) and 8 Swedish (67-81 years old) FAP ATTR V30M
patients, 4 Japanese gene carriers of ATTR V30M, and 24 Japanese healthy controls which had no
genetic mutations of TTR. To detect the presence of autoantibodies against ATTR in these serum
samples, enzyme-linked immunosorbent assay (ELISA) using recombinant ATTR V30M protein was
immobilized on a polystyrene microtiter plates. Correlation between data of the ELISA and age of the
subjects or duration of FAP was investigated. Results: Three of 25 Japanese and 5 of 8 Swedish FAP
ATTR V30M patients were found to have autoantibodies against ATTR, and these 8 patients were all
late onset cases. Moreover, significant positive correlation between the presence of the antibody and age
was observed in all the patients examined (r=0.77, p < 0.05). We are now trying to detect
conformational epitope binding to the autoantibodies against ATTR V30M. Our preliminary data
suggested the possibility that there were some epitopes at positions 24-35 or 105-115 of ATTRV30M.
Conclusions: Age-dependent increase in the incidence of autoantibodies was observed in patients with
FAP ATTR V30M. This phenomenon may influence the clinicopathological differences between both
early (

No. 15 (PC 15)
Preparation of multi-purpose matrix-based material for laboratory service and
investigation
Sumiyoshi Naito 1 , Kazumi Akasaska 3 , Shuichi Kino 3 , Yutaka Tomoda 3 , Tadashi Matsubayashi 4 ,
Nobuyuki Kubota 2 , Junichi Ando 5 , Shigemi Hosogaya 6 , Yoshihisa Itoh 1 .
Asahikawa Medical University 1 , Eiken Chemical Co., Ltd. 2 , Asahikawa Medical University Hospital 3 ,
Nissui Pharmaceutical Co., Ltd. 4 , SRL, Inc. 5 , Kagawa Prefectural University of Health Sciences 6 , Japan
Aim Reference material is a key in setting reference measurement system. If it is serum-matrix base
and analytes contained are closely similar, it can be used for many other clinically relevant purposes.
We have newly developed the multi-purpose serum-matrix base reference material.
Materials and Methods The material was prepared in a similar manner as ERM-DA470, an
international reference material of serum proteins, IRMM. In short it is lyophilized after processing
normal pooled sera (Shima Laboratories, Japan), reconstituted with hepes buffer, and spiked with CRP,
cystatin C, and beta-2-microglobulin.
Results We have successfully prepared the material with high safety, short-term stability, and
long-term stability. Tests are negative for HBV, HCV, and HIV. After reconstituting with 1 ml of water
the value was unchanged at 4ºC and room temperature until 12 hours. We confirmed the stability for
one year. Storage at 4ºC and -80ºC gave a good result. These include CRP, cystatin C, beta
2-microoglobulin, pepsinogen I, II, and others. No inter-vial differences are noted. 31 different serum
proteins and cytokine value was initially set by participation of immunoassay systems. The value in
each analyte was generally 2/3 of that of normal adult, which may be caused by amount of distilled
water added. Some minute discrepancy was found in the reaction of cystatin C and beta2-microglobulin
between the material and patient’s sera.
Comments The present prepared material definitely play a unique role for standardization for
immunoassay of different kinds of analytes, especially non-standardized controls for everyday’s
laboratory services, and references of a given analyte for experimental purpose.
- 68 -

No. 16 (PC 16)
Temporal changes in estimated glomerular filtration rate (delta eGFR) may be a
predictor of progression to coronary heart disease
So-Young Kim, Tae-Dong Jeong, Woochang Lee, Sail Chun, Won-Ki Min*
Department of Laboratory Medicine, Asan Medical Center and University of Ulsan College of Medicine,
Seoul, Korea
Background: Chronic kidney disease and cardiovascular disease are major public health problems
worldwide and often share the same pathophysiological mechanisms. Indeed, the prevalence of
traditional cardiovascular risk factors can be higher in those with impaired kidney function, and most
patients with a lower estimated glomerular filtration rate (eGFR) die of cardiovascular causes and not
due to progression to end stage renal disease. However, the effect of reduced eGFR on coronary heart
disease has not been well delineated among Korean population. Here, we reviewed temporal changes in
eGFR (delta eGFR, ΔeGFR) of the healthy Korean population, and then, carried out a study to
investigate the link between a ΔeGFR and the risk of coronary heart disease.
Methods: We reviewed data of 7676 healthy outpatients selected from a population of - 10,000
individuals referred to the health promotion center of the ASAN medical center between January 2010
and December 2010. At the time of the first evaluation, data of patients` age, sex, blood pressure,
creatinine, total cholesterol, LDL, HDL and triglycerides as well as history of smoking, hypertension or
familial coronary heart disease were collected by chart review. The % risk of coronary heart disease
were calculated by Framingham score (Score:www.nhlbi.nih.gov/guidelines/cholesterol). We classified
ΔeGFR % into 4 groups according to Gaussian distribution, ±2 standard deviation (SD); below -2SD,
-2SD to mean, mean to +2SD, over +2SD (mean=1.559, SD=9.5837).
Results: Among the enrolled patients, 5.9% (456 patients) were over 15 of the Framingham score, and
this is equivalent of coronary heart disease risk over 20% which means they have high risk for
occurrence of coronary heart disease in ten yrs. A group of over +2SD (ΔeGFR > 19.17%) had odds
ratio of 3.656, and it was related to incident % risk of coronary heart disease rather than other ΔeGFR
groups.
Conclusions: A decline of eGFR > 19.17% was independently related to incident % risk of coronary
heart disease. ΔeGFR could be a possible indicator of progression to the coronary heart disease in
general population. Therefore, this parameter may be potentially used for screening subjects with higher
risk of coronary heart disease.
Keywords: estimated glomerular filtration rate (eGFR), delta eGFR (ΔeGFR), coronary heart disease,
chronic kidney disease
- 69 -

No. 17 (PC 17)
Development of C-reactive protein assay for human saliva
Yumi Narita, Taku Shirakawa
Department of Biophysics, Graduate School of Health Sciences, Kobe University, Japan
BACKGROUND: Measurement of C-reactive protein (CRP) in saliva may be able to be used for the
early detection of the inflammatory disorder of infants or home care patients. However, the reports about
the correlation between salivary and serum CRP vary among researchers. Such differences might be
caused in the step of pretreatment or collection of saliva. In this study, we examined these steps in order
to develop a high-sensitivity assay for salivary CRP.
METHODS: Salivary CRP concentration was determined by sandwich enzyme-linked immunoassay
(ELISA). To improve the sensitivity of the assay, Ampliflu TM Red (Sigma) was used as a fluorogenic
substrate. Saliva was collected by drooling and/or saliva collection swab. Saliva samples were diluted
1:1 in sample diluent containing Tween20 and/or cation, then centrifuge at 1500g for 5 minutes.
Concentrations of Tween20 and cations were examined in 0 to 1% and 0 to 1000mM, respectively. The
validity of ELISA was examined by intra-assay precision and recovery test. Saliva and serum samples
from 28 healthy volunteers were collected, and then CRP concentrations were measured in order to
evaluate the correlation of salivary and serum CRP.
RESULTS: 500mmol/L MgCl2 and 0.25%Tween20 were best at enhancing the CRP fluorescent signal.
Both MgCl2 and Tween20 were essential to detect strong signal. Under these conditions intra-assay CV
was 3.0%, and recovery was 102-107%. CRP concentration decreased 30% in samples collected with
saliva collection swabs in comparison with drooling. CRP was measurable in saliva samples (84.8 to
4104.3pg/mL) and serum samples (26.0 to 2015.4ng/mL). There was a positive correlation between
salivary and serum CRP (r=0.77).
CONCLUSIONS: Strong enhancement of CRP fluorescent signal was observed in the presence of
MgCl2 and Tween20. Salivary CRP concentration was affected by collection method. Salivary CRP and
serum CRP moderately correlated. This result will support for the validation of salivary CRP as an
alternative marker of inflammation.
- 70 -

No. 18 (PC 18)
IODINE STATUS AMONG THE SCHOOL CHILDREN OF HILLY AND PLAIN
REGION OF EASTERN NEPAL
Baral N 1 , Khatiwada S 1 , Shakya PR 1 , Sah GS 2 , Gelal B 1 , Das BKL, Lamsal M 1
1 Department of Biochemistry, 2 Department of Pediatrics and Adolescent Medicine, B. P. Koirala
Institute of Health Sciences, Dharan, Nepal
Introduction: Iodine deficiency is a leading cause of preventable mental retardation in the world today
affecting 19.4% population of Nepal. It is particularly affect the developing brain of fetus and neonates.
The geographical situation and poor accessibility of adequately iodized salt are the main cause of iodine
deficiency in our country. Urinary iodine excretion (UIE) reflects current iodine intake and salt iodine
content explore the level of iodine in household level.
Objective: To assess the iodine status of the school children in Eastern Nepal.
Materials and methods: A community based study was conducted on twelve districts of the Eastern
Nepal representing hilly (Taplejung, Panchthar, Ilam, Dhankuta, Tehrathum and Sankhuwasabha) and
plain region (Jhapa, Morang, Sunsari, Saptari, Siraha and Udayapur) on January 2008 to January 2012.
A total of 4525 primary school children (6-12 years) were enrolled in the study and equal numbers of
salt and urine samples were collected for the assessment of iodine status. UIE was measured by
ammonium-persulphate digestion microplate (APDM) method and salt iodine content by semi
quantitative rapid test kit.
Results: Our study showed that 583 (12.9%) of school consumed open salt whereas 3942 (87.1%)
consumed packet salt. Majority (86.7%) of school children consumed adequately iodized salt but 10.2%
consumed inadequately iodized salt and 1.1 % children consumed salt with no iodine. The median UIE
of the all school children is 241.08µg/L indicate the adequate iodine nutrition but still 13.1% children
were suffering from some degree of iodine deficiency.
Conclusion: Based on the result we conclude that iodine status of the school children in Eastern Nepal
is improving but continuous monitoring of the population is necessary for the sustainable elimination of
iodine deficiency disorders (IDD) from the country and prevention of iodine induced diseases.
- 71 -

No. 19 (PC 19)
Lipid Profile in Menopausal women of Different Parity and Basal Metabolic index
status in South west, Nigeria
Adesina Adeyemi Adeleke; Ogunlewe Jayeola; Oyedeji Samuel
Department of Chemical Pathology, OAUTHC, Ife, PMB5538, Osun-state, Nigeria
Background: Studies from different part of the world have reported the changes in
plasma lipid and lipoproteins cholesterol in post-menopausal women and its attendance
risk of cardiovascular disease. Many researchers are still gathering the contributing
factors to this condition. The research was carried out to evaluate some factors that may
contribute to the dyslipidemic lipid profile observed in post-menopausal women.
Methods: Analysis of fasting plasma cholesterol, lipoproteins and triglycerides were done
using standard spectrophotometric methods in eighty (80) post-menopausal women and
fifty (50) pre-menopausal women. Basal metabolic index were determine by measuring
their weight and height. Parity status of each subject was noted.
Results: Post-menopausal women lipid profile shows dyslipidemic picture compare with
pre-menopausal women. Total cholesterol 6.02±1.0 and 4.08±0.84mmol/l, P

No. 20 (PC 20)
Plasma free Fatty Acid Levels in Norma- and Hyper-tensile pregnant women before
and 72 Hour after delivery in Osun-state Nigeria
Adesina Adeyemi Adeleke; Oyedeji Samuel; Taiwan Funke
Department of Chemical Pathology, OAUTHC, Ife, PMB5538, Osun-State, Nigeria
Background: Pregnancy induces hypertension has been one of the major threats to improve
life expectancy of our women. While majority lack access to good health care facilities,
Gynecologist still find it difficult to identify a specific marker for predicting or biochemical
responsible for its pathogenesis.
Aim: Estimation of fasting Plasma level of free fatty acid in second, third trimesters, 24
hours before delivery and three days after delivery in norm and hypertensive pregnant
women.
Method: Twenty (20) hypertensive pregnant women and twenty normotensive pregnant
women attending anti-natal clinic and admitted to labor and post-natal ward were involved
in this study. The average blood pressure of hypertensive pregnant women in second, third
trimester, 24hours before delivery and 72 hours after delivery were 150/100 mmHg,
170/110mmHg and 170/120mmHg and 150/90mmHg. Normotensive pregnant women have
average of 110/70mmHg in all stages. Plasma free fatty acid was estimated using standard
spectrophotometric method.
Results: The mean plasma free fatty acid in norm and hypertensive pregnant women in
various stages are; second trimester 2.7±0.2 and 2.3±0.3mmol/l, P

No. 21 (PC 21)
Lipid Peroxidation and Selected Non-Enzymatic Antioxidants in Sickle Cell Disease
Patients in South West, Nigeria
Adesina Adeyemi Adeleke; Oyedeji Samuel; Oladepo Ridwan; Oke Taiwo; Fasakin Kolawole
Department of Chemical Pathology, OAUTHC, Ife, PMB5538, Osun-State, Nigeria
Department of Heamatology and Blood Transfusion, OAUTHC, Ife, Osun-State, Nigeria
Objective: Sickle cell anaemia (SCA) is hereditary disorder with higher potential for oxidative damage
due to chronic redox imbalance in red cells and vaso-occlusive crisis.
Methods: Levels of lipid peroxidation and vitamins antioxidant status were determine in fifty (50)
sickle cell anaemia patients in Osun State, Nigeria. Thirty (30) normal heamoglobin individual were
used as controls. Malondialdehyde (MDA), Vitamin C, vitamin E, Lipid profile were estimated in
patients and control serum using standard spectrophotometric analysis.
Results: The results showed significant increase in MDA of patients compared with control 0.94± 0.2
and 0.49±4µmol/l, P

No. 22 (PE 1)
Two-Step Biochemical Differential Diagnosis of Classic 21-hydroxylase Deficiency
and Cytochrome P450 Oxidoreductase Deficiency in Japanese Infants Using
Urinary Steroid Metabolites by Gas Chromatography - Mass Spectrometry
Yuhei Koyama 1)2) , Keiko Homma 3) , Maki Fukami 4) , Masayuki Miwa 5) , Kazushige Ikeda 5) ,
Tsutomu Ogata 4) , Tomonobu Hasegawa 5) , Mitsuru Murata 1)
1) Department of Laboratory Medicine, Keio University School of Medicine, 2) Mitsubishi Chemical
Medience Co., 3) Central Clinical Laboratories, Keio University Hospital, 4) Department of
Endocrinology and Metabolism, National Research Institute for Child Health and Development, 5)
Department of Pediatrics, Keio University School of Medicine, Japan
[Introduction] The clinical differential diagnosis of classic 21-hydroxylase deficiency (C21OHD) and
cytochrome P450 oxidoreductase deficiency (PORD) is sometimes difficult since both deficiencies can
have similar phenotypes and high blood concentrations of 17α-hydroxyprogesterone (17OHP). The
objective of this study was to explore novel biochemical indexes for the differential diagnosis of
C21OHD, PORD, and transient hyper 17α-hydroxyprogesteronemia (TH17OHP) in Japanese newborns.
[Material and Methods] We recruited 29 infants with C21OHD, 9 infants with PORD, 67 infants with
TH17OHP, and 1341 control infants. All infants were Japanese between 0-180 days old. None
received glucocorticoid treatment before urine sampling. We measured urinary pregnanetriolone (Ptl),
the cortisol metabolites 5α- and 5β-tetrahydrocortisone (sum of these metabolites defined THEs), and
metabolites of three steroids, namely dehydroepiandrosterone, androstenedione (AD4) and
11β-hydroxyandrostenedione (11OHAD4) by gas chromatography-mass spectrometry.
[Results] By using a cutoff 0.020, the ratio of Ptl to THEs, we were able to differentiate C21OHD and
PORD from TH17OHP and controls with no overlap (minimum – maximum: 0.14-15, 0.051-0.23,
0.00010-0.011, and 0.000068-0.0083, respectively). Among metabolites of DHEA, AD4, and
11OHAD4, only 11β-hydroxyandrosterone (11HA), a metabolite of 11OHAD4, showed no overlap
between C21OHD and PORD (0.61-24 and 0.022-0.22 mg/g creatinine, respectively) using a cutoff of
0.35.
[Conclusion] We successfully established a 2-step biochemical differential diagnosis for C21OHD and
PORD by urinary steroid metabolites such as Ptl, THEs, and 11HA.
- 75 -

No. 23 (PE 2)
Total cholesterol/HDL cholesterol ratio and LDL cholesterol/Apo B ratio as risk
factors for cardiovascular disease in prediabetic of first-degree relatives from
patients with type 2 diabetes mellitus.
Bety Kus Rini Sari, Marzuki Suryaatmadja
Department of Clinical Pathology, Medical Faculty, Universitas Indonesia
Prediabetic patients can progress to type 2 diabetes mellitus (t2DM) and cardiovascular disease
(CVD). Lipid profile has an important role in CVD. However there is still few data about Apo B level,
total cholesterol/HDL-cholesterol ratio (TC/HDL-C ratio) and LDL-cholesterol /Apo B ratio
(LDL-C/ApoB ratio) in first-degree relatives from patients with t2DM in Indonesia. This study aimed
to get picture of Apo B level, TC/HDL-C and LDL-C/Apo B ratios in the first degree relatives from
patients with t2DM as markers for early detection of CVD.
The prediabetes i.e. impaired fasting glucose or impaired glucose tolerance was diagnosed based on
blood glucose level. This cross sectional designed study consisted of 56 prediabetic subjects and 56
normoglycaemic subjects, all were the first-degree relatives from patients with t2DM. Total cholesterol,
HDL cholesterol, LDL cholesterol were determined by enzymatic and direct homogenous enzymatic
methods whereas Apo B was determined by immunoturbidimetric methods.
In prediabetic subgroup TC/HDL-C ratio was significantly higher (4.5±1.1 vs 3.2± 0.5; p

No. 24 (PE 3)
Adiponectin HMW Level and Homeostasis Model Assessment-Insulin Resistance
(HOMA-IR) in Type 2 Diabetes Mellitus First Degree Relatives
Yulia Sari, Marzuki Suryaatmadja, Budiman Darmowidjojo
Department of Clinical Pathology, Medical Faculty, Universitas Indonesia
Type 2 diabetes mellitus (t2DM) accompanied by macro and microangiopathy complications had
been a serious global problem, with an increasing morbidity and mortality rates.
The most important pathogenesis is insulin resistance (IR), which could be predicted by the
Homeostasis Model Assessment-Insulin Resistance (HOMA-IR). Adiponectin has been
already known to increase the insulin sensitivity and play role in balancing the
glucose metabolism. It is very important to be able to detect the prediabetes early to prevent the
complications.. The aim of this study is to get the picture of adiponectin HMW level and the HOMA-IR
as IR predictors in the t2DM first degree relatives, and the correlation between adiponectin level with
fasting insulin and HOMA-IR. This cross-sectional designed study was conducted from April 2011 until
January 2012, on t2DM first degree relatives. The subjects consisted of 52 prediabetes and 52
normoglycaemic respondents. The result revealed that in prediabetic group, the adiponectin HMW
levels in both men and women were significantly lower (P=0,003), HOMA-IR were significantly higher
than in the normoglycaemic group (P=0,0026), and there were negative correlations
between adiponectin HMW level in both men and women with HOMA-IR and fasting
insulin. These findings had proven that IR had been presented in prediabetes stage which was correlate
well to the hypoadiponectinemia. Based on these results, it is recommended to measure adiponectin
HMW, insulin and HOMA-IR in t2DM first degree relatives to detect the IR and to prevent the sickness
earlier to decrease the morbidity and mortality rates.
Keywords: Type 2 Diabetes Mellitus, first degree relatives, prediabetes, insulin resistance,
HOMA-IR, adiponectin HMW
- 77 -

No. 25 (PE 4)
Age-related Anti-Müllerian hormone Concentrations in Healthy Korean Females
Lee, A 1 ; Jang, TG 2 ; Lim, SY 2 ; Park, JC 2 ; Rhee, JH 2 ;Kim, YJ 1 ; Lee, KY 1
Seoul Medical Science Institute/Seoul Clinical Laboratories, Seoul, Korea 1 , Department of Obstetrics
and Gynecology, School of Medicine, Keimyung University, Daegu, Korea 2
Backgrounds : Currently, serum anti-Müllerian hormone (AMH) in women has been increasingly used
for ovarian reserve function assessment in fertility centers. Here we evaluated age-related AMH values
in various ages of healthy females in Korea, and investigated the relationships with antral follicle count
(AFC) and other fertility hormones.
Methods: We recruited 222 healthy females without specific infertility history in various ages. For
women of reproductive age, blood samples were drawn at day 3 from the onset of cycle. AMH was
measured by ELISA (Immunotech SAS, France). E2, FSH and LH were measured by
chemiluminescence immunoassay (ADVIA Centaur ® XP, Siemens, USA). Statistical analysis was
performed using MedCalc software (version 12.3, Belgium). AMH values were calculated in different
age groups, and correlation of AMH and other hormones were analyzed. For 104 patients whose AFC
assessment was available, correlations of AFC and each hormone were also analyzed.
Results: 2.5 - 97.5 percentiles of AMH values depending age groups are as follows: 0.10 - 3.99 ng/mL
(60 years, n=43). AMH negatively correlated with FSH (r =
-0.4213, P < 0.0001), LH (r = -0.3858, P < 0.0001) and E2 (r = -0.2070, P = 0.0025). Regarding the
relationship of each hormone with AFC, AMH positively correlated with AFC (r = 0.7375, P < 0.0001),
while negatively correlated FSH (r = -0.5401, P < 0.0001).
Conclusion: AMH values should be interpreted in caution, with regard to women’s age, because AMH
values are steadily increased from birth until 20 th , and continuously decreased after 30 th . AMH best
correlates with AFC, which suggest AMH is one of the most promising markers for assessment of
ovarian reserve function.
- 78 -

No. 26 (PE 5)
Analysis of von Willebrand Factors’ Level in Type 2 Diabetes Mellitus
Mansyur Arif 1 , Ichwan Meinardi 1 , Uleng Bahrun 1 , Harsinen Sanusi 2
1 2
Department of Clinical Pathology, Department of Internal Medicine,
Faculty of Medicine Hasanuddin University, Makassar, Indonesia
The aim of this study was to analyze von Willebrand Factor (vWF) level as the risk factor of thrombosis
in type 2 diabetes mellitus. A cross sectional study had been done at Endocrine Metabolic Subdivision
Department of Internal Medicine of Wahidin Sudirohusodo Hospital. Laboratory examination had been
done at Clinical Pathology Laboratory of Wahidin Sudirohusodo Hospital Makassar. Seventy-five
participants consisted of 52 subjects with type 2 DM and 25 normal subjects as controls had been
examined. The results of this study showed that the age of type 2 DM subjects were ranging from 39-74
years old and the age of normal controls were ranging from 30-44 years old. The level of vWF in type 2
DM and normal controls were 86-362% and 41-210%, respectively. There was significantly increased of
vWF level in type 2 DM. It was proven that in subject with type 2 DM occurred hypercoagulation states
that characterized by elevated level of vWF. The level of vWF is suggested to be a marker of
hypercoagulation states caused by endothelial dysfunction in type 2 DM. Further study is suggested to
determine the level of vWF in type 2 DM by including other risk factors such as hypertension,
dyslipidemia, smoking and obesity.
Key words: Type 2 Diabetes mellitus, von willebrand factor
- 79 -

No. 27 (PI 1)
Combination of Anti-HCV Assay to Improve Performance of Hepatitis C Screening
Test in Prodia National Reference Laboratory, Indonesia
Romimatul Fadhilah, Yenny Surjawan, Indriyanti R. Sukmawati
Prodia Clinical Laboratory, Indonesia
Anti-HCV assay for hepatitis C screening test has been widely used. Based on CDC guideline (2003),
positive anti-HCV screening result has to be verified by immunoblotting assay or nucleic acid testing. In
fact, not all laboratories in Indonesia have facilities to perform this test. Prodia National Reference
Laboratory received samples for anti-HCV screening test from its hundred branches. Samples sources
were from vary population with varying prevalence of hepatitis C. In our experience, samples referred
for anti-HCV assay from other laboratories quite often showed discordance results with ours. Previous
evaluation of 4 automated analyzers for anti-HCV assay by Kim et al (2008) showed good sensitivity
(100 %), but 3.7 % still showed false positive results. Therefore an evaluation of routine anti-HCV
screening test should be performed. We evaluated 55 samples using 3 analyzers (Advia Centaur,
Architect-i2000 and Axsym). Seventeen samples that showed discordant results among analyzers or had
extreme low and high index or s/co were confirmed by immunoblotting assay using INNO-LIA TM HCV
Score reagent (Innogenetics). The sensitivity of anti-HCV assay on those 3 analyzers was good (100 %).
False positive results were found in 4 samples by Architect, 5 samples by Advia and 5 samples by
Axsym. The assay showed a different result if a sample, especially sample with low s/co ratio or index,
was examined with different analyzers. The false positive results were not found in the same samples
and there were no specific pattern. Therefore, we consider using a second anti-HCV assay as
recommended by UK Health Protection Agency algorithm (2007) for results with low s/co ratio or index
to reduce the possibility of false positive result.
- 80 -

No. 28 (PI 2)
Diagnostic value of AFP-L3 and PIVKA-II in hepatocellular carcinoma according to
the level of total-AFP
Eun-Jee Oh 1 , Seung Won Jung 1 , Jong Young Choi 2 , Hee Yeon Kim 2 , Myungshin Kim 1 , Yonggoo
Kim 1 , Dong Goo Kim 3
Department of Laboratory Medicine 1 , Internal Medicine 2 and Surgery 3 , School of Medicine, The
Catholic University of Korea, Korea
AIM: Although, alpha-fetoprotein (AFP) is the most widely used tumor marker for hepatocellular
carcinoma (HCC), AFP lacks adequate sensitivity and specificity for effective surveillance. The aim of
this study was to evaluate diagnostic value of AFP-L3 and Prothrombin induced by vitamin K
absence-II (PIVKA-II) in HCC.
METHODS: One hundred and sixty-eight patients (90 HCC, 78 benign liver diseases) during routine
HCC surveillance were included. Sera were obtained during their first evaluation for HCC development
and at the time of HCC diagnosis before commencing HCC treatment. AFP, AFP-L3 and PIVKA-II
were measured in the same serum by microchip capillary electrophoresis and liquid-phase binding assay
on a μTAS Wako i30 auto analyzer
RESULTS: In a total of 168 patients, areas under the curve (AUC) for HCC were 0.879, 0.887, 0.801
and 0.939 for AFP, AFP-L3, PIVKA-II and the combined markers, respectively. AFP-L3 had higher
AUC than PIVKA-II for HCC (P = 0.043). In 112 patients with low AFP levels (5%) and PIVKA-II (cut-off >40AU/L), the sensitivities were 92.1-94.4% and
specificities were 75.6-79.7% in all patients and subgroup. Combined markers detected about 90% of
early stage, small sized or single tumors in the low AFP group. In multivariate analysis, AFP-L3 was
correlated with AFP and tumor size, and PIVKA-II was correlated with laboratory tests including serum
AST, total bilirubin, platelets and albumin levels. PIVKA-II had no correlation with AFP, AFP-L3 or
tumor characteristics.
CONCLUSION: Combined determination of AFP-L3 and PIVKA-II could improve the diagnostic
value for HCC detection in patients with or without increased AFP levels. The utility of improved
surveillance protocol based on these tumor markers needs to be investigated.
- 81 -

No. 29 (PI 3)
Development of ABO and Lewis typing by enzyme-linked immunosorbent assay for
disaster
Terutaka Sagawa and Mariko Okada
Division of Biomedical Sciences, Institute of Medical Technology, Ehime Prefectural University Of
Health Sciences, Japan
For ABO blood group typing, we need electrical power, the clean area for collecting venous blood and
substantial experience of technologists. We have little electrical power, clean areas and human resources
after a disaster, just like a giant earthquake or a giant tsunami, occurs. In this case, there is a need of the
method that does not require electrical power, the clean area and substantial experience.
We have developed ABO and Lewis blood group typing by enzyme-linked immunosorbent assay
( ELISA ) using Japanese saliva as antigen. Tween 20 was used to block and wash. This method could
give a result within 120 minutes from saliva collecting procedure.
All secretors (88% of the samples examined) reacted with anti-H Ab.
Nonsecretors reacted weakly with anti-H Ab or not. A-type and AB-type nonsecretors reacted weakly
with anti-A Ab. B-type and AB-type nonsecretors reacted weakly with anti-B polyclonal antibody but
not anti-B monoclonal antibody.
Secretors included Lea-Leb- (19% of secretors), Lea-Leb+ (28%) and Lea+Leb+ (53%). All
nonsecretors reacted with anti-Lea Ab not anti-Leb Ab.
Kudo et al. demonstrated that almost Japanese nonsecretors had sej/sej homozygosity. The sej/sej has
an elevated ABH antigen. No activity alleles are very rare in Japanese.
It was proved that the ELISA method reported here was useful for almost Japanese ABO and Lewis
blood group typing, and did not require electrical power and substantial experience. Further
developments are necessary for very rare Japanese complete nosecretors, and Rh typing and ABO
reverse typing.
- 82 -

No. 30 (PI 4)
Evaluation of Hepatrio multiplex kit for early and simultaneous detection of
hepatitis A, B and C
Chi Hyun Cho, M.D. 1 , Sang Kyung Jo, M.D. 2 , Young Joo Kwon, M.D. 2 , Chae Seung Lim, M.D. 1
and Yun Jung Cho, M.D. 1*
1
Department of Laboratory Medicine, College of Medicine, Korea University, Seoul, Korea
2
Department of Nephrology and Hypertension, College of Medicine, Korea University, Seoul, Korea
Introduction: In hemodialysis patients, early detection of hepatitis A, B and C is very important. While
serology test for detection of hepatitis A, B and C is used, it has limitation in that it does not catch the
early infection and co-infection of hepatitis A, B and C. Recently a multiplex PCR kit for the
simultaneous detection of hepatitis A, B and C, Hepatrio, was developed. We evaluated the multiplex kit
in hemodialysis patients in one Korea hospital and compared the result with serology.
Methods: HAV IgM, HBsAg, anti-HBs, anti-HBc and HCV Ab were analyzed on 186 samples of
routine check-up examinees in a hemodialysis care unit by chemiluminescence immunoassay. Nucleic
acid amplification test (NAT) was performed on the same samples by multiplex real-time polymerase
chain reaction (PCR) kit for simultaneous detection of HAV, HBV, and HCV (Magicplex Hepatrio;
Seegene, Seoul, South Korea).
Results: The positive rates of HAV, HBV, and HCV were 0, 5.9, and 0.6%, respectively. All samples
except two showed consistent results between serology and multiplex PCR. Occult HCV infection was
found in 1 (0.6%) of 173 HCV-Ab negative subjects. Meanwhile, a HBs-Ag positive sample was not
detected for HBV-DNA by multiplex PCR. There was no co-infection case among samples.
Conclusion: This study suggests the possibility that multiplex PCR replaces serology for screening of
hepatitis A, B and C. Further study using much more samples is needed for the clinical performance
evaluation of multiplex PCR test. Key word: Hepatitis A, hepatitis B, hepatitis C, hemodialysis,
serology, multiplex PCR kit
- 83 -

No. 31 (PI 5)
Impact of interleukin-29 on interferon alpha secretion of plasmacytoid dendritic cell
Chi Hyun Cho, M.D. 1 , Chae Seung Lim, M.D. and Yun Jung Cho, M.D. 1*
1 Department of Laboratory Medicine, College of Medicine, Korea University, Seoul, Korea
Background: The alpha-IFN secretion by plasmacytoid dendritic cell was distinguished by rapid
kinetics, high level (up to 1000 fold higher than most cells), while the plasmacytoid dendritic cells
responded with vigorous alpha-IFN production to stimulation by a variety of viral stimuli and to
synthetic Toll-like receptor agonists. IL-28Rα (type III IFN receptor chain) signaling was demonstrated
not to provide feedback on type I expression in mouse study. In this study, we tried to identify whether
the type III IFN provides feedback or does not do so on alpha-IFN secretion of human plasmacytoid
dendritic cell.
Methods: From eleven healthy donors, 3 ml and 10 ml of whole blood were collected for complete
blood cell count by Coulter automatic analyzer and for the isolation of peripheral blood mononuclear
cells (PBMC). Plasmacytoid dendritic cell was counted by multiplying % of total PBMC
(flowcytometer) by mononuclear cells of whole blood (Coulter automatic analyzer). PBMC from each
healthy donor was then divided into four groups as follows: group 1- control; group 2- addition of CpG
DNA; group 3- addition of IL-29; group 4- addition of CpG DNA and IL29. Then, the culture media
were incubated for 24 hours at 37℃. The supernatants were collected and tested for the amount of
IFN-alpha secretion by the flowcytometer.
Results: The number of plasmacytoid dendritic cells (/µL) was 5.29 (±3.05). IFN-alpha (pg/ml)/pDC
(/µL) in each group was as follows: 5.677 (±9.278) in group 1; 1071.546 (±1026.598) in group 2;
14.142 (±21.107) in group 3; 1913.897 (±1525.928) in group 4. There were statistically significant
differences between group 1 vs 2 and group 2 vs 4, whereas there was no statistically significant
difference between group 1 vs 3. The addition of CpG DNA with IL-29 caused human pDC to secret
IFN-alpha about two times more than addition of CpG DNA alone.
Conclusions: Apart from mouse study, one of lambda-IFNs, IL-29 provided feedback on IFN-alpha
expression of human pDC. IL-29 alone did not have impact on IFN-alpha secretion by human pDC, but
with CpG DNA, had synergistic effect on that. Given the knowledge that IL-29 is potential therapeutic
alternatives to type I IFNs in terms of viral infections and tumors, this result could help elucidate the
mechanism of the therapeutic effect by IL-29.
Key Words : CpG DNA, IL-29, IFN-alpha, plasmacytoid dendritic cell
- 84 -

No. 32 (PI 6)
Discrepancy between serologic and genotyping results of ‘Mi a ’ blood antigen and
antiserum in Taiwan
Chien-Feng Sun 1,2 , Tai-Di Chen 2 , Ding-Ping Chen 2
1 Department of Anatomic Pathology, 2 Department of Laboratory Medicine, Linkou Medical Center,
Chang Gung Memorial Hospital, Taoyuan, Taiwan
Aim. Mi.Ⅲ (GP.Mur) is reported to have a mean frequency of 7.3% among Taiwanese. Since the
specificities of Mi.III (GP.Mur) antisera are not further specified, such antisera and the cells identified
by them have been applied a collective term as anti-‘Mi a ’ and ‘Mi a ’(+) cells in Taiwan. Anti-‘Mi a ’
antibodies are the most common clinically important antibodies detected in Taiwan. Our aim of this
study is to evaluate the efficiency of current practice of using ‘Mi a ’(+) screening cells in comparing to
genotyping for Miltenberger series.
Materials and Methods. Blood samples were obtained from randomly selected patients. Antisera
containing anti-‘Mi a ’ specificity were collected routinely in our blood bank. Manual polybrene method
without a supplementary antiglobulin phase was used for serologic test. The results of serologic test
were compared with PCR-sequencing data. The study was approved by our Institutional Review Board.
Results. The prevalence of Mi.Ⅲ(GP.Mur) is 6.4% (25/389). Among 1945 serologic tests for these
389 samples, 210 tests showed a positive reaction (1+ or more). Among them, only 84 positive results
belonged to PCR (+) samples. The sensitivity and specificity of our routine practice using ‘Mi a ’(+)
screening cells to detect anti-‘Mi a ’ antibodies are thus estimated to 67.2% (84/125) and 93.1%
(1694/1820), respectively. The positive predictive value is 40% (84/210).
Conclusions. Our results showed that the Mi.III (GP.Mur) has a prevalence of 6.4% in Taiwan, and no
GP.Hut, GP.Hop, GP.Bun, or GP.HF was detected. However, our study also showed that our screening
system for detecting anti-‘Mi a ’ has a low sensitivity of 67.2% with a specificity of 93.1%. Since a
previous study suggested the presence of other glycophorin variants in Taiwan, the results suggested the
existence of other (Mi) phenotypes and the current detecting method may not be sufficient to identify
antibodies other than anti-GP. Mur.
- 85 -

No. 33 (PI 7)
Immune response to MSP2 antigen of plasmodium falciparum in mamuji, west
Sulawesi, Indonesia
Nurhayana Sennang 1 , Dianawaty Amiruddin 2 , Bahrani 2 , Sitti Wahyuni 2 , Syafruddin 2,4 , Irawan
Yusuf 3 , Stephen Rogerson 5
1 2 3
Clinical Pathology Department, Parasitology Department, Physiology Department, Faculty of
Medicine, Hasanuddin University, Makassar, South Sulawesi, Indonesia; 4 Eijkman Institute, Jakarta,
Indonesia; 5 Department of Medicine, University of Melbourne, Post Office Royal Melbourne Hospital,
Melbourne, Victoria, Australia
Background: A malaria parasite antigen, merozoite surface protein 2 (MSP-2), is assumed to be
involved in initial attachment of merozoite to erythrocyte surface. MSP-2 may contribute to the
development of clinically protective immunity to malaria which is dependent upon cumulative exposure
to the many parasite variants circulating in the local population. The aim of this study is to determine
immune response to MSP-2 P.falciparum in individuals living in Mamuju, an epidemic area of malaria
in West Sulawesi, Indonesia. Methods: The cross sectional study was conducted using multi-stages
random sampling from 15 districts (123 villages) in Mamuju, West Sulawesi, Indonesia. Capillary blood
samples from 4406 participants were collected during August 2010. All positive samples and 5%
negative samples based on blood smear results were selected randomly for immunological investigation.
IgG antibodies to MSP-2 were analyzed by Enzyme Linked ImmunoSorbent Assay (ELISA) Results:
Of 209 samples based on blood smear results, there were 18 samples with P.falciparum parasitaemia, 10
samples with P.vivax parasitaemia and 181 negatives samples, Means of IgG antibody to MSP2 in
individuals who had positive P.falciparum blood smears was 37.54 units, higher than positive P.vivax
(7.48 units) and negative blood smears (2.61 units), there was a significant difference between the three
groups (95%CI: p=0.0001). Cut off point of IgG antibody to MSP2 was 4.57 units. Of 37 samples with
levels of IgG antibody to MSP2 more than cut off, 11 were from individuals with P.falciparum
parasitaemia, 3 from individuals with P.vivax, and 23 from uninfected people. Conclusion: Increased
level of IgG antibody to MSP2 was associated with infection with P.falciparum. The presence of IgG
antibody to MSP2 in individuals with P.vivax infection or negative blood smears may indicate
mix-infection of P.falciparum undetected by blood smear or previous infection with P.falciparum.
Keywords: immune response, MSP-2, Plasmodium falciparum
- 86 -

No. 34 (PI 8)
Inhibition of the NF-kappa B pathway as a candidate strategy for treatment of
cryopyrin-associated periodic syndrome
Satoka Takahashi 1,2 , Tetsuo Kubota 1
1 Department of Microbiology and Immunology, Tokyo Medical and Dental University Graduate
School of Health Care Sciences, Tokyo, Japan.
2 Department of Health and Science, Faculty of Health and Medical Care, Saitama Medical University,
Saitama, Japan.
Cryopyrin-associated periodic syndrome (CAPS) is caused by a mutation in the CIAS1 gene coding for
NLRP3. Once activated by pathogen-associated molecular patterns (PAMPs) and/or damage-associated
molecular patterns (DAMPs), NLRP3 forms a protein complex referred to as an inflammasome, leading
to extracellular release of IL-1beta. Since mutated NLRP3 forms an inflammasome without obvious
stimulation leading to the unrestricted release of IL-1beta, this cytokine represents a critical target for
CAPS treatment. This study aimed to test the effect of an NF-kappa B inhibitor DHMEQ (provided by
Dr Umezawa, Aichi Medical University) upon IL-1beta production, as well as the expression of other
proinflammatory molecules induced by IL-1beta.
Human umbilical vein endothelial cells (HUVECs) and peripheral blood mononuclear cells (PBMCs)
were stimulated and mRNA expression of proinflammatory proteins was assessed by quantitative
RT-PCR. The effects of DHMEQ were tested in parallel with anakinra, an IL-1 receptor antagonist.
Under stimulation by IL-1beta, mRNA expression for IL-1beta, TNF alpha, IL-6 and VCAM-1 in
HUVECs was markedly increased, and this effect was almost completely suppressed by both DHMEQ
and anakinra. However, when PBMCs were stimulated with IL-1beta, anakinra was less effective,
possibly because cells were additionally stimulated by the secondary action of cytokines other than
IL-1beta. Enhanced expression of IL-1beta mRNA in PBMCs stimulated with LPS alone, or LPS plus
ATP, was effectively suppressed by DHMEQ but not by anakinra.
In an assay system that mimics PAMPs and/or DAMPs induced IL-1beta production, DHMEQ
effectively suppressed IL-1beta mRNA as well as other IL-1beta-induced mRNA of proinflammatory
proteins, suggesting that DHMEQ is a good candidate agent for the treatment of CAPS.
- 87 -

No. 35 (PH 1)
Gene Diagnoses of Tumors of Hematopoietic Organs - Diagnosis of Leukemia by
both Southern Method and PCR Method –
Kazuhiro Kabe
Business Department of Medical Technology, Laboratory of Medical Technology
Medic21 Association, Japan
Aim : I introduced a genetic test for a diagnosis of lymphocytic leukemia that was one of tumors of
hematopoietic organs. I detected the reconstituted genes of both Immunoglobulin-Heavy-Chain (IgH)
and T-Cell-Receptor (TCR) with both Southern Method and PCR Method. And I made a trial of the
diagnostic test of lymphomas of both B cell tumors and T cell tumors.
Materials and Methods : There are V (V), D (D) and J (J) regions in IgH genes. I used Southern
Method by probes to J and Semi-Nested PCR Method by primers to both V and J so as to detect the
special unit (CDRⅢ) after IgH reconstitution. There are V (Vδ), D (Dδ) and J (Jδ) regions in TCR
genes. I used Nested PCR Method by primers to Vδ, to Jδ and to the outside of both Vδ and Jδ so as to
detect the special unit (Vδ1･D･Jδ1) after TCR reconstitution.
Results : I could confirm the IgH reconstitution by Southern Method at the obvious cellular specimen
of B cell leukemia and the TCR reconstitution by PCR Method at the obvious cellular specimen of T
cell leukemia. As for the PCR limit, I could detect till the specimen that was 1/1000 as dilute as DNA
(1µg) at IgH reconstitution and till the specimen that was 1/10000 as dilute as DNA (1µg) at TCR
reconstitution.
Conclusions : Southern Method is not sensitive enough and needs much DNA (10µg) and takes too
much time for its work. PCR Method is highly sensitive and sufficient in small DNA (1µg) and this
work is practicable in one day. As PCR Method is effective to operate and we seem to be able to expect
the genetic test detect from peripheral blood.
- 88 -

No. 36 (PH 2)
Identification of autoantibodies expressed in acquired aplastic anaemia
Kageaki Kuribayashi 1,2 , Maki Goto 1 , Yusuke Takahashi 1,2 , Takashi Kondoh 1,2 , Maki Tanaka 1,2 ,
Daisuke Kobayashi 1,2 , Naoki Watanabe 1,2,
1 Department of Clinical Laboratory Medicine, Sapporo Medical University School of Medicine, Japan
2 Division of Laboratory Diagnosis, Sapporo Medical University Hospital, Japan
Acquired aplastic anaemia (AA) is recognized as an autoimmune disorder; however, the autoantigens
and target cells involved remain elusive.
Sera were obtained from acquired AA patients and the expression of autoantibodies and their target cells
were examined using the haematopoietic cell line K562 and bone marrow stromal cell line hTS-5 by
flow cytometric analysis; 43.5% and 21.7% of AA expressed autoantibody against K562 and hTS-5
cells, respectively. A cDNA library was constructed using K562 mRNA and 8 autoantigens were
identified by serological identification of antigens through recombinant cDNA expression cloning
(SEREX). Expression level of anti-CLIC1, HSPB11 and RPS27 IgG-type autoantibodies were higher in
patients with acquired AA than in normal volunteers. Moreover, 12 of 13 responders to
immunosuppressive therapy expressed at least one IgG-type autoantibody against CLIC1, HSPB11 or
RPS27, whereas all non-responders did not express any IgG-type autoantibody. Furthermore, higher
titres of anti-CLIC1 and RPS27 IgG-type autoantibodies were found to be associated with better
response rates to immunosuppressive therapy. The results of this study indicate haematopoietic cells are
the targets of immune abnormality in acquired AA. These autoantibodies may be utilized to abstract
patients associated with immune abnormality from bone marrow failure syndrome.
- 89 -

No. 37 (PH 3)
Clinical significance of overexpressed serine-threonine tyrosine kinase 1 in leukemia
Daisuke Kobayashi, Takashi Kondoh, Maki Tanaka, Kageaki Kuribayashi Naoki Watanabe
Department of Clinical Laboratory Medicine, Sapporo Medical University, School of Medicine, Japan
Objectives: Inhibitors against several tyrosine-kinases have been used in leukemia. However,
therapeutic effect is somewhat limited by drug-resistance, and therefore it is urgent to seek for novel
therapeutic targets. In this study, we analyzed the expression profile and clinical significance of
STYK1, a unique tyrosine kinase promoting cell proliferation without ligand binding, in various
leukemic patients.
Patients and Methods: In peripheral blood cells from nonleukemic group (n=15), acute leukemia (AL)
patients (n=26), and chronic myelogenous leukemia (CML) patients (n=19), STYK1 mRNA expression
was analyzed by quantitative RT-PCR. Several small inhibitory RNA (siRNA) against STYK1 and
non-silencing control RNA (NSC) were transduced by electroporation method using Nucleofector V
and Nucleofector II (Amaxa).
Results: 1) Mean measured value of STYK1 mRNA in AL and CML patients was 1,654±1,603 and
477±388, respectively, resulting 7.9-fold and 2.3-fold increase compared to the expression in
nonleukemic group (210±110). When appropriate cutoff was set using the values in nonleukemic
individuals, positive STYK1 mRNA expression was detected in 76.9% and 42.1% of AL and CML
patients, respectively. 2) STYK1 mRNA expression began to decrease after chemotherapy in AL
patients, and in the patients showing resistance to therapy, STYK1 mRNA expression preceding therapy
was significantly higher than in the patients with complete remission. 3) In the CML patients, no
relationship was found between mRNA expression level and the copy numbers of bcr-abl gene
(r=-0.21). 4) Transduction of siRNA into K562 cells with high STYK1 expression increased
doxorubicin sensitivity compared to the cells transduced with NSC.
Conclusion: These results indicate that STYK1 mRNA is highly expressed in the patients with
several leukemic patients and suggest an important role of STYK1 in the drug- resistance of acute
leukemic cells.
- 90 -

No. 38 (PH 4)
Contribution of uncontrolled fibrinolysis to bleeding diathesis in aortic aneurysm
Mitsuhiro Uchiba, Yuji Yonemura, Yukio Ando.
Department of Blood Transfusion and Cell Therapy, Kumamoto University Hospital
Kumamoto, Japan.
[Background] Disseminated intravascular coagulation (DIC) is one of the severe complications
observed in patients with aneurysm (AA). DIC results consumption coagulopathy following bleeding
diathesis. Some investigators believe that hypofibrinogenemia resulted from consumption coagulopathy
is a main cause of abnormal bleeding in AA. Bleeding diathesis is also caused by fibrinolytic
abnormality, which is often complicated with AA. To determine whether hypofibrinogenemia and
fibrinolytic abnormality contribute to bleeding diathesis observed in AA, we analyzed relationship
between bleeding symptoms and plasma levels of fibrinogen and a2-antiplasmin (a2-AP), an important
regulator of fibrinolysis. [Method] Plasma levels of fibrinogen and a2-AP in AA patients with and
without bleeding were retrospectively analyzed. [Result] Levels of both fibrinogen and a2-AP were
lower in AA patients with bleeding than in those without bleeding. Levels of a2-AP in AA patients with
bleeding were lower than reference range, whereas that of fibrinogen in bleeding patients were within
reference range. [Discussion] Our results indicated that a2-AP deficiency rather than
hypofibrinogenemia contributed to bleeding in AA. Precise mechanisms were not fully elucidated,
a2-AP deficiency resulting uncontrolled fibrinolysis may play a role. Although some investigators
recommend normalizing fibrinogen levels by using fibrinogen concentrate instead of fresh frozen
plasma in AA, our result indicated that maintaining the a2-AP level, but not fibrinogen levels, is
important. Therefore FFP but not fibrinogen concentrate should use to controlling abnormal bleeding
observed in patients with AA.
- 91 -

No. 39 (PH 5)
Aberrant DNA methylation of HOXA4 is associated with resistance to imatinib
mesylate in chronic myeloid leukemia patients
Ravindran Ankathil 1 , Marjanu Hikmah Elias 1 , Azlan H 2 , Sarina Sulong 1 , RoslineHassan 3 , Goh Ai
Sim 4 , S Fadilah Abdul Wahid 5 , Abdul Aziz Baba 2
1 2 3
Human Genome Centre, Haemato-Oncology Unit, Department of Internal Medicine, Hematology
Department, School of Medical Sciences, Health Campus, Universiti Sains Malaysia 4 Hospital Pulau
Pinang, Malaysia, 5 Medicine Department & Cell Therapy Centre, UKM Medical Centre, Malaysia
Development of resistance to Imatinib mesylate (IM) among a significant number of chronic myeloid
leukemia (CML) patients has emerged as a major clinical problem. But, mutation analysis of our CML
patients showed that BCR ABL mutations accounted for IM resistance in only 21.7% of CML patients,
indicating involvement of other BCR-ABL independent genetic and epigenetic mechanisms in
mediating resistance. We hypothesized that promoter hypermethylation of HOXA4, a gene involved in
CML progression to blast crisis, could be an epigenetic mechanism mediating IM resistance in CML
patients. Our objective was to determine the promoter hypermethylation status of HOXA4 in CML
patients on IM treatment and to evaluate its role in mediating resistance. Genomic DNA was extracted
from peripheral blood samples of 95 CML patients on IM therapy (38 good responders and 57 resistant)
and 12 normal controls, followed by bisulfite treatment. Subsequently, Methylation Specific High
Resolution Melt Analysis (MS-HRM) was performed to quantitate the methylation level of 9 CpG site,
located at -510 to -396 from the stop codon of HOXA4 gene. Five samples, representing each group of
methylation level were selected for validation using pyrosequencing. Pyrosequencing results confirmed
the methylation homogeneity of all the 9 CpG (mean SD is 4.7) in the HOXA4 promoter and their mean
methylation percentage was concordant to the MS-HRM results. Compared to good responders, HOXA4
hypermethylation level was significantly higher (P=0.002) in IM resistant CML patients. On evaluating
the risk, HOXA4 hypermethylation was associated with higher risk for IM resistance (OR 4.658; 95% CI,
1.673-12.971; P=0.003). These results prompt us to suggest that promoter hypermethylation of HOXA4
gene could be an epigenetic mechanism mediating IM resistance in CML patients and could be
considered as a potential epigenetic biomarker in addition to the BCR-ABL gene mutation, for
prediction of CML patients’ response to IM treatment.
- 92 -

No. 40 (PH 6)
Red Blood Cells absorb and store DARC-affinity chemokines
Hiroyuki Kayaba 1) , Toshihiro Shiratori 1) , Yumiko Yamauti 3) , Norihiro Saito 2) , Junichi Chihara 3) ,
Mihoko Kushibiki 1) , Hiroyuki Akimoto 1) , Shoji Tsutaya 1) , Keiya Kojima 1)
Clinical Laboratory, Hirosaki University Hospital 1) , Yokote Municipal Hospital 2) , Japan
Central Clinical Laboratory, Akita University Hospital 3) , Japan
Red Blood Cells (RBCs) express Duffy antigen receptor for chemokines (DARC) on the surface. DARC
is a decoy receptor with no signal transduction into RBC and binds to several Chemokines such as IL-8,
MCP-1, Eotaxin and RANTES. Biological function of DARC is thought to be a scavenger of
chemokines promoting chemokine concentration gradient to facilitate inflammatory cell migration in
the focus of inflammation. The aim of this study is to investigate how these chemokines scavenged by
RBC.
Blood (3.0 ml) was drawn from healthy volunteers aged 20 to 30 years and collected into EDTA tubes
containing K2EDTA (1.5 mg/ml). After centrifugation (430 G, 10 min), 200 µl of RBC was taken from
the bottom of the tube and washed three times with PBS. The expression of DARC and RANTES was
measured by flowcytometry (BD FACSCanto TM II). Intracellular content of RANTES was evaluated by
flowcytometry and ELISA (Quantikine, R&D). Other chemokines and cytokines including IL-8, MCP-1
and Eotaxin were measured by Bio-Plex Pro TM Assays, BIORAD).
DARC was expressed on RBC of all the samples examined. RANTES was recognized in RBCs, but not
on the surface. When compared the concentrations of IL-1β, TNF-α, VEGF, RANTES, Eotaxin MCP-1,
IL-8 in the washed RBC supernatant before and after hemolysis, RANTES, Eotaxin and MCP-1showed
marked increase after hemolysis. This finding showed that RBCs absorb chemokines which has high
affinity to DARC. RANTES was absorbed by RBCs rapidly. Intracellular RATNTES concentration
reached almost the maximum level within 15 minutes. DARC expression decreased in the presence of
RANTES.
RBC contains DARC-affinity chemokines. The role of RBCs in inflammation and immunological
diseases awaits to be explored.
- 93 -

No. 41 (PH 7)
Asian Thrombophilia: Dysfunction of the APC anticoagulation system.
Naotaka Hamasaki, Hiroaki Kuma, Hinako Hatae
Department of Clinical Chemistry, Faculty of Pharmaceutical Sciences, Nagasaki International
University, Japan
The anticoagulation system in a healthy individual is composed of mainly three systems, that is, (i)
tissue factor pathway inhibitor, (ii) antithrombin, and (iii) activated protein C (APC). The APC
anticoagulation system is unique in the sense that the anticoagulation activity is expressed only when
thrombin is formed by the coagulation system and the APC activity is regulated proportionately to the
coagulation activity.
The thrombophilia common among Caucasians is the genetic polymorphism of coagulation factor
V Leiden (R506Q). Factor V Leiden (R506Q) shows resistance to the APC anticoagulation system
(APC-resistance). Because of this, factor V Leiden (R506Q) carriers have relatively stronger coagulation
activity than the anticoagulation activity of APC, and is believed that this leads to excessive thrombus
formation and makes the person prone to thrombosis. Approximately 30% of Caucasian patients with
venous thrombosis are carriers of factor V Leiden (R506Q) [1]. Unlike Caucasians, thrombophilias
among Japanese and Chinese are the ones with dysfunction of the activated protein C (APC)
anticoagulant activity caused mainly by abnormal molecules of protein S or of protein C
(APC-dysfunction). Approximately 50% of Japanese and Chinese patients suffering from deep vein
thrombosis have reduced activities of the APC anticoagulant system [2]. In other words, the
APC-resistance in Caucasians and the APC-dysfunction in Japanese and Chinese are the major risk
factor for thrombosis. Whether the APC-dysfunction occurs in other Asian countries is an important
aspect of mapping thrombophilia among Asians, and international surveys would be needed to
determine it. Combined with our investigation and results of other studies, we will review the Asian
thrombophilia and indicate our strategy to overcome the “prone to thrombosis”.
1) Castoldi E, Rosing J: APC resistance: biological basis and acquired influences. J Thromb
Haemostat 2009; 8:445-453.
2) Hamasaki N: Unmasking Asian Thrombophilia: Is APC dysfunction the real culprit? J Thromb
Haemostat 2012; 10: 2016-2018.
- 94 -

No. 42 (PH 8)
Great impact of the brand-new Sysmex XN-Series automated hematology analyzer
on workflow efficiency and utility
Etsuko Hamada, Masato Maekawa
Department of Laboratory Medicine, Hamamatsu University School of Medicine, Hamamatsu, Japan
Introduction:
Recently, higher-quality testing environment has been increasingly required. To counteract these
demands, Sysmex has introduced the XN-Series automated hematology analyzer. Here we report our
evaluation results of the XN-Series basic performance and the integrated hematology testing system.
Method: (1) Basic performance: We evaluated precision, linearity, correlation with a previous analyzer
(XE-2100, Sysmex) and correlation with the manual method. (2) Usability: We evaluated the usability
of newly introduced functions of CNA-NET, which include automated smear preparation, selection of
specific sample and sample requiring manual count. Results: (1) Basic performance: Precision (CBC)
was 0.42-5.21 (CV%), and linearity was confirmed up to WBC 378×10 9 /L, RBC 7.81×10 12 /L, HGB 235
g/L, HCT 72.1%, and PLT 1,691×10 9 /L. The correlation coefficients (r) with the previous analyzer
were CBC 0.998–0.981, WBC classification ratio 0.995-0.917 and reticulocyte ratio 0.978. The
correlation coefficient of WBC classification ratio with the manual method was 0.612-0.987. In the
evaluation using samples with WBC ≤ 0.5×10 9 /L, WBC precision was 7.75-8.21 (CV%) for Whole
blood mode and 1.29-4.17 for Low WBC mode, and the good correlation with the manual method was
observed. In the evaluation using samples with PLT ≤ 50×10 9 /L, the correlation coefficient between
PLT-I (impedance) and PLT-F (fluorescence) was 0.973, and with the manual method was 0.975 for
PLT-I and 0.980 for PLT-F. (2) Usability: The number of analyzer operator to measure 700 samples/
day decreased from 1.5 to 1 person. Conclusion: Basic performance of the XN-Series was satisfactory.
Especially, precision and accuracy in the low number of WBC and PLT were excellent. Therefore it is
possible to report higher clinical value result with the automatic measurement method. Use of the
automated re-testing function and the CNA-NET made reporting time and processing time reduced. The
newly introduced integrated hematology testing system improved the quality of the clinical testing.
- 95 -

No. 43 (PH 9)
The comparison of the notations for quantitative evaluation of adhesive molecules’
expression on CD34 positive cells
Nobuo Masauzi 12 , Junji Tanaka 2 , Masaharu Kasai 3 , Masahiro Ogasawara 3 , Minami Doi 4 , Naoki
Kobayashi 3 , Yoshio Kiyama 3 , Minami Doi 4 , Sayaka Fukui 4 , Nao Fujimoto 4 , Misaki Yamada 4 ,
Keiko Miwa, 1 , Masanobu Kobayashi 5 ,Masahiro Imamura 3
1: Department of Medical Laboratory Science, Faculty of Health Sciences, Hokkaido University.
Sapporo, 2: Department of Hematology, Hokkaido University Graduate School of Medicine, Sapporo, 3:
Department of Hematology, Sapporo Hokuyu Hospital. Sapporo, 4: Department of Health Science,
School of Medicine, Hokkaido University, Sapporo, 5: Department of Nursing, Health Science
University of Hokkaido, Ishikari, Japan.
There are various quantitative notations for molecule expression on cells, such as the percentage of
positive expressing cells (PPC) and the mean fluorescence intensity (MFI). As to MFI, some evaluate
only MFI-PF among cells in positive fraction (PF), which express blighter fluorescence than that of
iso-type control anti-bodies, the others do MFI-PF&NF among cells in PF and negative fraction
(NF).We had published that the MFI-PF&NF of CD11a and CD11b on CD34 positive cells(CD34+) in
peripheral blood (PB) were inversely correlated to the yields of total collectedCD34+ before
administration of granulocyte-colony stimulating factor (G-CSF). Although some authors have
reported similar analysis, they represented the expressions of these adhesion molecules by PPC or
MFI-PF. Thus, the direct comparison of these results was impossible. We have analyzed the
significance of the change of PPC, MFI-PF and MFI-NF of CD11a and CD11b, and the correlations of
the yield of PB graft (YPBG). No significant changes ware indicated with 2-way ANOVA in PPC
neither by the number of days administrating G-CSF (Days), nor the YPBG, while there were
significant changes in MFI-PF&NF. The same test indicated not only significant change of MFI-PF of
CD11a by Days, but also that of MFI-NF of CD11a by YPBG. The significant correlation between the
PPC and the YPBG was not shown. MFI-PFs of CD11a and CD11b on day -1 indicated significant
inverse correlations with the YPBG. MFI-NFs of CD11a and CD11b indicated no significant
correlations with the YPBG The value of the lower fluorescence, which derived from non-specific
binding of anti-bodies, was commonly recognized as meaningless. The presenting results suggested that
the value of the lower fluorescence area had some important mean in a certain situation.
- 96 -

No. 44 (PH 10)
Relation between VKORC1 and CYP2C9 polymorphisms and warfarin dose
requirement in Japanese patients
Takeda Tomohiro, Mika Kataoka, Naoko Yamaguchi, Tizuko Kuramoto, Takao Uchiike ,Yasuyuki
Okamoto,
Central Clinical Laboratory, Nara Medical University Hospital, Japan
Among patients who take warfarin, the most commonly prescribed anticoagulant, there are great
differences in the dose required for sufficient effect. Recent reports have shown that the
polymorphisms of vitamin K epoxide reductase complex, subunit 1 gene (VKORC1) and cytochrome
P450, family 2, subfamily C, polypeptide 9 genes (CYP2C9) are closely associated with the dose
requirement of warfarin. In this study, we investigated factors related to the differences in the required
dose of warfarin, with special attention to the polymorphisms of VKORC1 and CYP2C9.
METHODS:
Relations between the dose of warfarin and PT-INR, age, body weight and the polymorphisms of
VKORC1 and CYP2C9 were studied in 86 stably anticoagulated patients with cardiovascular diseases.
The polymorphisms of VKORC1 -1639 G>A and CYP2CP 1075 A>C were determined by PCR-RFLP
method with the restriction enzymes NciI and KpnI, respectively.
RESULTS:
There was no significant relation between the dose of warfarin and PT-INR, age or body weight. The
AA and GA genotypes of VKORC1 -1639 were found in 69 and 17 patients, respectively. The mean
dose of warfarin required was significantly higher in the GA genotype (3.9 mg/day) than in the AA
genotype (2.4 mg/day, p

No. 45 (PH 11)
T-cell Acute Lymphoblastic Leukemia with Chromosomal Rearrangements
Involving the Immunoglobulin Heavy Chain Breakpoint at Band 14q32
Joonhong Park 1 , Myungshin Kim 1 , Jihyang Lim 1 , Yonggoo Kim 1 , Kyungja Han 1 , and Seok Lee 2
Department of Laboratory Medicine 1 , Division of Hematology 2 , Department of Internal Medicine, The
Catholic University of Korea, Seoul, Korea
T-cell acute lymphoblastic leukemia (T-ALL), a malignant proliferation of T-lymphoid blasts, represents
15% of acute lymphoblastic leukemia. In contrast with B-cell ALL, patients with T-ALL have different
chromosomal abnormalities and appear to have an unfavorable prognosis, leading to more intensive
treatment. Chromosomal rearrangements in T-ALL usually involve breakpoints in bands where the
T-cell receptor (TCR) genes are located; these are bands 14q11 (TCR-α and -δ genes), 7q32-36
(TCR-βgene), and 7p15 (TCR-γ gene). Other chromosomal changes, specific or not of T-ALL, have also
been observed. We report T-ALL with t(8;14)(q24.1;q32) similar to that seen in Burkett’s lymphoma.
Flow cytometric analysis of bone marrow revealed CD2, CD3, CD7, cytoplasmic CD3, CD34, HLA-DR,
CD13, and CD117. The phenotype was suggestive of T-ALL. RT-PCR screening for leukemia-related
fusion transcripts (HemaVision; Bio-Rad Laboratories, CA) was negative. Mitoses were obtained only
from spontaneously dividing cells in the absence of mitogens; 25 of the 30 metaphases analyzed were
chromosomally abnormal and had a der(14)t(8;14)(q24.1;q32). Disruption of the MYC gene which
resulted in a one fusion, one green, and one orange signal pattern was detected by MYC dual-color
break-apart probe (Abbott Molecular, IL) PCR-based B-cell and T-cell clonality (Ig and TCR gene
rearrangement) assays was studied by the BIOMED-2 multiplex Ig/TCRB PCR assay (InVivoScribe,
CA) but did not detected any clonal immunoglobulin and TCR rearrangements. Although abnormalities
involving 14q32 are characteristic of B cell disorders, they have also been described in T cell
malignancies, suggesting that genes transcribed in T cells and/or oncogenic sequences significant in T
cell neoplasia are present in 14q32.
- 98 -

No. 46 (PH 12)
Suspect of malaria in Sysmex XT series; case report of two cases
Iwan Joseph, Mansyur Arif, Hardjoeno
Department of Clinical Pathology, Faculty of Medicine Hasanuddin University, Makassar,
Indonesia
Malaria is an infectious disease caused by Plasmodium which is usually presenting fever. Microscopic
examination is gold standard test to diagnose malaria, but sometimes there might be error in laboratory
examination. Automated blood cell analyzer, for example, Sysmex XT2000i and XT1800i are
automatically tools for complete blood count including leukocytes differentiation and can be used to
suspect diagnosis of malaria early. Our case was a male patient of 44 years old who was admitted in
Wahidin Sudirohusodo Hospital of Makassar with chief complaint of fever. On the first day of
admission, stained blood film was negative for Plasmodium, but the Sysmex DIFF scattergram was
abnormal. Based on these findings, complete blood count and stained blood film were performed again
and we found trophozoites, schizonts and gametocytes of Plasmodium falciparum and Plasmodium
malariae. The second case a female patient of 37 years old had an abnormal DIFF scattergrams and we
found Plasmodium vivax in both thick and thin blood films. The results indicated that abnormal DIFF
scattergram of Sysmex analyzer could be used as an early clue or suspicion of Plasmodium infection,
especially if patient presented with chief complaint of fever. Abnormal areas of eosinophils and cell
ghosts as well as neutrophils in the DIFF scattergram may indicated the existence of Plasmodium
malaria.
Key words: Malaria, Sysmex XT, DIFF scattergram abnormal
- 99 -

No. 47 (PH 13)
Usefulness of platelet most frequent volume for platelet size evaluation in
thrombocytopenic patients
Setsuki ISONO 1) , Megumi TATSUTA 1) , Keiko NAGATA 1) , Keiko NUMATA 1) , Tosiaki
KOUJITANI 1) , Akiharu OKAMURA 1) , Masahumi TAKATA 2) , Mariko OKANO 3) , Humiko
HATANO 3) , Hiroyasu UEMURA 3) , Takeshi MORISAWA 3) , Masahiko YONETANI 3) , Mariko
TAKENOKUCHI 4) , Katsuyasu SAIGO 4)
1) Department of Laboratory Medicine, Kakogawa West City Hospital, Kakogawa, Hyogo. 2)
Department of Internal Medicine, Kakogawa West City Hospital, Kakogawa, Hyogo. 3) Department of
Pediatrics, Kakogawa West City Hospital, Kakogawa, Hyogo. 4) Faculty of Pharmacological Sciences,
Himeji Dokkyo University, Himeji, Hyogo, Japan
Introduction Platelet size information is helpful for differential diagnosis of the patients with
thrombocytopenia. Although mean platelet volume (MPV) is most frequently employed as platelet
size marker, occasionally MPV cannot be obtained mainly due to severe thrombocytopenia or widely
spread platelet size distribution. Then we have studied the usefulness of platelet most frequent volume
(P-MFV) as a surrogate platelet size marker. Materials and Methods Complete blood counts from
hematologically normal 100 adults, 44 children, 43 newborn infants( birth weight > 2500g) and 23
premature babies (birth weight < 2500g )were enrolled in this study by means of XE-2100 (Sysmex,
Kobe, Japan). They were outpatients or newborn babies in Kakogawa West City Hospital, between May,
2010 and May, 2012. Three patients with acute type immune thrombocytopenia, and one
Wiskott-Aldorich syndrome were also included. Results The correlation efficiency between P-MFV and
MPV was between 0.844 and 0.909 for all populations. P-MFV and MPV correlated very well in any
age group (p

No. 48 (PH 14)
Intravascular large B-cell lymphoma in Korea: Clinical and pathological findings
Hyun-Ki Kim 1 , Seongsoo Jang 1 , Chan-Jeoung Park 1 , Hyun-Sook Chi 1 , Jooryung Huh 2 and
Cheolwon Suh 3
Departments of Laboratory Medicine 1 , Pathology 2 and Internal Medicine 3 , University of Ulsan College
of Medicine and Asan Medical Center, Seoul, Korea
Background : Intravascular large B-cell lymphoma (IVLBCL) is a rare type of lymphoma characterized
by the selective growth of neoplastic cells within blood vessel lumina. In Western countries, the brain
and skin are the most common sites of disease. In Asian countries, mainly reported in Japan, the
IVLBCL manifest itself more frequently with fever, hepatosplenomegaly, hemophagocytosis, bone
marrow (BM) invasion, respiratory disturbance and disseminated intravascular coagulopathy. We aimed
to analyze characteristics of IVLBCL in Korea.
Methods : We retrospectively reviewed the records of 10 patients diagnosed as IVLBCL from January
1997 to December 2011 in Asan Medical Center. BM aspiration smears and biopsy sections were
obtained in 9 cases. H&E stain and Immunohistochemical stains (anti-CD3, anti-CD20, anti-CD79a and
anti-CD34) were performed on BM biopsies. And all cases were evaluated by Abdomen/Pelvis CT and
Chest CT. Whole body PET or brain imaging was taken in some cases.
Results : BM, lung and skin were frequently involved (3 cases, 4 cases, 2 cases, respectively).
Hemophagocytosis was observed in only one case. Total 4 out of 10 patients expired (40%). Two
patients with BM involvement suffered a relapse and expired (67%). Anti-CD20 and Anti-CD34
immunohistochemical stain helped with the diagnosis of BM involvement
Conclusion : Different clinical features from patients in Western countries or Japan. BM involvement
seems to be a strong unfavorable prognostic factor. Anti-CD34 ICH could be helpful for detection and
diagnosis of IVLBCL especially in cases with vague infiltration pattern on H&E stain.
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No. 49 (PH 15)
Evaluation of CellaVision DM96, an automated digital cell morphology
identification system, for the examination of body fluid cytospin slides
Hyun-Sook Chu, Young-Uk Cho, Sang Hyuk Park, Seongsoo Jang, Chan-Jeoung Park
Department of Laboratory Medicine, university of Ulsan, College of Medicine and Asan
Medical Center, Seoul, Korea
Background: Several automated digital imaging systems have been introduced in recent years and
showed excellent correlation with conventional manual and digital imaging differentials for the
evaluation of peripheral blood smears. The aim of this study was to evaluate one of the modern
automated digital imaging systems, the CellaVision DM96 (CellaVision AB, Sweden), for the
examination of body fluid cytospin slides.
Methods: We evaluated 101 body fluid cytospin slides from 91 patients. In the manual microscopic
differentials, 62 cases had malignant effusion or leptomeningeal seeding, of which 47 had metastatic
carcinoma and 15 had lymphoma or leukemia involvement. The remaining 39 cases showed benign
conditions. Specimen types included 40 peritoneal fluid, 26 pleural fluid, 25 cerebrospinal fluid (CSF),
5 bronchoalveolar lavage fluid, 3 dialysis fluid, and 2 pericardial fluid samples.
Results: Results from the comparisons showed good correlation for all cell classes with correlation
coefficients ranging from 0.643 to 0.998. Four CSF samples were not analyzed due to a very low
number of total leukocytes (< 2 /μL). Strong correlation was observed with neutrophils, lymphocytes,
macrophages and eosinophils with correlation coefficients greater than 0.8. However, correlation
coefficients for mesothelial cells and malignant cells were slightly lower, with the values of 0.698 and
0.643, respectively. More malignant cells were detected on manual microscopic examination than with
the digital imaging system, with a mean difference of 13.5%. Notably, cases with benign effusion or
CSF showed excellent correlation for all major cell classes, with correlation coefficients greater than 0.9.
Correlation was unaffected by the leukocyte count in the fluid.
Conclusions: Our evaluation of CellaVision DM96 revealed good correlation with manual microscopic
differentials for evaluation of body fluid cytospin slides. Thus, this system can be an effective option for
body fluid analysis in hematology laboratories needing an automated digital imaging system. In
particular, it can be used to rule out malignant effusion or leptomeningeal seeding as a first-line
screening tool and can help improve workflow and reduce turnaround time.
- 102 -

No. 50 (PH 16)
Calculation of Measurement Uncertainty for Complete Blood Cell Counts (CBC)
Atsushi Shirakami, Masako Koguchi, Tsutomu Kakuyama, Kanako Kiso, Takashi Kitagawa
Sysmex Corporation, Japan
Recently, due to penetration of ISO standards (ISO 15189 1) ) and globalization of health services,
medical laboratories are required to assure the reliability of testing results by showing its traceability
and measurement uncertainty. To achieve it, ISO 17511 2) requires IVD manufacturers to establish the
metrological traceability for the testing parameters to the end-users. For hematological testing
parameters, such as CBC, there is no certified reference material (CRM) except for Hb – fresh blood
sample is the only material that can transfer the data of the reference measurement procedure (RMP),
which made the standardization of CBC results more difficult than clinical chemistry. We established the
metrological traceability and the calculation methods of measurement uncertainty for the CBC 5
parameters, WBC, RBC, Hb, Ht and Plt, based on the GUM 3) approach. The major data-transfer process
in the manufacturer consists of the following three steps ;
Step 1. Value assignment of five fresh blood samples with RMP
Step 2. Calibration of standard analyzers with value-assigned fresh blood samples
Step 3. Value assignment of product calibrator
For each step, we evaluated the major uncertainty sources and calculated the measurement uncertainty
by ANOVA and uncertainty budget. Then we assigned the values of the product calibrator with
expanded uncertainty (coverage factor; k=2).So medical laboratory can assure the traceability and
measurement uncertainty by calibrating the hematology analyzer with the product calibrator. However,
as for utility of uncertainty in the routine laboratory test results for clinician, more discussion is
necessary.
1)
ISO 15189, Medical laboratories – Particular requirements for quality and competence
2)
ISO 17511, In vitro diagnostic medical devices - Measurement of quantities in biological
samples - Metrological traceability of values assigned to calibrators and control materials
3)
GUM : Guide to the Expression of Uncertainty in Measurement
- 103 -

No. 51 (PH 17)
Serum Thymidine Kinase 1 as a Potential Marker for Aggressive Behavior in
Patients with B-cell Lymphoma
Heyjin Kim, Jin Kyung Lee, Young Jun Hong, Seok-Il Hong, and Yoon Hwan Chang
Department of Laboratory Medicine, Korea Cancer Center Hospital, Seoul, Korea
Aim: B-cell lymphoma comprises a heterogeneous group of lymphoproliferative malignancies with
various clinical and biological features. The cell cycle-dependent enzyme, thymidine kinase 1 (TK1) has
been known as one of the markers of cancer cell proliferation and had been reported as a prognostic
factor in various solid tumors and CLL. The aim of this study is to evaluate the usefulness of TK1 as a
biomarker for aggressive behavior in patients with B-cell lymphoma.
Methods: A total of 52 patients with previously untreated B-cell lymphoma (25 women, 27 men; age,
14-86 years; median age, 62.5 years) and 43 healthy control subjects were enrolled in this study. TK1
level in serum were measured by chemiluminescence immunoassay (LIAISON ® , DiaSorin, USA).
Results: The 95 percentile reference range for serum TK1 in healthy controls was 4.5-18.6 U/L. There
was a clear difference in mean TK level was shown between patients and healthy controls (46.8 ± 79.0
vs. 9.9 ± 3.9 U/L, P = 0.003). The mean TK1 level of patients with bone marrow involvement was
significantly elevated compared with that of patients without bone marrow involvement (128.8 +± 144.1
vs. 29.6 ± 43.3 U/L, P=0.002). The mean TK1 level in the clinical stage III and IV group was
significantly higher than that of the clinical stage I and II (85.1 ± 109.8 vs. 20.8 ± 28.9 U/L, P=0.016)
group. The increased TK1 level at the time of diagnosis was correlated with the advanced clinical stage
(P=0.000), international prognostic index score (P = 0.001), B symptoms (P = 0.035), hemoglobin level
below 12.0 g/dL (P=0.004), and lactate dehydrogenase level above 480 U/L (P=0.000).
Conclusions: These findings indicate an association between serum TK1 level and aggressive behavior
of B-cell lymphoma. Therefore, the TK1 level could serve as a valuable prognostic marker in patients
with B-cell lymphoma.
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No. 52 (PM 1)
High seroprevalence of human herpes virus type 8 in patients with lung carcinoma
Cheng-Chuan Su 1,2,3 , Chun-Liang Lai 4 , Ming Nan Lin 5
1
Institute of Medical Sciences, and Departments of Laboratory Medicine and Pathology, School of
Medicine, Tzu Chi University, Hualien, Taiwan; Departments of 2 Clinical Pathology, 3 Anatomic
Pathology, 4 Chest Medicine, and 5 Family Medicine, Buddhist Dalin Tzu Chi General Hospital, Chiayi,
Taiwan
Background: Human herpes virus type 8 (HHV-8) DNA is found consistently in all types of Kaposi’s
sarcoma (KS), which is sometimes seen in human immunodeficiency virus (HIV) non-infected patients
with immunologic abnormalities. Lung carcinoma is one of the most common malignancies developing
in immunocompromised patients. However, the prevalence of HHV-8 infection in lung carcinoma
patients is unclear.
Methods: Blood samples were collected from 109 lung carcinoma patients with malignant pleural
effusion and 109 age-matched healthy controls and analyzed for lymphocyte and monocyte counts, and
presence of HHV-8 antibody and DNA. All study subjects were negative for anti-HIV antibodies.
Results: Lung carcinoma patients had significantly lower mean lymphocyte counts and significantly
higher monocyte counts than the healthy controls (P < 0.001). Three patients with lymphopenia and
stage IV tumor were positive for HHV-8 DNA, one of them was negative for HHV-8 antibody. HHV-8
positivity was significantly higher in patients (42.2%), particularly in male patients (50.8%), than in
healthy controls (24.8%) (P = 0.006 and < 0.001, respectively). HHV-8 positivity was significantly
greater in male patients than in female patients (29.5%) (P = 0.028). HHV-8 antibody titers in patients
also significantly exceeded those in healthy controls (P = 0.004). All subjects positive for HHV-8 were
not associated with clinical manifestations of HHV-8 infection.
Conclusions: HHV-8 seroprevalence was significantly greater in lung carcinoma patients than in
healthy controls, and associated with gender.
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No. 53 (PM 2)
HIV-1 RNA viral load in seminal fluid: quantification with NASBA
Lyana Setiawan, Agus Susanto Kosasih, Samsuridjal Djauzi, Haridana Indah Mahdi,
Runingsih
Dharmais Cancer Hospital, Indonesia
As the prevalence of HIV infection increases, the problem of discordant couples with HIV arises.
Although the number is yet unknown, discordant couple presented problem of possible infection to the
mothers and unborn babies. In this study, we assessed the utilization of nucleic acid sequence based
amplification (NASBA) to quantify HIV-1 RNA in seminal fluid, and reported the HIV-1 RNA viral
load in the seminal fluid of patients with discordant couple from the outpatient clinic of Dharmais
Cancer Hospital during July 2010 – February 2012, who have already been successfully treated with
antiretrovirals (ARV).
To determine the usefulness of NASBA, RNA from 2 unprocessed seminal fluid specimens were
extracted using the MiniMag®, amplified by NASBA, and the result quantified. To verify the usefulness
of sperm washing, we processed specimen with high HIV-1 RNA with gradient centrifugation to wash
the sperm, and quantified the viral load in the resulting seminal plasma from first centrifugation, and the
supernatant and sperm from the second washing. After optimization, we evaluated the result of 43
patients treated with ARV who were referred for HIV-RNA in seminal fluid test.
HIV-1 RNA was detected in the unprocessed seminal fluid (Pt#1 19,000 IU/ml; pt#2 740 IU/ml), and
in the seminal fluid from the first centrifugation. Supernatant and sperm from the second washing
showed HIV-RNA below detection limit. The result proved that boom extraction and NASBA principle
can be used for quantification of HIV-RNA in seminal fluid, and that sperm washing was effective to
minimize the risk of infection in assisted reproduction, but this needed to be verified with more samples.
All 43 patients showed HIV-RNA below detection limit (

No. 54 (PM 3)
Epidemiological and microbiological feature of meningococcal infection in
Ulaanbaatar city
B. Uyanga, MD 1 , G. Oyungerel, MD 2
1 SOS Medica Hospital, Ulaanbaatar, Mongolia
2 Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
Objectives: Detection of carrier state of meningococcal infection and study of their distribution in
different area of Ulaanbaatar city for the period of January – April 2011.
Materials and Methods: 4720 throat swabs were collected and inoculated on Thayer – Martin agar
medium, as used for primarily isolating Neisseria meningitides and as the medium that inhibits the
growth of the most other microorganisms. Biochemical properties of the isolated strains characterized
on medias, enriched with different carbohydrates such as glucose, maltose, saccharose and lactose.
Standardized methods have been used for testing of oxidase activity and serological typing.
Results: For the period of 2005 – 2010 registered several cases of spreads of meningococcal meningitis
in the city, preceded by increase of “healthy” carriers of meningococci, who considered as the source of
spreads to the surrounding people. 514 samples give growth of the N. meningitides, where serogroup B
detected as most prevalent type (69.74%), followed by serogroup C (10.3%) and serogroup A as the
lowest type (0.19%), with irregularity in distribution in different parts of the city.
Highest positivity of carrier states were revealed geographically in Sukhbaatar and Bayan – Gol districts
as taken together (70% of the total growth), and peak of the positivity of months of January and
February followed by marked decline in the month of April. Age group of positivity falls between 0 and
7 years (65%), as expected and most of them were from kinder garden and only 27 kids were from home
staying places.
Conclusion: 1. Serogroup B is the main type (~70%) of the meningococcal strain in Ulaanbaatar city
and serogroup A is the least one (0.19%), with no registered serotype of D and Y. 2. Epidemiologically
important carrier states were detected mostly in children visiting communal institutions like kinder
gardens and schools (405 cases) and were characterized by seasonal and geographical differences in the
city.
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No.55 (PM 4)
The inhibitory effect of serum HLA class I antigens on EBV-specific CD8+CTL in
vitro
N.Erdenesuvd 1,3 , B.Gansuvd 1,2 , B.Munkhbat 1,2 , M.Hagihara 2 , T.Hotta 2 , N.Munkhtuvshin 1 ,
1
Central Scientific Research laboratory, Nat’l Institute of Medicine, Mongolia
2
Department of Transplantation Immunology, Tokai University, Japan
3
Dept. Laboratory Medicine, General Hospital, Uvurkhangai, Mongolia
Objectives: The level of sHLA class I molecules in human sera is markedly increased after BM and
organ transplantation, viral infections, autoimmune disorders and cancer. Their biological and clinical
significance has not been fully studied. In the present study the effects of soluble HLA (sHLA) class I
molecules against EBV-specific CTL were examined.
Materials and Methods: Two different sources of sHLA class I, either bioengineered spliced form of
HLA-B7 (sB7) or natural production from EBV-transformed B cells (natural sHLA), were added during
the induction of CTL or incubated with MHC-restricted CD8+ CTL, which were selected by
immunobeads just before testing for their cytotoxic activity.
Results: Both sB7 and natural sHLA class I blocked the generation of CD+ CTL and also inhibited the
cytotoxic activity of established CTL in a dosedependent manner. In both ways, natural sHLA class I
was effective in 10-fold lover concentrations compared with sB7. The inhibitory effect did not require a
shaping of the HLA allotypes between sHLA and the CTL. CTL, after being treated with sHLA,
underwent apoptosis, which was considered here as the main mechanism. Generally, sHLA class I
consists of three different molecular forms: a shedding form with a heavy chain of 44 kD, a splicing
form with a heavy chain of 35 kD and 37 kD. All these molecules are associated with β2-microglobulin.
Also it was reported that membrane-derived, naturally occurring sHLA class I forms multimers through
the ydrophobic
transmembrane tail, whereas cytosol-derived sHLA class I cannot form aggregates. In our studies, the
naturally sHLA class I consisted of all three different molecular forms of HLA-B7.
Conclusion: The naturally occurring sHLA class I induces apoptosis more markedly than sB7. supports
the same suggestion that the membrane -derived naturally occurring sHLA class I could cross-link with
TCR/CD3 complexes via its multimeric forms, and it probably leads to more effective apoptosis of the
CTL. The difference in the biochemical nature of sHLA class I molecules might be the reason why the
inhibitory effect of naturally occurring sHLA class I as stronger than that of sB7.
- 108 -

No. 56 (PM 5)
Tuberculosis screening by a T cell interferon-γ release assay in students of medical
school and international students in Gunma University
Rumi Watanabe 1) , Takao Kimura 2) , Yutaka Tokue 3) , Takayuki Ogiwara 3) , Makoto Nara 3) , Yoshino
Kobayashi 1) , Toshiya Inoue 1) , Hiroyuki Sumino 1) , Tadashi Morimura 1) , Osamu Araki 1) , Katsuhiko
Tsunekawa 1) , Tomoyuki Aoki 1) , Toshiko Obuchi 3) , Yukie Yomoda 1) , Kihachi Ohshima 2) , Masami
1, 2, 3)
Murakami
1) Clinical Laboratory Center, Gunma University Hospital, Maebashi, Japan. 2) Department of Clinical
Laboratory Medicine, Gunma University Graduate School of Medicine, Maebashi, Japan, 3)Infection
Control and Prevention Center, Gunma University Hospital, Maebashi, Japan
Screening and targeted testing for tuberculosis (TB) is a key strategy for controlling and preventing
infection on university campus and teaching hospital. The Mycobacterium tuberculosis antigen-specific
interferon-γ release assays (IGRAs) are used to detect latent tuberculosis infection. In Japan, it is
desirable to obtain basal data of IGRAs in medical school students and international students. To
evaluate tuberculosis risk in university campus and medical school at course entry, retrospective study
was performed. One thousand two hundred seventy-one students (887 medical students and 384
international students in Gunma University) underwent QunatiFERON-TB Gold test (QFT-TB) or
QunatiFERON-TB In-Tube test (QFT-GIT) at course entry. Eight of 887 medical students (0.9%) were
positive for QFT-TB or QFT-GIT, and none was diagnosed to have active tuberculosis among positive
students. Thirty of 384 international students (7.8%) were positive for QFT-GIT, and 2 students of 30
QFT-GIT positive students (6.7%) were diagnosed active tuberculosis during follow up. Positive ratio of
QFT-GIT in international students of Gunma University was significantly higher than that of medical
students. We propose tuberculosis screening with QFT-GIT test as a standard approach for medical
students and international students in Japan.
- 109 -

No. 57 (PM 6)
Differences of Vancomycin MICs for MRSA Isolates Based upon the study
Susceptibility Test Method Used
K Watanabe, M Mikami, Y Saya, C Tanaka, F Omata, K Furukawa, K Takeda
St. Luke’s International Hospital, Japan
Background: Recent studies reported increasing trend of vancomycin (VCM) treatment failure in
methicillin-resistant Staphylococcus aureus (MRSA) blood stream infections. Differences in VCM MIC
values among different tests are also reported. The aim of this study was to evaluate recent change in the
prevalence of MRSA with high VCM MIC(=2.0) and to compare the results of the three methods such
as E-test®, VTEK2® and Broth microdilution (BMD). Methods: E-test® ,VITEK2® and BMD were
applied for measuring VCM MIC in consecutive 59 MRSA strains isolated from blood cultures of 59
patients between 2007 and 2012. The tests results were categorized to binary variables using cut-off
value of 2.00μg/ml to calculate the kappa statistic. The annual change of prevalence of VCM resistant
MRSA was tested by Fisher’s exact test.
Results: MRSA were isolated from blood cultures in 28 cases in 2008,12 cases in 2009, 9 cases in 2010,
and 10 cases in 2011. The prevalence of strain with VCM MIC 2.0μg/ml using BMD method was 14%
in 2008, 28.6% in 2009, 0% in 2010, and 10% in 2011. There was no significant annual change in the
prevalence of VCM resistant MRSA (P=0.23 by Fisher’s exact test).The proportion of strain with MIC
2.00μg/ml of BMD, VITEK2® and E-test® method was 15.2%, 0.5% and 59%, respectively. The kappa
statistic of three tests was 0.12. Conclusion: There is no significant change in the prevalence of MRSA
with high VCM MIC (=2.0) measured by BMD method from 2008 to 2011. Comparing to BMD method,
VITEK2® method tends to miss MRSA with high VCM MIC (=2.0). On the other hand, E-test®
method tends to overdiagnose MRSA with high VCM MIC (=2.0). The differences between tests’
characteristics should be taken into account for clinical implication
- 110 -

No. 58 (PM 7)
Mycobacterium Kyorinense Infection: Clinical Features and Antimicrobial
Susceptibility
Hiroaki Ohnishi 1 , Shota Yonetani 1 , Satsuki Matsushima 1 , Kouki Ohtsuka 1 , Tomonori
Kishino 1 , Hiroo Wada 2 , Hajime Goto 2 , Takashi Watanabe 1
Departments of 1 Laboratory Medicine and 2 Respiratory Medicine, Kyorin University
School of Medicine, Japan
Mycobacterium kyorinense is a nonpigmented, slowly growing mycobacterium that was initially
isolated in 2007. Biochemical tests and genetic analyses showed that M. kyorinense is most closely
related to M. celatum and M. branderi. We here describe 7 newly identified cases with 4 previously
reported cases, in which infection potentially was caused by M. kyorinense. In reviewing these 11 cases
(10 Japanese and 1 Brazilian), 9 presented with respiratory infections, 1 with lymphadenitis, and 1 with
arthritis. Seven patients were treated by first-line tuberculosis drugs, mainly consisting of rifampin,
isoniazid, and ethambutol, but these therapies were ineffective in all cases. Six cases were treated with a
combination of antibiotics including macrolides and fluoroquinolones as a first- or second-line
chemotherapy, and infection was subdued without recurrence in 5 cases. In contrast, 4 pneumonia
patients who did not receive sufficient therapy with the latter regimen eventually died of infection. In
concordance with the clinical antimicrobial susceptibility, most strains exhibited relatively high MICs
for rifampin, ethambutol, and isoniazid, and relatively low MICs for macrolides, aminoglycosides, and
quinolones. Notably, remarkably high MICs (>32 �g/ml) w
Direct sequencing of the 16S rRNA gene revealed that all available M. kyorinense isolates were
identical within this gene, except for the Brazilian strain which showed slight difference with other
Japanese isolates. Direct sequencing of the entire rpoB gene demonstrated that all strains had identical
sequences, with Ser531 in the M. tuberculosis RpoB protein replaced by an Asp in M. kyorinense.
Notably, Ser531 is the most frequent location of substitutions in rifampin-resistant strains of M.
tuberculosis. These data suggest that M. kyorinense belongs to non-tuberculous mycobacteria that have
pathogenicity for humans with substantial clinical significance, and that M. kyorinense is inherently
resistant to rifampin due to the structural features of its RpoB protein.
- 111 -

No. 59 (PM 8)
The first case of Lecythophora hoffmanni peritonitis in Korea
Hunsuk Suh 1 MD, PhD, Jonghee Shin 2 MD,PhD
1. Laboratory medicine, Daegu Catholic University Hospital 2. Laboratory medicine, Jeonnam
University Hospital, Korea
The Lecythophora species are very rare pathogen to human reported as subcutaneous abscess, corneal
infection, endophthalmitis, sinusitis, peritonitis, and endocarditis, and cannot found the clinical case
especially caused by Lecythophora hoffmannii in Korea. So we introduce the first case of L. hoffmanni
peritonitis infection in Korea.
A 79-year-old female had a medical history of end stage of renal disease (ESRD) with continuous
ambulatory peritoneal dialysis (CAPD) was hospitalized for abdominal pain with diarrhea and anorexia
at June 2012. Initial WBC counts were 11.9 x 10 3 /uL, Hb was 11.1 g/dL, and platelet were 643 x 10 3 /uL.
ESR were 104 mm/hr and BUN and Cr were 61.8 and 6.6 mg/dL, respectively. The abdominal CT
showed sclerosing encapsulating peritonitis, so she treated by experience antibiotics. Cultures was done
with 7 peritoneal fluid samples from June 13 to July 16, same fungus was growth all samples. The rate
of growth is average 4 days at 30C. The colony morphology was smooth, moist to slimy yeast like, with
fuzzy. It shows yellow-orange to salmon pink colors. Reverse is yellow to orange. In microscopic, yeast
like conidia were significantly noted and a few hyaline, septated hyphae were seen. It was not identified
used by yeast card of VITEK II system (Biomerieux, French). So the 26S rRNA sequencing was done
and identified as Lecythophora hoffmannii.
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No. 60 (PM 9)
Sensitive, one, two-drug resistance, and MDRP Pseudomonas aeruginosa integron
carrying rates
Daisuke Tanabe
Bunkyo Gakuin University graduate school, Japan
Pseudomonas aeruginosa is pathogenic bacteria isolated high frequency to cause opportunistic infection.
Since the introduction of antibiotics in the treatment of P. aeruginosa infection, antibiotic resistance
has spread quickly among bacteria. Such explosive dissemination is a consequence of the mobility of
the antibiotic resistance encoding genes, which are usually found in integrons. That resistance
mechanism called integron structures is associated with the insertion of resistance genes into cassette
region. Although, the analyses of integron structures of MDRP has been reported, the analyses of
sensitive, one, and two-drug resistant isolates has less reported. The purpose of this study is to analyze
integron structures of P. aeruginosa and to investigate carrying rates. P. aeruginosa was used 96
clinical isolates from hospitals located in the Kanto area. Susceptibilities which used 10 antimicrobial
agents was determined by MIC testing in reference to Clinical and Laboratory Standards Institute
(CLSI). SMA disk was used to screen for Metaro β-lactamase producing (MBL) isolates. PCR
analyses for the detection of MBL genes were carried out for all strains for which the screening test
using SMA disks gave positive results. Furthermore, class 1,2,3 integrons were performed all isolates
according to the method reported by Senda. From the Susceptibility testing result, P. aeruginosa was
separated into 4 classes (sensitive, one, two-drug resistant, and MDRP) according to MDRP criteria of
CLSI (2010) (IPM 16µg/ml, CPFX 4µg/ml, and AMK 32µg/ml). There were Forty-six, twenty, sixteen,
fourteen, respectively. Thirty isolates were carried class 1 integron; two of 46 sensitive isolates, 7of 20
one-drug resistance, 8 of 16 two-drug resistance, and 13 of 14 MDRP. In this present study, the parts of
sensitive, one , and two-drug resistance isolates carried class 1 integrons. It is considered that
non-MDRP may be able to change MDRP according to insert resistance genes into integrons.
- 113 -

No. 61 (PM 10)
Application of MALDI-TOF MS-based strain typing for characterization of
epidemiological relationships among bacterial strains
Megumi Oho 1) , Zenzo Nagasawa 1) , Koji Kusaba 1) , Takanori Higashitani 1) , Shoitiro Ohta 1) ,
Eisaburo Sueoka 1) , Hiroshi Miyamoto 2)
Department of Laboratory Medicine, Saga University Hospital 1) ,
Division of Microbiology, Department of Pathology and Microbiology,
Faculty of Medicine, Saga University 2) , Japan
【Purpose】MALDI-TOF MS-based strain typing for microorganisms is a fast and inexpensive
technology for identification of bacteria. We introduced MALDI Biotyper Ⓡ (Bruker Daltonics Inc,) as a
clinical diagnostic system for microbial infection since April this year. The system needs only 10
minutes from isolation of the target colonies and considerably accurate compared with genomic analysis
using 16S rRNA. We applied this system to characterize the epidemiological relationships among
bacterial strains.
【Samples and Methods】24 of MRSA and 48 of Pseudomonas aeruginosa strains, those genomic
pattern determined by pulsed-field gel electrophoresis (PFGE), were analyzed by MALDI Biotyper, and
the characteristic amino acid profiles were assessed by ClinProTools. For extraction of the proteins from
bacteria, an ethanol-formic acid extraction method was used, and the assays were conducted by
duplicate to obtain confirmative spectra. The spectra were analyzed by using ClinProTools (Bruker
Daltonics Inc.). ClinProTools is designed to facilitate the processing and comparison of multiple spectra
from each bacterium by automatically normalizing, baseline subtracting, peak defining, and
recalibrating.
【Results】We compared the results obtained from MALDI-TOF MS combined with ClinProTools and
those from PFGE. The epidemiological relationships examined by MALDI-TOF MS and PFGE among
MRSA and Pseudomonas aeruginosa strains were almost identical. The results were validated with
other molecular epidemiological examinations.
【Conclusion】The epidemiological relationships examined by MALDI-TOF MS showed comparable
data with PFGE. Since MALDI-TOF MS-based strain typing for microorganisms is a fast and
inexpensive technology for identification of bacteria, the technique combined with ClinProTools is
thought to be a useful tool for molecular epidemiological relationships among bacterial strains.
- 114 -

No. 62 (PM 11)
Simultaneous inhibition of NF-κB and caspase-1 by a cell-permeable compound
DHMEQ leads to potent suppression of IL-1β
Sayaka Ito, Misako Iwata, Tetsuo Kubota
Graduate School of Health Care Sciences, Tokyo Medical and Dental University, Japan
Objective: A rare autoinflammatory disorder, cryopyrin-associated periodic syndromes (CAPS), is
caused by uncontrolled secretion of IL-1β leading to recurrent inflammatory attacks. To develop a novel
therapeutic strategy for CAPS, we studied an effect of a cell-permeable compound (-)-dehydroxy methyl
epoxy quinomicin (DHMEQ) in an in vitro assay system for IL-1β production.
Methods: Peripheral blood mononuclear cells (PBMCs) from patients with CAPS and healthy
volunteers were primed for 3h with LPS, followed by stimulation with ATP, and secreted IL-1β was
measured by ELISA. DHMEQ (provided by Dr. Umezawa, Aichi Medical University) was added at
different time points to examine the effect on expression of IL-1β and activation of caspase-1. Activated
caspase-1 was detected by fluorescent staining using FAM-Tyr-Val-Ala-Asp-FMK. Caspase-1 and IL-1β
in the cell lysates and culture supernatants were detected by western blotting. Recombinant caspase-1
enzyme activity was measured by colorimetry using Ac-Tyr-Val-Ala-Asp-pNA.
Results: Normal PBMCs secreted IL-1β only after ATP stimulation following LPS priming. In contrast,
CAPS patients’ PBMCs secreted IL-1β even without ATP stimulation. In both cases DHMEQ effectively
suppressed IL-1β secretion when it was added to the culture before LPS priming. Western blotting
revealed suppression of proIL-1β expression during the LPS priming, which was thought to be one of
the mechanisms of the effect. In addition, when we added DHMEQ after expression of proIL-1β, it
significantly inhibited IL-1βsecretion. Results of western blotting and cytostaining suggested that
DHMEQ inhibited activation of caspase-1. In a cell-free system, DHMEQ inhibited the enzyme activity
of recombinant caspase-1 in a dose dependent manner.
Conclusion: DHMEQ is a derivative of an antibiotic epoxy quinomicin C, designed for the NF-κB
inhibitory activity. In addition, we found herein the inhibitory effect of DHMEQ on caspase-1 resulting
in a potent suppression of IL-1β secretion without significant cytotoxicity. DHMEQ might be a good
candidate drug for clinical application in therapy of CAPS.
- 115 -

No. 63 (PM 12)
Comparison of a novel real-time PCR system in the quantification of HIV-1 RNA
Demak Lumban Tobing, Sri Hartini, Theresia Koeshandini, Runingsih
Dharmais Cancer Hospital Clinical Pathology Laboratory, Indonesia
HIV-AIDS is a major healthcare problem in Indonesia, due to increasing prevalence, lifelong therapy,
and high cost for monitoring. Monitoring of therapy requires HIV-RNA quantification, which is
expensive and requires special equipment, dedicated room and skilled man power. A novel real-time
PCR system (Existation) had been developed, which enabled real-time measurement of HIV-1 RNA
in small to medium laboratory. We evaluated the performance of this system as compared to the
reverse-transcriptase PCR (Cobas Taqman) and nucleic acid-based sequence amplification/ NASBA
(Nuclisense). Fifty nine plasma samples were run on both Cobas Taqman and Existation, and 30
plasma samples run on Nuclisense and Existation. The result of HIV-1 RNA viral load were
analyzed descriptively. Of the 59 samples, 38 were not detected with Cobas Taqman (lower detection
limit 33 IU/ml), and 3 of the 21 positive samples were below 400 IU/ml; 33 samples showed negative
result from both platforms; 17 positive samples showed good agreement with the result of Existation
consistently lower than Cobas Taqman (-1.22 log 10

No. 64 (PM 13)
Measurement of Procalcitonin (PCT) and C-Reactive Protein (CRP) in Patients with
ESBL-negative Bacteremia
Soohun Yoo 1 , Seri Jeong 1 , Yongjung Park 1 , Nam Su Ku 2 , Dongeun Yong 1 and Hyon-Suk Kim 1*
1 Department of Laboratory Medicine and 2 Internal Medicine, Severance Hospital, Yonsei University
College of Medicine, Seoul, South Korea
Aim: We measured procalcitonin (PCT) and C-reactive protein (CRP) in patients with extended
spectrum beta lactamase (ESBL)-negative bacteremia and analyzed the results according to clinical
status of the patients. Materials and Methods: A total of 100 patients who were ESBL-negative
bacteremia were analyzed. Among measured database, data from first and last tests of PCT (PCT1,
PCT2) and CRP (CRP1, CRP2) during hospital stay were evaluated. PCT levels were measured by the
VIDAS ® BRAHMS PCT assay, and CRP concentrations were determined by a turbidimetric assay using
CA-400 analyzer. Results: The levels PCT1 of patients with community acquired infection (n=65) were
more significantly higher than those with nosocomial infection (n=35) (12.6 ng/mL vs. 1.5 ng/mL,
p=0.0001). Clinically, PCT1 levels were significantly higher in patients with central line insertion
(p=0.0089), pneumonia (p=0.0233) and intensive care unit admission (p=0.0258) than in patients
without these conditions. CRP2 levels were significantly higher in patients with intubation (p=0.0064),
leukocytosis at the admission (p=0.0081) and urinary tract infection (UTI) (p=0.0198). The higher
levels of PCT2 were shown in patients with mortality (p=0.0005) and UTI (p=0.0198). Similarly,
CRPs2 were also relevant with mortality (p

No. 65 (PM 14)
Significant Increase of Sensitivity on Rapid Influenza Antigen Assays Using Silver
Amplification Immunochromatography
Satoshi KIMURA 1) , Hideyoshi OHTO 2) , Hayato YAMAGUCHI 1) , Seiji FUKUOKA 1) , Emiko
NAKAMA 1) , Akiko TERADA 3) , and Akira UMEDA 2)
1) Department of Laboratory Medicine and Central Clinical Laboratory, Showa
University Northern Yokohama Hospital, Yokohama 224-8503, Japan
2) Department of Pediatrics, Showa University Northern Yokohama Hospital
3) R & D department, FUJIFILM Medical Co., Ltd, Tokyo, 106-0031, Japan
Introduction: Influenza virus has been recognized as one of the most pandemic infectious disease agent.
Rapid diagnosis of influenza is commonly performed with immunochromatography method (IC) using
specimens taken from upper respiratory tract. Although IC is easy and relatively cheap, its sensitivity
is not perfect especially in early stage of disease because of low concentration of virus antigens.
Applying a newly developed “silver amplification” principle, we generated a new assay method to
increase sensitivity. The aim of this study is if our method has higher sensitivity and specificity than
conventional IC assays.
Materials and Methods: One hundred and twenty cases of influenza-like symptoms; fever, rhinorrhea,
cough, and/or general fatigue who visited pediatrics department of our hospital from November 2011 to
April 2012 were entitled. Cotton swab specimens were applied to Fuji Drychem IMMUNO AG1 TM
(FUJIFILM Medical, Tokyo, Japan). Simultaneously, a conventional IC method was performed with
Quick Chaser FLU A/B TM by Mizuho Medy (Tosu, Japan). A real time PCR with TaqMan probe was
also done for gold standard. This study was authorized by local ethic committee and written informed
consent was obtained from all the cases.
Results: With silver amplification method, 23, 15 cases were influenza A, B positive, respectively. On
the other hand, 23, 8 cases were A, B, positive with conventional assays. All the cases, which showed
positive in new, negative in conventional influenza B assays, PCR results were positive. Their virus
concentration ranged 10 5 to 10 9 copies/ml.
Discussion: Specificity and sensitivity for influenza A was not different between the two assays.
However, the silver amplification method showed higher sensitivity for influenza B. Because the new
method applies photo-developing principle, its sensitivity was reported to increase 8 to 16 times,
theoretically. This analyzer is small (18x20x11 cm) and light (1.8kg), it is suitable for bedside testing.
Since number of cases is limited, more data are required to confirm the result.
Conclusion: Our novel assay method using silver amplification has high potentiality to increase
sensitivity of rapid bedside testing especially for influenza B.
- 118 -

No. 66 (PF 1)
Comparison of four portable apnea recording devices in daytime screening session
for severe sleep apnea patients screening
Chikage Torisawa, Chie Watanabe, Takeshi Tomihara, Chisato Shimazu, Taiji Furukawa
Department of Laboratory Medicine, Teikyo University School of Medicine, Japan
Evaluation for sleep apnea syndrome (SAS) in patients with cardiovascular problems is recommended in
several existing guidelines. Although overnight polysomnography (PSG) is the golden standard for the
evaluation, access to the examination is limited because it requires special institution and trained
technicians. Therefore, many portable recording devices with SpO2 monitoring only and those with
SpO2 and airflow monitoring capability are developed for SAS screening. We constructed a daytime
SAS screening session using portable devices to find out SAS patients during routine clinic visit of
patients with cardiovascular problems. The subjects were asked to remain quiet and in a supine position
in a dark room during the session for about 30 min. The present study evaluated the usefulness of four
portable devices in detecting severe SAS cases through the screening session. The four devices are a
thermister airflow-sensor type device (FM-500, Fukuda-denshi, Japan), two pressure airflow-sensor type
devices (LS-300, Fukuda-lifetech; Morheus, Teijin, Japan), and a sheet-type device that detects chest
wall movement (SD-101, Suzuken, Japan). Two hundred and ten patients underwent PSG after the
screening session using one of the portable devices. We compared the respiratory disturbance index
(RDI) measured in the sessions and apnea-hypopnea index (AHI) measured by standard PSG performed
afterward. The RDI by all devices and AHI correlated significantly. ROC analysis revealed that the
thermister type device and sheet type devices had larger area under the curve (0.78 and 0.77,
respectively) than two pressure sensor type devices (0.71 and 0.73) in detecting severe SAS patients
(AHI in PSG≧ 30). The difference might be related to condition of airflow recording under nasal
obstruction, as mouth breathing could not be monitored by the pressure type sensor in 8 cases. The
results suggested that portable recording devices usable under nasal obstruction would be suitable for
SAS screening session.
- 119 -

No. 67 (PF 2)
The relationship between arterial wall elasticity by the phased tracking method and
vascular manifestations in type 2 diabetes mellitus patients
Michiaki Miyamoto 1) , Kazuhiko Kotani 1) , Hideyuki Hasegawa 2, 3) , Hiroshi Kanai 3, 2) , Harumi
Koibuchi 1) , Yasutomo Fujii 1) , Kei Konno 1) , Toshiyuki Yamada 1) and Nobuyuki Taniguchi 1)
1) Department of Clinical Laboratory Medicine, Jichi Medical University, Tochigi, Japan
2) Graduate School of Biomedical Engineering, Tohoku University, Sendai, Japan
3) Graduate School of Engineering, Tohoku University, Sendai, Japan
Aim: The aim of the present study was to investigate the utility of atherosclerotic evaluation of brachial
and radial arterial wall elasticity (AWE) by the phased tracking method in patients with type 2 diabetes
mellitus (T2DM).
Methods: This hospital-based cross-sectional study included 244 patients with T2DM(males: 61%,
mean age: 60 years). The patients were classified by vascular manifestations including a history of
celebrovascular disease, coronary artery disease, diabetic retinopathy and nephropathy. The smoking
habits, body mass index (BMI), blood pressure (BP), hemoglobin A1c, serum low-density lipoprotein,
high-density lipoprotein, triglyceride, creatinine, and brachial and radial AWE were recorded.
Differences of brachial and radial AWE by the progression of vascular manifestations were analyzed.
Results: A total of 121 patients had vascular manifestations (50%). In the univariate analyses, a
significant correlation was detected between brachial AWE and BMI (r = 0.30), systolic BP (r = 0.41),
creatinine (r = 0.19), high-density lipoprotein (r = -0.17) and triglyceride (r = 0.14). Radial AWE had a
significant correlation with BMI (r = 0.38), systolic BP (r = 0.48), creatinine (r = 0.19) and triglyceride
(r = 0.18). These correlations were not largely altered in patients with or without vascular
manifestations. Brachial and radial AWE of patients with vascular disease were significantly higher than
those of patients without vascular manifestations (brachial AWE, 791 ± 236 kPa [Pascal] and 682 ± 215
kPa, p < 0.01; radial AWE, 752 ± 245 kPa and 633 ± 209 kPa, p < 0.01). Receiver operating
characteristic curve of boundary value of brachial and radial AWE revealed an area under the curve to
identify the patients with vascular manifestations (brachial AWE, 0.65, p

No. 68 (PP 1)
The expressions of O 6 -methylguanine DNA methyltransferase and epidermal
growth factor receptor on ganglioglioma: A clinicopathologic and
immunohistochemical study
I-Wei Chang 1,2 , Jui-Wei Lin 3
1
Department of Pathology, E-DA Hospital/I-Shou University
2
Institute of Biotechnology and Chemical Engineering, I-Shou University
3
Department of Pathology , Kaohsiung Chang Gung Memorial Hospital, Taiwan
Background: Ganglioglioma (GG) is an uncommon brain parenchymal neoplasm. Although most cases
have indolent clinical behavior, a subgroup of GGs with anaplastic change in the glial component has
aggressive behavior and less favorable outcome. O6-methylguanine DNA methyltransferase (MGMT)
is a DNA repair protein that removes mutagenic and cytotoxic adducts from O6-guanine in DNA. Lack
of MGMT protein expression immunohistochemically is related to drug responses in patients of
malignant glioma treated with alkylating agents. EGFR is the most frequently amplified gene in
glioblastoma and associated with tumor invasiveness, angiogenesis, poor survival, and resistance to
radiation therapy. To elucidate the relationship between the statuses of the MGMT as well as EGFR
proteins and the tumor grading and prognosis, we conduct this study.
Methods: Clinicopathologic and immunohistochemical study of nine cases was performed.
Results: This series included four men and five women with a mean age of 21.4 years at first surgery.
Immunohistochemically, the MGMT and EGFR protein expressions tended to be more intensive in
anaplastic GG, corresponding to the worse prognostic predictive value of 3+ MGMT and 2+ EGFR
staining in glial cell component, as well as 1+ EGFR staining in neuronal cell components.
Conclusions: The series showed that MGMT and EGFR protein expressions tended to be more
intensive in anaplastic ganglioglioma.
- 121 -

No. 69 (PP 2)
Pathological characteristics of HER2/neu-amplified gastric carcinomas
M. Bamba, H. Sugihara, S. Takemura, K. Fukuda, M. Masuyama, T. Shigematsu and R. Kushima
Depts. Pathology (MB&ST), Surgery (KF&MM) and Gastroenterology (TS), Saiseikai Shiga Hosp.
Dept. Pathology, Shiga Univ. Med. Sci. (HS)
Dept. Pathology, National Cancer Center Hosp., Japan
Recently, HER2/neu becomes a molecular target for treatment of gastric carcinomas with a monoclonal
antibody trastuzumab. However, evaluation of HER2/neu amplification and overexpression are not
easy because of their intratumoral heterogeneities that are more frequent in gastric carcinomas than in
breast carcinomas. In this study, using gastric carcinomas that showed HER2/neu-amplification at
least in part, we immunohistochemically analyzed heterogeneities in HER2/neu expression.
This study was based on the analysis of 72 surgically resected gastric carcinoma cases (35 early and 42
advanced lesions). First, fluorescence in situ hybridization (FISH) was performed in all 77 lesions.
Then, H&E and immunohistochemical stainings with the monoclonal antibodies MUC5AC, MUC6,
CD10, MUC2, CDX2, HER2/neu and p53 were carried out in the HER2/neu-amplified cancers.
Twelve out of 62 informative cancers were HER2/neu-amplified (19.4%). HER2/neu overexpression
was homogeneous in 2 cancers, heterogeneous in 9 cancers and negative in 1 cancer. Seven, 5 and
none of the cancers were differentiated type, mixed differentiated and undifferentiated type and
undifferentiated type, respectively. The depth of invasion was the mucosa in 5 cancers, the submucosa
in 1 cancer and the muscularis propria or deeper in 6 cancers. p53 overexpression was diffuse in 8
cancers, focal in 1 cancer and negative in 3 cancers. Phenotype was intestinal in 5 cancers, mixed
gastric and intestinal in 6 cancers and null in 1 cancer.
HER2/neu-amplified gastric carcinomas often showed heterogeneous overexpression for HER2/neu
(75%) and diffuse overexpression for p53 (66.7%) and did not show pure gastric phenotype or
undifferentiated type in the present study.
- 122 -

No. 70 (PP 3)
Poorly differentiated squamous cell carcinoma of the nipple; A uniqu case for
marked exophytic growth, but little invasion with neuroendocrine differentiation
Naoki Hosaka, Susumu Ikehara, Hakuo Takahashi
Department of Pathology, Kansai Medical University-Kori Hosptal, Department of stem cell disoders,
Department of Clinical Sciences and Laboratory Medicine, Kansai Medical University, Osaka, Japan
A 73-year-old woman showed marked exophytic growth of a tumor (25 × 23 × 14 mm) of the nipple
over a period of 2 months. Histologically, numerous tumor nodules with no apparent keratinization were
observed in the exophytic lesion. The tumor cells also showed little invasion to the dermis and no
metastasis to the axillary lymph nodes (LN). The tumor cells were immunohistochemically positive for
cytokeratins (CKs; AE1/AE3 and 34bE12), epithelial membrane antigen (EMA) and p53, but negative
for Ber-EP4 and human papilloma virus (HPV). The MIB-1 index was 56%. Some tumor cells were also
positive for some neuroendocrine markers, and showed some tonofilaments and neurosecretory granules
in the cytoplasm under electron microscopy. We made the differential diagnosis of mammary ductal
carcinoma, basal cell carcinoma (BCC), Paget’s disease, and neuroendocrine carcinoma including
Merkel cell carcinoma. The final diagnosis was poorly differentiated squamous cell carcinoma (SCC)
showing exophytic growth with neuroendocrine differentiation (ND) in the nipple. To our knowledge,
although only five cases of Bowen’s disease have been reported in the nipple, such a unique SCC has
not been reported previously.
- 123 -

No. 71 (PP 4)
Typing of amyloidosis by proteome analysis
Masayoshi Tasaki 1, 2 , Mitsuharu Ueda 3 , Konen Obayashi 3 , Hiroyuki Hata 2 , Hirofumi Jono 3 , Genki
Suenaga 1 , Su Yu 1 , Satoru Shinriki 3 , Taro Yamashita 1 , Yukio Ando 1
1 Department of Neurology, Graduate School of Medical Sciences, Kumamoto University, Kumamoto,
Japan
2 Department of Immunology and Hematology, Division of Health Sciences, Faculty of Life Sciences,
Kumamoto University, Kumamoto, Japan.
3 Department of Diagnostic Medicine, Graduate School of Medical Sciences, Kumamoto University,
Kumamoto, Japan
[Background]Amyloidosis is one of the protein conformational disorders that normally soluble proteins
accumulate insoluble amyloid fibrils, with the result being severe organ dysfunction. It is documented
that 27 different amyloidogenic proteins. While pathologists identify the amyloid precursor protein by
immunohistochemistry (IHC) using antibodies against known amyloidogenic proteins, the results may
be inconclusive owing to the loss of epitope or small amounts of amyloid deposits, unknown
amyloidogenic protein. In this study, we investigated the usefulness of proteomic analysis for amyloid
typing.
[Methods]We used formalin-fixed and paraffin-embedded (FFPE) tissue sections for identifying
amyloid precursor protein in amyloid-laden specimen using LC-MS/ MS. We attempted retrospective
analysis of 20 amyloid-laden specimens (AL, FAP, SSA AA and DRA) to detect amyloidogenic protein.
Therefore, we analyzed amyloid precursor protein of 9 amyloid specimens including cases unidentified
by IHC in a prospective study.
[Result] In the retrospective study, we detected amyloidogenic protein from FFPE materials in some
type of amyloid using LC-MS/ MS. The concordance rate with IHC was 95% (19/ 20). Therefore, in
cases IHC was difficult to determine, we also identified amyloidogenic protein in a prospective study.
[Conclusion] LC-MS/ MS have a potential tool to identify amyloidgenic protein from FFPE tissue. This
method is useful for diagnosis of amyloidosis, even cases that are difficult to diagnose by IHC and in
rare cases of amyloidosis.
- 124 -

No. 72 (PP 5)
The morphological change after molecular targeting therapy of malignant brain
tumor
Tatsuo Sawada 1 , Yoshinori Takekawa 2 , Saori Okamoto 3 , Takashi Maruyama 3 , and Noriyuki
Shibata 1
1 Department of Pathology, Tokyo Women’s Medical University
2 Department of Neurosurgery Tokyo Women’s Medical University
3 Department of Surgical Pathology Yokosuka Municipal Hospital, Japan
(Background) The molecular targeting agents that are currently available for various malignant
neoplasms. The effects of molecular targeting agents are divided into two subgroups, oncogene
addiction (OA) and non-oncogene addiction (NOA). Bevacizumab, anti-vascular endothelial growth
factor antibody is one of major agents of non-oncogenic addition although the therapy is permitted only
experimentally in Japan. Malignant glioma, especially, glioblastoma (GB), and highly vascularized brain
tumors are thought to be the one of the attractive targets for anti-angiogenic therapies. To elucidate
morphological changes of tumor after NOD therapy is important to evaluate the effects of the drug. So
we examined the morphological change of malignant glioma after Bavacizumab therapy using 3 autopsy
cases.
(Materials and Methods) Three cases in men were used. The range of age is from 37 to 64yrs. These 3
cases received Bemacizumab therapy, and radiation therapies were combined. The microscopic slides of
surgical resected and autopsied specimen were compared using HE and immunohistochemistry (IHC)
for vascular endothelial growth factor, EGF A and C, vascular endothelial growth factor receptor, Fltand
Flk-1, platelet derived growth factor (PDGF), CD31, CD34, smooth muscle cell actin (SMA) and
aquaporin 1 and 4. And 5 cases of GB are examined for control.
(Results) No significant change of morphology of tumor cells is identified after Bevacizumab therapy,
but the morphology of neovasculature is different from conventional GB. A few glomeruloid
appearances and hyperplasia of SMA positive cells are identified. On IHC, the reactivity for VEGF A
was decreased but VEGF-C and Flt-1 were preserved.
(Conclusion) The morphological difference between with and without administration of Bemasizumab
in neovasculature suggests decreased expression of VEGF-A.
- 125 -

No. 73 (PP 6)
Clinicopathological analysis of asbestos-related diseases presenting asbestos bodies
in routine cytology specimens
Makihara K 1) , Kanazawa S 1) , Yoshida T 1) , Sosogi A 1) , Matsuki Y 2) , Shimajiri S 3) , Yamasaki K 4) ,
Hamada T 1)
1) Dept. Surgical Pathology and Asbestos Disease Center, Kyushu Rosai Hospital,
2) Dept. Surgical Pathology, Ootemachi Hospital,
3) Dept. Surgical Pathology, Univ. Occupational and Environmental Health, Japan
4) Dept. Respiratory Medicine, Univ. Occupational and Environmental Health , Japan
In Japan, asbestos-related diseases are predicted to increase in number within a few decades probably
due to the retarded political and legal acts for protection from the toxicity of asbestos fibers (AF).
Asbestos body (AB) is rarely encountered in the cytology smears particularly of routine specimens even
in the patient presenting asbestos-related respiratory symptoms. In our surgical pathology file, ABs
could be detected in 5 cases out of 1253 cases (0.40%), 2496 specimens in which cytological
examinations were performed in respiratory specimens. The smears included the specimens of sputum,
bronchial brushing, bronchoalveolar lavage (BAL), bronchial aspirate and tissue-imprint. All patients
were 58 to 72 (mean: 67.0) years-old Japanese male. Among the 5 cases, ABs were identified in the
smears of sputum (1/5, 20%), BAL (3/5, 60%), and tissue-imprint (1/5, 20%). A large number of ABs
were found in one case (case #1) who died due to respiratory failure by asbestosis and submit autopsy
(case report presented in the 59 th National Congress of JSLM). Briefly, he had the history of
occupational asbestos-exposure for 13 years. Over 100 asbestos bodies were counted in one H&E
sections of the lung in histological examination. Case #2 was histologically diagnosed as
pneumoconiosis after cytological identification of AB in BAL fluid. Case #3 was pulmonary nodular
amyloidosis and AB was incidentally noticed in the BAL smears. In the case #4, BAL was performed
for the examination of interstitial pneumonia associated with rheumatoid arthritis, and AB was
cytologically observed in the fluid. In another case #5, AB was found in the tissue-imprint smears of the
lung of VATS specimens resected for metastatic tumor from rectal cancer. Tissue specimens were
obtained in the cases #1 and #5 in which analyses of AB and AF will be carried out after extraction from
formalin fixed or paraffin embedded tissues.
- 126 -

No. 74 (PP 7)
Neuropathological findings in a case of amyotrophic lateral sclerosis resembling
CIDP
Masaru Yoshioka (1), Hidehiko Konno (1), Toshiaki Takahashi (1), Hiroyasu Tanaka (1), Hiroshi
Onodera (1), Maki Tateyama (2)
(1) Department of Neurology, National Hospital Organization Nishitaga Hospital
(2) Department of Neurology, Tohoku University School of Medicine, Japan
Aim: We present a case fulfilling diagnostic criteria of chronic inflammatory demyelinating
polyradiculoneuropathy (CIDP) with a progressive course refractory to immunosuppressive therapy, and
showing autopsy findings indicative of amyotrophic lateral sclerosis.
Methods: A clinicopathological case report.
Results: A patient, 62 year-old male, developed motor weakness, muscle atrophy, and sensory deficits in
both upper and lower limbs in a progressive course. On examinations, cerebrospinal fluid showed
protein-cellular dissociation, neurophysiological studies revealed reduced motor and sensory nerve
conduction velocities with prolonged distal latencies in multiple nerves. Biopsy of sural nerve showed
segmental demyelination in teased fibers, fulfilling diagnostic criteria of definite CIDP. Systemic motor
weakness and muscle atrophy progressed after repeated intravenous immunoglobulin infusions and
steroid pulse therapies. Patient died of respiratory failure after the disease duration of 3 years and 5
months. Autopsy revealed diffuse loss of myelinated fibers in anterior and posterior roots. Cellular loss
of posterior root ganglia was observed as well. Sciatic nerve showed demyelination and perineurial
edema. Anterior horn cells were massively reduced and pallor of lateral columns was found along the
whole levels of spinal cord. Iliopsoas muscle and diaphragm exhibited marked neurogenic atrophy.
Some brainstem nuclei showed neuronal loss.
Conclusions: The present case suggests that an association of amyotrophic lateral sclerosis may be a
factor for
therapeutic refractoriness in some patients with polyneuropathy resembling CIDP. Causal relationship
between demyelinating neuropathy and loss of anterior horn cells is uncertain.
- 127 -

No. 75 (PMB 1)
Synthesis of cDNA and Detection of Gene Fragment of Protein Disulfide Isomerase
Family a Member 4 (PDIA4) in Bone Marrow Cell
Stefanus Lembar
Atma Jaya Medical Faculty Indonesia
Background: Protein disulfide isomerase (PDI) is a member of the thioredoxin super-family that
present in the endoplasmic reticulum (ER) in eukaryotic cells. PDI catalyzes of disulfide bond formation,
reduction, or isomerization of newly synthesized proteins in the lumen of the ER. PDI confers resistance
to apoptosis under hypoxic conditions. Hypoxia is physiologically very important in the mechanism of
ER stress. Several clinical studies indicate that tumor hypoxia predicts the decline of local control,
increased distance metastasis, and decreased survival of tumor cells. PDIA4 or also known as ERp72 is
a PDI family member that expressed on bone marrow cells and mammary gland. Low expression of
PDIA4 as a result of ER stress on the condition of hypoxia may play a role in the process of tumor
metastasis. Therefore, detection and amplification of PDIA4 gene needs to be done to study the
mechanism and changes in sequences of PDIA4 that may contribute to tumor metastasis.
Objective: To synthesize of cDNA and detection of PDIA4 gene fragment in bone marrow cell.
Methods: cDNA was synthesized from bone marrow cell total RNA by RT-PCR using a commercial kit.
Qualitative and quantitative analysis of cDNA performed using the NanoDrop. Detection of PDIA4
gene fragment by cDNA amplification using conventional PCR. PCR products visualized using 1 %
agarose gel electrophoresis.
Results: Quantitative analysis of cDNA showed that the cDNA has a concentration of 383 ng/μL.
Amplification of cDNA with primers F-full mPdia4 and R-full mPdia4 successfully carried out with the
primers concentration of 100 nM and annealing temperature of 50 °C. PCR products were demonstrated
by visualization of DNA bands on agaorse gel electrophoresis with a size of about 1925 bp.
Conclusion: cDNA have been successfully synthesized from bone marrow cell total RNA and amplified.
Furthermore, DNA sequencing will be conducted to verify the suitability of the PDIA4 gene fragment
sequences.
- 128 -

No. 76 (PMB 2)
Diagnostic strategies lynch syndrome - role of immunohistochemistry and germline
mutation screening -
Mohd Nizam Zahary 1 , Gurjeet Kaur 2 , Muhammad Radzi Abu Hassan 3 , Ahmad Shanwani Mohd
Sidek 4 , Harjinder Singh 5 , Sharifah Emilia Tuan Shariff 6 , Ravindran Ankathil 1 .
1
Human Genome Centre, School of Medical Sciences, Universiti Sains Malaysia Health Campus,
Kubang Kerian, Kelantan, 2 Institute For Research in Molecular Medicine, Universiti Sains Malaysia,
3 4
Internal Medicine Department, Hospital Alor Setar, Kedah, Surgery Department, Hospital Raja
Perempuan Zainab II, Kota Bharu, Kelantan, 5 Department of Medicine, Hospital Queen Elizabeth, Kota
Kinabalu, Sabah, 6 Department of Pathology, School of Medical Sciences, Universiti Sains Malaysia
Health Campus, Kubang Kerian, Kelantan, Malaysia
Lynch syndrome, caused by a germline mutation in a mismatch repair gene and associated with tumors
exhibiting microsatellite instability (MSI), is characterized by an increased risk of colon and
extracolonic cancers. Evaluation of MSI through immunohistochemistry of the 4 MMR proteins
followed by molecular genetic testing is necessary for accurate diagnosis of Lynch syndrome. We
identified 47 patients with suspected Lynch syndrome for whom we aimed to analyze the protein
expression profile of MMR protein, MLH1, MSH2, MSH6 and PMS2 using immunohistochemistry and
to screen for germline mutations in MLH1, MSH2, MSH6 and PMS2 gene. MMR protein expression was
immunohistochemically analyzed on paraffin blocks of resected colorectal specimens. DNA extracted
from blood were screened for germline mutations in 19 exons of MLH1, 16 exons of MSH2, 10 exons of
MSH6 and 15 exons of PMS2 gene using dHPLC and followed by DNA sequencing. Absent expression
of any MMR protein was observed in 14/47 patients (29.8%). Germline mutation of MLH1 gene was
identified in 2/47 patients (4.2%) consisted of c.1851_1853delGAA (Lys618del) but MLH1 promoter
polymorphism -93G>A was frequently identified (32/47, 68%). Germline mutation in MSH2 gene was
identified in 6/47 patients (12.8%) consisted of a novel mutation, c.2005G>C (Gly669Arg), c.142G>T
(Glu48Stop), c.2701G>A (Glu901Lys) and splice-site mutation, c.2006-6T>C which was adjacent to the
exon 13 of MSH2 gene. Germline mutation of MSH6 was also identified in 1/47 patients consisted of a
novel mutation, c.3885_3891delTAAAAGC (Lys1296MetfsX29) and mutation in PMS2 gene was
identified in 2/47 (4.2%) patients namely c.59G>A (Arg20Gln), c.1408C>T (Pro470Ser), c.1454C>A
(Thr485Lys), c.1621A>G (Lys541Glu) and c.2324A>G (Asp775Ser). Immunohistochemistry proved to
be a good pre-screening tool before germline mutation analysis of MMR genes. Immunohistochemical
profiling of protein expression and germline mutation screening of MMR genes together are important
in differential diagnosis of Lynch syndrome from familial colorectal cancer.
- 129 -

No. 77 (PMB 3)
Corneodesmosin (MHC S) Gene Polymorphism in Mongolian Psoriasis Patients
Ch.Tserendulam 1,3 , B.Munkbat 1,2 , M.Tomizawa 2 , G.Oyungerel 1 , G.Tamiya 2 , Sh.Myagmarjav 4
H.Inoko 2 , N.Munkhtuvshin 1
1
Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
2
Depart. Molecular Life Science, Tokai Uni. School of Medicine, Isehara, Kanagawa, Japan
3
Jargalan Hospital Laboratory, Khuvsgul, Mongolia
4
Nat’l Center for Dermatology and Mycology, Ulaanbaatar, Mongolia
Objectives: The aim of this study was to examine the potential pathogenic linkage of the MHC S gene
polymorphism in a Mongolian psoriasis population, known to have very strong association between
psoriasis vulgaris and the HLA – Cw6 allele. Eight SNP in the MHC S gene at positions +614, +619,
+1118, +1215, +1236, +1240, and +1372 were analyzed by direct sequencing.
Materials and Methods: A total of 101 unrelated Mongolian patients (50 male,51 female) with
psoriasis vulgaris (PV), hospitalized at the Nat’l Centre for Dermatology and Mycology, UB, Mongolia
and 117 healthy controls were investigated in this study. Genomic DNA were prepared from the freshly
collected peripheral blood using DnaQuick II kit. The PCR amplifications were carried out in a
GeneAmp PCR system 9700 thermal cycler (PE Applied Biosystems). DNA sequencing of the amplified
PCR products was performed on the ABI Prism ® 3700 DNA Analyzer.
Results: We have found a strong association between Mongolian psoriasis and a nucleotide substitution
at position +1236, which result in amino acid exchange Ala351Ser. The observed homozygotes for the
more common allele T (Ser) at +1236 was significantly higher (ORHOM = 2.775, Pc

No. 78 (PMB 4)
Study of essential oil for anti-allergy effect in murine asthma model
Tomoe Ueno Iio, Misako Shibakura, Michinori Aoe, Tomoko Hyoda, Mihoko Kohara, Naohisa
Nakagawa, Takako Nakahara, Mikio Kataoka.
Graduate School of Health Sciences, Okayama University., Japan
Background: The essential oils are used for aromatherapy classified into alternative medicine.
However, there are few evidences of the action mechanism of essential oil for the body. To elucidate
the possibility of the prevention of an allergic disease by the aromatherapy, we investigated the effects
of essential oil using murine asthma model.
Methods: Female BALB/c mice of 8-9 weeks were divided into a control mice (A), an asthma mice (B),
the essential oil (Lavandula angustifolia) treated-asthmatic mice (C) and (D). To make asthma mice, B,
C, and D were sensitized by intraperitoneal injection of Ovalbumin (OVA) at day 0 and 14, and
challenged via airways with OVA on Days 28, 29, 30. After 48 to 72 hours later of last OVA challenge,
airway resistances were assessed. The cell numbers, cytokine levels in BAL fluid, and serum
OVA-specific IgE levels were measured. The mucus-containing cells were counted in periodic
acid-Schiff (PAS) stained sections. For C and D, 5 µl and 20 µl of essential oil were applied for 20
min from day 14, 5days of week.
Results: The airway resistance significantly increased in B, C and D. The total cell numbers were
significantly reduced in C and D compared with B. The eosinophil numbers were significantly
reduced in D compared with B. IL-13 levels in BAL fluid were significantly reduced in group D
compared with group B. The serum IgE levels were increased in each group except for A. There
were significant reductions of the PAS positive cells in C and D compared with B.
Conclusion: Treatment with Lavender essential oil of asthma mice decreased the number of total cells
and eosinophils, IL-13 levels in BALF, and bronchoalveolar PAS positive cells. These results are
indicating that aromatherapy may have possibility to be used for prevention of allergic inflammation,
such as asthma.
- 131 -

No. 79 (PMB 5)
TGF-β-neutralization enhances cytarabine-induced apoptosis in AML cells in the
bone marrow microenvironment
Yoko Tabe 1,2 , Linhua Jin 1 , Yasuhito Hatanaka 1 , Takashi Miida 1 , Michael Andreeff 2 , Marina
Konopleva 2
1, Department of Clinical Laboratory Medicine, Juntendo University School of Medicine, Tokyo, Japan
2, Department of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, Texas,
USA
Aim: Hypoxia and interactions with bone marrow mesenchymal stromal cells (MSC) have emerged as
essential components of the leukemic microenvironment in promoting leukemia cell survival. High
levels of transforming growth factor beta 1 (TGF-β1) produced by stromal cells regulate cell
proliferation, survival, and apoptosis, depending on the cellular context. In this study, we investigated
the anti-leukemic effects and molecular mechanisms of action of TGF-β neutralizing antibody 1D11
under hypoxic conditions. We further investigated the anti-leukemic efficacy of 1D11 combined with
CXCR4 antagonist plerixafor in the in vivo leukemia models.
Design and Methods: The antileukemic effects and molecular mechanisms of action of monoclonal
pan-TGF-β-neutralizing antibody 1D11 were determined utilizing flow cytometry, western blot, qPCR,
and siRNA technology. The in vivo efficacy of 1D11 combined with CXCR4 antagonist Plerixafor was
investigated in the Baf3/ITD model.
Results: RhTGF-b1 induced accumulation of acute myeloid leukemia (AML) cells in a quiescent G0
state under MSC co-culture conditions, which was abrogated by 1D11. TGF-β1 upregulated p21
expression and the TGF-β target leukemia inhibitory factor (LIF) gene in AML cells which in turn
promoted pro-survival phosphorylation of Stat3. 1D11 blocked these effects and enhanced
Ara-C-induced apoptosis of AML cells in hypoxic and in normoxic conditions. Treatment with 1D11
combined with Plerixafor and Ara-C decreased leukemia burden in an in vivo leukemia model.
Conclusions: Blockade of TGF-β by 1D11 and abrogation of CXCL12/CXCR4 signaling may enhance
the efficacy of chemotherapy against AML cells in the hypoxic BM microenvironment.
- 132 -

No. 80 (PMB 6)
Introduction of HCV Quantification as a Diagnostic Tool in Mongolia: Significance
and Lessons Learned
Batmunkh Sayabold 1 , Nemekhbaatar Lkhaasuren 2 , Jazag Amarsanaa 1,5 , Jigjidsuren Chinburen 1,3 ,
Oidov Baatarkhuu 1,2 , Byambaa Suvd 1 , Tseveg Tsogtkhishig 2 , Bira Tsatsralt-Od 1,4 , Batmunkh
Munkhbat 5
1) Mongolian Association for the Study of Liver Diseases, Mongolia; 2) Health Sciences University of
Mongolia, Mongolia; 3) National Cancer Center, Mongolia; 4) National Center of Communicable
Diseases, Mongolia; 5) Institute of Medical Sciences, Mongolia
Background: The 17% of Mongolian population is infected with HCV, which makes the country one of
the top HCV infected countries in the world. Though HCV quantification method was developed almost
a decade ago, it was first introduced in Mongolia only in 2009.
Methods: Total of HCV 212 patients were enrolled in the study. HCV quantification was performed at
Happy Veritas clinical laboratory by Taqman real-time PCR method using ABI7000 from Applied
Bio-systems, USA. Commercially available HCV quantification kit was used. Anti-HCV was purchased
from ACON laboratories, USA.
Results: Out of 212 patients who had HCV quantified, 35 were also checked for HCV antibody. Patients
were divided in high viral load>2 million HCV copies/ml, and low-intermediate

No. 81 (PMB 7)
Development of Thermostabilised MultiplexPCR For Detection of Salmonella
Enteritidis and Vancomycin-Resistant enterococci (VRE)
Tan Ka Liong a , Chan Yean Yean b
a Department of pharmacology, b Department of microbiology and parasitology, School of Medical
Sciences, Universiti Sains Malaysia, Malaysia
Background: Enterococcus spp. is one of prominent nosocomial pathogens that cause clinical
infections. CDC has reported a remarkable increase in the occurrence of vancomycin-resistant
enterococci (VRE) in hospitals. VRE and SalmonellaEnteritidis (SE) have been reported in poultry
samples and transmission to human. We aim to develop a thermostabilized multiplex PCR, a
DNA-based cold chain free test for identification of VRE and SE.
Methods: VRE Multiplex PCR that detects 6 genes simultaneously was developed in previous study.
The Salmonella Enteritidis virulence gene detection was incorporated. Several conditions (e.g.
annealing temperature, premix buffer selection, concentration of Taq polymerase and concentration of
enzyme stabilizer) of PCR were optimized.
Results: In multiplex PCR, optimum annealing temperature of 64.8oC and commercial premix C buffer
for PCR reaction were determined. 75% of Taq polymerase and 3% enzyme stabilizer appeared to be
optimum in thermostabilized multiplex PCR.
Conclusion: The DNA based method was successfully optimized. This detection can be used to
facilitate surveillance in poultry and human samples.
- 134 -

No. 82 (PMB 8)
Analysis of Fas exon 6 splicing and gene expression of transcription factors in
various human organs and lymphocyte subsets
Ryo Uozumi 1 , Hitoshi Shibuya 1 , Akio Shigematsu 1 , Kazuhiko Matsuno 1 , Chikara Shimizu 1 ,
Seiichi Kobayashi 2 .
1 Department of Clinical Laboratory, Support of Clinical Practice, Hokkaido University Hospital,
2 Department of Medical Laboratory Science, Faculty of Health Sciences, Hokkaido University, Japan
Exclusion of Fas gene exon 6 by alternative splicing of the primary RNA transcript of the
apoptosis-inducing Fas gene produces a soluble isoform. The membrane Fas (mFas) isoform induces
apoptosis, while the soluble Fas (sFas) isoform prevents apoptosis. Serum levels of sFas in patients with
some autoimmune diseases and with cancer are higher than healthy individuals, and clinically correlate
with disease activity or tumor mass size. However, the regulatory mechanism for Fas gene splicing
remains unknown. Several pre-mRNA splicing regulators for Fas gene were reported, but these studies
only targeted extrinsic Fas minigene introduced into cell lines. Here, by quantitative real-time RT-PCR
analysis, we have investigated whether candidates of transcription factors including Hu antigen R
(HuR), T-cell intracellular antigen 1 (TIA-1) and TIA-1 related proteins (TIAR) actually regulate
intrinsic Fas gene splicing in various human organs and lymphocyte subsets. Each gene expression
was different in organs, but there was a poor correlation in gene expression between Fas and
transcription factors. We next analyzed
gene expression by CD45RA+ or CD45RO+ peripheral lymphocytes from healthy donors. Although
sFas and mFas gene expression by these subsets were significantly different, no correlation between
Fas, HuR, and (TIA-1+TIAR) gene expression was obtained. These results suggested that transcription
factors reported should not necessarily function as regulators for intrinsic Fas gene splicing.
- 135 -

No. 83 (PMB 9)
The genetic basis of Congenital Hyperinsulinism in Japan
Kumiko Ohkubo 1)2) , Mariko Nagashima 1) , Tomoe Matsuzaki 2) , Makiko Yuki 2) , Hironobu
Kawashima 2) , Akira Matsunaga 1)2)
1) Department of Laboratory Medicine, Faculty of Medicine, Fukuoka University, Japan
2) Department of Clinical Laboratory, Fukuoka University Hospital, Japan
Objective Congenital Hyperinsulinism (CHI) is a disorder of glucose metabolism by dysregulated
secretion of insulin from pancreatic β-cells. This disease is associated with mutations of ABCC8 and
KCNJ11, which are genes of two subunits of the pancreatic β-cell ATP sensitive potassium channel.
Patients and Methods In 31 Japanese CHI patients, all exons of ABCC8 and KCNJ11 were analyzed by
PCR and direct sequencing. Multiplex Ligation-dependent Probe Amplification (MLPA) was also
applied to analysis of copy numbers in the region of ABCC8.
Results We found mutations in 22 patients out of 31 patients. The 19 patients showed mutations in
ABCC8 and 3 patients showed those in KCNJ11. The mutations of ABCC8 in 19 patients contained 7
nonsense mutations, 8 missense mutations, 3 aberrant-splicing mutations and 1 one base deletion
mutation. These mutations distributed broad in ABCC8, although some missense mutations existed
around two nucleotide-binding domains and membrane domains. The mutations of KCNJ11 in 3
patients contained 2 missense mutations, and each one of one base deletion and two base deletion
mutations. One patient having KCNJ11 mutation was a compound heterozygote of missense and one
base deletion mutations. All of the patients having missense mutations in ABCC8 were heterozygotes
and responsive to diazoxide or octreotide to inhibit insulin secretion except one patient, who needed
operation. All of the patients showing nonsense mutations in ABCC8 underwent a subtotal or partial
pancreatectomy to control blood glucose levels. The homozyotes of ABCC8 mutations or the compound
heterozygotes of KCNJ11 mutations also had operations. Analysis by MLPA showed no abnormal
results of copy numbers in ABCC8.
Conclusion The 23 kinds of mutations reported in this study could be pathological candidates for CHI
in Japan. The genetic diagnosis for these mutations might be applicable for prediction of prognosis and
improvement of management in CHI patients.
- 136 -

No. 84 (PMB 10)
A rapid and automated detection system for BRAF mutations in malignant
melanoma
Masaru Ide, Shinichi Koba, Yumi Nagano, Naoko Aragane-Sueoka, Akemi Sato, Takuya Inoue,
Naomi Kobayashi, Noriyuki Misago, Yutaka Narisawa, Shinya Kimura and Eisaburo Sueoka
Faculty of Medicine, Saga University, Japan
BRAF gene mutations have been reported in 30-50% of malignant melanoma patients. Because of the
difficulty of early diagnosis of the disease, poor prognosis was observed in far advanced melanoma
patients. However, recent development of therapeutic intervention using BRAF inhibitors showed
improved prognosis of such patients. Therefore, an accurate and rapid detection system for BRAF
mutations is desired. In addition, the clinical characteristics of the melanoma associated with BRAF
mutations in Japanese patients have not been investigated on a large scale evaluation. We recently
established quenching probe system (QP) for detection of an activating BRAF mutation V600E, and
evaluated 113 melanoma samples diagnosed in Saga University Hospital from 1982 to 2011. The QP
system includes fully automated genotyping, based on analysis of the probe DNA melting curve, which
binds the target mutated sites using a fluorescent guanine quenched probe. BRAF mutations were
detected in 54 of 115 (47%) including 51 of V600E and 3 of V600K in Japanese melanoma cases.
Among clinical subtypes of melanoma, nodular melanoma showed high frequency (12 of 15; 80%) of
the mutation followed by superficial spreading melanoma (13 of 26; 50%). Clinical stages and sex
difference were not associated with mutation frequency and mutation sites. The QP system is a simple
and sensitive method to determine BRAF V600E mutation, and will be useful tool for patient-oriented
therapy with BRAF inhibitors.
- 137 -

No. 85 (PMB 11)
Detection of Mycobacterium kansasii using amplicons generated by the COBAS
TaqMan 48 Analyzer
Arizumi Kikuchi 1 , Yuko Wakayama 1 , Takahiro Sawamura 1 , Satsuki Nakamura 2 , Shu Taga 2 , Yuji
Itoh 2 , Takeshi Seki 2 , Hiroya Sakuma 1,2 , Osami Daimaru 1,2 , Takeo Nakakita 1,2 , Shinichi Itoh 3
1 Daiyukai Second Medical and Science Research Laboratories, 2 Daiyukai General Hospital, 3 Daiyukai
First Hospital, Ichinomiya, Aichi, Japan
Mycobacterium detection using genetic techniques are rapid tests that are mainly based on nucleic acid
amplification. Many laboratories in Japan use the COBAS TaqMan 48 Analyzer (Roche Diagnostics), a
real-time PCR-based system that uses TaqMan hydrolysis probes. This system is efficient and easy to
use and has good sensitivity and specificity. However, this system can only detect the M. tuberculosis
and M. avium complex among mycobacteria. We examined methods to detect M. kansasii using the
amplicon generated by the COBAS TaqMan 48 Analyzer by real-time PCR using a hydrolysis probe.
We used the type strains of 7 mycobacterial species: M. avium, M. fortuitum, M. gastri, M. intracellular,
M. kansasii, M. marinum, and M. tuberculosis. We extracted the total nucleic acids from these type
strains and performed the COBAS TaqMan MTB Test (Roche Diagnostics) or the COBAS TaqMan
MAI Test (Roche Diagnostics) on a COBAS TaqMan 48 Analyzer. After the reaction, the amplicons
were cleaned using the High Pure PCR Product Purification Kit (Roche Diagnostics). The M. kansasii
detection primer pair was designed from the inner sequence of 16S rRNA, which was used in the
COBAS TaqMan 48 Analyzer. The detection probe used was highly specific for M. kansasii, and the
locked nucleic acids were incorporated into the probe to increase specificity. Real-time PCR was
performed using Light Cycler 1.5 (Roche Diagnostics).The primer and probe for M. kansasii detection
accepted the amplification of M. kansasii and M. gastri. M. kansasii lung disease shows the highest
response to chemotherapy, and the method for its treatment is better established than that for other
non-tuberculous mycobacterial infectious diseases. Since early and rapid detection of M. kansasii is
very important, we suggested that our method can be employed to detect M. kansasii using COBAS
TaqMan 48 Analyzer in routine laboratory work.
- 138 -

No. 86 (PMB 12)
Increased expression of mitochondrial isoenzyme of creatine kinase in
hepatocellular carcinoma cells may be associated with enhanced proliferation and
migration
Uranbileg B., Ikeda H., Yatomi Y.
Project researcher, Department of Clinical Laboratory Medicine, Graduate School of Medicine, The
University of Tokyo, Japan
Mitochondrial isoenzyme of creatine kinase (MtCK) has been postulated to be important for the
energetics of oxidative tissues, suggesting that MtCK might play crucial role(s) in malignant
transformation.
We have recently reported the increased serum MtCK activity in patients with hepatocellular carcinoma
(HCC), and suggested that MtCK could be a novel serum marker for HCC. Although we revealed the
increased MtCK mRNA expression in HCC cell lines as one of its mechanisms, its significance has been
largely unknown.
The reduction of MtCK expression in HCC cell lines, i.e., HepG2, Alex and HuH7, by siRNA led to the
decrease in cell proliferation by 31.7% in HuH7, 19.8% in HepG2, and 15.5% in Alex measured by
BrdU incorporation assay. Furthermore, the reduction of MtCK expression in HepG2, Alex, HuH7
caused the decrease in cell migration, maximum by 85.2% in HuH7, and cell invasiveness also reduced
in all three cell lines, maximally detected in HuH7 by 92.3% which was determined by cell migration
and invasion assay.
Because ASB9 (Ankyrin repeat and suppressor of cytokine signaling box protein 9) was previously
reported as a potential regulator of MtCK expression, its expression level in HCC cell lines was
investigated. It was found that ASB9 mRNA expression in all HCC cell lines examined was much lower
than that in normal liver tissue. This negative correlation between the expressions of MtCK and ASB9 in
HCC cell lines may suggest that ASB9 could be a negative regulator for MtCK expression in HCC.
In conclusion, the increased expression of MtCK in HCC cell lines may be associated with the enhanced
proliferation, migration and invasion, which consistent with MtCK is being a serum marker for HCC.
MtCK and its negative regulator, ASB9, merits consideration as a therapeutic target for HCC.
- 139 -

No 87 (PO 1)
Application for Diagnostic Tests of Facial Portraits by CCD Camera
Kazuhiro Kabe
Business Department of Medical Technology, Laboratory of Medical Technology
Medic21 Association, Japan
Aim : At daily clinical diagnostic tests, as it is very important to judge whose each specimen it is, we
can think that it is good to photograph faces in order to discriminate individuals of test subjects. I
investigated the changes by making facial portraits by a CCD Camera, by discriminating individuals and
by including simple photographing systems in personal-computers to determine whether individual
discrimination of judged specimens succeeds to be practicable.
Materials and Methods : I performed by joining CCD Camera with personal-computers, through
softwares to take in portraits, by adjusting shades of portraits to signals, by dividing into two kinds, by
letting indicate that shadowy directions were “1” and light directions were “0”, by forming a line of both
0 and 1 and by including systems that let express facial portraits by both 0 and 1 in personal-computers.
Results : As a result of expressing man’s facial portraits by shades of both 0 and 1, man’s facial portraits
formed a line of shades of both 0 and 1 and at both the same man and position, in comparing the first
time with the second time, coincided almost closely. But when the photographing position changed,
somewhat slipped out rows of both 0 and 1. And when the man also changed, somewhat slipped out
rows of that.
Conclusions : By using CCD Camera, individual discrimination by facial portraits seems to be well
practicable. Therefore, by also using cellular phones, individual discrimination seems to be well
practicable. At clinical diagnostic tests, if we introduce these CCD cameras, individual discrimination of
specimens seems to be effectively practicable by using that of this facial portrait.
- 140 -

No 88 (PO 2)
Assessment of early manifestation of anthracycline-induced cardiomyopathy in
patients with preserved left ventricular ejection fraction
Reiko Mizuno 1 , Shinichi Fujimoto 2 , Yasuyuki Okamoto 1
1 Central Clinical Laboratory, Nara Medical University, 2 Center for Education Development, Nara
Medical University, Japan
Aim: Cardiac side effect of anthracycline limits its potential utility. Recent studies have reported that
latent anthracycline-induced cardiomyopathy already exists even in patients with preserved left
ventricular (LV) ejection fraction (EF). We investigated LV mechanical dyssynchrony in
anthracycline-induced cardiomyopathy with preserved LVEF.
Methods: We studied forty-seven patients with non-Hodgkin’s lymphoma who were previously treated
with anthracycline repeatedly. All patients had no history of cardiac disease and showed preserved
LVEF (≥ 60%). Myocardial strain rate obtained with speckle tracking was used to assess ventricular
synchrony. Time to regional peak strain rate was measured in 12 LV segments from the apical 4- and 2-
chamber views and standard deviation (SD) in all 12 segments was measured as an estimate of LV
dyssynchrony. Also the relationship between SD and LVEF was assessed. Thirty-eight healthy
volunteers served as controls.
Results: Although LVEF was preserved in all patients, it was significantly decreased compared with
controls. SD of time to peak strain rate was significantly greater in patients than controls. SD was
negatively correlated with LVEF in patients.
Conclusion: This study suggests that LV mechanical dyssynchrony exists and causes latent LV
dysfunction in patients with anthracycline-induced cardiomyopathy with preserved LVEF.
- 141 -

No. 89 (PO 3)
Influence of masticatory dysfunction on antioxidant capacity in rat model
Maki Tanaka, Kageaki Kuribayashi, Daisuke Kobayashi, Naoki Watanabe
Department of Clinical Laboratory Medicine, Sapporo Medical University School of Medicine, Japan
Mastication is an essential function to maintain homeostasis of the animal. Disturbance in mastication
induces physical and psychological stresses. It has been elucidated that level of free radical, such as
superoxide, increases following physical and psychological stresses leading to various disorders,
including cancer and gastrointestinal ulcer. In this study, we investigated whether masticatory
dysfunction causes oxidative stress by changing feeding style from solid to liquid meal in rat model.
Neutrophils were collected from rats fed with solid or liquid meal, respectively, and superoxide
production was measured by cytochrome c reduction method. Serum superoxide dismutase activity was
evaluated using electron spin resonance spin-trapping technique. As a result, level of superoxide
production was higher in rats fed with liquid than solid meal. Furthermore, serum superoxide dismutase
(SOD) activity was lower in the former group. Decrease in serum SOD activity sustained from day 21 to
day 84 after meal was changed from solid to liquid meal. In rats fed with liquid meal, serum SOD
activity recovered to normal level 7 days after liquid meal was reversed to solid meal. These results
show that oxidative stress continues when mastication is disturbed. This study suggests that chewing
harmless solid materials, such as gum, may help avoiding unnecessary oxidative stress in patients
treated with liquid nutrition.
- 142 -

No. 90 (PO 4)
The role of Rb2/p130 and PP2A interaction in ATRA treatment induced growth
arrest of ovarian carcinoma cells
M.Oyundelger 1,3 , P.Enkhtsetseg 1,2 , Soprano DR 2 , Soprano KJ 2 , N.Munkhtuvshin 1
1
Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
2
Department of Microbiology and Immunology, Temple University School of Medicine, Philadelphia,
3 CEO Laboratory, Monolab Co., Ulaanbaatar, Mongolia
Objectives: Further characterization of the interaction between Rb3/p130 and PP2A upon All-trans
retinoic acid (ATRA) treatment as it has been shown to arrest the growth of the CAOV3 ovarian
carcinoma cells in the GO/G1 phase and to elevate levels of Rb2/p130 protein, a member of the
retinoblastoma family of tumor suppressors. Materials and Methods: CAOV3, was obtained from
theAmerican Culture Collection (Rock-ville, MD), Anti-PP2Ac, Anti-Rb2/p130, Antiactin, Anti-His
antibodies were purchased from Santa Cruz Biotechnology (CA), The phosphatase inhibitor okadaic
acid (OA) was purchased from Sigma Life Sciences (Atlanta, GA). Endothall, a specific PP2A inhibitor,
was purchased from Calbiochem (La Jolla, CA). ATRA was obtained from Hoffman La Roche (Nutley,
NJ). Western blotting and immunoprecipitation, Phosphatase assay, Cloning and truncation,
Muta-genesis, Generation of CAOV3 cells were performed as described by Soprano DR et all. 2006).
Results: ATRA treatment leads to hypophosphorylation of PP2Ac, which correlates with increased
PP2A, a serine/threonine phosphate, and binds and dephosphorylates Rb2/p130 in the cell. We observed
that the N-terminus of PP2A catalytic subunit (PP2Ac) interacts with nuclear localization signals (NLS)
in the C-terminus of Rb2/p130. PP2Ac methylation is also increased following ATRA treatment and
PP2A methylating-enzyme (PMT) transcript levels are elevated as early as 6 hrs following ATRA
treatment. Immunoprecipitation studies reveal that PP2A and Rb2/p130 interact with importin β and
importin α, members of the nuclear transport machinery. Importin α recognizes a nuclear localization
signal (NLS) on a target protein and importin β binds the nuclear pore complex. CAOV3 cells
transfected with a truncated C-terminus region of Rb2/p130 in which the NLSs were mutated, no
interaction with PP2A was observed and these cells were as sensitive to ATRA treatment as parental
CAOV3 cells. This suggests that Rb2/p130 interaction with PP2A is required for the induction of cell
cycle arrest in ATRA treated cells. Conclusions: The results suggest that ATRA treatment leads to a
variety of responses that promote cell cycle arrest in ATRA sensitive COAV3 cells, including the
activation of a key phosphatase, PP2A, which enhances the stability and nuclear location of a tumor
suppressor protein, Rb2/p130.
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No. 91 (PO 5)
Genome-wide association analysis with selective genotyping identifies candidate loci
for adult height at 8q21.13 and 15q22.33-q23 in Mongolians.
M.Enkhdelger 1,3 , T.Kimura 2 , T.Kobayashi 2 , B.Munkhbat 1,2 , G. Oyungerel 1 , H. Hayashi 2 A.Oka 2 , I.
Inoue 2 , H.Inoko 2 , N.Munkhtuvshin 1
1
Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
2
Department of Molecular Life Science, Tokai University School of Medicine, Isehara, Kanagawa,
Japan
3 CEO Laboratory, Monolab Co., Ulaanbaatar Mongolia
Material and methods: A total of 1,555 unrelated individuals of Khalkh-Mongolian origin from the
region of Ulaanbaatar, Mongolia participated in the study. Four DNA pools were prepared. The first set
was DNA pools of 125 male-tall, 125 male-short, 125 female-tall, and 125 female-short samples. A
second set was also grouped from another 125 male- and female-tall samples and 125 male- and
female-short samples. All microsatellite markers and methods for microsatellite genotyping used in this
study are described by Tamiya et al. (2005).
Results: We performed a genome-wide association study with 23,465 microsatellite markers for
detection of loci controlling adult height using the selective genotyping method. To reduce cost and
technical burden of genome-wide genotyping, the pooled DNA method was applied. Association results
with the pooled DNA method and following re-genotyping of individual DNAs using the same set of
1,000 screened individuals, 23 markers showed significant differences by Fisher’s exact test. These
markers were subjected to correction of multiple tests with the number of alleles, and nine
microsatellites remained significant. We detected significant association in chromosomes 3, 4, 8, 15,
and 17. In addition, five regions overlapped at least partially with loci previously reported by linkage
analysis (5q31, 6q25, 8q21.3, 8q21.13, and 21q21.1). We also detected six strongly associated regions,
4q13.2, 4q31.3, 5q21.3, 7p21.1, 7q11.22, and 19q13.2, which have not been reported before. Among the
nine most associated markers, we selected two: D8S0285i and D15S988, the most strongly associated
microsatellite, located at 8q21.13, and D15S988 was flanked by a candidate gene, SMAD3, located at
15q22.33. 82 SNPs were surveyed and genotyped in a total of 1,555 samples (1,000 screened samples
and additional 555 samples).
Conclusions: We have identified two candidate loci for adult height at 8q21.13 and 15q22.33-q23 in
Mongolians. Analysis of the remaining seven highly associated microsatellite markers should lead to
identification of new causative genes underlying adult height variation.
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No. 92 (PO 6)
Molecular analysis of HLA polymorphism in ethnic minority of Khoton-Mongolians
P.Nyamragchaa 1,3 , B.Munkhbat 1,2 , T.Sato 2 , M.Hagihara 2 , K.Sato 2 , A.Kimura 2 , K.Tsuji 2 ,
N.Munkhtuvshin 1
1
Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
2
Department of Transplantation Immunology, Tokai University, Japan
3
Dept. Laboratory Medicine, General Hospital, Dundgobi, Mongolia
Objectives: To investigate the allelic frequencies of HLA class 1 and class II genes in ethnic minority
of Khoton-Mongolians. Materials and Methods: The population studied consisted of 85 randomly
selected healthy Khoton-Mongolians living in the area of Tarialan sum, of Uvs aimag, Mongolia.
Results were compared with control population consisting of 41 randomly chosen healthy unrelated
Khalkh-Mongolians from Ulaanbaatar, Mongolia. Allelic polymorphism in HLA-A, and HLA-B genes
were investigated by the PCR-SSOP method. HLA class II genotyping for DRB1, DRB3, DRB5, DQA1,
DQB1 and DPA1 genes were performed by the PCR-SSOP method. The PCR-RFLP method was used
for HLA-DPB1 genotyping. Results: Mongolia is a central Asian country with a population of 2.7
million. According to the ethnological studies, over 20 ethnic groups inhabit in the territory of Mongolia.
Among these, the Khalkh population is the largest majority (80% of total). In contrast, the Khoton is a
very small, isolated ethnic group residing in the Uvs aimag, in northwestern Mongolia. The Khoton
population is of special interest because there is no consensus regarding their origin. They are Muslim
people and have mixed very little with other ethnic groups. We investigated the polymorphism of 2
HLA class 1 and 7 HLA class II loci in Khoton-Mongolians in comparison with Khalkh-Mongolian
controls. Combined analysis of HLA class I and class II alleles showed that the two Mongolian
populations examined have significant differences in their distribution of HLA alleles; significantly
higher frequency of HLA-B38, DQA1*0201, DQB1*0201, DPB1*0401 and significantly lower
frequency of HLA-A2, A33, DRB1*1102, DQA1*0103, DQB1*0603, DPB1*0201 and DPB1*1701
were found in minor population of Khoton-Mongolians as compared to major Khalkh-Mongolians.
Several haplotypes were found to be unique to Khoton-Mongolians, such as HLA-A30-B58, A29-B35,
A24-B53, DRB1*0301-DQA1*0502- DQB1*0201-DPA1*0101-DPB1*0401, DRB1*0701-
DQA1*0201-DQB1*0201-DPA1*0202-DPB1*0501, DRB1*0401- DQA1*0301-DQB1*0301-
DPA1*0101-DPB1*0402, DRB1*1001-DQA1*0101-DQB1*0501- DPA1*0202-DPB1*0501 and
DRB1*0405-DQA1*0301-DQB1*0401-DPA1*0101-DPB1*0201.
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No. 93 (PO 7)
TNF-β gene-polymorphism in population and clinical studies
B.Ankhchimeg 1,3 , B.Munkhbat 1,2 , B.Gansuvd 1,2 , G.Oyungerel 1 , T.Shimura 2 N.Munkhtuvshin 1
1
Central Scientific Research Laboratory, Nat’l Institute of Medicine, Mongolia
2
Department of Transplantation Immunology, Tokai University, Japan
3
Dept. Laboratory Medicine, General Hospital, Arkhangai, Mongolia
Objectives: Genes of TNF-α (cachectin) and TNF-β (lymphotoxin) are located between HLA-B and
HLA class III, and no allelic polymorphism has been determined in the region of TNF-α loci, where two
alleles TNF-β1 and TNF-β2 has been described, that can inherits in three forms and no gene
frequencies and disease associations yet been fully studied.
Materials and methods: Ncol enzyme based RFLP allelic frequency of TNF-β (lymphotoxin) gene
analysis was performed in 126 healthy Mongolians (85 Khoton , and 41 Khalkh ethnic groups) and 135
Japanese lung cancer patients studied in comparison to 165 normal healthy control subjects.
Results: Gene frequency of 5.5/5.5-kb allele revealed in 12.2% of Khalkh population, and this is much
more frequent in comparison to Khoton population, in whom it was revealed only in 3.5%. But TNF-β
10.5/10.5-kb allele was determined in 49 cases (57.4%) in Khoton population, that was very high in
frequencies in comparison to both Khalkh and Japanese healthy group (34.1% and 41.1%),
(p=0.013440). In the lung cancer patients, the TNF-β 10.5/10.5-kb allele was found at a low frequency,
38.5%, compared to 53.3% in normal controls (chi 2 = 7.51, P = 0.011, corrected P = 0.033, relative risk
= 0.77). In the relationship between the histologic types and the TNF beta 10.5/10.5-kb allele showed
low frequencies: 38.5% in adenocarcinoma, 38.2% in squamous cell carcinoma, and 27.8% in small cell
carcinoma, although no statistical difference was shown. In relation to the postoperative survival period,
the TNF beta 10.5/10.5-kb allele was associated with prolonged survival.
CONCLUSIONS: The TNF beta 10.5/10.5-kb allele may be associated with resistance to lung cancer
and with a better prognosis.
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No. 94 (PO 8)
The genealogical study of Charcot-Marie-Tooth disease prevalence among some
Families in Bayan-Uul soum of Gobi-Altai province
Batchimeg B 1 , Bilegtsaikhan Ts 1 , Oyungerel G 1 , Tselmen D 1 , Erdenechimeg Ya 2 , Oyuntsetseg М 3 ,
Baasanjav D 2 , Munkhtuvshin N 1 , Munkhbat B 1
1Central Scientific Research Laboratory, Institute of Medical Sciences, Mongolia
2Department of Neurology, Institute of Medical Sciences, Mongolia
3Department of Neurology, Central Hospital of Gobi-Altai Province
Objectives: The purpose of the present study is to elucidate genealogical and clinical features of
hereditary neuropathy in the several large kindreds of Gobi-Altai province, Mongolia.
Materials and methods: In the present study, we had selected five kindreds originated from Bayan-Uul
sum, Gobi-Altai province. Each participant was enrolled for genealogical and neurological examinations
according to the prepared questionnaire. We also have collected biological samples for further genetic
study. Genomic DNA was isolated from biological samples, and quantitative analysis of DNA was
determined by spectrophotometer and picogreen assays.
Results: Genealogical analysis revealed that there was linkage between two kindreds in the total
families enrolled into study, whereas no association was revealed among the other pedigrees. As an
external appearance of susceptibility gene or genomic regions of the hereditary neuropathy, the clinical
signs were inheritant in every generation, and the inheritance was not dependent on sample of gender. In
neurological examination, start age of hereditary neuropathy onset was for the first decade of life in 4
patients, for the second decade of life in 5 patients, and for the other members start in the age of over
than 20 years. Common clinical features of hereditary neuropathy were characterized by hypomimicand
mask shape face, muscular atrophy of upper and lower limbs, and pes cavus. Five female patients
were revealed with similar gynecological problems, which was a very interesting case.
Conclusion:
1. The hereditary neuropathy that occurred in the large kindreds of Bayan-Uul sum, Gobi-Altai province,
and the type of inheritance could be categorized as autosomal dominant.
2. Onset of hereditary neuropathy disease started mostly in the second decade of life. Common clinical
features of hereditary neuropathy were characterized by hypomimic and mask shape face, muscular
atrophy of upper and lower limbs, and pes cavus. Apart from general clinical features, the specific
complicated signs were related to metabolic disorders and pregnancy.
- 147 -

No. 95 (PO 9)
Number and morphology of mesothelial cells in peritoneal dialysis effluent as a
potential predictive marker for advanced peritoneal dysfunction
Mayumi Idei 1 , Yoko Tabe 1 , Kazunori Miyake 1 , Chieko Hamada 2 , Hiroyuki Takemura 3 , Hiroaki
Io 2 , Kiyoshi Ishii 3 , Takashi Horii 3 , Yasuhiko Tomino 2 , Akimichi Ohsaka 3 , Takashi Miida 1
1, Department of Clinical Laboratory Medicine, Juntendo University School of Medicine, Tokyo, Japan
2, Division of Nephrology, Department of Internal Medicine, Juntendo University School of Medicine ,
Tokyo, Japan
3, Division of Clinical Laboratory, Juntendo University Hospital, Tokyo, Japan
Encapsulating peritoneal sclerosis (EPS) is a serious complication of peritoneal dialysis (PD). A risk
of EPS increases with a development of peritoneal dysfunction where increased peritoneal permeability
is associated with exfoliation and loss of peritoneal mesothelial cells.
In this study, we investigated whether the number and morphology of mesothelial cells in peritoneal
dialysis effluent (PDE) reflect a peritoneal function, and predict increased susceptibility to EPS.
Materials and Methods: PDE samples were obtained from 33 PD patients (26 men, 7 women, PD
duration; 29.1 ± 27.4 months) with peritoneal equilibration test (PET) performed. The number of
mesothelial cells was counted in 36 mm 2 area of slide preparation. According to the classification of cell
morphology based on the life-cycle of mesothelial cells, mesothelial cells were classified into quiescent,
activated, phagocytic, and degenerating cells with the digital microscopy system CellaVision DM96.
PET was used to categorize patients into four transporter groups; high (H), high average (HA), low
average (LA), and low (L).
Results: The number of mesothelial cells in PDE samples was significantly lower in H transporter
group than HA (p < 0.05). Mean numbers of mesothelial cells were 2.7 ± 0.6 for H transporter group
(n=3), 15.8 ± 11.4 for HA (n = 16), and 14.8 ± 21.7 for LA (n = 11), and 11.3 ± 12.9 for L (n=3),
respectively. In morphological analysis, whereas both quiescent- and activated- mesothelial cells were
present in HA, LA, and L transporter groups, in H transporter group, however, quiescent cell type was
predominantly observed.
Conclusions: Decreased number of mesothelial cells in PDE of a high transporter membrane state,
suggesting the loss of peritoneal mesothelial cells during chronic peritoneal injury, might be a
convenient and useful predictive marker for advanced peritoneal dysfunction.
- 148 -

No. 96 (P0 10)
Regulatory and pro-inflammatory properties of CD4+ T-cell subsets defined by
CD45RA, CCR7, CD27, and CD28 in patients with rheumatoid arthritis
Fumichika Matsuki*†, Jun Saegusa*†, Yoshiaki Miyamoto†, Kenta Misaki†, Shunichi Kumagai*§,
and Akio Morinobu†
Department of Evidence-based Laboratory Medicine, Kobe University Graduate School of Medicine,
Kobe, Japan; †Department of Rheumatology and Clinical Immunology, Kobe University Graduate
School of Medicine, Kobe, Japan; §The Center for Rheumatic Diseases, Shinko Hospital, Kobe, Japan
Background/Purpose: The phenotypic classification of T cells is useful in studies of immunological
diseases. Human CD4+ T cells can be classified as either naive, central memory (TCM), or effector
memory (TEM) cells using CCR7 and CD45RA. We recently discriminated CD4+ T cells into five major
subsets using four markers, CD45RA, CCR7, CD27, and CD28, and defined the function of each
population based on its ability to produce IFN-γ, IL-4, and IL-2. To identify the CD4+ T cell subsets
most important in the pathogenesis of rheumatoid arthritis (RA), we phenotypically defined human
CD4+ T cells as functionally distinct subsets, and analyzed the distribution and characteristics of each
subset in the peripheral blood.
Methods: Peripheral blood mononuclear cells from RA patients and healthy subjects were classified
into different subsets based on the expression of CD45RA, CCR7, CD27, and CD28 using five-color
flow cytometry. The frequency of IFN-γ-, IL-17-, or TNF-α-producing cells, and of Foxp3- or
RANKL-positive cells in each subset was analyzed by eight-color flow cytometry.
Results:
We classified human CD4+ T cells into six novel subsets based on four cell surface markers, CD45RA,
CCR7, CD27, and CD28. We demonstrated that the CD27+CD28+ TCM subset comprised a significantly
smaller proportion of CD4+ T cells in RA patients compared to healthy controls, and within this subset,
the proportion of Foxp3-positive cells was lower. In contrast, the proportion of IL-17- and
TNF-α-producing cells in the CD27+CD28+ TEM subset was significantly increased in RA patients.
Conclusion:
These findings suggest that the total number and/or proportion of inflammatory cytokine- or
Foxp3-positive cells of particular CD4+ T cells subsets is significantly changed in RA patients. These
subsets may provide novel therapeutic targets for this disease.
- 149 -

No. 97 (PO 11)
5-Ethynyl-2′-deoxyuridine (EdU)-induced replication banding patterns
in human chromosomes
Osamu Hoshi
Anatomy and Physiological Science, Graduate School of Health Care Science, Tokyo Medical and
Dental University, Japan
We present a novel technique using incorporation of 5-ethynyl-2′-deoxyuridine (EdU) into replicating
DNA is reported for analysis of the replicating banding patterns of human metaphase chromosomes.
Human lymphocytes were synchronized with excess thymidine, and then treated with EdU during the
late S phase of the cell cycle. EdU-incorporated DNA was detected in metaphase chromosomes using
Alexa Fluor ® 488 azides, through the 1,3-dipolar cycloaddition reaction of organic azides with the
terminal acetylene group of EdU. Chromosomes with incorporated EdU during the late S phase showed
a banding pattern that was similar to the G-banding pattern of normal human chromosomes. Atomic
force microscopic (AFM) images under liquid conditions showed that the structure of the chromosomes
was well preserved even after EdU treatment. Comparisons between fluorescence microscopic images
and AFM images of the same chromosome 1 indicated the presence of ridges and grooves in the
chromatid arm, which corresponded to G-positive and G-negative bands, respectively. These results
suggest an intimate relationship between the EdU-induced replication bands and the G-bands in human
chromosomes. This technique combined with AFM is useful for analyzing the structure of chromosomes
in relation to their replication banding pattern.
- 150 -

No. 98 (PO 12)
Chaotic Nature of Myocardial Ultrasonic Radio Frequency Signals Reflect Cardiac
Fibrosis and Hypertrophy in Patients with Hypertension.
Mitsuru Masaki, M.D *# ; Akiko Goda, M.D * ; Masataka Sugahara, M.D * ; Miho Fukui, M.D *# ;
Minoru Murakami, M.T # ; Miho Kuroda, M.T # ; Ayumi Igaki, M.T # ; Seiji Fujii, M.T # ; Tadanao
Wada, M.T # ; Kohji Inuzumi, M.T # , Masahiro Koshiba, M.D # ; and Tohru Masuyama, M.D *
Cardiovascular Division, Department of Internal Medicine, Hyogo College of Medicine *
Division of Clinical Laboratory Medicine, Hyogo College of Medicine #
Background: Chaotic nature of myocardial ultrasonic radio-frequency (RF) signals have been reflected
myocardial tissue properties. However, it is unclear whether the ultrasonic RF signals reflect cardiac
fibrosis in patients with hypertension. Methods: We evaluated 33 patients with untreated hypertension,
randomly assigned to calcium-channel blockers or Angiotensin II receptor blockers for 12 months. RF
signals were obtained from the myocardium by the transthoracic echocardiography. We assessed the
correlation dimension (CD) in left ventricular posterior wall at end diastole before and after treatment.
Plasma adiponectin, matrix metalloproteinase type 1(MMP-1), tissue inhibitor of metalloproteinase type
1 (TIMP-1) and aldosterone were determined with ELISA as cardiac fibrotic markers before and after
treatment. Results: CD, serum adiponectin, MMP-1 and TIMP-1 did not change after treatment. While,
serum aldosterone decreased after treatment (10.0 (5.5) vs. 13.3 (6.3) ng/dl, p

No. 99 (PO 13)
Drug susceptibility test for Giardia intestinalis using WST-1 reagent
Takahiro Matsumura *,** , Tomoji Yuno * , Masaharu Tokoro **
* Department of Clinical Laboratory, Kanazawa Red Cross Hospital
** Department of Parasitology, Graduate School of Medical Science, Kanazawa University, Japan
In human giardiasis, remaining diarrhea after chemotherapy is not rare, due to low compliance with the
therapy, repeated infections or parasite resistance to administrated drugs. Actually, in some clinical
reports, the emergence of drug resistance have been suggested as a cause of treatment failure in
giardiasis, however, the method for the drug susceptibility test of Giardia intestinalis and the higher
success rate of isolation method is not yet optimized. In contrast to the field of bacteriology, such as
methicillin-resistant Staphylococcus aureus (MRSA) and penicillin-resistant Streptococcus pneumoniae
(PRSP) etc. Considering these difficult conditions of giardiasis, we have taken a challenge to establish
the metronidazole susceptibility test for G. intestinalis. Culture isolation to the axenic condition using
modified YI-S medium (consisting of a nutrient broth, vitamin mixture and serum) of G. intestinalis
was conducted using cysts excreted in patient feces, and 4 out of 6 strains were successfully isolated.
The determination of Giardia genotype was used with molecular biological analysis. Using the
well-grown clinical 3 strains (Su2: assemblage B, Su3: assemblage A, Su4: assemblage A) along with
reference strain Portland-I (PO-1: assemblage A), the metronidazole susceptibility was assessed with a
bioluminescent WST-1 cell proliferation assay (Takara: Japan). The IC50 values of PO-1 and clinical 3
isolates (Su2, Su3, Su4) were 9.36 and 9.45, 6.58, 11.72 (μM) respectively for metronidazole. Our
method could isolate both assemblage A and B, common genotypes detection from humans, and the
WST-1 reagent could confirm the drug susceptibility in the culture condition. However, some genotypes
of Giadia have been considered to be difficult to adopt axenic culture condition. So, the best culture
conditions hold the key to the assessment of the drug susceptibility for Giardia. In fact, effective drug
choices according to the results of drug susceptibility tests are general protocol in bacterial diseases, and
such a useful assessment method is now available even for giardiasis. This study might help to explain
the diversity in drug susceptibility for Giardia.
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[Memo]
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