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Sleep Analysis Software Sleep. Sign® for Animal Sleep Analysis Software Sleep. Sign® for Animal

Summary 1. History Industry standard software developed with OBI and accepted world wide 2. Summary 1. History Industry standard software developed with OBI and accepted world wide 2. Data acquisition Any devices can be integrated 3. Sleep stage scoring Easy enough for beginners Sophisticated enough for experts 4. Post analysis Quick and easy reporting

Osaka Bioscience Institute • Discovered somnogenic effect of PGD 2 • Citation of published Osaka Bioscience Institute • Discovered somnogenic effect of PGD 2 • Citation of published scientific papers, world NO. 1 • KISSEI co-developed Sleep. Sign Osaka Japan http: //www. obi. or. jp

International customers • 13 countries, 240 licenses • Cited by most highly ranked scientific International customers • 13 countries, 240 licenses • Cited by most highly ranked scientific papers Stanford University Center for Narcolepsy Harvard Medical School Department of Neurology Japan Osaka Bioscience Institute Molecular Behavioral Biology USA Eastern Virginia Medical College Tokyo Medical and Dental University Department of Pathology and Anatomy Institute of Biomaterials and Bioengineering

Beijing university Korea Institute of Science and Technology Center for Neuroscience Department of pharmacology Beijing university Korea Institute of Science and Technology Center for Neuroscience Department of pharmacology Eli Lilly Pfizer Asia India Jawahar Lal Nehru University Fudan university Chinese university of Hong Kong EU Center for Neuroscience Waseda-Olympus Institute. Singapore Center for Neuroscience Hôpital Pellegrin. Boudreaux France Glaxo. Smith. Kline

Digital data import BIOPAC Sleep. Sign Import directly DSI Dataquest A. R. T EDF Digital data import BIOPAC Sleep. Sign Import directly DSI Dataquest A. R. T EDF file (European Data Format)

Analogue recording A/D conversion card Analogue amplifier Analogue recording A/D conversion card Analogue amplifier

Analogue recording + Video Camera MPEG encoder card A/D conversion card Analogue amplifier Analogue recording + Video Camera MPEG encoder card A/D conversion card Analogue amplifier

EEG is a wave A wave is defined by Amplitude and Frequency Amplitude Frequency EEG is a wave A wave is defined by Amplitude and Frequency Amplitude Frequency

Human sleep EEG Human sleep EEG

FFT (Fast Fourier Transformation) FFT maps the power of each frequency bins in a FFT (Fast Fourier Transformation) FFT maps the power of each frequency bins in a wave Waveform Power spectra 1 sec μV 2 2 Hz Hz μV 2 Hz

Rat sleep stages Rat sleep stages

Wake to NREM Wake NREM EEG FFT High amplitude slow wave EMG becomes flat Wake to NREM Wake NREM EEG FFT High amplitude slow wave EMG becomes flat

NREM to REM NREM REM EEG FFT Low amplitude, theta wave EMG is flat NREM to REM NREM REM EEG FFT Low amplitude, theta wave EMG is flat

REM to Wake REM Wake EEG Not rhythmical FFT EMG becomes active REM to Wake REM Wake EEG Not rhythmical FFT EMG becomes active

Automatic scoring • Easy operation • Computer assists criteria decision • 85 -95% agreement Automatic scoring • Easy operation • Computer assists criteria decision • 85 -95% agreement to human visual scoring

Various analysis parameters The number of brief wake Delta power for sleep stage Total Various analysis parameters The number of brief wake Delta power for sleep stage Total amount of each stage Duration of sleep wake episodes Sleep onset Quality of sleep Quantity of sleep Evaluate efficacies on sleep regulation

The number of Brief wake (Hypnogram) Timescale graph of sleep stages The number of Brief wake (Hypnogram) Timescale graph of sleep stages

Brief Wake between NREM Sleep in Mice brief wake (<16 s) NREM EEG NREM Brief Wake between NREM Sleep in Mice brief wake (<16 s) NREM EEG NREM EMG 4 s W WT R N W H 1 R KO R N 8: 00 8: 15 Huang, et al. , PNAS 2006 8: 30 8: 45

Total amount of sleep stage (Stage Graph) Total amount spent in each sleep stages Total amount of sleep stage (Stage Graph) Total amount spent in each sleep stages per time bins Non. REM Wake REM Light Dark

Mizoguchi & Eguchi et al. , PNAS 2001 Mizoguchi & Eguchi et al. , PNAS 2001

Duration of sleep episodes (Vigilance State Parameters) Duration of sleep episodes (Vigilance State Parameters)

Huang, et al. , PNAS 2006 Huang, et al. , PNAS 2006

The amount of delta power (Average FFT) Stage dependent average FFT per time bin The amount of delta power (Average FFT) Stage dependent average FFT per time bin NREM/Light’ ’’’’’’’' NREM/Dark

Multi Animal Screening New function Multi Animal Screening New function

Rat sleep stages Rat sleep stages

Simple interface SD Rat_4 SD Rat_3 SD Rat_2 SD Rat_1 multi screening is executed Simple interface SD Rat_4 SD Rat_3 SD Rat_2 SD Rat_1 multi screening is executed after pre-setting common value of screening parameters Simple pre –setting parameter menu

Initialize Scoring Ratio Step 1: calc each portions of R/W/NR R 8% W 12% Initialize Scoring Ratio Step 1: calc each portions of R/W/NR R 8% W 12% NR 80% Step 2: multiple ratio of Light/Dark phase NR 80% Dark phase 20% NR 16% Dark phase 20% R 1. 6% W 2. 4% NR 16% X Light phase 80% NR 64% Light phase 80% R 6. 4% W 9. 6% NR 64%

Adjusting rat Dark phase EMG integral (m. V/sec) EEG q/(d+q) ratio(%) EEG d power Adjusting rat Dark phase EMG integral (m. V/sec) EEG q/(d+q) ratio(%) EEG d power (m. V 2) Locomotio n activity (count) Light phase W R NR 12: 00 14: 00 16: 00 24: 00 Clock time 02: 00 04: 00

Locmoter Locmoter

Summary 1. History Industry standard software developed with OBI and accepted world wide 2. Summary 1. History Industry standard software developed with OBI and accepted world wide 2. Data acquisition Any devices can be integrated 3. Sleep stage scoring Easy enough for beginners Sophisticated enough for experts 4. Post analysis Quick and easy reporting

Acknowledgments Dr. T. Mochizuki (Beth Israel Deaconess Medical Center and Harvard Medical School) Dr. Acknowledgments Dr. T. Mochizuki (Beth Israel Deaconess Medical Center and Harvard Medical School) Dr. X. Tang (Eastern Virginia Medical School) Dr. Z. Huang (Fudan University and Osaka Bioscience Institute) Dr. O. Hayaishi, Dr. Y. Urade (Osaka Bioscience Institute) Dr. S. Nishino, N. Fujiki (Stanford University, Center for Narcolepsy) Dr. Z. Hu (Third Military Medical University) Dr. S. Inoue, K. Honda (Tokyo Medical and Dental University) Dr. N. Eguchi (Waseda-Olympus Bioscience Reserach Institute)