Chemical, Gas, and Biosensors for

Internet of Things and Related

Applications Kohji Mitsubayashi
Visit to download the full and correct content document:
https://ebookmass.com/product/chemical-gas-and-biosensors-for-internet-of-things-a
nd-related-applications-kohji-mitsubayashi/
Chemical, Gas, and Biosensors for
Internet of Things and Related
Applications
This page intentionally left blank
Chemical, Gas, and
Biosensors for Internet of
Things and Related
Applications

Edited by

Kohji Mitsubayashi
Department of Biomedical Devices and
Instrumentation, Institute of Biomaterials and
Bioengineering, Tokyo Medical and Dental
University, Tokyo, Japan

Osamu Niwa
Saitama Institute of Technology, Fukaya, Japan

Yuko Ueno
NTT Basic Research Laboratories,
NTT Corporation, Atsugi, Japan
Elsevier
Radarweg 29, PO Box 211, 1000 AE Amsterdam, Netherlands
The Boulevard, Langford Lane, Kidlington, Oxford OX5 1GB, United Kingdom
50 Hampshire Street, 5th Floor, Cambridge, MA 02139, United States
Copyright © 2019 Elsevier Inc. All rights reserved.
No part of this publication may be reproduced or transmitted in any form or by any means, electronic or
mechanical, including photocopying, recording, or any information storage and retrieval system, without
permission in writing from the publisher. Details on how to seek permission, further information about
the Publisher’s permissions policies and our arrangements with organizations such as the Copyright
Clearance Center and the Copyright Licensing Agency, can be found at our website: www.elsevier.com/
permissions.
This book and the individual contributions contained in it are protected under copyright by the Publisher
(other than as may be noted herein).
Notices
Knowledge and best practice in this field are constantly changing. As new research and experience
broaden our understanding, changes in research methods, professional practices, or medical treatment
may become necessary.
Practitioners and researchers must always rely on their own experience and knowledge in evaluating and
using any information, methods, compounds, or experiments described herein. In using such
information or methods they should be mindful of their own safety and the safety of others, including
parties for whom they have a professional responsibility.
To the fullest extent of the law, neither the Publisher nor the authors, contributors, or editors, assume
any liability for any injury and/or damage to persons or property as a matter of products liability,
negligence or otherwise, or from any use or operation of any methods, products, instructions, or ideas
contained in the material herein.
British Library Cataloguing-in-Publication Data
A catalogue record for this book is available from the British Library
Library of Congress Cataloging-in-Publication Data
A catalog record for this book is available from the Library of Congress
ISBN: 978-0-12-815409-0

For Information on all Elsevier publications

visit our website at https://www.elsevier.com/books-and-journals

Publisher: Susan Dennis

Acquisition Editor: Kathryn Morrissey
Editorial Project Manager: Vincent Gabriel
Production Project Manager: Prem Kumar Kaliamoorthi
Cover Designer: Greg Harris
Typeset by MPS Limited, Chennai, India
Contents

List of Contributors xv
Preface xix

Part I Sensors and Devices for Internet of Things

Applications 1
1 Portable urine glucose sensor 3
Narushi Ito, Mariko Miyashita and Satoshi Ikeda
1.1 Introduction 3
1.2 Significance of urine glucose measurement 3
1.3 Operating principle of urine glucose sensor and laminated structure 4
1.3.1 Principle of operation 4
1.3.2 Laminated structure of urine glucose sensor 5
1.4 Development of portable urine glucose meter 6
1.4.1 Composition of urine glucose meter 6
1.4.2 Performance evaluation of urine glucose meter 7
1.5 Clinical application of urine glucose meter 8
1.5.1 Relationship between the amount of boiled rice and
urine glucose concentration in impaired glucose tolerance 8
1.5.2 Results of urine glucose monitoring on impaired glucose
tolerance case 8
1.5.3 Results of a case of self-monitoring of urine glucose in
diabetes 10
1.6 Conclusions 11
References 11

2 Design, application, and integration of paper-based sensors

with the Internet of Things 13
Jen-Hsuan Hsiao, Yu-Ting Tsao, Chung-Yao Yang and Chao-Min Cheng
2.1 Introduction 13
2.2 Bioapplications of paper-based analytical devices 14
2.3 Environmental analysis of paper-based analytical devices 17
2.4 Integration with smartphone devices 19
2.5 Conclusion 24
Author disclosure statement 24
References 24
vi Contents

3 Membrane-type Surface stress Sensor (MSS) for artificial olfactory

system 27
Huynh Thien Ngo, Kosuke Minami, Gaku Imamura, Kota Shiba
and Genki Yoshikawa
3.1 Introduction 27
3.2 Membrane-type Surface stress Sensor (MSS) 28
3.3 Receptor materials 29
3.4 Machine learning 33
3.5 Applications 34
3.6 Internet of Things and MSS Alliance/Forum 36
3.7 Conclusion 37
References 37

4 Sensing technology based on olfactory receptors 39

Hidefumi Mitsuno, Takeshi Sakurai and Ryohei Kanzaki
4.1 Olfactory mechanisms in biological systems 39
4.1.1 Olfactory mechanisms in vertebrates 39
4.1.2 Olfactory mechanisms in insects 41
4.2 Biosensing technologies based on odorant receptors 45
4.2.1 Mammalian odorant receptors 47
4.2.2 Insect odorant receptors 53
4.3 Summary 58
References 59

5 Advanced surface modification technologies for biosensors 65

Chun-Jen Huang
5.1 Biosensors and biointerfaces 65
5.2 Binding platforms based on self-assembled monolayers 66
5.2.1 Organosulfur derivatives 68
5.2.2 Organosilicon derivatives 69
5.2.3 Catechol derivatives 70
5.3 Binding matrix based on polymeric hydrogels 71
5.3.1 Physicochemical sensing mechanisms 71
5.3.2 Biochemical sensing mechanisms 72
5.4 Coupling chemistries for immobilization of biorecognition elements 73
5.4.1 Physical immobilization 73
5.4.2 Amine chemistry 73
5.4.3 Thiol chemistry 74
5.4.4 Carboxyl chemistry 75
5.4.5 Epoxy chemistry 76
5.4.6 Click chemistry 76
5.4.7 α-Oxo semicarbazone chemistry 77
5.4.8 Bioaffinity conjugation 77
Contents vii

5.5 Antifouling materials 78

5.5.1 Poly(ethylene glycol) antifouling materials 78
5.5.2 Zwitterionic antifouling materials 78
5.6 Outlook 79
References 79

6 Development of portable immunoassay device for future Internet

of Things applications 87
Ryoji Kurita, Osamu Niwa and Yuko Ueno
6.1 Introduction 87
6.2 Portable immunoassay system based on surface plasmon
resonance for urinary immunoassay 88
6.3 One-chip immunosensing fabricated with nanoimprinting
technique 92
6.3.1 Fabrication of local plasmon resonance devices with
various processes 92
6.3.2 Surface plasmon resonance biosensors fabricated by
nanoimprint technique 94
6.4 Microfluidic biosensor with one-step optical detection 95
6.4.1 Mechanism of graphene aptasensor 97
6.4.2 Multichannel linear array for multiple protein detection 98
6.4.3 Molecular design for enhanced sensitivity 99
6.5 Future trend 100
References 101

7 Sensitive and reusable surface acoustic wave immunosensor for

monitoring of airborne mite allergens 105
Koji Toma, Takahiro Arakawa and Kohji Mitsubayashi
7.1 Introduction 105
7.2 Surface acoustic wave immunosensor for repeated measurement
of house dust mite allergens 106
7.3 Sensor characteristics and semicontinuous measurement of
Der f 1 109
7.4 Sensitivity improvement via gold nanoparticles 111
7.5 Conclusion 115
References 115

8 Aptameric sensors utilizing its property as DNA 117

Kinuko Ueno, Kaori Tsukakoshi and Kazunori Ikebukuro
8.1 Introduction 117
8.2 Aptamer-immobilized electrochemical sensor 117
8.3 Detection using complementary chain formation 118
8.3.1 Strand displacement assay 119
viii Contents

8.3.2 Bound/Free separation using complementary

chain formation 119
8.4 Aptamer sensor combined with enzymes 121
8.5 Utilizing structural change of aptamers to biosensor 121
8.6 Utilizing structural change of aptamers to biosensor 122
8.7 Development of highly sensitive sensors by amplifying
DNA strands 125
8.8 Colorimetric detection using aptameric sensor and
smart devices 126
8.9 Conclusion 128
References 128

9 Electrochemical sensing techniques using carbon electrodes

prepared by electrolysis toward environmental Internet of
Things sensor 133
Hiroaki Matsuura
9.1 Introduction 133
9.1.1 Electrochemical monitoring support Internet of
Things services 133
9.1.2 Carbon electrode surface activation 135
9.2 Chemical sensors using electrochemical activated carbon
electrodes 136
9.2.1 Electrochemical activated techniques for aminated
electrode preparation 136
9.2.2 Electrochemical activated techniques for
electrodeposited platinum particles on glassy carbon
electrode modified with nitrogen-containing functional
groups 136
9.3 Electrocatalytic activity and analytical performance 136
9.4 Conclusion and future perspectives 143
Acknowledgments 143
References 144

10 Chemical sensors for environmental pollutant determination 147

Hongmei Bi and Xiaojun Han
10.1 Introduction 147
10.2 Definition of a chemical sensor 147
10.3 Classification of chemical sensors 148
10.3.1 Electrochemical sensors 148
10.3.2 Optical sensors 152
10.4 Conclusion 158
Acknowledgments 158
References 158
Contents ix

Part II Flexible, Wearable, and Mobile Sensors

and Related Technologies 161
11 Smart clothing with wearable bioelectrodes “hitoe” 163
Hiroshi Nakashima and Shingo Tsukada
11.1 Introduction 163
11.2 Functional material “hitoe” 165
11.2.1 Composite material of a conductive polymer and fibers 165
11.2.2 The development of hitoe smart clothing 166
11.3 Application examples 168
11.3.1 Medicine/rehabilitation 168
11.3.2 Sports 169
11.3.3 Worker health/safety management 172
11.4 State estimation based on heart rate variability and other data 173
11.4.1 Estimating posture information from accelerometer data 173
11.4.2 Estimating respiratory activity from
electrocardiogram data 174
11.4.3 Estimating sleep states 174
11.5 Conclusion 175
References 176

12 Cavitas bio/chemical sensors for Internet of Things in healthcare 177

Kohji Mitsubayashi, Koji Toma and Takahiro Arakawa
12.1 Introduction 177
12.2 Soft contact lens type bio/chemical sensors 179
12.2.1 Tear fluid in conjunctiva sac 179
12.2.2 Flexible conductivity sensor for tear flow function 179
12.2.3 Soft contact lens type biosensors using biocompatible
polymers 181
12.2.4 Transcutaneous gas sensor at eyelid conjunctiva 183
12.3 Mouthguard type biosensor for saliva biomonitoring 185
12.3.1 Salivary fluids in oral cavity 185
12.3.2 Wireless mouthguard sensor for salivary glucose 185
12.4 Conclusion 188
Acknowledgments 188
References 189

13 Point of care testing apparatus for immunosensing 193

Tomoyuki Yasukawa, Fumio Mizutani and Masato Suzuki
13.1 Introduction 193
13.2 Immunochromatography assay 195
13.3 Immunochromatography assay for infectious diseases 197
13.4 Reliability of the examination kits 198
x Contents

13.5 Signal amplification 198

13.6 Quantitative ICA by electrochemical detection systems 199
13.7 Rapid and Quantitative ICA based on dielectrophoresis 200
13.8 Conclusion 202
References 203

14 IoT sensors for smart livestock management 207

Wataru Iwasaki, Nobutomo Morita and Maria Portia Briones Nagata
14.1 Introduction 207
14.2 Measurement site and fixing method 209
14.3 Size and weight 209
14.4 Power consumption 210
14.5 Frequency bands of radio wave 210
14.6 Applications of wearable biosensors for livestock 211
14.6.1 Chickens 211
14.6.2 Cattle 213
14.7 Conclusion 218
References 218

15 Compact disc-type biosensor devices and their applications 223

Izumi Kubo and Shunsuke Furutani
15.1 Introduction 223
15.2 CD-shaped microfluidic devices for cell isolation and
single cell PCR 224
15.2.1 Single cell isolation 224
15.2.2 Single cell PCR of S. enterica 225
15.2.3 Discrimination of microbes 226
15.2.4 Single cell RT-PCR for Jurkat cells 227
15.3 CD-shaped microfluidic device for cell staining 228
15.4 CD-shaped microfluidic device for ELISA 230
15.4.1 Detection of bioactive chemicals based on ELISA 230
15.4.2 Multiple ELISA for diagnosis of diabetes 232
15.5 Conclusion 233
Acknowledgment 234
References 234

16 A CMOS compatible miniature gas sensing system 237

Ting-I Chou, Shih-Wen Chiu and Kea-Tiong Tang
16.1 Introduction 237
16.2 Complementary metal
oxide
semiconductor-compatible
gas sensor 238
16.2.1 Materials and fabrication 238
16.2.2 Gas experimental results 239
Contents xi

16.3 Nose-on-a-chip 241

16.3.1 System block diagram 241
16.3.2 Adaptive interface circuitry 241
16.3.3 SAR ADC 242
16.3.4 CRBM kernel 243
16.3.5 Memory 244
16.3.6 RISC core 245
16.3.7 Chip measurement results 245
16.4 Miniature electronic nose system prototype 247
16.5 Application example 248
16.6 Conclusion 250
Acknowledgments 251
References 251

17 Visualization of odor space and quality 253

Fumihiro Sassa, Chuanjun Liu and Kenshi Hayashi
17.1 Introduction 253
17.2 Fluorescence imaging for odor visualization 255
17.2.1 Principle and system of fluorescence imaging 255
17.2.2 Fabrication of the visualization system 256
17.2.3 Visualization based on single fluorescent probe 256
17.2.4 Visualization based on multispectral fluorescence
imaging 258
17.3 Localized surface plasmon resonance sensor for odorant
visualization 260
17.4 Collecting spatial odor information from on-ground odor
sources with a robot system 261
17.5 Visual odor representation of a volatile molecular based
on chemical property by network diagram 264
References 266

18 Bio-sniffer and sniff-cam 271

Takahiro Arakawa, Koji Toma and Kohji Mitsubayashi
18.1 Introduction: breath and skin gas analysis 271
18.1.1 Construction of bio-sniffer 272
18.1.2 Acetone bio-sniffer 273
18.1.3 Isopropanol bio-sniffer 276
18.1.4 Sniff-cam system with chemiluminescence 277
18.1.5 Biofluorometric “sniff-cam” 281
18.2 Summary 285
Acknowledgments 285
References 285
xii Contents

Part III Information and Network Technologies for

Sensor-Internet of Things Applications 289
19 Flexible and printed biosensors based on organic TFT devices 291
Kuniaki Nagamine and Shizuo Tokito
19.1 Introduction 291
19.1.1 Biosensors for the Internet of Things society 291
19.1.2 Printed organic biosensors for human healthcare
applications 292
19.2 Organic thin-film transistor-based biosensors 293
19.2.1 Printing techniques for device fabrication 293
19.2.2 Organic thin-film transistor-based biosensor principles 293
19.2.3 Enzyme-based biosensors 295
19.2.4 Immunosensors 297
19.2.5 Ion-selective sensors 298
19.2.6 Wearable sensors using microfluidics 299
19.3 Sensor systems using flexible hybrid electronics 300
19.4 Conclusion 301
Acknowledgments 302
References 302

20 Self-monitoring of fat metabolism using portable/wearable

acetone analyzers 307
Satoshi Hiyama
20.1 Introduction 307
20.2 Portable breath acetone analyzer 307
20.2.1 Prototyped analyzer 308
20.2.2 Applicability to diet support 311
20.2.3 Applicability to diabetes care at home 312
20.2.4 Applicability to “Health Kiosk” 315
20.3 Wearable skin acetone analyzer 316
20.3.1 Skin acetone concentrator 316
20.3.2 Prototyped analyzer 318
20.3.3 Assumed usage scenario 318
20.4 Conclusions 320
References 320

21 Air pollution monitoring network of PM2.5, NO2 and

radiation of 137Cs 323
Yasuko Yamada Maruo
21.1 Introduction 323
21.2 PM2.5 monitoring system 324
21.2.1 Introduction 324
21.3 Monitoring device (small PM2.5 sensor) 324
Contents xiii

21.4 Mobile sensing of outside PM2.5 326

21.5 Measurement at several points 327
21.6 NO2 monitoring system 327
21.6.1 Introduction 327
21.7 NO2 monitoring device 329
21.8 Mobile sensing of outside NO2 331
21.9 Radiation of 137Cs monitoring system 331
21.9.1 Introduction 331
21.10 Radiation of 137Cs monitoring device 333
21.11 Field test in Fukushima and other areas 334
Acknowledgment 335
References 335

22 Wireless sensor network with various sensors 339

Junichi Kodate
22.1 Sensing system with network 339
22.2 Wireless sensor network as a sensing system 339
22.3 Wireless sensing system for health condition monitoring
with a wearable and flexible sensor 341
22.3.1 Wearable and flexible electrode with a conductive fiber 342
22.3.2 Wireless data-transmitting module with many sensors 342
References 343

23 Data analysis targeting healthcare-support applications using

Internet-of-Things sensors 345
Akihiro Chiba, Kana Eguchi and Hisashi Kurasawa
23.1 Motivation for data analysis 345
23.2 Procedure of data analysis 346
23.2.1 Analysis design 346
23.2.2 Data collection 349
23.2.3 Data cleansing 350
23.2.4 Feature extraction 351
23.2.5 Learning 354
23.2.6 Evaluation 355
23.3 Example of health data analysis 359
23.4 Conclusion 360
References 361

Summary and future issue 363

Index 365
This page intentionally left blank
List of Contributors

Takahiro Arakawa Department of Biomedical Devices and Instrumentation,

Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental
University, Tokyo, Japan

Hongmei Bi College of Science, Heilongjiang Bayi Agricultural University,

Daqing, P.R. China

Chao-Min Cheng Institute of Biomedical Engineering, National Tsing Hua

University, Hsinchu, Taiwan

Akihiro Chiba NTT Service Evolution Laboratories, Yokosuka, Japan

Shih-Wen Chiu National Tsing Hua University, Hsinchu, Taiwan

Ting-I Chou National Tsing Hua University, Hsinchu, Taiwan

Kana Eguchi NTT Service Evolution Laboratories, Yokosuka, Japan

Shunsuke Furutani Biomedical Research Institute, National Institute of Advanced

Industrial Science and Technology (AIST), Osaka, Japan; Advanced Photonics and
Biosensing Open Innovation Laboratory (PhotoBIO-OIL), National Institute of
AIST, Osaka, Japan

Xiaojun Han State Key Laboratory of Urban Water Resource and Environment,
School of Chemistry and Chemical Engineering, Harbin Institute of Technology,
Harbin, P.R. China

Kenshi Hayashi Department of Electronics, Graduate School of Information

Science and Electrical Engineering, Kyushu University, Fukuoka, Japan

Satoshi Hiyama Research Laboratories, NTT DOCOMO, Inc., Yokosuka, Japan

Jen-Hsuan Hsiao Department of Power Mechanical Engineering, National Tsing

Hua University, Hsinchu, Taiwan
xvi List of Contributors

Chun-Jen Huang Department of Biomedical Sciences and Engineering, National

Central University, Jhong-Li, Taiwan; Department of Chemical and Materials
Engineering, National Central University, Jhong-Li, Taiwan; R&D Center for
Membrane Technology, Chung Yuan Christian University, Chung-Li City, Taiwan

Kazunori Ikebukuro Department of Biotechnology and Life Science, Graduate

School of Engineering, Tokyo University of Agriculture and Technology, Tokyo,
Japan

Satoshi Ikeda TANITA Corporation, Tokyo, Japan

Gaku Imamura Center for Functional Sensor & Actuator (CFSN), National
Institute for Materials Science (NIMS), Tsukuba, Japan; International Research
Center for Materials Nanoarchitectonics (WPI-MANA), National Institute for
Materials Science (NIMS), Tsukuba, Japan

Narushi Ito PROVIGATE Inc., Tokyo, Japan

Wataru Iwasaki Advanced Manufacturing Research Institute, National Institute of

Advanced Industrial Science and Technology, Tokyo, Japan

Ryohei Kanzaki Research Center for Advanced Science and Technology, The
University of Tokyo, Tokyo, Japan

Junichi Kodate NTT Device Innovation Center, NTT Corporation, Atsugi, Japan

Izumi Kubo Graduate School of Bioinformatics, Soka University, Tokyo, Japan

Hisashi Kurasawa NTT Service Evolution Laboratories, Yokosuka, Japan

Ryoji Kurita National Institute of Advanced Industrial Science and Technology,

Tsukuba, Japan

Chuanjun Liu Research Laboratory, U.S.E. Co., Ltd, Tokyo, Japan

Yasuko Yamada Maruo Tohoku Institute of Technology, Sendai, Japan

Hiroaki Matsuura Department of Life Science & Green Chemistry, Faculty of

Engineering, Saitama Institute of Technology, Fukaya, Japan

Kosuke Minami Center for Functional Sensor & Actuator (CFSN), National
Institute for Materials Science (NIMS), Tsukuba, Japan; International Research
Center for Materials Nanoarchitectonics (WPI-MANA), National Institute for
Materials Science (NIMS), Tsukuba, Japan
List of Contributors xvii

Kohji Mitsubayashi Department of Biomedical Devices and Instrumentation,

Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental
University, Tokyo, Japan

Hidefumi Mitsuno Research Center for Advanced Science and Technology, The
University of Tokyo, Tokyo, Japan

Mariko Miyashita TANITA Corporation, Tokyo, Japan

Fumio Mizutani Graduate School of Material Science, University of Hyogo,

Kamigori, Japan

Nobutomo Morita Advanced Manufacturing Research Institute, National Institute

of Advanced Industrial Science and Technology, Tokyo, Japan

Kuniaki Nagamine Research Center for Organic Electronics (REOL), Yamagata

University, Yonezawa, Japan

Maria Portia Briones Nagata Advanced Manufacturing Research Institute,

National Institute of Advanced Industrial Science and Technology, Tokyo, Japan

Hiroshi Nakashima NTT Basic Research Laboratories, Nippon Telegraph and

Telephone Corporation, Tokyo, Japan

Huynh Thien Ngo Center for Functional Sensor & Actuator (CFSN), National
Institute for Materials Science (NIMS), Tsukuba, Japan; International Research
Center for Materials Nanoarchitectonics (WPI-MANA), National Institute for
Materials Science (NIMS), Tsukuba, Japan

Osamu Niwa Saitama Institute of Technology, Fukaya, Japan

Takeshi Sakurai Department of Agricultural Innovation for Sustainability, Faculty

of Agriculture, Tokyo University of Agriculture, Kanagawa, Japan

Fumihiro Sassa Department of Electronics, Graduate School of Information

Science and Electrical Engineering, Kyushu University, Fukuoka, Japan

Kota Shiba Center for Functional Sensor & Actuator (CFSN), National Institute
for Materials Science (NIMS), Tsukuba, Japan; International Research Center for
Materials Nanoarchitectonics (WPI-MANA), National Institute for Materials
Science (NIMS), Tsukuba, Japan

Masato Suzuki Graduate School of Material Science, University of Hyogo,

Kamigori, Japan
xviii List of Contributors

Kea-Tiong Tang National Tsing Hua University, Hsinchu, Taiwan

Shizuo Tokito Research Center for Organic Electronics (REOL), Yamagata

University, Yonezawa, Japan

Koji Toma Department of Biomedical Devices and Instrumentation, Institute of

Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo,
Japan

Yu-Ting Tsao School of Traditional Chinese Medicine, Chang Gung University

College of Medicine, Taoyuan, Taiwan

Shingo Tsukada NTT Basic Research Laboratories, Nippon Telegraph and

Telephone Corporation, Tokyo, Japan

Kaori Tsukakoshi Department of Biotechnology and Life Science, Graduate

School of Engineering, Tokyo University of Agriculture and Technology, Tokyo,
Japan

Kinuko Ueno Department of Biotechnology and Life Science, Graduate School of

Engineering, Tokyo University of Agriculture and Technology, Tokyo, Japan

Yuko Ueno NTT Basic Research Laboratories, NTT Corporation, Atsugi, Japan

Chung-Yao Yang Hygeia Touch Inc., Taipei, Taiwan

Tomoyuki Yasukawa Graduate School of Material Science, University of Hyogo,

Kamigori, Japan

Genki Yoshikawa Center for Functional Sensor & Actuator (CFSN), National
Institute for Materials Science (NIMS), Tsukuba, Japan; International Research
Center for Materials Nanoarchitectonics (WPI-MANA), National Institute for
Materials Science (NIMS), Tsukuba, Japan; Materials Science and Engineering,
Graduate School of Pure and Applied Science, University of Tsukuba, Tsukuba,
Japan
Preface

The growing interest in medical and environmental sensors can be attributed to the
increasing awareness that health is essential for enjoying and keeping a good qual-
ity of life. The advent of wearable and/or flexible sensors for healthcare represents
such a leap. The demand for quality healthcare has strained the current healthcare
system, which faces large workforce shortages and inadequate medical facilities. It
is paramount to develop consumer-based devices to augment the current healthcare
demands. One emerging technology is sensor networks connected to the Internet of
Things (IoT), a human-oriented network of tiny human monitoring devices devel-
oped on the body and in the surrounding infrastructure.
Chemicals, gas, and biosensors will be required for preventive medicine and
environmental assessment for healthcare demands in the near future. The reason
for this stems from the fact that all branches of modern medicine, ranging from
prevention to complex intervention, rely on early and accurate diagnosis followed
by close monitoring of the results. The research field of gas and bio/chemical sen-
sors has been developed by a group of truly interdisciplinary researchers involving
chemists, biologists, physicists, material scientists, and computer engineers to cre-
ate a range of novel configurations exploiting bio/chemical recognition systems
allied with physiochemical transducers. Novel chemical, gas, and biosensors
would further progress with the introduction of a range of optical, acoustic, mag-
netic, thermal, and electrical technologies, coupled with microelectronics and
MEMS devices for the future medical and healthcare systems. This book is orga-
nized with the following parts and chapters. Part 1 introduces novel bio/chemical
sensors with several recognition materials such as enzymes, antibodies, aptamers,
receptors, and artificial materials for analytes in body fluids and gas-phase sam-
ples. In Part 2, flexible and mobile sensors and related techniques are explained
with some wearable and cavitas (body cavity) sensors, and smell detectors.
Information and network technologies for the sensors are illustrated with industrial
applications in Part 3.
This book is intended for graduate students, academic researchers, and profes-
sors who work in the field of medical and environmental research, and also for
industry professionals involved in development of devices and systems with IoT
for human bio/chemical measurement, medical monitoring, and healthcare ser-
vices with Internet technologies. We would like to sincerely express our appre-
ciation to the distinguished authors of the chapters whose expertise has certainly
contributed significantly to the book. We hope that this book can shed light on
xx Preface

various technological aspects related to bio/chemical sensors with IoT and

related applications in a healthcare context, and will stimulate further research in
this field.

Kohji Mitsubayashi, Osamu Niwa and Yuko Ueno

Editors
Part I

Sensors and Devices for Internet

of Things Applications
This page intentionally left blank
Portable urine glucose sensor
1
Narushi Ito1, Mariko Miyashita2 and Satoshi Ikeda2
1
PROVIGATE Inc., Tokyo, Japan, 2TANITA Corporation, Tokyo, Japan

1.1 Introduction
Development of a noninvasive blood glucose monitoring system is based on mea-
surement of near infrared light passing through fingers and arms [1,2], measurement
of interstitial fluid collected from the skin surface with a micro glucose sensor
[3,4], and measurement of contact lens type tears [5,6], etc. It has been done over
the past 30 years and enormous amount of research and development has been pub-
lished, however, there are still no practical products.
A series of products that have succeeded in development include a continuous
blood glucose monitor and a flash glucose monitor that place small needles in the
abdomen and the like. These are minimally invasive, and continuous monitoring for
2 weeks is possible. Meanwhile, as a noninvasive measurement, a portable urine
glucose meter that makes it possible to quantitatively measure urine glucose levels
correlated with blood glucose levels has been put to practical use.
In this section, we describe the principle and structure of the microplanar type
urine glucose sensor and examples of application of commercialized urine glucose
meter to healthcare.

1.2 Significance of urine glucose measurement

In the urine glucose tests, diabetes screening tests are being conducted to test posi-
tive (1) or negative (2) by measuring fasting urine such as common in medical
examinations.
Positive (1) is based on 100 mg/dL as the urine glucose concentration.
Medically urine glucose positivity is considered as a suspicious indicator of diabe-
tes first. Then further inspections are necessary, because there are transient cases
such as renal diabetes, stress, pregnancy, etc., in addition to other findings for diag-
nosis of diabetes.
Ultimately diabetes is confirmed by the 75 g oral glucose tolerance test. Urine glu-
cose test is regarded as an auxiliary test and a large number of screening tests are still
being carried out at present, due to its advantage of noninvasive measurement.

Chemical, Gas, and Biosensors for the Internet of Things and Related Applications.
DOI: https://doi.org/10.1016/B978-0-12-815409-0.00001-2
© 2019 Elsevier Inc. All rights reserved.
4 Chemical, Gas, and Biosensors for the Internet of Things and Related Applications

Figure 1.1 Relationship between urine glucose concentration and blood glucose
concentration after meal.

Changes in urine glucose concentration after meals are shown in Fig. 1.1. The
postprandial urine glucose concentration is linked to blood glucose levels, and it is
important that it does not overlook postprandial hyperglycemia occurring after
meals. When the urine glucose concentration exceeds 50 mg/dL, it means that the
blood glucose level exceeds the glucose excretion threshold of 160
180 mg/dL in
the kidney.
Even when blood glucose level rises with the diet, it decreases after 1 hour due
to the action of insulin. In other words, the blood glucose level measured at 2 hours
after a meal usually returns to the normal range. On the other hand, it is known that
the urine glucose concentration 2 hours after a meal reflects the elevated blood glu-
cose level with the meal. As well, it is demonstrated that urine glucose level corre-
lates with the mean blood glucose level.

1.3 Operating principle of urine glucose sensor and

laminated structure
1.3.1 Principle of operation
Urine glucose sensor is an enzyme electrode method combining glucose oxidase
(GOX) and hydrogen peroxide electrode.
The electrode is fabricated by photolithography technology. Glucose is enzymat-
ically converted to hydrogen peroxide (H2O2) by GOX, and the yielded H2O2 is
electrochemically detected by the electrodes.
The enzymatic reaction (1) and the electrode reactions (2) are as follows:
1. Enzymatic reaction
GOX: Glucose 1 O2!gluconolactone 1 H2O2
2. Electrochemical reactions at electrodes
Working electrode: H2O2!2H1 1 O2 1 2e2
Portable urine glucose sensor 5

Figure 1.2 Perspective view of the sensor.

Counter electrode: 2H1 11/2O2 1 2e2!H2O

Entire electrode system: H2O2!H2O 1 1/2O2
A perspective view of the sensor is shown in Fig. 1.2. Three electrodes, a work-
ing electrode, a counter electrode, and a reference electrode, are formed in the hole
of the cartridge. The working electrode and the counter electrode are Pt electrodes,
and thin film Ag/AgCl electrodes are formed as reference electrode. The reference
electrode has the role of stabilizing the potential after immersion in the solution.
The outermost layer of the electrode is coated with a thin film of a fluorinated poly-
mer to prevent contamination due to urine components while protecting the elec-
trode system as a whole and stabilizing the operation of the electrodes for more
than 1 year in solution.

1.3.2 Laminated structure of urine glucose sensor

To accurately measure postprandial urine, means to eliminate the influence of vita-
min C (ascorbic acid) among substances released from foods are required. Ascorbic
acid has a reaction that gives electrons to an electrode and another reaction decom-
posing hydrogen peroxide, and it is typical of an interfering substance of an amper-
ometric sensor.
Furthermore, it is necessary to fabricate a membrane structure so that interfering
substances other than ascorbic acid contained in the urine do not affect the mea-
sured values.
Fig. 1.3 shows a laminated structure of the urine glucose sensor. This sensor is
composed of four layers: a restricted permeable layer, an enzyme immobilized
layer, a cation-exchanging layer, and an adhesive layer.
1. The restricted permeable layer has a wide measurement range from 10 to 2000 mg/dL,
limiting the diffusion of molecules larger than glucose. It has the role of preventing the
influence of adhered substances in urine.
2. The enzyme immobilized layer is the one where enzyme (GOX) and bovine serum albu-
min are crosslinked and immobilized so as not to inactivate the enzyme; as a result, repe-
tition of the sensor is possible.
6 Chemical, Gas, and Biosensors for the Internet of Things and Related Applications

Figure 1.3 Laminated structure of the urine glucose sensor.

3. The cation-exchanging layer has the role of permeating hydrogen peroxide and limiting
the diffusion of molecules larger than hydrogen peroxide. Furthermore, it has the function
of preventing permeation of ionized molecules.
4. The adhesive layer has the role of covalently bonding the selectively permeable film,
which is an organic material, to the surface of the glass substrate or the electrode and sta-
bly adhering for a long time in water.

This sensor is formed of a thin film of four layers with a total thickness of 1 μm
or less, effectively eliminating the influence of interfering substances in the urine
and an early time response [7].

1.4 Development of portable urine glucose meter

1.4.1 Composition of urine glucose meter
This urine glucose meter consists of a body and a sensor. Portability is designed so
that the sensor section is folded down to be compact, and at the time of measure-
ment it is extended. Photo 1.1 shows a urine glucose meter in a stored state. Photo
1.2 shows the urine glucose meter extended at the time of measurement. The urine
glucose meter sensor section is composed of a preservation solution bottle, which
makes the sensor wet. The preservation solution is reserved to hold the sensor,
which can cause optimal enzymatic reaction with pH buffer and physiological
saline. After the sensor is taken out, it becomes possible to measure instantly.
Photo 1.2 shows a urine glucose meter at the time of measurement. When mea-
sured, total length 210 mm of the meter can be directly applied to urine with one
hand. The urine glucose sensor at the tip is equipped with a thermistor for detecting
the water temperature so that the output can be corrected. The sensor needs to be
Portable urine glucose sensor 7

Photo 1.1 Urine glucose meter folded (closed).

Photo 1.2 Urine glucose meter extended (opened).

replaced either after 200 measurements or within 60 days due to the removable
socket structure. The main body measures the minute current and controls the
device. It also has a liquid crystal display that indicates urine glucose concentration,
and switches for calibration and measurement value recall. It also operates for 8
months with one lithium battery.

1.4.2 Performance evaluation of urine glucose meter

The final test of the urine glucose meter requires performance evaluation by the
urine of patients. The results of simultaneous measurement of patients’ urine speci-
mens with urine glucose meter and clinical urine glucose analyzer (A&T GA03R)
and correlation evaluation are shown in Fig. 1.4. The primary equation obtained by
the method of least squares is urine glucose meter ① Y 5 0.925 X 1 53.3,
R 5 0.987, urine glucose meter ② Y 5 0.939 X 1 62.9, R 5 0.987, urine glucose
meter ③ Y 5 0.968 X 1 40.4, R 5 0.99, showing a high correlation with the conven-
tional clinical urine glucose analyzer. Especially, the deviation of the measured
values in the low concentration region is small, although the deviation is medically
regarded as a problem. However, Fig. 1.4 demonstrates sufficient results for perfor-
mance of a compact and simple measuring instrument. As well, those results
showed that as a self-measuring tool at home, its portability is a plus, and it can
measure urine glucose with high accuracy with a small size of 210 mm in total
length [8].
8 Chemical, Gas, and Biosensors for the Internet of Things and Related Applications

Figure 1.4 Correlation between urine glucose meter and clinical urine glucose analyzer.

1.5 Clinical application of urine glucose meter

1.5.1 Relationship between the amount of boiled rice and urine
glucose concentration in impaired glucose tolerance
Meal load test was conducted on subjects judged to have impaired glucose tolerance
by 75 g oral glucose tolerance test. In the method, blood glucose level and urine
glucose level up to 3 hours after starting a meal were measured for 320 kcal salad
and meat, boiled rice with different amount of 100
300 g (145
435 kcal). The
blood glucose level was measured using a self-monitoring blood glucose meter
(GLUCOCARD: ARKRAY), and the urine glucose concentration was measured
with a developed urine glucose meter.
Fig. 1.5 (A) shows changes in blood glucose concentration, and (B) shows
changes in urine glucose concentration. The results confirmed an increase in urine
glucose concentration with the rise in blood glucose concentration reflecting the dif-
ference in the amount of boiled rice. In particular, differences in urine glucose con-
centrations of 400 and 600 mg/dL are difficult to determine with conventional urine
glucose test paper. The above results indicate that the quantitative measurement
using the urine glucose meter can accurately capture postprandial hyperglycemia
that changes depending on the carbohydrate intake [9].

1.5.2 Results of urine glucose monitoring on impaired glucose

tolerance case
The results of urine glucose measurement before and after a meal for 7 days are
shown in Fig. 1.6. In this impaired glucose tolerance case, self-monitoring of
Another random document with
no related content on Scribd:
That particular night—early in the evening—she had gone to Mr.
Zzyx's apartment with Henry. She happened to be carrying a new
novel which she was particularly anxious to begin reading that night.
Mr. Zzyx took a fancy to the book, probably on account of the picture
of an African jungle luridly depicted in colors on its cover. Pat refused
to give it to him, which put him in a bad temper.
She was in a dressing-gown and mules, when she discovered that,
after all her trouble in holding on to the book, she had left it behind in
Mr. Zzyx's apartment. She hurried back just as she was, and
knocked at the door. Niki answered her knock, and on her request,
returned the novel, and then closed the door. She went back to her
own apartment, and was just about to re-enter it, when she glanced
back, down the hall.
Mr. Zzyx was peering out of the doorway of his bedroom. This gave
her quite a start, and she darted into her room, quickly locking the
door after her.
Her story of what followed was an odd one.
"I wasn't so awfully frightened when I saw Mr. Zzyx peeping out at
me," she said, "or I would have gone straight into Aunt Jane's
apartment, the safest place in the castle. I had become so
accustomed to Mr. Zzyx's antics—he's just as playful as a child—I
saw no reason why I should become unduly alarmed. So I settled
down, and read my novel until about midnight. I went to sleep almost
as soon as my head touched the pillow.
"I don't know how long I'd been asleep when something aroused me.
It was a sound outside my bedroom door. I switched on the lights,
slid out of bed, caught up my dressing-gown and went to the door
and listened. I distinctly heard a scratching noise outside my door—a
sound my pet poodle makes when it wants to come into my room.
Then I saw the brass knob, inside my door, moving, and I got the
impression that some one was pressing his full weight against the
door. Well, I was just too scared to scream, so I started hammering
on the door.
"Then I listened again, by placing my ear close against the door. I'm
sure I heard a stealthy movement outside, a soft, cat's-foot
movement, as though some one was moving away, down the hall;
then everything became quiet. Finally, I became more composed
myself, and finding that I had not aroused anyone by hammering on
the door, I went back to bed. But I never closed my eyes again that
night."
When Pat recounted her adventure the next morning at breakfast,
Henry was inclined to dismiss it as trivial. "My dear, you had a
nightmare," he said. "Who on earth would want to get into your
apartment at that hour of the night? As for Mr. Zzyx, why, he wouldn't
hurt a fly."
Jane was stunned by Pat's story, and immediately added another
bolt to her bedroom door. For myself, I had heard no sound during
the night, and I'm a very light sleeper, and easy to waken. I felt, like
Henry, that perhaps Pat may have dreamed it.
At least, that was my opinion, until shortly after breakfast, while
examining the outside of her bedroom door, I found several distinct
marks, where the paint had been scratched, or clawed, off.
Discovering these marks, I felt it was not a nightmare of Pat's. So I
questioned Niki.
"You don't think it could have been Mr. Zzyx at Pat's door?" I asked
him.
Niki looked startled, then he grinned, and established a complete
alibi for our guest. "No," he said. "Mr. Zzyx never left his room last
night."
I left him, my mind confused in many ways, but entirely clear on one
point. There was something at Pat's door, that was sure.
Jane was naturally upset and uncomfortable the first night Mr. Zzyx
dined formally with us. We were both dressed for dinner, and waiting
in the entrance hall for the others to come down. She had just been
telling me of her resolve not to close the castle on November first,
and open our town house, a custom we had rigidly followed for so
many years, when she happened to glance up the grand staircase.
Clutching my arm suddenly, she whispered: "My God, Livingston!
Look!"
I turned, and gave one look, and then I burst out laughing, the sight
that met my eyes was so incredulous. Marching down the staircase,
three abreast, came Henry and McGinity, with Mr. Zzyx between
them, all three in immaculate dinner dress and enveloped in an
atmosphere of complete dignity. Henry seemed more astounded
than affronted at my mirth. Before he could voice his sentiments one
way or the other, an excited murmur came from Pat, who had just
entered the hall from the drawing room.
"Good work, Uncle Henry!" she exclaimed, rushing over to the foot of
the staircase. "And Mr. Zzyx! He looks as smart as they make them!"
Mr. Zzyx seemed pleased at the furore he was causing, and proud of
his tailed dinner clothes. He kept drawing our attention to his pearl
shirt-studs and cuff-links.
"Now, I suppose Mr. Zzyx will be expected to escort me into dinner,"
said Pat.
"Not a chance," said McGinity, moving quickly to her side; "unless
he's smarter than I think."
As Mr. Zzyx advanced towards Jane, she walked away. She stopped
when Henry said: "Don't be afraid, Jane. Mr. Zzyx has no idea of
hurting anyone. What interests him is that gold-beaded bag you're
carrying."
"Oh!" said Jane, flushing. Then she handed the bag to Mr. Zzyx, who
inspected it closely, smelt it, and then gave it back to her.
"Really!" she exclaimed. "He acts almost human!"
At this stage of the proceedings, Schweizer appeared on the scene,
to announce that dinner was served. And to my dying day, I shall
never forget the mingled look of amazement and horror that spread
over the butler's rotund face as his eyes fell on Mr. Zzyx.
And one couldn't blame him. It was enough to give any one the jitters
to see this half-human creature in smart evening dress, his heavy
animal fur framing a human face, and his hairy hands and forearms
protruding from the white cuffs of a stiff-bosomed shirt.
We had not been seated long at the table before I realized that Mr.
Zzyx, despite his repulsive appearance, possessed the mentality and
playful urge of a child. This was evident during the entrée course,
when he began to make wig-wag signals with his napkin, in an effort
to attract the butler's attention. What possible motive could he have?
Then, suddenly, the truth dawned on me. It was Schweizer who
served the dinner, and it was the food he brought that interested Mr.
Zzyx most. He had a most voracious appetite.
He sat between Henry and McGinity, in a great throne-chair which
Henry had brought from Europe. I must admit Niki had accomplished
wonders in teaching him how to handle his knife, fork and spoon. He
ate everything that was set before him, and showed a great
fondness for Henry's choice wines and champagne. Now and then,
he would pause in his eating, and look round the table, his sharp
black eyes taking us all in, one at a time; then he would chatter
something unintelligible, and resume his eating.
Henry noted this, and remarked: "Of course, we all appear very
strange to our honored guest, as it would be if we, ourselves, were
catapulted to Mars in a rocket, and suddenly found ourselves dining
with a group of Martians. In time, I hope he will be able to speak our
language."
"And then we'll know what he thinks about us," McGinity suggested.
"But he must realize by this time, how much we all think of him."
Turning to Mr. Zzyx, he patted him on the arm, and added: "You're in
pretty soft, aren't you, young fellow?"
And to out utter amazement, Mr. Zzyx turned to McGinity and spoke
—actually spoke for the first time. He distinctly mouthed a word that
sounded like "Spaghet!" with emphasis on the last syllable. He sort
of hissed the word.
"There you are!" exclaimed Henry. "I thought all along he had the
power of speech. I shall engage a tutor for him the very first thing
tomorrow morning."
"He certainly said something," McGinity observed; "sounded like
Latin to me."
And then Pat distinguished herself. "My opinion is that he tried to say
'spaghetti'," she offered. "That has some Latin connection, hasn't it?
Niki says he's terribly fond of it."
After dinner, Mr. Zzyx lounged indolently in the largest easy chair in
the library, while Schweizer served coffee. He smoked one cigarette
after another with evident enjoyment. When Henry first offered him
one of his big cigars, he surprised and amused us by biting off the
end of it, and then throwing the cigar away. The end he thrust in his
mouth and began chewing it.
A little later, as I placed my empty coffee cup on the butler's tray—
this was after Niki had taken Mr. Zzyx upstairs, to undress him and
put him to bed—Schweizer whispered: "I beg pardon, sir, but I don't
like the looks of that fellow!"
"I'm afraid, Schweizer," I rejoined, "that you'll be seeing a good deal
of this 'fellow' from now on."
The butler reflected a moment in silence on this information, and
then walked away, muttering: "I don't like his looks—I don't like his
looks!"

XVIII
The following morning, while Henry was making arrangements about
engaging a tutor for Mr. Zzyx, and McGinity busied himself in giving
proper publicity to our guest's first attempt at speech, Pat and I
strolled down to our dock. We went there on Niki's pressing invitation
to see the progress Mr. Zzyx was making in operating a runabout
Henry had recently acquired.
When we arrived at the dock, Mr. Zzyx was seated in the bow, at the
steering-wheel, looking very nautical and important in a blue worsted
suit, a white, soft-collar shirt, with a blue and white polka dot tie, and
a smart yachting cap. He beckoned at once to Pat to come down
and get into the boat.
"Oh, no! Thanks!" she called down to him.
He looked up at Pat and me imploringly. "I guess he wants us both to
come for a ride," I said. But Pat said she didn't want to go.
What happened, then and there, was an exhibition of handling a
runabout I didn't believe possible in a creature of such low mental
caliber. He seemed to take to it instinctively. Knowing there was a
great scarcity of water on Mars, I wondered how it came to him so
easily.
At Niki's word of command, he started the engine, and then steered
the boat, as unerringly as an arrow, in a swift and successful quarter-
mile run between the dock and our tiny island of rock, on which
stands the ruins of the old, stone lighthouse.
Pat and I applauded his feat on his return. In fact, Pat became so
enthusiastic over his expertness in steering the boat, that she took
her courage in hand, and ran down the steps, and jumped into the
runabout. "Now, Mr. Zzyx," I heard her say, "please give me a ride to
the island, and back."
Before I could voice my objection, Niki hopped out, and I was
horrified to see the boat race off again towards the island. While I
was protesting to Niki, I was keeping my eyes trained on the
runabout, which had now reached the island. Even from that
distance, I could see Mr. Zzyx doing a peculiar thing.
He had pulled up alongside the small dock; the engine was still
running, and the propeller kicking up a lot of foam. Apparently, he
didn't know how to tie up the boat. He was standing up, and making
funny motions to Pat, who seemed to be protesting by gestures. To
say I was not only puzzled but frightened is rather to understate the
situation.
If I had any coherent thoughts at all, they were that Mr. Zzyx wanted
Pat to go ashore with him and explore the island; he had an
abnormal sense of curiosity. There was really nothing of interest to
see there. It was all rock, devoid of trees and grass. The only
habitable building was a small shanty, which the Government had
used for storage purposes before Henry purchased the island.
"Why, in heaven's name, did you let him take Pat off like that?" I
protested to Niki. "Why didn't you stay aboard? He's dangerous."
Niki shrugged rather insolently at my fears. Since receiving a liberal
payment from Henry on the reward he had earned by first
discovering the rocket, he had become rather impudent. "Mr. Zzyx is
not dangerous, Meester Livingston," he countered. "He only likes fun
—like a leetle kitten!"
"When I say he's dangerous I mean that he is," I replied, with
vehemence. Then, profoundly shaken over Pat's security, I cried:
"Oh, what shall I do?" and turned round to run back to the castle.
The next moment, I had run full tilt into McGinity.
McGinity's reaction to the situation was typical. "That bird is too
dangerous to trifle with," he said; and within a few minutes he was in
one of our row-boats, heading for the island.
To my amazement, the next thing I saw was Mr. Zzyx bringing the
speed-boat back to the dock. I had pulled myself together somewhat
when he returned. I was surprised to see Pat, lolling comfortably
among some cushions, a cigarette in her hand and a cool smile on
her face. By that time, McGinity had put back to the dock. He arrived
in time to help Pat out of the runabout.
"What's all the trouble?" she asked him, a little sharply.
"Oh, I don't know," he replied, vaguely. "We just thought you might
be stuck over there, and couldn't get back."
"No fear," she returned, and laughed a little. "Mr. Zzyx has too much
common sense for that. The way he handles the boat is simply
marvelous. It was real excitement. I wouldn't have missed the ride—
not for anything!"
"All the same," McGinity remarked, "it was lucky for you, perhaps,
that I got here in time—that is, in case anything did happen."
Pat waved that off with a light gesture, and turned to me. "How does
Uncle Livingston feel about it?" she inquired.
"I was pretty well excited myself, Pat," I replied; "uneasy-like."
"I simply wanted to show Mr. Zzyx that I'm not afraid to be alone with
him, and that I'm a good sport besides," Pat explained.
"It's bad business," said McGinity, "any way you look at it. Matter of
fact, I think Mr. Zzyx is bad business."
"I don't see it," Pat retorted.
It was not until an hour later, when I found myself alone with Pat, that
I learned the truth about the situation, although I was convinced from
the first that she had deliberately overplayed her attitude of
indifference to danger.
"I was nearly scared to death," she confessed. "I did not miss Niki
until we were well on our way to the island. Mr. Zzyx showed by his
actions that he wanted me to tie up the boat, and go ashore with
him, but I wouldn't budge. If he had remained there, at the dock, a
minute longer, I would have yelled, screamed—jumped overboard.
But—" she concluded, as she tightened her grasp of my arm, "never
mind what I felt—Bob mustn't suspect."
In other words, she was still concealing her fear of Mr. Zzyx lest
something might happen to spoil McGinity's news stories, and
remove him from our midst. For some weeks now, he had been
assigned by his paper to "cover" Mr. Zzyx's every movement, and to
report all the news developments in connection with Henry's theories
about Mars. This necessitated his remaining at the castle.
The Daily Recorder, however, long since, had announced in its
editorial columns that while it printed all the news concerning the
recent Martian revelations, the publishers assumed no responsibility
for their veracity, and their readers were left to render a verdict in
accordance with the facts. This was a little raw on Henry, I thought.
Anyhow, the incident of the runabout was forgotten in the excitement
of the following day, when the Swedish-born Mayor of New York, His
Honor, Oscar Swenson, gave an official reception for Mr. Zzyx at the
City Hall. And what happened there became local history.
There was a large crowd outside our lodge-gate as we drove off, in
an open car; and all along the way, in the suburbs, and through the
city, to the downtown district, police reserves had to be called out to
control the vast throngs which lined our route.
It was a triumphal procession through the city. Mr. Zzyx waved to the
people in response to their loud huzzas. From the clouds of ticker-
tape and confetti that descended upon us, he collected a great
quantity. He and Henry, sitting in the rear seat, were knee-deep in it
by the time we reached the City Hall. On our way, while the
procession was held up by cross-town traffic, I bought a raspberry
lollypop from a street vendor for Mr. Zzyx. Henry frowned on this as
very undignified, but Mr. Zzyx sucked it with great enjoyment. His
actions reminded me of a small boy at his first circus.
The day was perfect—the air cool and crisp. We found City Hall
plaza one vast sea of faces. As we passed through a barrage of
cameramen, a Swedish chorus burst into song; and we had no
sooner taken our places on the steps, beside the Mayor, scores of
other city officials, and many notable invited guests, when several
hundred Swedish gymnasts entertained us with feats of physical
prowess.
I wished that Jane and Pat had come with us, but they both had
elected to remain at home, and enjoy the happenings at the City Hall
through the medium of the radio and television.
Mayor Swenson is a tall, gaunt, rosy-cheeked Swede, but his head
only reached to the shoulders of Mr. Zzyx when they stepped in front
of the microphone and television transmission instruments. Niki had
accompanied us as the Martian's bodyguard, and never left his side.
He carried an automatic pistol, ready for any emergency, as I
learned afterwards.
It was not my first experience of an official reception at the City Hall,
but many years had intervened since I attended the last one.
Although I was very familiar with the great changes that had taken
place, politically, in the city administration, this first close personal
contact with the Mayor, the Board of Aldermen, and the various
Commissioners, was in the nature of a shock. There was not one
single Irishman in the Aldermanic board, nor even a Jew. The board
was composed mostly of Chinese, Turks, Filipinos, and Bulgarians,
and one Eskimo, who had entered politics after graduating with
honors from Princeton University.
Amid this gathering of mixed nationalities, Mr. Zzyx was an
outstanding figure. As time passed, he grew restless, and kept
running a long, hairy finger around his immaculate collar as though it
choked him. He was attired in a formal cutaway coat and striped
trousers, topped with a silk hat, which he wore at an angle that gave
him a rather rakish appearance. He looked to me exactly like a huge,
over-stuffed piece of furniture, with the hair sticking out.
A breathless hush fell upon the thousands as the Mayor raised his
hand to command silence. His voice sounded a bit squeaky through
the loud speakers, not thunderous, as I had anticipated. I took notes
of his speech, which follows:
"I t'ank it's about time I introduced the city's distinguished visitor from
Mars." (A pause until the cheering had subsided.) "I bane t'anking as
I stood here that Mr. Zzyx is probably the most unique visitor the
great city of New York has ever welcomed, officially, yah?" (More
applause.) "Some of you no doubt bane t'anking that he is a great
fakir. My wife and me, we bane having an argument about this. My
wife, she t'anks he's just a big monkey that's got loose from some
zoo." (Laughter.) "When I ask our good friend, Mr. Henry Royce,"
(Mayor's voice lost in a tumult of cheers)—"when I ask him what he
bane t'anking about Mr. Zzyx, and his coming in a rocket, on a beach
out on Long Island, he only shakes his head, and says he knows
next to noddings; and I t'ank he's just as much fooled as the rest of
us. But whether our distinguished guest comes from Mars or the
moon, I bane t'anking we must hold fast to our traditions, and bestow
on him the key to our great city of fifteen million people—yah?
Therefore, it is with the greatest pleasure that I confer such an honor
upon Mr. Zzyx, the jungle man from Mars."
So saying, the Mayor handed an important-looking scroll to Mr. Zzyx,
who took it, and immediately unfolded it and began to look for
pictures. Not finding any, he passed it over to Henry, at whose signal,
Niki stepped forward to superintend Mr. Zzyx's introduction at the
microphone.
Our Martian visitor made a better showing at the microphone than I
had expected. I did not know then that for several days previous to
the reception, Niki had coached him in the use of the instrument.
First, he peered curiously into it, then he stuck his finger in, as
though he had seen some imprisoned insect inside.
Suddenly, he began to chatter, and then, just as suddenly, he
stopped. Hearing his own voice amplified through the loud speakers
seemed to have startled him. After Niki had patted him reassuringly
on the arm, he burst into chatter again, concluding with the only word
he could pronounce—"Spaghet!" He seemed to spit the word into the
microphone, which sent the crowd into convulsions of laughter and
cheers.
That practically ended the official city reception. After stepping into
our car, Mr. Zzyx further amused the crowd by smoking a cigarette,
and tipping his hat to the ladies, another trick Niki had taught him.
On the return trip to Long Island, he was greeted with even greater
acclaim than had been shown him earlier in the day.
We found Pat in a state of excitement. Mrs. Cornelius Van Dyk, she
said, had telephoned during our absence, to announce that she was
giving a dinner for Mr. Zzyx on the following Monday. After dinner,
she planned to take Mr. Zzyx to the opening performance of the
winter season at the Metropolitan-Civic Opera House.
Naturally, Pat was excited about this; we all were. Mrs. Van Dyk is
the last word in fashionable exclusiveness in New York society; even
European royalty is more accessible.

XIX
Mr. Zzyx behaved beautifully at the very brilliant dinner given in his
honor by Mrs. Cornelius Van Dyk at her town residence, the last red
brick mansion of a remote period, except our own, still left standing
in Washington Square. A dinner made more memorable than it
otherwise would have been by the distinguished array of guests.
Among them, Henry's beloved and revered old friend, the venerable
Episcopalian prelate, Bishop William Buckingham, who had grown a
bit queer in his dotage.
"A very novel idea on the part of our hostess," the Bishop remarked
to Henry after dinner, in the smoking-room, while I sat by, listening
and silent. "This sort of thing was done, years ago, at Newport, a
monkey-dinner, as I recall reading about it, and the clergy and the
newspapers made an awful row. Certainly times have changed when
we can sit down to dinner with a man-ape without the flicker of an
eye-lash. After this, I shan't be at all surprised to have one of my old
parishioners invite me to dine with a white rabbit. Mrs. Van Dyk sets
the fashions in New York, you know."
"After all," remarked Henry, "brutes and humans really belong to one
great family by common descent."
"Hold your miserable tongue, sir!" the Bishop responded, perkily.
"Not until I've extended an invitation to your reverence, to attend the
banquet the Exploration Club is giving for Mr. Zzyx on November
thirtieth," Henry rejoined.
"The Exploration Club! How extraordinary!" the Bishop exclaimed.
"The most exclusive club of its kind in the city. What's up?"
"Oh, just another revelation concerning Mars," Henry replied,
nonchalantly. "You will come, won't you?"
"Do my best to oblige," the Bishop replied.
At that, I felt my ears pricking. I already knew that Henry, Olinski and
McGinity, had something new about Mars up their sleeves, which
was to be disclosed at the banquet at the Exploration Club. What it
was, I had no idea. And I found out nothing that night. The
conversation between the Bishop and Henry was cut short by the
return of Mr. Zzyx and Niki, who had taken our Martian visitor to the
lavatory immediately after dinner, to tidy him up a bit before we left
for the opera.
A few moments later, we joined Mrs. Van Dyk, and her house guest,
Lady Gwynne of London, in the drawing room. The other dinner
guests had gone. Both were ready for the opera, Mrs. Van Dyk in a
stunning ermine wrap, and Lady Gwynne in sables.
We had no sooner entered the room, when, to my horror, Mr. Zzyx
went straight up to our hostess, and began to chatter, and stroke her
ermine coat. Then he walked over to Lady Gwynne, and repeated
the action on the sable wrap. I could see that they were both terribly
frightened.
Henry took the matter in hand at once, and drew Mr. Zzyx aside,
tenderly, as a father would treat a child of doubtful sanity. After
quieting him with a cigarette, he left him in Niki's care, and
approached Mrs. Van Dyk.
"A bad break, I'm afraid," he said to her, "and I apologize for this
breach of propriety. After all, Mr. Zzyx is part animal, and I'm afraid
the high instinctive animalism in him was beguiled by the sight and
smell of ermine and sable."
"A gesture of Martian jungle courtship," Lady Gwynne suggested.
Henry shook his head. "No; I don't think so," he said. "Mr. Olinski, my
associate, and I, have definitely proved that he is not influenced in
any way by what we mortals call sex appeal. Otherwise, he would be
very objectionable to have about. Pretty clothes, sparkling gems and
furs attract him just as toys intrigue small children. While instinctively
curious, and perhaps a little bold, he means no harm."
"Let's hope he'll keep up this high standard of behavior," the Bishop
remarked. "Undoubtedly a tremendous brute force lies sleeping
under his apparent docility. A pretty go, if this brute force is ever
aroused in him."
"I hope to God that'll never happen," said Henry, gravely.
And then Mrs. Van Dyk spoke. "We can't expect him to measure up
to Park Avenue social standards," she said. "A little clowning now
and then is relished by the best of men. Indeed, I've known men in
my own set to go much further than the mere stroking of a lady's fur
coat."
"Exquisite!" laughed the Bishop.
"How droll!" Lady Gwynne commented.
"As a creature from another planet," Mrs. Van Dyk continued, "I feel
very honored in having Mr. Zzyx as a guest in my house."
Henry sighed gustily, and said: "Very friendly of you, Mrs. Van Dyk."
And then the Bishop said: "Well, let's push on to the opera."
The Metropolitan-Civic Opera House was packed that night with one
of the largest crowds in its history. I was convinced upon our arrival
that the throng was there, not to hear Verdi's opera "Otello," but to
see Mr. Zzyx. The evening newspapers had heralded our coming,
and we encountered a large crowd outside the opera house, and
were met by a barrage of cameramen's flashlights as we entered.
Once inside, the crush about us was so great, we had considerable
difficulty in reaching Mrs. Van Dyk's box, in the parterre. Although
grand opera now was democratized, the "diamond horseshoe" still
remained. Opera, I'm afraid, will always remain the pet hobby of the
fashionably rich, just as racing will ever be regarded as the sport of
kings.
Two uniformed city policemen stood on guard, in the corridor, outside
the box. Mrs. Van Dyk, regal in black velvet and sparkling with
jewels, occupied the corner nearest the stage. Mr. Zzyx sat in the
other corner, with Henry sitting between. Behind them, Bishop
Buckingham was sandwiched between Lady Gwynne and Jane,
while I hovered, standing, in the rear, too nervous to sit down. Niki
was at my elbow.
We had missed the first act. Five minutes after we had settled
ourselves in the box, the curtain rose on the second act. Fashionable
women, like Mrs. Van Dyk, seem to make it a point to be late at the
opera. I doubt if our hostess had ever heard the first act of any opera
in the entire Metropolitan-Civic repertoire, during her long ownership
and occupancy of the box.
During that five minutes, every eye in the house appeared to be
turned on Mr. Zzyx, who, fortunately, was now in a state of lassitude,
which always overtook him after a heavy dinner. Apparently
undisturbed by the sensation he was causing, he devoted himself,
first, to a curious scrutiny of the packed masses in the balconies,
then he looked down at the arena below, and, finally, rested his gaze
on the two rows of boxes, filled with superbly gowned and bejeweled
women.
I was curious to see what effect grand opera music would have upon
him. What little music he had heard at the castle had come from our
radio, and in this he had displayed only a mild interest. His attitude
toward such music as he had heard rather dispelled the theory that
had been advanced, that if direct radio communication was ever
established between the earth and Mars, the interchange of ideas
would necessarily have to be through the medium of music, on
account of the lack of a common language.
Here, at last, was a chance to try music at its best on an inhabitant of
Mars. I wondered what the reaction would be. Mr. Zzyx watched the
musicians curiously as they trickled into the pit, and the noise of the
tuning up seemed to interest him immensely. Finally, when the house
went dark, he appeared quite excited. Then the baton of the
conductor rose, and the first crash of the orchestra came like a
thunder-clap.
Mr. Zzyx leapt to his feet, and started to climb over the edge of the
box. For a frenzied moment, I thought he was going to dive head first
into the midst of the spectators below. But Henry quickly grabbed
him by his swallow-tails, and pulled him back into his chair. The
incident did not attract the general attention it might have done if the
auditorium had not been darkened.
While Henry patted Mr. Zzyx on the shoulder to quiet him, Mrs. Van
Dyk leaned over, and said: "I don't wonder at him trying to jump out
of the box. To many, grand opera is a perfect hullabaloo, and
devastating. That's why so many people go out between acts for a
cocktail."
Presently Mr. Zzyx fell to listening, with his mouth open. At first, I
thought he was wholly lost in the delight of the orchestral movement
—drums and horns were silent now—and the beautiful singing on
the stage. Then, like a flash, it occurred to me that it was the dark-
skinned Otello who was claiming his attention, not the music or
singing.
I watched him, studied him attentively, as the opera swept on to its
violent climax—the smothering to death of Desdemona—by the
enraged Otello. After the final curtain, while the audience was
recalling and applauding the singers, I noticed he looked a little wild
about the eyes; a sort of inward brooding.
Was it possible that he had grasped the significance of the story, as
it had been unfolded before him on the stage? Could the climax of
the opera put ideas into his head beyond his purely natural instincts?
The force of ideas even stronger than his own inherent brute force,
which might quicken him to the fury of some deed of incredible
violence?
But I had no time for surmises. Yet, as we passed out of the opera
house, in an atmosphere of acclaim and some disorder, almost
mechanically, I jotted down the details in my memory of what I had
observed in him. From the look in his eyes, I felt some terrific instinct
had been aroused. It gave me a strange and eerie feeling, but I
made no mention of it to Henry.
Within ten days I was glad to have paid attention to such details.
Little did I suspect then that a black, threatening cloud was gathering
over our heads, or that more mystery, intrigue—even death—was
closing in about us.

XX
Life at the castle followed its usual routine during the interval
between Mrs. Cornelius Van Dyk's dinner and opera party, and the
banquet at the Exploration Club, in Mr. Zzyx's honor, with one
exception, which I shall mention further on.
I was delighted to see that McGinity shared the place of honor
allotted to our family group at the banquet, and justly so, because he
had put Henry's discoveries and theories over in the biggest possible
way. Later in the evening, I found he had other honors accruing to
him.
It was not until I was seated with the family party at the head of the
long U-shaped table, that I noticed the motion picture screen at the
far end of the dining room. Then the full significance of those secret
visits to a large film studio in Long Island City, on the part of Henry,
Olinski and McGinity, began to dawn upon me.
Gradually worming the secret out of McGinity, who sat on my left
(Jane was on my right) I was in possession of the complete facts of
the Martian revelations, shortly to be disclosed to this most highly
honored body of explorers and scientists, by the time the soup
course was over.
After Olinski had deciphered the Martian written message contained
in the mysterious scroll, found in the rocket, McGinity had put the
information contained therein into scenario form. A screen
production, backed by Henry's money, had been staged by one of
the largest and most progressive film corporations, at its Long Island
City studio, with Henry and Olinski acting in an advisory capacity.
I am telling this circumstantially, because the part McGinity played in
writing the scenario made the first real contribution to the solving of
the strange mysteries that enveloped us, and because it explains
how I myself in a small way became involved in the untangling of the
web.
As we sat placidly at the banquet table, my last thought was that
within twenty-four hours we would be plunged into a series of events,
which savored of the sort of thing associated with sensational fiction,
or exciting melodrama on the screen.
At odd moments, I cast my eye across the table at Mr. Zzyx. His
prolonged sojourn under our roof had become a "beastly vulgar
business," quoting Jane's own words. Daily, we were growing more
resentful of his impenetrable stupidity, and utterly bored with his
gross and ugly presence. Often I felt myself in the mood to wring his
neck.
It was also perfectly clear to me that Henry was beginning to tire of
shouldering the responsibility of this big, lumbering creature, but so
far he had kept it to himself. I felt angrier with him that I had ever
been in my life, yet I was angry rather for him than with him. It was
so utterly unlike him to allow the family's unpleasant associations
with Mr. Zzyx to continue, when a word from him would have ended
it.
The exception to our usual routine at the castle, during the week,
related to our guest from Mars. He was beginning to act very
queerly. I was of the opinion that a sort of madness was creeping on
him, brought on by the unnatural state in which he was living, the
strange food he ate so ravenously, and the constant excitement to
which he was subjected. One of the spookiest things he did was to
move about the castle during the night. Niki might be on guard, and
Mr. Zzyx's own bedroom door locked and bolted, but with uncanny
skill both were circumvented.
His first real outburst had come on the Friday night, preceding the
banquet. He began throwing things at Niki, and did considerable
damage to the furniture, pictures and walls in the State Apartment.
When I questioned Niki, he had dismissed the affair lightly, with the
excuse that Mr. Zzyx had been suffering from insomnia, and was not
himself.
Certainly he was not himself at the Exploration Club banquet. During
the dessert course, I saw that he had not touched his charlotte
russe, and was making holes in the table-cloth with his fork. His pet
hobby, while dining, was to roll his bread into little balls, toss them up
in the air, and then catch them in his mouth as they fell, something I
considered inexpressibly vulgar and disgusting.
I was astonished that Henry, or Bishop Buckingham, who was a
member of our party, did not rebuke him for making holes in the
cloth; but both seemed preoccupied. In a state of anxiety, I glanced
around at Pat, who was sitting on McGinity's left. It was not strange
to find that they both were practically oblivious to their surroundings.
The speech-making was now going on, having begun shortly before
coffee was served. The speakers were long-winded and tiresome. I
am neither a student, nor a philosopher, but I would like some
exponent of the doctrine of psychology to explain why men talk so
much and at such great length at banquets. I've often wished that
some bright person would organize a society for the suppression of
after-dinner speakers.
For fully half an hour, now, a little, rabbit sort of man, with big ears
and completely bald, and wearing tortoise-shell spectacles, had
been telling of his pursuit of prey, biped and quadruped, in distant
places, with minute detail of how he had killed one of every species
of beast and bird and fish in the world. The guests were showing
signs of impatience. Mr. Zzyx began making horrible grimaces, when
Henry tapped him warningly on the arm. Then he started to amuse
himself making those little bread balls. I became uneasy myself for
fear he might throw one of them at the speaker, something I wanted
to do myself but did not dare.
Then, suddenly, to my stunned astonishment, Mr. Zzyx picked up the
untasted charlotte russe, which is custard in a form made of sponge
cake, and hurled it at the speaker, who was directly opposite him.
His aim was true, and the little rabbit man got the charlotte russe full
in the face.
The guests roared with delight as the mighty hunter dug his features
out of the spattering custard, while Henry shook Mr. Zzyx sternly by
the arm, and whispered: "You ought to know better!"
Bubbling with mirth, I leaned over to McGinity, and said: "Too bad he
got it in the face." To this, the reporter replied: "The main thing is that
he got it."
After the bespattered speaker had gone to the lavatory to wash his
face, the toastmaster rose, and said: "Now that Mr. Zzyx, our
honored guest from Mars, has enlivened our dinner, we shall
proceed to the surprise event of the evening.
"No one is asked to accept these new disclosures about Mars which
our friend, Mr. Olinski, decoded from the mysterious writings of the
scroll, discovered in the rocket, as infallible," he continued. "Even our
fellow-scientist, Mr. Royce, who is accountable for this, and other
recent events of a scientific nature, which literally have rocked the
world, declares an uncertainty still exists in his own mind, and that
he is simply making public the information that has fallen into his
hands, from strange and unknown sources. In other words, he
wishes me to make clear to you all that he's not trying to put
something over on us. So, now, let's see what we shall see!"
The film, in four reels, was in the nature of a travelogue, beautifully
colored, and interspersed with sound and music. Henry was the
pictorial lecturer. McGinity's clever hand was seen in the numerous
whimsies and dramatic highlights. Many scenes were genuinely
stirring.
Mr. Zzyx, closely guarded by Niki in the darkened dining room,
watched the picture unfold with fascinated interest. At times, he
would gesticulate, strangely, like one familiar with the subject matter,
and utter primitive sounds, as though he wanted to speak, and tell us
more startling things about his home planet.
This newly acquired and first-hand information of present day life on
Mars, presented in picture form, supplemented by the free play of
imagination on the part of the director, proved infinitely more
valuable as educational entertainment than the cold facts would
have been if delivered from the lecture-platform.
The picture divulged, first of all, that life on Mars had originated and
evolved the same as on the earth, with the white division of the
human species exercising supreme authority over the affairs of the
planet.
Secondly, it showed that the strange, geometric markings on the
planet, as studied by astronomers on earth, are not a canal system,
or even man-made. The lines, or bands, which some of our
astronomers believed to be canals, constituting a system of
irrigation, are really deep wide canyons, ten to twelve miles in width
at the rim, and descending 2,000-3,000 feet below the sterile