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Enhanced electrochemical detection of tartrazine in beverages and liquid soap via nickel phosphide-adorned functionalized carbon nanofibers

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Journal of the Taiwan Institute of Chemical Engineers 2024年 157卷

作者： Gokulkumar, Kumar Huang, Song-Jeng Kogularasu, Sakthivel Aljuwayid, Ahmed M. Maheshwaran, Selvarasu Govindasamy, Mani Department of Mechanical Engineering National Taiwan University of Science and Technology Taipei City106335 Taiwan Super micro mass research and technology center Cheng Shiu University Kaohsiung833301 Taiwan Center for Environmental Toxin and Emerging-Contaminant Research Cheng Shiu University Kaohsiung833301 Taiwan Department of Chemistry College of Science King Saud University P. O. Box 2455 Riyadh11451 Saudi Arabia Department of Aeronautics and Astronautics National Cheng Kung University Tainan701 Taiwan International Ph.D. Program In Innovative Technology of Biomedical Engineering and Medical Devices Ming Chi University of Technology New Taipei City243303 Taiwan Tamil Nadu Chennai602105 India

Background: Tartrazine (TAT) is a widely used food colorant dye, offering an orange-yellow hue in various products such as foods, cosmetics, and pharmaceuticals. Despite its popularity, TAT is banned in several countries due to its potential health risks, including respiratory issues, migraines, and skin allergies. Moreover, TAT is linked with adverse effects on human physiology, including mutagenicity, biliary cirrhosis, changes in liver and kidney functions, and neurodegeneration at varying doses. Consequently, the detection of this food additive is crucial for safeguarding human health and protecting aquatic organisms. Methods: In this study, a novel sensor was developed for the detection of TAT, utilizing a composite material made from nickel phosphide decorated with functionalized carbon nanofiber (Ni2P@f-CNF). The Ni2P@f-CNF was synthesized and characterized through several techniques, including X-ray diffraction, X-ray photoelectron spectroscopy, field emission-scanning electron microscopy, and elemental mapping. The composite was then integrated with a glassy carbon electrode to create a sensor. The sensor's performance was evaluated in terms of detection limit, linear range, selectivity, sensitivity, reproducibility, repeatability, and long-term stability. Main Findings: The developed Ni2P@f-CNF composite sensor displayed a lower detection limit of 0.011 µM and a broad linear range from 0.01 to 1875 µM. Its selectivity and sensitivity were exceptional, primarily due to the catalytic activity of the Ni2P@f-CNF composite towards TAT. The sensor also demonstrated excellent reproducibility, repeatability, and notable long-term stability. Importantly, when applied in real-world scenarios, the sensor successfully detected TAT in soft drink and liquid soap samples, yielding high recovery rates between 97.45 % to 99.37 % and 98.26 % to 99.01 %, respectively, underscoring its potential applicability in practical settings. © 2024 Taiwan Institute of Chemical Engineer

关键词： Nickel compounds

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biomedical data and AI

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science China Life sciences 2025年第5期68卷 1536-1540页

作者： Hao Xu Shibo Zhou Zefeng Zhu Vincenzo Vitelli Liangyi Chen Ziwei Dai Ning Yang Luhua Lai Shengyong Yang Sergey Ovchinnikov Zhuoran Qiao Sirui Liu Chen Song Jianfeng Pei Han Wen Jianfeng Feng Yaoyao Zhang Zhengwei Xie Yang-Yu Liu Zhiyuan Li Fulai Jin Hao Li Mohammad Lotfollahi Xuegong Zhang Ge Yang Shihua Zhang Ge Gao Pulin Li Qi Liu Jing-Dong Jackie Han Peking-Tsinghua Center for Life Sciences (CLS) Academy for Advanced Interdisciplinary StudiesPeking University Center for Quantitative Biology (CQB) Academy for Advanced Interdisciplinary StudiesPeking University Peking University-Tsinghua University-National Institute of Biological Sciences Joint Graduate Program Academy for Advanced Interdisciplinary StudiesPeking University Department of Physics University of Chicago School of Life Sciences Southern University of Science and Technology Peking University Chengdu Academy for Advanced Interdisciplinary Biotechnologies College of Chemistry and Molecular Engineering Peking University Department of Biotherapy Cancer Center and State Key Laboratory of BiotherapyWest China HospitalSichuan University Department of Biology Massachusetts Institute of Technology Lambic Therapeutics Inc. Changping Laboratory Al for Science Institute Key Laboratory of Computational Neuroscience and Brain-Inspired Intelligence Fudan University Department of Obstetrics and Gynecology West China Second University HospitalSichuan University Peking University International Cancer Institute and Peking University-Yunnan Baiyao International Medical Institute and State Key Laboratory of Natural and Biomimetic Drugs Department of Molecular and Cellular PharmacologySchool of Pharmaceutical SciencesPeking University Health Science CenterPeking University Channing Division of Network Medicine Department of MedicineBrigham and Women's Hospital and Harvard Medical School Center for Artificial Intelligence and Modeling the Carl R.Woese Institute for Genomic BiologyUniversity of Illinois Urbana-Champaign Department of Genetics and Genome Sciences School of Medicine and Department of Computer and Data Sciences and Department of Population and Quantitative Health SciencesCase Western Reserve University Department of Biochemistry and Biophysics University of California Sanger Institute Department of Automation Tsinghua University State Key Laboratory of Multimodal Artificial Intelligence Systems I

The development of artificial intelligence(AI) and the mining of biomedical data complement each other. From the direct use of computer vision results to analyze medical images for disease screening, to now integrating biological knowledge into models and even accelerating the development of new AI based on biological discoveries, the boundaries of both are constantly expanding, and their connections are becoming closer.

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Muscle coactivation analysis for neuromuscular control assessment of lower limb stretch-shortening cycle

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Gait & Posture 2023年 106卷 S175-S176页

作者： Rodrigues, Carlos Correia, Miguel Abrantes, João Benedetti, Marco Nadal, Jurandir Faculty of Engineering- University of Porto Biomedical Engineering Program Porto Portugal INESC TEC - Institute for Systems and Computer Engineering- Technology and Science C-BER - Centre for Biomedical Engineering Research Porto Portugal Faculty of Engineering- University of Porto Department of Electrical and Computer Engineering Porto Portugal CICANT - Centre for Research in Applied Communication- Culture and New Technologies MovLab - Laboratory of Technologies for Interactions and Interfaces Lisbon Portugal Federal University of Pernambuco Department of Electronic and Systems Recife Brazil Federal University of Rio de Janeiro Biomedical Engineering Program Rio de Janeiro Brazil

IntroductionMuscle stretch-shortening cycle (SSC) is a central mechanism with lower limb muscle contraction immediately preceded by muscle stretch for efficient submaximal activities such as gait and powerful maximal activities such as running and jumping [1,2]. Research questionAlthough muscle SSC can be observed at gait and running its higher expression and accessibility is performed on standard maximum vertical jump (MVJ) with an open issue on neuromuscular control assessment of lower limb muscle SSC [3]. MethodsFor this purpose, we present and applied noninvasive subject specific analysis of lower limb muscle coactivation for selected muscles with higher contribution during MVJ impulse based on corresponding conditioned surface electromyographic signals (sEMG). Selected muscles correspond to lower limb muscles vastus medialis (VM), rectus femoris (RF), vastus lateralis (VL), lateral gastrocnemius (LG) and medial gastrocnemius (MG). Twenty-seven trials were assessed corresponding for each subject to the highest MVJ based on larger flight time with long SSC at countermovement jump (CMJ), short SSC at drop jump (DJ) and squat jump (SJ) with no SSC. Trial sample is composed by a group of six young adult volunteers’ students on sports and physical education degree with the ages (21.5 ± 1.4) years, (76.7 ± 9.3) kg mass and (1.79 ± 0.06) m height. Surface skin was prepared and Aqua-Wet gel Skintact F55 electrodes were applied at bipolar configuration as indicated by SENIAN. VM, RF, VL, LG and MG sEMG linear envelopes were paired plotted with the area under the curves computed as well as the coactivation pairs defined as the common area under the curves normalized to the sum of the corresponding pair individual areas under the curves. Coactivations were compared at CMJ, DJ and SJ as well as among CMJ, DJ and SJ, Fig. 1. ResultsStrongest coactivations with decreasing intensity were detected between VM-RF and LG-MG muscles at CMJ and SJ, whereas at DJ the strongest coacti

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Linearity assessment between lower limb joint angles and angular accelerations at standard maximum vertical jumps with long-short and no countermovement

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Gait & Posture 2023年 106卷 S174-S175页

IntroductionStandard maximum vertical jump (MVJ) has been applied to assess lower limb long countermovement (CM) on countermovement jump (CMJ) and short CM on drop jump (DJ) for comparison to squat jump (SJ) without CM [1,2]. Research questionNevertheless, the ability for lower limb joint accelerations depending on lower limb joint angles at long, short and no CM remains an open issue [3,4], in particular linearity assessment of these relations at long, short and no CM in association with MVJ whole-body impulsion. MethodsFor this reason, we applied piecewise linear detection of segmented subphases on lower limb joint angles ( θ)-angular acceleration ( α) diagrams for the hip (H), knee (K) and ankle (A) during impulse phases on CMJ, DJ and SJ. Selected sample corresponds to best MVJ trials based on higher flight-time of 3 CMJ, DJ and SJ repetitions for six healthy untrained subjects with ages (21.5±1.4) years, (76.7±9.3)kg mass and (1.79±0.06)m height. Adhesive reflective markers were applied at lower limb and torso, with sagittal hip, knee, and ankle θ, α obtained by inverse kinematics. H,K,A ( θ, α) diagrams were plotted and impulsion subphases segmented at selected instants of ∂ a/∂ q phase reversal with linear fit by the least-squares method, Fig. 1. Fig. 1. Hip, knee, ankle joint ( θ, α) diagrams at CMJ, DJ, SJ with segmented subphases time span(Δt), |∂α/∂θ| slope magnitudes and R² linear fit regression. ResultsSJ presented larger number of ( θ, α) subphases than DJ, both larger than CMJ with higher |∂ α/∂ θ| variation at DJ and SJ than CMJ, pointing to higher complexity of neuromuscular control and contributing to explain reduced SJ, DJ performance without CM (0.33±0.05)m and with short CM (0.27±0.03)m in relation to CMJ with long CM (0.36±0.04)m for untrained selected subjects, Fig. 1. Thus, despite the larger total time span Δt at CMJ (0.76±0.10)s in relation to SJ (0.36±0.08)s and DJ (0.23±0.05)s, CMJ presented lower number of subphases with longer time span

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Electrochemical biosensor based on an atomic layered composite of graphene oxide/graphene as an electrode material towards selective and sensitive detection of miRNA-21

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Microchemical Journal 2024年 199卷

作者： Huang, Min-Shin Govindasamy, Mani Chinnapaiyan, Sathishkumar Lin, Yi-Ting Lu, Shao-Yang Samukawa, Seiji Huang, Chi-Hsien Department of Materials Engineering Ming Chi University of Technology New Taipei City 243303 Taiwan International Ph.D. Program in Innovative Technology of Biomedical Engineering and Medical Devices Ming Chi University of Technology New Taipei City 24303 Taiwan Centre for Applied Research Saveetha School of Engineering Saveetha Institute of Medical and Technical Sciences (SIMATS) Chennai-602105 Tamil Nadu India Institute of Fluid Science Tohoku University Sendai 980-8577 Japan College of Electrical and Computer Engineering National Yang Ming Chiao Tung University Hsinchu 300093 Taiwan Division of Rheumatology Allergy and Immunology Chang Gung Memorial Hospital Taoyuan City 33305 Taiwan College of Engineering Chang Gung University Taoyuan City 33302 Taiwan

Development of an efficient technique towards the detection of miRNAs is highly significant for the timely diagnosis and prognosis of cancers. In this study, we utilized the chemical vapor deposition (CVD) technique to grow bilayer graphene (BLG) on copper foil and subsequently transferred the BLG to fluorine doped tin oxide (FTO) substrate via a facile wet transfer technique. The upper layer of the BLG was modified to graphene oxide (GO) with a low damage plasma treatment (LDPT) process to serve as a biomolecule receiving layer and the lower graphene layer (G) serves as an electron transport layer. The developed GO/G/FTO platform offers superior sensing ability towards the miRNA-21. The results demonstrated that the proposed GO/G/FTO substrate detect the miRNA-21 with a wide linear range (10 fM ∼ 1 nM), low limit of detection (3.18 fM), high sensitivity (0.6 mApM⁻¹cm⁻²), selectivity and stability via the differential pulse voltammetry method. Additionally, the sensors can efficiently discriminate the target miRNA-21 from its complementary base mismatched counterparts. The proposed approach serves as an excellent tool for the early diagnosis of diseases. © 2024 Elsevier B.V.

关键词： Atomic layer oxidation Biosensor Cancer biomarkers Electrochemical Graphene oxide Plasma treatment

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M2FN: Multi-step modality fusion for advertisement image assessment

arXiv

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arXiv 2021年

作者： Park, Kyung-Wha Ha, Jung-Woo Lee, JungHoon Kwon, Sunyoung Kim, Kyung-Min Zhang, Byoung-Tak Interdisciplinary Program in Neuroscience Seoul National University Korea Republic of NAVER AI LAB NAVER CLOVA Statistics and Actuarial Science Soongsil University Korea Republic of School of Biomedical Convergence Engineering Pusan National University Korea Republic of Department of Computer Science and Engineering Seoul National University Korea Republic of Surromind Robotics

Assessing advertisements, specifically on the basis of user preferences and ad quality, is crucial to the marketing industry. Although recent studies have attempted to use deep neural networks for this purpose, these studies have not utilized image-related auxiliary attributes, which include embedded text frequently found in ad images. We, therefore, investigated the influence of these attributes on ad image preferences. First, we analyzed large-scale real-world ad log data and, based on our findings, proposed a novel multi-step modality fusion network (M2FN) that determines advertising images likely to appeal to user preferences. Our method utilizes auxiliary attributes through multiple steps in the network, which include conditional batch normalization-based low-level fusion and attention-based high-level fusion. We verified M2FN on the AVA dataset, which is widely used for aesthetic image assessment, and then demonstrated that M2FN can achieve state-of-the-art performance in preference prediction using a real-world ad dataset with rich auxiliary attributes. © 2021, CC BY-NC-SA.

关键词： Marketing

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Prioritizing Strategies for Building the Resilience of Public Health Systems to Disasters Across Multiple Communities and Countries

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International Journal of Disaster Risk science 2024年第1期15卷 1-17页

作者： Benjamin J.Ryan Mayumi Kako Shelby Garner Rok Fink Ismail Tayfur Jonathan Abrahams Sanjaya Bhatia Adriana Campelo Matthew Fendt Alicia Fontenot Nahuel Arenas Garcia Tim Hatch Ryoma Kayano LaShonda Malrey‑Horne Makiko MacDermot Md Moshiur Rahman Chaverle Noel Shuhei Nomura Jeremy P.Novak Maria Opazo Kendell Oliver Luciana Peters Sohel Rahman PerihanŞimşek Andrew Stricklin Raymond Swienton Bryan W.Brooks Frist College of Medicine Belmont UniversityNashvilleTN 37212USA Department of Environmental Science Environmental Health Science ProgramCollege of Arts and SciencesBaylor UniversityWacoTX 76798USA Division of Nursing Science School of Biomedical and Health SciencesHiroshima UniversityHiroshima 734-8553Japan Gordon E.Inman College of Health Sciences and Nursing Belmont UniversityNashvilleTN 37212USA Faculty of Health Sciences University of Ljubljana1000 LjubljanaSlovenia Department of Emergency Medicine University of Health Sciences34668 IstanbulTurkey Monash University Accident Research Center MelbourneVIC 3800Australia Global Education and Training Institute United Nations Ofce for Disaster Risk ReductionIncheon 22004Republic of Korea Regional Ofce for the Americas and Caribbean United Nations Ofce for Disaster Risk ReductionPanama City 0843-03441Panama Engineering and Computer Science Baylor UniversityWacoTX 76798USA Department of Public Health Robbins College of Health and Human SciencesBaylor UniversityWacoTX 76798USA Alabama Department of Public Health MontgomeryAL 36092USA World Health Organization Centre for Health Development(WHO Kobe Centre) Kobe 651-0073Japan Waco-McLennan County Public Health District WacoTX 76707USA Department of Health Policy and Management School of MedicineKeio UniversityTokyo 160-8582Japan Department of Global Health Policy Graduate School of MedicineThe University of TokyoTokyo 113-0033Japan Tokyo Foundation for Policy Research Tokyo 106-6234Japan Global Development College BrisbaneQLD 4000Australia Municipality of Pudahuel 13124 PudahuelChile City Hall of Porto Alegre Porto Alegre 90010-170Brazil Ministry of Health and Family Welfare Government of the People’s Republic of BangladeshDhaka 1000Bangladesh Department of Emergency Aid and Disaster Management Trabzon University61335 TrabzonTurkey Department of Emergency Medicine UT SouthwesternDallasTX 75390USA

The COVID-19 pandemic highlighted the urgent need to strengthen public health *** response,the United Nations Disaster Risk Reduction(UNDRR) Public Health System Resilience Scorecard(Scorecard) was applied in workshops across multiple *** aim of our research was to explore the workshop findings to develop priority strategies for strengthening public health system *** conducted a workshop from 14 to 16 March 2023,at the UNDRR Global Education and Training Institute in Incheon,Republic of Korea.A sequential modified Delphi method was utilized to develop a set of prioritized resilience *** were drawn from 70 strategies identified from 13 distinct workshops in eight *** two surveys,23 strategies were *** received ratings of "High" or Very High" from89% of *** related to the inclusion of public health risks in emergency plans,integrating multidisciplinary teams into public health,enabling local transport mechanisms,and improving the ability to manage an influx of *** Scorecard provides an adaptable framework to identify and prioritize strategies for strengthening public health system *** leveraging this methodology,our study demonstrated how resilience strategies could inform disaster risk reduction funding,policies,and actions.

关键词： Health system resilience Public policy Resilience scorecard Workshops

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Ginkgolide B facilitates muscle regeneration via rejuvenating osteocalcin-mediated bone-to-muscle modulation in aged mice

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Journal of cachexia, sarcopenia and muscle 2023年第3期14卷 1349-1364页

作者： Belle Yu-Hsuan Wang Yi-Fan Chen Allen Wei-Ting Hsiao Wan-Jing Chen Chien-Wei Lee Oscar Kuang-Sheng Lee Musculoskeletal Research Laboratory SH Ho Scoliosis Research Laboratory Department of Orthopaedics and Traumatology Faculty of Medicine The Chinese University of Hong Kong Hong Kong. Center for Neuromusculoskeletal Restorative Medicine CUHK InnoHK Centres Hong Kong Science Park Hong Kong. The Ph.D. Program for Translational Medicine College of Medical Science and Technology Taipei Medical University Taipei Taiwan. Graduate Institute of Translational Medicine College of Medical Science and Technology Taipei Medical University Taipei Taiwan. International Ph.D. Program for Translational Science College of Medical Science and Technology Taipei Medical University Taipei Taiwan. Master Program in Clinical Genomics and Proteomics School of Pharmacy Taipei Medical University Taipei Taiwan. Department of Orthopaedics and Traumatology Faculty of Medicine Prince of Wales Hospital The Chinese University of Hong Kong Hong Kong. Center for Translational Genomics & Regenerative Medicine Research China Medical University Hospital China Medical University Taichung Taiwan. Department of Biomedical Engineering China Medical University Taichung Taiwan. Department of Orthopedics China Medical University Hospital Taichung Taiwan. Institute of Clinical Medicine National Yang Ming Chiao Tung University Taipei Taiwan.

BACKGROUND:The progressive deterioration of tissue-tissue crosstalk with aging causes a striking impairment of tissue homeostasis and functionality, particularly in the musculoskeletal system. Rejuvenation of the systemic and local milieu via interventions such as heterochronic parabiosis and exercise has been reported to improve musculoskeletal homeostasis in aged organisms. We have shown that Ginkgolide B (GB), a small molecule from Ginkgo biloba, improves bone homeostasis in aged mice by restoring local and systemic communication, implying a potential for maintaining skeletal muscle homeostasis and enhancing regeneration. In this study, we investigated the therapeutic efficacy of GB on skeletal muscle regeneration in aged mice. METHODS:Muscle injury models were established by barium chloride induction into the hind limb of 20-month-old mice (aged mice) and into C2C12-derived myotubes. Therapeutic efficacy of daily administrated GB (12 mg/kg body weight) and osteocalcin (50 μg/kg body weight) on muscle regeneration was assessed by histochemical staining, gene expression, flow cytometry, ex vivo muscle function test and rotarod test. RNA sequencing was used to explore the mechanism of GB on muscle regeneration, with subsequent in vitro and in vivo experiments validating these findings. RESULTS:GB administration in aged mice improved muscle regeneration (muscle mass, P = 0.0374; myofiber number/field, P = 0.0001; centre nucleus, embryonic myosin heavy chain-positive myofiber area, P = 0.0144), facilitated the recovery of muscle contractile properties (tetanic force, P = 0.0002; twitch force, P = 0.0005) and exercise performance (rotarod performance, P = 0.002), and reduced muscular fibrosis (collagen deposition, P < 0.0001) and inflammation (macrophage infiltration, P = 0.03). GB reversed the aging-related decrease in the expression of osteocalcin (P < 0.0001), an osteoblast-specific hormone, to promote muscle regeneration. Exogenous osteocalcin supplementation was

关键词： Ginkgolide B aging osteocalcin rejuvenation skeletal muscle regeneration

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Chemically Binding Scaffolded Anodes with 3D Graphene Architectures Realizing Fast and Stable Lithium Storage

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Research 2019年第1期2019卷 662-670页

作者： Ping Wu Zhiwei Fang Anping Zhang Xiao Zhang Yawen Tang Yiming Zhou Guihua Yu Materials Science and Engineering Program and Department of Mechanical Engineering Te University of Texas at AustinAustinTexas 78712USA Jiangsu Key Laboratory of New Power Batteries Jiangsu Collaborative Innovation Center of Biomedical Functional MaterialsSchool of Chemistry and Materials ScienceNanjing Normal UniversityNanjing 210023China

Tree-dimensional(3D)graphene has emerged as an ideal platform to hybridize with electrochemically active materials for improved ***,for lithium storage,current anodic guests ofen exist in the form of nanoparticles,physically attached to graphene hosts,and therefore tend to detach from graphene matrices and aggregate into large congeries,causing considerable capacity fading upon repeated ***,we develop a facile double-network hydrogel-enabled methodology for chemically binding anodic scafolds with 3D graphene *** tin-based alloy anodes as an example,the doublenetwork hydrogel,containing interpenetrated cyano-bridged coordination polymer hydrogel and graphene oxide hydrogel,is directly converted to a physical-intertwined and chemical-bonded Sn−Ni alloy scafold and graphene architecture(Sn−Ni/G)dual *** unique dual framework structure,with remarkable structural stability and charge-transport capability,enables the Sn−Ni/G anode to exhibit long-term cyclic life(701 mA h g^(−1) afer 200 cycles at 0.1 A g^(−1))and high rate performance(497 and 390 mA h g^(−1) at 1 and 2 A g^(−1),respectively).Tis work provides a new perspective towards chemically binding scafolded low-cost electrode and electrocatalyst materials with 3D graphene architectures for boosting energy storage and conversion.

关键词： anodic attached boosting

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Elbow angle estimation for medical rehabilitation device based on EMG sensor with ARIMAX method

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AIP Conference Proceedings 2022年第1期2663卷

作者： Amelia Lita I. Made Mas Dwiyana Prasetya Wibawa Alinda Anggraini Akif Rahmatillah Riries Rulaningtyas Alfian Pramudita Putra Khusnul Ain 1Biomedical Engineering Study Program Department of Physics Faculty of Science and Technology Universitas Airlangga Surabaya 60115 Indonesia

In the development of medical rehabilitation tools, muscle signals or electromyographic (EMG) are often used as feedback from the body in order to facilitate normal movement patterns. Through this signal, the force, torque, and angle can be estimated to produce the desired movement in the robot to assist patients in rehabilitation therapy. There had been various research done on estimating the extremities' joint angle, including the system identification method with Auto-Regressive Integrated Moving Average with Exogenous Input (ARIMAX) model structure. This research aims to model biceps brachii's EMG signal and the elbow joint angle using the ARIMAX model structure. In modeling, the order of the model was selected using the methods of minimizing the AIC (Akaike Information Criteria) and RMSE (Root Mean Square Error) values. The reported results showed the comparison of the outcome and model orders from three data sets with a frequency sampling of 47.6690 Hz, l66.3248 Hz, and 935.6743 Hz, respectively. The frequency sampling of l66.3248 Hz results showed the best accuracy performance of the ARIMAX model structure. The model order which is selected based on the minimization of the RMSE value obtained the best estimate, while the order models were selected based on the minimization of the AIC value showed a more consistent performance on the data estimation and validation.

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