Development of Practical Wrist Rehabilitation Robot by Mirror Effect

The authors have developed wrist rehabilitation robot for hemiplegic patients. The robot is much effective for repetitive rehabilitation, and useful for patients. Mirror effect control system is developed based on the experience that a patient can easily move a limb while moving a limb on the other side. The effectiveness is successfully confirmed by the practical test in the hospital.3rd International Conference on Advanced Materials Design and Mechanics, ICAMDM 2014; ; 23 May 2014 through 24 May 201

Prototype of positioning system for automatic motion control of underwater robot

Recently, the needs of underwater robot used for many kinds of underwater work become higher and higher. In order to control the motion of underwater robot automatically, it is an indispensable to measure its position correctly in real time. Conventional systems are based on time difference or phase lag. However, they must use expensive components, such as transponders or atomic clock, and its system becomes complex. Additionally they require a lot of signal-processing time, so they cannot be used for motion control. As a result, there exist no system which can be used for automatic motion control of underwater robot. Since 2008, we have proposed a new positioning system based on sound propagation loss and sensor network. In this system, we set many buoys that install sound reception unit, GPS receiving equipment and sensor network system on the surface of water. We use sound propagation loss to calculate the distance between robot and buoys, and with combining distance and position data for each buoy, we estimate robot position. In former study, we found that we can get higher SN ratio of signal to measure distance with using the sound of multiple frequency, we proposed a new distance measurement method M_SPL. It was introduced on the paper of “Underwater Acoustics Positioning System Based on Propagation Loss and Sensor Network” (OCEANS 2012 KOREA). In this paper, we introduce the design and construction of prototype system for this positioning system as the results of latest study. It contains transmission unit, reception unit and center unit. Transmission unit follows with principle of M_SPL, is set into underwater robot used as sound source. We also complete reception unit to be used to receive sound signal and calculate distance automatically. Finally, center unit receive distance and each buoy position data what are sent from reception unit, position can be calculated by using these data. We confirmed the performance of transmission unit and r- ceiving unit in water-tank, and it showed that both units can be used effectively. Now we are on a final test of this system in open sea for the actual use of surveying sea floor with the underwater robot made by NAGASAKI UNIVERSITY in Japan.OCEANS\u2714 MTS/IEEE Conference on April 7-10, 2014, in Taipei, Taiwan

Development of a rehabilitative training robot for both wrists

The authors developed a rehabilitative training robot which detects and analyzes the intention when a person tries to move a wrist with a biological signal, such as muscle action potential, and makes a wrist movement as one\u27s intention. It consists of a grip for wrists, an actuator, a biological signal primary means, biological signal processing part, and a training controller. Due to its small, compact size, training can be performed in a small space at a hospital or home. The new system which can conduct both wrists training is successfully developed for practical uses.2013 7th ICME International Conference on Complex Medical Engineering, CME 2013; Beijing; China; 25 May 2013 through 28 May 201

Research and Development of Unmanned Aviation Vehicle System for Disaster Countermeasures

The authors developed low cost unmanned helicopter, real time video data transmitting system and a new type flying robots. The effectiveness is confirmed by real field experiment tests. Hazard map for disaster countermeasures is made on the basis of data obtained by the aviation vehicle system. The hazard map is delivered directly to a smart phone and a tablet PC. Therefore residents can keep track of disaster information quickly and accurately.2013 International Conference on Materials, Transportation and Environmental Engineering, CMTEE 2013; Taichung; Taiwan; 21 August 2013 through 23 August 201

Dnmt3a Regulates Proliferation of Muscle Satellite Cells via p57Kip2

<div><p>Cell differentiation status is defined by the gene expression profile, which is coordinately controlled by epigenetic mechanisms. Cell type-specific DNA methylation patterns are established by chromatin modifiers including <i>de novo</i> DNA methyltransferases, such as <i>Dnmt3a</i> and <i>Dnmt3b</i>. Since the discovery of the myogenic master gene <i>MyoD</i>, myogenic differentiation has been utilized as a model system to study tissue differentiation. Although knowledge about myogenic gene networks is accumulating, there is only a limited understanding of how DNA methylation controls the myogenic gene program. With an aim to elucidate the role of DNA methylation in muscle development and regeneration, we investigate the consequences of mutating <i>Dnmt3a</i> in muscle precursor cells in mice. <i>Pax3</i> promoter-driven <i>Dnmt3a</i>-conditional knockout (cKO) mice exhibit decreased organ mass in the skeletal muscles, and attenuated regeneration after cardiotoxin-induced muscle injury. In addition, <i>Dnmt3a</i>-null satellite cells (SCs) exhibit a striking loss of proliferation in culture. Transcriptome analysis reveals dysregulated expression of <i>p57Kip2</i>, a member of the Cip/Kip family of cyclin-dependent kinase inhibitors (CDKIs), in the <i>Dnmt3a</i>-KO SCs. Moreover, RNAi-mediated depletion of <i>p57Kip2</i> replenishes the proliferation activity of the SCs, thus establishing a role for the <i>Dnmt3a</i>-<i>p57Kip2</i> axis in the regulation of SC proliferation. Consistent with these findings, <i>Dnmt3a</i>-cKO muscles exhibit fewer Pax7⁺ SCs, which show increased expression of p57Kip2 protein. Thus, Dnmt3a is found to maintain muscle homeostasis by epigenetically regulating the proliferation of SCs through <i>p57Kip2</i>.</p></div

Loss of <i>Dnmt3a</i> leads to elevated expression of <i>p57Kip2</i>.

<p>(A) RT-qPCR analysis of <i>Dnmt3a</i> in Ax-Cre <i>Dnmt3a</i>-KO and WT SCs before differentiation. (B) A heat map showing the expression levels of cell cycle-regulating genes based on the transcriptome analysis of Ax-Cre <i>Dnmt3a</i>-KO and WT SCs. <i>p57Kip2</i> (<i>Cdkn1c</i>) is highly expressed in <i>Dnmt3a</i>-KO SCs. (C) RT-qPCR analysis of <i>p57Kip2</i> in <i>Pax7-Cre; Dnmt3a</i>-KO and WT SCs. (D) Representative photomicrographs of <i>Pax7-Cre; Dnmt3a</i>-KO and WT SCs stained with p57Kip2 and DAPI. Scale bar—30 μm. (E) Dot plots of mean signal intensities of fluorescence in each SC stained with an anti-p57Kip2 antibody; Gray bars represent mean ± SD, ***p<0.001, Student’s t-test.</p

<i>p57Kip2</i> is a methylation target of Dnmt3a and regulates proliferation of SCs.

<p>(A) A schematic diagram of bisulfite sequencing at the <i>p57Kip2</i> locus of <i>Dnmt3a</i>-KO and WT SCs on day 0 and day 2 after inducing differentiation. White circles and black circles indicate unmethylated and methylated CpG sites, respectively. The transcription start site (TSS) is indicated by the bent arrow. The gray bar represents the sequencing target (+1 corresponds to the TSS). (B) Average methylation frequency of CpG sites at the <i>p57Kip2</i> locus; *p<0.05, Student’s t-test. SEMs between sequences are shown. (C) <i>p57Kip2</i> knockdown by siRNA in <i>Dnmt3a</i>-KO and WT SCs; *p<0.01 Student’s t-test. (D) Cell proliferation assay after <i>p57Kip2</i> knockdown. Numbers of SCs 3 days after siRNA transfection are shown. Reduced cell proliferation in <i>Dnmt3a</i>-KO SCs is partially rescued by <i>p57Kip2</i> knockdown; *p<0.05 Student’s t-test. Data represent mean ± SEM. (E) Quantification of PHH3 positive SCs with p57Kip2 knock-down: *p<0.05 Student’s t-test. Data represent mean ± SEM. The sequences of all primers used for PCR are listed in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1006167#pgen.1006167.s012" target="_blank">S1 Table</a>.</p

A schematic diagram of the <i>Dnmt3a-p57Kip2</i> axis in muscle satellite cells.

<p><i>Dnmt3a</i> regulates proliferation of SCs via DNA methylation of the <i>p57Kip2</i> promoter. Loss of <i>Dnmt3a</i> leads to decreased proliferation of SCs, resulting in reduced muscle mass and impaired muscle regeneration.</p

Loss of <i>Dnmt3a</i> leads to impaired proliferation of muscle satellite cells.

<p>(A) A schematic diagram of conditional <i>Dnmt3a</i>-KO in muscle SCs. Triangles represent loxP sites. (B) Genomic <i>Dnmt3a</i> levels in the SCs; <i>Col2a1</i> promoter served as the control; ***p<0.001, Student’s t-test. (C) <i>Dnmt3a</i> mRNA levels in the SCs; <i>Gapdh</i> served as the control. <i>Dnmt3a</i> expression in cKO SCs is below detectable levels. (D) Representative phase-contrast microscopic images of <i>Dnmt3a</i>-KO and WT SCs. Both KO and WT cells were disseminated at the same cell density on Day 0. Scale bar—200 μm. (E) <i>In vitro</i> cell proliferation assay shows significantly reduced proliferation of <i>Dnmt3a</i>-KO SCs compared to WT SCs; ***p<0.001, two-way repeated measures ANOVA. (F) Representative images of PHH3 immunocytochemistry of <i>Dnmt3a</i>-KO and WT SCs. Arrowheads indicate PHH3⁺ cells. Scale bar—30 μm. (G) Quantification of PHH3⁺ cells in <i>Dnmt3a</i>-KO and WT SCs; ***p<0.001, Student’s t-test. Data represent mean ± SEM.</p

Loss of <i>Dnmt3a</i> causes reduced muscle mass in mice.

<p>(A) A schematic diagram of muscle precursor cell-specific <i>Dnmt3a</i> deletion by Cre-loxP recombination. Triangles represent loxP sites. (B) Genomic DNA levels of <i>Dnmt3a</i> in tibialis anterior muscles; <i>Col2a1</i> promoter served as the control; *p<0.05, Student’s t-test. (C) RT-qPCR analysis for <i>Dnmt3a</i> using muscle tissues; <i>Gapdh</i> served as the control; *p<0.05, Student’s t-test. (D) <i>Dnmt3a</i>-cKO mice have smaller body sizes than WT controls. (E) <i>Dnmt3a</i>-cKO mice weigh less than WT controls; *p<0.05, **p<0.01, ***p<0.001, Student’s t-test. (F) X-ray whole body imaging of <i>Dnmt3a</i>-cKO and WT mice; (G, H) Muscle mass in male (G) and female (H) mice. (Left) CT images of the lower hindlimb of <i>Dnmt3a</i>-cKO and WT mice. CT images of the slices that have the maximum muscle CSA in each limb are shown. (Right) The ratio of maximum CSA of the muscle divided by that of the bone at the same slice level demonstrates disproportionately reduced muscle mass in <i>Dnmt3a</i>-cKO mice; *p<0.05, **p<0.01, Student’s t-test. (I) HE staining of the gastrocnemius muscle cross sections of <i>Dnmt3a</i>-cKO and WT mice. (J) Distribution of myofiber CSAs in <i>Dnmt3a</i>-cKO and WT muscles. (K) Box plots for myofiber CSAs in <i>Dnmt3a</i>-cKO and WT muscles; ***p<0.001, Mann-Whitney U test. Data represent mean ± SEM. Hm—<i>Dnmt3a</i> homozygous knockout; Ht—<i>Dnmt3a</i> heterozygous knockout. Scale bar—200 μm.</p