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Many hemiplegic patients, especially those having paralysis at hand, need continuous rehabilitation after leaving hospitals. To support rehabilitation at home, we have developed a robotic device which provides automated rehabilitation and evaluation of hand motor function recovery. The device monitors the patient's finger movements during voluntary finger tasks. Our idea is to calculate the extent of recovery by asking a patient to perform a specific voluntary movement (task) and calculating the dissimilarity between a typical healthy person's movement and that measured by this device. In our previous studies, we only clarified the evaluation methods for a single task. This paper proposes the integration of these values into a single measurement value by using binary logistic regression. This integration provides the patient with a probability of being classified as a patient and a healthy subject. A home user should easily understand the value. The experimental results show that the integrated evaluation has higher performance over the single task evaluation.

Continuous hand rehabilitation after discharge is important for hemiplegic patients to regain an independent finger movement. However, most patients cannot rehabilitate by themselves without therapists. For this problem, robotic rehabilitation has been investigated to support patients even at home. Most of the programs performed by these robots are focusing on the assistance for voluntary movement. However, the approach to the voluntary movement is not enough for regaining dexterous movement. Voluntary suppression of body parts that should not move is important. However, previous studies focusing on voluntary suppression are few. In this paper, we show a detailed program for voluntary suppression rehabilitation. The program is performed by our robotic finger rehabilitation device aiming at home use. In this program, a patient is requested to flex and extend an index finger independently. During moving, individual pressure sensors monitor the other fingers. If the device detects unnecessary movements such as compensatory movement at some fingers, the patient is notified that unnecessary movements are found there. The detection is based on $3\sigma$ range of healthy subject’s finger pressure data which was constructed by using Gaussian Process Regression. Through experiments with hemiplegic patients, we have shown that the frequency of deviation of patients’ data from $3\sigma$ range of healthy subjects decreases according to the degree of recovery.

Home rehabilitation is much in need in ageing societies. Especially, for hemiplegia patients who have paralysis at fingers, it is known that long continuous rehabilitation is effective for recovery. To automate home rehabilitation without the help of a medical specialist, it is desirable not only to provide a rehabilitation procedure but also to give the condition of the paralysis of the patient. In our previous studies, we proposed a robotic device to foster separative voluntary finger lift movement. The device mounts pressure sensors at each finger to monitor the degree of unwanted finger movements. However, it is not clear which is an effective way to measure the condition of paralysis by these finger pressure time series data. In this paper, we propose a new measurement method that is based on the “coordinated movement” of fingers. A patient is asked to perform 4 tasks: pinch2 (pinch movement by index finger and thumb), pinch3 (pinch movement by index, middle and thumb), grasp2 (grasp movement by the ring and little finger), and grasp3 (grasp movement by the middle, ring, and little finger). All pressure time series of finger movements are quantified their dissimilarities with data of standard healthy subjects. We found that the grasp3 has the largest correspondence relation with the degree of paralysis.

doi: 10.18178/ijmerr.5.4.288-294, As the number of patients suffering from cerebrovascular disease (CVD) increases, it is desired that the patients will be able to receive proper rehabilitation at home without specialists' help. For this, we have developed a finger-dedicated robotic rehabilitation support device that is compact enough to be used at home. The device induces voluntary muscular contraction at a finger by the help of power assisted finger lifter. The lifter is a simple lever on which the patient places his/her finger. Although the mechanism is simple to be used at home, there is a question whether applying only a finger-dedicated rehabilitation is sufficient to improve the quality of life of a patient including recovery of upper limb functions. To answer this question, we conducted experiments that show skill transfer from finger to elbow. In the experiments, subjects are asked to train to produce a specified force at their fingers. After training, it was shown that the subjects are able to perform the same skill by using elbow, where no training was performed. The results support that the finger-dedicated rehabilitation would be appropriate as a target for a robotic home rehabilitation device.

Paralysis of fingers, which is caused by Hemiplegia, is difficult to recover. Patients often forced to leave hospital with paralysis remaining at hand. By this, a continuous rehabilitation at home is needed. However, it is difficult to carry out finger rehabilitation without help of therapists. To this end, we have been proposing an automated finger rehabilitation device which realizes home rehabilitation. A patient is asked by device to lift a finger, and the device measures whether undesirable movements are found on the other fingers by pressure sensors. To monitor an involuntary movement, it is necessary to evaluate the degree of the patient's condition of recovery. For this, we proposed a quantification method in our previous study. The method is based on the hypothesis that a patient is regarded as making recovery if his/her movement gets close to that of a healthy person. However, we consider only four fingers (index, middle, ring, little) are used to evaluate the degree of recovery because the thumb is different from the other finger in an anatomical structure. In this paper, we show a new recovery evaluation method that involves the sensor signals of all 5 fingers. We explain two possible evaluation methods: one is the model less simple integration method, and another is an integration by Generalized Linear Model (GLM). Comparing these methods, we conclude that the integration method by GLM provides a good scalar measurement of recovery, which was validated by the experiments conducted with patients who were previously evaluated by clinical scale.

指導教員：三上 貞芳

Plant associations by bradyrhizobia have been detected not only in leguminous plants, but also in non-leguminous species including rice. Bradyrhizobium sp. SUTN9-2 was isolated from Aeschynomene americana L., which is a leguminous weed found in the rice fields of Thailand. This strain promoted the highest total rice (Oryza sativa L. cultivar Pathum Thani 1) dry weight among the endophytic bradyrhizobial strains tested, and was, thus, employed for the further characterization of rice-Bradyrhizobium interactions. Some known bacterial genes involved in bacteria-plant interactions were selected. The expression of the type III secretion component (rhcJ), type IV secretion component (virD4), and pectinesterase (peces) genes of the bacterium were up-regulated when the rice root exudate was added to the culture. When SUTN9-2 was inoculated into rice seedlings, the peces, rhcJ, virD4, and exopolysaccharide production (fliP) genes were strongly expressed in the bacterium 6-24 h after the inoculation. The gene for glutathione-S-transferase (gst) was slightly expressed 12 h after the inoculation. In order to determine whether type III secretion system (T3SS) is involved in bradyrhizobial infections in rice plants, wild-type SUTN9-2 and T3SS mutant strains were inoculated into the original host plant (A. americana) and a rice plant (cultivar Pathum Thani 1). The ability of T3SS mutants to invade rice tissues was weaker than that of the wild-type strain; however, their phenotypes in A. americana were not changed by T3SS mutations. These results suggest that T3SS is one of the important determinants modulating rice infection; however, type IV secretion system and peces may also be responsible for the early steps of rice infection.

In current health care system, it is difficult to have sufficiently long period of rehabilitation in hospital. Therefore, an automated rehabilitation support at home is needed. To this end, we developed a simple robotic device that will support index finger rehabilitation for patients who suffer from aftereffects of hemiplegia. The purpose of the device is to foster voluntary movement of index finger. It has pressure sensors for all fingers, and a power assisted index finger lift mechanism. The power assist lift supports to lift the patient's finger if the lift by voluntary movement is insufficient. The pressure sensors are used to monitor the simultaneous movement of all fingers. By this, the system checks if undesirable movements such as synergetic movements are found. The information is integrated to provide a quantitative measure of recovery. Patients are able to monitor his/her condition and continue rehabilitation as needed. Our device is simple and compact so that it should be placed on a desk at home. The device is used by placing the patient's hand on keyboards. This allows the patients to see his/her fingers during rehabilitation and is expected to enhance the effect of rehabilitation by seeing the result of voluntary movement at his/her real fingers. This paper shows these concepts and the hardware design of this device in detail. This paper also discusses the results of the measurement of hemiplegia patients and the healthy subjects by this device. The results show that the explicit difference was seen between the patients and the healthy subjects, which shows the effectiveness of this design.

After leaving hospital, patients can carry out rehabilitation by using rehabilitation devices. However, they cannot evaluate the recovery by themselves. For this problem, a device which can both carry out the rehabilitation and evaluation of the degree of recovery is required. This paper proposes the method that quantifies the recovery of the paralysis of fingers to evaluate a patient automatically. A finger movement is measured by a pressure sensor on the rehabilitation device we have developed. A measured data is used as a time-series signal, and the recovery of the paralysis is quantified by calculating the dissimilarity between a healthy subject's signal and the patient's signal. The results of those dissimilarities are integrated over all finger to be used as a quantitative scale of recovery. From the experiment conducted with hemiplegia patients and healthy subjects, we could trace the process of the recovery by the proposed method.

X-ray structure determination and deuteration of nattokinase were performed to facilitate neutron crystallographic analysis., Nattokinase (NK) is a strong fibrinolytic enzyme, which is produced in abundance by Bacillus subtilis natto. Although NK is a member of the subtilisin family, it displays different substrate specificity when compared with other subtilisins. The results of molecular simulations predict that hydrogen arrangements around Ser221 at the active site probably account for the substrate specificity of NK. Therefore, neutron crystallographic analysis should provide valuable information that reveals the enzymatic mechanism of NK. In this report, the X-ray structure of the non-hydrogen form of undeuterated NK was determined, and the preparation of deuterated NK was successfully achieved. The non-hydrogen NK structure was determined at 1.74 Å resolution. The three-dimensional structures of NK and subtilisin E from Bacillus subtilis DB104 are near identical. Deuteration of NK was carried out by cultivating Bacillus subtilis natto in deuterated medium. The D2O resistant strain of Bacillus subtilis natto was obtained by successive cultivation rounds, in which the concentration of D2O in the medium was gradually increased. NK was purified from the culture medium and its activity was confirmed by the fibrin plate method. The results lay the framework for neutron protein crystallography analysis.

Mass spectrometry (MS) is a method of analyzing ions based on their mass/charge (m/z) ratios. The m/z peak identification requires speculation on the ionic unit-charge states. This problem can be solved by using superconducting junction devices to measure the kinetic energies of single molecules. However, the kinetic energy measurement is followed by the dead time of 1–20 µs, which is fatally slow for modern high-resolution time-of-flight (TOF) analyzers. In this paper, we demonstrate that a superconducting nano-stripline detector (SSLD) composed of a 10-nm-thick and 800-nm-wide NbN strip realizes the charge-state derivation, and furthermore satisfies the ideal MS detector specifications such as a nano-second response, a short recovery time, a wide mass range, and no noise. Copyright © 2010 John Wiley & Sons, Ltd.

The polymerization of δ-valerolactone (δ-VL) using 3-phenyl-1-propanol (3-Ph-PrOH) as the initiator and trifluoromethanesulfonimide (HNTf2) as the catalyst in CH2Cl2 at 27 °C with the [δ-VL]0/[3-Ph-PrOH]0/[HNTf2]0 ratio of 100:1:0.1 proceeded homogeneously to afford a poly(δ-valerolactone) (PVL) with a narrow polydispersity index. The molecular weight determined from the 1H NMR analysis, Mn = 9600 g mol−1, fairly agreed with that estimated from the initial ratio of [δ-VL]0/[3-Ph-PrOH]0, Mn,theo = 9400 g mol−1. In addition, the kinetic and chain extension experiments confirmed that the HNTf2-catalyzed ROP proceeded in a living fashion. The 1H NMR, SEC, and MALDI-TOF MS measurements of the obtained PVL clearly indicated the presence of the initiator residue at the chain end, showing that the HNTf2-catalyzed ROP of δ-VL proceeded through a living mechanism. The HNTf2-catalyzed ROP of δ-VL with functional initiators, such as 6-azido-1-hexanol, 2,3,4,5,6-pentafluorobenzyl alcohol, and N-(2-hydroxyethyl)maleimi...

Super water- and highly oil-repellent films have been synthesized by electrochemical polymerization of some new monomers of fluorinated alkylpyrroles. The surface roughness of the films was controlled by the polymerization conditions such as concentration of monomers, applied voltages, polymerization time and so on. The contact angles on the most liquid-repellent film surfaces are about 160° and 140° for water and salad oil, respectively. The sliding angle for the salad oil is 9.5 ± 2.5° which is much better than that on the poly(alkylpyrrole) films coated with a fluorinated alkylsilane coupling agent. Thermal and organic solvent stability for the liquid-repellent property of the films were quite good to be used practically.

The Bronsted acid, trifluoromethanesulfonimide (HNTf2), was revealed to act as a highly efficient activator for group transfer polymerization (GTP) of methyl methacrylate (MMA) at ambient temperature. The HNTf2-catalyzed GTP of MMA initiated by 1-methoxy-1-trimethylsilyloxy-2-methyl-propene (MTS) proceeded in a living manner to produce poly(methyl methacrylate)s (PMMA) with controlled molecular weights and narrow molecular weight distributions. Kinetic measurement and postpolymerization experiment further supported that the polymerization proceeded in a living fashion. Furthermore, from the MALDI-TOF MS analysis, only one series of peaks was observed, which was in perfect agreement with the molecular weight of PMMA possessing the MTS initiator residue and the desilylated chain end, showing that the HNTf2-catalyzed GTP of MMA proceeded without any side reactions. The syndiotacticity of the PMMA obtained by the HNTf2-catalyzed GTP increased with the decreasing polymerization temperature; the syndiotactic (r...

Conventional time-of-flight mass spectrometers equipped with microchannel plate (MCP) detectors give only m/z values, which remains ambiguous over charge number (z). In contrast, the matrix-assisted laser desorption/ionization (MALDI) TOF mass spectrometer (TOF-MS) with a superconducting tunnel junction (STJ) detector—Super-TOF-MS—enables kinetic energy measurement with mass-independent detection efficiency. Therefore, since kinetic energy is proportional to z, it is possible to determine mass (m) unambiguously.The Super-TOF-MS is effective for analysing high-mass fragments in immunoglobulin solution. Specifically, unknown peaks appearing on mass spectra for immunoglobulin G (IgG) samples can be identified as IgG fragments using kinetic energy measurements and the mass-independent detection efficiency of the STJ detector. A complete IgG molecule has two heavy and two light chains that are connected by disulfide bonds. The heavy and light chains include four and two structural domains, respectively (12 domains in total). The charge-state discrimination mass analysis enables the assignment of m values to the unknown peaks. The existence of pairs for which the m-value sum is equal to that of the intact IgG reveals that there are several fragment types consisting of different numbers of structural domain units.

Time-of-flight mass spectrometry (TOF-MS) with matrix assisted laser desorption/ionization (MALDI) has become an essential spectrometry for a research field of bioscience. However, the mass limit of the conventional TOF-MS spectrometer frequently prevent us from measuring the mass values for large biomolecules without chemical or enzymatic digestion. Overcoming the circumstance, a TOF-MS instrument with superconducting tunnel junction (STJ) detectors (Super TOF-MS) were developed. The direct observation of the kinetic energy for each ion by Super TOF-MS leads to the two considerable capabilities: the observation of large proteins up to 1 MDa and the ionic charge-state discrimination of them, which are impossible with conventional ion detectors. In this paper, it is demonstrated that the these advantages of Super TOF-MS is a powerful tool at performing a fragment analysis of, for example, immunoglobulin G (IgG) which is a large protein playing a key role in an immune system.

We have conducted a study to compare various preparation methods, including a certain number of new methods, to find which ones are best suited to the morphological evaluation of nerve fibers in the pyramidal tract of the human medulla oblongata. Our main concern was to find fixation and staining methods that would minimize errors, especially regarding the tissue shrinkage ratio and the ease of staining. From the two fixation methods we examined, the most satisfactory was the secondary chromic acid fixation (which gave the best overall results when followed by nitrocellulose embedding), as it gives the lowest shrinkage ratio with the narrowest range (10 +/- 0%). Among the ten staining methods we tested, we found that the most suitable for morphological evaluation were the discriminative staining methods (Luxol fast blue-Periodic acid-Schiff-Hematoxylin stain, Masson-Goldner-Goto method and modified Hematoxylin-Eosin stain) and the silver impregnation methods (Luxol fast blue-Silver impregnation and Luxol fast blue-Silver impregnation-Periodic acid-Schiff-Hematoxylin).

To address the question as to how small-angle scattering is effectively applied to the cell, i.e., ahierarchically ordered system comprising multi-components of macro and small molecules, the size ofwhich ranges from 100 μm to several μm, we reconstructed SANS-J (pinhole small-angle neutronscattering spectrometer at research reactor JRR3, Tokai) to focusing & polarized neutron small-anglespectrometer (SANS-J-II), by employing focusing neutron lenses and high resolution photomultiplier.Consequently, an accessible minimum wave number qmin was improved from 3×10-3 A-1 to mediumultra-small angle scattering of 3×10-4 A-1. The focusing USANS method, thus developed, is crucialto fill the gap in wave number q between those covered by a double crystal method and by a conventionalpin-hole method.

Extra-slow-growing bradyrhizobia from root nodules of field-grown soybeans harbor abundant insertion sequences (ISs) and are termed highly reiterated sequence-possessing (HRS) strains. We analyzed the genome organization of HRS strains with the focus on IS distribution and symbiosis island structure. Using pulsed-field gel electrophoresis, we consistently detected several plasmids (0.07 to 0.4 Mb) in the HRS strains (NK5, NK6, USDA135, 2281, USDA123, and T2), whereas no plasmids were detected in the non-HRS strain USDA110. The chromosomes of the six HRS strains (9.7 to 10.7 Mb) were larger than that of USDA110 (9.1 Mb). Using MiSeq sequences of 6 HRS and 17 non-HRS strains mapped to the USDA110 genome, we found that the copy numbers of ISRj1, ISRj2, ISFK1, IS 1632 , ISB27, ISBj8, and IS 1631 were markedly higher in HRS strains. Whole-genome sequencing showed that the HRS strain NK6 had four small plasmids (136 to 212 kb) and a large chromosome (9,780 kb). Strong colinearity was found between 7.4-Mb core regions of the NK6 and USDA110 chromosomes. USDA110 symbiosis islands corresponded mainly to five small regions (S1 to S5) within two variable regions, V1 (0.8 Mb) and V2 (1.6 Mb), of the NK6 chromosome. The USDA110 nif gene cluster ( nifDKENXSBZHQW-fixBCX ) was split into two regions, S2 and S3, where ISRj1-mediated rearrangement occurred between nifS and nifB . ISs were also scattered in NK6 core regions, and ISRj1 insertion often disrupted some genes important for survival and environmental responses. These results suggest that HRS strains of soybean bradyrhizobia were subjected to IS-mediated symbiosis island shuffling and core genome degradation.

Although phytoremediation is a promising method for the removal of cadmium (Cd) from contaminated soil, plants remove Cd from soil slowly. To increase Cd uptake by plants, the effects of the application of chloride ions (Cl−) and a decrease in soil pH on the amount of Cd taken up by plants were investigated in a pot experiment using soil collected from paddy fields contaminated with Cd from mining wastewater. Three treatments were applied (all quantities were expressed on a 350 g soil basis): control (0.6 g (NH4)2SO4 as N fertilizer), Cl− application (0.5 g NH4Cl as N fertilizer and 0.15 g CaCl2) and low pH (0.8 g powdered sulfur plus NH4Cl and CaCl2). Sunflower (Helianthus annuus), kenaf (Hibiscus cannabinus) and sorghum (Sorghum vulgare) plants were grown in the treated soil for 33 days in a growth chamber under a 26°C/24°C day/night (16 h/8 h) regime. At harvest, the plants were separated into roots, stems and leaves, and the Cd concentration was determined. The application of Cl− approximatel...

To clarify the genetic basis to the diversity of denitrifying ability in soybean bradyrhizobia, we compared the end-products of denitrification (N2, N2O and NO2 � ) with the existence of denitrifying genes (napA, nirK, norCB and nosZ) of sixty phylogenetically diverse strains of Bradyrhizobium japonicum and B. elkanii. The results indicate that the existence of denitrifying genes directly determines phenotype (end-products) in most strains of B. japonicum. The denitrifying capability and gene set were reflected by phylogenetic position based on repeated sequences (RS)-fingerprints and 16S rRNA gene sequences. However, the denitrifying genes in HRS (highly repeated sequence-possessing) strains of B. japonicum, which were identified based on RS-fingerprints as having heavy hybridization, resulted in an inconsistent correlation probably because of genomic rearrangements. The evolutionary and ecological implications of the denitrifying genes and capability in soybean bradyrhizobia are discussed.

To evaluate the denitrification abilities of many Bradyrhizobium field isolates, we developed a new 15 N-labeled N 2 detection methodology, which is free from interference from atmospheric N 2 contamination. 30 N 2 ( 15 N 15 N) and 29 N 2 ( 15 N 14 N) were detected as an apparent peak by a gas chromatograph equipped with a thermal conductivity detector with N 2 gas having natural abundance of 15 N (0.366 atom%) as a carrier gas. The detection limit was 0.04% 30 N 2 , and the linearity extended at least to 40% 30 N 2 . When Bradyrhizobium japonicum USDA110 was grown in cultures anaerobically with 15 NO 3 − , denitrification product ( 30 N 2 ) was detected stoichiometrically. A total of 65 isolates of soybean bradyrhizobia from two field sites in Japan were assayed by this method. The denitrification abilities were partly correlated with filed sites, Bradyrhizobium species, and the hup genotype.

Using a preparation method composed of secondary chromic acid fixation, nitrocellulose embedding and Luxol fast blue-PAS-hematoxylin stain (a discriminative staining method) for the purpose of axonal discrimination, we examined 43 human spinal cords (31 males and 12 females) at the C5 level to find the differences due to gender and aging in axons of the lateral corticospinal tract. These results can be of great help in understanding the motor functions in relation to gender differences and the aging process.

弱酸性領域におけるアミノ多糖類キチンのアセチルアミノ基のプロトン化に基づく陰イオン交換体としての機能を利用し,銅をそのバソクプロインジスルホン酸(BCS)錯陰イオンとして濃縮した後,キチン相の反射吸光度を直接測定して銅を定量する方法を検討した.銅はpH5の水溶液20cm3からキチン相20mg上にBCSキレートとして短時間のかくはん時間で迅速に濃縮された.キチン相中の1μgまでの銅と484nmにおける固相吸光度との間に直線関係があった.銅0.5μg,9回測定の相対標準偏差は2.89%であった.VO3-やFe3+の許容量はやや低かったが,金属イオン,還元剤及び一般の無機陰イオンなどは1000～10000倍共存しても影響しなかった.本法により国立環境研究所の生物標準試料中の銅の定量を行った結果,鉄の共存量が比較的少ない試料中の銅の分析値は保証値と良く一致した.