A two-degree-of-freedom motor-powered gait orthosis for spinal cord injury patients

A number of orthoses have been developed to restore stance and walking in paraplegic subjects. Compliance, however, has been limited, mainly owing to walking effort. Use of the forces produced by actuators is an effective way to solve the problem of the considerable effort required for orthotic gait, namely high muscular effort and high energy expenditure. The purpose of the present study was to investigate the effects of assistance by external actuators on the orthotic gait of spinal cord injury (SCI) patients. Two kinds of linear actuator were developed by using direct current (d.c.) motors for assisting the knee and hip joint of a gait orthosis. They were mounted on the knee and hip joint of a commercial advanced reciprocating gait orthosis (ARGO), and a new two-degree-of-freedom externally powered gait orthosis was thus developed. The orthosis was assessed through inter-subject experiments on five male adult complete SCI patients. Owing to the short training period available for the assisted gait, simultaneous operation of both joint actuators was not conducted: either the knee actuation or the hip actuation was executed only. Thus, the knee actuator and the hip actuator were assessed with a T12 subject and with subjects for T5, T8, T11, and T12 respectively. The motions of the gaits, assisted by the linear actuators, were measured by a Vicon 370 system, and the general gait parameters and compensatory motions were evaluated. Results demonstrated that (a) all subjects could walk without falling, assisted either by the knee or the hip actuator; (b) both the knee and hip joint actuator increased the gait speed and the step length; (c) the knee flexion produced by the orthosis improved the dynamic cosmesis of walking; and (d) lateral compensatory motions as well as vertical ones tended to decrease when the hip joint was assisted, which could contribute to a reduction in walking effort.     

Macroscopic and microscopic changes in the surface layers of annealed and case-hardened steel rollers due to rolling contact

Measurements of the residual stress and observations of the microstructure at the surface and in the subsurface of rollers were performed during rolling contact fatigue tests of annealed 0.45% carbon steel and case-hardened nickel-chromium steel rollers. Compressive residual stresses in annealed rollers were induced by the rolling contact. With case-hardened rollers they were induced by heat treatment prior to the rolling contact fatigue tests. After the rolling contact fatigue tests the compressive residual stresses on the surface of the annealed rollers and in the subsurface of the case-hardened rollers relaxed; a characteristic substructure was formed by the stress cycles, which caused surface failure. It was confirmed that the microcracks leading to surface failure initiate on the surfaces of annealed rollers and in the subsurface of case-hardened rollers.     

Permeation measurements for investigating atomic hydrogen flux and wall pumping/fuelling dynamics in QUEST

In order to investigate the overall atomic hydrogen background and the dynamic characteristics of wall pumping/fuelling phenomenon, a permeation probe system has been developed and applied in the spherical tokamak QUEST. Reliability of measurements, within ±3% accuracy and a positive correlation with the hydrogen line emission over three orders of magnitude have been demonstrated for more than 3000 various plasma discharges. By comparison of the experimental permeation (flux) curves with the numerically simulated curves, the net incident atomic hydrogen flux is evaluated in the range of 1 × 10¹⁹ H m^?2 s^?1 to 4 × 10²⁰ H m^?2 s^?1. The atomic flux has been investigated as a function of various plasma operation parameters like RF power, gas pressure and magnetic configuration. Using the static particle balance and permeation measurements, the progress in wall conditioning has been investigated. An inverse correlation between the atomic hydrogen flux and improvement in wall pumping has been observed over the two campaigns.     

The Detection of Immunity against WT1 and SMAD4P130L of EpCAM+ Cancer Cells in Malignant Pleural Effusion

Malignant pleural effusion (MPE) provides a liquid tumor microenvironment model that includes cancer cells and immune cells. However, the characteristics of tumor antigen-specific CD8⁺ T cells have not been investigated in detail. Here, we analyzed MPE samples taken from a patient with pancreatic cancer who received a dendritic cell vaccine targeting Wilms’ Tumor 1 (WT1) antigen over the disease course (two points at MPE^1st and 2^nd, two months after MPE1^st). Epithelial cell adhesion molecule (EpCAM)⁺ cancer cells (PD-L1⁻ or T cell immunoglobulin mucin-3, TIM-3⁻), both PD-1 or TIM-3 positive CD8⁺ T cells, and CD14⁺CD68⁺CD163⁺TIM-3⁺ macrophages increased from the MPE^1st to MPE^2nd. The ratio of WT1-specific cytotoxic lymphocytes (WT1-CTLs) to MPE CD8⁺ T cells and IFN-γ secretion of WT1-CTLs were reduced with disease progression. Coincidentally, the fraction of central memory T (T_CM) of WT1-CTLs was decreased. On the other hand, CD8⁺ T cells in response to SMAD4^P130L, which is homogeneously expressed in EpCAM⁺ cancer cells, were detected using in vitro expansion with the HLA-A*11:01 restrictive SVCVNLYH neoantigen. Furthermore, the CD8⁺ T cell response to SMAD4^P130L was diminished following remarkably decreased numbers of CD8⁺ T_CM in MPE samples. In conclusion, CD8⁺ T cells responding to WT1 or SMAD4^P130L neoantigen expressed in EpCAM⁺ pancreatic cancer cells were detected in MPE. A tumor antigen-specific immune response would provide novel insight into the MPE microenvironment.     

Chitosan biobased and intelligent films: Monitoring pH variations

Many factors can influence the shelf-life product, such as pH, water activity, nutrient levels available oxygen, etc. Variations in food pH could occur due to microorganism growth and chemical reactions that may impact flavour, consistency and shelf-life. The objective of this paper was to obtain and characterize the fast pH-colourimetric indicator device, applying a simple manufacturing technique, using food grade and biodegradable materials. The intelligent film was based on natural compounds as chitosan and anthocyanin (pH-colourimetric indicator). Chitosan intelligent films (C-ATH, 2.0 g/100 g) were obtained incorporating anthocyanin (1.0 g/100 g) in matrix films. Initially, a dark violet colour was observed in chitosan films dried. When immersed in different pH buffers, the colour range of the films varied from pink (in acid pH) to bluish-green (in neutral pH) and to violet (in basic pH). The water solubility and water vapour transmission rate of C-ATH were decreased to 60% and 48%, respectively, as compared to control film (CF, without indicator). The mechanical properties such as tensile strength and rigidity were maintained, and the elongation at break was reduced to 47% compared to CF. The advantages of this system were the simple manufacturing process, biodegradability and usage of natural and safe compounds.     

A novel method for determining vitamin B1 in a wide variety of foodstuffs with or without polyphenols

Certain foodstuffs exhibit matrix interference effects on the vitamin B₁ analysis prescribed in the official methods of the European Union, AOAC International, and Japan. In this study, we demonstrated that one of the problematic polyphenols in green tea or cocoa was tannin. For these matrices, thiamine was found to interact with tannin in the extraction step and was thus removed by filtration as a residue. To resolve the matrix interference, we proposed lowering the pH of the extraction solution by adding hydrochloric acid after the enzyme reaction. High precision (2–3% RSDr) and good recovery (98.3–103%) were obtained with reference materials using the proposed method. We also confirmed the equivalence of the obtained data from the proposed method and the Japanese official method for nutritional labeling. From these results, the method was found to be effective for vitamin B₁ analysis regardless of the presence of interference matrices.     

Inhibitory effect of zinc on beta-hexosaminidase release from RBL-2H3 cells by synthetic chemicals

The effects of foods and chemicals related to food hygiene on degranulation were evaluated using a method for assaying the enzyme activity of beta-hexosaminidase as an index of chemical mediator release from RBL-2H3 cells in vitro. Using a previously developed assay system, we had found a large number of inhibitors and promoters of degranulation of RBL-2H3 cells. In the present study, we examined the inhibitory effect of zinc chloride on the degranulation (beta-hexosaminidase release) from RBL-2H3 cells with or without antigen in the presence of the degranulation-promotive chemicals, namely, 4 food additives, 7 pesticides and 2 veterinary drugs. These promotive chemicals were classified into two types on the basis of inhibitory profile by zinc chloride: 1) those which showed marked degranulation-inhibitory action when the cells were stimulated with antigen, such as butylhydroxyanisole, dibutylhydroxytoluene, EPN, cis- and trans-permethrin, prothiofos, pyridaben, terbufos, 2) those which showed marked degranulation-inhibitory action whether the cells were stimulated with antigen or not, such as butyl p-hydroxybenzoate, o-phenylphenol, bitertanol, salinomycin. In conclusion, zinc had a dramatic inhibitory effect on enhanced degranulation induced by synthetic chemicals in vitro.     

Subcellular localization of fructosyl amino acid oxidases in peroxisomes of Aspergillus terreus and Penicillium janthinellum

Fructosyl amino acid oxidase (FAOD) is the enzyme catalyzing the oxidative deglycation of Amadori compounds, such as fructosyl amino acids, yielding the corresponding amino acids, glucosone, and H(2)O(2). In a previous report, we determined the primary structures of cDNAs coding for FAODs from two fungal strains Aspergillus terreus AP1 and Penicillium janthinellum and we found that both fungal FAODs included the putative peroxisome targeting signal 1 (PTS1) at the carboxyl terminal (Yoshida, N. et al., Eur. J. Biochem., 242, 499-505, 1996). In this study, we determined the intracellular localization of FAODs in these two fungi. Subcellular fractionation experiments and immuno-electronmicroscopic observations, together with the previous findings indicated that the FAODs were localized in peroxisomes of A. terreus AP1 and P. janthinellum. These FAODs were also found to belong to a new member of "peroxisomal sarcosine oxidase family protein" in eucaryotic cells.     

Manipulation of micro solid particles and induced flows in water by laser irradiation

Q-switch for pulsing laser beam and Galvano scanner for rapid scanning laser beam were adapted to manipulatemicro solid particles in a water droplet.The trapping of fine particles at micron-sized diameters by a YAG lasersystem,induced flows,and the induced motion of the particles by laser beam irradiations are discussed.Particlerotations are observed by using anisotropic micro objects,and moreover the rotational rate with clockwise direc-tion is 22 r/min.The fragments of a cover glass were mixed in a water droplet,and their size and shape are un-even.The rotation of that non-spheres without scanning by the Galvano scanner is also observed in water.     

Localization of glycosaminoglycans and CD44 in the human lacrimal gland

Recent analyses of tears indicate the presence of glycosaminoglycans as their components, but their origin remains unknown. To further understand the origin of these tear components, we investigated by immunohistochemical techniques the localization of glycosaminoglycans and CD44 human lacrimal glands obtained from 20 cadavers at autopsy. Monoclonal antibodies to CD44, a receptor for hyaluronic acid, dermatan sulfate, chondroitin sulfate, and keratan sulfate were applied to the tissue. Hyaluronic acid binding region was also used for the staining of hyaluronic acid. By light microscopy, immunoreactivity for CD44 was mostly detected on the baso-lateral membrane of acinar and ductal cells, and the vascular endothelium in the interstitium. Positive staining of hyaluronic acid was associated intensely with the basal membrane of acinar and ductal cells and weakly, faintly or not at all with their lateral membrane. Positive staining of hyaluronic acid and immunoreactivity for dermatan sulfate were detected in interstitial fibrous structures; particularly, the former was intense in the perivascular fibrous structures, and the latter along the periparenchimal fibrous structures. Immunoreactivity for chondroitin sulfate and keratan sulfate was seen in some acinar cells and the acinar and ductal lumen. By electron microscopy, immunogold particles indication chondroitin sulfate sulfate or keratan sulfate labeled secretory granules of the acinar cells. Considering the fact that CD44 is a receptor molecule for hyaluronic acid, the association of hyaluronic acid with the basal membrane and weakly or faintly with the lateral membrane of acinar and ductal cells may be attributed to the expression of CD44 on the baso-lateral membrane of the cells. Moreover, the presence of immunoreactivity for chondroitin sulfate and keratan sulfate in secretory granules of acinar cells and their lumens suggests that tears from the lacrimal gland contain these glycosaminoglycans.