Effects of Repetitive Peripheral Magnetic Stimulation on Shoulder Subluxations Caused by Stroke: A Preliminary Study.

OBJECTIVES: Shoulder subluxation is a common problem after stroke. It causes shoulder pain that affects activities of daily living. This study aimed to investigate the effect of repetitive peripheral magnetic stimulation on shoulder subluxation after stroke. METHODS: We enrolled 12 consecutive patients who, as a result of stroke, suffered shoulder subluxations, measuring at half of a fingerbreadth or more. All subjects underwent conventional rehabilitation, as well as repetitive peripheral magnetic stimulation of their supraspinatus, posterior deltoid, and infraspinatus muscles. We assessed the following parameters: shoulder subluxation, evaluated as the acromio-humeral interval using measurements taken from X-rays; shoulder pain, evaluated using the Numerical Rating Scale; the active range of motion of shoulder abduction; and the motor impairment of the upper extremities, evaluated using the upper extremity of the Fugl-Meyer Assessment scale. RESULTS: The acromio-humeral interval before treatment was 22.8 ± 5.7 mm (mean ± SD). It significantly decreased to 19.6 ± 7.0 mm (p = 0.004) after treatment. Shoulder pain (p = 0.039), active range of motion of shoulder abduction (p = 0.016), and total (p = 0.005), subscale A (p = 0.005), and subscale C (p = 0.008) Fugl-Meyer Assessment scores also improved significantly after treatment. CONCLUSIONS: Repetitive peripheral magnetic stimulation effectively reduced shoulder subluxations and shoulder pain caused by stroke and improved voluntary upper-limb movements in stroke patients.

Repetitive Peripheral Magnetic Stimulation for Strengthening of the Suprahyoid Muscles: A Randomized Controlled Trial.

OBJECTIVE: Head lift exercise is a widely known form of training in the rehabilitation of patients with dysphagia. This study aimed to compare muscular strength reinforcement training of the suprahyoid muscles using repetitive peripheral magnetic stimulation (rPMS) with head lift exercises in a randomized controlled trial. MATERIALS AND METHODS: Twenty-four healthy adults were randomly assigned to either the magnetic stimulation group (M group) or the head lift exercise group (H group). Both groups underwent training five days a week for two weeks. The primary outcome was the cervical flexor strength, and secondary outcomes were jaw-opening force, tongue pressure, muscle fatigue of the hyoid and laryngeal muscles, displacement of the hyoid bone and opening width of the upper esophageal sphincter (UES) while swallowing 10 mL of liquid, training performance rate, and pain. RESULTS: No dropouts were reported during the two-week intervention period. Cervical flexor strength significantly increased solely in the M group. Tongue pressure significantly improved in both groups. There were no significant differences in the jaw-opening force, median frequency rate of the anterior belly of the digastric muscle, sternohyoid muscle, sternocleidomastoid muscle, anterior and superior hyoid bone displacement, and UES opening width in both groups. CONCLUSIONS: Two-week rPMS of the suprahyoid muscles increased the strength of these muscles compared with the head lift exercise during the same period.

Identification of cationic peptides derived from low protein rice by-products and evaluation of their multifunctional activities.

Low protein rice (LPR) by-products were used as a source of novel multifunctional cationic peptides. The LPR by-products were separated by ampholyte-free isoelectric focusing (autofocusing) into 20 fractions containing peptides with different isoelectric points (pIs). Subsequently, the antimicrobial activity of each fraction was evaluated against four pathogenic microorganisms. In addition, the cationic peptides from fractions exhibiting antimicrobial activity were purified using reversed-phase high-performance liquid chromatography and identified using matrix-assisted laser/desorption ionization-time-of-flight mass spectroscopy. Of the 11 cationic peptides identified, five peptides with pI values greater than 9.31 and net charges greater than +2 were chemically synthesized for multiple functionalities, including antimicrobial, lipopolysaccharide (LPS)-neutralizing, and angiogenic activities. Among these five cationic peptides, only LPR-KRK, which had a net charge of +9, exhibited antimicrobial activity against three of the four pathogenic microorganisms tested. Chromogenic LPS-neutralizing assays using Limulus amebocyte lysate showed that the 50% effective concentrations of these five peptides were between 0.11 and 3.09 µM. Tube-formation assays using human umbilical vein endothelial cells showed that all five peptides exhibited significant angiogenic activity at 1 µM and 10 µM, while none exhibited hemolytic activity toward mammalian red blood cells at concentrations up to 500 µM. Our results demonstrate that these five cationic peptides exhibit multiple biological functionalities with little or no hemolytic activity. Thus, fractions containing cationic peptides obtained from LPR by-products have the potential to be used as dietary supplements and functional ingredients in food products.

Hyoid Bone Movement at Rest by Peripheral Magnetic Stimulation of Suprahyoid Muscles in Normal Individuals.

OBJECTIVES: Neuromuscular electrical stimulation has been widely used in patients with dysphagia. However, obtaining sufficient hyoid bone movement through surface electrodes seems difficult. The aim of this study was to evaluate hyoid bone movement at rest through peripheral magnetic stimulation of the suprahyoid muscles in normal individuals. METHODS: Healthy adult men were recruited. A specially designed coil was connected to the peripheral magnetic stimulator. The coil was placed on the submental area of the subjects. Magnetic stimulation was performed at 30 Hz for 2 sec. The intensity level selected induces hyoid bone movement without causing intolerable pain to the subjects. The hyoid bone at rest between on- and off-magnetic stimulations of the suprahyoid muscles were identified using fluoroscopy at 30 frames/sec in lateral projection. Pain during peripheral magnetic stimulation was evaluated using the numerical rating scale (NRS). RESULTS: Eleven subjects aged 32 ± 9 years participated in this study. Magnetic stimulation resulted in 10.9 ± 2.8 mm forward displacement and 8.3 ± 4.1 mm (mean ± SD) upward displacement of the hyoid bone. The median NRS score during magnetic stimulation was 1. CONCLUSIONS: Peripheral magnetic stimulation is noninvasive and easy to perform. It does not require skin preparation, facilitates sufficient hyoid bone movement, and causes minimum level of pain.

Identification and characterization of multifunctional cationic peptides from traditional Japanese fermented soybean Natto extracts.

In this study, we investigated the lipopolysaccharide (LPS)-neutralizing and angiogenic activities of cationic peptides derived from the traditional Japanese fermented product Natto, which is made by fermenting cooked soybeans using Bacillus subtilis. Initially, we prepared 20 fractions of Natto extracts with various isoelectric points (pI's) using ampholyte-free isoelectric focusing (autofocusing). Cationic peptides were then purified from fractions 19 and 20, whose pH values were greater than 12, using reversed-phase high-performance liquid chromatography, and were identified using matrix-assisted laser/desorption ionization-time-of-flight mass spectroscopy. Among the 13 identified cationic peptides, seven (KFNKYGR, FPFPRPPHQK, GQSSRPQDRHQK, QRFDQRSPQ, ERQFPFPRPPHQK, GEIPRPRPRPQHPE, and EQPRPIPFPRPQPR) had pI's greater than 9.5, positive net charges, and differing molecular weights. These peptides were then chemically synthesized and applied to chromogenic LPS-neutralizing assays using Limulus amebocyte lysates, and 50% effective (neutralizing) concentrations of 2.6-5.5 µM were demonstrated. In addition, tube formation assays in human umbilical vein endothelial cells revealed angiogenic activities for all but one (GEIPRPRPRPQHPE) of these seven cationic peptides, with increases in relative tube lengths of 23-31% in the presence of peptides at 10 µM. Subsequent experiments showed negligible hemolytic activity of these peptides at concentrations of up to 500 µM in mammalian red blood cells. Collectively, these data demonstrate that six cationic peptides from Natto extracts, with the exception of GEIPRPRPRPQHPE, have LPS-neutralizing and angiogenic activities but do not induce hemolysis.

Clinical-oriented Three-dimensional Gait Analysis Method for Evaluating Gait Disorder.

Three-dimensional gait analysis (3DGA) is shown to be a useful clinical tool for the evaluation of gait abnormality due to movement disorders. However, the use of 3DGA in actual clinics remains uncommon. Possible reasons could include the time-consuming measurement process and difficulties in understanding measurement results, which are often presented using a large number of graphs. Here we present a clinician-friendly 3DGA method developed to facilitate the clinical use of 3DGA. This method consists of simplified preparation and measurement processes that can be performed in a short time period in clinical settings and intuitive results presentation to facilitate clinicians' understanding of results. The quick, simplified measurement procedure is achieved by the use of minimum markers and measurement of patients on a treadmill. To facilitate clinician understanding, results are presented in figures based on the clinicians' perspective. A Lissajous overview picture (LOP), which shows the trajectories of all markers from a holistic viewpoint, is used to facilitate intuitive understanding of gait patterns. Abnormal gait pattern indices, which are based on clinicians' perspectives in gait evaluation and standardized using the data of healthy subjects, are used to evaluate the extent of typical abnormal gait patterns in stroke patients. A graph depicting the analysis of the toe clearance strategy, which depicts how patients rely on normal and compensatory strategies to achieve toe clearance, is also presented. These methods could facilitate implementation of 3DGA in clinical settings and further encourage development of measurement strategies from the clinician's point of view.

Cationic peptides from peptic hydrolysates of rice endosperm protein exhibit antimicrobial, LPS-neutralizing, and angiogenic activities.

In this study, we hydrolyzed rice endosperm protein (REP) with pepsin and generated 20 fractions containing multifunctional cationic peptides with varying isoelectric point (pI) values using ampholyte-free isoelectric focusing (autofocusing). Subsequently, we determined antimicrobial activities of each fraction against the pathogens Prophyromonas gingivalis, Propionibacterium acnes, Streptocossus mutans, and Candida albicans. Fractions 18, 19, and 20 had pI values greater than 12 and exhibited antimicrobial activity against P. gingivalis, P. acnes, and C. albicans, but not against S. mutans. In further experiments, we purified and identified cationic peptides from fractions 18, 19, and 20 using reversed-phase high-performance liquid chromatography and matrix-assisted laser/desorption ionization-time-of-flight mass spectroscopy. We also chemically synthesized five identified peptides (RSVSKSR, RRVIEPR, ERFQPMFRRPG, RVRQNIDNPNRADTYNPRAG, and VVRRVIEPRGLL) with pI values greater than 10.5 and evaluated antimicrobial, lipopolysaccharide (LPS)-neutralizing, and angiogenic activities. Among these synthetic peptides, only VVRRVIEPRGLL exhibited antimicrobial activity against P. gingivalis, with an IC50 value of 87µM. However, all five cationic peptides exhibited LPS-neutralizing and angiogenic activities with little or no hemolytic activity against mammalian red blood cells at functional concentrations. These present data show dual or multiple functions of the five identified cationic peptides with little or no hemolytic activity. Therefore, fractions containing cationic peptides from REP hydrolysates have the potential to be used as dietary supplements and functional ingredients in food products.

Effect of visual biofeedback to acquire supraglottic swallow in healthy individuals: a randomized-controlled trial.

The aim of this study is to evaluate the effect of visual biofeedback therapy in acquiring supraglottic swallow (SGS) in a randomized-controlled trial with healthy individuals. Eighteen individuals (mean age, 26 years) who could not close or keep closed the vocal folds before and during the swallow in SGS were allocated randomly to either a visual biofeedback group (eight individuals) or a nonbiofeedback group (10 individuals). A videoendoscope was inserted intranasally and an SGS exercise, using 4 ml of green-colored water, was performed 30 times per day up to 5 days. When the participant failed to perform SGS, the result was provided only to the participants in the visual biofeedback group. The median length of time until acquiring SGS was 1.5 days in the visual biofeedback group and 3.5 days in the nonbiofeedback group (P=0.040). We concluded that visual biofeedback effectively enabled participants to acquire SGS earlier.

Endotoxin-neutralizing activity and mechanism of action of a cationic α-helical antimicrobial octadecapeptide derived from α-amylase of rice.

We have previously reported that AmyI-1-18, an octadecapeptide derived from α-amylase (AmyI-1) of rice, is a novel cationic α-helical peptide that exhibited antimicrobial activity against human pathogens, including Porphyromonas gingivalis, Pseudomonas aeruginosa, Propionibacterium acnes, Streptococcus mutans, and Candida albicans. In this study, to further investigate the potential functions of AmyI-1-18, we examined its inhibitory ability against the endotoxic activities of lipopolysaccharides (LPSs, smooth and Rc types) and lipid A from Escherichia coli. AmyI-1-18 inhibited the production of endotoxin-induced nitric oxide (NO), an inflammatory mediator, in mouse macrophages (RAW264) in a concentration-dependent manner. The results of a chromogenic Limulus amebocyte lysate assay illustrated that the ability [50% effective concentration (EC50): 0.17 µM] of AmyI-1-18 to neutralize lipid A was similar to its ability (EC50: 0.26 µM) to neutralize LPS, suggesting that AmyI-1-18 specifically binds to the lipid A moiety of LPS. Surface plasmon resonance analysis of the interaction between AmyI-1-18 and LPS or lipid A also suggested that AmyI-1-18 directly binds to the lipid A moiety of LPS because the dissociation constant (KD) of AmyI-1-18 with lipid A is 5.6×10(-10) M, which is similar to that (4.3×10(-10) M) of AmyI-1-18 with LPS. In addition, AmyI-1-18 could block the binding of LPS-binding protein to LPS, although its ability was less than that of polymyxin B. These results suggest that AmyI-1-18 expressing antimicrobial and endotoxin-neutralizing activities is useful as a safe and potent host defense peptide against pathogenic Gram-negative bacteria in many fields of healthcare.

Identification of proteinaceous inhibitors of a cysteine proteinase (an Arg-specific gingipain) from Porphyromonas gingivalis in rice grain, using targeted-proteomics approaches.

Porphyromonas gingivalis is known to be a major etiologic agent in the onset and progression of chronic periodontitis. Among various virulence factors that this bacterium produces, Arg- and Lys-specific cysteine proteinases (gingipains) are believed to be major determinants of the pathogenicity of P. gingivalis. Here, we report on our finding that there are inhibitors of these cysteine proteinases in a rice protein fraction. Comprehensive affinity chromatography and MS analyses resulted in the identification of 17 Arg-gingipain (Rgp)-interacting proteins in the rice endosperm. Of these, four proteins (i.e., a 26 kDa globulin, a plant lipid transfer/trypsin-alpha amylase inhibitor, the RA17 seed allergen, and an alpha amylase/trypsin inhibitor) were estimated to account for 90% of the Rgp inhibitory activity in the rice protein fraction, using a two-dimensional gel system of double-layer reverse zymography. In addition, a synthetic peptide derived from an Rgp-interacting protein, cyanate hydratase, could inhibit the growth of P. gingivalis and showed inhibitory activity against both the Arg- and Lys-gingipains. These results suggest that these rice proteins may be useful as nutraceutical ingredients for the prevention and management of periodontal diseases.

Functional Neuromuscular Stimulation for articular angle control with an Inverse Dynamics Model tuned by a neural network.

A Functional Neuromuscular Stimulation (FNS) system was developed to restore the motor function of people with central nervous system disorders. An FNS articular angle control system with an Inverse Dynamics Model (IDM) tuned by a neural network was proposed. The system was designed to control the elbow joint angles by learning the characteristics of the neuromuscular system of controlled limbs. Three kinds of control schemes using the IDM were proposed. In a simulation experiment, the system was able to learn the abnormal characteristics of a limb and any changes in these characteristics, and then allow for better control of the limb. The results of experiments using human participants showed that the system had ability to control both normal and paralysed limbs.

Ontogenetic changes in neuropeptide Y-immunoreactive cerebrospinal fluid-contacting neurons in the hypothalamus of the cloudy dogfish, Scyliorhinus torazame (Elasmobranchii).

Ontogenetic changes in neuropeptide Y-immunoreactive (NPY-ir) cerebrospinal fluid (CSF)-contacting neurons in the dogfish hypothalamus were studied immunohistochemically. NPY-ir CSF-contacting neurons first appeared in the median infundibular floor of the embryo at the 34 mm stage. At the 40 mm stage, similar neurons were found also in the saccus vasculosus (SV). The number of these neurons increased during the 54-80 mm stages, and the cells in the infundibular floor extended their basal processes to the neuropil of the median eminence, whereas the cells in the SV sent their axonal fibers to the tractus sacci vasculosi. After hatching, NPY immunoreactivity in the ventral hypothalamus became less dense, and the labeled CSF-contacting neurons tended to be confined to the nucleus lateralis tuberis, similarly as in the adults. The occurrence of NPY-ir CSF-contacting neurons in the SV was transient during the embryonic periods.